Sample records for egg extracts role

  1. Anthelmintic activity of acetone-water extracts against Haemonchus contortus eggs: interactions between tannins and other plant secondary compounds.

    PubMed

    Vargas-Magaña, J J; Torres-Acosta, J F J; Aguilar-Caballero, A J; Sandoval-Castro, C A; Hoste, H; Chan-Pérez, J A

    2014-12-15

    This study aimed at (i) describing the effects of acetone-water extracts obtained from a range of different plant materials, on the hatching process of Haemonchus contortus eggs under in vitro conditions and (ii) identifying the role of tannins and other plant secondary compounds (PSC), on these AH effects by using polyvinylpolypyrrolidone (PVPP), an inhibitor of tannins and other polyphenols. An egg hatch assay (EHA) was used to determine the AH effect. Acetone-water (70:30) extracts from different foliages (Lysiloma latisiliquum, Laguncularia racemosa, Rizophora mangle, Avicennia germinans) and plant by-products (Theobroma cacao seed husk and pulp, and percolated Coffea arabica) were obtained. Fresh H. contortus eggs were incubated in PBS with increasing concentrations of each extract (0, 600, 1200, 2400 and 3600 μg/ml PBS). A general linear model was used to determine the dose effect of each extract. A mild ovicidal activity was only recorded for T. cacao extracts (seed husk and pulp). The main anthelmintic (AH) effect for all the extracts, except for C. arabica, was to block the eclosion of larvated eggs. The use of PVPP at 3600 μg/ml PBS showed that tannins of the L. racemosa extract were responsible for blocking eclosion of larvated eggs. Extracts of L. latisiliquum, A. germinans, T. cacao seed husk and pulp also blocked eclosion of larvated eggs but the addition of PVPP indicated that tannins were not responsible for that activity. In contrast, it suggested unfavorable interactions between polyphenols and other PSC contained in those extracts, limiting the AH effect on the egg hatching process. The present results suggest that the interactions between tannins and other PSC are complex and may reduce the AH effects against H. contortus eggs. Copyright © 2014 Elsevier B.V. All rights reserved.

  2. Xenopus extract approaches to studying microtubule organization and signaling in cytokinesis

    PubMed Central

    Field, Christine M.; Pelletier, James F.; Mitchison, Timothy J.

    2017-01-01

    We report optimized methods for preparing actin-intact Xenopus egg extract. This extract is minimally perturbed, undiluted egg cytoplasm where the cell cycle can be experimentally controlled. It contains abundant organelles and glycogen, and supports active metabolism and cytoskeletal dynamics that closely mimic egg physiology. The concentration of the most abundant ~11,000 proteins is known from mass spectrometry. Actin-intact egg extract can be used for analysis of actin dynamics and interaction of actin with other cytoplasmic systems, as well as microtubule organization. It can be spread as thin layers, and naturally depletes oxygen though mitochondrial metabolism, which makes it ideal for fluorescence imaging. When combined with artificial lipid bilayers, it allows reconstitution and analysis of the spatially controlled signaling that positions the cleavage furrow during early cytokinesis. Actin-intact extract is generally useful for probing the biochemistry and biophysics of the large Xenopus egg. Protocols are provided for preparation of actin-intact egg extract, control of the cell cycle, fluorescent probes for cytoskeleton and cytoskeleton-dependent signaling, preparation of glass surfaces for imaging experiments, and immunodepletion to probe the role of specific proteins and protein complexes. We also describe methods for adding supported lipid bilayers to mimic the plasma membrane and for confining in microfluidic droplets to explore size scaling issues. PMID:28065319

  3. Embryotoxicity of an extract from Great Lakes lake trout to rainbow trout and lake trout

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Wright, P.J.; Tillitt, D.E.

    1995-12-31

    Aquatic ecosystems such as the Great Lakes are known to be contaminated with chemicals that are toxic to fish. However, the role of these contaminants in reproductive failures of fishes, such as lake trout recruitment, has remained controvertible. It was the objective to evaluate dioxin-like embryotoxicity of a complex mixture of chemicals and predict their potential to cause the lack of recruitment in Great Lakes lake trout. Graded doses of a complex environmental extract were injected into eggs of both rainbow trout and lake trout. The extract was obtained from whole adult lake trout collected from Lake Michigan in 1988.more » The extract was embryotoxic in rainbow trout, with LD50 values for Arlee strain and Erwin strain of 33 eggEQ and 14 eggEQ respectively. The LOAEL for hemorrhaging, yolk-sac edema, and craniofacial deformities in rainbow trout were 2, 2, and 4 eggEQ, respectively. Subsequent injections of the extract into lake trout eggs were likewise embryotoxic, with an LD50 value of 7 eggEQ. The LOAEL values for the extract in lake trout for hemorrhaging, yolk-sac edema, and craniofacial deformities were 0.1, 1, and 2 eggEQ, respectively. The current levels of contaminants in lake trout eggs are above the threshold for hemorrhaging and yolk-sac edema. The results also support the use of an additive model of toxicity to quantify PCDDs, PCDFs, Non-o-PCBs, and Mono-o-PCBs in relation to early life stage mortality in Lake Michigan lake trout.« less

  4. Xenopus egg extract: A powerful tool to study genome maintenance mechanisms.

    PubMed

    Hoogenboom, Wouter S; Klein Douwel, Daisy; Knipscheer, Puck

    2017-08-15

    DNA repair pathways are crucial to maintain the integrity of our genome and prevent genetic diseases such as cancer. There are many different types of DNA damage and specific DNA repair mechanisms have evolved to deal with these lesions. In addition to these repair pathways there is an extensive signaling network that regulates processes important for repair, such as cell cycle control and transcription. Despite extensive research, DNA damage repair and signaling are not fully understood. In vitro systems such as the Xenopus egg extract system, have played, and still play, an important role in deciphering the molecular details of these processes. Xenopus laevis egg extracts contain all factors required to efficiently perform DNA repair outside a cell, using mechanisms conserved in humans. These extracts have been used to study several genome maintenance pathways, including mismatch repair, non-homologous end joining, ICL repair, DNA damage checkpoint activation, and replication fork stability. Here we describe how the Xenopus egg extract system, in combination with specifically designed DNA templates, contributed to our detailed understanding of these pathways. Copyright © 2017. Published by Elsevier Inc.

  5. Analysis of signal transduction in cell-free extracts and rafts of Xenopus eggs.

    PubMed

    Tokmakov, Alexander A; Iwasaki, Tetsushi; Sato, Ken-Ichi; Fukami, Yasuo

    2010-05-01

    Intracellular signaling during egg activation/fertilization has been extensively studied using intact eggs, which can be manipulated by microinjection of different mRNAs, proteins, or chemical drugs. Furthermore, egg extracts, which retain high CSF activity (CSF-arrested extracts), were developed for studying fertilization/activation signal transduction, which have significant advantages as a model system. The addition of calcium to CSF-arrested extracts initiates a plethora of signaling events that take place during egg activation. Hence, the signaling downstream of calcium mobilization has been successfully studied in the egg extracts. Moreover, despite disruption of membrane-associated signaling compartments and ordered compartmentalization during extract preparation, CSF-arrested extracts can be successfully used to study early signaling events, which occur upstream of calcium release during egg activation/fertilization. In combination with the CSF-arrested extracts, activated egg rafts can reproduce some events of egg activation, including PLCgamma activation, IP3 production, transient calcium release, MAPK inactivation, and meiotic exit. This becomes possible due to complementation of the sperm-induced egg activation signaling machinery present in the rafts with the components of signal transduction system localized in the extracts. Herein, we describe protocols for studying molecular mechanisms of egg fertilization/activation using cell-free extracts and membrane rafts prepared from metaphase-arrested Xenopus eggs.

  6. The influence of volatile semiochemicals from stink bug eggs and oviposition-damaged plants on the foraging behaviour of the egg parasitoid Telenomus podisi.

    PubMed

    Michereff, M F F; Borges, M; Aquino, M F S; Laumann, R A; Mendes Gomes, A C M; Blassioli-Moraes, M C

    2016-10-01

    During host selection, physical and chemical stimuli provide important cues that modify search behaviours of natural enemies. We evaluated the influence of volatiles released by eggs and egg extracts of the stink bug Euschistus heros and by soybean plants treated with the eggs and egg extracts on Telenomus podisi foraging behaviour. Responses to volatiles were evaluated in Y-tube olfactometers after exposure to (1) one egg cluster for 24 h; (2) plants with eggs laid by the stink bug, tested at 24, 48, and 72 h after treatment; (3) plants with eggs laid artificially, tested at 24, 48, and 72 h after treatment; and (4) plants treated with acetone or hexane extracts of eggs. Telenomus podisi was attracted to volatiles emitted by one egg cluster and to acetone extracts of one egg cluster, but not to air or acetone controls. There were no responses to odours of plants treated with eggs or egg extracts. Analysis of acetone extracts of egg clusters by gas chromatography revealed the major components were saturated and unsaturated fatty acids, including hexadecanoic acid, linoleic acid, and (Z)-9-octadecenoic acid. Our results suggest that one egg cluster and the acetone extract of one egg cluster contain volatile compounds that can modify T. podisi foraging behaviour, and that the amounts of these compounds, probably together with some minor compounds, are important for host recognition by T. podisi. Also, the oviposition damage or egg extracts on the plant did not elicit indirect defences that attracted Telenomus podisi.

  7. Engineering Tough Materials: Biomimetic Eggshell

    DTIC Science & Technology

    2016-08-29

    known that this domain interacts with CO32- ions. Eggshell collagen membrane (Figure 1) was extracted by dissolving the shell of a chicken egg in 1 M...Therefore, an experiment was designed and executed with the aim of trying to elucidate whether the shell membrane has a mechanical role. Eggs 8 were...obtained from the local supermarket and holes were punctured at the equator in order to remove the liquid contents (both yolk and white). Eggs were tested in

  8. Biological effects of some natural and chemical compounds on the potato tuber moth, Phthorimaea operculella Zell. (Lepidoptera:Gelechiidae)

    PubMed Central

    Sharaby, Aziza; Abdel-Rahman, H.; Moawad, S.

    2009-01-01

    The olfactory reaction of larvae and moths was investigated towards 18 oils (6 natural oils and 12 commercial chemical oils). Some of these oils such as peppermint and camphor (natural oils) and eugenol and camphene (commercial oils) were repellent to both larvae and moths. Other oils such as strawberry and d-limonene were attractive to both larvae and moths. Some of the repellent oils were, therefore, tested for their effect on certain biological aspects of the insects. Eugenol and peppermint oils, each at the 0.01% conc., caused a significant depression in the fecundity of moth and decreased the percentage of egg hatchability. Eugenol oil was much more effective than peppermint oil at 1%. Dried (leaves, fruits or seeds) powder of 14 different plants species were tested in different concentrations with talcum powder (carrier material) against egg deposition. The results indicated that dried powders of Allium cepa, Curcuma longa, Colocasia antiqurum, Ocimum basilicum. Dodonaea viscose and Thuja orientalis played a highly significant role in reducing egg deposition. The most impressive effect was displayed by powders of D. viscose and A. cepa, which caused the highest depression in egg deposition as well as in the emerging offsprings. Ethanolic extracts of 11 plants indicated that extracts of Pithuranthos tortosus and Iphiona scabra caused the maximum inhibition of egg hatchability, followed by C. longa, Citrullus colocynthia and T. orientalis. Ethanolic extracts of Schinus terebenthiflius (leaves) and I. scabra caused the highest depression in the deposited eggs, as they played a remarkable role as ovipositor deterrents. The majority of the plant extracts at 1% conc. could protect potato tubers at different intervals according to the calculated tuber damage index as follows: Iphiopna > Pithuranthos > Curcuma > Schinus (fruits) Thuja > Schinus (leaves) > Dodonaea > Citrullus. PMID:23961036

  9. Biological effects of some natural and chemical compounds on the potato tuber moth, Phthorimaea operculella Zell. (Lepidoptera:Gelechiidae).

    PubMed

    Sharaby, Aziza; Abdel-Rahman, H; Moawad, S

    2009-07-01

    The olfactory reaction of larvae and moths was investigated towards 18 oils (6 natural oils and 12 commercial chemical oils). Some of these oils such as peppermint and camphor (natural oils) and eugenol and camphene (commercial oils) were repellent to both larvae and moths. Other oils such as strawberry and d-limonene were attractive to both larvae and moths. Some of the repellent oils were, therefore, tested for their effect on certain biological aspects of the insects. Eugenol and peppermint oils, each at the 0.01% conc., caused a significant depression in the fecundity of moth and decreased the percentage of egg hatchability. Eugenol oil was much more effective than peppermint oil at 1%. Dried (leaves, fruits or seeds) powder of 14 different plants species were tested in different concentrations with talcum powder (carrier material) against egg deposition. The results indicated that dried powders of Allium cepa, Curcuma longa, Colocasia antiqurum, Ocimum basilicum. Dodonaea viscose and Thuja orientalis played a highly significant role in reducing egg deposition. The most impressive effect was displayed by powders of D. viscose and A. cepa, which caused the highest depression in egg deposition as well as in the emerging offsprings. Ethanolic extracts of 11 plants indicated that extracts of Pithuranthos tortosus and Iphiona scabra caused the maximum inhibition of egg hatchability, followed by C. longa, Citrullus colocynthia and T. orientalis. Ethanolic extracts of Schinus terebenthiflius (leaves) and I. scabra caused the highest depression in the deposited eggs, as they played a remarkable role as ovipositor deterrents. The majority of the plant extracts at 1% conc. could protect potato tubers at different intervals according to the calculated tuber damage index as follows: Iphiopna > Pithuranthos > Curcuma > Schinus (fruits) Thuja > Schinus (leaves) > Dodonaea > Citrullus.

  10. The role of contact chemoreception in the host location process of an egg parasitoid.

    PubMed

    Iacovone, Alessia; French, Alice Sarah; Tellier, Frédérique; Cusumano, Antonino; Clément, Gilles; Gaertner, Cyril; Conti, Eric; Salerno, Gianandrea; Marion-Poll, Frédéric

    2016-01-01

    Taste allows insects to detect palatable or toxic foods, identify a mate, and select appropriate oviposition sites. The gustatory system strongly contributes to the survival and reproductive success of many species, yet it is rarely studied in insect parasitoids. In order to locate and assess a host in which they will lay their eggs, female wasps actively search for chemical cues using their sensory organs present mainly on the antennae. In this paper, we studied the role of antennal taste sensilla chaetica in the perception of contact semiochemicals in Trissolcus brochymenae (Hymenoptera: Platygastridae), an egg parasitoid of the brassicaceae pest Murgantia histrionica (Heteroptera: Pentatomidae). Methanolic extracts obtained from male and female hosts elicited action potentials in taste neurons housed in antennal sensilla chaetica, indicating that these sensilla are involved in the perception of non volatile host kairomones. In behavioural assays, wasp females displayed an intense searching behaviour in open arenas treated with host extracts, thus confirming that these kairomones are soluble in polar solvents. We further investigated the extracts by Gas Chromatography-Mass Spectrometry (GC-MS) and found that they contain several compounds which are good candidates for these contact kairomones. This study contributes to better understanding contact chemoreception in egg parasitoids and identifying gustatory receptor neurons involved in the host location process. Copyright © 2016 Elsevier Ltd. All rights reserved.

  11. Are the reductions in nematode attack on plants treated with seaweed extracts the result of stimulation of the formaldehyde cycle?

    PubMed

    Jenkins, T; Blunden, G; Wu, Y; Hankins, S D; Gabrielsen, B O

    1998-01-01

    Soil application to the roots of tomato plants (Lycopersicon esculentum) of a commercially-available alkaline extract of the brown alga, Ascophyllum nodosum, resulted in a significant reduction in the number of second-stage juveniles of both Meloidogynejavanica and M. incognita invading the roots, compared to those of plants treated with water alone. Egg recovery from the seaweed extract treated plants was also significantly lower. The three major betaines found in the seaweed extract (gamma-aminobutyric acid betaine, delta-aminovaleric acid betaine and glycinebetaine) also led to significant reductions in both the nematode invasion profile and egg recovery when applied at concentrations equivalent to those present in the extract. This led to the conclusion that the betaines present in the seaweed extract play a major role in bringing about the observed effects. Treatment of Arabidopsis thaliana plants with seaweed extract also resulted in a significant decrease in the number of females of M. javanica which developed in the roots. Significant reductions in egg recovery were also achieved from plants treated with the seaweed extract and similar effects were produced with the betaines found in the seaweed extract. As the experiments were conducted under monoxenic conditions, it can be concluded that the results obtained with the application of either the seaweed extract or betaines are not dependent on microorganisms associated with the rhizosphere.

  12. Comparative ovicidal activity of Moringa oleifera leaf extracts on Fasciola gigantica eggs

    PubMed Central

    Hegazi, Ahmed G.; Megeed, Kadria N. Abdel; Hassan, Soad E.; Abdelaziz, M. M.; Toaleb, Nagwa I.; Shanawany, Eman E. El; Aboelsoued, Dina

    2018-01-01

    Background: Fasciolosis is an important zoonotic disease affecting the productive performance of farm animals in Egypt. Aim: The aim of the present study was comparing the ovicidal effect of different extracts as an alcoholic (Methanolic and Ethanolic) and aqueous Moringa oleifera leaf extracts on Fasciola gigantica non-embryonated and developed eggs. Materials and Methods: Tested concentrations of extracts ranged from 12.5 to 800 mg/ml. Nitroxynil was used as reference drug with a dose of 100 mg/ml. Results: M. oleifera alcoholic and aqueous extracts showed a concentration-dependent ovicidal effect on F. gigantica non-embryonated and developed eggs. Based on LC50 values, water extract showed the highest ovicidal activity since it registered the lowest values of 2.6 mg/ml on non-embryonated eggs. Non-embryonated eggs were more susceptible to aqueous extract than developed eggs. On the other hand, the developed eggs were more susceptible to ethanolic extract than non-embryonated eggs even the lowest LC50 (12.38 mg/ml). Conclusion: M. oleifera leaf extracts especially aqueous extract could be a promising step in the field of controlling fascioliasis. Further, in vivo studies are needed to enlighten the therapeutic potential of M. oleifera extracts in treating F. gigantica infection. PMID:29657406

  13. Role of replication protein A as sensor in activation of the S-phase checkpoint in Xenopus egg extracts

    PubMed Central

    Recolin, Bénédicte; Van Der Laan, Siem; Maiorano, Domenico

    2012-01-01

    Uncoupling between DNA polymerases and helicase activities at replication forks, induced by diverse DNA lesions or replication inhibitors, generate long stretches of primed single-stranded DNA that is implicated in activation of the S-phase checkpoint. It is currently unclear whether nucleation of the essential replication factor RPA onto this substrate stimulates the ATR-dependent checkpoint response independently of its role in DNA synthesis. Using Xenopus egg extracts to investigate the role of RPA recruitment at uncoupled forks in checkpoint activation we have surprisingly found that in conditions in which DNA synthesis occurs, RPA accumulation at forks stalled by either replication stress or UV irradiation is dispensable for Chk1 phosphorylation. In contrast, when both replication fork uncoupling and RPA hyperloading are suppressed, Chk1 phosphorylation is inhibited. Moreover, we show that extracts containing reduced levels of RPA accumulate ssDNA and induce spontaneous, caffeine-sensitive, Chk1 phosphorylation in S-phase. These results strongly suggest that disturbance of enzymatic activities of replication forks, rather than RPA hyperloading at stalled forks, is a critical determinant of ATR activation. PMID:22187152

  14. Organochlorine contaminants in double-crested cormorants from Green Bay, WI: I. Large-scale extraction and isolation from eggs using semi-permeable membrane dialysis

    USGS Publications Warehouse

    Meadows, J.C.; Tillitt, D.E.; Schwartz, T.R.; Schroeder, D.J.; Echols, K.R.; Gale, R.W.; Powell, D.C.; Bursian, S.J.

    1996-01-01

    A 41.3-kg sample of double-crested cormorant (Phalacrocorax auritus) egg contents was extracted, yielding over 2 L of egg lipid. The double-crested cormorant (DCC) egg extract, after clean-up and concentration, was intended for use in egg injection studies to determine the embryotoxicity of the organic contaminants found within the eggs. Large-scale dialysis was used as a preliminary treatment to separate the extracted contaminants from the co-extracted sample lipids. The lipid was dialyzed in 80×5 cm semi-permeable membrane devices (SPMDs) in 50-ml aliquants. After the removal of 87 g of cholesterol by freeze-fractionation, the remaining lipid carryover (56 g) was removed by 100 routine gel permeation chromatography (GPC) operations. A 41,293-g sample was thus extracted and purified to the extent that it could easily be placed at a volume of 5 ml, the volume calculated to be necessary for the egg injection study. Analyses were performed comparing contaminant concentrations in the final purified extract to those present in the original egg material, in the extract after dialysis and cholesterol removal, and in the excluded materials. Recoveries of organochlorine pesticides through dialysis and cholesterol ranged from 96% to 135%. Total polychlorinated biphenyls in the final extract were 96% of those measured in the original egg material. Analysis of excluded lipid and cholesterol indicated that 92% of the polychlorinated dibenzo-dioxins and-furans were separated into the final extract.

  15. Skin testing with raw egg does not predict tolerance to baked egg in egg-allergic children.

    PubMed

    Turner, P J; Kumar, K; Fox, A T

    2014-11-01

    Most children with egg allergy tolerate egg in baked foods, such as cake, but tolerance cannot be predicted with conventional allergy testing. We hypothesized that the skin prick test (SPT) wheal to unprocessed raw egg might predict tolerance of baked egg at formal oral food challenge (OFC). We conducted a retrospective chart review to assess the utility of SPT wheal to egg extract (EE), raw egg (RE), and the ratio of EE:RE in predicting outcome of baked-egg OFC in children presenting to our tertiary referral centers with a physician diagnosis of egg allergy and following complete egg avoidance in their diet, between 2009 and 2013. OFC were performed following a standardized protocol using baked egg in cake, to a total dose equivalent to 3g egg protein. Data were analyzed from 186 completed challenges: OFC was positive in 64 (34%) children and negative in 122 (66%). Six children experienced anaphylaxis at OFC. Children tolerant to baked egg were more likely to have a lower SPT to egg extract/raw egg and EE:RE (median 0.56) than their allergic counterparts (0.70, p < 0.05). However, ROC curve analysis demonstrated poor predictivity of challenge outcome, with AUC for SPT to egg extract, raw egg and EE:ER equal to 0.71, 0.63 and 0.60, respectively. EE:RE was not helpful in predicting outcome of baked-egg OFC. Indeed, SPT to egg extract was slightly better at predicting outcome than either SPT to raw egg or EE:RE. Unfortunately, tolerance to baked egg can only be predicted from previous history or through controlled exposure. © 2014 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  16. Ecotoxicological assessment of diamondback terrapin (Malaclemys terrapin) pond habitat, prey and eggs in Bermuda.

    PubMed

    Outerbridge, Mark E; O'Riordan, Ruth; Fort, Douglas J; Davenport, John

    2016-01-15

    Total petroleum hydrocarbons, PAH and various trace metal residues were extracted and analyzed from fresh whole diamondback terrapin (Malaclemys terrapin) eggs, whole brackish-water gastropods (terrapin prey) and benthic sediment from anchialine pond environments in Bermuda inhabited by terrapins. Gastropods and terrapin eggs showed higher concentrations of trace metals and organic contaminants than sediments. Conversely, PAHs were mostly found within the sediment and smaller amounts detected in gastropods and terrapin eggs. Results indicated that contaminants in prey were transferred to terrapin eggs, and that concentrations of several contaminants exceeded potentially toxic concentrations for aquatic vertebrates. Necropsy of unhatched eggs from nests that had yielded viable hatchlings showed significantly compromised embryonic development. Bermudian diamondback terrapins reside and feed in brackish wetland habitats characterized by widespread, multifactorial contamination. This study suggests that environmental contamination plays a role in the recorded low hatching success in terrapin eggs in Bermuda. Copyright © 2015 Elsevier Ltd. All rights reserved.

  17. EGG-HATCHING MECHANISM OF HUMAN LIVER FLUKE, OPISTHORCHIS VIVERRINI: A ROLE FOR LEUCINE AMINOPEPTIDASES FROM THE SNAIL HOST, BITHYNIA SIAMENSIS GONIOMPHALOS.

    PubMed

    Tesana, Smarn; Khampoosa, P; Piratae, S; Prasopdee, S; Sripanidkulchai, B

    2018-05-08

    The human liver fluke Opisthorchis viverrini (Platyhelminthes, Trematoda, Digenea) uses snails of the genus Bithynia as first intermediate host. Peculiarly among trematodes, the eggs of O. viverrini hatch within the digestive tract of its snail host. It remains uncertain whether hatching in this species is mediated through mechanical fracture of the eggshell or by digestion with specific digestive enzymes. This study aimed to characterize enzymes with specific inhibitor and factors involved in the hatching activity of O. viverrini eggs. For measuring egg hatching in vivo, 50 O. viverrini mature eggs were fed to individual Bithynia siamensis goniomphalos snail at various temperature conditions for 24 hrs. Ex vivo, mature eggs were incubated with crude snail extract and commercial leucine aminopeptidase (LAP). Egg-hatching of O. viverrini was temperature dependent, with optimal hatching occurring at 24-28 C, with a peak of hatching of 93.54% in vivo and 30.55% ex vivo occurring at these temperatures. Ex vivo hatching rates increased to 45.87% under anaerobic conditions at 28 C. Some 22.70% and 16.21% of heat-killed eggs also hatched within the snail digestive tract and snail extract, respectively, indicating that host molecules are involved in the hatching response. Most eggs hatch in the anterior regions of the digestive tract. Hatching was completely inhibited in the presence of bestatin, an inhibitor of leucine aminopeptidase (LAP), but not in the presence of phosphatase inhibitors. Bestatin inhibition of hatching was reversible. Finally, egg hatching could be induced by addition of a porcine LAP. The results indicate that this digenean utilizes both LAP of the snail host and movement of miracidia for hatching.

  18. Xenopus egg cytoplasm with intact actin.

    PubMed

    Field, Christine M; Nguyen, Phuong A; Ishihara, Keisuke; Groen, Aaron C; Mitchison, Timothy J

    2014-01-01

    We report optimized methods for preparing Xenopus egg extracts without cytochalasin D, that we term "actin-intact egg extract." These are undiluted egg cytoplasm that contains abundant organelles, and glycogen which supplies energy, and represents the least perturbed cell-free cytoplasm preparation we know of. We used this system to probe cell cycle regulation of actin and myosin-II dynamics (Field et al., 2011), and to reconstitute the large, interphase asters that organize early Xenopus embryos (Mitchison et al., 2012; Wühr, Tan, Parker, Detrich, & Mitchison, 2010). Actin-intact Xenopus egg extracts are useful for analysis of actin dynamics, and interaction of actin with other cytoplasmic systems, in a cell-free system that closely mimics egg physiology, and more generally for probing the biochemistry and biophysics of the egg, zygote, and early embryo. Detailed protocols are provided along with assays used to check cell cycle state and tips for handling and storing undiluted egg extracts. © 2014 Elsevier Inc. All rights reserved.

  19. [Identification of special quality eggs with NIR spectroscopy technology based on symbol entropy feature extraction method].

    PubMed

    Zhao, Yong; Hong, Wen-Xue

    2011-11-01

    Fast, nondestructive and accurate identification of special quality eggs is an urgent problem. The present paper proposed a new feature extraction method based on symbol entropy to identify near infrared spectroscopy of special quality eggs. The authors selected normal eggs, free range eggs, selenium-enriched eggs and zinc-enriched eggs as research objects and measured the near-infrared diffuse reflectance spectra in the range of 12 000-4 000 cm(-1). Raw spectra were symbolically represented with aggregation approximation algorithm and symbolic entropy was extracted as feature vector. An error-correcting output codes multiclass support vector machine classifier was designed to identify the spectrum. Symbolic entropy feature is robust when parameter changed and the highest recognition rate reaches up to 100%. The results show that the identification method of special quality eggs using near-infrared is feasible and the symbol entropy can be used as a new feature extraction method of near-infrared spectra.

  20. Differential nuclear remodeling of mammalian somatic cells by Xenopus laevis oocyte and egg cytoplasm

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Alberio, Ramiro; Johnson, Andrew D.; Stick, Reimer

    2005-07-01

    The mechanisms governing nuclear reprogramming have not been fully elucidated yet; however, recent studies show a universally conserved ability of both oocyte and egg components to reprogram gene expression in somatic cells. The activation of genes associated with pluripotency by oocyte/egg components may require the remodeling of nuclear structures, such that they can acquire the features of early embryos and pluripotent cells. Here, we report on the remodeling of the nuclear lamina of mammalian cells by Xenopus oocyte and egg extracts. Lamin A/C is removed from somatic cells incubated in oocyte and egg extracts in an active process that requiresmore » permeable nuclear pores. Removal of lamin A/C is specific, since B-type lamins are not changed, and it is not dependent on the incorporation Xenopus egg specific lamin III. Moreover, transcriptional activity is differentially regulated in somatic cells incubated in the extracts. Pol I and II transcriptions are maintained in cells in oocyte extracts; however, both activities are abolished in egg extracts. Our study shows that components of oocyte and egg extracts can modify the nuclear lamina of somatic cells and that this nuclear remodeling induces a structural change in the nucleus which may have implications for transcriptional activity. These experiments suggest that modifications in the nuclear lamina structure by the removal of somatic proteins and the incorporation of oocyte/egg components may contribute to the reprogramming of somatic cell nuclei and may define a characteristic configuration of pluripotent cells.« less

  1. DNA extraction and barcode identification of development stages of forensically important flies in the Czech Republic.

    PubMed

    Olekšáková, Tereza; Žurovcová, Martina; Klimešová, Vanda; Barták, Miroslav; Šuláková, Hana

    2018-04-01

    Several methods of DNA extraction, coupled with 'DNA barcoding' species identification, were compared using specimens from early developmental stages of forensically important flies from the Calliphoridae and Sarcophagidae families. DNA was extracted at three immature stages - eggs, the first instar larvae, and empty pupal cases (puparia) - using four different extraction methods, namely, one simple 'homemade' extraction buffer protocol and three commercial kits. The extraction conditions, including the amount of proteinase K and incubation times, were optimized. The simple extraction buffer method was successful for half of the eggs and for the first instar larval samples. The DNA Lego Kit and DEP-25 DNA Extraction Kit were useful for DNA extractions from the first instar larvae samples, and the DNA Lego Kit was also successful regarding the extraction from eggs. The QIAamp DNA mini kit was the most effective; the extraction was successful with regard to all sample types - eggs, larvae, and pupari.

  2. Effects of lipid extraction on stable isotope ratios in avian egg yolk: Is arithmetic correction a reliable alternative?

    USGS Publications Warehouse

    Oppel, S.; Federer, R.N.; O'Brien, D. M.; Powell, A.N.; Hollmén, Tuula E.

    2010-01-01

    Many studies of nutrient allocation to egg production in birds use stable isotope ratios of egg yolk to identify the origin of nutrients. Dry egg yolk contains >50% lipids, which are known to be depleted in 13C. Currently, researchers remove lipids from egg yolk using a chemical lipid-extraction procedure before analyzing the isotopic composition of protein in egg yolk. We examined the effects of chemical lipid extraction on ??13C, ??15N, and ??34S of avian egg yolk and explored the utility of an arithmetic lipid correction model to adjust whole yolk ??13C for lipid content. We analyzed the dried yolk of 15 captive Spectacled Eider (Somateriafischeri) and 20 wild King Eider (S. spectabilis) eggs, both as whole yolk and after lipid extraction with a 2:1 chloroform:methanol solution. We found that chemical lipid extraction leads to an increase of (mean ?? SD) 3.3 ?? 1.1% in ??13C, 1.1 ?? 0.5% in ??15N, and 2.3 ?? 1.1% in ??34S. Arithmetic lipid correction provided accurate values for lipid-extracted S13C in captive Spectacled Eiders fed on a homogeneous high-quality diet. However, arithmetic lipid correction was unreliable for wild King Eiders, likely because of their differential incorporation of macronutrients from isotopically distinct environments during migration. For that reason, we caution against applying arithmetic lipid correction to the whole yolk ??13C of migratory birds, because these methods assume that all egg macronutrients are derived from the same dietary sources. ?? 2010 The American Ornithologists' Union.

  3. Ovicidal effect of the methanolic extract of ginger (Zingiber officinale) on Fasciola hepatica eggs: an in vitro study.

    PubMed

    Moazeni, Mohammad; Khademolhoseini, Ali Asghar

    2016-09-01

    Fasciolosis is of considerable economic and public health importance worldwide. Little information is available on the ovicidal effects of anthelminthic drugs. The use of ovicidal anthelmintics can be effective in disease control. In this study, the effectiveness of the methanolic extract of ginger (Zingiber officinale) on the eggs of Fasciola hepatica is investigated. Fasciola hepatica eggs were obtained from the gall bladders of naturally infected sheep and kept at 4 °C until use. The eggs were exposed to varying concentrations of ginger extract (1, 5, 10, 25 and 50 mg/mL) for 24, 48 and 72 h. To investigate the effect of the ginger extracts on the miracidial formation, the treated eggs were incubated at 28 °C for 14 days. The results indicated that F. hepatica eggs are susceptible to the methanolic extract of Z. officinale. The ovicidal effect of ginger extract at a concentration of 1 mg/mL with 24, 48 and 72 h treatment time was 46.08, 51.53 and 69.09 % respectively (compared with 22.70 % for control group). The ovicidal effect of ginger extract at a concentration of 5 mg/mL after 24 h was 98.84 %. One hundred percent ovicidal efficacy was obtained through application of ginger extract at concentrations of 5 and 10 mg/mL with a 48 and 24 h treatment time respectively. The in vitro ovicidal effect of the methanolic extract of Z. officinale was satisfactory in this study, however, in vivo efficacy of this extract, remains for further investigation. To the best of our knowledge, this is the first report on the ovicidal effect of Z. officinale against F. hepatica eggs.

  4. MCM interference during licensing of DNA replication in Xenopus egg extracts-Possible Role of a C-terminal region of MCM3.

    PubMed

    Mimura, Satoru; Kubota, Yumiko; Takisawa, Haruhiko

    2018-01-01

    The minichromosome maintenance (MCM) complex, consisting of six subunits, Mcm2-7, is loaded onto replication origins through loading factors (origin recognition complex [ORC], Cdc6, and Cdt1) and forms an MCM double hexamer that licenses the initiation of DNA replication. Previous studies with Xenopus egg extracts showed that loading factors, especially Cdc6, dissociate from chromatin on MCM loading, but the molecular mechanism and physiological significance remain largely unknown. Using a cell-free system for MCM loading onto plasmid DNA in Xenopus egg extracts, we found that MCM loaded onto DNA prevents DNA binding of the loading factors ORC, Cdc6, and Cdt1. We further report that a peptide of the C-terminal region of MCM3 (MCM3-C), previously implicated in the initial association with ORC/Cdc6 in budding yeast, prevents ORC/Cdc6/Cdt1 binding to DNA in the absence of MCM loading. ATP-γ-S suppresses inhibitory activities of both the MCM loaded onto DNA and the MCM3-C peptide. Other soluble factors in the extract, but neither MCM nor Cdt1, are required for the activity. Conservation of the amino acid sequences of MCM3-C and its activity in vertebrates implies a novel negative autoregulatory mechanism that interferes with MCM loading in the vicinity of licensed origins to ensure proper origin licensing.

  5. Effect of domestic cooking methods on egg yolk xanthophylls.

    PubMed

    Nimalaratne, Chamila; Lopes-Lutz, Daise; Schieber, Andreas; Wu, Jianping

    2012-12-26

    Xanthophylls are a class of bioactive compounds known to play an important role in preventing age-related macular degeneration. Egg yolk is a rich source of highly bioavailable xanthophylls including lutein and zeaxanthin. The effects of domestic cooking methods (boiling, frying, microwaving) on egg yolk xanthophyll content were investigated. A LC-(APCI)-MS/MS method was used to identify and quantify all-E- and Z-isomers of lutein, zeaxanthin, canthaxanthin, and β-apo-8'-carotenoic acid ethyl ester in fresh and cooked egg yolks. Both fresh and cooked yolks showed similar xanthophyll profiles but with higher contents of Z-isomers in cooked samples. All-E-lutein was the most affected, with 22.5%, 16.7%, and 19.3% reductions in boiled, microwaved, and fried yolk extracts, respectively. Total xanthophyll losses ranged from 6% to 18%. The results presented here could be useful in calculating the dietary intake of xanthophylls and also in assessing the xanthophyll profiles and contents of egg-containing products.

  6. DNA extraction in Echinococcus granulosus and Taenia spp. eggs in dogs stool samples applying thermal shock.

    PubMed

    Hidalgo, Alejandro; Melo, Angélica; Romero, Fernando; Hidalgo, Víctor; Villanueva, José; Fonseca-Salamanca, Flery

    2018-03-01

    The extraction of DNA in taeniid eggs shows complications attached to the composition of stool samples and the high resistance of eggs to degradation. The objective of this study was to test a method of DNA extraction in taeniid eggs by applying a thermal shock to facilitate the chemical-enzymatic degradation of these elements. A group of six tubes containing 1 ml of dog stool sample was spiked with eggs of Echinococcus granulosus and another group of six with Taenia pisiformis. Samples were floated with supersaturated sugar solution and centrifuged. The upper portion of each tube (500 μl) was aspirated and deposited in 1.5 ml tubes. Three tubes from each group were incubated at -20 °C and then at 90 °C, the remaining three from each group, incubated at room temperature. Proteinase K and lysis buffer were added to each tube and incubated for 12 h at 58 °C. The lysis effect was evaluated by microscopy at 3, 6 and 12 h and integrity by electrophoresis in 1% agarose gels. With the same experimental scheme, the thermal shock effect was evaluated in extractions of 1, 2, 3 and 4 eggs of each species and the DNA was quantified. Additionally, the protocol was applied in samples of 4 dogs diagnosed with natural infection by Taeniidae worms. Finally, all the extractions were tested by PCR amplification. Both E. granulosus and T. pisiformis eggs showed a similar response in the tests. In samples without treatment, the lysis effect was poor and showed no differences over time, but in those subjected to thermal shock, eggs degradation increased with time. In both treatments, there was no DNA loss integrity. The protocol applied to limited amounts of eggs yielded PCR products in 100% of the samples exposed to thermal shock, allowing PCR amplifications up to 1 egg. In non-exposed samples, the results were not replicable. However, DNA quantification showed low values in both treatments. In turn, DNA extractions with thermal shock in infected dog samples finally yielded PCR amplifications in 100%. It was concluded that thermal shock facilitates the DNA extraction for molecular analysis in taeniid eggs. The technique is effective extracting DNA even from a single egg and also to analyze natural infections samples with a relatively simple implementation. Published by Elsevier Inc.

  7. Preparation and use of Xenopus egg extracts to study DNA replication and chromatin associated proteins

    PubMed Central

    Gillespie, Peter J.; Gambus, Agnieszka; Blow, J. Julian

    2012-01-01

    The use of cell-free extracts prepared from eggs of the South African clawed toad, Xenopus laevis, has led to many important discoveries in cell cycle research. These egg extracts recapitulate the key nuclear transitions of the eukaryotic cell cycle in vitro under apparently the same controls that exist in vivo. DNA added to the extract is first assembled into a nucleus and is then efficiently replicated. Progression of the extract into mitosis then allows the separation of paired sister chromatids. The Xenopus cell-free system is therefore uniquely suited to the study of the mechanisms, dynamics and integration of cell cycle regulated processes at a biochemical level. In this article we describe methods currently in use in our laboratory for the preparation of Xenopus egg extracts and demembranated sperm nuclei for the study of DNA replication in vitro. We also detail how DNA replication can be quantified in this system. In addition, we describe methods for isolating chromatin and chromatin-bound protein complexes from egg extracts. These recently developed and revised techniques provide a practical starting point for investigating the function of proteins involved in DNA replication. PMID:22521908

  8. Rainbow trout embryotoxicity of a complex contaminant mixture extracted from Lake Michigan lake trout

    USGS Publications Warehouse

    Wilson, P.J.; Tillitt, D.E.

    1996-01-01

    Persistent Hydrophobic contaminants such as poly chlorinated dibenzo-p-dioxins, dibenzofurans and biphenyl congeners are present in aquatic systems, and are known to produce adverse effects in fish. Reproductive failure in fish populations has been observed in aquatic systems contaminated with persistent hydrophobic compounds. In order to mimic maternal transfer of environmental contaminants to newly fertilized fish eggs, a complex environmental extract was tested for embryotoxicity in a nanoinjection bioassay with embryos of rainbow trout. The extract was obtained from whole adult lake trout collected from Lake Michigan in 1988. The tissue extraction involved blending and dehydration with sodium sulfate, column extraction, dialysis separation, reactive cleanup and, finally, high-performance gel permeation chromatography. Egg gram-equivalent doses (g tissue/g egg normalized for egg % lipid) of the final extract (0.02, 0.10, 0.20, 1.0, 2.0, 4.0, 10.0, 20.0 eggEQ) were injected into newly fertilized rainbow trout eggs using triolein as the vehicle. The extract of the lake trout was embryotoxic to rainbow trout, with an LD50 of 35 eggEQ, based on total cumulative mortality. Gross physical abnormalities characteristic of dioxin exposure, such as hemorrhaging, yolk-sac edema and craniofacial deformities, were observed and showed significant dose-related increases. Sublethal effects in the rainbow trout, such as delayed time to hatch, mild hemorrhaging and moderate yolk-sac edema, resulted from estimated total PCB exposure as low as 8.8 ng/g, and this may have significant implications on Great Lakes lake trout fry and juvenile mortality.

  9. A simplified method for extracting androgens from avian egg yolks

    USGS Publications Warehouse

    Kozlowski, C.P.; Bauman, J.E.; Hahn, D.C.

    2009-01-01

    Female birds deposit significant amounts of steroid hormones into the yolks of their eggs. Studies have demonstrated that these hormones, particularly androgens, affect nestling growth and development. In order to measure androgen concentrations in avian egg yolks, most authors follow the extraction methods outlined by Schwabl (1993. Proc. Nat. Acad. Sci. USA 90:11446-11450). We describe a simplified method for extracting androgens from avian egg yolks. Our method, which has been validated through recovery and linearity experiments, consists of a single ethanol precipitation that produces substantially higher recoveries than those reported by Schwabl.

  10. Antioxidant effects of clove bud (Syzygium aromaticum) extract used with different extenders on ram spermatozoa during cryopreservation.

    PubMed

    Baghshahi, H; Riasi, A; Mahdavi, A H; Shirazi, A

    2014-12-01

    Clove bud (Syzygium aromaticum) extract was added at concentrations of 0, 35, 75, and 115 μg/ml to ovine semen extenders in order to investigate the antioxidant activities of clove bud extract and its effects on semen quality parameters after cryopreservation of ram spermatozoa. The basic extender was composed of Tris, egg yolk, and glycerol. Two other extenders were prepared by substitution of egg yolk with either LDL or egg yolk+SDS. The DPPH inhibition test was employed to assess the antioxidant activity of clove bud extract. Results showed that, compared to vitamin E, clove bud extract had a higher antioxidant activity. Better sperm motility and movement characteristics (P<0.05) were observed in the semen diluted with medium containing egg yolk+SDS than in that containing egg yolk and LDL. Progressive motility and movement characteristics of the sperm were significantly improved (P<0.05) by adding 35 and/or 75 μg/ml of clove bud extract to semen extenders. Sperm viability and plasma membrane integrity were also higher (P<0.05) in the semen exposed to medium containing egg yolk+SDS and 75 μg of clove buds extract after cryopreservation processes. Higher levels of clove bud extract, however, had adverse effects on all the sperm quality parameters and significantly reduced (P<0.05) the motility, movement parameters, viability, and plasma membrane integrity of ovine sperm. It was concluded that the clove bud extract had an antioxidant potential that makes it useful for addition to semen extenders and that the best results are obtained with a maximum clove bud extract of 75 μg/ml. Moreover, the combination of egg yolk and a detergent was found to improve sperm quality after the cooling and freeze-thawing processes. Copyright © 2014 Elsevier Inc. All rights reserved.

  11. Remodeling of ribosomal genes in somatic cells by Xenopus egg extract

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Ostrup, Olga, E-mail: osvarcova@gmail.com; Stem Cell Epigenetics Laboratory, Institute of Basic Medical Sciences, Faculty of Medicine, University of Oslo, Oslo; Norwegian Center for Stem Cell Research, Oslo

    Highlights: {yields} Xenopus egg extract remodels nuclei and alter cell growth characteristics. {yields} Ribosomal genes are reprogrammed within 6 h after extract exposure. {yields} rDNA reprogramming involves promoter targeting of SNF2H remodeling complex. {yields} Xenopus egg extract does not initiate stress-related response in somatic cells. {yields} Aza-cytidine elicits a stress-induced response in reprogrammed cells. -- Abstract: Extracts from Xenopus eggs can reprogram gene expression in somatic nuclei, however little is known about the earliest processes associated with the switch in the transcriptional program. We show here that an early reprogramming event is the remodeling of ribosomal chromatin and gene expression.more » This occurs within hours of extract treatment and is distinct from a stress response. Egg extract elicits remodeling of the nuclear envelope, chromatin and nucleolus. Nucleolar remodeling involves a rapid and stable decrease in ribosomal gene transcription, and promoter targeting of the nucleolar remodeling complex component SNF2H without affecting occupancy of the transcription factor UBF and the stress silencers SUV39H1 and SIRT1. During this process, nucleolar localization of UBF and SIRT1 is not altered. On contrary, azacytidine pre-treatment has an adverse effect on rDNA remodeling induced by extract and elicits a stress-type nuclear response. Thus, an early event of Xenopus egg extract-mediated nuclear reprogramming is the remodeling of ribosomal genes involving nucleolar remodeling complex. Condition-specific and rapid silencing of ribosomal genes may serve as a sensitive marker for evaluation of various reprogramming methods.« less

  12. Efficacy of Zingiber officinale ethanol extract on the viability, embryogenesis and infectivity of Toxocara canis eggs.

    PubMed

    El-Sayed, Nagwa Mostafa

    2017-12-01

    This study evaluated the effect of Zingiber officinal e ( Z. officinal e) ethanol extract on the viability, embryogenesis and infectivity Toxocara canis ( T. canis ) eggs. It was carried out both in vitro and in vivo. In the in vitro experiment, unembryonated T. canis eggs were incubated with 25, 50 and 100 mg/mL Z. officinal e extract at 25 °C for 6, 12, and 24 h to assess the effect of Z. officinal e on their viability and for two weeks to assess the effect of Z. officinal e on their embryogenesis. In vivo experiment was performed to assess the effect of Z. officinal e on infectivity of T. canis eggs. Treated embryonated eggs by Z. officinale extract at concentrations of 25, 50 and 100 mg/mL for 24 h were inoculated into mice and their livers were examined for the presence of T. canis larvae on the 7th day after infection and for histopathological evaluation at 14th day post-infection. Z. officinal e showed a significant ovicidal activity on T. canis eggs. The best effect was observed with 100 mg/mL concentration after 24 h with an efficacy of 98.2%. However, the treated eggs by 25, 50 mg/mL of Z. officinale extract after 24 h showed ovicidal activity by 59.22 and 82.5% respectively. Moreover, this extract effectively inhibited T. canis eggs embryogenesis by 99.64% and caused their degeneration at the concentration of 100 mg/mL after 2 weeks of treatment. However, the lower concentrations, 25 and 50 mg/mL inhibited embryogenesis by 51.19 and 78.57% respectively. The effect of Z. officinal e on the infectivity T. canis eggs was proven by the reduction of larvae recovery in the livers by 35.9, 62.8 and 89.5% in mice groups inoculated by Z. officinale treated eggs at concentrations of 25, 50 and 100 mg/mL respectively. Histopathologically, the liver tissues of mice infected with Z. officinale treated eggs at the concentration of 100 mg/mL appeared healthy with slight degenerative changes of hepatocytes, opposite to that recorded in the infected mice with treated eggs by the lower concentrations. In conclusion; Z. officinale extract possessed dose-dependent anti- T. canis activity on the viability, embryogenesis and infectivity of T. canis eggs.

  13. Identification of a novel C1q family member in color crucian carp (Carassius auratus) ovary.

    PubMed

    Chen, Bo; Gui, Jianfang

    2004-07-01

    Potential roles of C1q/tumor necrosis factor (TNF) superfamily proteins have been observed in vertebrate oogenesis and oocyte maturation, but no ovary-specific member has been identified so far. In this study, we have cloned and identified a novel member of C1q family with a C1q domain in the C-terminal from fully grown oocyte cDNA library of color crucian carp and demonstrated that the gene might be specifically expressed in ovary and therefore designated as Carassius auratus ovary-specific C1q-like factor, CaOC1q-like factor. It encodes a 213 amino acid protein with a 17 amino acid signal peptide. There is only one protein band of about 24.5 kDa in the extracts from phase I to phase IV oocytes, but two positive protein bands are detected in the extracts of mature eggs and fertilized eggs. Furthermore, the mobility shift of the smaller target protein band cannot be eliminated by phosphatase treatment, but the larger protein band increases its mobility on the gel after phosphatase treatment, suggesting that the larger protein might be a phosphorylated form. Immunofluorescence localization indicates that the CaOC1q-like proteins localize in cytoplasm, cytoplasm membrane and egg envelope of the oocytes at cortical granule stage and vitellogenesis stage, whereas they were compressed to cytoplasm margin in ovulated mature eggs and discharged into perivitelline space between cytoplasm membrane and egg envelope after egg fertilization. Further studies on distribution and translocation mechanism of the CaOC1q-like factor will be benefit to elucidate the unique function in oogenesis, oocyte maturation and egg fertilization.

  14. Combined extraction-cleanup column chromatographic procedure for determination of dicofol in avian eggs

    USGS Publications Warehouse

    Krynitsky, A.J.; Stafford, C.J.; Wiemeyer, Stanley N.

    1988-01-01

    Dicofol in avian eggs was completely oxidized to dichlorobenzophenone (DCBP) when a hexane Soxhlet extraction procedure was used. This degradation did not occur with other avian tissues (muscle and liver). For this reason, a combined extraction-cleanup column chromatographic procedure, without added heat, was developed for the determination of dicofol in avian eggs. Homogenized subsamples of eggs were mixed with sodium sulfate, and the mixture was added as the top layer on a column prepacked with Florisil. The dicofol and other compounds of interest were then eluted with ethyl ether-hexane. The extracts, relatively free from lipids, were quantitated on a gas chromatograph equipped with a 63Ni electron-capture detector and a methyl silicone capillary column. Recoveries from chicken eggs, fortified with dicofol and other DDT-related compounds, averaged 96%. Analysis of eggs of eastern screech-owls, fed a meat diet containing 10 ppm technical Kelthane, showed that both dicofol and DCBP were present. Results were confirmed by gas chromatography/mass spectrometry. This method is rapid and reliable, involves a minimum of sample handling, and is well suited for high volume determination of dicofol in eggs and other avian tissues.

  15. Experimental and field investigation of non-biting flies as potential mechanical vectors of Echinococcus granulosus eggs.

    PubMed

    Hemmati, Sahel; Afshar, Abbas Aghaei; Mohammadi, Mohammad Ali; Afgar, Ali; Nasibi, Saeid; Harandi, Majid Fasihi

    2018-06-01

    Synanthropic fly species can be potential mechanical vectors of many infectious agents. The potential of the flies to carry Echinococcus granulosus eggs is not fully documented. The purpose of the present study was to determine the possible role of non-biting flies to carry taeniid eggs. A total of 210 flies were collected from seven selected sites in areas of Kerman city, southeastern Iran from November 2016 to May 2017. Adult flies were live-caught using sweeping nets. Flies were placed individually in small glass bottles and transported to the laboratory. All the flies were killed by deep freezing and then identified to the species level using both morphological and molecular methods. The flies were homogenized in test tubes and genomic DNA was extracted and amplified by PCR. PCR protocols were used both to identify the live-caught flies to the species level, and for the detection of E. granulosus. The laboratory reared second generation flies were experimentally exposed to dog feces manually spiked by Echinococcus eggs. Two runs of experiments with 1-3 h of exposure were designed. For each experiment 20 flies were selected from the stock colony and were starved for three days. After each experiment, the flies were frozen for further molecular studies. The dominant fly species were Musca domestica and Lucilia sericata. No eggs were found on the body surface and/or guts of live-caught flies. After the first hour of exposure, 60%, of the flies of both species were found to harbor Echinococcus eggs. However, in the case of L. sericata 50% of the flies harbored Echinococcus eggs after 3 h of exposure. Results of the present study indicate the probable role of synanthropic flies in harboring Echinococcus eggs and mechanical transmission of cystic echinococcosis. When the helminth eggs are susceptible to desiccation grooming flies can remove many of eggs from exterior surfaces of them. Despite this result the role of synanthropic flies in the transmission of certain helminthiases should not be discounted because of their vagility and feeding mechanisms. Copyright © 2018 Elsevier Inc. All rights reserved.

  16. Antimitotic effect of an extract of the sea anemone Bunodosoma caissarum on sea urchin egg development.

    PubMed

    Malpezzi, E L; Freitas, J C

    1990-01-01

    A methanolic extract of the sea anemone Bunodosoma caissarum has an antimitotic effect on sea urchin egg development. The extract produces a dose-dependent inhibition of cell cleavage. When the extract is added together with sperm to unfertilized sea urchin eggs, the ED50 is 0.60 +/- 0.03 mg/ml (mean +/- SEM). When added shortly after fertilization, the extract produces the same kind of progressive inhibition but with an ED50 of 0.98 +/- 0.16 mg/ml. In the first case, detachment of the vitelline layer is inhibited whereas in the second case the extract inhibits cleavage even when the membrane is present.

  17. Large scale real-time PCR analysis of mRNA abundance in rainbow trout eggs in relationship with egg quality and post-ovulatory ageing.

    PubMed

    Aegerter, Sandrine; Jalabert, Bernard; Bobe, Julien

    2005-11-01

    The mRNA levels of 39 target genes were monitored in unfertilized eggs of 14 rainbow trout sampled the day of ovulation and again 5, 14, and 21 days later. For all 56 collected egg batches, an egg sample was fertilized to estimate egg quality by monitoring embryonic development. Remaining eggs were used for RNA extraction and subsequent real-time PCR analysis. A significant drop of egg quality was observed when eggs were held in the body cavity for 14 or 21 days post-ovulation (dpo). During the same period, eight transcripts (nucleoplasmin or Npm2, ferritin H, tubulin beta, JNK1, cyclin A1, cyclin A2, cathepsin Z, and IGF2) exhibited a differential abundance at one or several collection time(s). Interestingly, we observed higher levels of cyclins A1 and A2 mRNAs in eggs taken 5 days post-ovulation than in eggs taken, from the same females, at the time of ovulation. In addition, seven transcripts exhibited a differential abundance between low quality and high quality eggs. Low quality eggs were characterized by lower levels of Npm2, tubulin beta, and IGF1 transcripts. In contrast, keratins 8 and 18, cathepsin Z, and prostaglandin synthase 2 were more abundant in low quality eggs than in high quality eggs. In this study, we have demonstrated differences in mRNA levels in the rainbow trout egg that are reflective of developmental competence differences induced by post-ovulatory ageing. The putative role of these transcripts in post-ovulatory ageing-induced egg quality defects is discussed with special attention for corresponding cellular functions.

  18. In Vitro Anthelmintic Activity of Crude Extracts of Aerial Parts of Cissus quadrangularis L. and Leaves of Schinus molle L. against Haemonchus contortus.

    PubMed

    Zenebe, Selamawit; Feyera, Teka; Assefa, Solomon

    2017-01-01

    Haemonchus contortus, the causative agent of Haemonchosis, is the most economically important parasite in small ruminant production. Control with chemotherapy has not been successful due to rapid emergence of drug-resistant strains. There is a continuous search for alternative leads particularly from plants. The study aimed to evaluate the anthelmintic activity of crude methanolic extracts of leaves of Schinus molle and aerial parts of Cissus quadrangularis against H. contortus. Methods . Adult motility test and egg hatching inhibition assay were employed to investigate the in vitro adulticidal and egg hatching inhibitory effects of the extracts. Higher concentrations of the extracts (10 and 5 mg/ml) had a significantly superior adulticidal activity ( p < 0.05) compared to the negative control and lower concentration levels, which was comparable to albendazole. Similarly, the relative egg hatch inhibition efficacy of S. molle and C. quadrangularis extracts indicated a maximum of 96% and 88% egg hatch inhibition, respectively, within the 48 hrs of exposure at 1 mg/ml. The current study evidenced that the crude methanolic extracts of the plants have promising adulticidal and egg hatching inhibitory effects against H. contortus .

  19. Effect of Paecilomyces lilacinus, Trichoderma harzianum and Trichoderma virens fungal extracts on the hatchability of Ancylostoma eggs.

    PubMed

    Hofstätter, Bianca Delgado Menezes; Oliveira da Silva Fonseca, Anelise; de Souza Maia Filho, Fernando; de Souza Silveira, Julia; Persici, Beatriz Maroneze; Pötter, Luciana; Silveira, Andressa; Antoniolli, Zaida Inês; Brayer Pereira, Daniela Isabel

    Ancylostoma species have demanded attention due to their zoonotic potential. The use of anthelmintics is the usual method to prevent environmental contamination by Ancylostoma eggs and larvae. Nematophagous fungi have been widely used in their biological control due to the fungus ability to capture and digest free nematode forms. The aim of this study was to evaluate the effect of four different fungal extracts of Paecilomyces lilacinus (n=2), Trichoderma harzianum (n=1) and Trichoderma virens (n=1) isolates on the hatchability of Ancylostoma eggs. Fungal extracts consisted of fungal broth culture supernatant without filtration (crude extract) and filtered broth (filtered extract), macerated mycelium (crude macerate), and macerated mycelium submitted to filtration (filtered macerate). The Ancylostoma eggs were obtained from the feces of naturally infected dogs. In vitro assays were performed in five replicates and consisted of four treatments and one control group. The activity of the fungal extracts of each evaluated fungus differed (p<0.05) from those of the control group, showing significant ovicidal activity. The hatching of the eggs suffered reduction percentages of 68.43% and 47.05% with P. lilacinus, and 56.43% with T. harzianum, when crude macerate extract was used. The reduction with the macerate extract of T. virens was slightly lower (52.25%) than that for the filtered macerate (53.64%). The results showed that all extracts were effective in reducing the hatchability of Ancylostoma eggs. The ovicidal effect observed is likely to have been caused by the action of hydrolytic enzymes secreted by the fungi. Copyright © 2016 Asociación Española de Micología. Publicado por Elsevier España, S.L.U. All rights reserved.

  20. Organochlorine contaminants in double-crested cormorants from Green Bay, Wisconsin: II. Effects of an extract derived from cormorant eggs on the chicken embryo

    USGS Publications Warehouse

    Powell, D.C.; Aulerich, R.J.; Meadows, J.C.; Tillitt, D.E.; Stromborg, K.L.; Kubiak, T.J.; Giesy, J.P.; Bursian, S.J.

    1997-01-01

    White Leghorn chicken (Gallus domesticus) eggs were injected prior to incubation with one of four concentrations (0.001, 0.01, 0.1, and 1.0 egg-equivalent) of an extract derived from 1,000 double-crested cormorant (Phalacrocorax auritus) eggs collected at Spider Island adjacent to Green Bay in Lake Michigan. One egg-equivalent corresponded to the concentration of contaminants present in an average cormorant egg. This was approximately 322 pg toxic equivalents (TEQs)/g, ww egg with polychlorinatedbiphenyl congener 126 (3,3′,4,4′,5-pentachlorobiphenyl) accounting for over 70% of the TEQs. Injection of 1.0 egg-equivalent resulted in 77% mortality at hatch. The incidence of developmental abnormalities (structural defects or edema) was not affected by injection of the extract. Body weight gain of chicks was reduced in the 1.0 egg-equivalent dose group in the first, second, and third week's post-hatch. Relative brain weights were greater and relative bursa weights were less in the 1.0 egg-equivalent dose group than in the vehicle control at three weeks of age. There were no significant differences in the relative weights of the heart, liver, spleen, testes, or comb among treated and control birds.

  1. Histone H1 is essential for mitotic chromosome architecture and segregation in Xenopus laevis egg extracts

    PubMed Central

    Maresca, Thomas J.; Freedman, Benjamin S.; Heald, Rebecca

    2005-01-01

    During cell division, condensation and resolution of chromosome arms and the assembly of a functional kinetochore at the centromere of each sister chromatid are essential steps for accurate segregation of the genome by the mitotic spindle, yet the contribution of individual chromatin proteins to these processes is poorly understood. We have investigated the role of embryonic linker histone H1 during mitosis in Xenopus laevis egg extracts. Immunodepletion of histone H1 caused the assembly of aberrant elongated chromosomes that extended off the metaphase plate and outside the perimeter of the spindle. Although functional kinetochores assembled, aligned, and exhibited poleward movement, long and tangled chromosome arms could not be segregated in anaphase. Histone H1 depletion did not significantly affect the recruitment of known structural or functional chromosomal components such as condensins or chromokinesins, suggesting that the loss of H1 affects chromosome architecture directly. Thus, our results indicate that linker histone H1 plays an important role in the structure and function of vertebrate chromosomes in mitosis. PMID:15967810

  2. In Vitro Anthelmintic Activity of Crude Extracts of Aerial Parts of Cissus quadrangularis L. and Leaves of Schinus molle L. against Haemonchus contortus

    PubMed Central

    Zenebe, Selamawit; Feyera, Teka

    2017-01-01

    Background Haemonchus contortus, the causative agent of Haemonchosis, is the most economically important parasite in small ruminant production. Control with chemotherapy has not been successful due to rapid emergence of drug-resistant strains. There is a continuous search for alternative leads particularly from plants. The study aimed to evaluate the anthelmintic activity of crude methanolic extracts of leaves of Schinus molle and aerial parts of Cissus quadrangularis against H. contortus. Methods. Adult motility test and egg hatching inhibition assay were employed to investigate the in vitro adulticidal and egg hatching inhibitory effects of the extracts. Results Higher concentrations of the extracts (10 and 5 mg/ml) had a significantly superior adulticidal activity (p < 0.05) compared to the negative control and lower concentration levels, which was comparable to albendazole. Similarly, the relative egg hatch inhibition efficacy of S. molle and C. quadrangularis extracts indicated a maximum of 96% and 88% egg hatch inhibition, respectively, within the 48 hrs of exposure at 1 mg/ml. Conclusion The current study evidenced that the crude methanolic extracts of the plants have promising adulticidal and egg hatching inhibitory effects against H. contortus. PMID:29410960

  3. Ethanolic extracts of Inula viscosa, Salix alba and Quercus calliprinos, negatively affect the development of the entomopathogenic nematode, Heterorhabditis bacteriophora - A model to compare gastro-intestinal nematodes developmental effect.

    PubMed

    Santhi, Velayudhan Satheeja; Salame, Liora; Dvash, Levana; Muklada, Hussein; Azaizeh, Hassan; Mreny, Raghda; Awwad, Safaa; Markovics, Alex; Landau, Serge Yan; Glazer, Itamar

    2017-05-01

    Heterorhabditis bacteriophora can represent a model system for herbal medication against gastro-intestinal strongylid parasites in determining the recovery and development due to their unique parasitic infectious cycle. The fact that plant extracts impair nematode development is known but their differential impact on stages of the life cycle of H. bacteriophora has never been investigated. We examined the developmental stages resumed from eggs, young juveniles (J 1-3 ), infective juveniles (IJs), young and adult hermaphrodites of H. bacteriophora upon exposure to crude ethanolic extracts of Inula viscosa, Salix alba, and Quercus calliprinos at concentrations of 600, 1200, and 2400ppm. Our results showed that plant extracts were highly toxic to the survival of the eggs and young juveniles J 1 to J 3 at all concentrations. The plant extracts inhibited their development and were associated with low reproduction parameters (i.e. fecundity and viability of eggs). The IJs, J 4 , young and developed hermaphrodites displayed concentration-dependent negative effect on development with less egg count, poor vulval muscle development, loss of egg laying capacity and progeny development by matricidal hatching. Plant extract of I. viscosa at low (600ppm) concentration did not impair vulval development. These results suggest that these plant extracts show potential for the control of parasitic rhabditids. Copyright © 2017 Elsevier Inc. All rights reserved.

  4. IgY Technology: Extraction of Chicken Antibodies from Egg Yolk by Polyethylene Glycol (PEG) Precipitation

    PubMed Central

    Pauly, Diana; Chacana, Pablo A.; Calzado, Esteban G.; Brembs, Björn; Schade, Rüdiger

    2011-01-01

    Hens can be immunized by means of i.m. vaccination (Musculus pectoralis, left and right, injection volume 0.5-1.0 ml) or by means of Gene-Gun plasmid-immunization. Dependent on the immunogenicity of the antigen, high antibody-titres (up to 1:100,000 - 1:1,000,000) can be achieved after only one or 3 - 4 boost immunizations. Normally, a hen lays eggs continuously for about 72 weeks, thereafter the laying capacity decreases. This protocol describes the extraction of total IgY from egg yolk by means of a precipitation procedure (PEG. Polson et al. 1980). The method involves two important steps. The first one is the removal of lipids and the second is the precipitation of total IgY from the supernatant of step one. After dialysis against a buffer (normally PBS) the IgY-extract can be stored at -20°C for more than a year. The purity of the extract is around 80 %, the total IgY per egg varies from 40-80 mg, dependent on the age of the laying hen. The total IgY content increases with the age of the hen from around 40 mg/egg up to 80 mg/egg (concerning PEG precipitation). The laying capacity of a hen per year is around 325 eggs. That means a total potential harvest of 20 g total IgY/year based on a mean IgY content of 60 mg total IgY/egg (see Table 1). PMID:21559009

  5. Mutant analysis of Cdt1's function in suppressing nascent strand elongation during DNA replication in Xenopus egg extracts.

    PubMed

    Nakazaki, Yuta; Tsuyama, Takashi; Azuma, Yutaro; Takahashi, Mikiko; Tada, Shusuke

    2017-09-02

    The initiation of DNA replication is strictly regulated by multiple mechanisms to ensure precise duplication of chromosomes. In higher eukaryotes, activity of the Cdt1 protein is temporally regulated during the cell cycle, and deregulation of Cdt1 induces DNA re-replication. In previous studies, we showed that excess Cdt1 inhibits DNA replication by suppressing progression of replication forks in Xenopus egg extracts. Here, we investigated the functional regions of Cdt1 that are required for the inhibition of DNA replication. We constructed a series of N-terminally or C-terminally deleted mutants of Cdt1 and examined their inhibitory effects on DNA replication in Xenopus egg extracts. Our results showed that the region spanning amino acids (a. a.) 255-620 is required for efficient inhibition of DNA replication, and that, within this region, a. a. 255-289 have a critical role in inhibition. Moreover, one of the Cdt1 mutants, Cdt1 R285A, was compromised with respect to the licensing activity but still inhibited DNA replication. This result suggests that Cdt1 has an unforeseen function in the negative regulation of DNA replication, and that this function is located within a molecular region that is distinct from those required for the licensing activity. Copyright © 2017 Elsevier Inc. All rights reserved.

  6. MCM-BP regulates unloading of the MCM2–7 helicase in late S phase

    PubMed Central

    Nishiyama, Atsuya; Frappier, Lori; Méchali, Marcel

    2011-01-01

    Origins of DNA replication are licensed by recruiting MCM2–7 to assemble the prereplicative complex (pre-RC). How MCM2–7 is inactivated or removed from chromatin at the end of S phase is still unclear. Here, we show that MCM-BP can disassemble the MCM2–7 complex and might function as an unloader of MCM2–7 from chromatin. In Xenopus egg extracts, MCM-BP exists in a stable complex with MCM7, but is not associated with the MCM2–7 hexameric complex. MCM-BP accumulates in nuclei in late S phase, well after the loading of MCM2–7 onto chromatin. MCM-BP immunodepletion in Xenopus egg extracts inhibits replication-dependent MCM dissociation without affecting pre-RC formation and DNA replication. When excess MCM-BP is incubated with Xenopus egg extracts or immunopurified MCM2–7, it binds to MCM proteins and promotes disassembly of the MCM2–7 complex. Recombinant MCM-BP also releases MCM2–7 from isolated late-S-phase chromatin, but this activity is abolished when DNA replication is blocked. MCM-BP silencing in human cells also delays MCM dissociation in late S phase. We propose that MCM-BP plays a key role in the mechanism by which pre-RC is cleared from replicated DNA in vertebrate cells. PMID:21196493

  7. PIASy Mediates SUMO-2/3 Conjugation of Poly(ADP-ribose) Polymerase 1 (PARP1) on Mitotic Chromosomes*

    PubMed Central

    Ryu, Hyunju; Al-Ani, Gada; Deckert, Katelyn; Kirkpatrick, Donald; Gygi, Steven P.; Dasso, Mary; Azuma, Yoshiaki

    2010-01-01

    PIASy is a small ubiquitin-related modifier (SUMO) ligase that modifies chromosomal proteins in mitotic Xenopus egg extracts and plays an essential role in mitotic chromosome segregation. We have isolated a novel SUMO-2/3-modified mitotic chromosomal protein and identified it as poly(ADP-ribose) polymerase 1 (PARP1). PARP1 was robustly conjugated to SUMO-2/3 on mitotic chromosomes but not on interphase chromatin. PIASy promotes SUMOylation of PARP1 both in egg extracts and in vitro reconstituted SUMOylation assays. Through tandem mass spectrometry analysis of mitotically SUMOylated PARP1, we identified a residue within the BRCA1 C-terminal domain of PARP1 (lysine 482) as its primary SUMOylation site. Mutation of this residue significantly reduced PARP1 SUMOylation in egg extracts and enhanced the accumulation of species derived from modification of secondary lysine residues in assays using purified components. SUMOylation of PARP1 did not alter in vitro PARP1 enzyme activity, poly-ADP-ribosylation (PARylation), nor did inhibition of SUMOylation of PARP1 alter the accumulation of PARP1 on mitotic chromosomes, suggesting that SUMOylation regulates neither the intrinsic activity of PARP1 nor its localization. However, loss of SUMOylation increased PARP1-dependent PARylation on isolated chromosomes, indicating SUMOylation controls the capacity of PARP1 to modify other chromatin-associated proteins. PMID:20228053

  8. Effect of in ovo exposure to an organochlorine mixture extracted from double crested cormorant eggs (Phalacrocorax auritus) and PCB 126 on immune function of juvenile chickens

    USGS Publications Warehouse

    Lavoie, E.T.; Wiley, F.; Grasman, K.A.; Tillitt, D.E.; Sikarskie, J.G.; Bowerman, W.W.

    2007-01-01

    Organochlorine (OC) contaminants including polychlorinated biphenyls (PCBs) and p, p'-dichlorodiphenyldichloroethylene (DDE) have been associated with immune modulation in wild fish-eating birds from the Great Lakes. The objective of this study was to evaluate the immune function of juvenile chickens after in ovo exposure to PCB 126 or an environmentally relevant OC mixture extracted from eggs of double crested cormorants (Phalacrocorax auritus) from Green Bay, Lake Michigan, USA. Fertile white leghorn chicken (Gallus domesticus) eggs were injected before incubation with 0.55-1.79 ng TCDD equivalents (TEQ)/egg PCB 126 and 1.2-4.9 ng TEQs/egg of cormorant egg extract into the air cell in two separate experiments. After hatching, the immune function was tested using in vivo phytohemagglutinin (PHA) skin response in 11-day-old chicks, antibody titers to immunization with sheep red blood cells (SRBC) in 28-day-old chicks, and, at necropsy, thymus and bursal mass and cellularity. PCB 126 decreased antibody titers at all doses and decreased the thymus and bursa index but not cellularity at 1.79 ng TEQ/egg. The cormorant egg extract caused no significant alterations in immune function even though it has been demonstrated as immunotoxic in chicken embryos. However, twofold to threefold increases in total anti-SRBC titers in 28-day-old chicks exposed to 1.2 or 2.4 ng TEQ/egg of cormorant extract were similar to elevations in anti-SRBC titer observed in Caspian tern (Sterna caspia) chicks from a highly OC-contaminated site in Saginaw Bay, Lake Huron. Posthatch exposure to OC through fish consumption in addition to in ovo OC exposure might be associated with the immune modulation reported in wild birds. Chicks in this study might have begun to compensate for embryonic immunotoxicity by the ages at which we studied them. ?? 2007 Springer Science+Business Media, LLC.

  9. Signalling of Arabidopsis thaliana response to Pieris brassicae eggs shares similarities with PAMP-triggered immunity

    PubMed Central

    Reymond, Philippe

    2013-01-01

    Insect egg deposition activates plant defence, but very little is known about signalling events that control this response. In Arabidopsis thaliana, oviposition by Pieris brassicae triggers salicylic acid (SA) accumulation and induces the expression of defence genes. This is similar to the recognition of pathogen-associated molecular patterns (PAMPs), which are involved in PAMP-triggered immunity (PTI). Here, the involvement of known signalling components of PTI in response to oviposition was studied. Treatment with P. brassicae egg extract caused a rapid induction of early PAMP-responsive genes. In addition, expression of the defence gene PR-1 required EDS1, SID2, and, partially, NPR1, thus implicating the SA pathway downstream of egg recognition. PR-1 expression was triggered by a non-polar fraction of egg extract and by an oxidative burst modulated through the antagonistic action of EDS1 and NUDT7, but which did not depend on the NADPH oxidases RBOHD and RBOHF. Searching for receptors of egg-derived elicitors, a receptor-like kinase mutant, lecRK-I.8, was identified which shows a much reduced induction of PR-1 in response to egg extract treatment. These results demonstrate the importance of the SA pathway in response to egg-derived elicitor(s) and unravel intriguing similarities between the detection of insect eggs and PTI in Arabidopsis. PMID:23264520

  10. Determination of initiation of DNA replication before and after nuclear formation in Xenopus egg cell free extracts

    PubMed Central

    1993-01-01

    Xenopus egg extracts prepared before and after egg activation retain M- and S-phase specific activity, respectively. Staurosporine, a potent inhibitor of protein kinase, converted M-phase extracts into interphase- like extracts that were capable of forming nuclei upon the addition of sperm DNA. The nuclei formed in the staurosporine treated M-phase extract were incapable of replicating DNA, and they were unable to initiate replication upon the addition of S-phase extracts. Furthermore, replication was inhibited when the staurosporine-treated M- phase extract was added in excess to the staurosporine-treated S-phase extract before the addition of DNA. The membrane-depleted S-phase extract supported neither nuclear formation nor replication; however, preincubation of sperm DNA with these extracts allowed them to form replication-competent nuclei upon the addition of excess staurosporine- treated M-phase extract. These results demonstrate that positive factors in the S-phase extracts determined the initiation of DNA replication before nuclear formation, although these factors were unable to initiate replication after nuclear formation. PMID:8253833

  11. Modification of gDNA extraction from soil for PCR designed for the routine examination of soil samples contaminated with Toxocara spp. eggs.

    PubMed

    Borecka, A; Gawor, J

    2008-06-01

    A modification of gDNA extraction was developed for the polymerase chain reaction (PCR) technique, intended for the detection and differentiation of Toxocara spp. eggs in soil or sediments. Sand samples from sandpits confirmed as being contaminated with Toxocara spp. eggs by the flotation technique were analysed by PCR. The use of proteinase K made it possible to obtain genomic DNA from the sample without needing to isolate eggs using flotation or to inactivate PCR inhibitors present in the sand. Specific primers in the PCR reaction allowed discrimination between T. canis and T. cati eggs. The modification simplified the procedure, thanks to eliminating the step of gDNA isolation from eggs, which is both laborious and difficult.

  12. Effects of adding aqueous extract of Tribulus terrestris to diet on productive performance, egg quality characteristics, and blood biochemical parameters of laying hens reared under low ambient temperature (6.8 ± 3 °C)

    NASA Astrophysics Data System (ADS)

    Akbari, Mohsen; Torki, Mehran

    2016-06-01

    A study was conducted using 144 laying hens to evaluate the effects of adding aqueous extract of Tribulus terrestris to diets on productive performance, egg quality traits, and some blood parameters of laying hens reared under cold stress condition (6.8 ± 3 °C). The birds were randomly assigned to each of four dietary treatments (C, T1, T2, and T3) with six replicate cages of six birds. Diet inclusion of aqueous extract of T. terrestris at the rate of 10, 20, and 30 ml/Lit offered to groups T1, T2, and T3, respectively, while group C served as the control diet with no addition. Feed intake (FI), feed conversion ratio (FCR), egg weight (EW), egg production (EP), and egg mass (EM) were evaluated during the 42-day trial period. The EP and EM increased, whereas FCR decreased ( P < 0.001) in the hens fed the extract-included diet as compared to those fed the basal diet. The serum content of cholesterol decreased and the thickness of egg shell increased in the hens fed the T2 and T3 diet compared to those fed the basal diet. Overall from the results of the present experiment, it can be concluded that diet supplementation with aqueous extract of T. terrestris has beneficial effects on productive performance of laying hens reared under cold stress condition.

  13. Is it possible to make a diagnosis of raw, heated, and baked egg allergy in children using cutoffs? A systematic review.

    PubMed

    Calvani, Mauro; Arasi, Stefania; Bianchi, Annamaria; Caimmi, Davide; Cuomo, Barbara; Dondi, Arianna; Indirli, Giovanni Cosimo; La Grutta, Stefania; Panetta, Valentina; Verga, Maria Carmen

    2015-09-01

    The diagnosis of IgE-mediated egg allergy lies both on a compatible clinical history and on the results of skin prick tests (SPTs) and IgEs levels. Both tests have good sensitivity but low specificity. For this reason, oral food challenge (OFC) is the ultimate gold standard for the diagnosis. The aim of this study was to systematically review the literature in order to identify, analyze, and synthesize the predictive value of SPT and specific IgEs both to egg white and to main egg allergens and to review the cutoffs suggested in the literature. A total of 37 articles were included in this systematic review. Studies were grouped according to the degree of cooking of the egg used for OFC, age, and type of allergen used to perform the allergy workup. In children <2 years, raw egg allergy seems very likely when SPTs with egg white extract are ≥4 mm or specific IgEs are ≥1.7 kUA /l. In children ≥2 years, OFC could be avoided when SPTs with egg white extract are ≥10 mm or prick by prick with egg white is ≥14 mm or specific IgE is ≥7.3 kUA /l. Likewise, heated egg allergy can be diagnosed if SPTs with egg white extract are >5 and >11 mm in children <2 and ≥2 years, respectively. Further and better-designed studies are needed to determine the remaining diagnostic cutoff of specific IgE and SPT for heated and baked egg allergy. © 2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  14. Molluscicidal activities of aqueous extract of the sea anemone Parasicyonis actinostoloides against vector snails Bulinus truncatus and Lymnaea natalensis.

    PubMed

    el-Sayed, Kamelia Abass

    2006-08-01

    The aqueous extract of the sea anemone Parasicyonis actinostoloides showed molluscicidal effect against vector snails of Schistosoma hacematobium and Fasciola gigantica after 24 hours of exposure. LC50) and LC90 values for P. actinostoloides were 40 & 78.6 ppm for B. runcatus and 46.6 & 86.5 ppm for L. natalensis respectively. The effect of continuously exposure of B. truncatits and L. naltlensis to sublethal aqueous extract concentrations (LC0, LC10 & LC25) on survival rate, egg production and on infectivity of miracidia to infection with S. haematobium and F. gigantica were studied. The data showed that no B. truncatus survived more than 42, 32 & 27 days after exposure with a mean life span of 18.5, 13.3 & 11.1 days respectively. The death rate of B. truncatus with LC0 was highly significant as compared to treatment with LC10 & LC25 (p < 0.01). L. natalensis were more susceptible to the effect of aqueous extract than B. truncatus. LC0, LC10 & LC25, extract killed all L. natalensis through 32, 27 & 22 days. The mean life span of those exposed to LC0 was 12.37 days, high significant when compared with treated LC10 & LC25 ones (p < 0.01). The cumulative mortality rates of B. truncatus and L. natalensis in controls during the experimental study (52 days) was 60% & 75%, respectively. Egg production of B. truncatius and L. natalensis was not affected by sublethal concentrations. Control snails layed significantly higher no. of eggs than treated ones. B. truncatus stopped egg laying 17 days after exposure to LC25. those treated with LC10 & LC0 ceased to deposit eggs after 22 & 27 days respectively. The percent reduction in egg laying capacity of B. truncatus treated with LC0, LC10 & LC25 compared to controls was 77.1%, 93.2% & 92.8% respectively (p < 0.01). Similar reduction in egg production of treated L. natalensis cornpared to controls occurred, the percent reduction in egg production of snails treated with LC0, LC10 & LC25 in relation to controls was 78.4%, 92.4% & 94.7% respectively. Sublethal concentrations of aqueous extract of P. actinostoloides affected hatchability of B. truncatus and L. natalensis eggs. The data showed that eggs of B. truncatus and L. natalensis can hatch in all tested concentrations but with different rates. The eggs' hatchability in snails exposed to LC0, LCIo & LC25 extract at 5 days old was 44%, 38% & 30% in B. truncatus respectively. In L. natalensis eggs, the corresponding rates were lower 28%, 24% & 18% respectively. The infection of B. truncatuts and L. natalensis with S. haematobium n and F. giganlica miracidia was greatly reduced by the sublethal concentrations of aqueous extract of P. actinostoloides. The reduction of infection rate increased with the increased of sublethal concentrations. In B. truncatus the reduction was 43.2%, 57.6% & 76.6% compared to controls and in L. natalensis was 56.3%, 70.2% & 77.4%, respectively.

  15. Effects of processing on the recovery of food allergens from a model dark chocolate matrix.

    PubMed

    Khuda, Sefat E; Jackson, Lauren S; Fu, Tong-Jen; Williams, Kristina M

    2015-02-01

    To alleviate the risk to allergic consumers, it is crucial to improve factors affecting the detection of food allergens in processed chocolate products. This study evaluated processing effects on (1) recovery of peanut, egg, and milk allergens using five different extraction buffers, and (2) identification of specific allergenic proteins from extracts of incurred chocolate using allergen-specific antibodies and human allergic sera. Immunochemical staining with polyclonal antibodies showed that the addition of detergent or reducing agent improved extraction efficiency of peanut proteins, but not of egg and milk proteins. Tempering decreased antibody binding regardless of extractant. Detection of IgE-reactive peanut, egg, and milk allergens was differentially affected by tempering and extractant. Detection problems associated with matrix and processing effects may be overcome by the choice of extraction buffer and detecting antibody. Published by Elsevier Ltd.

  16. Vinegar decreases allergenic response in lentil and egg food allergy.

    PubMed

    Armentia, A; Dueñas-Laita, A; Pineda, F; Herrero, M; Martín, B

    2010-01-01

    Food allergy results from an atypical response of the mucosal immune system to orally consumed allergens. Antacid medication inhibits the digestion of dietary proteins and causes food allergy. A decrease of the gastric pH might enhance the function of digestion and reduce the risk of food allergy. To test a possible decrease in the allergenicity of powerful food allergens (egg, chicken, lentils) with the addition of vinegar during the cooking process. We included seven patients who suffered from anaphylaxis due to egg, chicken and lentils. We added vinegar to egg, chicken and lentil processed extracts used for skin prick tests (SPT) and compared the wheal areas obtained with the same extracts sources and the same way but without vinegar addition. Immunodetection was performed with the different processed extracts and patients' sera. Only one patient consented food challenge with vinegar-marinated-chicken. Wheal areas were significantly minor with the food extract with vinegar. Immunodetection showed a decrease of the response with vinegar processed extracts. Vinegar addition during the cooking process may decrease lentil and chicken allergenicity. Copyright 2009 SEICAP. Published by Elsevier Espana. All rights reserved.

  17. Intra-amniotic administration (Gallus gallus) of cicer arietinum and lens culinaris prebiotics extracts and duck egg white peptides affects calcium status and intestinal functionality

    USDA-ARS?s Scientific Manuscript database

    Calcium (Ca) is one of the most abundant inorganic elements in the human body and has many important physiological roles. Prebiotics and bioactive peptides are two important substances used to promote calcium uptake. However, the difference in mechanisms of the calcium uptake from these two suppleme...

  18. Dioxin-like toxic potency in Forster's tern eggs from Green Bay, Lake Michigan, North America

    USGS Publications Warehouse

    Tillitt, D. E.; Kubiak, T.J.; Ankley, G.T.; Giesy, J.P.

    1993-01-01

    The endangered Forster's tern (Sternaforsteri) population on Green Bay, Wisconsin has exhibited symptoms of embryotoxicity, congenital deformities, and poor hatching success. The putative causal agents are planar halogenated hydrocarbons (PHH). The objectives of this study were: 1) to evaluate the toxic potency of PHHs in extracts of Forster's tern eggs taken from Green Bay, Lake Michigan and a reference site, Lake Poygan, WI; and 2) to compare the toxic potencies of the egg extracts with the reproductive data available from the same water bird colonies. The relative toxic potency of the egg extracts was assessed with the H4IIE bioassay system to obtain 2,3,7,8-tetrachlorodibenzo-p-dioxin equivalents (TCDD-EQ). The average concentrations of TCDD-EQ in Forster's tern eggs were 214.5 pg/g and 23.4 pg/g from Green Bay and Lake Poygan, respectively. The bioassay results presented here concur with the biological effects and chemical analyses information from other studies on the same Forster's tern colonies.

  19. Effects of 3,3′,4,4′,5-pentachlorobiphenyl (PCB 126), 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), or an extract derived from field-collected cormorant eggs injected into double-crested cormorant (Phalacrocorax auritus) eggs

    USGS Publications Warehouse

    Powell, Debra C.; Aulerich, Richard J.; Meadows, John C.; Tillitt, Donald E.; Powell, Jon F.; Restum, Janelle C.; Stromborg, Kenneth L.; Giesy, John P.; Bursian, Steven J.

    1997-01-01

    Double-crested cormorant (Phalacrocorax auritus) eggs were injected with either 3,3′,4,4′,5-pentachlorobiphenyl (PCB 126), 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), or an extract derived from field-collected double-crested cormorant eggs. These compounds were injected into the yolks of cormorant eggs from an isolated colony on Lake Winnipegosis, Manitoba, Canada. Upon hatching, chicks were necropsied. The brain, bursa, heart, liver, and spleen were removed and weighed. An approximate median lethal dose (LD50) of 158 μg/kg egg was determined for PCB 126, which is 69 times greater than the LD50 determined for the chicken (Gallus domesticus) in a previous study. A significantly greater mortality occurred at the highest dose of TCDD (4.0 μg/kg egg) when compared to the vehicle control. However, the mortality data did not provide sufficient information for the determination of an LD50. The cormorant egg extract did not adversely affect hatchability. No significant increases were observed in the incidence of developmental abnormalities, including pronounced edema, in any of the treatment groups, nor were there any relevant effects on body and organ weights. Based on the results from this study, the cormorant appears to be considerably less sensitive to polyhalogenated diaromatic hydrocarbons than the chicken, which has been the typical species used for egg injection studies.

  20. Preparation of the egg membrane bandage contained the antibacterial Ag nanoparticles

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Zhang, Jin; Duan, Guangwen; Fu, Yunzhi, E-mail: yzhfu@hainu.edu.cn

    Silver nanoparticles were synthesized using a rapid, single step, and completely green biosynthetic method employing aqueous aloe leaf extracts as both the reducing and capping agent. Transmission electron microscopy analysis revealed the average size of silver nanoparticles approximately 18.05 nm. Fourier transform infrared spectroscopy observation showed the estimation of two kinds of binding sites between aqueous aloe leaf and aqueous aloe leaf with silver nanoparticles. In addition, the critical roles of the concentration of silver nitrate, temperature, and reaction time in the formation of silver nanoparticles had been illustrated. Furthermore, silver nanoparticles were deposited on egg membrane bandage, forming amore » new egg membrane bandage that contained silver nanoparticles that exhibiting excellent antibacterial effects against Gram-negative Escherichia coli and Gram-positive Staphylococcus aureus, which was 2.5 times stronger than the commercially available bandage. - Graphical Abstract: Display Omitted.« less

  1. Use of Moringa oleifera seed extracts to reduce helminth egg numbers and turbidity in irrigation water.

    PubMed

    Sengupta, Mita E; Keraita, Bernard; Olsen, Annette; Boateng, Osei K; Thamsborg, Stig M; Pálsdóttir, Guðný R; Dalsgaard, Anders

    2012-07-01

    Water from wastewater-polluted streams and dug-outs is the most commonly used water source for irrigation in urban farming in Ghana, but helminth parasite eggs in the water represent health risks when used for crop production. Conventional water treatment is expensive, requires advanced technology and often breaks down in less developed countries so low cost interventions are needed. Field and laboratory based trials were carried out in order to investigate the effect of the natural coagulant Moringa oleifera (MO) seed extracts in reducing helminh eggs and turbidity in irrigation water, turbid water, wastewater and tap water. In medium to high turbid water MO extracts were effective in reducing the number of helminth eggs by 94-99.5% to 1-2 eggs per litre and the turbidity to 7-11 NTU which is an 85-96% reduction. MO is readily available in many tropical countries and can be used by farmers to treat high turbid water for irrigation, however, additional improvements of water quality, e.g. by sand filtration, is suggested to meet the guideline value of ≤ 1 helminth egg per litre and a turbidity of ≤ 2 NTU as recommended by the World Health Organization and the U.S. Environmental Protection Agency for water intended for irrigation. A positive correlation was established between reduction in turbidity and helminth eggs in irrigation water, turbid water and wastewater treated with MO. This indicates that helminth eggs attach to suspended particles and/or flocs facilitated by MO in the water, and that turbidity and helminth eggs are reduced with the settling flocs. However, more experiments with water samples containing naturally occurring helminth eggs are needed to establish whether turbidity can be used as a proxy for helminth eggs. Copyright © 2012 Elsevier Ltd. All rights reserved.

  2. Ovicidal and larvicidal activity of crude extracts of Maesa lanceolata and Plectranthus punctatus against Haemonchus contortus.

    PubMed

    Tadesse, Dereje; Eguale, Tadesse; Giday, Mirutse; Mussa, Abiy

    2009-03-18

    The widespread development of anthelmintic resistance and high cost of the conventional anthelmintic drugs, has limited the control of gastrointestinal nematode parasites of sheep and goats and hence led to evaluation of medicinal plants as an alternative source of anthelmintics. In the current study, in vitro ovicidal and larvicidal activity of the leaves and fruits of the aqueous and hydro-alcoholic extracts of Maesa lanceolata and aerial parts of Plectranthus punctatus were evaluated on the egg and larvae of Haemonchus contortus using egg hatch assay and larval development test. All extracts of plants tested have shown complete inhibition of egg hatching at or below 1 mg/ml. ED50 for egg hatch inhibition ranged from 0.11 to 0.29 mg/ml, for both the aqueous and hydro-alcoholic extracts of Plectranthus punctatus and Maesa lanceolata. All extracts have shown dose dependent inhibition of larval development with variable results. The complete inhibition (100%) at the maximum concentration tested (50 mg/ml) was obtained only for hydro-alcoholic extract of the fruits of Maesa lanceolata and the lowest inhibition (50.33%) was recorded for the hydro-alcoholic extract of the leaves of the same plant. The overall findings of the present study has shown that Plectranthus punctatus and Maesa lanceolata contain possible anthelmintic compounds and further evaluation of different extracts and fractions of these plants should be carried out.

  3. Effects of adding aqueous extract of Tribulus terrestris to diet on productive performance, egg quality characteristics, and blood biochemical parameters of laying hens reared under low ambient temperature (6.8 ± 3 °C).

    PubMed

    Akbari, Mohsen; Torki, Mehran

    2016-06-01

    A study was conducted using 144 laying hens to evaluate the effects of adding aqueous extract of Tribulus terrestris to diets on productive performance, egg quality traits, and some blood parameters of laying hens reared under cold stress condition (6.8 ± 3 °C). The birds were randomly assigned to each of four dietary treatments (C, T1, T2, and T3) with six replicate cages of six birds. Diet inclusion of aqueous extract of T. terrestris at the rate of 10, 20, and 30 ml/Lit offered to groups T1, T2, and T3, respectively, while group C served as the control diet with no addition. Feed intake (FI), feed conversion ratio (FCR), egg weight (EW), egg production (EP), and egg mass (EM) were evaluated during the 42-day trial period. The EP and EM increased, whereas FCR decreased (P < 0.001) in the hens fed the extract-included diet as compared to those fed the basal diet. The serum content of cholesterol decreased and the thickness of egg shell increased in the hens fed the T2 and T3 diet compared to those fed the basal diet. Overall from the results of the present experiment, it can be concluded that diet supplementation with aqueous extract of T. terrestris has beneficial effects on productive performance of laying hens reared under cold stress condition.

  4. Antigenic cross-reactivity between Schistosoma mansoni and peanut: a role for cross-reactive carbohydrate determinants (CCDs) and implications for the hygiene hypothesis.

    PubMed

    Igetei, Joseph E; El-Faham, Marwa; Liddell, Susan; Doenhoff, Michael J

    2017-04-01

    The antigenic reactivity of constituents of Schistosoma mansoni and peanut (Arachis hypogaea) was investigated to determine whether identical antigenic epitopes possessed by both organisms provided a possible explanation for the negative correlation between chronic schistosome infection and atopy to allergens. Aqueous extracts of peanuts were probed in Western immunoblots with rabbit IgG antibodies raised against the egg, cercarial and adult worm stages of S. mansoni. Several molecules in the peanut extract were antigenically reactive with antibodies from the various rabbit anti-schistosome sera. A pair of cross-reactive peanut molecules at ~30 000-33 000 molecular weight was purified and both proteins were identified by mass spectrometric analysis as the peanut allergen Ara h 1. Anti-S. mansoni soluble egg antigen antibodies that were eluted off the peanut molecules reacted with two S. mansoni egg antigens identified by mass spectrometry as IPSE/α-1 and κ-5. Alignments of the amino acid sequences of Ara h 1 and either IPSE/α-1 or κ-5 revealed a low level of peptide sequence identity. Incubation of nitrocellulose paper carrying electrophoresed peanut molecules, six constituents of other allergic plants and S. mansoni egg antigens in a mild solution of sodium metaperiodate before probing with antibodies, inhibited most of the cross-reactivities. The results are consistent with the antigenic cross-reactive epitopes of S. mansoni egg antigens, peanut and other allergic plants being cross-reactive carbohydrate determinants (CCDs). These findings are novel and an explanation based on 'blocking antibodies' could provide an insight for the inverse relationship observed between schistosome infection and allergies. © 2017 John Wiley & Sons Ltd.

  5. Automated egg grading system using computer vision: Investigation on weight measure versus shape parameters

    NASA Astrophysics Data System (ADS)

    Nasir, Ahmad Fakhri Ab; Suhaila Sabarudin, Siti; Majeed, Anwar P. P. Abdul; Ghani, Ahmad Shahrizan Abdul

    2018-04-01

    Chicken egg is a source of food of high demand by humans. Human operators cannot work perfectly and continuously when conducting egg grading. Instead of an egg grading system using weight measure, an automatic system for egg grading using computer vision (using egg shape parameter) can be used to improve the productivity of egg grading. However, early hypothesis has indicated that more number of egg classes will change when using egg shape parameter compared with using weight measure. This paper presents the comparison of egg classification by the two above-mentioned methods. Firstly, 120 images of chicken eggs of various grades (A–D) produced in Malaysia are captured. Then, the egg images are processed using image pre-processing techniques, such as image cropping, smoothing and segmentation. Thereafter, eight egg shape features, including area, major axis length, minor axis length, volume, diameter and perimeter, are extracted. Lastly, feature selection (information gain ratio) and feature extraction (principal component analysis) are performed using k-nearest neighbour classifier in the classification process. Two methods, namely, supervised learning (using weight measure as graded by egg supplier) and unsupervised learning (using egg shape parameters as graded by ourselves), are conducted to execute the experiment. Clustering results reveal many changes in egg classes after performing shape-based grading. On average, the best recognition results using shape-based grading label is 94.16% while using weight-based label is 44.17%. As conclusion, automated egg grading system using computer vision is better by implementing shape-based features since it uses image meanwhile the weight parameter is more suitable by using weight grading system.

  6. Comparison of two extraction methods for the analysis of petroleum hydrocarbon residues in mallard duck eggs by GC and GC-MS

    USGS Publications Warehouse

    Belisle, A.A.; Gay, M.L.; Coon, N.C.

    1981-01-01

    Hydrocarbon residues in pooled eggs from a mallard duck on a diet of 25,000 ppm South Louisiana crude oil were compared after cleanup with and without saponification. The saponification procedure yielded superior reproducibility and extraction efficiency

  7. Comparison of two extraction methods for the analysis of petroleum hydrocarbon residues in mallard duck eggs by GC and GC-MS. [NONE

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Belisle, A.A.; Gay, M.L.; Coon, N.C.

    Hydrocarbon residues in pooled eggs from a mallard duck on a diet of 25,000 ppm South Louisiana crude oil were compared after cleanup with and without saponification. The saponification procedure yielded superior reproducibility and extraction efficiency..

  8. Omega-3 chicken egg detection system using a mobile-based image processing segmentation method

    NASA Astrophysics Data System (ADS)

    Nurhayati, Oky Dwi; Kurniawan Teguh, M.; Cintya Amalia, P.

    2017-02-01

    An Omega-3 chicken egg is a chicken egg produced through food engineering technology. It is produced by hen fed with high omega-3 fatty acids. So, it has fifteen times nutrient content of omega-3 higher than Leghorn's. Visually, its shell has the same shape and colour as Leghorn's. Each egg can be distinguished by breaking the egg's shell and testing the egg yolk's nutrient content in a laboratory. But, those methods were proven not effective and efficient. Observing this problem, the purpose of this research is to make an application to detect the type of omega-3 chicken egg by using a mobile-based computer vision. This application was built in OpenCV computer vision library to support Android Operating System. This experiment required some chicken egg images taken using an egg candling box. We used 60 omega-3 chicken and Leghorn eggs as samples. Then, using an Android smartphone, image acquisition of the egg was obtained. After that, we applied several steps using image processing methods such as Grab Cut, convert RGB image to eight bit grayscale, median filter, P-Tile segmentation, and morphology technique in this research. The next steps were feature extraction which was used to extract feature values via mean, variance, skewness, and kurtosis from each image. Finally, using digital image measurement, some chicken egg images were classified. The result showed that omega-3 chicken egg and Leghorn egg had different values. This system is able to provide accurate reading around of 91%.

  9. Effects of 3,3{prime},4,4{prime},5-pentachlorobiphenyl (PCB 126), 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), or an extract derived from field-collected cormorant eggs injected into double-crested cormorant (Phalacrocorax auritus) eggs

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Powell, D.C.; Aulerich, R.J.; Powell, J.F.

    1997-07-01

    Double-crested cormorant (Phalacrocorax auritus) eggs were injected with either 3,3{prime},4,4{prime},5-pentachlorobiphenyl (PCB 126), 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), or an extract derived from field-collected double-crested cormorant eggs. These compounds were injected into the yolks of cormorant eggs from an isolated colony on Lake Winnipegosis, Manitoba, Canada. Upon hatching, chicks were necropsied. The brain, bursa, heart, liver, and spleen were removed and weighed. An approximate median lethal dose (LD50) of 158 {micro}g/kg egg was determined for PCB 126, which is 69 times greater than the LD50 determined for the chicken (Gallus domesticus) in a previous study. A significantly greater mortality occurred at the highest dosemore » of TCDD when compared to the vehicle control. However, the mortality data did not provide sufficient information for the determination of an LD50. The cormorant egg extract did not adversely affect hatchability. No significant increases were observed in the incidence of developmental abnormalities, including pronounced edema, in any of the treatment groups, nor were there any relevant effects on body and organ weights. Based on the results from this study, the cormorant appears to be considerably less sensitive to polyhalogenated diaromatic hydrocarbons than the chicken, which has been the typical species used for egg injection studies.« less

  10. Plantago lanceolata and Plantago rugelii Extracts are Toxic to Meloidogyne incognita but not to Certain Microbes

    PubMed Central

    Meyer, Susan L. F.; Zasada, Inga A.; Roberts, Daniel P.; Vinyard, Bryan T.; Lakshman, Dilip K.; Lee, Jae-Kook; Chitwood, David J.; Carta, Lynn K.

    2006-01-01

    Extracts from the plants Plantago lanceolata and P. rugelii were evaluated for toxicity to the root-knot nematode Meloidogyne incognita, the beneficial microbes Enterobacter cloacae, Pseudomonas fluorescens and Trichoderma virens, and the plant-pathogenic fungi Fusarium oxysporum f. sp. gladioli, Phytophthora capsici, Pythium ultimum, and Rhizoctonia solani. Wild plants were collected, roots were excised from shoots, and the plant parts were dried and ground to a powder. One set of extracts (10% w/v) was prepared in water and another in methanol. Treatments included extract concentrations of 25%, 50%, 75% and 100%, and water controls. Meloidogyne incognita egg hatch was recorded after 7-day exposure to the treatments, and second-stage juvenile (J2) activity after 48 hours. All extracts were toxic to eggs and J2, with P. lanceolata shoot extract tending to have the most activity against M. incognita. Numbers of active J2 remained the same or decreased in a 24-hour water rinse following the 48-hour extract treatment, indicating that the extracts were lethal. When data from water- and methanol-extracted roots and shoots of both plant species were combined for analysis, J2 tended to be more sensitive than eggs to the toxic compounds at lower concentrations, while the higher concentrations (75% and 100%) were equally toxic to both life stages. The effective concentrations causing 50% reduction (EC50) in egg hatch and in J2 viability were 44.4% and 43.7%, respectively. No extract was toxic to any of the bacteria or fungi in our assays. PMID:19259537

  11. Catechol--an oviposition stimulant for cigarette beetle in roasted coffee beans.

    PubMed

    Nagasawa, Atsuhiko; Kamada, Yuji; Kosaka, Yuji; Arakida, Naohiro; Hori, Masatoshi

    2014-05-01

    The cigarette beetle, Lasioderma serricorne, is a serious global pest that preys on stored food products. Larvae of the beetle cannot grow on roasted coffee beans or dried black or green tea leaves, although they oviposit on such products. We investigated oviposition by the beetles on MeOH extracts of the above products. The number of eggs laid increased with an increase in dose of each extract, indicating that chemical factors stimulate oviposition by the beetles. This was especially true for \\ coffee bean extracts, which elicited high numbers of eggs even at a low dose (0.1 g bean equivalent/ml) compared to other extracts. Coffee beans were extracted in hexane, chloroform, 1-butanol, MeOH, and 20% MeOH in water. The number of eggs laid was higher on filter papers treated with chloroform, 1-butanol, MeOH, and 20% MeOH in water extracts than on control (solvent alone) papers. The chloroform extract was fractionated by silica-gel column chromatography. Nine compounds were identified by gas chromatography/mass spectrometry from an active fraction. Of these compounds, only a significant ovipositional response to catechol was observed.

  12. A Simple and Fast Extraction Method for the Determination of Multiclass Antibiotics in Eggs Using LC-MS/MS.

    PubMed

    Wang, Kun; Lin, Kunde; Huang, Xinwen; Chen, Meng

    2017-06-21

    The purpose of this study was to develop and validate a simple, fast, and specific extraction method for the analysis of 64 antibiotics from nine classes (including sulfonamides, quinolones, tetracyclines, macrolides, lincosamide, nitrofurans, β-lactams, nitromidazoles, and cloramphenicols) in chicken eggs. Briefly, egg samples were simply extracted with a mixture of acetonitrile-water (90:10, v/v) and 0.1 mol·L -1 Na 2 EDTA solution assisted with ultrasonic. The extract was centrifuged, condensed, and directly analyzed on a liquid chromatography coupled to tandem mass spectrometry. Compared with conventional cleanup methods (passing through solid phase extract cartridges), the established method demonstrated comparable efficiencies in eliminating matrix effects and higher or equivalent recoveries for most of the target compounds. Typical validation parameters including specificity, linearity, matrix effect, limits of detection (LODs) and quantification (LOQs), the decision limit, detection capability, trueness, and precision were evaluated. The recoveries of target compounds ranged from 70.8% to 116.1% at three spiking levels (5, 20, and 50 μg·kg -1 ), with relative standard deviations less than 14%. LODs and LOQs were in the ranges of 0.005-2.00 μg·kg -1 and 0.015-6.00 μg·kg -1 for all of the antibiotics, respectively. A total of five antibiotics were successfully detected in 22 commercial eggs from local markets. This work suggests that the method is suitable for the analysis of multiclass antibiotics in eggs.

  13. MTBP, the partner of Treslin, contains a novel DNA-binding domain that is essential for proper initiation of DNA replication.

    PubMed

    Kumagai, Akiko; Dunphy, William G

    2017-11-01

    Treslin, which is essential for incorporation of Cdc45 into the replicative helicase, possesses a partner called MTBP (Mdm2-binding protein). We have analyzed Xenopus and human MTBP to assess its role in DNA replication. Depletion of MTBP from Xenopus egg extracts, which also removes Treslin, abolishes DNA replication. These extracts be can rescued with recombinant Treslin-MTBP but not Treslin or MTBP alone. Thus, Treslin-MTBP is collectively necessary for replication. We have identified a C-terminal region of MTBP (the CTM domain) that binds efficiently to both double-stranded DNA and G-quadruplex (G4) DNA. This domain also exhibits homology with budding yeast Sld7. Mutants of MTBP without a functional CTM domain are defective for DNA replication in Xenopus egg extracts. These mutants display an impaired localization to chromatin and the inability to support loading of Cdc45. Human cells harboring such a mutant also display severe S-phase defects. Thus, the CTM domain of MTBP plays a critical role in localizing Treslin-MTBP to the replication apparatus for initiation. © 2017 Kumagai and Dunphy. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

  14. Effects of rhaponticum carthamoides versus glycyrrhiza glabra and punica granatum extracts on metabolic syndrome signs in rats

    PubMed Central

    2014-01-01

    Background Rhaponticum cathamoides (RC) is an endemic wild Siberian herb with marked medicinal properties that are still poorly understood. The aim of this study is to investigate the therapeutic potential of RC extract (ERC) compared to the effects of Glycyrrhiza glabra (EGG) and Punica granatum extracts (EPG) in a rat model with high-fat diet-(HFD)-induced signs of metabolic syndrome; therefore, this study addresses a significant global public health problem. Methods Six-month-old male Wistar Albino Glaxo rats were subjected to eight weeks of a standard diet (SD), HFD, or HFD in which ERC, EGG, or EPG powders were incorporated at 300 mg/kg/day. The serum lipid profile, corticosterone and cytokine concentrations, glucose tolerance, systolic blood pressure, triacylglycerol accumulation, and PPARα DNA-binding activities in the liver samples were determined. Results In contrast to EGG and EPG, an ERC supplement significantly reduced the weight of epididymal tissue (19.0%, p < 0.01) and basal serum glucose level (19.4%, p < 0.05). ERC improved glucose intolerance as well as dyslipidemia more efficiently than EGG and EPG. EGG but not ERC or EPG supplementation decreased systolic blood pressure by 12.0% (p < 0.05). All of the tested extracts reduced serum IL6 and corticosterone levels induced by HFD. However, the lowering effects of ERC consumption on the serum TNF-α level and its restoring effect on the adrenal corticosterone level significantly exceeded the improvements induced by EGG and EPG. ERC intake also reduced triacylglycerol accumulation and increased the PPARα DNA-binding activity in the liver more significantly than EGG and EPG. Conclusions ERC powder supplementation improved glucose and lipid metabolism more significantly than EGG and EPG in rats fed on HFD, supporting the strategy of R. carthamoides use for safe relief of metabolic syndrome and its related disturbances such as inflammation, stress, and hepatic steatosis. PMID:24444255

  15. Enzymatic modification of egg lecithin to improve properties.

    PubMed

    Asomaning, Justice; Curtis, Jonathan M

    2017-04-01

    This research studied the enzymatic modification of egg yolk phospholipids and its effect on physicochemical properties. Egg yolk lipids were extracted with food grade ethanol and egg phospholipids (ePL) produced by deoiling with acetone. Vegetable oils were used to interesterify ePL utilizing Lipozyme®: sn-1,3 specific lipase. The enzymatic interesterification resulted in a single phase liquid product, whereas simple blending of the ePL and vegetable oil resulted in a product with two phases. In addition solid fat content decreased by 50% at -10°C and 94% at 35°C when compared with egg yolk lipids extract. A decrease in melting temperature resulted from the interesterification process. Interesterification improved emulsion stability index when used as an emulsifier in oil-in-water emulsion and compared to the native and soy lecithin. Enzyme reusability test showed retention of 63% activity after 10 cycles. Overall, the properties of native egg phospholipids were significantly enhanced in a potentially useful manner through interesterification. Copyright © 2016 Elsevier Ltd. All rights reserved.

  16. Comparative study to evaluate the anti-viral efficacy of Glycyrrhiza glabra extract and ribavirin against the Newcastle disease virus

    PubMed Central

    Omer, Muhammad Ovais; AlMalki, Waleed Hassan; Shahid, Imran; Khuram, Shahzada; Altaf, Imran; Imran, Saeed

    2014-01-01

    Background: The Newcastle disease represents as one of the most infectious viral disease, which afflicts almost every species of the birds. The causative agent of the disease is a single-stranded RNA virus with rapid replication capability. Objective: This study was performed to evaluate the comparative anti-viral efficacy and toxicity of Glycyrrhiza glabra aqueous extract and ribavirin against the Newcastle disease virus. Materials and Methods: The embryonated eggs were divided into six groups (A, B, C, D, E and F). Groups A, B, C, and D were further subdivided into three subgroups. The virus was identified by hemagglutination inhibition test. Spot hemagglutination test and viability of embryos were also evaluated. Three different concentrations i-e., 30 mg/100 ml, 60 mg/100 ml, and 120 mg/100 ml of the Glycyrrhiza aqueous extract and 10 μg/ml, 20 μg/ml, and 40 μg/ml ribavirin in deionized water were evaluated for their toxicity and anti-viral activity in the embryonated eggs. Results: 60 mg/100 ml concentration of Glycyrrhiza extract did not produce any toxicity in the embryonated eggs and showed anti-viral activity against the virus. Similarly, 20 μg/ml ribavirin was non-toxic in the embryonated eggs and contained anti-viral activity. Conclusion: It may conclude from the presented study that 60 mg/100 ml Glycyrrhiza extract inhibits replication of Newcastle disease virus and is non-toxic in the embryonated eggs. So, Glycyrrhiza glabra extract may be further evaluated in future to determine the potentially active compounds for their anti-viral activity against Newcastle disease virus. Furthermore, the mechanism of action of these active phytochemicals as an antiviral agent would be helpful to elucidate the pathogenesis of the disease. PMID:24497736

  17. Gas-liquid chromatographic determination of kepone in field-collected avian tissues and eggs

    USGS Publications Warehouse

    Stafford, C.J.; Reichel, W.L.; Swineford, D.M.; Prouty, R.M.; Gay, M.L.

    1978-01-01

    A procedure is described for determining Kepone (decachlorooctahydro-1,3,4-metheno-2H-cyclobuta [cd] pentalene-2-one) residues in avian egg, liver, and tissue. Samples were extracted with benzene-isopropanol, and the extract was cleaned up with fuming H2SO4-concentrated H2SO4. Kepone was separated from organochlorine pesticides and polychlorinated biphenyls on a Florisil column and analyzed by electron capture gas-liquid chromatography (GLC). The average recovery from spiked tissues was 86%. The analyses performed on 14 bald eagle carcasses and livers, 3 bald eagle eggs, and 14 osprey eggs show measurable levels which indicate that Kepone accumulates in the tissues of fish-eating birds. Residues were confirmed by GLC-mass spectrometry.

  18. Evaluation of anthelmintic activity of Iris hookeriana against gastrointestinal nematodes of sheep.

    PubMed

    Tariq, K A; Chishti, M Z; Ahmad, F; Shawl, A S; Tantray, M A

    2008-06-01

    The objective of this study was to evaluate the anthelmintic efficacy of Iris hookeriana Linn. rhizome against gastrointestinal nematodes of sheep. A worm motility inhibition assay was used for in vitro study and a faecal egg count reduction assay was used for an in vivo study. The in vitro study revealed anthelmintic effects of crude aqueous extracts and crude ethanolic extracts on live Trichuris ovis worms (P < or = 0.05) as evident from their paralysis and/or death at 8 h after exposure. The aqueous extracts of I. hookeriana resulted in a mean worm motility inhibition of 54.0%, while ethanolic extracts resulted in a mean worm motility inhibition of 84.6%. The mean mortality index of aqueous extracts was 0.55, while for ethanolic extracts it was 0.85. The lethal concentration 50 for aqueous extracts was 0.45 mg ml- 1 and for ethanolic extracts it was 0.15 mg ml- 1. The in vivo anthelmintic activity of aqueous and ethanolic extracts of I. hookeriana in sheep naturally infected with mixed species of gastrointestinal nematodes demonstrated a maximum (45.62%) egg count reduction in sheep treated with ethanolic extracts at 2 g kg- 1 body weight on day 10 after treatment, closely followed by ethanolic extracts at 1 g kg- 1 body weight on day 10 after treatment (43.54% egg count reduction). The aqueous extracts resulted in a maximum of 31.53% reduction in faecal egg counts on day 10 after treatment with 1 g kg- 1 body weight. Thus ethanolic extracts exhibited greater anthelmintic activity under both in vitro and in vivo conditions; this could be due to the presence of alcohol-soluble active ingredients in I. hookeriana. From the present study it can be suggested that I. hookeriana rhizome exhibited significant anthelmintic activity against gastrointestinal nematodes of sheep and has the potential to contribute to the control of gastrointestinal nematode parasites of small ruminants.

  19. Oviposition in Delia platura (Diptera, Anthomyiidae): the role of volatile and contact cues of bean.

    PubMed

    Gouinguené, Sandrine P; Städler, Erich

    2006-07-01

    The choice of a suitable oviposition site by female insects is essential for survival of their progeny. Both olfactory and contact cues of the oviposition site may mediate this choice. The polyphagous Delia platura (Diptera: Anthomyiidae), a severe agricultural pest of numerous crops, lays eggs in the soil close to germinating seeds. Maggots feed upon the cotyledons. Only little is known about the cues guiding oviposition behavior. In this study, the effects of both olfactory and contact cues of beans (Phaseolus vulgaris) on oviposition of D. platura females were tested. Egg deposition on germinated beans was preferred to egg deposition on ungerminated beans or on beans in different postgerminating developmental stages. Olfactory cues of germinating beans alone stimulated female flies to lay eggs. Additional contact cues of germinating beans seemed to enhance the response, but the difference was not significant. Surface extracts of germinating beans sprayed on surrogate beans showed that both polar and nonpolar substances stimulated oviposition of D. platura flies. Gas chromatography-electroantennographic detection recordings of head space samples of germinating beans showed positive response of females to different compounds. We conclude that olfaction plays a major role when D. platura females are searching for oviposition sites. Volatile compounds released from germinating beans such as 4-hydroxy-4-methyl-2-pentanone, 1-hepten-3-one, 1-octen-3-ol, and 3-octanone should be considered as key compounds that mediate oviposition behavior. The use of different sensory modalities by closely related species of Delia is discussed.

  20. A comparative evaluation of six principal IgY antibody extraction methods.

    PubMed

    Ren, Hao; Yang, Wenjing; Thirumalai, Diraviyam; Zhang, Xiaoying; Schade, Rüdiger

    2016-03-01

    Egg yolk has been considered a promising source of antibodies. Our study was designed to compare six principal IgY extraction methods (water dilution, polyethylene glycol [PEG] precipitation, caprylic acid extraction, chloroform extraction, phenol extraction, and carrageenan extraction), and to assess their relative extraction efficiencies and the purity of the resulting antibodies. The results showed that the organic solvents (chloroform or phenol) minimised the lipid ratio in the egg yolk. The water dilution, PEG precipitation and caprylic acid extraction methods resulted in high yields, and antibodies purified with PEG and carrageenan exhibited high purity. Our results indicate that phenol extraction would be more suitable for preparing high concentrations of IgY for non-therapeutic usage, while the water dilution and carrageenan extraction methods would be more appropriate for use in the preparation of IgY for oral administration. 2016 FRAME.

  1. The c-mos proto-oncogene protein kinase turns on and maintains the activity of MAP kinase, but not MPF, in cell-free extracts of Xenopus oocytes and eggs.

    PubMed Central

    Nebreda, A R; Hunt, T

    1993-01-01

    During studies of the activation and inactivation of the cyclin B-p34cdc2 protein kinase (MPF) in cell-free extracts of Xenopus oocytes and eggs, we found that a bacterially expressed fusion protein between the Escherichia coli maltose-binding protein and the Xenopus c-mos protein kinase (malE-mos) activated a 42 kDa MAP kinase. The activation of MAP kinase on addition of malE-mos was consistent, whereas the activation of MPF was variable and failed to occur in some oocyte extracts in which cyclin A or okadaic acid activated both MPF and MAP kinase. In cases when MPF activation was transient, MAP kinase activity declined after MPF activity was lost, and MAP kinase, but not MPF, could be maintained at a high level by the presence of malE-mos. When intact oocytes were treated with progesterone, however, the activation of MPF and MAP kinase occurred simultaneously, in contrast to the behaviour of extracts. These observations suggest that one role of c-mos may be to maintain high MAP kinase activity in meiosis. They also imply that the activation of MPF and MAP kinase in vivo are synchronous events that normally rely on an agent that has still to be identified. Images PMID:8387916

  2. The role of HIRA and maternal histones in sperm nucleus decondensation in the gibel carp and color crucian carp.

    PubMed

    Zhao, Zhan-Ke; Li, Wei; Wang, Meng-Yu; Zhou, Li; Wang, Jia-Lin; Wang, Yu-Feng

    2011-02-01

    The histone H3.3 chaperone HIRA is essential for chromatin assembly during male pronucleus formation in Drosophila. However, the role of HIRA during fertilization in vertebrates remains unclear. The gibel carp (Carassius auratus gibelio) is a unique gynogenetic crucian carp (gyno-carp). Heterologous sperm nuclei cannot decondense when incorporated in the egg, thus the eggs produce a clonal lineage of all females by typical gynogenesis. In contrast, after entering the egg, homologous sperm can undergo decondensation and sexual reproduction is activated, which may produce both female and male offspring. Therefore, this fish is a useful model for studying the mechanisms of fertilization. Herein, we first compared HIRA expression during embryogenesis between gyno-carp and the gonochoristic color crucian carp (Carassius auratus; gono-carp). In gono-carp, a dramatic reduction of HIRA protein occurs shortly after fertilization, whereas HIRA protein is consistently expressed during embryogenesis of gyno-carp. Next, we used immunodepletion and an in vitro sperm decondensation system, and found that complete removal of HIRA inhibited sperm decondensation in both of the fish. Immunofluorescence localization showed that in the condensed sperm nuclei of gono-carp incubated in gyno-carp egg extracts, HIRA was detected, but neither the histone H2A variant H2af1o nor acetylated histone H4 was observed. These results suggest that HIRA may be a critical factor required for sperm nucleus decondensation, while the defect in deposition of some maternal histones in the sperm nucleus could be one reason why heterologous sperm cannot decondense in the gibel carp egg. Copyright © 2011 Wiley-Liss, Inc.

  3. Extraction and Analysis of V-Type Agents (VX, RVX, CVX, and VM) from Various Food Matrices by Ultra-Performance Liquid Chromatography-Time-Of-Flight Mass Spectrometry

    DTIC Science & Technology

    2015-12-01

    2% reduced fat milk, Egg Beaters egg whites, tomato sauce, and several meats, including hamburger meat (80% lean and 20% fat), hot dogs, chicken... Egg Whites and Tomato Sauce .....................................................3 2.3.3 Group 3: Hot Dogs, Chicken Nuggets, Turkey Deli Meat...Apple juice, orange juice, whole milk, 2% reduced fat milk, Egg Beaters processed egg whites, tomato sauce, precooked turkey deli meat (99% fat free

  4. In vitro ovicidal and larvicidal activity of methanolic leaf extract of Manihot esculenta (cassava) on susceptible and resistant strains of Trichostrongylus colubriformis.

    PubMed

    Al-Rofaai, A; Rahman, W A; Sulaiman, S F; Yahaya, Z S

    2012-11-23

    This study aimed to represent the first report of the ovicidal and larvicidal activity of the methanolic leaf extract of Manihot esculenta (cassava) against eggs and larvae of susceptible and resistant strains of Trichostrongylus colubriformis. As well as, to determine the total tannin compounds, antioxidant activity and toxicity of the extract. The egg hatch test was used to evaluate ovicidal activity against unembryonated eggs, whereas larval feeding inhibition assay and MTT-formazan assay were used to evaluate larvicidal activity against first (L(1)) and infective (L(3)) larvae, respectively. The results showed no significant differences were detected between the sensitivities of susceptible and resistant strains of T. colubriformis to the extract. Eggs, L(1) and L(3) were significantly affected (P<0.001) compared with negative control, and L(1) were more sensitive than the eggs and L(3). The total tannin compounds were investigated using tannin quantification assay and determined by 254.44 TAE/mg. The antioxidant activity was evaluated using the DPPH radical scavenging assay and the median inhibition concentration (IC(50)) was determined by 2.638 mg/ml. Acute oral toxicity at dose of 5,000 mg/kg, and sub-chronic oral toxicity at 500 and 1,000 mg/kg of the extract were observed in male and female Sprague-Dawley (SD) rats. The acute oral toxicity revealed that the median lethal dose (LD(50)) of methanolic extract of cassava leaves on SD rats was greater than 5,000 mg/kg, whereas the sub-chronic oral toxicity did not show observed adverse effects at 500 and 1,000 mg/kg per day for 28 days. In conclusion, the methanolic extract of cassava leaves has direct ovicidal and larvicidal activity against T. colubriformis strains with a safety margin for animals, and it may be potentially utilized as a source of natural antioxidants. Copyright © 2012 Elsevier B.V. All rights reserved.

  5. Tumor Suppression by BRCA-1: A Critical Role at DNA Replication Forks

    DTIC Science & Technology

    2006-10-01

    replication defect. We wished to test the hypothesis that BRCA1/BARD1 function during DNA replication supporting DNA transactions at replication forks. We...are using cell-free extracts derived from Xenopus laevis eggs that support: 1. Semi-conservative, cell-cycle regulated DNA replication ; 2. Many facets...complex assembles to chromatin in a DNA replication -dependent manner. Finally, we show that BRCA1/BARD1 loading to chromatin does not dramatically

  6. Assessment of in vitro and in vivo anthelminthic potential of extracts of Allium sativum bulb against naturally occurring ovine gastrointestinal nematodiosis.

    PubMed

    Kanojiya, Dharmendra; Shanker, Daya; Sudan, Vikrant; Jaiswal, Amit Kumar; Parashar, Rahul

    2015-01-01

    The rapid development of anthelminthic resistance has limited the success of traditional control programmes, thereby forcing researchers to search for ethno-veterinary alternatives. The objective is to assess the anthelminthic potential of various extracts of the bulb of Allium sativum in naturally infected sheep. In vitro anthelminthic activities of crude aqueous and methanolic extracts of the bulb of A. sativum were investigated against the egg (500 eggs/ml) and larvae of naturally infected sheep. The aqueous extract of A. sativum was also investigated for in vivo anthelminthic activity in three groups (n = 15 each) of naturally infected Chokla sheep with a negative control group receiving no treatment, a positive control group was given a single oral dose of albendazole at 7.5 mg/kg bodyweight, and a group administered a single oral dose of an aqueous extract at 5 g/animal. Data were analysed using the general linear model. Aqueous extract showed better efficacy in egg hatch assay and larval development test. However, in larval paralysis test, reverse trend was seen as methanolic extract was more potent than the aqueous counterpart. A significant amount of 57% faecal egg count reduction was observed in in vivo trail using the aqueous extract on day 21 post-treatment, although in initial stages it showed 30% and 83% effectiveness on days 7 and 14 post-treatment, respectively. No deleterious ill effect was found in any of the haematological and biochemical parameters. Bulb of A. sativum possesses good anthelminthic efficacy and further research is thereby warranted before recommending it for nematode control programme in ovines.

  7. Analysis of volatiles induced by oviposition of elm leaf beetle Xanthogaleruca luteola on Ulmus minor.

    PubMed

    Wegener, R; Schulz, S; Meiners, T; Hadwich, K; Hilker, M

    2001-03-01

    Egg deposition of the elm leaf beetle Xanthogaleruca luteola causes the emission of volatiles from its food plant, Ulmus minor. These volatiles are exploited by the egg parasitoid, Oomyzus gallerucae, to locate its host. In contrast to other tritrophic systems, the release of volatiles is not induced by feeding but by egg deposition. Previous investigations showed that the release is systemic and can be triggered by jasmonic acid. Comparison of headspace analysis revealed similarities in the blend of volatiles emitted following egg deposition and feeding. The mixture consists of more than 40 compounds; most of the substances are terpenoids. Leaves next to those carrying eggs emit fewer compounds. When treated with jasmonic acid, leaves emit a blend that consists almost exclusively of terpenoids. Dichloromethane extracts of leaves treated with jasmonic acid were also investigated. After separation of extracts of jasmonate induced elm leaves on silica, we obtained a fraction of terpenoid hydrocarbons that was attractive to the parasitoids. This indicates that jasmonic acid stimulates the production of terpenoid hydrocarbons that convey information of egg deposition to the parasitoid.

  8. Effect of the fungus Pochonia chlamydosporia on Echinostoma paraensei (Trematoda: Echinostomatidae).

    PubMed

    Lelis, Rosane Teixeira; Braga, Fabio Ribeiro; de Carvalho, Lorendane Millena; de Paula, Alessandra Teixeira; Araujo, Juliana Milani; Fausto, Mariana Costa; Junior, Arnaldo Maldonado; Rodrigues, João Victor Facchini; de Freitas Soares, Filippe Elias; Garcia, Juberlan Silva; de Araújo, Jackson Victor

    2014-11-01

    Echinostoma paraensei is a trematode of the genus Echinostoma that causes echinostomiasis in humans. The objectives of this study were to: evaluate the ovicidal activity of the nematophagous fungus Pochonia chlamydosporia (VC1 and VC4) on a solid medium 2% water-agar (2% WA) against E. paraensei eggs (assay A); evaluate ovicidal effect (destruction of eggs) of the isolate VC4 in supplemented culture media (assay B); and evaluate the ovicidal ability of the crude extract (VC4) on E. paraensei eggs (assay C). Eggs of E. paraensei (assay A) were placed in Petri dishes containing 2% WA with an isolate of the fungus P. chlamydosporia (VC1 and VC4) grown for 10 days, and without fungus as a control and evaluated regarding their destruction. In assay B, eggs of E. paraensei were placed in Petri dishes with different supplemented culture media and with VC4 isolate and the destruction of eggs was examined at the end of 25 days of interaction. In assay C, effects of the crude extract of P. chlamydosporia (VC4) on eggs were evaluated at the end of 7 days. In assay A, there was no difference (p>0.05) in ovicidal activity among the tested isolates (VC1 and VC4); however, the highest percentage for ovicidal activity (type 3 effect) was demonstrated by the isolate VC4. In assay B, the culture medium starch-agar showed the best results for the destruction of the eggs, with a percentage of 46.6% at the end of the assay. In assay C, the crude extract of VC4 was effective in the destruction of E. paraensei eggs, with a percentage reduction of 53%. The results of this study demonstrate that a rich culture medium with a greater availability of carbon and nitrogen may interfere directly in the predatory characteristics of ovicidal fungi. Copyright © 2014 Elsevier B.V. All rights reserved.

  9. Phosphorylation by Cdk1 Increases the Binding of Eg5 to Microtubules In Vitro and in Xenopus Egg Extract Spindles

    PubMed Central

    Cahu, Julie; Olichon, Aurelien; Hentrich, Christian; Schek, Henry; Drinjakovic, Jovana; Zhang, Cunjie; Doherty-Kirby, Amanda; Lajoie, Gilles; Surrey, Thomas

    2008-01-01

    Background Motor proteins from the kinesin-5 subfamily play an essential role in spindle assembly during cell division of most organisms. These motors crosslink and slide microtubules in the spindle. Kinesin-5 motors are phosphorylated at a conserved site by Cyclin-dependent kinase 1 (Cdk1) during mitosis. Xenopus laevis kinesin-5 has also been reported to be phosphorylated by Aurora A in vitro. Methodology/Principal Findings We investigate here the effect of these phosphorylations on kinesin-5 from Xenopus laevis, called Eg5. We find that phosphorylation at threonine 937 in the C-terminal tail of Eg5 by Cdk1 does not affect the velocity of Eg5, but strongly increases its binding to microtubules assembled in buffer. Likewise, this phosphorylation promotes binding of Eg5 to microtubules in Xenopus egg extract spindles. This enhancement of binding elevates the amount of Eg5 in spindles above a critical level required for bipolar spindle formation. We find furthermore that phosphorylation of Xenopus laevis Eg5 by Aurora A at serine 543 in the stalk is not required for spindle formation. Conclusions/Significance These results show that phosphorylation of Eg5 by Cdk1 has a direct effect on the interaction of this motor with microtubules. In egg extract, phosphorylation of Eg5 by Cdk1 ensures that the amount of Eg5 in the spindle is above a level that is required for spindle formation. This enhanced targeting to the spindle appears therefore to be, at least in part, a direct consequence of the enhanced binding of Eg5 to microtubules upon phosphorylation by Cdk1. These findings advance our understanding of the regulation of this essential mitotic motor protein. PMID:19079595

  10. Nuclear glycogen and glycogen synthase kinase 3.

    PubMed

    Ragano-Caracciolo, M; Berlin, W K; Miller, M W; Hanover, J A

    1998-08-19

    Glycogen is the principal storage form of glucose in animal cells. It accumulates in electron-dense cytoplasmic granules and is synthesized by glycogen synthase (GS), the rate-limiting enzyme of glycogen deposition. Glycogen synthase kinase-3 (GSK-3) is a protein kinase that phosphorylates GS. Two nearly identical forms of GSK-3 exist: GSK-3 alpha and GSK-3 beta. Both are constitutively active in resting cells and their activity can be modulated by hormones and growth factors. GSK-3 is implicated in the regulation of many physiological responses in mammalian cells by phosphorylating substrates including neuronal cell adhesion molecule, neurofilaments, synapsin I, and tau. Recent observations point to functions for glycogen and glycogen metabolism in the nucleus. GSK-3 phosphorylates several transcription factors, and we have recently shown that it modifies the major nuclear pore protein p62. It also regulates PK1, a protein kinase required for maintaining the interphase state and for DNA replication in cycling Xenopus egg extracts. Recently, glycogen was shown to be required for nuclear reformation in vitro using ovulated Xenopus laevis egg lysates. Because neither glycogen nor GSK-3 has been localized to the nuclear envelope or intranuclear sites, glycogen and GSK-3 activites were measured in rat liver nuclei and nuclear reformation extracts. Significant quantities of glycogen-like material co-purified with the rat-liver nuclear envelope. GSK-3 is also highly enriched in the glycogen pellet of egg extracts of Xenopus that is required for nuclear assembly in vitro. Based on the finding that enzymes of glycogen metabolism copurify with glycogen, we propose that glycogen may serve a structural role as a scaffold for nuclear assembly and sequestration of critical kinases and phosphatases in the nucleus. Copyright 1998 Academic Press.

  11. [Denaturation of egg antigens by cooking].

    PubMed

    Watanabe, Hiroko; Akaboshi, Chie; Sekido, Haruko; Tanaka, Kouki; Tanaka, Kazuko; Shimojo, Naoki

    2012-01-01

    Changes in egg protein contents by cooking were measured with an ELISA kit using Tris-HCl buffer in model foods including cake, meatballs, pasta and pudding made with whole egg, egg-white and egg-yolk. The egg protein contents were lowest in the deep-fried model foods of cakes and meatballs. Ovalbumin (OVA) was undetectable (<1 µg/g) and ovomucoid (OVM) was lowest in pouched meatballs, suggesting that processing temperature and uniform heat-treatment affect the detection of egg protein. Furthermore, egg protein contents were below 6 µg/g in the pouched meatballs and pasta made with egg-yolk, and OVA and OVM were not detected by Western blotting analysis with human IgE from patients' serum. On the other hand, processed egg proteins were detected with an ELISA kit using a surfactant and reductant in the extract buffer.

  12. Phosphatidylcholine from "Healthful" Egg Yolk Varieties: An Organic Laboratory Experience

    NASA Astrophysics Data System (ADS)

    Hodges, Linda C.

    1995-12-01

    I have added an investigative element to a popular undergraduate experiment. the characterization of phosphatidylcholine (PC) from egg yolks. Varieties of eggs are commercially available which have been obtained from chickens fed a diet containing no animal fat. Presumably, less saturated fat in the diet of the chickens could be reflected in the fatty acid composition of various classes of biological lipids, including phospholipids, in the eggs from these chickens. PC is extracted using conventional methods, the extract is further purified by chromatography on silicic acid, and the column fractions are assayed for the presence and purity of PC by TLC. Fractions containing pure PC are pooled, concentrated, hydrolyzed, and esterified to obtain the fatty acid methyl esters (FAME) which are identified by GLC. Comparing FAMEs derived from PC of yolks of regular eggs to those obtained from the other special brands adds a novel twist to the students' work and generates greater student interest and involvement in both the interpretation of data than a simple isolation of a biological compound alone evokes.

  13. Determination of alkylphenol and alkylphenolethoxylates in biota by liquid chromatography with detection by tandem mass spectrometry and fluorescence spectroscopy

    USGS Publications Warehouse

    Schmitz-Afonso, I.; Loyo-Rosales, J.E.; de la Paz Aviles, M.; Rattner, B.A.; Rice, C.P.

    2003-01-01

    A quantitative method for the simultaneous determination of octylphenol, nonylphenol and the corresponding ethoxylates (1 to 5) in biota is presented. Extraction methods were developed for egg and fish matrices based on accelerated solvent extraction followed by a solid-phase extraction cleanup, using octadecylsilica or aminopropyl cartridges. Identification and quantitation were accomplished by liquid chromatography-electrospray tandem mass spectrometry (LC-MS-MS) and compared to the traditional liquid chromatography with fluorescence spectroscopy detection. LC-MS-MS provides high sensitivity and specificity required for these complex matrices and an accurate quantitation with the use of 13C-labeled internal standards. Quantitation limits by LC-MS-MS ranged from 4 to 12 ng/g in eggs, and from 6 to 22 ng/g in fish samples. These methods were successfully applied to osprey eggs from the Chesapeake Bay and fish from the Great Lakes area. Total levels found in osprey egg samples were up to 18 ng/g wet mass and as high as 8.2 ug/g wet mass in the fish samples.

  14. Arabidopsis MYC Transcription Factors Are the Target of Hormonal Salicylic Acid/Jasmonic Acid Cross Talk in Response to Pieris brassicae Egg Extract1[OPEN

    PubMed Central

    Schmiesing, André; Gouhier-Darimont, Caroline

    2016-01-01

    Arabidopsis (Arabidopsis thaliana) plants recognize insect eggs and activate the salicylic acid (SA) pathway. As a consequence, expression of defense genes regulated by the jasmonic acid (JA) pathway is suppressed and larval performance is enhanced. Cross talk between defense signaling pathways is common in plant-pathogen interactions, but the molecular mechanism mediating this phenomenon is poorly understood. Here, we demonstrate that egg-induced SA/JA antagonism works independently of the APETALA2/ETHYLENE RESPONSE FACTOR (AP2/ERF) transcription factor ORA59, which controls the ERF branch of the JA pathway. In addition, treatment with egg extract did not enhance expression or stability of JASMONATE ZIM-domain transcriptional repressors, and SA/JA cross talk did not involve JASMONATE ASSOCIATED MYC2-LIKEs, which are negative regulators of the JA pathway. Investigating the stability of MYC2, MYC3, and MYC4, three basic helix-loop-helix transcription factors that additively control jasmonate-related defense responses, we found that egg extract treatment strongly diminished MYC protein levels in an SA-dependent manner. Furthermore, we identified WRKY75 as a novel and essential factor controlling SA/JA cross talk. These data indicate that insect eggs target the MYC branch of the JA pathway and uncover an unexpected modulation of SA/JA antagonism depending on the biological context in which the SA pathway is activated. PMID:26884488

  15. Arabidopsis MYC Transcription Factors Are the Target of Hormonal Salicylic Acid/Jasmonic Acid Cross Talk in Response to Pieris brassicae Egg Extract.

    PubMed

    Schmiesing, André; Emonet, Aurélia; Gouhier-Darimont, Caroline; Reymond, Philippe

    2016-04-01

    Arabidopsis (Arabidopsis thaliana) plants recognize insect eggs and activate the salicylic acid (SA) pathway. As a consequence, expression of defense genes regulated by the jasmonic acid (JA) pathway is suppressed and larval performance is enhanced. Cross talk between defense signaling pathways is common in plant-pathogen interactions, but the molecular mechanism mediating this phenomenon is poorly understood. Here, we demonstrate that egg-induced SA/JA antagonism works independently of the APETALA2/ETHYLENE RESPONSE FACTOR (AP2/ERF) transcription factor ORA59, which controls the ERF branch of the JA pathway. In addition, treatment with egg extract did not enhance expression or stability of JASMONATE ZIM-domain transcriptional repressors, and SA/JA cross talk did not involve JASMONATE ASSOCIATED MYC2-LIKEs, which are negative regulators of the JA pathway. Investigating the stability of MYC2, MYC3, and MYC4, three basic helix-loop-helix transcription factors that additively control jasmonate-related defense responses, we found that egg extract treatment strongly diminished MYC protein levels in an SA-dependent manner. Furthermore, we identified WRKY75 as a novel and essential factor controlling SA/JA cross talk. These data indicate that insect eggs target the MYC branch of the JA pathway and uncover an unexpected modulation of SA/JA antagonism depending on the biological context in which the SA pathway is activated. © 2016 American Society of Plant Biologists. All Rights Reserved.

  16. Oviposition and Embryotoxicity of Indigofera suffruticosa on Early Development of Aedes aegypti (Diptera: Culicidae)

    PubMed Central

    Vieira, Jeymesson Raphael Cardoso; Leite, Roberta Maria Pereira; Lima, Izabela Rangel; Navarro, Daniela do Amaral Ferraz; Bianco, Everson Miguel; Leite, Sônia Pereira

    2012-01-01

    Aqueous extract of Indigofera suffruticosa leaves obtained by infusion was used to evaluate the oviposition, its effect on development of eggs and larvae, and morphological changes in larvae of Aedes aegypti. The bioassays were carried out with aqueous extract in different concentrations on eggs, larvae, and female mosquitoes, and the morphological changes were observed in midgut of larvae. The extract showed repellent activity on A. aegypti mosquitoes, reducing significantly the egg laying by females with control substrate (343 (185–406)) compared with the treated substrate (88 (13–210)). No eclosion of A. aegypti eggs at different concentrations studied was observed. The controleclodedin 35%. At concentration of 250 μg/mL, 93.3% of larvae remained in the second instar of development and at concentrations of 500, 750, and 1000 μg/mL the inhibitory effect was lower with percentages of 20%, 53.3%, and 46.6%, respectively. Morphological changes like disruption on the peritrophic envelope (PE), discontinued underlying epithelium, increased gut lumen, and segments with hypertrophic aspects were observed in anterior region of medium midgut of larvae of A. aegypti. The results showed repellent activity, specific embryotoxicity, and general growth retardation in A. aegypti by medium containing aqueous extract of I. suffruticosa leaves. PMID:21822443

  17. Embryotoxicity of Great Lakes lake trout extracts to developing rainbow trout

    USGS Publications Warehouse

    Wright, Peggy J.; Tillitt, Donald E.

    1999-01-01

    Planar halogenated hydrocarbons (PHHs), such as polychlorinated dibenzo-p-dioxins, dibenzofurans, and biphenyls are present in aquatic systems, and are known to produce adverse effects in fish. This study investigated the embryotoxicity of PHH mixtures through the nanoinjection of environmental extracts into newly fertilized eggs from two strains of rainbow trout. Organic extracts were obtained from whole adult lake trout collected from Lake Michigan in 1988 and Lake Superior in 1994. The graded doses of the final extracts used for injection were quantified as 2,3,7,8-tetrachlorodibenzo-p-dioxin toxic-equivalents (TEQs) based on the concentrations of dioxins, furans and non-o-PCBs in each, and as equivalent amounts found in the eggs of the original lake trout (eggEQ). Total TEQs in the lake trout were 14.7 pg TEQ/g in the Lake Michigan sample and 7.3 pg TEQ/g in the Lake Superior sample. The extract of the Lake Michigan lake trout was embryotoxic to rainbow trout; LD50 values were 35 eggEQ (15–90, 95% F.L.) in the Arlee strain and 14 eggEQ (5–99, 95% F.L.) in the Erwin strain of rainbow trout. The LD50 values of the Lake Michigan extract in either of these strains of rainbow trout fall within the actual range of TCDD LD50values based on TEQs. This indicates that an additive model of toxicity is appropriate to quantify PHHs in relation to early life stage mortality in fish. Gross lesions characteristic of exposure to PHHs (i.e. yolk-sac edema, craniofacial deformities, and hemorrhaging) increased in a dose-related manner. The lowest observable adverse effect concentrations (LOAEC) for these gross lesions and cumulative mortalities suggests that current concentrations of PHHs in lake trout from Lake Michigan are above a threshold for adverse effects and these compounds may have implications on the lack of recruitment in certain Great Lakes lake trout populations.

  18. Is there a negative association between the content of condensed tannins, total phenols, and total tannins of tropical plant extracts and in vitro anthelmintic activity against Haemonchus contortus eggs?

    PubMed

    Castañeda-Ramírez, G S; Torres-Acosta, J F J; Sandoval-Castro, C A; González-Pech, P G; Parra-Tabla, V P; Mathieu, C

    2017-12-01

    In vitro studies using plant extracts suggest a relationship between their polyphenol contents and their anthelmintic (AH) activity against Haemonchus contortus. High polyphenol content appears to increase the efficacy of plant extracts against H. contortus as assessed by the larval exsheathment inhibition assay (LEIA) while appearing to reduce the AH efficacy measured using the egg hatch assay (EHA). In addition, some plants lack AH activity. Therefore, the present study investigated the relationship between the contents of condensed tannins (CT), total phenols (TP), and total tannins (TT) in methanol:water extracts (70:30) obtained from ten tropical plant species consumed by small ruminants as well as their AH activity against H. contortus evaluated by LEIA and EHA. Extracts of Acacia collinsii, Lysiloma latisiliquum, Havardia albicans, Senegalia gaumeri, Mimosa bahamensis, Piscidia piscipula, Acacia pennatula, Gymnopodium floribundum, Leucaena leucocephala, and Bunchosia swartziana were examined. Positive correlations were found between the effective concentration 50% (EC 50 ) (EHA) of extracts and their CT (r = 0.6809, P < 0.05, n = 10) and TP (r = 0.9152, P < 0.05, n = 10) content, suggesting that their concentration negatively affected AH activity against eggs. Based on the LEIA, there was no significant association between the EC 50 and the CT, TP, or TT of all extracts evaluated. Thus, if sheep and goats consume a complex feed mixture with high amounts of CT, TP, and TT, it might be difficult to observe an AH effect against H. contortus egg hatching. However, the AH effect upon L 3 establishment might be feasible.

  19. Effect of lipid extraction on analyses of stable carbon and stable nitrogen isotopes in coastal organisms of the Aleutian archipelago

    USGS Publications Warehouse

    Ricca, M.A.; Miles, A.K.; Anthony, R.G.; Deng, X.; Hung, S.S.O.

    2007-01-01

    We tested whether extracting lipids reduced confounding variation in ??13C and ??15N values by analyzing paired lipid-extracted (LE) and non-lipid-extracted (NLE) samples of bald eagle (Haliaeetus leucocephalus (L., 1766)) whole eggs, muscle tissue from nine seabird and one terrestrial bird species, muscle tissue from four marine fish species, and blue mussels (Mytilus edulis L., 1758) collected from the Aleutian archipelago, Alaska. Lipid extraction significantly increased ??13C by an average of 2.0??? in whole eggs, 0.8??? in avian muscle, 0.2??? in fish muscle, and 0.6??? in blue mussels. Lower ??13C values in NLE samples covaried positively with lipid content across all sample types. Lower ??13C values in NLE samples were not correlated with lipid content within bald eagle eggs and blue mussels, but covaried positively with percent lipid in avian and fish muscles. Neither lipid extraction nor percent lipid significantly changed ??15N values for any sample type. Lower ??13C values in most NLE avian and fish muscle tissues should not confound interpretation of pelagic versus nearshore sources of primary production, but lipid extraction may be necessary when highly precise estimates of ??13C are needed. Lipid extraction may not be necessary when only ??15N is of interest. ?? 2007 NRC.

  20. Evaluation of Anthelmintic Activity and Composition of Pumpkin (Cucurbita pepo L.) Seed Extracts-In Vitro and in Vivo Studies.

    PubMed

    Grzybek, Maciej; Kukula-Koch, Wirginia; Strachecka, Aneta; Jaworska, Aleksandra; Phiri, Andrew M; Paleolog, Jerzy; Tomczuk, Krzysztof

    2016-09-01

    A significant number of studies report growing resistance in nematodes thriving in both humans and livestock. This study was conducted to evaluate the in vitro and in vivo anthelmintic efficiency of Curcubita pepo (C. pepo) L. hot water extract (HWE), cold water extract (CWE) or ethanol extract (ETE) on two model nematodes: Caenorhabditis elegans (C. elegans) and Heligmosoides bakeri (H. bakeri). Raman, IR and LC-MS spectroscopy analyses were performed on the studied plant material to deliver qualitative and quantitative data on the composition of the obtained extracts: ETE, HWE and CWE. The in vitro activity evaluation showed an impact of C. pepo extracts on C. elegans and different developmental stages of H. bakeri. The following in vivo experiments on mice infected with H. bakeri confirmed inhibitory properties of the most active pumpkin extract selected by the in vitro study. All of the extracts were found to contain cucurbitine, aminoacids, fatty acids, and-for the first time-berberine and palmatine were identified. All C. pepo seed extracts exhibited a nematidicidal potential in vitro, affecting the survival of L1 and L2 H. bakeri larvae. The ETE was the strongest and demonstrated a positive effect on H. bakeri eggs hatching and marked inhibitory properties against worm motility, compared to a PBS control. No significant effects of pumpkin seed extracts on C. elegans integrity or motility were found. The EtOH extract in the in vivo studies showed anthelmintic properties against both H. bakeri fecal egg counts and adult worm burdens. The highest egg counts reduction was observed for the 8 g/kg dose (IC50 against H. bakeri = 2.43; 95% Cl = 2.01-2.94). A decrease in faecal egg counts (FEC) was accompanied by a significant reduction in worm burden of the treated mice compared to the control group. Pumpkin seed extracts may be used to control of Gastrointestinal (G.I.) nematode infections. This relatively inexpensive alternative to the currently available chemotherapeutic should be considered as a novel drug candidate in the nearest future.

  1. Development of multiclass methods for drug residues in eggs: hydrophilic solid-phase extraction cleanup and liquid chromatography/tandem mass spectrometry analysis of tetracycline, fluoroquinolone, sulfonamide, and beta-lactam residues.

    PubMed

    Heller, David N; Nochetto, Cristina B; Rummel, Nathan G; Thomas, Michael H

    2006-07-26

    A method was developed for detection of a variety of polar drug residues in eggs via liquid chromatography/tandem mass spectrometry (LC/MS/MS) with electrospray ionization (ESI). A total of twenty-nine target analytes from four drug classes-sulfonamides, tetracyclines, fluoroquinolones, and beta-lactams-were extracted from eggs using a hydrophilic-lipophilic balance polymer solid-phase extraction (SPE) cartridge. The extraction technique was developed for use at a target concentration of 100 ng/mL (ppb), and it was applied to eggs containing incurred residues from dosed laying hens. The ESI source was tuned using a single, generic set of tuning parameters, and analytes were separated with a phenyl-bonded silica cartridge column using an LC gradient. In a related study, residues of beta-lactam drugs were not found by LC/MS/MS in eggs from hens dosed orally with beta-lactam drugs. LC/MS/MS performance was evaluated on two generations of ion trap mass spectrometers, and key operational parameters were identified for each instrument. The ion trap acquisition methods could be set up for screening (a single product ion) or confirmation (multiple product ions). The lower limit of detection for screening purposes was 10-50 ppb (sulfonamides), 10-20 ppb (fluoroquinolones), and 10-50 ppb (tetracyclines), depending on the drug, instrument, and acquisition method. Development of this method demonstrates the feasibility of generic SPE, LC, and MS conditions for multiclass LC/MS residue screening.

  2. Long-term effect on in vitro cloning efficiency after treatment of somatic cells with Xenopus egg extract in the pig.

    PubMed

    Liu, Ying; Ostrup, Olga; Li, Rong; Li, Juan; Vajta, Gábor; Kragh, Peter M; Schmidt, Mette; Purup, Stig; Hyttel, Poul; Klærke, Dan; Callesen, Henrik

    2014-08-01

    In somatic cell nuclear transfer (SCNT), donor cell reprogramming is considered as a biologically important and vulnerable event. Various donor cell pre-treatments with Xenopus egg extracts can promote reprogramming. Here we investigated if the reprogramming effect of one treatment with Xenopus egg extract on donor cells was maintained for several cell passages. The extract treatment resulted in increased cell-colony formation from early passages in treated porcine fibroblasts (ExTES), and increased development of cloned embryos. Partial dedifferentiation was observed in ExTES cells, shown as a tendency towards upregulation of NANOG, c-MYC and KLF-4 and downregulation of DESMIM compared with ExTES at Passage 2. Compared with our routine SCNT, continuously increased development of cloned embryos was observed in the ExTES group, and ExTES cloned blastocysts displayed hypermethylated DNA patterns and hypermethylation of H3K4me3 and H3K27me3 in ICM compared with TE. All seven recipients became pregnant after transferral of ExTES cloned embryos and gave birth to 7-22 piglets per litter (average 12). In conclusion, our results demonstrate that one treatment of porcine fibroblasts with Xenopus egg extract can result in long-term increased ability of the cells to promote their in vitro function in subsequent SCNT. Finally these cells can also result in successful development of cloned embryos to term.

  3. Detection of Torque teno midi virus/Small anellovirus (TTMDV/SAV) in the sera of domestic village chickens and its vertical transmission from hen to eggs

    PubMed Central

    Bouzari, M; Salmanizadeh, Sh

    2015-01-01

    Although the infection of different animals and non-human primates with other members of Anelloviridae have already been reported there is no report about infection of animals with Torque teno midi virus/Small anellovirs (TTMDV/SAV). The aim of this study was to detect the virus in domestic village chickens. Blood samples were collected from 79 domestic village chickens in Isfahan. Blood samples of five adult laying hens and one cockerel were collected in three consecutive weeks (days 1, 8 and 14) as experimental chickens. Ten eggs were randomly collected from the eggs laid during days 12 to 17 and thin and thick egg whites and yolk samples were collected aseptically. After DNA extraction Nested-PCR was performed using SMAs/SMAr primers. In PCR, 431 bp and 441 bp products were detected. The detected bands were extracted and sequenced. Totally 26 out of 79 (32.9%) of the blood samples were positive for the virus. The frequency of the infection of the different parts of the eggs tested was 76%. For the first time TTMDV/SAV was detected in domestic village chickens which also vertically transmitted to eggs. PMID:27175162

  4. Detection of Torque teno midi virus/Small anellovirus (TTMDV/SAV) in the sera of domestic village chickens and its vertical transmission from hen to eggs.

    PubMed

    Bouzari, M; Salmanizadeh, Sh

    2015-01-01

    Although the infection of different animals and non-human primates with other members of Anelloviridae have already been reported there is no report about infection of animals with Torque teno midi virus/Small anellovirs (TTMDV/SAV). The aim of this study was to detect the virus in domestic village chickens. Blood samples were collected from 79 domestic village chickens in Isfahan. Blood samples of five adult laying hens and one cockerel were collected in three consecutive weeks (days 1, 8 and 14) as experimental chickens. Ten eggs were randomly collected from the eggs laid during days 12 to 17 and thin and thick egg whites and yolk samples were collected aseptically. After DNA extraction Nested-PCR was performed using SMAs/SMAr primers. In PCR, 431 bp and 441 bp products were detected. The detected bands were extracted and sequenced. Totally 26 out of 79 (32.9%) of the blood samples were positive for the virus. The frequency of the infection of the different parts of the eggs tested was 76%. For the first time TTMDV/SAV was detected in domestic village chickens which also vertically transmitted to eggs.

  5. Iodine-xenon studies of Allende inclusions - Eggs and the Pink Angel

    NASA Technical Reports Server (NTRS)

    Swindle, T. D.; Caffee, M. W.; Hohenberg, C. M.

    1988-01-01

    The I-Xe systems of six Allende inclusions (five Eggs and the Pink Angel) appear to have been altered by nonnebular secondary processes. Evidence for this includes temperature-ordered variations in the initial I isotopic composition within several objects (with older apparent I-Xe ages associated with higher extraction temperatures) and the absence of primitive I-Xe ages. The span of apparent ages seen in Allende objects (10 Myr or more) is probably too long to reflect any nebular process, so at least some alteration probably occurred ont the parent body. The range in initial (Pu-244)/(U-238) ratios for the Eggs (0.003-0.014) includes the current best estimates of the bulk solar system value (0.004-0.007). For Egg 3, the Pu/U ratio varies by a factor of two between extractions, probably the result of fractionation of Pu from U among different phases.

  6. Evaluation of Anthelmintic Activity and Composition of Pumpkin (Cucurbita pepo L.) Seed Extracts—In Vitro and in Vivo Studies

    PubMed Central

    Grzybek, Maciej; Kukula-Koch, Wirginia; Strachecka, Aneta; Jaworska, Aleksandra; Phiri, Andrew M.; Paleolog, Jerzy; Tomczuk, Krzysztof

    2016-01-01

    A significant number of studies report growing resistance in nematodes thriving in both humans and livestock. This study was conducted to evaluate the in vitro and in vivo anthelmintic efficiency of Curcubita pepo (C. pepo) L. hot water extract (HWE), cold water extract (CWE) or ethanol extract (ETE) on two model nematodes: Caenorhabditis elegans (C. elegans) and Heligmosoides bakeri (H. bakeri). Methods: Raman, IR and LC-MS spectroscopy analyses were performed on the studied plant material to deliver qualitative and quantitative data on the composition of the obtained extracts: ETE, HWE and CWE. The in vitro activity evaluation showed an impact of C. pepo extracts on C. elegans and different developmental stages of H. bakeri. The following in vivo experiments on mice infected with H. bakeri confirmed inhibitory properties of the most active pumpkin extract selected by the in vitro study. All of the extracts were found to contain cucurbitine, aminoacids, fatty acids, and-for the first time-berberine and palmatine were identified. All C. pepo seed extracts exhibited a nematidicidal potential in vitro, affecting the survival of L1 and L2 H. bakeri larvae. The ETE was the strongest and demonstrated a positive effect on H. bakeri eggs hatching and marked inhibitory properties against worm motility, compared to a PBS control. No significant effects of pumpkin seed extracts on C. elegans integrity or motility were found. The EtOH extract in the in vivo studies showed anthelmintic properties against both H. bakeri fecal egg counts and adult worm burdens. The highest egg counts reduction was observed for the 8 g/kg dose (IC50 against H. bakeri = 2.43; 95% Cl = 2.01–2.94). A decrease in faecal egg counts (FEC) was accompanied by a significant reduction in worm burden of the treated mice compared to the control group. Conclusions: Pumpkin seed extracts may be used to control of Gastrointestinal (G.I.) nematode infections. This relatively inexpensive alternative to the currently available chemotherapeutic should be considered as a novel drug candidate in the nearest future. PMID:27598135

  7. Egg extract of apple snail for eco-friendly synthesis of silver nanoparticles and their antibacterial activity.

    PubMed

    Janthima, Ratima; Khamhaengpol, Arunrat; Siri, Sineenat

    2018-03-01

    Green synthesis of silver nanoparticles (AgNPs) provides the alternative method with cost effectiveness and the eco-friendly process by using natural biomolecules as reducing and stabilizing agents. Alternative to the most studies of plant extracts, this work demonstrated a use of egg extract of apple snail (Pomacea canaliculata) for an eco-friendly production of AgNPs. The extract contained at least six proteins with the molecular weight in a range of 24-65 kDa that exhibited the reducing activity. The dispersive AgNPs were produced in the reaction containing only the extract and silver nitrate, as determined by the characteristic surface plasmon resonance peak of silver at 412 nm. The synthesized AgNPs were spherical with the average diameter of 9.0 ± 5.9 nm. The X-ray diffraction pattern and selected area electron diffraction (SAED) analyses confirmed the face-cubic centre (fcc) unit cell structure of AgNPs. The synthesized AgNPs exhibited the antibacterial activity against both Gram-positive Staphylococcus aureus and Gram-negative Escherichia coli. Results of this work clearly showed the potential use of the egg extract of apple snail for a green synthesis of small size AgNPs exhibiting the antibacterial activity.

  8. EGG: Empirical Galaxy Generator

    NASA Astrophysics Data System (ADS)

    Schreiber, C.; Elbaz, D.; Pannella, M.; Merlin, E.; Castellano, M.; Fontana, A.; Bourne, N.; Boutsia, K.; Cullen, F.; Dunlop, J.; Ferguson, H. C.; Michałowski, M. J.; Okumura, K.; Santini, P.; Shu, X. W.; Wang, T.; White, C.

    2018-04-01

    The Empirical Galaxy Generator (EGG) generates fake galaxy catalogs and images with realistic positions, morphologies and fluxes from the far-ultraviolet to the far-infrared. The catalogs are generated by egg-gencat and stored in binary FITS tables (column oriented). Another program, egg-2skymaker, is used to convert the generated catalog into ASCII tables suitable for ingestion by SkyMaker (ascl:1010.066) to produce realistic high resolution images (e.g., Hubble-like), while egg-gennoise and egg-genmap can be used to generate the low resolution images (e.g., Herschel-like). These tools can be used to test source extraction codes, or to evaluate the reliability of any map-based science (stacking, dropout identification, etc.).

  9. Multi-residue determination of 115 veterinary drugs and pharmaceutical residues in milk powder, butter, fish tissue and eggs using liquid chromatography-tandem mass spectrometry.

    PubMed

    Dasenaki, Marilena E; Thomaidis, Nikolaos S

    2015-06-23

    A simple and sensitive multi-residue method for the determination of 115 veterinary drugs and pharmaceuticals, belonging in more than 20 different classes, in butter, milk powder, egg and fish tissue has been developed. The method involves a simple generic solid-liquid extraction step (solvent extraction, SE) with 0.1% formic acid in aqueous solution of EDTA 0.1% (w/v)-acetonitrile (ACN)-methanol (MeOH) (1:1:1, v/v) with additional ultrasonic-assisted extraction. Precipitation of lipids and proteins was promoted by subjecting the extracts at very low temperature (-23°C) for 12h. Further cleanup with hexane ensures fat removal from the matrix. Analysis was performed by liquid chromatography coupled with electrospray ionization and tandem mass spectrometry (LC-ESI-MS/MS). Two separate runs were performed for positive and negative ionization in multiple reaction monitoring mode (MRM). Particular attention was devoted to extraction optimization: different sample-to-extracting volume ratios, different concentrations of formic acid in the extraction solvent and different ultrasonic extraction temperatures were tested in butter, egg and milk powder samples. The method was also applied in fish tissue samples. It was validated, on the basis of international guidelines, for all four matrices. Quantitative analysis was performed by means of standard addition calibration. For over 80% of the analytes, the recoveries were between 50% and 120% in all matrices studied, with RSD values in the range of 1-18%. Limits of detection (LODs) and quantification (LOQs) ranged from 0.008 μg kg(-1) (oxfendazole in butter) to 3.15 μg kg(-1) (hydrochlorthiazide in egg). The evaluated method provides reliable screening, quantification, and identification of 115 veterinary drug and pharmaceutical residues in foods of animal origin and has been successfully applied in real samples. Copyright © 2015 Elsevier B.V. All rights reserved.

  10. HPLC Quantification of astaxanthin and canthaxanthin in Salmonidae eggs.

    PubMed

    Tzanova, Milena; Argirova, Mariana; Atanasov, Vasil

    2017-04-01

    Astaxanthin and canthaxanthin are naturally occurring antioxidants referred to as xanthophylls. They are used as food additives in fish farms to improve the organoleptic qualities of salmonid products and to prevent reproductive diseases. This study reports the development and single-laboratory validation of a rapid method for quantification of astaxanthin and canthaxanthin in eggs of rainbow trout (Oncorhynchus mykiss) and brook trout (Salvelinus fontinalis М.). An advantage of the proposed method is the perfect combination of selective extraction of the xanthophylls and analysis of the extract by high-performance liquid chromatography and photodiode array detection. The method validation was carried out in terms of linearity, accuracy, precision, recovery and limits of detection and quantification. The method was applied for simultaneous quantification of the two xanthophylls in eggs of rainbow trout and brook trout after their selective extraction. The results show that astaxanthin accumulations in salmonid fish eggs are larger than those of canthaxanthin. As the levels of these two xanthophylls affect fish fertility, this method can be used to improve the nutritional quality and to minimize the occurrence of the M74 syndrome in fish populations. Copyright © 2016 John Wiley & Sons, Ltd.

  11. Quantitative analysis of amoxicillin, its major metabolites and ampicillin in eggs by liquid chromatography combined with electrospray ionization tandem mass spectrometry.

    PubMed

    Sun, Lirui; Jia, Longfei; Xie, Xing; Xie, Kaizhou; Wang, Jianfeng; Liu, Jianyu; Cui, Lulu; Zhang, Genxi; Dai, Guojun; Wang, Jinyu

    2016-02-01

    In this present study, we developed a simple, rapid and specific method for the quantitative analysis of the contents of amoxicillin (AMO), AMO metabolites and ampicillin (AMP) in eggs. This method uses a simple liquid-liquid extraction with acetonitrile followed by liquid chromatography-tandem mass spectrometry (LC-MS/MS). The optimized method has been validated according to requirements defined by the European Union and Food and Drug Administration. Extraction recoveries of the target compounds from the egg at 5, 10 and 25 μg/kg were all higher than 80%, with relative standard deviations not exceeding 10.00%. The limits of quantification in eggs were below the maximum residue limits (MRLs). The decision limits (CCα) ranged between 11.1 and 11.5 μg/kg, while detection capabilities (CCβ) from 12.1 to 13.0 μg/kg. These values were very close to the corresponding MRLs. Finally, the new approach was successfully verified for the quantitative determination of these analytes in 40 commercial eggs from local supermarkets. Copyright © 2015 Elsevier Ltd. All rights reserved.

  12. Impact of casein and egg white proteins on the structure of wheat gluten-based protein-rich food.

    PubMed

    Wouters, Arno G B; Rombouts, Ine; Lagrain, Bert; Delcour, Jan A

    2016-02-01

    There is a growing interest in texturally and nutritionally satisfying vegetable alternatives to meat. Wheat gluten proteins have unique functional properties but a poor nutritional value in comparison to animal proteins. This study investigated the potential of egg white and bovine milk casein with well-balanced amino acid composition to increase the quality of wheat gluten-based protein-rich foods. Heating a wheat gluten (51.4 g)-water (100.0 mL) blend for 120 min at 100 °C increased its firmness less than heating a wheat gluten (33.0 g)-freeze-dried egg white (16.8 g)-water (100.0 mL) blend. In contrast, the addition of casein to the gluten-water blend negatively impacted firmness after heating. Firmness was correlated with loss of protein extractability in sodium dodecyl sulfate containing medium during heating, which was higher with egg white than with casein. Even more, heat-induced polymerization of the gluten-water blend with egg white but not with casein was greater than expected from the losses in extractability of gluten and egg white on their own. Structure formation was favored by mixing gluten with egg white but not with casein. These observations were linked to the intrinsic polymerization behavior of egg white and casein, but also to their interaction with gluten. Thus not all nutritionally suitable proteins can be used for enrichment of gluten-based protein-rich foods. © 2015 Society of Chemical Industry.

  13. In vivo evaluation of the antiviral activity of Cajanus cajan on measles virus.

    PubMed

    Nwodo, U U; Ngene, A A; Iroegbu, C U; Onyedikachi, O A L; Chigor, V N; Okoh, A I

    2011-09-01

    Cajanus cajan, a tropical shrub, serves as source of food and traditional medicines. The evaluation of aqueous and ethanol extracts for activity against measles virus and toxicity to embryonated chicken eggs was carried out in this study. In vivo and in vitro assay techniques using embryonated chicken eggs and tissue culture (Hep-2 cell lines) as media for both virus cultivation and anti-virus assay showed that a hot-water extract yielded higher activity against measles virus. The hot-water extract of the stem yielded a Log(2) titre of 0.1 for the in vivo assay and an inhibition of cytopathic effect (CPE) in Hep-2 cells by 100% for the in vitro assay. At all concentrations of the extracts, there was a lowering of virus concentration (p = 0.05), indicated by hemagglutination (HA) titration, which is the advantage of HA titration over the tissue culture technique using CPE. This study validates embryonated chicken eggs as suitable media for anti-virus assay and the use of C. cajan in the treatment of some diseases of viral origin.

  14. Lipid oxidation in baked products: impact of formula and process on the generation of volatile compounds.

    PubMed

    Maire, Murielle; Rega, Barbara; Cuvelier, Marie-Elisabeth; Soto, Paola; Giampaoli, Pierre

    2013-12-15

    This paper investigates the effect of ingredients on the reactions occurring during the making of sponge cake and leading to the generation of volatile compounds related to flavour quality. To obtain systems sensitive to lipid oxidation (LO), a formulation design was applied varying the composition of fatty matter and eggs. Oxidation of polyunsaturated fatty acids (PUFA) and formation of related volatile compounds were followed at the different steps of cake-making. Optimised dynamic Solid Phase Micro Extraction was applied to selectively extract either volatile or semi-volatile compounds directly from the baking vapours. We show for the first time that in the case of alveolar baked products, lipid oxidation occurs very early during the step of dough preparation and to a minor extent during the baking process. The generation of lipid oxidation compounds depends on PUFA content and on the presence of endogenous antioxidants in the raw matter. Egg yolk seemed to play a double role on reactivity: protecting unsaturated lipids from oxidation and being necessary to generate a broad class of compounds of the Maillard reaction during baking and linked to the typical flavour of sponge cake. Copyright © 2013 Elsevier Ltd. All rights reserved.

  15. A Biotin Switch-Based Proteomics Approach Identifies 14-3-3ζ as a Target of Sirt1 in the Metabolic Regulation of Caspase-2

    PubMed Central

    Andersen, Joshua L.; Thompson, J. Will; Lindblom, Kelly R.; Johnson, Erika S.; Yang, Chih-Sheng; Lilley, Lauren R.; Freel, Christopher D.; Moseley, M. Arthur; Kornbluth, Sally

    2011-01-01

    While lysine acetylation in the nucleus is well characterized, comparatively little is known about its significance in cytoplasmic signaling. Here we show that inhibition of the Sirt1 deacetylase, which is primarily cytoplasmic in cancer cell lines, sensitizes these cells to caspase-2-dependent death. To identify relevant Sirt1 substrates, we developed a novel proteomics strategy, enabling the identification of a range of putative substrates, including 14-3-3ζ, a known direct regulator of caspase-2. We show here that inhibition of Sirtuin activity accelerates caspase activation and overrides caspase-2 suppression by nutrient abundance. Furthermore, 14-3-3ζ is acetylated prior to caspase activation, and supplementation of Xenopus egg extract with glucose-6-phosphate, which promotes caspase-2/14-3-3ζ binding, enhances 14-3-3ζ-directed Sirtuin activity. Conversely, inhibiting Sirtuin activity promotes 14-3-3ζ dissociation from caspase-2 in both egg extract and human cell lines. These data reveal a role for Sirt1 in modulating apoptotic sensitivity, in response to metabolic changes, by antagonizing 14-3-3ζ acetylation. PMID:21884983

  16. Quantitative PCR and Digital PCR for Detection of Ascaris lumbricoides Eggs in Reclaimed Water

    PubMed Central

    Santísima-Trinidad, Ana Belén; Bornay-Llinares, Fernando Jorge; Martín González, Marcos; Pascual Valero, José Antonio; Ros Muñoz, Margarita

    2017-01-01

    The reuse of reclaimed water from wastewater depuration is a widespread and necessary practice in many areas around the world and must be accompanied by adequate and continuous quality control. Ascaris lumbricoides is one of the soil-transmitted helminths (STH) with risk for humans due to its high infectivity and an important determinant of transmission is the inadequacy of water supplies and sanitation. The World Health Organization (WHO) recommends a limit equal to or lower than one parasitic helminth egg per liter, to reuse reclaimed water for unrestricted irrigation. We present two new protocols of DNA extraction from large volumes of reclaimed water. Quantitative PCR (qPCR) and digital PCR (dPCR) were able to detect low amounts of A. lumbricoides eggs. By using the first extraction protocol, which processes 500 mL of reclaimed water, qPCR can detect DNA concentrations as low as one A. lumbricoides egg equivalent, while dPCR can detect DNA concentrations as low as five A. lumbricoides egg equivalents. By using the second protocol, which processes 10 L of reclaimed water, qPCR was able to detect DNA concentrations equivalent to 20 A. lumbricoides eggs. This fact indicated the importance of developing new methodologies to detect helminth eggs with higher sensitivity and precision avoiding possible human infection risks. PMID:28377928

  17. Loop-mediated isothermal amplification assay for detection and discrimination of Toxocara canis and Toxocara cati eggs directly from sand samples.

    PubMed

    Macuhova, K; Kumagai, T; Akao, N; Ohta, N

    2010-12-01

    We developed a novel and simple method, using loop-mediated isothermal amplification (LAMP), for the detection and discrimination of Toxocara canis and Toxocara cati eggs. The new method employs 4 steps: (1) concentration of Toxocara eggs in a small amount of sand; (2) dissolution of the proteinaceous membrane of eggs and simultaneously separation of them from the sand using NaClO treatment; (3) extraction of DNA using NaOH treatment; and (4) detection of T. canis / T. cati DNA using a LAMP assay. All these steps are fast, easy to perform, and do not require expensive equipment or reagents. The novel method was tested both experimentally and in a field study. In the laboratory, we could reliably detect as few as 3 T. canis eggs in artificially contaminated sand, if the experiment was repeated twice. In the field trial, we were able to detect T. cati DNA from 4 natural sandpits having moderate to heavy contamination, although not in a single lightly contaminated sandpit. All of the examined sandpits were found to be contaminated with eggs of T. cati, but none appeared to contain T. canis. Our new method could extract DNA from T. canis and T. cati eggs directly from sand samples as well as detect and distinguish these 2 species in a few easy steps, with markedly reduced time and expense.

  18. A rapid solid-phase extraction fluorometric method for thiamine and riboflavin in salmonid eggs

    USGS Publications Warehouse

    Zajicek, James L.; Tillitt, Donald E.; Brown, Scott B.; Brown, Lisa R.; Honeyfield, Dale C.; Fitzsimons, John D.

    2005-01-01

    A new method has been developed and successfully applied to the selective measurement of thiamine (nonphosphorylated), total thiamine (sum of thiamine, thiamine monophosphate [TMP], thiamine diphosphate [TDP], and thiamine triphosphate [TTP]), and potentially interfering riboflavin in acidic (2% trichloroacetic acid) extracts of selected salmonid and walleye egg samples. Acidic extracts of eggs were applied directly to end-capped C18, reversed-phase solid-phase extraction (SPE) columns and separated into three fractions by elution with mixtures of PO4 buffer (pH 2), methanol (10%), and acetonitrile (20%). All thiamine compounds recovered in the first two fractions were oxidized to their corresponding thiochromes with alkaline potassium hexacyanoferrate, and we measured the thiochrome fluorescence (excitation at 360 nm, emission at 460 nm) in a 96-well microplate reader. Riboflavin, recovered in third fraction (eluted with pH 2, 20% acetonitrile), was analyzed directly by measuring the fluorescence of this fraction (excitation at 450 nm, emission at 530 nm). Significant portions of the phosphate esters of thiamine (TMP, TDP, and presumably TTP), when present at low concentrations (< 10 nmol of total -thiamine per gram of egg), were not retained by the 100-mg SPE column, and were collected directly during sample loading and in a subsequent phosphoric acid rinse as fraction 1. Free thiamine (nonphosphorylated) and remaining portions of the TDP and TMP were then eluted in the second fraction with 10% methanol/PO4 buffer, whereas the un-ionized, relatively nonpolar riboflavin was eluted in the third fraction with 20% acetonitrile. This new method uses a traditional sample homogenization of egg tissue to extract thiamine compounds into 2% trichlororacetic acid solution; an inexpensive, commercially available SPE column; small amounts of sample (0.5-1 g); microliter volumes of solvents per sample; a traditional, relatively nonhazardous, oxidation of thiamine compounds to fluorescent thiochromes; and an ultraviolet-visible-wavelength-filter fluorometer for the measurements. ?? Copyright by the American Fisheries Society 2005.

  19. Mucolytic Activity Test of Shallot Extract (Allium Ascalonicum L) by in Vitro

    NASA Astrophysics Data System (ADS)

    Deswati, D. A.; Dhina, M. A.; Mubaroq, S. R.

    2018-01-01

    This paper aims to explain the results of research on the mucolytic activity of shallot extract is proportional to 0.2% N-Acetylcysteine. Shallot (Allium ascalonicum L.) is efficacious for treating cough. This research was conducted by examining the mucolytic activity of shallot extract made with various dose concentration 5%, 10%, 15%, 20%, and 25%. The mucolytic activity test was performed in vitro based on the decrease in the viscosity of the egg whites by using the Brookfield viscometer. The results showed that shallot extract with dose concentration of 5%, 10%, 15%, 20%, 25% had mucolytic activity by decreasing the viscosity of egg white solution. The effective concentration almost equal to 0.2% N-Acetylcysteine is at 25% concentration.

  20. Laboratory assessment of indigenous plant extracts for anti-juvenile hormone activity in Culex quinquefasciatus.

    PubMed

    Saxena, R C; Dixit, O P; Sukumaran, P

    1992-07-01

    Of 15 plants tested, five plant extracts showed anti-juvenile hormone-like activity against laboratory colonised late fourth instar larvae and adult female mosquitoes. Petroleum ether extract of Eichhornia crassipes and acetone extracts of Ageratum conyzoides, Cleome icosandra, Tagetes erectes and Tridax procumbens showed growth inhibitory (P less than 0.001) and juvenile hormone mimicing activity to the treated larvae of C. quinquefasciatus.. Larval pupal intermediates, demalanised pupae, defective egg rafts and adult with deformed flight muscles were few noticeable changes. Biting behaviour was observed to be affected only in Ageratum, Cleome and Tridax extracts (P less than 0.001). Loss of fecundity was observed in the treated mosquitoes but no sterilant effects could be seen. Adults, obtained from larvae exposed to the plant extracts produced significantly shorter egg-rafts (P less than 0.005) than in control.

  1. The integrin-binding motif RGDS induces protein tyrosine phosphorylation without activation in Bufo arenarum (Amphibia) oocytes.

    PubMed

    Mouguelar, Valeria S; Cabada, Marcelo O; Coux, Gabriela

    2011-05-01

    Integrins are cell adhesion molecules that are thought to be involved in sperm-oocyte interaction. Nevertheless, their function in mammalian fertilization is still controversial, as different species behave differently. In amphibians, their role is mainly supported by Xenopus laevis studies, where RGDS peptide induces oocyte activation. We recently provided evidence suggesting the presence and involvement of integrins in the interaction of the oocyte plasma membrane (PM) with sperm in the amphibian Bufo arenarum. In order to understand the role of integrin homologs in oocytes and their possible contribution to egg activation mechanisms, we examined the presence of integrin subunits and the effect of RGDS peptide on oocytes and during fertilization. Western blot studies detected integrin subunits α5, αV and β1 in oocytes. In sperm, we could detect only the αV integrin subunit. We found that RGDS peptide was unable to elicit egg activation or MAPK dephosphorylation, but can induce reversible inhibition of fertilization. A similar partial inhibition was produced by an anti-β1 integrin antibody. Using an anti-phosphotyrosine antibody we found major changes in phosphotyrosine-containing proteins in egg extracts minutes after fertilization. Cytosol and PMs isolated from oocytes and fertilized eggs showed additional fertilization-induced phosphorylated proteins. Some of these were also present in cytosol and PMs from RGDS-treated oocytes (partially mimicking fertilization). These findings suggest that B. arenarum fertilization involves integrins (e.g. β1 subunit) as adhesion proteins. Our data support the view that RGDS-binding receptors may function as signaling receptors in B. arenarum oocytes, but integrin engagement by RGDS is not sufficient for oocyte activation.

  2. Properties of low-fat, low-cholesterol egg yolk prepared by supercritical CO2 extraction.

    PubMed

    Bringe, N A

    1997-01-01

    A dry egg yolk ingredient called Eggcellent has 74% less fat and 90% less cholesterol than liquid egg yolks, when reconstituted on an equal protein basis. The phospholipids and proteins are retained, enabling the ingredient to have the taste and texturizing properties of fresh egg yolk. Using the new yolk, it is possible to significantly improve the acceptability of low-fat, low-cholesterol bakery products, scrambled eggs and mayonnaise dressings without losing nutritional claims. The structures and functional properties of egg yolk components and the conditions required to optimize their benefits in foods are reviewed. The lipoproteins of low-fat, low-cholesterol yolk have valuable properties as flavorants, texturizers, foaming agents, emulsifiers, antioxidants, colorants, and nutraceuticals.

  3. Oviposition traps to survey eggs of Lambdina fiscellaria (Lepidoptera: Geometridae).

    PubMed

    Hébert, Christian; Jobin, Luc; Auger, Michel; Dupont, Alain

    2003-06-01

    Outbreaks of the hemlock looper, Lambdina fiscellaria (Gueneé), are characterized by rapid increase and patchy distribution over widespread areas, which make it difficult to detect impending outbreaks. This is a major problem with this insect. Population forecasting is based on tedious and expensive egg surveys in which eggs are extracted from 1-m branches; careful observation is needed to avoid counting old unhatched eggs of previous year populations. The efficacy of artificial substrates as oviposition traps to sample hemlock looper eggs was tested as a means of improving outbreak detection and population forecasting. A white polyurethane foam substrate (1,095 lb/ft3) used with the Luminoc insect trap, a portable light trap, was highly efficient in sampling eggs of the hemlock looper. Foam strips placed on tree trunks at breast height were less efficient but easier and less expensive to use for the establishment of extensive survey networks. Estimates based on oviposition traps were highly correlated with those obtained from the 1-m branch extraction method. The oviposition trap is a standard, inexpensive, easy, and robust method that can be used by nonspecialists. This technique makes it possible to sample higher numbers of plots in widespread monitoring networks, which is crucial for improving the management of hemlock looper populations.

  4. Computer vision system for egg volume prediction using backpropagation neural network

    NASA Astrophysics Data System (ADS)

    Siswantoro, J.; Hilman, M. Y.; Widiasri, M.

    2017-11-01

    Volume is one of considered aspects in egg sorting process. A rapid and accurate volume measurement method is needed to develop an egg sorting system. Computer vision system (CVS) provides a promising solution for volume measurement problem. Artificial neural network (ANN) has been used to predict the volume of egg in several CVSs. However, volume prediction from ANN could have less accuracy due to inappropriate input features or inappropriate ANN structure. This paper proposes a CVS for predicting the volume of egg using ANN. The CVS acquired an image of egg from top view and then processed the image to extract its 1D and 2 D size features. The features were used as input for ANN in predicting the volume of egg. The experiment results show that the proposed CSV can predict the volume of egg with a good accuracy and less computation time.

  5. Potentiation of the antiinflammatory effect of Anacardium occidentale (Linn.) stem-bark aqueous extract by grapefruit juice.

    PubMed

    Ojewole, J A O

    2004-04-01

    In an attempt to scientifically appraise some of the ethnomedical uses of Anacardium occidentale Linn. (family: Anacardiaceae), the present study was undertaken to examine the antiinflammatory effect of the plant's stem-bark aqueous extract in rats. Young adult male Wistar rats weighing 250-300 g were used. The antiinflammatory effect of A. occidentale stem-bark aqueous extract alone and in combination with grapefruit (Citrus paradisi Macf.) juice was investigated on fresh egg albumin-induced rat paw edema. Like diclofenac (100 mg/kg p.o.), aqueous extract of A. occidentale stem-bark (800 mg/kg p.o.) produced time-related, sustained and significant reduction (p < 0.05-0.001) of the fresh egg albumin-induced acute inflammation of the rat hind paw. However, the antiinflammatory effect of the plant extract was found to be approximately 8-15 times less than that of diclofenac. Coadministration of grapefruit juice (5 ml/kg p.o.) with A. occidentale stem-bark aqueous extract (800 mg/kg p.o.) or diclofenac (100 mg/kg p.o.) significantly potentiated (p < 0.05-0.001) the antiinflammatory effects of the crude plant extract and diclofenac on fresh egg albumin-induced rat paw edema. Although A. occidentale stem-bark aqueous extract is less potent than diclofenac as an antiinflammatory agent, the results of this experimental animal study indicate that the plant extract possesses antiinflammatory activity, and thus lend pharmacological support to the folkloric use of the plant in the management and/or control of arthritis and other inflammatory conditions among the Yoruba-speaking people of western Nigeria.

  6. Development of an enzyme-linked immunosorbent assay for seven sulfonamide residues and investigation of matrix effects from different food samples.

    PubMed

    Zhang, Hongyan; Wang, Lei; Zhang, Yan; Fang, Guozhen; Zheng, Wenjie; Wang, Shuo

    2007-03-21

    Direct competitive enzyme-linked immunosorbent assays (ELISA) were developed to detect a broad range of sulfonamides in various matrices. Screening for this class of antibiotics in pig muscle, chicken muscle, fish, and egg extracts was accomplished by simple, rapid extraction methods carried out with only phosphate-buffered saline (PBS) buffer. Twenty milliliters of extract solution was added to 4 g of sample to extract the sulfonamide residues, and sample extracts diluted with assay buffer were directly analyzed by ELISA; matrix effects could be avoided with 1:5 dilution of pig muscle, chicken muscle, and egg extracts with PBS and 1:5 dilution of fish extract with 1% bovine serum albumin (BSA)-PBS. For liver sample, the extraction method was a little more complicated; 2 g of sample was added to 20 mL of ethanol, mixed, and then centrifuged. The solvent of 10 mL of the upper liquid was removed, and the residues were dissolved in 10 mL of PBS and then filtered; the filtrate was diluted two-fold with 0.5% BSA-PBS for ELISA. These common methods were able to detect seven sulfonamide residues such as sulfisozole, sulfathiazole, sufameter, sulfamethoxypyridazine, sulfapyridine, sulfamethizole, and sulfachlorpyridazine in pig muscle, liver, chicken muscle, egg, and fish. The assay's detection limits for these compounds were less than 100 microg kg-1. Various extraction methods were tested, and the average recovery (n=3) of 100 microg kg-1 for the matrices was found to range from 77.3 to 123.7%.

  7. In vitro anthelmintic activity of Combretum molle (R. Br. ex G. Don) (Combretaceae) against Haemonchus contortus ova and larvae.

    PubMed

    Ademola, I O; Eloff, J N

    2010-04-19

    Parasitic nematodes, especially Haemonchus contortus (Rudolphi), are among the most common and economically important causes of disease in sheep and goats owned by pastoralists and small holder farmers in Africa. The control of these infections relies mainly on the use of anthelmintic drugs. However, herbal preparations are widely used by pastoralists and small holder farmers for the treatment of their livestock against helminth parasites. The anthelmintic effect of acetone leaf extract and fractions of Combretum molle was investigated to determine the relative efficacy of the components against gastrointestinal sheep nematodes. The fractions were obtained by solvent:solvent extraction from the acetone extract. These were evaluated for nematocidal activity by means of an egg hatch (EHA) and larval a development and viability assay (LDVA) in vitro. The effect of the test extracts on the hatchability of eggs and development of first to third stage larvae and the survival rate of the third stage larvae. H. contortus, were used to determine the relative bioactivities. Best-fit LC(50) values were computed using global model of nonlinear regression curve-fitting. The extracts inhibited egg hatching and development of the larvae of H. contortus in a concentration-dependent manner. Best-fit LC(50) values for the egg hatch test were 0.866, 0.333, 0.833, 0.747, and 0.065mg/mL for acetone extract, n-butanol, hexane, chloroform, and 35% water in methanol fractions, respectively. The best-fit LC(50) values for the LDVA were 0.604, 0.362, 1.077, 0.131 and 0.318mg/mL for the acetone extract, butanol, hexane, chloroform, and 35% water in methanol fractions, respectively. In the EHA the 35% water in methanol fraction was significantly more active than all the other fractions (p<0.05); however the activity was not significantly different with the LDVA. C. molle leaf could find application in anthelmintic therapy in veterinary practice.

  8. Identification of proteins in renaissance paintings by proteomics.

    PubMed

    Tokarski, Caroline; Martin, Elisabeth; Rolando, Christian; Cren-Olivé, Cécile

    2006-03-01

    The presented work proposes a new methodology based on proteomics techniques to identify proteins in old art paintings. The main challenging tasks of this work were (i) to find appropriate conditions for extracting proteins from the binding media without protein hydrolysis in amino acids and (ii) to develop analytical methods adapted to the small sample quantity available. Starting from microsamples of painting models (ovalbumin-based, which is the major egg white protein, and egg-based paintings), multiple extraction solutions (HCl, HCOOH, NH3, NaOH) and conditions (ultrasonic bath, mortar and pestle, grinding resin) were evaluated. The best results were obtained using a commercial kit including a synthetic resin, mortar and pestle to grind the sample in an aqueous solution acidified with trifluoroacetic acid at 1% with additional multiple steps of ultrasonic baths. The resulting supernatant was analyzed by MALDI-TOF in linear mode to verify the efficiency of the extraction solution. An enzymatic hydrolysis step was also performed for protein identification; the peptide mixture was analyzed by nanoLC/nanoESI/Q-q-TOF MS/MS with an adapted chromatographic run for the low sample quantity. Finally, the developed methodology was successfully applied to Renaissance art painting microsamples of approximately 10 microg from Benedetto Bonfigli's triptych, The Virgin and Child, St. John the Baptist, St. Sebastian (XVth century), and Niccolo di Pietro Gerini's painting, The Virgin and Child (XIVth century), identifying, for the first time and without ambiguity, the presence of whole egg proteins (egg yolk and egg white) in a painting binder.

  9. Guar foaming albumin: a low molecular mass protein with high foaming activity and foam stability isolated from guar meal.

    PubMed

    Shimoyama, Ami; Kido, Shoko; Kinekawa, Yoh-ichi; Doi, Yukio

    2008-10-08

    The water extract of guar meal ( Cyamopsis tetragonolobus) was examined for its foamability. Compared with egg white, the extract showed an extraordinary foam stability: no drainage after 3 h of standing in contrast to 65% drainage for egg white at the same protein concentration. The acid-precipitated protein from the extract was responsible for the high foamability and designated guar foaming albumin (GFA). The foaming activity of GFA was 20 times higher than that of egg white. GFA consisted of two subunits with molecular masses of 6 and 11 kDa linked to each other through disulfide bonds. The cleavage of disulfide bonds in GFA affected the foamability only slightly. GFA remarkably decreased the surface tension of water at low protein concentrations. Immunoblotting analysis demonstrated that GFA did not react to the antisera from allergic patients against plant food. These results suggest that GFA serves as an effective food additive in developing protein-stabilized foam.

  10. Sage (Salvia officinalis) and fennel (Foeniculum vulgare) improve cryopreserved boar epididymal semen quality study.

    PubMed

    Monton, A; Gil, L; Malo, C; Olaciregui, M; Gonzalez, N; de Blas, I

    2015-01-01

    The aim of this study was to evaluate the effect of fennel and sage extracts and the influence of the egg yolk source (fresh or pasteurized) on the success of freezing boar epididymal spermatozoa. In experiment 1, epididymal sperm was recovered by flushing and cryopreserved in a lactose-egg yolk solution supplemented with various concentrations (10, 5 and 2.5 g/L) of sage or fennel. Sperm quality was evaluated (motility, viability, HOST and acrosome integrity) at 0 h and 2 h after thawing. Fennel 10 g/L and sage 5 g/L and control (no extracts) were selected for experiment 2 which also compared fresh or pasteurized egg yolk in the freezing extender and measured DNA integrity of the frozen sperm. Results showed that the interaction between fennel and sage antioxidants with fresh egg yolk significantly improved post thaw sperm quality and protected boar epididymal spermatozoa from cryopreservation damage as a result of oxidative stress.

  11. DNase I and II present in avian oocytes: a possible involvement in sperm degradation at polyspermic fertilisation.

    PubMed

    Stepińska, Urszula; Olszańska, Bozenna

    2003-02-01

    During polyspermic fertilisation in birds numerous spermatozoa enter the eggs, in contrast to the situation in mammals where fertilisation is monospermic. However, in birds only one of the spermatozoa which have entered an egg participates in zygote nucleus formation, while the supernumerary spermatozoa degenerate at early embryogenesis. Our previous work has demonstrated the presence in preovulatory quail oocytes of DNase I and II activities able to digest naked lambdaDNA/HindIII substrate in vitro. In the present studies, the activities of both DNases in quail oocytes at different stages of oogenesis and in ovulated mouse oocytes were assayed in vitro using the same substrate. Degradation of quail spermatozoa by quail oocyte extracts was also checked. Digestion of the DNA substrate was evaluated by electrophoresis on agarose gels. The activities of DNase I and II in quail oocytes increased during oogenesis and were the highest in mature oocytes. The activities were present not only in germinal discs but also in a thin layer of cytoplasm adhering to the perivitelline layer surrounding the yolk. At all stages of oogenesis the activity of DNase II was much higher than that of DNase I. DNA contained in spermatozoa was also degraded by the quail oocyte extracts under conditions optimal for both DNases. In contrast to what is observed in quail oocytes, no DNase activities were detected in ovulated mouse eggs; this is logical as they would be useless or even harmful in monospermic fertilisation. The possible role of DNase activities in avian oocytes, in degradation of accessory spermatozoa during polyspermic fertilisation, is discussed.

  12. Anthelminthic properties of Methylene chloride-methanol (1:1) extracts of two Cameroonians medicinal plants on Heligmosomoides bakeri (Nematoda: Heligmosomatidea).

    PubMed

    Ngouateu Teufack, Sergine Errole; NMbogning Tayo, Gertrude; Ngangout Alidou, Marc; Yondo, Jeannette; Djiomene, Amely Frankline; Wabo Poné, Josué; Mbida, Faùily Mpoame

    2017-08-11

    The resistance of some medico-veterinary parasite strains as well as the unavailability and toxicity of synthetic anthelminthics on humans, animals and the impacts of their residues in the environment have pushed scientists to turn to plants with anthelminthic properties. Hence, the aim of this work was to contribute to the fight against helminths of medical and veterinary importance in general, and also to clear the environment of their free living stages. Fresh eggs of Heligmosomoides bakeri were obtained from the faeces of experimentally infected mice. L 1 and L 2 larval stages were obtained after 48 and 72 h of coproculture respectively. Methylene Chloride-Methanol (1:1) extracts of Annona senegalensis and Nauclea latifolia were diluted in DMSO or Tween 80 to prepare the following concentrations: 625, 1250, 2500, 3750 and 5000 μg/ml. The effects of extract solutions were evaluated on the embryonation of eggs, egg hatching and on L 1 and L 2 survival after 48, 10 and 24 h of incubation. Negative controls were 1.5% DMSO, 4% Tween 80 and a mixture of these solvents. The TLC was carried out and the profiles of secondary metabolites were made. Negative controls had no effect on the embryonation, eggs hatching and on larval mortality. However, it was found that, the extracts affected the free living stages of H. bakeri in a concentration-dependant manner. At the highest concentration (5000 μg/ml), the rate of inhibition of embryonation obtained were 20.80%, 38.15% and 84.83% for Methylene Chloride-Methanol of Annona senegalensis (MCM As), Nauclea latifolia (MCM Nl) extracts and mixture of Annona senegalensis and Nauclea latifolia (MCM As-Nl) extract respectively. For egg hatch, the inhibition rate was 16.10%, 46.24% and 87.07% for the above three extracts respectively at the same concentration of 5000 μg/ml. On L 1 and L 2 larval stages after 24 h of exposure to extracts, the mortality rates of 100%, 54.76% and 96.77% against 98%, 51.44% and 100% were obtained for MCM As, MCM Nl and MCM As-Nl respectively at the highest concentration. The Methylene Chloride-Methanol of A.senegalensis, N. latifolia extracts showed the presence of alkaloids except in N. latifolia extract, flavonoids, sterols, triterpens, tanins, polyphenols, anthraquinons, saponins and terpenoids. These findings suggest that, the mixture of the two plant extracts showed an additive (synergetic effect) ovicidal effect and a slight larval mortality on L 1 as compared to the effect of MCM As extract alone. These effects were due to the presence ao secondary metabolites identifies in the plant extracts. Thus, they may be used as possible «disinfectants» for soil transmitted nematodes.

  13. Highly expressed amino acid biosynthesis genes revealed by global gene expression analysis of Salmonella enterica serovar Enteritidis during growth in whole egg are not essential for this growth.

    PubMed

    Jakočiūnė, Džiuginta; Herrero-Fresno, Ana; Jelsbak, Lotte; Olsen, John Elmerdahl

    2016-05-02

    Salmonella enterica serovar Enteritidis (S. Enteritidis) is the most common cause of egg borne salmonellosis in many parts of the world. This study analyzed gene expression of this bacterium during growth in whole egg, and whether highly expressed genes were essential for the growth. High quality RNA was extracted from S. Enteritidis using a modified RNA-extraction protocol. Global gene expression during growth in whole egg was compared to growth in LB-medium using DNA array method. Twenty-six genes were significantly upregulated during growth in egg; these belonged to amino acid biosynthesis, di/oligopeptide transport system, biotin synthesis, ferrous iron transport system, and type III secretion system. Significant downregulation of 15 genes related to formate hydrogenlyase (FHL) and trehalose metabolism was observed. The results suggested that S. Enteritidis is starved for amino-acids, biotin and iron when growing in egg. However, site specific mutation of amino acid biosynthesis genes asnA (17.3 fold upregulated), asnB (18.6 fold upregulated), asnA/asnB and, serA (12.0 fold upregulated) and gdhA (3.7 fold upregulated), did not result in growth attenuation, suggesting that biosynthesis using the enzymes encoded from these genes may represent the first choice for S. Enteritidis when growing in egg, but when absent, the bacterium could use alternative ways to obtain the amino acids. Copyright © 2016 Elsevier B.V. All rights reserved.

  14. Mosquito larvicidal, ovicidal, and repellent properties of botanical extracts against Anopheles stephensi, Aedes aegypti, and Culex quinquefasciatus (Diptera: Culicidae).

    PubMed

    Govindarajan, M; Mathivanan, T; Elumalai, K; Krishnappa, K; Anandan, A

    2011-08-01

    Mosquito-borne diseases have an economic impact, including loss in commercial and labor outputs, particularly in countries with tropical and subtropical climates; however, no part of the world is free from vector-borne diseases. In mosquito control programs, botanical origin may have the potential to be used successfully as eggs, larvae, and adult. The larvicidal, ovicidal, and repellent activities of crude benzene and ethyl acetate extracts of leaf of Ervatamia coronaria and Caesalpinia pulcherrima were assayed for their toxicity against three important vector mosquitoes, viz., Anopheles stephensi, Aedes aegypti, and Culex quinquefasciatus (Diptera: Culicidae). The larval mortality was observed after 24 h of exposure. All extracts showed moderate larvicidal effects; however, the highest larval mortality was found in benzene extract of E. coronaria against the larvae of Anopheles Stephensi, Aedes aegypti, and Culex quinquefasciatus with the LC(50) and LC(90) values were 79.08, 89.59, and 96.15 ppm and 150.47, 166.04, and 174.10 ppm, respectively. Mean percent hatchability of the ovicidal activity was observed 48 h posttreatment. The percent hatchability was inversely proportional to the concentration of extract and directly proportional to the eggs. The leaf extract of E. coronaria was found to be most effective than Caesalpinia pulcherrima against eggs/egg rafts of three vector mosquitoes. For E. coronaria, the benzene extract exerted 300, 250, and 200 ppm against Anopheles stephensi, Aedes aegypti, and Culex quinquefasciatus, respectively. The results of the repellent activity of benzene and ethyl acetate extract of E. coronaria and Caesalpinia pulcherrima plants at three different concentrations of 1.0, 2.5, and 5.0 mg/cm(2) were applied on skin of fore arm in man and exposed against adult female mosquitoes. In this observation, these two plant crude extracts gave protection against mosquito bites without any allergic reaction to the test person, and also, the repellent activity is dependent on the strength of the plant extracts. These results suggest that the leaf solvent plant extracts have the potential to be used as an ideal ecofriendly approach for the control of mosquitoes. This is the first report on the mosquito larvicidal, ovicidal, and repellent activities of the reported E. coronaria and Caesalpinia pulcherrima plants.

  15. 21 CFR 101.22 - Foods; labeling of spices, flavorings, colorings and chemical preservatives.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ..., leaf or similar plant material, meat, fish, poultry, eggs, dairy products, or fermentation products..., meat, seafood, poultry, eggs, dairy products, or fermentation products thereof, whose significant... does not include common salt, sugars, vinegars, spices, or oils extracted from spices, substances added...

  16. Physical properties evaluation of roselle extract-egg white mixture under various drying temperatures

    NASA Astrophysics Data System (ADS)

    Triyastuti, M. S.; Kumoro, A. C.; Djaeni, M.

    2017-03-01

    Roselle contains anthocyanin that is potential for food colorant. Occasionally, roselle extract is provided in dry powder prepared under high temperature. In this case, the anthocyanin color degrades due to the intervention of heat. The foammat drying with egg white is a potential method to speed up the drying process as well as minimize color degradation. This research aims to study the physical properties of roselle extract under foam mat drying. As indicators, the powder size and color intensity were observed. The result showed that at high temperatures, roselle powder under foam mat drying has the fine size with porous structure. However, at the higher the drying temperature the color retention decreased.

  17. Automatic stage identification of Drosophila egg chamber based on DAPI images

    PubMed Central

    Jia, Dongyu; Xu, Qiuping; Xie, Qian; Mio, Washington; Deng, Wu-Min

    2016-01-01

    The Drosophila egg chamber, whose development is divided into 14 stages, is a well-established model for developmental biology. However, visual stage determination can be a tedious, subjective and time-consuming task prone to errors. Our study presents an objective, reliable and repeatable automated method for quantifying cell features and classifying egg chamber stages based on DAPI images. The proposed approach is composed of two steps: 1) a feature extraction step and 2) a statistical modeling step. The egg chamber features used are egg chamber size, oocyte size, egg chamber ratio and distribution of follicle cells. Methods for determining the on-site of the polytene stage and centripetal migration are also discussed. The statistical model uses linear and ordinal regression to explore the stage-feature relationships and classify egg chamber stages. Combined with machine learning, our method has great potential to enable discovery of hidden developmental mechanisms. PMID:26732176

  18. In vitro effects of aqueous extract from Maytenus senegalensis (Lam.) Exell stem bark on egg hatching, larval migration and adult worms of Haemonchus contortus.

    PubMed

    Zangueu, Calvin Bogning; Olounlade, Abiodoun Pascal; Ossokomack, Marlyse; Djouatsa, Yolande Noelle Nangue; Alowanou, Goue Géorcelin; Azebaze, Anatole Guy Blaise; Llorent-Martínez, Eulogio José; de Córdova, Maria Luisa Fernández; Dongmo, Alain Bertrand; Hounzangbe-Adote, Mawulé Sylvie

    2018-05-02

    Maytenus senegalensis is a common shrub which is scattered in tropical Africa. Different parts of this plant have been reported to be useful in traditional medicine against gastrointestinal disorders and intestinal worms. This study evaluated the anthelmintic activity of the aqueous stem bark extract of M. senegalensis using egg hatch assay (EHA), larval migration inhibition assay (LMIA) and adult worms' motility inhibition assay (AMIA). On EHA, the extract concentrations tested resulted in a significant (p < 0.01) inhibition of egg hatching in concentration-dependent manner and ranged between 31.86% at 75 μg.mL - 1 to 54.92% at 2400 μg.mL - 1 after a 48 h post-exposure with eggs. For the LMI assays, the aqueous extract of M. senegalensis showed a significant (p < 0.05) inhibition of larval migration in a concentration-dependent manner. The highest concentration used (2400 μg.mL - 1 ) showed a 37.77% inhibition. The use of polyvinyl polypyrrolidone (PVPP) indicated that tannins and flavonoids were partly involved in the effect since the larval migration was inhibited by 15.5%, but other biochemical compounds were also implicated. On AMIA, M. senegalensis was associated with a reduced worm motility after a 24 h post exposure compared to phosphate buffered saline as control (p < 0.05). By this time 66.66% of the worms' were found immotile or dead in the wells containing plant extract at 2400 μg.mL - 1 . The Phytochemical analysis of aqueous extract of M. senegalensis by HPLC-ESI-MS n detected the presence of proanthocyanidins (20%) and flavonoids (> 50%). These in vitro results suggest the presence of some anthelmintic properties in M. senegalensis extract, which is traditionally used by small farmers in west and central Africa. These effects may be due to the flavonoids and proanthocyanidins present in the extract and need to be studied under in vivo conditions.

  19. Prevalence of internal parasites in beef cows in the United States: Results of the National Animal Health Monitoring System's (NAHMS) beef study, 2007-2008.

    PubMed

    Stromberg, Bert E; Gasbarre, Louis C; Ballweber, Lora R; Dargatz, David A; Rodriguez, Judith M; Kopral, Christine A; Zarlenga, Dante S

    2015-10-01

    During the United States Department of Agriculture (USDA) National Animal Health Monitoring System's (NAHMS) 2007-2008 beef study, 567 producers from 24 US States were offered the opportunity to collect fecal samples from weaned beef calves and have them evaluated for the presence of parasite eggs (Phase 1). Participating producers were provided with instructions and materials for sample collection. Up to 20 fresh fecal samples were collected from each of the 99 participating operations. Fresh fecal samples were submitted to one of 3 randomly assigned laboratories for evaluation. Upon arrival at the laboratories, all samples were processed for the enumeration of strongyle, Nematodirus, and Trichuris eggs using the modified Wisconsin technique. The presence or absence of coccidian oocysts and tapeworm eggs was also noted. In submissions where the strongyle eggs per gram exceeded 30, aliquots from 2 to 6 animals were pooled for DNA extraction. Extracted DNA was subjected to genus level polymerase chain reaction (PCR) identification for the presence of Ostertagia, Cooperia, Haemonchus, Oesophagostomum, and Trichostrongylus. In this study, 85.6% of the samples had strongyle type, Nematodirus, and Trichuris eggs. Among the samples evaluated, 91% had Cooperia, 79% Ostertagia, 53% Haemonchus, 38% Oesophagostomum, 18% Nematodirus, 7% Trichuris, and 3% Trichostrongylus. The prevalence of coccidia and tapeworm eggs was 59.9% and 13.7%, respectively.

  20. Prevalence of internal parasites in beef cows in the United States: Results of the National Animal Health Monitoring System’s (NAHMS) beef study, 2007–2008

    PubMed Central

    Stromberg, Bert E.; Gasbarre, Louis C.; Ballweber, Lora R.; Dargatz, David A.; Rodriguez, Judith M.; Kopral, Christine A.; Zarlenga, Dante S.

    2015-01-01

    During the United States Department of Agriculture (USDA) National Animal Health Monitoring System’s (NAHMS) 2007–2008 beef study, 567 producers from 24 US States were offered the opportunity to collect fecal samples from weaned beef calves and have them evaluated for the presence of parasite eggs (Phase 1). Participating producers were provided with instructions and materials for sample collection. Up to 20 fresh fecal samples were collected from each of the 99 participating operations. Fresh fecal samples were submitted to one of 3 randomly assigned laboratories for evaluation. Upon arrival at the laboratories, all samples were processed for the enumeration of strongyle, Nematodirus, and Trichuris eggs using the modified Wisconsin technique. The presence or absence of coccidian oocysts and tapeworm eggs was also noted. In submissions where the strongyle eggs per gram exceeded 30, aliquots from 2 to 6 animals were pooled for DNA extraction. Extracted DNA was subjected to genus level polymerase chain reaction (PCR) identification for the presence of Ostertagia, Cooperia, Haemonchus, Oesophagostomum, and Trichostrongylus. In this study, 85.6% of the samples had strongyle type, Nematodirus, and Trichuris eggs. Among the samples evaluated, 91% had Cooperia, 79% Ostertagia, 53% Haemonchus, 38% Oesophagostomum, 18% Nematodirus, 7% Trichuris, and 3% Trichostrongylus. The prevalence of coccidia and tapeworm eggs was 59.9% and 13.7%, respectively. PMID:26424909

  1. The influence of encapsulated embryos on the timing of hatching in the brooding gastropod Crepipatella dilatata

    NASA Astrophysics Data System (ADS)

    Andrade-Villagrán, P. V.; Baria, K. S.; Montory, J. A.; Pechenik, J. A.; Chaparro, O. R.

    2018-01-01

    Encapsulated embryos are generally thought to play an active role in escaping from egg capsules or egg masses. However, for species that brood their egg capsules, the factors controlling the timing of hatching are largely unclear, particularly the degree to which hatching is controlled by the embryos rather than by the mother, and the degree to which the hatching of one egg capsule influences the hatching of sister egg capsules within the same egg mass. We studied aspects of hatching using the direct-developing gastropod Crepipatella dilatata, which includes nurse eggs in its egg capsules and broods clusters of egg capsules for at least several weeks before metamorphosed juveniles are released. Isolated egg capsules were able to hatch successfully, in the absence of the mother. Moreover, the hatching of one capsule did not cause adjacent sister capsules to hatch. Hatched and un-hatched sister egg capsules from the same egg mass differed significantly in the number of metamorphosed juveniles, average shell size, offspring biomass (juveniles + veliger larvae), and the number of nurse eggs remaining per egg capsule. Differences in when egg capsules hatched within a single egg mass were not explained by differences in egg capsule age. Hatching occurred only after most nurse eggs had been ingested, most offspring had metamorphosed into juveniles, and juveniles had reached a mean shell length > 1.36 mm. Whether the mother has any role to play in coordinating the hatching process or juvenile release remains to be determined.

  2. Could the domestic cat play a significant role in the transmission of Echinococcus multilocularis? A study based on qPCR analysis of cat feces in a rural area in France

    PubMed Central

    Knapp, Jenny; Combes, Benoît; Umhang, Gérald; Aknouche, Soufiane; Millon, Laurence

    2016-01-01

    Echinococcus multilocularis, a cestode parasite responsible for alveolar echinococcosis in humans, is often reported in Europe. It involves red foxes, domestic dogs, and domestic and wild cats as definitive hosts. The parasite infects small mammals and accidentally humans as intermediate hosts and develops in a similar way to a tumor, usually in the liver. Domestic animals are suspected of playing a role in parasite transmission, but this is rarely proven. Moreover, the role of domestic cats is thought to be small, because of experimental studies showing incomplete development of the parasite observed in their intestines. In the present study, we investigated copro-sampling performed in a rural and highly endemic area in Eastern France, on carnivore feces (n = 150). From these samples, the parasite was detected and identified by DNA analysis using quantitative PCR targeting part of a mitochondrial gene (Em-qPCR). Taeniid eggs were isolated from positive-Em-qPCR samples by flotation, and species identification was confirmed by sequencing on DNA extracts. From a total of 43 copro-samples from cats, four tested positive for E. multilocularis by the Em-qPCR. In two of these, we found parasite eggs that were identified as E. multilocularis. This finding was confirmed by sequencing, while one dog stool out of 61 collected was found to be positive, no egg was detectable. At the same time, 34% of fox stools tested positive for the parasite. The present study challenges the current idea that cats are only of minor significance in the E. multilocularis life cycle. PMID:27739398

  3. Egg wash wastewater: estrogenic risk or environmental asset?

    PubMed

    Shappell, Nancy W

    2013-07-01

    Commercial production of eggs and egg products requires the washing of eggs to remove urinary-fecal material and broken egg residue. In the case of one Ohio farming facility, 1.6 million birds produce 1.4 million eggs per day, using approximately 50 mL of wash water/egg or approximately 70,000 L per day. The aqueous waste stream was evaluated for estrogenicity to determine if potential for endocrine disruption would result from agricultural application of such wastewater. Samples collected the Fall (October) of 2010 included: water from 2 egg washers operating in series, inlet pipe to the treatment lagoon, a lagoon composite, and products used within the facility in the cleaning of equipment and treatment of the waste. In February 2011, the treatment lagoon was fitted with an extensive aeration system and subsequent sample sets were collected on 3 consecutive days in May and November. Samples were extracted by solid phase extraction and assayed for estrogenic activity using the in vitro E-Screen assay. Raw untreated wastewater from the egg washers contained 17β-estradiol equivalents (E2 Eqs) ranging from 9 to 18 ng/L, pipe grab samples entering into the treatment lagoon ranged from <0.14 to 4.4 ng/L (variability related to time of emptying of egg wash tanks), whereas treatment lagoon water contained 0.3 to 4.0 ng/L E2 Eq. Addition of an aeration system to the treatment lagoon eliminated surface "frothing," reduced noxious odor emission, and E2 Eqs were lower than the pre-aeration concentrations (4 ng/L [n = 1, no statistical comparison possible] vs 0.3 to 1.4 ng/L in 2011). Because of matrix effects, estrogens were not quantifiable by LC-MS2 in even egg washwater extracts, at concentrations in which internal deuterated estrogen standards were quantifiable. Estrone and E2 parent ions were detected in egg washwater samples only, and confirmatory ion fragments were detected in only one of these samples. Estrogenicity of the wastewater from the treatment lagoon was already at the proposed aquatic no effect concentration for 17β-E2 and would be expected to decrease further as wastewater passes through 2 consecutive storage ponds before application on field crops for irrigation. The original project plan was to follow the wastewater as it was applied by aerial irrigation and concomitant surface runoff, but based on the consistent and extremely low concentration of estrogenic activity of the wastewater from the treatment lagoon, it was concluded that activity would be below limits of quantitation by E-Screen in water used for irrigation from the storage ponds. Use of egg wash wastewater--or gray water--to irrigate crops removes the cost and burden of wastewater treatment by the local wastewater plant, poses little to no potential threat of estrogenic endocrine disruption, and supports the conservation of water resources through the use of wastewater irrigation. Copyright © 2013 SETAC.

  4. Anthelmintic activity of extracts of Spondias mombin against gastrointestinal nematodes of sheep: studies in vitro and in vivo.

    PubMed

    Ademola, I O; Fagbemi, B O; Idowu, S O

    2005-04-01

    This study was carried out to validate the efficacy of Spondias mombin, used locally as an anthelmintic, and to standardize the effective dose of the plant extract required for worm control in livestock. In vitro and in vivo studies were conducted to determine the direct anthelmintic effect of ethanolic and aqueous extracts of S. mombin towards different ovine gastrointestinal nematodes. A larval development assay (LDA) was used to investigate the in vitro effect of extracts on strongyle larvae. Another study was conducted in vivo to evaluate the therapeutic efficacy of the extracts administered orally at dose rates of 125, 250, 500 mg/kg to sheep naturally infected with gastrointestinal nematodes. Twenty sheep were selected on the basis of positive faecal egg counts (750 epg). The sheep were allocated randomly to a non-medicated control group (A) or to groups given 125 mg/kg (B), 250 mg/kg (C) or 500 mg/kg (D) of extract, respectively. Sheep in groups B-D were given extracts orally on two days. Individual faecal egg counts were performed on days 0, 3, 6, 9 and 12. The presence of S. mombin extracts in in vitro cultures of larvae decreased the survival of L3 larvae. The LC50 of the aqueous extract of S. mombin was 0.907 mg/ml, while the LC50 of the ethanolic extract was 0.456 mg/ml. This difference in LC50 was statistically significant (p > 0.05). The mean percentage faecal egg reduction of sheep drenched with 500 mg/kg S. mombin extracts was 15.0%, 27.5%, 65.0%, 65.0%, 100.0% against Haenmonchus spp., Trichostrongylus spp., Oesophagostomunm spp., Strongyloides spp. and Trichuris spp. respectively, on day 12. Extracts of S. mombin could find application in the control of helminths in livestock.

  5. [Composition of chicken and quail eggs].

    PubMed

    Closa, S J; Marchesich, C; Cabrera, M; Morales, J C

    1999-06-01

    Qualified food composition data on lipids composition are needed to evaluate intakes as a risk factor in the development of heart disease. Proximal composition, cholesterol and fatty acid content of chicken and quail eggs, usually consumed or traded, were analysed. Proximal composition were determined using AOAC (1984) specific techniques; lipids were extracted by a Folch's modified technique and cholesterol and fatty acids were determined by gas chromatography. Results corroborate the stability of eggs composition. Cholesterol content of quail eggs is similar to chicken eggs, but it is almost the half content of data registered in Handbook 8. Differences may be attributed to the analytical methodology used to obtain them. This study provides data obtained with up-date analytical techniques and accessory information useful for food composition tables.

  6. Unveiling chemical defense in the rice stalk stink bug against the entomopathogenic fungus Metarhizium anisopliae.

    PubMed

    da Silva, Rodrigo Alves; Quintela, Eliane Dias; Mascarin, Gabriel Moura; Pedrini, Nicolás; Lião, Luciano Moraes; Ferri, Pedro Henrique

    2015-05-01

    Eggs, nymphs (1st-5th instar) and adults of Tibraca limbativentris were challenged by conidial suspensions of its major fungal pathogen Metarhizium anisopliae in order to assess their susceptibility. The role of chemical defensive compounds from exocrine secretions produced by both nymphs and adults were examined for their participation on M. anisopliae infection. Although insect susceptibility to M. anisopliae followed a dose-dependent manner, adults followed by older nymphs displayed the highest resistance. Eggs were highly susceptible showing >96% fungal infection. Crude extracts isolated from metathoracic scent gland and dorsal abdominal glands of adults and nymphs, respectively, showed fungistatic effects by impairing spore germination, vegetative growth and sporulation. Gas chromatography-mass spectrometry analysis of these extracts revealed that the major components were short-chain hydrocarbons (C10-13) and unsaturated aldehydes. In vitro tests with the corresponding synthetic standards indicated compounds with greater antifungal activity including (E)-2-hexenal, (E)-2-octenal, and (E)-2-decenal, with the latter being the most deleterious to fungal fitness. We demonstrated that differential susceptibility of the rice stalk stink bug to M. anisopliae infection is age-specific and partly mediated by fungistatic properties of aldehydes, which are produced by scent glands of both nymphs and adults. Copyright © 2015 Elsevier Inc. All rights reserved.

  7. Effects of Peganum harmala (Zygophyllaceae) seed extract on the olive fruit fly (Diptera: Tephritidae) and its larval parasitoid Psyttalia concolor (Hymenoptera: Braconidae).

    PubMed

    Rehman, Junaid Ur; Wang, Xin-Geng; Johnson, Marshall W; Daane, Kent M; Jilani, Ghulam; Khan, Mir A; Zalom, Frank G

    2009-12-01

    Peganum harmala L. (Zygophyllaceae) is an herb native to arid and semiarid regions of Central Asian deserts. This study investigated the effects of ethanol extracts of P. harmala seeds on the olive fruit fly, Bactrocera oleae (Rossi) (Diptera: Tephritidae), i.e., adult repellency, reproductive activity, and larval growth, as well as parasitism levels by Psyttalia concolor (Szépligeti). Olive fruit treated with 2% extract reduced B. oleae oviposition. In choice tests, female B. oleae spent >99% of their time foraging on untreated fruit rather than P. harmala-treated fruit. These changes in ovipositional behavior resulted in a nearly 30-fold decrease in oviposition marks on treated fruit compared with untreated fruit during a 48 h exposure period. When female B. oleae were fed liquid diet containing 0.2% P. harmala extract, there was no effect on the number of ovipositional marks on exposed fruit, but up to 21.4% of the deposited eggs were deformed. SDS-polyacrylamide gel electrophoresis analyses of deformed eggs revealed that some protein bands were missing. Consequently, the number of offspring produced by treated females was lower than by untreated females. Neither the sex ratio nor body size of the fly's offspring were affected by adults fed diet containing 0.2% P. harmala extract. However, there was a slightly prolonged developmental time from egg to adult. Parasitism of larval B. oleae by P. concolor was not affected by infested fruit treatment with 2% P. harmala extract. P. harmala extracts as a potential control for insect pest species are discussed.

  8. Optimization for quick, easy, cheap, effective, rugged and safe extraction of mycotoxins and veterinary drugs by response surface methodology for application to egg and milk.

    PubMed

    Zhou, Jian; Xu, Jiao-Jiao; Cong, Jin-Mi; Cai, Zeng-Xuan; Zhang, Jing-Shun; Wang, Jun-Lin; Ren, Yi-Ping

    2018-01-12

    A multiclass method was proposed for the simultaneous determination of various classes of veterinary drugs (n = 65), mycotoxins and metabolites (n = 39) in egg and milk by ultra-high performance liquid chromatography-tandem mass spectrometry. The contaminants were extracted by QuEChERS-based strategy including salt-out partitioning and dispersive solid-phase extraction for cleanup further. With the aim of maximizing throughput and extraction efficiency, Plackett-Burman design was employed initially for screening significant variables. And response surface methodology based on central composite design was conducted to achieve optimal conditions in details: 3.35% (v/v) of formic acid in acetonitrile, 1.2 g of NaCl, 0.5 g of anhydrous NaAc, 300 mg of C18 and 140 mg of primary secondary amine. Satisfactory analytical characteristics in validation, in aspects of accuracy (70%-105% for mycotoxins and quinolones, 55%-80% for sulphonamides and 40%-105% for other veterinary drugs), precision (inter-day RSDs < 14%) and sensitivity (LOQs ranged from 0.01 μg/kg to 31 μg/kg), were achieved under the optimized conditions. The matrix effects were evaluated and compensated by the use of matrix-matched calibration curves (R 2  > 0.987). In practice, 45 eggs and 30 milk samples were investigated by the established method, of which positive finding aflatoxin in milk and sterigmatocystin in eggs. Copyright © 2017 Elsevier B.V. All rights reserved.

  9. The Function of Anal Fin Egg-Spots in the Cichlid Fish Astatotilapia burtoni

    PubMed Central

    Theis, Anya; Salzburger, Walter; Egger, Bernd

    2012-01-01

    Color and pigmentation patterns of animals are often targets of sexual selection because of their role in communication. Although conspicuous male traits are typically implicated with intersexual selection, there are examples where sex-specific displays play a role in an intrasexual context, e.g. when they serve as signals for aggression level and/or status. Here, we focus on the function of a conspicuous male ornament in the most species-rich tribe of cichlid fishes, the haplochromines. A characteristic feature of these ca. 1500 species are so-called egg-spots in form of ovoid markings on the anal fins of males, which are made up of carotenoid based pigment cells. It has long been assumed that these yellow, orange or reddish egg-spots play an important role in the courtship and spawning behavior of these maternal mouth-brooding fishes by mimicking the eggs of a conspecific female. The exact function of egg-spots remains unknown, however, and there are several hypotheses about their mode of action. To uncover the function of this cichlid-specific male ornament, we used female mate choice experiments and a male aggression test in the haplochromine species Astatotilapia burtoni. We manipulated the number and arrangement of egg-spots on the anal fins of males, or removed them entirely, and tested (1) female preference with visual contact only using egg-traps, (2) female preference with free contact using paternity testing with microsatellites and (3) male aggression. We found that females did not prefer males with many egg-spots over males with fewer egg-spots and that females tended to prefer males without egg-spots over males with egg-spots. Importantly, males without egg-spots sired clutches with the same fertilization rate as males with egg-spots. In male aggression trials, however, males with fewer egg-spots received significantly more attacks, suggesting that egg-spots are an important signal in intrasexual communication. PMID:22242184

  10. Biological activity of Schinus molle on Triatoma infestans.

    PubMed

    Ferrero, A A; Werdin González, J O; Sánchez Chopa, C

    2006-07-01

    Hexanic extracts from leaves and fruits of Schinus molle were tested for repellent and insecticidal properties against first instar nymphs and eggs of Triatoma infestans, the vector of Chagas' disease. Leaf and fruit extracts were highly repellent for first nymphs. Fruit extracts had also ovicidal activity.

  11. Interaction of the nematophagous fungus Pochonia chlamydosporia and Parascaris equorum eggs in different culture media.

    PubMed

    de Carvalho, Lorendane Millena; Braga, Fabio Ribeiro; Domingues, Rafael Reis; Araujo, Juliana Milani; Lelis, Rosane Teixeira; de Paula, Alessandra Teixeira; da Silveira, Wendeo Ferreira; de Araújo, Jackson Victor

    2014-07-01

    Research involving the use of nematophagous fungi in the biological control of parasites of interest to veterinarians has occurred over recent years, with promising results. This article reports the infection of Parascaris equorum eggs by the fungus Pochonia chlamydosporia (isolates VC1 and VC4). Six groups were formed for each isolate, with six different culture media: 2% water-agar (2% WA); agar-chitin (AC); YPSSA (yeast extract, K2HPO4, MgSO4 ·7H2O, soluble starch); AELA extract (starch + water + agar); 2% corn-meal-agar (2% CMA); and 2% potato dextrose-agar (2% PDA). A total of 1000 eggs of P. equorum were transferred to each plate containing isolates grown for a period of 7 days (treatment group). Also, 1000 eggs were added to each plate without fungus (controlgroup). The plates were kept in an environmental chamber at 25 °C in the dark for 21 days. After, we analyzed the effects on ovicidal activity: effect 1 (accession shell); effect 2 (penetration hyphae); and effect 3 (destruction of the eggs). No differences were observed in the destruction of eggs between the two isolates. The decreasing effectiveness of the different culture media was: PDA (38.9%); CMA (38.3%); WA (36.7%); YPSSA (36.45%); and AC (32.5%). The highest percentage egg destruction was observed when the strains were grown in culture medium AELA (44.9%); this was the best medium. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  12. Isolation of Cholesterol from an Egg Yolk

    ERIC Educational Resources Information Center

    Taber, Douglass F.; Li, Rui; Anson, Cory M.

    2011-01-01

    A simple procedure for the isolation of the cholesterol, by hydrolysis and extraction followed by column chromatography, is described. The cholesterol can be further purified by complexation with oxalic acid. It can also be oxidized and conjugated to cholestenone. The source of the cholesterol is one egg yolk, which contains about 200 mg of…

  13. Phospholipids, Dietary Supplements, and Chicken Eggs: An Inquiry-Based Exercise Using Thin-Layer Chromatography

    ERIC Educational Resources Information Center

    Potteiger, Sara E.; Belanger, Julie M.

    2015-01-01

    This inquiry-based experiment is designed for organic or biochemistry undergraduate students to deduce the identity of phospholipids extracted from chicken eggs and dietary supplements. This is achieved using thin-layer chromatography (TLC) data, a series of guided questions of increasing complexity, and provided relative retention factor (Rf)…

  14. Egg serpins: The chicken and/or the egg dilemma.

    PubMed

    Dombre, Clara; Guyot, Nicolas; Moreau, Thierry; Monget, Philippe; Da Silva, Mylène; Gautron, Joël; Réhault-Godbert, Sophie

    2017-02-01

    Twenty-seven serpins belonging to clade A, B, C, D, E, F, G, H and I serpins are currently referenced in chicken genome databases. Phylogenetic analysis of chicken serpins revealed that ovalbumin (Serpinb14) and its paralogs ovalbumin-related protein Y (Serpinb14b) and ovalbumin-related protein X (Serpinb14c) are found in bird species. These clade B serpins are specifically expressed in reproductive tissues and exported in the egg where they constitute major protein components. These data suggest that these three paralogs have probably appeared in birds to face new environments and ensure the extra-uterine development of an embryo in a shell egg. Twelve other serpins have been identified in the newly produced egg, some of them having a specific distribution in the respective egg structures (eggshell, egg white, vitelline membrane and egg yolk). The physiological role of these egg serpins remain largely unexplored, but there is increasing evidence in literature or by homologies with their mammalian counterparts, that some of them participate in cell proliferation, tissue remodeling and/or angiogenesis associated with folliculogenesis and development of extraembryonic structures, eggshell biomineralization, egg defense and nutrition of the embryo. A better knowledge of the phylogenetic evolution of these 15 serpins in other oviparous species, on their egg distribution, on their regulation during embryonic development (activation/degradation/transfer) and on their functional specificity, is needed to better appreciate their role and their bird-specificity. These review shed light on the multiple possibilities that offer the avian egg model to study the role of serpins in reproduction and developmental biology. Copyright © 2016 The Author(s). Published by Elsevier Ltd.. All rights reserved.

  15. In vitro assessment of Argemone mexicana, Taraxacum officinale, Ruta chalepensis and Tagetes filifolia against Haemonchus contortus nematode eggs and infective (L3) larvae.

    PubMed

    Jasso Díaz, Gabriela; Hernández, Glafiro Torres; Zamilpa, Alejandro; Becerril Pérez, Carlos Miguel; Ramírez Bribiesca, J Efrén; Hernández Mendo, Omar; Sánchez Arroyo, Hussein; González Cortazar, Manasés; Mendoza de Gives, Pedro

    2017-08-01

    Argemone mexicana, Taraxacum officinale, Ruta chalepensis and Tagetes filifolia are plants with deworming potential. The purpose of this study was to evaluate methanolic extracts of aerial parts of these plants against Haemonchus contortus eggs and infective larvae (L3) and identify compounds responsible for the anthelmintic activity. In vitro probes were performed to identify the anthelmintic activity of plant extracts: egg hatching inhibition (EHI) and larvae mortality. Open column Chromatography was used to bio-guided fractionation of the extract, which shows the best anthelmintic effect. The lethal concentration to inhibit 50% of H. contortus egg hatching or larvae mortality (LC 50 ) was calculated using a Probit analysis. Bio-guided procedure led to the recognition of an active fraction (TF11) mainly composed by 1) quercetagitrin, 2) methyl chlorogenate and chlorogenic acid. Quercetagitrin (1) and methyl chlorogenate (2) did not show an important EHI activity (3-14%) (p < 0.05); however, chlorogenic acid (3) showed 100% of EHI (LC 50 248 μg/mL) (p < 0.05). Chlorogenic acid is responsible of the ovicidal activity and it seems that, this compound is reported for the first time with anthelmintic activity against a parasite of importance in sheep industry. Copyright © 2017 Elsevier Ltd. All rights reserved.

  16. Microwave assisted saponification (MAS) followed by on-line liquid chromatography (LC)-gas chromatography (GC) for high-throughput and high-sensitivity determination of mineral oil in different cereal-based foodstuffs.

    PubMed

    Moret, Sabrina; Scolaro, Marianna; Barp, Laura; Purcaro, Giorgia; Conte, Lanfranco S

    2016-04-01

    A high throughput, high-sensitivity procedure, involving simultaneous microwave-assisted extraction (MAS) and unsaponifiable extraction, followed by on-line liquid chromatography (LC)-gas chromatography (GC), has been optimised for rapid and efficient extraction and analytical determination of mineral oil saturated hydrocarbons (MOSH) and mineral oil aromatic hydrocarbons (MOAH) in cereal-based products of different composition. MAS has the advantage of eliminating fat before LC-GC analysis, allowing an increase in the amount of sample extract injected, and hence in sensitivity. The proposed method gave practically quantitative recoveries and good repeatability. Among the different cereal-based products analysed (dry semolina and egg pasta, bread, biscuits, and cakes), egg pasta packed in direct contact with recycled paperboard had on average the highest total MOSH level (15.9 mg kg(-1)), followed by cakes (10.4 mg kg(-1)) and bread (7.5 mg kg(-1)). About 50% of the pasta and bread samples and 20% of the biscuits and cake samples had detectable MOAH amounts. The highest concentrations were found in an egg pasta in direct contact with recycled paperboard (3.6 mg kg(-1)) and in a milk bread (3.6 mg kg(-1)). Copyright © 2015 Elsevier Ltd. All rights reserved.

  17. Oviposition Deterrence Induced by Ocimum kilimandscharicum and Ocimum suave Extracts to Gravid Anopheles gambiae s.s (Diptera: Culicidae) in Laboratory

    PubMed Central

    Kweka, Eliningaya J; Lyatuu, Ester E; Mboya, Michael A; Mwang’onde, Beda J; Mahande, Aneth M

    2010-01-01

    Background: In most of the past decades, mosquito control has been done by the use of indoor residual spray and insecticides-treated bed nets. The control of mosquitoes by targeting the breeding sites (larval habitat) has not been given priority. Disrupting the oviposition sensory detection of mosquitoes by introducing deterrents of plant origin, which are cheap resources, might be add value to integrated vector control. Such knowledge is required in order to successfully manipulate the behavior of mosquitoes for monitoring or control. Materials and Methods: Twenty gravid mosquitoes were placed in a cage measuring 30 × 30 × 30 cm for oviposition. The oviposition media were made of different materials. Experiments were set up at 6:00 pm, and eggs were collected for counting at 7:30 am. Mosquitoes were observed until they died. The comparisons of the number of eggs were made between the different treatments. Results: There was significant difference in the number of eggs found in control cups when compared with the number of eggs found in water treated with Ocimum kilimandscharicum (OK) (P=0.02) or Ocimum suave (OS) (P=0.000) and that found in water with debris treated with OK (P=0.011) or OS (P=0.002). There was no significant difference in the number of eggs laid in treated water and the number of eggs laid in water with debris treated either with OK (P=0.105) or OS (P=0.176). Oviposition activity index for both OS and OK experiments lay in a negative side and ranged from -0.19% to -1%. The results show that OS and OK deter oviposition in An.gambiae s.s. Conclusions: Further research needs to be done on the effect of secondary metabolites of these plant extracts as they decompose in the breeding sites. In the event of favorable results, the potential of these plant extracts can be harnessed on a larger scale. PMID:20927285

  18. Coplanar PCB distribution between chorioallantoic membranes and eggs of alligators and Loggerhead sea turtles

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Bargar, T.A.; Cobb, G.P.

    1995-12-31

    The relative distribution of coplanar polychlorinated biphenyls (PCBs) between chorioallantoic membranes (CAMS) and eggs was investigated in inviable American alligator (Alligator mississippiensis) and Loggerhead sea turtle (Caretta caretra) eggs. Cam and egg extracts were fractionated by HPLC using a porous graphitic column (PGC) and an in line switching valve to separate coplanar from non-coplanar PCBs. The fractions were collected, concentrated by nitrogen evaporation, and injected on GC-ECD (60M DB-5 capillary column) for quantification. Alligator and Loggerhead sea turtle eggs contain toxicologically significant coplanar PCBs. Mono-ortho substituted PCBs were present with greater frequency relative to non-ortho substituted PCBs in both eggsmore » and CAMS. The presence of coplanar PCBs in eggs appears to be correlated to coplanar PCB presence in CAMS. The chorioallantoic membrane could serve as a biomarker of embryo exposure to coplanar PCBs.« less

  19. Liquid chromatographic determination of 4,4'-dinitrocarbanilide, the active component of the infertility agent nicarbazin, in chicken, duck, goose, and snake eggs.

    PubMed

    Primus, Thomas M; Kohler, Dennis J; Goodall, Margaret A; Yoder, Christi; Mathies, Thomas; Miller, Lowell; Johnston, John J; Vercauteren, Kurt

    2003-01-01

    4,4'-Dinitrocarbanilide (DNC) was extracted from chicken, duck, goose, and snake eggs and isolated by reversed-phase liquid chromatography. DNC was detected by ultraviolet absorbance at 347 nm and quantitated by comparison with a calibration standard. Recoveries of DNC from fortified control chicken, duck, goose, and snake egg samples were determined for DNC levels of 0.16, 10, and 16 microg/g. The mean recoveries from chicken, duck, goose, and snake eggs were 92 +/- 4, 88 +/- 9, 87 +/- 7, and 95 +/- 6%, respectively. The method limits of detection for DNC in chicken, duck, goose, and snake eggs ranged from 0.015 to 0.035 microg/g. The reported method is much simpler than and equally efficient as previous methods developed for the determination of DNC residues in egg contents.

  20. Sperm and Spermatids Contain Different Proteins and Bind Distinct Egg Factors

    PubMed Central

    Teperek, Marta; Miyamoto, Kei; Simeone, Angela; Feret, Renata; Deery, Michael J.; Gurdon, John B.; Jullien, Jerome

    2014-01-01

    Spermatozoa are more efficient at supporting normal embryonic development than spermatids, their immature, immediate precursors. This suggests that the sperm acquires the ability to support embryonic development during spermiogenesis (spermatid to sperm maturation). Here, using Xenopus laevis as a model organism, we performed 2-D Fluorescence Difference Gel Electrophoresis (2D-DIGE) and mass spectrometry analysis of differentially expressed proteins between sperm and spermatids in order to identify factors that could be responsible for the efficiency of the sperm to support embryonic development. Furthermore, benefiting from the availability of egg extracts in Xenopus, we also tested whether the chromatin of sperm could attract different egg factors compared to the chromatin of spermatids. Our analysis identified: (1) several proteins which were present exclusively in sperm; but not in spermatid nuclei and (2) numerous egg proteins binding to the sperm (but not to the spermatid chromatin) after incubation in egg extracts. Amongst these factors we identified many chromatin-associated proteins and transcriptional repressors. Presence of transcriptional repressors binding specifically to sperm chromatin could suggest its preparation for the early embryonic cell cycles, during which no transcription is observed and suggests that sperm chromatin has a unique protein composition, which facilitates the recruitment of egg chromatin remodelling factors. It is therefore likely that the acquisition of these sperm-specific factors during spermiogenesis makes the sperm chromatin suitable to interact with the maternal factors and, as a consequence, to support efficient embryonic development. PMID:25244019

  1. Evolution of egg coats: linking molecular biology and ecology.

    PubMed

    Shu, Longfei; Suter, Marc J-F; Räsänen, Katja

    2015-08-01

    One central goal of evolutionary biology is to explain how biological diversity emerges and is maintained in nature. Given the complexity of the phenotype and the multifaceted nature of inheritance, modern evolutionary ecological studies rely heavily on the use of molecular tools. Here, we show how molecular tools help to gain insight into the role of egg coats (i.e. the extracellular structures surrounding eggs and embryos) in evolutionary diversification. Egg coats are maternally derived structures that have many biological functions from mediating fertilization to protecting the embryo from environmental hazards. They show great molecular, structural and functional diversity across species, but intraspecific variability and the role of ecology in egg coat evolution have largely been overlooked. Given that much of the variation that influences egg coat function is ultimately determined by their molecular phenotype, cutting-edge molecular tools (e.g. proteomics, glycomics and transcriptomics), combined with functional assays, are needed for rigorous inferences on their evolutionary ecology. Here, we identify key research areas and highlight emerging molecular techniques that can increase our understanding of the role of egg coats in the evolution of biological diversity, from adaptation to speciation. © 2015 John Wiley & Sons Ltd.

  2. Improved method for the on-line metal chelate affinity chromatography-high-performance liquid chromatographic determination of tetracycline antibiotics in animal products.

    PubMed

    Cooper, A D; Stubbings, G W; Kelly, M; Tarbin, J A; Farrington, W H; Shearer, G

    1998-07-03

    An improved on-line metal chelate affinity chromatography-high-performance liquid chromatography (MCAC-HPLC) method for the determination of tetracycline antibiotics in animal tissues and egg has been developed. Extraction was carried out with ethyl acetate. The extract was then evaporated to dryness and reconstituted in methanol prior to on-line MCAC clean-up and HPLC-UV determination. Recoveries of tetracycline, oxytetracycline, demeclocycline and chlortetracycline in the range 42% to 101% were obtained from egg, poultry, fish and venison tissues spiked at 25 micrograms kg-1. Limits of detection less than 10 microgram kg-1 were estimated for all four analytes. This method has higher throughput, higher recovery and lower limits of detection than a previously reported on-line MCAC-HPLC method which involved aqueous extraction and solid-phase extraction clean-up.

  3. A highly efficient, cell-free translation/translocation system prepared from Xenopus eggs.

    PubMed Central

    Matthews, G; Colman, A

    1991-01-01

    We describe the use of a Xenopus laevis egg extract for the in vitro translation and post translational modification of membrane and secretory proteins. This extract is capable of the translation and segregation into membranes of microgram per millilitre levels of protein from added mRNAs. Signal sequences of segregated proteins are efficiently cleaved and appropriate N-linked glycosylation patterns are produced. The extract also supports the quantitative assembly of murine immunoglobulin heavy and light chains into tetramers, and two events which take place beyond the endoplasmic reticulum, mannose 6 phosphorylation of murine cathepsin D and O-linked glycosylation of coronavirus E1 protein, also occur, but at reduced efficiency. The stability of the membranes allows protease protection studies and quantitative centrifugal fractionation of segregated and unsegregated proteins to be performed. Conditions for the use of stored extract have also been determined. Images PMID:1754376

  4. DOE Office of Scientific and Technical Information (OSTI.GOV)

    Swindle, T.D.; Caffee, M.W.; Hohenberg, C.M.

    The iodine-xenon systems of six Allende inclusions (five Eggs and the Pink Angel) appear to have been altered by non-nebular secondary processes. Evidence for this includes temperature-ordered variations in the initial I isotopic composition within several objects (with older apparent I-Xe ages associated with higher extraction temperatures) and the absence of primitive I-Xe ages. The span of apparent ages seen in Allende objects (10 Ma or more) is probably too long to reflect any nebular process, so at least some alteration probably occurred on the parent body. The range in initial {sup 244}Pu/{sup 238}U ratios for the Eggs (3-14 {times}more » 10{sup minus 3}) includes the current best estimates of the bulk solar system value (4-7 {times} 10{sup minus 3}). For Egg 3, the Pu/U ratio varies by a factor of two between extractions, probably the result of fractionation of Pu from U among different phases.« less

  5. Antioxidant and hepatoprotective effects of silymarin phytosomes compared to milk thistle extract in CCl4 induced hepatotoxicity in rats.

    PubMed

    El-Gazayerly, O N; Makhlouf, A I A; Soelm, A M A; Mohmoud, M A

    2014-01-01

    Milk thistle extract is a well-known hepatoprotectant with low bioavailability (20-50%). The objective of the present study is to prepare and characterize silymarin phytosomes and to test the hepatoprotective effect of the phytosomes in CCl4 induced liver injury in rats compared to milk thistle extract. Phytosomes were prepared using lecithin from soybeans and from egg yolk. The prepared phytosomes were examined using scanning electron microscopy, transmission electron microscopy, differential scanning calorimetry, Fourier transform infrared spectroscopy and proton nuclear magnetic resonance spectroscopy (H(1)NMR). The loading efficiency was >85% in all phytosomal formulations. Formula P2 (with the molar ratio of soybean lecithin to silybin 1:1) and P4 (with the molar ratio of egg-yolk lecithin to silybin 0.25:1) exhibited significantly (p < 0.05) faster release than milk thistle extract. The in vivo study revealed that phytosomes significantly (p < 0.05) decreased glutamic pyruvic transaminase and super oxide dismutase activities compared to milk thistle extract.

  6. Detection of Helminth Eggs and Identification of Hookworm Species in Stray Cats, Dogs and Soil from Klang Valley, Malaysia

    PubMed Central

    Mahmud, Rohela; Samsudin, Nur Izyan; Kek Heng, Chua; Ling, Lau Yee

    2015-01-01

    The present study was conducted to determine the prevalence of helminth eggs excreted in the faeces of stray cats, dogs and in soil samples. A total of 505 fresh samples of faeces (from 227 dogs and 152 cats) and soil were collected. The egg stage was detected via microscopy after the application of formalin–ether concentration technique. Genomic DNA was extracted from the samples containing hookworm eggs and used for further identification to the species level using real-time polymerase chain reaction coupled with high resolution melting analysis. Microscopic observation showed that the overall prevalence of helminth eggs among stray cats and dogs was 75.7% (95% CI = 71.2%–79.9%), in which 87.7% of dogs and 57.9% of cats were infected with at least one parasite genus. Five genera of heliminth eggs were detected in the faecal samples, including hookworms (46.4%), Toxocara (11.1%), Trichuris (8.4%), Spirometra (7.4%) and Ascaris (2.4%). The prevalence of helminth infections among stray dogs was significantly higher than that among stray cats (p < 0.001). Only three genera of helminths were detected in soil samples with the prevalence of 23% (95% CI = 15.1%–31%), consisting of hookworms (16.6%), Ascaris (4%) and Toxocara (2.4%). The molecular identification of hookworm species revealed that Ancylostoma ceylanicum was dominant in both faecal and soil samples. The dog hookworm, Ancylostoma caninum, was also detected among cats, which is the first such occurrence reported in Malaysia till date. This finding indicated that there was a cross-infection of A. caninum between stray cats and dogs because of their coexistent within human communities. Taken together, these data suggest the potential role of stray cats and dogs as being the main sources of environmental contamination as well as for human infections. PMID:26671680

  7. Measurement of Iron in Egg Yolk: An Instrumental Analysis Experiment Using Biochemical Principles

    ERIC Educational Resources Information Center

    Maloney, Kevin M.; Quiazon, Emmanuel M.; Indralingam, Ramee

    2008-01-01

    The generally accepted method to determine iron content in food is by acid digestion or dry ashing and subsequent flame atomic absorption spectrometry or inductively coupled plasma atomic emission spectrometry. We have developed an experiment that chemically extracts the iron from an egg yolk and quantifies it using UV-vis absorption…

  8. Characterization of Tolypocladium cylindrosporum (Hypocreales: Ophiocordycipitacea) and its impact against Aedes aegypti and Aedes albopictus eggs at low temperature

    USDA-ARS?s Scientific Manuscript database

    We examined the growth characteristics of Tolypocladium cylindrosporum IBT 41712 and its potential to infect eggs of Aedes aegypti Linnaeus and Aedes albopictus (Skuse) mosquitoes at low temperature (15 deg C). When grown on sabouraud dextrose agar supplemented with yeast extract, IBT 41712 formed w...

  9. Morinda citrifolia L. leaf extract as antibacterial Salmonella typhimurium to increase productivity of quail (Coturnix coturnix japonica).

    PubMed

    Retnani, Y; Dan, T M Wardiny; Taryati

    2014-04-01

    The objective of this study was to apply effect of Morinda citrifolia L. citrifolia L. leaf extract as antibacterial of Salmonella typhimurium on mortality of Day Old Quail (DOQ), egg production and Hen day, hatchability of layer quail. This research was conducted at Laboratory of microbiology and laboratory of poultry nutrition, faculty of animal science, bogor agricultural university and slamet quail farms cilangkap, sukabumi, west java, Indonesia on March-July 2012. Two hundred and forty heads of quail were randomly assigned to four dietary treatments (sixty heads of quail/treatment). Experimental design used was Completely Randomized Design (CRD). The treatments consist of level of biscuit Morinda citrifolia L. Citrifolia L. leaf extract i.e R1 = 0%, R2 = 5%, R3 = 10%, R4 = 15%. The results indicated the treatments had significant effect (p < 0.05) on mortality of Day Old Quail (DOQ). The average mortality of Day Old Quail (DOQ) was given extract Morinda citrifolia L. leaf were R1 (4.00%), R2 (1.00%), R3 (1.33%), R4 (0.67%). The average mortality of Day Old Quail (DOQ) was given 15% extract Morinda citrifolia L. leaf (R4) was lowest than control treatment (R1). The results of the analysis indicated that Morinda citrifolia L. leaf of quail drink had not significant effect (p > 0.05) on egg production, hen day and hatchability. It was concluded that the Morinda citrifolia L. leaf extract 15% can reduce mortality of Day Old Quail (DOQ) and can increase its egg production, hen day and hatchability.

  10. A Teaching Laboratory for Comprehensive Lipid Characterization from Food Samples

    ERIC Educational Resources Information Center

    Bendinskas, Kestutis; Weber, Benjamin; Nsouli, Tamara; Nguyen, Hoangvy V.; Joyce, Carolyn; Niri, Vadoud; Jaskolla, Thorsten W.

    2014-01-01

    Traditional and state-of-the-art techniques were combined to probe for various lipid classes from egg yolk and avocado qualitatively and quantitatively. A total lipid extract was isolated using liquid-liquid extraction. An aliquot of the total lipid extract was subjected to transesterification to form volatile fatty acid methyl esters suitable for…

  11. A deep learning framework to discern and count microscopic nematode eggs.

    PubMed

    Akintayo, Adedotun; Tylka, Gregory L; Singh, Asheesh K; Ganapathysubramanian, Baskar; Singh, Arti; Sarkar, Soumik

    2018-06-14

    In order to identify and control the menace of destructive pests via microscopic image-based identification state-of-the art deep learning architecture is demonstrated on the parasitic worm, the soybean cyst nematode (SCN), Heterodera glycines. Soybean yield loss is negatively correlated with the density of SCN eggs that are present in the soil. While there has been progress in automating extraction of egg-filled cysts and eggs from soil samples counting SCN eggs obtained from soil samples using computer vision techniques has proven to be an extremely difficult challenge. Here we show that a deep learning architecture developed for rare object identification in clutter-filled images can identify and count the SCN eggs. The architecture is trained with expert-labeled data to effectively build a machine learning model for quantifying SCN eggs via microscopic image analysis. We show dramatic improvements in the quantification time of eggs while maintaining human-level accuracy and avoiding inter-rater and intra-rater variabilities. The nematode eggs are correctly identified even in complex, debris-filled images that are often difficult for experts to identify quickly. Our results illustrate the remarkable promise of applying deep learning approaches to phenotyping for pest assessment and management.

  12. Cytotoxic, analgesic and anti-inflammatory properties of Syzygium calophyllifolium bark.

    PubMed

    Chandran, Rahul; Abrahamse, Heidi; Parimelazhagan, Thangaraj

    2018-04-25

    The present study explores the effect of Syzygium calophyllifolium bark methanol (SCBM) extract on pain and inflammation as cancer associated symptoms. The anti-proliferative and cell death-inducing ability of the extract was analysed using MCF-7 breast cancer cells. ATP and LDH levels, along with the cell morphology were noted. The anti-proliferative ability of the extract was examined in relation to pain and inflammation. Analgesic activity was determined using a hot plate, acetic acid and formalin-induced pain models in mice. Acute anti-inflammatory activity was observed in carrageenan and egg albumin induced paw oedema in Wistar rats. Chronic inflammation was induced by placing a cotton pellet under the skin at the axial region of rats. Reduced ATP and increased LDH content indicated the cytotoxic effect of the extract. SCBM at 200 mg/kg dose depicted good activity in acetic acid (77.69%) and formalin (80.81%) induced pain models compared to the standard drug indomethacin. However, only a moderate activity was observed in the hot plate method (53.85%). The higher dose of SCBM was equally effective in anti-inflammatory models. The dose (200 mg/kg) significantly reduced the paw oedema in carrageenan (96.71%) and egg albumin models (54.24%) compared to the control. The chronic inflammation was also inhibited upto 70.46% by SCBM. These activities can be attributed to the phenolic compounds detected in HPLC. The outcome of the study states that the extract can reduce both acute and chronic inflammatory conditions and kill the cancer cells, proposing the dual role of SCBM in treating inflammatory pains in cancer. The result of these studies indicated that wild plants like Syzygium calophyllifolium could be taken forward for a detailed study to accomplish the demand for a better treatment against diseases. Copyright © 2018 Elsevier Masson SAS. All rights reserved.

  13. Avian Egg and Egg Coat.

    PubMed

    Okumura, Hiroki

    2017-01-01

    An ovulated egg of vertebrates is surrounded by unique extracellular matrix, the egg coat or zona pellucida, playing important roles in fertilization and early development. The vertebrate egg coat is composed of two to six zona pellucida (ZP) glycoproteins that are characterized by the evolutionarily conserved ZP-domain module and classified into six subfamilies based on phylogenetic analyses. Interestingly, investigations of biochemical and functional features of the ZP glycoproteins show that the roles of each ZP-glycoprotein family member in the egg-coat formation and the egg-sperm interactions seemingly vary across vertebrates. This might be one reason why comprehensive understandings of the molecular basis of either architecture or physiological functions of egg coat still remain elusive despite more than 3 decades of intensive investigations. In this chapter, an overview of avian egg focusing on the oogenesis are provided in the first section, and unique features of avian egg coat, i.e., perivitelline layer, including the morphology, biogenesis pathway, and physiological functions are discussed mainly on chicken and quail in terms of the characteristics of ZP glycoproteins in the following sections. In addition, these features of avian egg coat are compared to mammalian zona pellucida, from the viewpoint that the structural and functional varieties of ZP glycoproteins might be associated with the evolutionary adaptation to their reproductive strategies. By comparing the egg coat of birds and mammals whose reproductive strategies are largely different, new insights into the molecular mechanisms of vertebrate egg-sperm interactions might be provided.

  14. In vitro and in vivo anthelmintic activity of crude extracts of Coriandrum sativum against Haemonchus contortus.

    PubMed

    Eguale, T; Tilahun, G; Debella, A; Feleke, A; Makonnen, E

    2007-04-04

    In vitro anthelmintic activities of crude aqueous and hydro-alcoholic extracts of the seeds of Coriandrum sativum (Apiaceae) were investigated on the egg and adult nematode parasite Haemonchus contortus. The aqueous extract of Coriandrum sativum was also investigated for in vivo anthelmintic activity in sheep infected with Haemonchus contortus. Both extract types of Coriandrum sativum inhibited hatching of eggs completely at a concentration less than 0.5 mg/ml. ED(50) of aqueous extract of Coriandrum sativum was 0.12 mg/ml while that of hydro-alcoholic extract was 0.18 mg/ml. There was no statistically significant difference between aqueous and hydro-alcoholic extracts (p>0.05). The hydro-alcoholic extract showed better in vitro activity against adult parasites than the aqueous one. For the in vivo study, 24 sheep artificially infected with Haemonchus contortus were randomly divided into four groups of six animals each. The first two groups were treated with crude aqueous extract of Coriandrum sativum at 0.45 and 0.9 g/kg dose levels, the third group with albendazole at 3.8 mg/kg and the last group was left untreated. Efficacy was tested by faecal egg count reduction (FECR) and total worm count reduction (TWCR). On day 2 post treatment, significant FECR was detected in groups treated with higher dose of Coriandrum sativum (p<0.05) and albendazole (p<0.001). On days 7 and 14 post treatment, significant FECR was not detected for both doses of Coriandrum sativum (p>0.05). Significant (p<0.05) TWCR was detected only for higher dose of Coriandrum sativum compared to the untreated group. Reduction in male worms was higher than female worms. Treatment with both doses of Coriandrum sativum did not help the animals improve or maintain their PCV while those treated with albendazole showed significant increase in PCV (p<0.05).

  15. Development and optimization of a naphthoic acid-based ionic liquid as a "non-organic solvent microextraction" for the determination of tetracycline antibiotics in milk and chicken eggs.

    PubMed

    Gao, Jiajia; Wang, Hui; Qu, Jingang; Wang, Huili; Wang, Xuedong

    2017-01-15

    In traditional ionic liquids (ILs)-based microextraction, ILs are often used as extraction and dispersive solvents; however, their functional effects are not fully utilized. Herein, we developed a novel ionic liquid 1-butyl-3-methylimidazolium naphthoic acid salt ([C4MIM][NPA]) with strong acidity. It was used as a mixed dispersive solvent with conventional [C2MIM][BF4] in "functionalized ionic liquid-based non-organic solvent microextraction (FIL-NOSM)" for determination of tetracycline antibiotics (TCs) in milk and eggs. Utilization of [C4MIM][NPA] in FIL-NOSM method increased extraction recoveries (ERs) of TCs by more than 20% and eliminated the pH adjustment step because of its strong acidity. Under optimized conditions based on central composite design, the ERs of four TCs were 94.1-102.1%, and the limitsofdetection were 0.08-1.12μgkg(-1) in milk and egg samples. This proposed method provides high extraction efficiency, less pretreatment time and requires non-organic solvents for determination of trace TC concentrations in complex animal-based food matrices. Copyright © 2016 Elsevier Ltd. All rights reserved.

  16. EVALUATIN OF ANTIHYPERTENSIVE ACTIVITY OF SONCHUS ASPER L. IN RATS.

    PubMed

    Mushtaq, Muhammad Naveed; Akhtar, Muhammad Shoaib; Alamgeer; Ahmad, Taseer; Khan, Hafeez Ullah; Maheen, Safirah; Ahsan, Haseeb; Naz, Huma; Asif, Hira; Younis, Waqas; Tabassum, Nazia

    2016-01-01

    The present investigation was carried out to evaluate the effect of aerial parts of Sonchus asper L. in normotensive, glucose and egg feed diet induced hypertensive rats. Aqueous-methanolic extract of Sonchus asper in 250, 500 and 1000 mg/kg doses was studied in normotensive and glucose induced hypertensive rats using the non-invasive technique. The results obtained showed that the extract has significantly (p < 0.5 - p < 0.001) decreased the blood pressure and heart rate in dose dependent manner. The dose 1000 mg/kg of the extract produced the maximum antihypertensive effect and was selected for further experiments. The extract was found to prevent the rise in blood pressure of egg and glucose fed rats as compared to control group in 21 days study. The LD50 of the plant extract was 3500 mg/kg b.w. in mice and sub-chronic toxicity study showed that there was no significant alteration in the blood chemistry of the extract treated rats. It is conceivable, therefore, that the aqueous-methanolic extract of Sonchus asper has exerted considerable antihypertensive activity in rats and has duly supported traditional medicinal use of plant in hypertension.

  17. Short-term effects of lower oil dried distillers grains with solubles in laying hen rations.

    PubMed

    Purdum, Sheila; Hanford, Kathy; Kreifels, Brett

    2014-10-01

    Extraction of oil from dried distillers grains has become a common practice among US ethanol producers. The valuable oil has been diverted to markets other than poultry feed, leaving new dried distillers grains with solubles (DDGS) products higher in fiber and purportedly lower in ME. This study compared 3 DDGS products with 10.3, 7.3, or 5.2% ether extract, respectively, with a corn-soy control ration in young Bovan laying hens for a feeding period from 20 to 33 wk of age. The DDGS was fed at the rate of 20% of the ration. Lower oil content of DDGS had no effect on short-term egg production parameters: feed intake, egg production, egg weight or mass, and hen weight gain. The diets containing lower fat DDGS (5.2%) did have reduced AME and kilocalories per day intake for laying hens. For each percent reduction in oil from a normal DDGS sample (10.3%) to medium oil (7.3%) DDGS, AME decreased 42.3 kcal/kg of diet. However, total kilocalories per day intake did sustain good egg production during this short trial. ©2014 Poultry Science Association Inc.

  18. Effects of toxic compounds in Montipora capitata on exogenous and endogenous zooxanthellae performance and fertilization success.

    PubMed

    Hagedorn, Mary; Farrell, Ann; Carter, Virginia; Zuchowicz, Nikolas; Johnston, Erika; Padilla-Gamiño, Jacqueline; Gunasekera, Sarath; Paul, Valerie

    2015-01-01

    Studies have identified chemicals within the stony coral genus Montipora that have significant biological activities. For example, Montiporic acids A and B and other compounds have been isolated from the adult tissue and eggs of Montipora spp. and have displayed antimicrobial activity and cytotoxicity in cultured cells. The ecological role of these toxic compounds is currently unclear. This study examines the role these toxins play in reproduction. Toxins were found in the eggs and larvae of the coral Montipora capitata. Releasing these toxins by crushing both the eggs and larvae resulted in irreversible inhibition of photosynthesis in endogenous and exogenous zooxanthellae within minutes. Moreover, these toxins were stable, as frozen storage of eggs and larvae did not affect toxicity. Photosynthetic competency of Porites compressa zooxanthellae treated with either frozen or fresh, crushed eggs was inhibited similarly (P > 0.05, ANCOVA). Addition of toxic eggs plugs to live P. compressa fragments caused complete tissue necrosis under the exposed area on the fragments within 1 week. Small volumes of M. capitata crushed eggs added to sperm suspensions reduced in vitro fertilization success by killing the sperm. After 30 min, untreated sperm maintained 90 ± 1.9% SEM motility while those treated with crushed eggs were rendered immotile, 4 ± 1.4% SEM. Flow cytometry indicated membrane disruption of the immotile sperm. Fertilization success using untreated sperm was 79 ± 4% SEM, whereas the success rate dropped significantly after exposure to the crushed eggs, 1.3 ± 0% SEM. Unlike the eggs and the larvae, M. capitata sperm did not reduce the photosynthetic competency of P. compressa zooxanthellae, suggesting the sperm was nontoxic. The identity of the toxins, cellular mechanism of action, advantage of the toxins for M. capitata and their role on the reef are still unknown.

  19. Egg yolk antibodies for detection and neutralization of Clostridium botulinum type A neurotoxin.

    PubMed

    Trott, D L; Yang, M; Gonzalez, J; Larson, A E; Tepp, W H; Johnson, E A; Cook, M E

    2009-05-01

    The objective of this research project was to determine the usefulness of an egg antibody platform for producing materials for the detection and neutralization of botulinum type A neurotoxin. Yield estimates for detection and neutralizing antibodies produced using methods described were calculated. Antibody specific to botulinum toxoid A (aToxoid) and toxin A (aBoNT/A) was produced by immunizing hens with botulinum toxoid A (toxoid) followed by increasing amounts of botulinum neurotoxin A (BoNT/A) in Freund incomplete adjuvant. Egg yolks were extracted with polyethylene glycol (PEG) for antibody detection and neutralization experiments. A model aToxoid/toxoid immunoassay using only egg yolk antibody was developed and had a detection limit of 1 pg/ml of toxoid. In an indirect enzyme-linked immunosorbent assay of BoNT/A-specific antibody, the aBoNT/A contained more BoNT/A-specific antibody than did the aToxoid, and aBoNT/A was as effective as commercial rabbit antibody. The aToxoid provided no protection against BoNT/A in a standard mouse neutralization assay; however, 1 mg of PEG-extracted aBoNT/A neutralized 4,000 lethal doses of BoNT/A injected intraperitoneally. Based on these results, we calculated that in 1 month one hen could produce more than 100 liters of antibody detection reagents or enough antibody to neutralize approximately 11.6 million mouse lethal doses of botulinum toxin. Utilization of an egg antibody platform is potentially rapid (28 to 70 days) and scalable to kilogram quantities using current egg production facilities with as few as 1,000 hens.

  20. Identification of double-yolked duck egg using computer vision.

    PubMed

    Ma, Long; Sun, Ke; Tu, Kang; Pan, Leiqing; Zhang, Wei

    2017-01-01

    The double-yolked (DY) egg is quite popular in some Asian countries because it is considered as a sign of good luck, however, the double yolk is one of the reasons why these eggs fail to hatch. The usage of automatic methods for identifying DY eggs can increase the efficiency in the poultry industry by decreasing egg loss during incubation or improving sale proceeds. In this study, two methods for DY duck egg identification were developed by using computer vision technology. Transmittance images of DY and single-yolked (SY) duck eggs were acquired by a CCD camera to identify them according to their shape features. The Fisher's linear discriminant (FLD) model equipped with a set of normalized Fourier descriptors (NFDs) extracted from the acquired images and the convolutional neural network (CNN) model using primary preprocessed images were built to recognize duck egg yolk types. The classification accuracies of the FLD model for SY and DY eggs were 100% and 93.2% respectively, while the classification accuracies of the CNN model for SY and DY eggs were 98% and 98.8% respectively. The CNN-based algorithm took about 0.12 s to recognize one sample image, which was slightly faster than the FLD-based (about 0.20 s). Finally, this work compared two classification methods and provided the better method for DY egg identification.

  1. Identification of double-yolked duck egg using computer vision

    PubMed Central

    Ma, Long; Sun, Ke; Tu, Kang; Pan, Leiqing; Zhang, Wei

    2017-01-01

    The double-yolked (DY) egg is quite popular in some Asian countries because it is considered as a sign of good luck, however, the double yolk is one of the reasons why these eggs fail to hatch. The usage of automatic methods for identifying DY eggs can increase the efficiency in the poultry industry by decreasing egg loss during incubation or improving sale proceeds. In this study, two methods for DY duck egg identification were developed by using computer vision technology. Transmittance images of DY and single-yolked (SY) duck eggs were acquired by a CCD camera to identify them according to their shape features. The Fisher’s linear discriminant (FLD) model equipped with a set of normalized Fourier descriptors (NFDs) extracted from the acquired images and the convolutional neural network (CNN) model using primary preprocessed images were built to recognize duck egg yolk types. The classification accuracies of the FLD model for SY and DY eggs were 100% and 93.2% respectively, while the classification accuracies of the CNN model for SY and DY eggs were 98% and 98.8% respectively. The CNN-based algorithm took about 0.12 s to recognize one sample image, which was slightly faster than the FLD-based (about 0.20 s). Finally, this work compared two classification methods and provided the better method for DY egg identification. PMID:29267387

  2. Control of Haemonchus contortus in sheep using basidiocarps of Agaricus blazei Murril.

    PubMed

    Vieira, Thallyta Maria; Fonseca, Leydiana Duarte; Bastos, Gabriela Almeida; de Oliveira Vasconcelos, Viviane; Silva, Maria Luíza França; Morais-Costa, Franciellen; de Paiva Ferreira, Adriano Vinícius; Oliveira, Neide Judith Faria de; Duarte, Eduardo Robson

    2017-06-01

    This study evaluated the effects in vitro and in vivo of Agaricus blazei against Haemonchus contortus in sheep. The in vitro efficacy of aqueous extract on egg hatching inhibition (EHI) was investigated and after 72 h incubation with varying concentrations the effects on, blastomeres, embryonated eggs, and first stage larvae (L1) were evaluated. Larval development inhibition (LDI) for dry powder and the aqueous extract were evaluated in fecal cultures of sheep infected with H. contortus. In vivo efficacy was determined by reduction in fecal egg count (FEC). Lambs were treated with powder A. blazei (11.4 g/kg pc) or trichlorfon, or were untreated and the possible toxicity of this fungus was monitored by plasmatic enzyme analysis. Concentrations equal to and higher than 3.62 mg/mL and of aqueous extract were 100% effective in the EHI test. In the LDI test, LC90 was estimated for 5.66 and 106.0 mg/g fecal culture for aqueous extract and powder, respectively. The mean FEC in lambs 14 days post-treatment with A. blazei powder was significantly lower than observed for the negative control, and the serum levels of aspartate transaminase and alanine transaminase were normal. The fungi supplementation promotes, respectively, high and moderate anthelmintic efficacy with in vitro and in vivo tests, respectively, suggesting it as an alternative or complementary treatment for haemonchosis in sheep.

  3. Calculating the Degradation Rate of Individual Proteins Using Xenopus Extract Systems.

    PubMed

    McDowell, Gary S; Philpott, Anna

    2018-05-16

    The Xenopus extract system has been used extensively as a simple, quick, and robust method for assessing the stability of proteins against proteasomal degradation. In this protocol, methods are provided for assessing the half-life of in vitro translated radiolabeled proteins using Xenopus egg or embryo extracts. © 2019 Cold Spring Harbor Laboratory Press.

  4. Effects of xylem-sap composition on glassy-winged sharpshooter egg maturation on high and low quality host plants

    USDA-ARS?s Scientific Manuscript database

    Glassy-winged sharpshooters must feed as adults to produce mature eggs. Cowpea and sunflower are both readily accepted by the glassy-winged sharpshooter for feeding, but egg production on sunflower was reported to be lower than egg production on cowpea. To better understand the role of adult diet in...

  5. A nonlethal microsampling technique to monitor the effects of mercury on wild bird eggs

    USGS Publications Warehouse

    Stebbins, Katherine R.; Klimstra, Jon D.; ,; Ackerman, Joshua T.; Heinz, Gary

    2009-01-01

    Methylmercury is the predominant chemical form of mercury reported in the eggs of wild birds, and the embryo is the most sensitive life stage to methylmercury toxicity. Protective guidelines have been based mainly on captive-breeding studies with chickens (Gallus gallus), mallards (Anas platyrhynchos), and ring-necked pheasants (Phasianus colchicus) or on field studies where whole eggs were collected and analyzed and the effects of the mercury were measured based on the reproductive success of the remaining eggs. However, both of these methods have limitations. As an alternative, we developed a technique that involves extracting a small sample of albumen from a live egg, sealing the egg, returning the egg to its nest to be naturally incubated by the parents, and then relating the hatching success of this microsampled egg to its mercury concentration. After first developing this technique in the laboratory using chicken and mallard eggs, we selected the laughing gull (Larus atricilla) and black-necked stilt (Himantopus mexicanus) as test subjects in the field. We found that 92% of the microsampled laughing gull eggs met our reproductive endpoint of survival to the beginning of hatching compared to 100% for the paired control eggs within the same nests. Microsampled black-necked stilt eggs exhibited 100% hatching success compared to 93% for the paired control eggs. Our results indicate that microsampling is an effective tool for nonlethally sampling mercury concentrations in eggs and, as such, can be used for monitoring sensitive species, as well as for improving studies that examine the effects of mercury on avian reproduction.

  6. Rapid Ovary Mass-Isolation (ROMi) to Obtain Large Quantities of Drosophila Egg Chambers for Fluorescent In Situ Hybridization.

    PubMed

    Jambor, Helena; Mejstrik, Pavel; Tomancak, Pavel

    2016-01-01

    Isolation of large quantities of tissue from organisms is essential for many techniques such as genome-wide screens and biochemistry. However, obtaining large quantities of tissues or cells is often the rate-limiting step when working in vivo. Here, we present a rapid method that allows the isolation of intact, single egg chambers at various developmental stages from ovaries of adult female Drosophila flies. The isolated egg chambers are amenable for a variety of procedures such as fluorescent in situ hybridization, RNA isolation, extract preparation, or immunostaining. Isolation of egg chambers from adult flies can be completed in 5 min and results, depending on the input amount of flies, in several milliliters of material. The isolated egg chambers are then further processed depending on the exact requirements of the subsequent application. We describe high-throughput in situ hybridization in 96-well plates as example application for the mass-isolated egg chambers.

  7. Leisingera sp. JC1, a Bacterial Isolate from Hawaiian Bobtail Squid Eggs, Produces Indigoidine and Differentially Inhibits Vibrios

    PubMed Central

    Gromek, Samantha M.; Suria, Andrea M.; Fullmer, Matthew S.; Garcia, Jillian L.; Gogarten, Johann Peter; Nyholm, Spencer V.; Balunas, Marcy J.

    2016-01-01

    Female members of many cephalopod species house a bacterial consortium in the accessory nidamental gland (ANG), part of the reproductive system. These bacteria are deposited into eggs that are then laid in the environment where they must develop unprotected from predation, pathogens, and fouling. In this study, we characterized the genome and secondary metabolite production of Leisingera sp. JC1, a member of the roseobacter clade (Rhodobacteraceae) of Alphaproteobacteria isolated from the jelly coat of eggs from the Hawaiian bobtail squid, Euprymna scolopes. Whole genome sequencing and MLSA analysis revealed that Leisingera sp. JC1 falls within a group of roseobacters associated with squid ANGs. Genome and biochemical analyses revealed the potential for and production of a number of secondary metabolites, including siderophores and acyl-homoserine lactones involved with quorum sensing. The complete biosynthetic gene cluster for the pigment indigoidine was detected in the genome and mass spectrometry confirmed the production of this compound. Furthermore, we investigated the production of indigoidine under co-culture conditions with Vibrio fischeri, the light organ symbiont of E. scolopes, and with other vibrios. Finally, both Leisingera sp. JC1 and secondary metabolite extracts of this strain had differential antimicrobial activity against a number of marine vibrios, suggesting that Leisingera sp. JC1 may play a role in host defense against other marine bacteria either in the eggs and/or ANG. These data also suggest that indigoidine may be partially, but not wholly, responsible for the antimicrobial activity of this squid-associated bacterium. PMID:27660622

  8. The function of Xenopus Bloom's syndrome protein homolog (xBLM) in DNA replication

    PubMed Central

    Liao, Shuren; Graham, Jeanine; Yan, Hong

    2000-01-01

    The Bloom's syndrome gene (BLM) plays a pivotal role in the maintenance of genomic stability in somatic cells. It encodes a DNA helicase (BLM) of the RecQ family, but the exact function of BLM remains elusive. To study this question, we have cloned the BLM homolog of the frog Xenopus laevis (xBLM) and have raised antibodies to it. Immunodepletion of xBLM from a Xenopus egg extract severely inhibits the replication of DNA in reconstituted nuclei. Moreover, the inhibition can be rescued by the addition of the recombinant xBLM protein. These results provide the first direct evidence that BLM plays an important role in DNA replication, suggesting that Bloom's syndrome may be the consequence of defective DNA replication. PMID:11040210

  9. Antioxidant activity in cooked and simulated digested eggs.

    PubMed

    Remanan, M K; Wu, J

    2014-07-25

    The avian egg is an excellent source of nutrients consisting of components with beneficial properties but there is limited knowledge on the effect of cooking methods and gastrointestinal digestion on the antioxidant activity of eggs. The present study was focused on the effect of cooking and simulated gastrointestinal digestion on antioxidant activity of eggs using ORAC, ABTS and DPPH assays. The results suggest that fresh egg yolk has higher antioxidant activity than fresh egg white and whole eggs. Cooking reduced whereas simulated gastrointestinal digestion increased the antioxidant activity of eggs. Boiled egg white hydrolysate showed the highest antioxidant activity; a total of 63 peptides were identified, indicative of the formation of novel antioxidant peptides upon simulated gastrointestinal digestion. This study suggests the potential role of eggs as a dietary source of antioxidants.

  10. Microbiology and Safety of Table Eggs

    USDA-ARS?s Scientific Manuscript database

    This chapter describes the microbiology of table eggs, effects of processing, regulatory influences, relative risk of egg-borne disease, and the role of retail and consumer practices in outbreaks. Effects of washing, refrigeration, and facility sanitation in US commercial facilities will be describe...

  11. Effects of Aqueous Extracts of Copaifera langsdorffii (Fabaceae) on the Growth and Reproduction of Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctuidae).

    PubMed

    Sâmia, R R; de Oliveira, R L; Moscardini, V F; Carvalho, G A

    2016-10-01

    Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctuidae) is considered a pest of maize crops throughout the Western Hemisphere. We report on the effects of aqueous extracts of leaves and bark of Copaifera langsdorffii (Fabaceae) on the biology of S. frugiperda, as an alternative source of natural bioactive molecules for the sustainable management of this pest. Second instars were sprayed with aqueous extracts prepared with 5% (w/v) plant material and/or fed on an artificial diet containing extracts at a concentration equivalent to 0.25% (w/v) for 17 days. Both leaf and bark extracts of C. langsdorffii significantly reduced S. frugiperda food intake, feces, and larval weight and caused a delay in larval development. Additionally, C. langsdorffii-based extracts increased the oviposition period; induced morphological changes in the eggs, including deformation of the corium and malformation of the micropylar and aeropylar regions; and reduced egg viability. But, aqueous extracts of C. langsdorffii exhibited no negative effects on larval and pupal survival, duration of the pupal stage, survival of pupae, sex ratio, longevity, duration of pre-oviposition period, and female fecundity. Aqueous extracts of leaves and bark of C. langsdorffii are promising alternatives for the control of S. frugiperda in maize crops.

  12. Determination of carotenoids and all-trans-retinol in fish eggs by liquid chromatography-electrospray ionization-tandem mass spectrometry.

    PubMed

    Li, Hongxia; Tyndale, Sélène T; Heath, Daniel D; Letcher, Robert J

    2005-02-25

    A novel method was developed for the combined determination of carotenoids and retinoids in fish eggs, which incorporates prior analyte isolation using liquid-liquid partitioning to minimize analyte degradation, and fraction analysis using high-performance liquid chromatography-electrospray (positive)-quadrupole mass spectrometry (LC-ESI(+)-MS; SIM or MRM modes). Eggs from Chinook salmon (Oncorhynchus tshawytscha) were used as the model fish egg matrix. The methodology was assessed and validated for beta-carotene, lutein, zeaxanthin, and beta-cryptoxanthin (molecular ion radicals [M](+)), canthaxanthin and astaxanthin ([M+Na](+) adducts) and all-trans-retinol ([(M+H)-H(2)O](+)). Using replicate egg samples (n=5) spiked with beta-cryptoxanthin and beta-carotene before and after extraction, matrix-sourced ESI(+) enhancement was observed as evidenced by comparable %matrix effect and %process efficiency values for beta-cryptoxanthin and beta-carotene of 114-119%. In aquaculture-raised eggs from adult Chinook salmon astaxanthin, all-trans-retinol, lutein and canthaxanthin were identified and determined at concentrations of 4.12, 1.06, 0.12 and 0.45 microg/g (egg wet weight), respectively. To our knowledge, this is the first report on a method for LC-MS determination of carotenoids and retinoids in a fish egg matrix, and the first carotenoid-specific determination in any fish egg sample.

  13. Nup53 Is Required for Nuclear Envelope and Nuclear Pore Complex Assembly

    PubMed Central

    Hawryluk-Gara, Lisa A.; Platani, Melpomeni; Santarella, Rachel

    2008-01-01

    Transport across the nuclear envelope (NE) is mediated by nuclear pore complexes (NPCs). These structures are composed of various subcomplexes of proteins that are each present in multiple copies and together establish the eightfold symmetry of the NPC. One evolutionarily conserved subcomplex of the NPC contains the nucleoporins Nup53 and Nup155. Using truncation analysis, we have defined regions of Nup53 that bind to neighboring nucleoporins as well as those domains that target Nup53 to the NPC in vivo. Using this information, we investigated the role of Nup53 in NE and NPC assembly using Xenopus egg extracts. We show that both events require Nup53. Importantly, the analysis of Nup53 fragments revealed that the assembly activity of Nup53 depleted extracts could be reconstituted using a region of Nup53 that binds specifically to its interacting partner Nup155. On the basis of these results, we propose that the formation of a Nup53–Nup155 complex plays a critical role in the processes of NPC and NE assembly. PMID:18256286

  14. Assessing the potential for egg chemoattractants to mediate sexual selection in a broadcast spawning marine invertebrate

    PubMed Central

    Evans, Jonathan P.; Garcia-Gonzalez, Francisco; Almbro, Maria; Robinson, Oscar; Fitzpatrick, John L.

    2012-01-01

    In numerous species, egg chemoattractants play a critical role in guiding sperm towards unfertilized eggs (sperm chemotaxis). Until now, the known functions of sperm chemotaxis include increasing the effective target size of eggs, thereby promoting sperm–egg encounters, and facilitating species recognition. Here, we report that in the broadcast spawning mussel, Mytilus galloprovincialis, egg chemoattractants may play an unforeseen role in sexual selection by enabling sperm to effectively ‘choose’ between the eggs of different conspecific females. In an initial experiment, we confirmed that sperm chemotaxis occurs in M. galloprovincialis by showing that sperm are attracted towards unfertilized eggs when given the choice of eggs or no eggs in a dichotomous chamber. We then conducted two cross-classified mating experiments, each comprising the same individual males and females crossed in identical male × female combinations, but under experimental conditions that offered sperm ‘no-choice’ (each fertilization trial took place in a Petri dish and involved a single male and female) or a ‘choice’ of a female's eggs (sperm were placed in the centre of a dichotomous choice chamber and allowed to choose eggs from different females). We show that male-by-female interactions characterized fertilization rates in both experiments, and that there was remarkable consistency between patterns of sperm migration in the egg-choice experiment and fertilization rates in the no-choice experiment. Thus, sperm appear to exploit chemical cues to preferentially swim towards eggs with which they are most compatible during direct sperm-to-egg encounters. These results reveal that sperm differentially select eggs on the basis of chemical cues, thus exposing the potential for egg chemoattractants to mediate mate choice for genetically compatible partners. Given the prevalence of sperm chemotaxis across diverse taxa, our findings may have broad implications for sexual selection in other mating systems. PMID:22438495

  15. Determination of egg proteins in snack food and noodles.

    PubMed

    Williams, Kristina M; Westphal, Carmen D; Shriver-Lake, Lisa C

    2004-01-01

    Egg is one of the 5 major allergenic foods that are responsible for more than 3/4 of food allergies in children. Food-allergic responses can be controlled by avoidance of the offending foods. The applicability of a commercial enzyme-linked immunosorbent assay (ELISA) kit for the detection of egg in food products such as cookies, crackers, pretzels, salad dressings, and raw and cooked noodles was evaluated. A preliminary evaluation of an antibody-based biosensor was also performed. A National Institute of Standards and Technology (NIST) whole dried egg powder reference material, SRM 8415, was used as a standard. A homogeneous and stable aqueous egg suspension was prepared for the evaluation of the performance of the Veratox for Egg Allergen Test (Neogen Corp., Lansing, MI). This test does not detect egg yolk proteins. Each gram of the aqueous dried egg suspension contained 643 microg whole dried egg, 0.5 mg thimerosal, and 2.5 mg bovine serum albumin. When cookies, crackers, salad dressings, noodles, and ice cream were spiked at a level of 24 mg/kg SRM 8415, recoveries for whole egg averaged about 28%. All foods containing egg as indicated on the ingredient label were found positive by the Veratox test. No false positives occurred in samples that did not contain eggs. Similar results were obtained using the Naval Research Laboratory (NRL) array biosensor, an evanescent wave fluoroimmunosensor. Results for cooked noodles showed that they contained <1% of the egg found in uncooked noodles. A comparison of extracts from cooked and uncooked noodles by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) revealed differences in protein profiles. The boiling of the noodles could have reduced the immunoreactivity of the egg proteins to the antibodies used in the kit or rendered the egg proteins nonextractable.

  16. Microneedle-based analysis of the micromechanics of the metaphase spindle assembled in Xenopus laevis egg extracts

    PubMed Central

    Shimamoto, Yuta; Kapoor, Tarun M.

    2014-01-01

    SUMMARY To explain how micron-sized cellular structures generate and respond to forces we need to characterize their micromechanical properties. Here we provide a protocol to build and use a dual force-calibrated microneedle-based set-up to quantitatively analyze the micromechanics of a metaphase spindle assembled in Xenopus laevis egg extracts. This cell-free extract system allows for controlled biochemical perturbations of spindle components. We describe how the microneedles are prepared and how they can be used to apply and measure forces. A multi-mode imaging system allows tracking of microtubules, chromosomes and needle tips. This set-up can be used to analyze the viscoelastic properties of the spindle on time-scales ranging from minutes to sub-seconds. A typical experiment, along with data analysis, is also detailed. We anticipate that our protocol can be readily extended to analyze the micromechanics of other cellular structures assembled in cell-free extracts. The entire procedure can take 3-4 days. PMID:22538847

  17. Potential miRNA regulators of differential HPG axis gene expression between low egg producing and high egg producing turkey hens

    USDA-ARS?s Scientific Manuscript database

    Expression differences exist in key genes of the hypothalamo-pituitary-gonadal (HPG) axis in low egg producing hens (LEPH) and high egg producing hens (HEPH); however, regulation of these differences is unknown. MicroRNAs (miRNAs) are small non-coding RNAs that play a role in post-transcriptional re...

  18. Preliminary results on predation of gypsy moth egg masses in Slovakia

    Treesearch

    Marek Turcani; Andrew Liebhold; Michael McManus; Julius Novotny

    2003-01-01

    Predation of gypsy moth egg masses was studied in Slovakia from 1999-2002. Predation on naturally laid egg masses was recorded and linear regression was used to test the hypothesis that predation follows a type II vs. type III functional response. We also investigated the role of egg mass predation in gypsy moth population dynamics. The relative contribution of...

  19. Identification, expression and phylogenetic analysis of EgG1Y162 from Echinococcus granulosus.

    PubMed

    Zhang, Fengbo; Ma, Xiumin; Zhu, Yuejie; Wang, Hongying; Liu, Xianfei; Zhu, Min; Ma, Haimei; Wen, Hao; Fan, Haining; Ding, Jianbing

    2014-01-01

    This study was to clone, identify and analyze the characteristics of egG1Y162 gene from Echinococcus granulosus. Genomic DNA and total RNAs were extracted from four different developmental stages of protoscolex, germinal layer, adult and egg of Echinococcus granulosus, respectively. Fluorescent quantitative PCR was used for analyzing the expression of egG1Y162 gene. Prokaryotic expression plasmid of pET41a-EgG1Y162 was constructed to express recombinant His-EgG1Y162 antigen. Western blot analysis was performed to detect antigenicity of EgG1Y162 antigen. Gene sequence, amino acid alignment and phylogenetic tree of EgG1Y162 were analyzed by BLAST, online Spidey and MEGA4 software, respectively. EgG1Y162 gene was expressed in four developmental stages of Echinococcus granulosus. And, egG1Y162 gene expression was the highest in the adult stage, with the relative value of 19.526, significantly higher than other three stages. Additionally, Western blot analysis revealed that EgG1Y162 recombinant protein had good reaction with serum samples from Echinococcus granulosus infected human and dog. Moreover, EgG1Y162 antigen was phylogenetically closest to EmY162 antigen, with the similarity over 90%. Our study identified EgG1Y162 antigen in Echinococcus granulosus for the first time. EgG1Y162 antigen had a high similarity with EmY162 antigen, with the genetic differences mainly existing in the intron region. And, EgG1Y162 recombinant protein showed good antigenicity.

  20. Dynamics of bacterial and fungal communities associated with eggshells during incubation

    PubMed Central

    Grizard, Stéphanie; Dini-Andreote, Francisco; Tieleman, B Irene; Salles, Joana F

    2014-01-01

    Microorganisms are closely associated with eggs and may play a determinant role in embryo survival. Yet, the majority of studies focusing on this association relied on culture-based methodology, eventually leading to a skewed assessment of microbial communities. By targeting the 16S rRNA gene and internal transcribed spacer (ITS) region, we, respectively, described bacterial and fungal communities on eggshells of the homing pigeon Columba livia. We explored their structure, abundance, and composition. Firstly, we showed that sampling technique affected the outcome of the results. While broadly used, the egg swabbing procedure led to a lower DNA extraction efficiency and provided different profiles of bacterial communities than those based on crushed eggshell pieces. Secondly, we observed shifts in bacterial and fungal communities during incubation. At late incubation, bacterial communities showed a reduction in diversity, while their abundance increased, possibly due to the competitive advantage of some species. When compared to their bacterial counterparts, fungal communities also decreased in diversity at late incubation. In that case, however, the decline was associated with a diminution of their overall abundance. Conclusively, our results showed that although incubation might inhibit microbial growth when compared to unincubated eggs, we observed the selective growth of specific bacterial species during incubation. Moreover, we showed that fungi are a substantial component of the microbial communities associated with eggshells and require further investigations in avian ecology. Identifying the functional roles of these microorganisms is likely to provide news insights into the evolutionary strategies that control embryo survival. We aimed to describe the dynamics of bacterial and fungal communities on homing pigeon eggshell surfaces. We investigated these communities at early and late incubation stages. PMID:24772289

  1. Development and validation of LC-MS/MS method for the determination of Ochratoxin A and its metabolite Ochratoxin α in poultry tissues and eggs.

    PubMed

    Paoloni, Angela; Solfrizzo, Michele; Bibi, Rita; Pecorelli, Ivan

    2018-05-04

    The objective of this study was to develop a liquid chromatography tandem mass spectrometry (LC-MS/MS) method for the determination of Ochratoxin A (OTA) and Ochratoxin α (OTα) in poultry tissues and eggs. The two toxins were extracted by a mixture of acetonitrile/water, purified with a reversed phase C18 solid phase extraction column (SPE) and determined by LC-MS/MS. The LC-MS/MS method performances were evaluated in terms of linearity in solvent and in matrix (ranged from 0.5 to 15.10 µg L -1 for OTA and from 0.60 to 17.85 µg L -1 for OTα), limit of detection (LOD), limit of quantitation (LOQ), specificity, accuracy and precision in repeatability conditions. Recovery experiments were performed by spiking poultry liver, kidney, muscle and eggs around 1 µg kg -1 and 10 µg kg -1 . LODs were 0.27 and 0.26 µg kg -1 while LOQs were fixed at 1.0 and 1.2 µg kg -1 for OTA and OTα, respectively. Main recoveries for OTA ranged from 82 to 109% and for OTα ranged from 55 to 89%. The values of within-laboratory relative standard deviation (RSD r ) were equal to or below 20%. Considering the results obtained and that all analytical performance criteria were fulfilled, the new extraction and purification method developed for OTA and OTα determination in animal tissues and eggs was found appropriate for control laboratories and research activities designed to ensure food safety.

  2. Differential immunosuppression by Campoletis chlorideae eggs and ichnovirus in larvae of Helicoverpa armigera and Spodoptera exigua.

    PubMed

    Han, Li-Bin; Yin, Li-Hong; Huang, Ling-Qiao; Wang, Chen-Zhu

    2015-09-01

    The ichneumonid wasp, Campoletis chlorideae Uchida, successfully develops in the cotton bollworm Helicoverpa armigera (Hübner), but rarely survives in the beet armyworm Spodoptera exigua (Hübner) due to the encapsulation by host immunity. In this study, we investigated the role of C. chlorideae ichnovirus (CcIV) and eggs in the evasion of the host immune system. Washed eggs of different types, immature, mature, newly laid, or pretreated with protease K, were injected alone or with the calyx fluid containing CcIV into the larvae of H. armigera and S. exigua. In H. armigera, when injected with washed eggs alone, only 9.5% of the mature eggs were encapsulated at 24h post-injection. This is much lower than that of the immature eggs (100%), mature eggs pretreated with protease K (100%) and newly laid eggs (54.4%). No encapsulation was observed when the washed eggs were co-injected with calyx fluid at 24h post-injection. Conversely, the eggs in all treatments were encapsulated in S. exigua. Electron microscopic observations of parasitoid eggs showed structural differences between the surfaces of the mature and other kinds of eggs. The injected CcIV decreased the numbers of host hemocytes and suppressed the spreading ability of plasmatocytes and granulocytes in H. armigera, but had little effect on the hemocytes from S. exigua. In conclusion, the C. chlorideae egg provides a passive protection against encapsulation by itself, and CcIV supplies an active protection through disrupting host immune responses. These coordinated protections are host-specific, implying their role in host range determination. Copyright © 2015 Elsevier Inc. All rights reserved.

  3. Impact of egg disinfection of hatching eggs on the eggshell microbiome and bacterial load.

    PubMed

    Olsen, R; Kudirkiene, E; Thøfner, I; Pors, S; Karlskov-Mortensen, P; Li, L; Papasolomontos, S; Angastiniotou, C; Christensen, J

    2017-09-01

    Disinfection of hatching eggs is essential to ensure high quality production of broilers. Different protocols are followed in different hatcheries; however, only limited scientific evidence on how the disinfection procedures impact the microbiome is available. The aim of the present study was to characterize the microbiome and aerobic bacterial load of hatching eggs before disinfection and during the subsequent disinfection steps. The study included a group of visibly clean and a group of visibly dirty eggs. For dirty eggs, an initial wash in chlorine was performed, hereafter all eggs were submitted to two times fumigation and finally spray disinfection. The eggshell microbiome was characterized by sequencing of the total amount of 16S rRNA extracted from each sample, consisting of shell surface swabs of five eggs from the same group. In addition, the number of colony forming units (cfu) under aerobic conditions was established for each disinfection step. The disinfection procedure reduced the bacterial load from more than 104 cfu (initially visibly clean eggs) and 105 cfu (initially visibly dirty eggs) to less than 10 cfu per sample after disinfection for both groups of eggs. The microbiome of both initially visibly clean and initially visibly dirty eggs had the highest abundances of the phyla Firmicutes, Proteobacteria and Bacteroidetes. Within the phyla Firmicutes the relative abundances of Clostridiales decreased while Lactobacillus increased from before to after final disinfection. In conclusion, the investigated disinfection procedure is effective in reducing the bacterial load, and by adding a chlorine wash for initially visibly dirty eggs, the microbiome of initially visibly clean and initially visibly dirty eggs had a highly similar microflora after the final disinfection step. © 2017 Poultry Science Association Inc.

  4. The effect of fermented buckwheat on producing l-carnitine- and γ-aminobutyric acid (GABA)-enriched designer eggs.

    PubMed

    Park, Namhyeon; Lee, Tae-Kyung; Nguyen, Thi Thanh Hanh; An, Eun-Bae; Kim, Nahyun M; You, Young-Hyun; Park, Tae-Sub; Kim, Doman

    2017-07-01

    The potential of fermented buckwheat as a feed additive was studied to increase l-carnitine and γ-aminobutyric acid (GABA) in designer eggs. Buckwheat contains high levels of lysine, methionine and glutamate, which are precursors for the synthesis of l-carnitine and GABA. Rhizopus oligosporus was used for the fermentation of buckwheat to produce l-carnitine and GABA that exert positive effects such as enhanced metabolism, antioxidant activities, immunity and blood pressure control. A novel analytical method for simultaneously detecting l-carnitine and GABA was developed using liquid chromatography/mass spectrometry (LC/MS) and LC/MS/MS. The fermented buckwheat extract contained 4 and 34 times more l-carnitine and GABA respectively compared with normal buckwheat. Compared with the control, the fermented buckwheat extract-fed group showed enriched l-carnitine (13.6%) and GABA (8.4%) in the yolk, though only l-carnitine was significantly different (P < 0.05). Egg production (9.4%), albumen weight (2.1%) and shell weight (5.8%) were significantly increased (P < 0.05). There was no significant difference in yolk weight, and total cholesterol (1.9%) and triglyceride (4.9%) in the yolk were lowered (P < 0.05). Fermented buckwheat as a feed additive has the potential to produce l-carnitine- and GABA-enriched designer eggs with enhanced nutrition and homeostasis. These designer eggs pose significant potential to be utilized in superfood production and supplement industries. © 2016 Society of Chemical Industry. © 2016 Society of Chemical Industry.

  5. Impacts of toxic thresholds of sediment-associated contaminants to robust redhorse (Moxostoma robustum) in the Lower Oconee River

    USGS Publications Warehouse

    Lasier, P.; Winger, P.; Bogenrieder, K.; Shelton, J.

    2000-01-01

    The robust redhorse is a ?Species-at-Risk? in the lower Oconee River, GA. The population is composed of aging adults with little natural recruitment. Factors contributing to the loss of early-life stages are unknown, but contaminants associated with fine sediments may play a role. The objectives of this study were to determine toxicities of sediments and pore waters from the Oconee River to early-life stages of robust redhorse and to establish toxic thresholds of metals (Cd, Cu, Mn, Zn) and ammonia, elements potentially threatening this species. Depositional sediments were collected from the only known spawning site and three sites downstream of major tributaries. Sediment pore waters were extracted in the laboratory from all sites and in situ at two sites. Toxicity tests with sediments, pore waters and metal solutions were initiated with eggs, yolk-sac fry and swim-up fry to determine effects on the life stage initially exposed as well as effects manifested in later developmental stages. Survival and growth were test endpoints, and toxicity was observed in both sediments and pore waters. Although the yolk- sac stage was the most sensitive across all tests, sediment toxicity was elicited only in tests initiated with eggs that developed through the yolk-sac stage. Toxicity appeared to be due to Mn in sediment and pore water exposures, but was more prevalent in pore waters. Sediment handling and the associated effects on redox potential contributed to the elevated concentrations of Mn in pore waters. Pore waters extracted in situ had significantly less Mn and were less toxic than laboratory-extracted pore waters. These data suggest that sediment-associated Mn may impact early-life stages of robust redhorse in the Oconee River.

  6. Protein Phosphatase 2A Antagonizes ATM and ATR in a Cdk2- and Cdc7-Independent DNA Damage Checkpoint

    PubMed Central

    Petersen, Paris; Chou, Danny M.; You, Zhongsheng; Hunter, Tony; Walter, Johannes C.; Walter, Gernot

    2006-01-01

    We previously used a soluble cell-free system derived from Xenopus eggs to investigate the role of protein phosphatase 2A (PP2A) in chromosomal DNA replication. We found that immunodepletion of PP2A or inhibition of PP2A by okadaic acid (OA) inhibits initiation of DNA replication by preventing loading of the initiation factor Cdc45 onto prereplication complexes. Evidence was provided that PP2A counteracts an inhibitory protein kinase that phosphorylates and inactivates a crucial Cdc45 loading factor. Here, we report that the inhibitory effect of OA is abolished by caffeine, an inhibitor of the checkpoint kinases ataxia-telangiectasia mutated protein (ATM) and ataxia-telangiectasia related protein (ATR) but not by depletion of ATM or ATR from the extract. Furthermore, we demonstrate that double-strand DNA breaks (DSBs) cause inhibition of Cdc45 loading and initiation of DNA replication and that caffeine, as well as immunodepletion of either ATM or ATR, abolishes this inhibition. Importantly, the DSB-induced inhibition of Cdc45 loading is prevented by addition of the catalytic subunit of PP2A to the extract. These data suggest that DSBs and OA prevent Cdc45 loading through different pathways, both of which involve PP2A, but only the DSB-induced checkpoint implicates ATM and ATR. The inhibitory effect of DSBs on Cdc45 loading does not result from downregulation of cyclin-dependent kinase 2 (Cdk2) or Cdc7 activity and is independent of Chk2. However, it is partially dependent on Chk1, which becomes phosphorylated in response to DSBs. These data suggest that PP2A counteracts ATM and ATR in a DNA damage checkpoint in Xenopus egg extracts. PMID:16479016

  7. Chum salmon egg extracts induce upregulation of collagen type I and exert antioxidative effects on human dermal fibroblast cultures.

    PubMed

    Yoshino, Atsushi; Polouliakh, Natalia; Meguro, Akira; Takeuchi, Masaki; Kawagoe, Tatsukata; Mizuki, Nobuhisa

    2016-01-01

    Components of fish roe possess antioxidant and antiaging activities, making them potentially very beneficial natural resources. Here, we investigated chum salmon eggs (CSEs) as a source of active ingredients, including vitamins, unsaturated fatty acids, and proteins. We incubated human dermal fibroblast cultures for 48 hours with high and low concentrations of CSE extracts and analyzed changes in gene expression. Cells treated with CSE extract showed concentration-dependent upregulation of collagen type I genes and of multiple antioxidative genes, including OXR1, TXNRD1, and PRDX family genes. We further conducted in silico phylogenetic footprinting analysis of promoter regions. These results suggested that transcription factors such as acute myeloid leukemia-1a and cyclic adenosine monophosphate response element-binding protein may be involved in the observed upregulation of antioxidative genes. Our results support the idea that CSEs are strong candidate sources of antioxidant materials and cosmeceutically effective ingredients.

  8. Scrambled eggs: Proteomic portraits and novel biomarkers of egg quality in zebrafish (Danio rerio)

    PubMed Central

    Yilmaz, Ozlem; Patinote, Amélie; Nguyen, Thao Vi; Com, Emmanuelle; Lavigne, Regis; Pineau, Charles; Sullivan, Craig V.; Bobe, Julien

    2017-01-01

    Egg quality is a complex biological trait and a major determinant of reproductive fitness in all animals. This study delivered the first proteomic portraits of egg quality in zebrafish, a leading biomedical model for early development. Egg batches of good and poor quality, evidenced by embryo survival for 24 h, were sampled immediately after spawning and used to create pooled or replicated sample sets whose protein extracts were subjected to different levels of fractionation before liquid chromatography and tandem mass spectrometry. Obtained spectra were searched against a zebrafish proteome database and detected proteins were annotated, categorized and quantified based on normalized spectral counts. Manually curated and automated enrichment analyses revealed poor quality eggs to be deficient of proteins involved in protein synthesis and energy and lipid metabolism, and of some vitellogenin products and lectins, and to have a surfeit of proteins involved in endo-lysosomal activities, autophagy, and apoptosis, and of some oncogene products, lectins and egg envelope proteins. Results of pathway and network analyses suggest that this aberrant proteomic profile results from failure of oocytes giving rise to poor quality eggs to properly transit through final maturation, and implicated Wnt signaling in the etiology of this defect. Quantitative comparisons of abundant proteins in good versus poor quality eggs revealed 17 candidate egg quality markers. Thus, the zebrafish egg proteome is clearly linked to embryo developmental potential, a phenomenon that begs further investigation to elucidate the root causes of poor egg quality, presently a serious and intractable problem in livestock and human reproductive medicine. PMID:29145436

  9. Scrambled eggs: Proteomic portraits and novel biomarkers of egg quality in zebrafish (Danio rerio).

    PubMed

    Yilmaz, Ozlem; Patinote, Amélie; Nguyen, Thao Vi; Com, Emmanuelle; Lavigne, Regis; Pineau, Charles; Sullivan, Craig V; Bobe, Julien

    2017-01-01

    Egg quality is a complex biological trait and a major determinant of reproductive fitness in all animals. This study delivered the first proteomic portraits of egg quality in zebrafish, a leading biomedical model for early development. Egg batches of good and poor quality, evidenced by embryo survival for 24 h, were sampled immediately after spawning and used to create pooled or replicated sample sets whose protein extracts were subjected to different levels of fractionation before liquid chromatography and tandem mass spectrometry. Obtained spectra were searched against a zebrafish proteome database and detected proteins were annotated, categorized and quantified based on normalized spectral counts. Manually curated and automated enrichment analyses revealed poor quality eggs to be deficient of proteins involved in protein synthesis and energy and lipid metabolism, and of some vitellogenin products and lectins, and to have a surfeit of proteins involved in endo-lysosomal activities, autophagy, and apoptosis, and of some oncogene products, lectins and egg envelope proteins. Results of pathway and network analyses suggest that this aberrant proteomic profile results from failure of oocytes giving rise to poor quality eggs to properly transit through final maturation, and implicated Wnt signaling in the etiology of this defect. Quantitative comparisons of abundant proteins in good versus poor quality eggs revealed 17 candidate egg quality markers. Thus, the zebrafish egg proteome is clearly linked to embryo developmental potential, a phenomenon that begs further investigation to elucidate the root causes of poor egg quality, presently a serious and intractable problem in livestock and human reproductive medicine.

  10. New multiplex PCR method for the simultaneous diagnosis of the three known species of equine tapeworm.

    PubMed

    Bohórquez, G Alejandro; Luzón, Mónica; Martín-Hernández, Raquel; Meana, Aránzazu

    2015-01-15

    Although several techniques exist for the detection of equine tapeworms in serum and feces, the differential diagnosis of tapeworm infection is usually based on postmortem findings and the morphological identification of eggs in feces. In this study, a multiplex polymerase chain reaction (PCR)-based method for the simultaneuos detection of Anoplocephala magna, Anoplocephala perfoliata and Anoplocephaloides mamillana has been developed and validated. The method simultaneously amplifies hypervariable SSUrRNA gene regions in the three tapeworm species in a single reaction using three pairs of primers, which exclusively amplify the internal transcribed spacer 2 (ITS-2) in each target gene. The method was tested on three types of sample: (a) 1/10, 1/100, 1/500, 1/1000, 1/2000 and 1/5000 dilutions of 70 ng of genomic DNA of the three tapeworm species, (b) DNA extracted from negative aliquots of sediments of negative control fecal samples spiked with 500, 200, 100, 50 and 10 eggs (only for A. magna and A. perfoliata; no A. mamillana eggs available) and (c) DNA extracted from 80, 50, 40, 30, 10 and 1 egg per 2 μl of PCR reaction mix (only for A. magna and A. perfoliata; no A. mamillana eggs available). No amplification was observed against the DNA of Gasterophilus intestinalis, Parascaris equorum and Strongylus vulgaris. The multiplex PCR method emerged as specific for the three tapeworms and was able to identify as few as 50 eggs per fecal sample and as little as 0.7 ng of control genomic DNA obtained from the three species. The method proposed is able to differentiate infections caused by the two most frequent species A. magna or A. perfoliata when the eggs are present in feces and is also able to detect mixed infections by the three cestode species. Copyright © 2014 Elsevier B.V. All rights reserved.

  11. HPLC-DAD Analysis and In-Vitro Property of Polyphenols Extracts from (Solanum Aethiopium) Fruits on α -Amylase, α -Glucosidase and Angiotensin - 1- Converting Enzyme Activities

    PubMed Central

    Nwanna, E. E; Ibukun, E. O; Oboh, G.; Ademosun, A. O.; Boligon, A. A.; Athayde, M.

    2014-01-01

    AIM: Garden egg (Solanum aethiopium) is an edible fruits vegetable with  different species.This study investigated characterisation and the effect of the phenolics extracts from S. aethiopium species with enzymes linked with type -2-diabetes (α-amylase and α-glucosidase) and hypertension [Angiotensin-1-converting enzyme (ACE)]. METHODS: Fresh samples of the 5 species of the garden egg namely, [Solanum gilo (PW), Solanum torvum (TWS), Solanum kumba (PGR), Solanum incanum (GSB), and Solanum indicum (WSB)] were oven-dried at 50°C and milled into flour. The aqueous extracts were prepared (1:50 w/v). The phenolic contents (total phenol and total flavonoid), vitamin C and 1,1-diphenyl–2-picrylhydrazyl (DPPH), the antioxidant activities of the extracts were evaluated. The ability of the extracts to inhibit diabetes enzymes in rat pancreas as well as the inhibition of angiotensin-1-converting (ACE) enzyme in lungs homogenates in vitro were investigated. Furthermore, the fruits polyphenols were identified and quantified using HPLC-DAD. RESULTS: The phenolic contents ranged from 2.70-3.76 mgGAE/g, while there were no significant (P>0.05) differences in their flavonoid content and ability to reduce Fe3+ to Fe2+. The vitamin C contents of the species ranged from 4.01-6.52 mg/ml. The extracts scavenged DPPH in a dose dependent manner with the IC50 values ranging from 3.23-4.20 mg/ml. Furthermore, the extracts showed strong inhibition of α-glucosidase, mild inhibition of α-amylase and strong inhibition of ACE activities. CONCLUSION: This study showed that the inhibition of the key enzymes relevant to type-2 diabetes and hypertension could be part of the mechanisms by which garden egg manage/prevent the degenerative conditions. PMID:25598760

  12. Nest Sanitation as the Evolutionary Background for Egg Ejection Behaviour and the Role of Motivation for Object Removal

    PubMed Central

    Poláček, Miroslav; Griggio, Matteo; Bartíková, Michaela; Hoi, Herbert

    2013-01-01

    Higher interclutch colour variation can evolve under the pressure of brood parasitism to increase the detection of parasitic eggs. Nest sanitation could be a prerequisite for the evolution of anti-parasite defence in terms of egg ejection. In this respect, we used nest sanitation behaviour as a tool to identify: i) motivation and its underlying function and, ii) which features provoke ejection behaviour. Therefore, we experimentally tested whether size, colour or shape may influence ejection behaviour using artificial flat objects. We found a high interclutch variation in egg colouration and egg size in our tree sparrow (Passer montanus) population. Using colour and size we were in fact able to predict clutch affiliation for each egg. Our experiments further revealed the existence of direct anti-parasite behaviours and birds are able to recognise conspecific eggs, since only experimentally-deposited eggs have been removed. Moreover, experiments with different objects revealed that the motivation of tree sparrows to remove experimental objects from their nests was highest during egg laying for objects of varying size, most likely because of parasitism risk at this breeding stage. In contrary, motivation to remove white objects and objects with edges was higher during incubation stage as behavioural patterns connected to hatching started to emerge. The fact that rejection rate of our flat objects was higher than real egg ejection, suggests that egg ejection in tree sparrows and probably more general in small passerines, to be limited by elevated costs to eject eggs with their beaks. The presence of anti-parasite behaviour supports our suggestion that brood parasitism causes variation in egg features, as we have found that tree sparrows can recognise and reject conspecific eggs in their clutch. In conclusion, in tree sparrows it seems that nest sanitation plays a key role in the evolution of the removal of parasitic eggs. PMID:24223165

  13. Nest sanitation as the evolutionary background for egg ejection behaviour and the role of motivation for object removal.

    PubMed

    Poláček, Miroslav; Griggio, Matteo; Bartíková, Michaela; Hoi, Herbert

    2013-01-01

    Higher interclutch colour variation can evolve under the pressure of brood parasitism to increase the detection of parasitic eggs. Nest sanitation could be a prerequisite for the evolution of anti-parasite defence in terms of egg ejection. In this respect, we used nest sanitation behaviour as a tool to identify: i) motivation and its underlying function and, ii) which features provoke ejection behaviour. Therefore, we experimentally tested whether size, colour or shape may influence ejection behaviour using artificial flat objects. We found a high interclutch variation in egg colouration and egg size in our tree sparrow (Passer montanus) population. Using colour and size we were in fact able to predict clutch affiliation for each egg. Our experiments further revealed the existence of direct anti-parasite behaviours and birds are able to recognise conspecific eggs, since only experimentally-deposited eggs have been removed. Moreover, experiments with different objects revealed that the motivation of tree sparrows to remove experimental objects from their nests was highest during egg laying for objects of varying size, most likely because of parasitism risk at this breeding stage. In contrary, motivation to remove white objects and objects with edges was higher during incubation stage as behavioural patterns connected to hatching started to emerge. The fact that rejection rate of our flat objects was higher than real egg ejection, suggests that egg ejection in tree sparrows and probably more general in small passerines, to be limited by elevated costs to eject eggs with their beaks. The presence of anti-parasite behaviour supports our suggestion that brood parasitism causes variation in egg features, as we have found that tree sparrows can recognise and reject conspecific eggs in their clutch. In conclusion, in tree sparrows it seems that nest sanitation plays a key role in the evolution of the removal of parasitic eggs.

  14. Effects of Toxic Compounds in Montipora capitata on Exogenous and Endogenous Zooxanthellae Performance and Fertilization Success

    PubMed Central

    Hagedorn, Mary; Farrell, Ann; Carter, Virginia; Zuchowicz, Nikolas; Johnston, Erika; Padilla-Gamiño, Jacqueline; Gunasekera, Sarath; Paul, Valerie

    2015-01-01

    Studies have identified chemicals within the stony coral genus Montipora that have significant biological activities. For example, Montiporic acids A and B and other compounds have been isolated from the adult tissue and eggs of Montipora spp. and have displayed antimicrobial activity and cytotoxicity in cultured cells. The ecological role of these toxic compounds is currently unclear. This study examines the role these toxins play in reproduction. Toxins were found in the eggs and larvae of the coral Montipora capitata. Releasing these toxins by crushing both the eggs and larvae resulted in irreversible inhibition of photosynthesis in endogenous and exogenous zooxanthellae within minutes. Moreover, these toxins were stable, as frozen storage of eggs and larvae did not affect toxicity. Photosynthetic competency of Porites compressa zooxanthellae treated with either frozen or fresh, crushed eggs was inhibited similarly (P > 0.05, ANCOVA). Addition of toxic eggs plugs to live P. compressa fragments caused complete tissue necrosis under the exposed area on the fragments within 1 week. Small volumes of M. capitata crushed eggs added to sperm suspensions reduced in vitro fertilization success by killing the sperm. After 30 min, untreated sperm maintained 90 ± 1.9% SEM motility while those treated with crushed eggs were rendered immotile, 4 ± 1.4% SEM. Flow cytometry indicated membrane disruption of the immotile sperm. Fertilization success using untreated sperm was 79 ± 4% SEM, whereas the success rate dropped significantly after exposure to the crushed eggs, 1.3 ± 0% SEM. Unlike the eggs and the larvae, M. capitata sperm did not reduce the photosynthetic competency of P. compressa zooxanthellae, suggesting the sperm was nontoxic. The identity of the toxins, cellular mechanism of action, advantage of the toxins for M. capitata and their role on the reef are still unknown. PMID:25714606

  15. A simple and efficient method for preparing partially purified phosvitin from egg yolk using ethanol and salts.

    PubMed

    Ko, K Y; Nam, K C; Jo, C; Lee, E J; Ahn, D U

    2011-05-01

    The objective of this study was to develop a new protocol that could be used for large-scale separation of phosvitin from egg yolk using ethanol and salts. Yolk granules, which contain phosvitin, were precipitated after diluting egg yolk with 9 volumes of distilled water. The pH of the yolk solution was adjusted to pH 4.0 to 8.0 using 6 N HCl or NaOH, and then yolk granules containing phosvitin was separated by centrifugation at 3,220 × g for 30 min. Lipids and phospholipids were removed from the insoluble yolk granules using 85% ethanol. The optimal volumes and concentration of ethanol in removing lipids from the precipitants were determined. After centrifugation, the lipid-free precipitants were homogenized with 9 volumes of ammonium sulfate [(NH(4))(2)SO(4)] or NaCl to extract phosvitin. The optimal pH and concentration of (NH(4))(2)SO(4) or NaCl for the highest recovery rate and purity for phosvitin in final solution were determined. At pH 6.0, all the phosvitin in diluted egg yolk solution was precipitated. Among the (NH(4))(2)SO(4) and NaCl conditions tested, 10% (NH(4))(2)SO(4) or 10% NaCl at pH 4.0 yielded the greatest phosvitin extraction from the lipid-free precipitants. The recovery rates of phosvitin using (NH(4))(2)SO(4) and NaCl were 72 and 97%, respectively, and their purity was approximately 85%. Salt was removed from the extract using ultrafiltration. The salt-free phosvitin solution was concentrated using ultrafiltration, the impurities were removed by centrifugation, and the resulting solution was freeze-dried. The partially purified phosvitin was suitable for human use because ethanol was the only solvent used to remove lipids, (NH(4))(2)SO(4) or NaCl was used to extract phosvitin, and ultrafiltration was used to remove salt and concentrate the extract. The developed method was simple and suitable for a large-scale preparation of partially purified phosvitin.

  16. Occurrence of sweet refuse at disposal sites: rainwater retention capacity and potential breeding opportunities for Aedes aegypti.

    PubMed

    Dieng, Hamady; Satho, Tomomitsu; Meli, Nur Khairatun Khadijah Binti; Abang, Fatimah; Nolasco-Hipolito, Cirilo; Hakim, Hafijah; Miake, Fumio; Zuharah, Wan Fatma; Kassim, Nur Faeza A; Ab Majid, Abdul Hafiz; Morales Vargas, Ronald E; Morales, Noppawan P; Noweg, Gabriel Tonga

    2018-05-01

    Nectar is the staple diet of adult mosquitoes in the wild, but its availability is inconsistent and can be affected by rainfall. In urban centers, Aedes vectors commonly use man-made containers as their major habitat; however, they can colonize any items replenished by rainfall. Garbage output has increased significantly in recent years, at a time when collection frequency is reducing. Such garbage usually includes organic components, some of which are sweet and can be fed upon by other animals or become can containers for rainwater. Despite evidence that Aedes larvae can thrive in containers comprised of organic waste material, which can be produced by rodents gnawing on fruits or vegetables, and that adults can survive on sweet waste fluids, the capacity of organic waste materials to accumulate rainwater and act as egg deposition sites has not been examined. It is also unknown for how long sweet extracts can sustain the life of adult vectors. Here, we investigated the abundance of sweet leftovers at garbage sites and the rainwater retention capacity of some organic materials through a field survey and laboratory bioassays. We also examined whether sweet waste fluids impact egg hatching success and longevity of Aedes aegypti. The results of this study indicated that sweet products with leftovers are highly prevalent in garbage. When exposed to rain, food items (BAFrc, banana fruit resembling container; and BSPrc, boiled sweet potato resembling container) and the packaging of sweet foods (SMIc, sweetened condensed milk can) retained water. When provided an opportunity to oviposit in cups containing BAF extract (BAFex), BSP extract (BSPex), and SMI extract (SMIex), eggs were deposited in all media. Egg maturation in the BAFex environment resulted in similar larval eclosion success to that resulting from embryo development in a water milieu. Adults maintained on sweet waste extracts had long lifespans, although shorter than that of their sugar solution (SUS)-fed counterparts. Taken together, these results indicated that sweet waste materials are useful to dengue mosquitoes, acting both as oviposition sites and energy sources.

  17. DNA of Dientamoeba fragilis detected within surface-sterilized eggs of Enterobius vermicularis.

    PubMed

    Röser, Dennis; Nejsum, Peter; Carlsgart, Anne Josefine; Nielsen, Henrik Vedel; Stensvold, Christen Rune

    2013-01-01

    With no evidence of a cyst stage, the mode of transmission of Dientamoeba fragilis, an intestinal protozoon of common occurrence and suggested pathogenicity, is incompletely known. Numerous studies have suggested that eggs of intestinal nematodes, primarily Enterobius vermicularis (pinworm), can serve as vectors for D. fragilis, although attempts to culture D. fragilis from pinworm eggs have been unsuccessful and data from epidemiological studies on D. fragilis/pinworm co-infection have been conflicting. The aim of this study was to investigate whether we could detect D. fragilis DNA from pinworm eggs collected from routine diagnostic samples (cellophane tape) and surface-sterilised by hypochlorite. DNA was extracted from individual eggs and tested by PCR using D. fragilis- and E. vermicularis-specific primers; amplicons were sequenced for confirmation. In cellophane tape samples from 64 patients with unknown D. fragilis status we detected D. fragilis DNA in 12/238 (5%) eggs, and in a patient known to harbour D. fragilis we detected D. fragilis DNA in 39/99 (39%) eggs. The finding of D. fragilis DNA within eggs of E. vermicularis strongly supports the hypothesis of D. fragilis-transmission by pinworm and has implications for antimicrobial intervention as well as control and public health measures. Copyright © 2012 Elsevier Inc. All rights reserved.

  18. Worker reproduction of the invasive yellow crazy ant Anoplolepis gracilipes.

    PubMed

    Lee, Ching-Chen; Nakao, Hirotaka; Tseng, Shu-Ping; Hsu, Hung-Wei; Lin, Gwo-Li; Tay, Jia-Wei; Billen, Johan; Ito, Fuminori; Lee, Chow-Yang; Lin, Chung-Chi; Yang, Chin-Cheng Scotty

    2017-01-01

    Reproductive division of labor is one of the key features of social insects. Queens are adapted for reproduction while workers are adapted for foraging and colony maintenance. In many species, however, workers retain functional ovaries and can lay unfertilized male eggs or trophic eggs. Here we report for the first time on the occurrence of physogastric workers and apparent worker reproduction in the invasive yellow crazy ant Anoplolepis gracilipes (Fr. Smith). We further examined the reproductive potential and nutritional role of physogastric workers through multidisciplinary approaches including morphological characterization, laboratory manipulation, genetic analysis and behavioral observation. Egg production with two types of eggs, namely reproductive and trophic eggs, by physogastric workers was found. The reproductive egg was confirmed to be haploid and male-destined, suggesting that the workers produced males via arrhenotokous parthenogenesis as no spermatheca was discovered. Detailed observations suggested that larvae were mainly fed with trophic eggs. Along with consumption of trophic eggs by queens and other castes as part of their diet, the vital role of physogastric workers as "trophic specialist" is confirmed. We propose that adaptive advantages derived from worker reproduction for A. gracilipes may include 1) trophic eggs provisioned by physogastric workers likely assist colonies of A. gracilipes in overcoming unfavorable conditions such as paucity of food during critical founding stage; 2) worker-produced males are fertile and thus might offer an inclusive fitness advantage for the doomed orphaned colony.

  19. The impact of N-glycosylation on conformation and stability of immunoglobulin Y from egg yolk.

    PubMed

    Sheng, Long; He, Zhenjiao; Chen, Jiahui; Liu, Yaofa; Ma, Meihu; Cai, Zhaoxia

    2017-03-01

    Immunoglobulin Y (IgY) is a new therapeutic antibody, and its applications in industry are very broad. To provide insight into the effects of N-glycosylation on IgY, its conformation and stability were studied. In this research, IgY was extracted from egg yolk and then digested by peptide-N4-(N-acetyl-beta-glucosaminyl) asparagine-amidase. SDS-PAGE and infrared absorption spectrum showed that carbohydrates were distinctly reduced after enzymolysis. The circular dichroism spectrum indicated that the IgY molecule became more flexible and disordered after removal of N-glycan. The fluorescence intensity revealed that Trp residues were buried in a more hydrophobic environment after disposal of N-glycan. Storage stability decreased with the removal of oligosaccharide chains based on size-exclusion chromatography analysis. Deglycosylated IgY exhibited less resistance to guanidine hydrochloride-induced unfolding. After deglycosylation, IgY was more sensitive to pepsin. Therefore, N-glycosylation played an important role in the maintenance of the structure and stability of IgY. Copyright © 2016 Elsevier B.V. All rights reserved.

  20. Physical basis of large microtubule aster growth

    PubMed Central

    Ishihara, Keisuke; Korolev, Kirill S; Mitchison, Timothy J

    2016-01-01

    Microtubule asters - radial arrays of microtubules organized by centrosomes - play a fundamental role in the spatial coordination of animal cells. The standard model of aster growth assumes a fixed number of microtubules originating from the centrosomes. However, aster morphology in this model does not scale with cell size, and we recently found evidence for non-centrosomal microtubule nucleation. Here, we combine autocatalytic nucleation and polymerization dynamics to develop a biophysical model of aster growth. Our model predicts that asters expand as traveling waves and recapitulates all major aspects of aster growth. With increasing nucleation rate, the model predicts an explosive transition from stationary to growing asters with a discontinuous jump of the aster velocity to a nonzero value. Experiments in frog egg extract confirm the main theoretical predictions. Our results suggest that asters observed in large fish and amphibian eggs are a meshwork of short, unstable microtubules maintained by autocatalytic nucleation and provide a paradigm for the assembly of robust and evolvable polymer networks. DOI: http://dx.doi.org/10.7554/eLife.19145.001 PMID:27892852

  1. TaqMan DNA technology confirms likely overestimation of cod (Gadus morhua L.) egg abundance in the Irish Sea: implications for the assessment of the cod stock and mapping of spawning areas using egg-based methods.

    PubMed

    Fox, C J; Taylor, M I; Pereyra, R; Villasana, M I; Rico, C

    2005-03-01

    Recent substantial declines in northeastern Atlantic cod stocks necessitate improved biological knowledge and the development of techniques to complement standard stock assessment methods (which largely depend on accurate commercial catch data). In 2003, an ichthyoplankton survey was undertaken in the Irish Sea and subsamples of 'cod-like' eggs were analysed using a TaqMan multiplex, PCR (polymerase chain reaction) assay (with specific probes for cod, haddock and whiting). The TaqMan method was readily applied to the large number of samples (n = 2770) generated during the survey and when combined with a manual DNA extraction protocol had a low failure rate of 6%. Of the early stage 'cod-like' eggs (1.2-1.75 mm diameter) positively identified: 34% were cod, 8% haddock and 58% whiting. As previous stock estimates based on egg surveys for Irish Sea cod assumed that the majority of 'cod-like' eggs were from cod, the TaqMan results confirm that there was probably substantial contamination by eggs of whiting and haddock that would have inflated estimates of the stock biomass.

  2. Identification, expression and phylogenetic analysis of EgG1Y162 from Echinococcus granulosus

    PubMed Central

    Zhang, Fengbo; Ma, Xiumin; Zhu, Yuejie; Wang, Hongying; Liu, Xianfei; Zhu, Min; Ma, Haimei; Wen, Hao; Fan, Haining; Ding, Jianbing

    2014-01-01

    Objective: This study was to clone, identify and analyze the characteristics of egG1Y162 gene from Echinococcus granulosus. Methods: Genomic DNA and total RNAs were extracted from four different developmental stages of protoscolex, germinal layer, adult and egg of Echinococcus granulosus, respectively. Fluorescent quantitative PCR was used for analyzing the expression of egG1Y162 gene. Prokaryotic expression plasmid of pET41a-EgG1Y162 was constructed to express recombinant His-EgG1Y162 antigen. Western blot analysis was performed to detect antigenicity of EgG1Y162 antigen. Gene sequence, amino acid alignment and phylogenetic tree of EgG1Y162 were analyzed by BLAST, online Spidey and MEGA4 software, respectively. Results: EgG1Y162 gene was expressed in four developmental stages of Echinococcus granulosus. And, egG1Y162 gene expression was the highest in the adult stage, with the relative value of 19.526, significantly higher than other three stages. Additionally, Western blot analysis revealed that EgG1Y162 recombinant protein had good reaction with serum samples from Echinococcus granulosus infected human and dog. Moreover, EgG1Y162 antigen was phylogenetically closest to EmY162 antigen, with the similarity over 90%. Conclusion: Our study identified EgG1Y162 antigen in Echinococcus granulosus for the first time. EgG1Y162 antigen had a high similarity with EmY162 antigen, with the genetic differences mainly existing in the intron region. And, EgG1Y162 recombinant protein showed good antigenicity. PMID:25337206

  3. Calcium and Egg Activation in Drosophila

    PubMed Central

    Sartain, Caroline V.; Wolfner, Mariana F.

    2012-01-01

    Summary In many animals, a rise in intracellular calcium levels is the trigger for egg activation, the process by which an arrested mature oocyte transitions to prepare for embryogenesis. In nearly all animals studied to date, this calcium rise, and thus egg activation, is triggered by the fertilizing sperm. However in the insects that have been examined, fertilization is not necessary to activate their oocytes. Rather, these insects’ eggs activate as they transit through the female’s reproductive tract, regardless of male contribution. Recent studies in Drosophila have shown that egg activation nevertheless requires calcium and that the downstream events and molecules of egg activation are also conserved, despite the difference in initial trigger. Genetic studies have uncovered essential roles for the calcium-dependent enzyme calcineurin and its regulator calcipressin, and have hinted at roles for calmodulin, in Drosophila egg activation. Physiological and in vitro studies have led to a model in which mechanical forces that impact the Drosophila oocyte as it moves through the reproductive tract triggers the influx of calcium from the external environment, thereby initiating egg activation. Future research will aim to test this model, as well as to determine the spatiotemporal dynamics of cytoplasmic calcium flux and mode of signal propagation in this unique system. PMID:23218670

  4. Association of Beta2-Positive Clostridium perfringens Type A With Focal Duodenal Necrosis in Egg-Laying Chickens in the United States.

    PubMed

    França, M; Barrios, M A; Stabler, L; Zavala, Guillermo; Shivaprasad, H L; Lee, M D; Villegas, A M; Uzal, Francisco A

    2016-03-01

    Focal duodenal necrosis (FDN) is a poorly understood intestinal disease of egg layers, and has been associated with drops in egg production and decreased egg weights. The etiology of this disease is still unknown, but the condition has been associated with Clostridium colinum and Clostridium perfringens. In order to investigate the etiology, duodenal samples were taken from hens with FDN. The hens originated from table egg layer farms in three states. The samples were examined by histopathology, bacteriology, and immunohistochemistry. Macroscopically, all samples contained focal to multifocal, variably sized, reddened or brownish gray areas of mucosal erosion. Histopathology revealed mild to severe heterophilic and lymphoplasmacytic enteritis with loss of enterocytes at the villous tips, luminal fibrinonecrotic exudate, and variable numbers of Gram-positive and Gram-negative rod-shaped bacteria within the lesions in 16/30 samples. Clostridium perfringens was isolated by anaerobic bacteriology from 4/13 samples that had characteristic microscopic lesions of FDN. Polymerase chain reaction (PCR) revealed that all four isolates were Type A C. perfringens, positive for beta2 gene and negative for necrotic enteritis toxin B and enterotoxin genes. PCR for Clostridium colinum applied to DNA extracted from frozen intestinal samples yielded negative results in 14/14 duodenal samples. Immunohistochemistry (IHC) for 7C. perfringens, alpha and beta2 toxins stained a few to numerous long rod-shaped bacteria present in the lesions. IHC for alpha and beta2 toxins also stained enterocytes at the villous tips, inflammatory cells in the lamina propria, as well as degenerated and sloughed enterocytes present within the luminal exudate. These findings suggest that C. perfringens may play a role in the development of FDN. Experimental challenge studies with these isolates still need to be performed in order to reproduce the disease and fulfill Koch's postulates.

  5. 77 FR 70991 - Submission for OMB Review; Comment Request

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-11-28

    ... Monitoring System (NAHMS) will initiate the second national data collection of table egg layers through... have a role in the prevention and control of Samonella Enteritidis on table eggs farms. This working... a current estimate of the prevalence of Samonella Enteritidis on table egg farms. Need and Use of...

  6. Distinct roles for multiple Src family kinases at fertilization.

    PubMed

    O'Neill, Forest J; Gillett, Jessica; Foltz, Kathy R

    2004-12-01

    Egg activation at fertilization requires the release of Ca2+ from the endoplasmic reticulum of the egg. Recent evidence indicates that Src family kinases (SFKs) function in the signaling pathway that initiates this Ca2+ release in the eggs of many deuterostomes. We have identified three SFKs expressed in starfish (Asterina miniata) eggs, designated AmSFK1, AmSFK2 and AmSFK3. Antibodies made against the unique domains of each AmSFK protein revealed that all three are expressed in eggs and localized primarily to the membrane fraction. Both AmSFK1 and AmSFK3 (but not AmSFK2) are necessary for egg activation, as determined by injection of starfish oocytes with dominant-interfering Src homology 2 (SH2) domains, which specifically delay and reduce the initial release of Ca2+ at fertilization. AmSFK3 exhibits a very rapid and transient kinase activity in response to fertilization, peaking at 30 seconds post sperm addition. AmSFK1 kinase activity also increases transiently at fertilization, but peaks later, at 2 minutes. These results indicate that there are multiple SFKs present in starfish eggs with distinct, perhaps sequential, signaling roles.

  7. Structural proteins in the egg-shell of the oriental garden cricket, Gryllus mitratus

    PubMed Central

    Kawasaki, Hiroya; Sato, Hitoshi; Suzuki, Motoko

    1971-01-01

    1. The egg-shell of the oriental garden cricket, Gryllus mitratus, contained at least two different types of structural protein in an approximate ratio of 5:1. The major fraction was extracted in a solvent containing dithiothreitol, EDTA and 8m-urea, and was purified to apparent homogeneity as judged by free-boundary electrophoresis and ultracentrifugation. This was designated SH-fraction and its S-carboxymethyl derivative (CM-fraction) was also prepared. The minor fraction, insoluble in the solvent, was designated insoluble residue. 2. The major fraction was a phosphoprotein, rich in serine (29.8mol% of the total amino acids) and phosphate (nearly equimolar to serine), and O-phosphoserine was identified in its partial acid hydrolysate. The content of cystine was rather low (0.9mol%) in spite of the importance of this amino acid residue in the native form of the protein. The insoluble residue contained only a small amount of phosphorus, and its amino acid composition was clearly different from the major fraction. 3. CM-fraction, a fibrous protein with an average molecular weight of 57500, behaved as a typical polyanion owing to the high content of phosphate. SH-fraction and CM-fraction were precipitable from their aqueous solutions by the addition of bivalent metal cations, and the precipitation of CM-fraction by Ca2+ and Mg2+ was studied in detail. 4. When SH-fraction was exposed to air, intermolecular disulphide linkages were formed, yielding a net-like gel that changed its volume with changes in Ca2+, Mg2+ and Na+. 5. The possible role of this protein fraction in maintaining the integrity of the egg-shell, and a comparison of its composition and properties with other egg-shell proteins and other phosphoproteins, are discussed. ImagesFig. 2.PLATE 1 PMID:5004198

  8. The Role of Wheat and Egg Constituents in the Formation of a Covalent and Non-covalent Protein Network in Fresh and Cooked Egg Noodles.

    PubMed

    Lambrecht, Marlies A; Rombouts, Ine; Nivelle, Mieke A; Delcour, Jan A

    2017-01-01

    Noodles of constant protein content and flour-to-egg protein ratio were made with whole egg, egg white, or egg yolk. The optimal cooking time, water absorption, and cooking loss of salted whole egg noodles was respectively lower and higher than of egg white and egg yolk noodles. However, cooked whole egg noodles showed the best Kieffer-rig extensibility. Differences in noodle properties were linked to protein network formation. Disulfide bonds in whole egg noodles developed faster and to a larger extent during cooking than in egg yolk noodles but slower and to a lower extent than in egg white noodles. The balance between the rate of protein cross-linking and starch swelling determines cooked noodle properties. Ionic and hydrophobic protein interactions increase the optimum cooking time and total work in Kieffer-rig extensibility testing of fresh noodles. Hydrogen bonds and covalent cross-links are probably the main determinants of the extensibility of cooked noodles. © 2016 Institute of Food Technologists®.

  9. A male sexually dimorphic trait provides antimicrobials to eggs in blenny fish.

    PubMed

    Giacomello, Eva; Marchini, Daniela; Rasotto, Maria B

    2006-09-22

    Predation and microbial infections are the major causes of natural mortality for early life stages of oviparous species. The parental traits reducing the effects of predation are rather well described, whereas antimicrobial mechanisms enhancing offspring survival are largely unexplored. In this paper, we report that a male sexually dimorphic trait, the anal glands, of the redlip blenny (Ophioblennius atlanticus atlanticus) and the peacock blenny (Salaria pavo), two fish species with paternal egg care, produce a mucus enriched with antimicrobial substances. Histological and histochemical analyses showed that the anal glands of these species are characterized by the massive presence of mucus-secreting cells. Anal gland extracts, from both the hydrophilic and the hydrophobic protein fraction, exhibited a lysozyme-like activity. Field observations demonstrated that redlip blenny males, while performing egg care, rub the anal region over the nest internal surface, probably facilitating the transfer of mucus to eggs. These results strongly indicate that this sexually dimorphic trait is involved in egg defence against microbial infections.

  10. Alternative complement activity in the egg cytosol of amphioxus Branchiostoma belcheri: evidence for the defense role of maternal complement components.

    PubMed

    Liang, Yujun; Zhang, Shicui; Wang, Zhiping

    2009-01-01

    The eggs in most invertebrates are fertilized externally, and therefore their resulting embryos are exposed to an environment full of microbes, many of which are pathogens capable of killing other organisms. How the developing embryos of invertebrates defend themselves against pathogenic attacks is an intriguing question to biologists, and remains largely unknown. Here we clearly demonstrated that the egg cytosol prepared from the newly fertilized eggs of amphioxus Branchiostoma belcheri, an invertebrate chordate, was able to inhibit the growth of both the Gram-negative bacterium Vibrio anguillarum and the Gram-positive bacterium Staphylococcus aureus. All findings point to that it is the complement system operating via the alternative pathway that is attributable to the bacteriostatic activity. This appears to be the first report providing the evidence for the functional role of the maternal complement components in the eggs of invertebrate species, paving the way for the study of maternal immunity in other invertebrate organisms whose eggs are fertilized in vitro. It also supports the notion that the early developing embryos share some defense mechanisms common with the adult species.

  11. Avocado Seeds: Extraction Optimization and Possible Use as Antioxidant in Food

    PubMed Central

    Gómez, Francisco Segovia; Sánchez, Sara Peiró; Gallego Iradi, Maria Gabriela; Mohd Azman, Nurul Aini; Almajano, María Pilar

    2014-01-01

    Consumption of avocado (Persea americana Mill) has increased worldwide in recent years. Part of this food (skin and seed) is lost during processing. However, a high proportion of bioactive substances, such as polyphenols, remain in this residue. The primary objective of this study was to model the extraction of polyphenols from the avocado pits. In addition, a further objective was to use the extract obtained to evaluate the protective power against oxidation in food systems, as for instance oil in water emulsions and meat products. Moreover, the possible synergy between the extracts and egg albumin in the emulsions is discussed. In Response Surface Method (RSM), the variables used are: temperature, time and ethanol concentration. The results are the total polyphenols content (TPC) and the antiradical power measured by Oxygen Radical Antioxidant Capacity (ORAC). In emulsions, the primary oxidation, by Peroxide Value and in fat meat the secondary oxidation, by TBARS (Thiobarbituric acid reactive substances), were analyzed. The RSM model has an R2 of 94.69 for TPC and 96.7 for ORAC. In emulsions, the inhibition of the oxidation is about 30% for pure extracts and 60% for the combination of extracts with egg albumin. In the meat burger oxidation, the formation of TBARS is avoided by 90%. PMID:26784880

  12. Rapidly evolving zona pellucida domain proteins are a major component of the vitelline envelope of abalone eggs

    PubMed Central

    Aagaard, Jan E.; Yi, Xianhua; MacCoss, Michael J.; Swanson, Willie J.

    2006-01-01

    Proteins harboring a zona pellucida (ZP) domain are prominent components of vertebrate egg coats. Although less well characterized, the egg coat of the non-vertebrate marine gastropod abalone (Haliotis spp.) is also known to contain a ZP domain protein, raising the possibility of a common molecular basis of metazoan egg coat structures. Egg coat proteins from vertebrate as well as non-vertebrate taxa have been shown to evolve under positive selection. Studied most extensively in the abalone system, coevolution between adaptively diverging egg coat and sperm proteins may contribute to the rapid development of reproductive isolation. Thus, identifying the pattern of evolution among egg coat proteins is important in understanding the role these genes may play in the speciation process. The purpose of the present study is to characterize the constituent proteins of the egg coat [vitelline envelope (VE)] of abalone eggs and to provide preliminary evidence regarding how selection has acted on VE proteins during abalone evolution. A proteomic approach is used to match tandem mass spectra of peptides from purified VE proteins with abalone ovary EST sequences, identifying 9 of 10 ZP domain proteins as components of the VE. Maximum likelihood models of codon evolution suggest positive selection has acted among a subset of amino acids for 6 of these genes. This work provides further evidence of the prominence of ZP proteins as constituents of the egg coat, as well as the prominent role of positive selection in diversification of these reproductive proteins. PMID:17085584

  13. Ovomucoid (Gal d 1) specific IgE detected by microarray system predict tolerability to boiled hen's egg and an increased risk to progress to multiple environmental allergen sensitisation.

    PubMed

    Alessandri, C; Zennaro, D; Scala, E; Ferrara, R; Bernardi, M Livia; Santoro, M; Palazzo, P; Mari, A

    2012-03-01

    Egg allergy is a very common finding in early childhood. Detecting hen's egg (HE) allergy outgrowing and reintroduction of food containing egg is a task for the allergist. We sought to evaluate the suitability of boiled egg food challenge compared with IgE to allergenic molecules from HE white using a microarray system. Sixty-eight children referring to our centre by the family paediatricians for a suspected egg allergy were enrolled. Patients underwent double-blind, placebo-controlled food challenge with boiled and raw eggs. Challenge outcomes were compared with skin tests performed using egg white and yolk commercial extracts, to prick-prick test with boiled and raw egg white and yolk, total IgE, egg white specific IgE detected using ImmunoCAP and IgE to egg allergens available on the immunosolid phase allergen chip (ISAC) 103 microarray. Nineteen subjects (28%) were reactive to both raw and boiled egg, 14 (20.5%) to raw egg only and 35 (51.4%) tolerated both boiled and raw egg. Efficiency analysis was carried out using both raw and boiled egg challenges as gold standard. Forty four of 47 Gal d 1 negative patients tolerated boiled egg (94%). Conversely, 20 of 21 Gal d 1 positive patients reacted to raw egg (95%). None of the other tests was able to discriminate patients' response to HE challenge. Furthermore, Gal d 1 positivity seems to lead to broader environmental allergen IgE sensitization. The Gal d 1 IgE reactivity appears to be a very good predictor of HE clinical allergy. Gal d 1 positive children have a high frequency of HE allergy, whereas Gal d 1 negative children have a high frequency of tolerance to boiled egg. Multiple specific IgE detection by means of ISAC improves the diagnostic approach in HE allergic children, disclosing other food and inhalant allergic sensitizations, anyhow requiring a comprehensive clinical evaluation. © 2011 Blackwell Publishing Ltd.

  14. Correlation of polyphenolic content with radical-scavenging capacity and anthelmintic effects of Rubus ulmifolius (Rosaceae) against Haemonchus contortus.

    PubMed

    Akkari, Hafidh; Hajaji, Soumaya; B'chir, Fatma; Rekik, Mourad; Gharbi, Mohamed

    2016-05-15

    Phenolic content, antioxidant and anthelmintic activities of herbal extracts are of particular interest to drug industry; plant extracts with significant anthelmintic activity have the potential to be used as alternatives to conventional chemical drugs. In the present study, Rubus ulmifolius fruit extracts obtained using solvents of increasing polarity (water, methanol, chloroform and hexane) were examined for their antioxidant and anthelmintic activities in correlation with their polyphenolic content. In vitro antioxidant activity of all extracts was carried out using free radical-scavenging activity by 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azinobis-(3-ethilenebenzotiazolin)-6-sulfonic acid (ABTS) radical cation. In vitro anthelmintic activities were investigated on the egg and adult worms of Haemonchus contortus from sheep in comparison to albendazole. Total polyphenol content of R. ulmifolius was higher in more polar extract, ranging from 64.5 in aqueous extract to 1.57 mg gallic acid equivalents per gram of dry weight (GAE/g DW) in hexanic extract. Likewise, highest amounts of flavonoids and condensed tannins were found in aqueous extract (28.06 mg QE/g and 7.42 mg CE/g DW, respectively) compared to hexanic extract (0.71 mg QE/g and 0.29 mg CE/g DW, respectively) (p<0.05). Both DPPH and ABTS antioxidant assays showed that all tested extracts possess free radical scavenging activity, while the inhibitory concentration 50% (IC50) range values were similar for both assays (2.13-45.54 μg/mL and 1.2-43.82 μg/mL, respectively). All plant extracts showed ovicidal activity at all tested concentrations. Fruit methanolic (IC50=2.76mg/mL) and aqueous (IC50=2.08 mg/mL) extracts showed higher inhibitory effects than chloroformic (IC50=7.62 mg/mL) and hexanic (IC50=12.93 mg/mL) extracts on egg hatching (p<0.05). There was a significant correlation of total polyphenol, flavonoids and tannins content with scavenging of either DPPH (r=0.722, 0.764 and 0.752, p<0.01, respectively) or ABTS radicals (r=0.893, 0.765 and 0.722, p<0.01, respectively) and with inhibition of egg hatching (r=0.874, 0.883 and 0.862, p<0.01, respectively). Highest inhibition of motility (100%) of worms was observed 8h post-exposure in aqueous and methanolic extract at 8 mg/mL. To our knowledge, these results depict for the first time that R. ulmifolius possesses in vitro anthelmintic properties. Published by Elsevier B.V.

  15. Evolutionary stability of egg trading and parceling in simultaneous hermaphrodites: the chalk bass revisited.

    PubMed

    Crowley, Philip H; Hart, Mary K

    2007-06-07

    Several species of simultaneously hermaphroditic seabasses living on coral reefs mate by alternating male and female roles with a partner. This is known as egg trading, one of the classic and most widely cited examples of social reciprocity among animals. Some of the egg-trading seabass species, including the chalk bass, Serranus tortugarum, switch mating roles repeatedly, having subdivided their clutch of eggs into parcels offered to the partner for fertilization. Here we attempt to understand these dynamics as a pair of evolutionary games, modifying some previous approaches to better reflect the biological system. We find that the trading of egg clutches is evolutionarily stable via byproduct mutualism and resistant to invasion by rare individuals that take the male role exclusively. We note why and how parceling may reflect sexual conflict between individuals in the mating pair. We estimate evolutionarily stable parcel numbers and show how they depend on parameter values. Typically, two or more sequential parcel numbers are evolutionarily stable, though the lowest of these yields the highest fitness. Assuming that parcel numbers are adjusted to local conditions, we predict that parcel numbers in nature are inversely related both to mating group density (except at low density) and predation risk.

  16. Blue petrels recognize the odor of their egg.

    PubMed

    Leclaire, Sarah; Bourret, Vincent; Bonadonna, Francesco

    2017-09-01

    Most studies on avian olfactory communication have focused on mate choice, and the importance of olfaction in subsequent nesting stages has been poorly explored. In particular, the role of olfactory cues in egg recognition has received little attention, despite eggs potentially being spread with parental odorous secretions known to elicit individual discrimination. Here, we used behavioral choice tests to determine whether female blue petrels ( Halobaena caerulea ) can discriminate the odor of their own egg from the odor of a conspecific egg. Females preferentially approached the odor of their own egg, suggesting that blue petrels can recognize their own egg using odor cues. This finding raises the question of the adaptive value of this mechanism, and may inspire further research on odor-based egg discrimination in species suffering brood parasitism. © 2017. Published by The Company of Biologists Ltd.

  17. Cracking the egg: An insight into egg hypersensitivity.

    PubMed

    Dhanapala, Pathum; De Silva, Chamika; Doran, Tim; Suphioglu, Cenk

    2015-08-01

    Hypersensitivity to the chicken egg is a widespread disorder mainly affecting 1-2% of children worldwide. It is the second most common food allergy in children, next to cow's milk allergy. Egg allergy is mainly caused by hypersensitivity to four allergens found in the egg white; ovomucoid, ovalbumin, ovotransferrin and lysozyme. However, some research suggests the involvement of allergens exclusively found in the egg yolk such as chicken serum albumin and YGP42, which may play a crucial role in the overall reaction. In egg allergic individuals, these allergens cause conditions such as itching, atopic dermatitis, bronchial asthma, vomiting, rhinitis, conjunctivitis, laryngeal oedema and chronic urticaria, and anaphylaxis. Currently there is no permanent cure for egg allergy. Upon positive diagnosis for egg allergy, strict dietary avoidance of eggs and products containing traces of eggs is the most effective way of avoiding future hypersensitivity reactions. However, it is difficult to fully avoid eggs since they are found in a range of processed food products. An understanding of the mechanisms of allergic reactions, egg allergens and their prevalence, egg allergy diagnosis and current treatment strategies are important for future studies. This review addresses these topics and discusses both egg white and egg yolk allergy as a whole. Copyright © 2015 Elsevier Ltd. All rights reserved.

  18. Antinociceptive, anti-inflammatory and antidiabetic effects of Bryophyllum pinnatum (Crassulaceae) leaf aqueous extract.

    PubMed

    Ojewole, John A O

    2005-05-13

    In order to scientifically appraise some of the ethnomedical uses of Bryophyllum pinnatum leaves, the present study was undertaken to investigate the antinociceptive, anti-inflammatory and antidiabetic properties of the plant's leaf aqueous extract in experimental animal models. The antinociceptive effect of the herb's leaf extract was evaluated by the 'hot-plate' and 'acetic acid' test models of pain in mice. The anti-inflammatory and antidiabetic effects of the plant's extract were investigated in rats, using fresh egg albumin-induced pedal (paw) oedema, and streptozotocin (STZ)-induced diabetes mellitus. Diclofenac (DIC, 100 mg/kg) and chlorpropamide (250 mg/kg) were used respectively as reference drugs for comparison. Bryophyllum pinnatum leaf aqueous extract (BPE, 25-800 mg/kg i.p.) produced significant (P<0.05-0.001) antinociceptive effects against thermally- and chemically-induced nociceptive pain stimuli in mice. The plant extract (BPE, 25-800 mg/kg p.o. or i.p.) also significantly (P<0.05-0.001) inhibited fresh egg albumin-induced acute inflammation and caused significant (P<0.05-0.001) hypoglycaemia in rats. The results of this experimental animal study suggest that Bryophyllum pinnatum leaf aqueous extract possesses antinociceptive, anti-inflammatory and hypoglycaemic properties. The different flavonoids, polyphenols, triterpenoids and other chemical constituents of the herb are speculated to account for the observed antinociceptive, anti-inflammatory and antidiabetic properties of the plant.

  19. Reverse Transcriptase Activity in Mature Spermatozoa of Mouse

    PubMed Central

    Giordano, Roberto; Magnano, Anna Rosa; Zaccagnini, Germana; Pittoggi, Carmine; Moscufo, Nicola; Lorenzini, Rodolfo; Spadafora, Corrado

    2000-01-01

    We show here that a reverse transcriptase (RT) activity is present in murine epididymal spermatozoa. Sperm cells incubated with human poliovirus RNA can take up exogenous RNA molecules and internalize them in nuclei. Direct PCR amplification of DNA extracted from RNA-incubated spermatozoa indicate that poliovirus RNA is reverse-transcribed in cDNA fragments. PCR analysis of two-cell embryos shows that poliovirus RNA-challenged spermatozoa transfer retrotranscribed cDNA molecules into eggs during in vitro fertilization. Finally, RT molecules can be visualized on sperm nuclear scaffolds by immunogold electron microscopy. These results, therefore, reveal a novel metabolic function in spermatozoa, which may play a role during early embryonic development. PMID:10725323

  20. Blood vessels segmentation of hatching eggs based on fully convolutional networks

    NASA Astrophysics Data System (ADS)

    Geng, Lei; Qiu, Ling; Wu, Jun; Xiao, Zhitao

    2018-04-01

    FCN, trained end-to-end, pixels-to-pixels, predict result of each pixel. It has been widely used for semantic segmentation. In order to realize the blood vessels segmentation of hatching eggs, a method based on FCN is proposed in this paper. The training datasets are composed of patches extracted from very few images to augment data. The network combines with lower layer and deconvolution to enables precise segmentation. The proposed method frees from the problem that training deep networks need large scale samples. Experimental results on hatching eggs demonstrate that this method can yield more accurate segmentation outputs than previous researches. It provides a convenient reference for fertility detection subsequently.

  1. ROLE OF THE EGG JELLY COAT IN PROTECTING HYLA REGILLA AND BUFO CANORUS EMBRYOS FROM ULTRAVIOLET B RADIATION DURING DEVELOPMENT

    EPA Science Inventory

    Previous studies have suggested that Ultraviolet B (UVB) radiation may play a role in amphibian population declines. Some of these studies also indicate that egg hatching success is unaltered in some species of anurans as a result of UVB exposure. It has been proposed that the eg...

  2. Prostate Cancer Research Training in Health Disparities for Undergraduates (PCaRT)

    DTIC Science & Technology

    2011-03-01

    objective is to use statistical methods to evaluate the role of meat , fish, The role of meat , fish and eggs in prostate cancer risk among African-American...The role of meat , fish, eggs and diary products in prostate cancer risk among African-Americans. (Poster) 5. Robertino Simpson, Fisk University...Navigator Lovell A. Jones, Ph.D. * Research Ethics Dr. C. Freund Hazard Communication Standard Training Mr. D. Powell Blood Borne Pathogen Standard

  3. Egg Laying of Cabbage White Butterfly (Pieris brassicae) on Arabidopsis thaliana Affects Subsequent Performance of the Larvae

    PubMed Central

    Blenn, Beatrice; Drechsler, Navina; Gershenzon, Jonathan; Kunze, Reinhard; Hilker, Monika

    2013-01-01

    Plant resistance to the feeding by herbivorous insects has recently been found to be positively or negatively influenced by prior egg deposition. Here we show how crucial it is to conduct experiments on plant responses to herbivory under conditions that simulate natural insect behaviour. We used a well-studied plant – herbivore system, Arabidopsis thaliana and the cabbage white butterfly Pieris brassicae, testing the effects of naturally laid eggs (rather than egg extracts) and allowing larvae to feed gregariously as they do naturally (rather than placing single larvae on plants). Under natural conditions, newly hatched larvae start feeding on their egg shells before they consume leaf tissue, but access to egg shells had no effect on subsequent larval performance in our experiments. However, young larvae feeding gregariously on leaves previously laden with eggs caused less feeding damage, gained less weight during the first 2 days, and suffered twice as high a mortality until pupation compared to larvae feeding on plants that had never had eggs. The concentration of the major anti-herbivore defences of A. thaliana, the glucosinolates, was not significantly increased by oviposition, but the amount of the most abundant member of this class, 4-methylsulfinylbutyl glucosinolate was 1.8-fold lower in larval-damaged leaves with prior egg deposition compared to damaged leaves that had never had eggs. There were also few significant changes in the transcript levels of glucosinolate metabolic genes, except that egg deposition suppressed the feeding-induced up-regulation of FMOGS-OX2, a gene encoding a flavin monooxygenase involved in the last step of 4-methylsulfinylbutyl glucosinolate biosynthesis. Hence, our study demonstrates that oviposition does increase A. thaliana resistance to feeding by subsequently hatching larvae, but this cannot be attributed simply to changes in glucosinolate content. PMID:23527243

  4. Ovipositional responses of the pulse beetle, Bruchus chinensis (Coleoptera: Bruchidae) to extracts and compounds of Capparis decidua.

    PubMed

    Upadhyay, Ravi K; Rohatgi, Leena; Chaubey, Mukesh K; Jain, Subhash C

    2006-12-27

    Extracts of Capparis decidua stems and flowers showed insecticidal and oviposition inhibitory activities against Bruchus chinensis. The LC50 values of these extracts were found to increase with the increase in the polarity of the extract at different exposure periods. For instance, after 96 h, the LC50 values were found to be 3.619, 7.319, and 10.151 microg for CD1, CD2, and CD3, respectively. Extract CD7 was effective only at higher doses. The toxicity was found to be dose- and time-dependent. The females laid lesser number of eggs, when exposed to sublethal doses of different extracts and pure compounds, as compared to control. The maximum oviposition deterrence index was found for extract CD1 followed in decreasing order by CD2, CD3, and CD7. From extract CD1, two compounds were isolated and characterized as triacontanol (C1) and 2-carboxy-1,1-dimethylpyrrolidine (C2). When the females were exposed to sublethal doses of these compounds, they laid lesser number of eggs as compared to the control. C2 was found to have a slightly greater oviposition inhibition effect than C1. From fraction CD7, one novel compound labeled as CDF1 has been isolated and identified as 6-(1-hydroxy-non-3-enyl)tetrahydropyran-2-one. CDF1 has also shown insecticidal and oviposition inhibitory activities against B. chinensis at low concentrations.

  5. Cross-reactivity of Schistosoma mansoni-Fasciola gigantica influenced by saponins.

    PubMed

    Maghraby, Amany Sayed; Shaker, Kamel H; Gaber, Hanaa M

    2009-01-01

    The aim of the present work was to investigate the Schistosoma mansoni and Fasciola gigantica cross-reactivity between adult worms and egg homogenates of the parasites. Immunoprophylactic effects of crude Schistosoma mansoni worms and egg antigens mixed with or without saponins extracted from Atriplex nummularia were studied followed by challenge with 80 cercariae of Schistosoma mansoni. Our results showed that post 1st immunization with schistosome egg antigens (SEA) there was a significant change (P approximately 0.05) in the IgM levels against Fasciola egg homogenate (FgEH) without saponins. Post 2nd immunization with SEA mixed with saponins the levels of IgM increased significantly (P approximately 0.05) against Fasciola worm homogenate (FgWH) as compared with a non-immunized group. Post 2nd immunization the level of IgG was significantly elevated (P approximately 0.05) by SEA mixed with saponins against FgWH. Post 2nd immunizations with SEA mixed with saponins showed a significant change (P approximately 0.05) in IgG levels against FgEH. These results clearly demonstrated that there is a cross-reactivity between Schistosoma mansoni eggs and Fasciola gigantica worms and eggs. Saponins were found to be immunostimulatory adjuvants in our study.

  6. Cryopreservation of bull semen: Evolution from egg yolk based to soybean based extenders.

    PubMed

    Layek, S S; Mohanty, T K; Kumaresan, A; Parks, J E

    2016-09-01

    Since the inception of bovine semen cryopreservation, egg yolk and milk based extenders have been used to protect sperm from the detrimental effects of cooling and freezing. In recent years, demand for alternatives to conventional commercial extenders has arisen as the risk of introducing exotic diseases through transporting egg yolk based products has been recognized. Egg yolk can also interfere with sperm evaluation and the presence of particulate material in the extender may reduce fertility. Soybeans contain lecithin, a phospholipid fraction that can substitute for high molecular weight lipoprotein and phospholipids from egg yolk and prevent or ameliorate damage to the sperm plasma membrane that occurs during extension, cooling, and cryopreservation. Soy lecithin based extenders have been evaluated for processing and freezing bovine semen, although extender from soybean milk has not been studied as extensively. Commercially available soy lecithin based extenders are used increasingly but remain under scrutiny and are not universally accepted. With these observations in mind, this review is intended to examine effects of conventional cryopreservation procedures, methods of assessment, and potential for developing soybean extract as an acceptable alternative to traditional egg yolk and milk based extenders for bull sperm cryopreservation. Copyright © 2016. Published by Elsevier B.V.

  7. Nonvolatile chemical cues affect host-plant ranking by gravid Polygonia c-album females.

    PubMed

    Mozūraitis, Raimondas; Murtazina, Rushana; Nylin, Sören; Borg-Karlson, Anna-Karin

    2012-01-01

    In a multiple-choice test, the preference of egg-laying Polygonia c-album (comma butterfly) females was studied for oviposition on plants bearing surrogate leaves treated with crude methanol extracts obtained from leaves of seven host-plant species: Humulus lupulus, Urtica dioica, Ulmus glabra, Salix caprea, Ribes nigrum, Corylus avellana, and Betula pubescens. The ranking order of surrogate leaves treated with host-plant extracts corresponded well to that reported on natural foliage, except R. nigrum. Thus, host-plant choice in P. c-album seems to be highly dependent on chemical cues. Moreover, after two subsequent fractionations using reversed-phase chromatography the nonvolatile chemical cues residing in the most polar water-soluble fractions evidently provided sufficient information for egg-laying females to discriminate and rank between the samples of more and less preferred plants, since the ranking in these assays was similar to that for natural foliage or whole methanol extracts, while the physical traits of the surrogate leaves remained uniform.

  8. Chum salmon egg extracts induce upregulation of collagen type I and exert antioxidative effects on human dermal fibroblast cultures

    PubMed Central

    Yoshino, Atsushi; Polouliakh, Natalia; Meguro, Akira; Takeuchi, Masaki; Kawagoe, Tatsukata; Mizuki, Nobuhisa

    2016-01-01

    Components of fish roe possess antioxidant and antiaging activities, making them potentially very beneficial natural resources. Here, we investigated chum salmon eggs (CSEs) as a source of active ingredients, including vitamins, unsaturated fatty acids, and proteins. We incubated human dermal fibroblast cultures for 48 hours with high and low concentrations of CSE extracts and analyzed changes in gene expression. Cells treated with CSE extract showed concentration-dependent upregulation of collagen type I genes and of multiple antioxidative genes, including OXR1, TXNRD1, and PRDX family genes. We further conducted in silico phylogenetic footprinting analysis of promoter regions. These results suggested that transcription factors such as acute myeloid leukemia-1a and cyclic adenosine monophosphate response element-binding protein may be involved in the observed upregulation of antioxidative genes. Our results support the idea that CSEs are strong candidate sources of antioxidant materials and cosmeceutically effective ingredients. PMID:27621603

  9. A minimally invasive method for extraction of sturgeon oocytes

    USGS Publications Warehouse

    Candrl, James S.; Papoulias, Diana M.; Tillitt, Donald E.

    2010-01-01

    Fishery biologists, hatchery personnel, and caviar fishers routinely extract oocytes from sturgeon (Acipenseridae) to determine the stage of maturation by checking egg quality. Typically, oocytes are removed either by inserting a catheter into the oviduct or by making an incision in the body cavity. Both methods can be time-consuming and stressful to the fish. We describe a device to collect mature oocytes from sturgeons quickly and effectively with minimal stress on the fish. The device is made by creating a needle from stainless steel tubing and connecting it to a syringe with polyvinyl chloride tubing. The device is filled with saline solution or water, the needle is inserted into the abdominal wall, and eggs are extracted from the fish. Using this device, an oocyte sample can be collected in less than 30 s. Such sampling leaves a minute wound that heals quickly and does not require suturing. The extractor device can easily be used in the field or hatchery, reduces fish handling time, and minimizes stress.

  10. Expression of phosphatidylcholine biosynthetic enzymes during early embryogenesis in the amphibian Bufo arenarum.

    PubMed

    Fernández-Bussy, Rodrigo; Mouguelar, Valeria; Banchio, Claudia; Coux, Gabriela

    2015-04-01

    In the principal route of phosphatidylcholine (PC) synthesis the regulatory steps are catalysed by CTP:phosphocholine cytidylyltransferase (CCT) and choline kinase (CK). Knock-out mice in Pcyt1a (CCT gene) and Chka1 (CK gene) resulted in preimplantation embryonic lethality, demonstrating the essential role of this pathway. However, there is still a lack of detailed CCT and CK expression analysis during development. The aim of the current work was to study the expression during early development of both enzymes in the external-fertilization vertebrate Bufo arenarum. Reverse transcription polymerase chain reaction (RT-PCR) and western blot confirmed their presence in unfertilized eggs. Analysis performed in total extracts from staged embryos showed constant protein levels of both enzymes until the 32-cell stage: then they decreased, reaching a minimum in the gastrula before starting to recover. CTP:phosphocholine cytidylyltransferase is an amphitropic enzyme that inter-converts between cytosolic inactive and membrane-bound active forms. Immunoblot analysis demonstrated that the cytosolic:total CCT protein ratio does not change throughout embryogenesis, suggesting a progressive decline of CCT activity in early development. However, PC (and phosphatidylethanolamine) content per egg/embryo remained constant throughout the stages analysed. In conclusion, the current data for B. arenarum suggest that net synthesis of PC mediated by CCT and CK is not required in early development and that supplies for membrane biosynthesis are fulfilled by lipids already present in the egg/embryo reservoirs.

  11. Egg-Citing! Isolation of Protoporphyrin IX from Brown Eggshells and Its Detection by Optical Spectroscopy and Chemiluminescence

    ERIC Educational Resources Information Center

    Dean, Michelle L.; Miller, Tyson A.; Bruckner, Christian

    2011-01-01

    A simple and cost-effective laboratory experiment is described that extracts protoporphyrin IX from brown eggshells. The porphyrin is characterized by UV-vis and fluorescence spectroscopy. A chemiluminescence reaction (peroxyoxalate ester fragmentation) is performed that emits light in the UV region. When the porphyrin extract is added as a fluor…

  12. Proteome changes in rainbow trout (Oncorhynchus mykiss) fertilized eggs as an effect of triploidization heat-shock treatment.

    PubMed

    Babaheydari, Samad Bahrami; Keyvanshokooh, Saeed; Dorafshan, Salar; Johari, Seyed Ali

    2016-03-01

    The aim of the present study was to explore proteome changes in rainbow trout (Oncorhynchus mykiss) fertilized eggs as an effect of triploidization heat-shock treatment. Eggs and milt were taken from eight females and six males. The gametes were pooled to minimize the individual differences. After insemination, the eggs were incubated at 10°C for 10min. Half of the fertilized eggs were then subjected to heat shock for 10min submerged in a 28°C water bath to induce triploidy. The remainder were incubated normally and used as diploid controls. Three batches of eggs were randomly selected from each group and were incubated at 10-11°C under the same environmental conditions in hatchery troughs until the fry stage. Triplicate samples of 30 eggs (10 eggs per trough) from each group were randomly selected 1.5h post-fertilization for proteome extraction. Egg proteins were analyzed using two-dimensional electrophoresis (2-DE) and MALDI-TOF/TOF mass spectrometry. Based on the results from the statistical analyses, 15 protein spots were found to decrease significantly in abundance in heat-shock treated group and were selected for identification. Out of 15 protein spots showing altered abundance, 14 spots were successfully identified. All of the egg proteins identified in our study were related to vitellogenin (vtg). Decreased abundance of vitellogenin in heat-shock treated eggs in our study may either be explained by (i) higher utilization of vtg as an effect of increased cell size in triploids or (ii) changed metabolism in response to heat-shock stress and (iii) diffusion of vtg through chorion due to incidence of egg shell damage. Decreased abundance of vitellogenin in heat-shock treated eggs was associated with reduced early survival rates and lowered growth performance of triploid fish. Copyright © 2016 Elsevier B.V. All rights reserved.

  13. The role of egg-nest contrast in the rejection of brood parasitic eggs.

    PubMed

    Aidala, Zachary; Croston, Rebecca; Schwartz, Jessica; Tong, Lainga; Hauber, Mark E

    2015-04-15

    Hosts of avian brood parasites can avoid the reproductive costs of raising genetically unrelated offspring by rejecting parasitic eggs. The perceptual cues and controls mediating parasitic egg discrimination and ejection are well studied: hosts are thought to use differences in egg color, brightness, maculation, size and shape to discriminate between their own and foreign eggs. Most theories of brood parasitism implicitly assume that the primary criteria to which hosts attend when discriminating eggs are differences between the eggs themselves. However, this assumption is confounded by the degree to which chromatic and achromatic characteristics of the nest lining co-vary with egg coloration, so that egg-nest contrast per se might be the recognition cue driving parasitic egg detection. Here, we systematically tested whether and how egg-nest contrast itself contributes to foreign egg discrimination. In an artificial parasitism experiment, we independently manipulated egg color and nest lining color of the egg-ejector American robin (Turdus migratorius), a host of the obligate brood parasitic brown-headed cowbird (Molothrus ater). We hypothesized that the degree of contrast between foreign eggs and the nest background would affect host egg rejection behavior. We predicted that experimentally decreasing egg-nest chromatic and achromatic contrast (i.e. rendering parasitic eggs more cryptic against the nest lining) would decrease rejection rates, while increasing egg-nest contrast would increase rejection rates. In contrast to our predictions, egg-nest contrast was not a significant predictor of egg ejection patterns. Instead, egg color significantly predicted responses to parasitism. We conclude that egg-egg differences are the primary drivers of egg rejection in this system. Future studies should test for the effects of egg-nest contrast per se in predicting parasitic egg recognition in other host-parasite systems, including those hosts building enclosed nests and those parasites laying cryptic eggs, as an alternative to hypothesized effects of egg-egg contrast. © 2015. Published by The Company of Biologists Ltd.

  14. Ovicidal, larvicidal and adulticidal properties of Asparagus racemosus (Willd.) (Family: Asparagaceae) root extracts against filariasis (Culex quinquefasciatus), dengue (Aedes aegypti) and malaria (Anopheles stephensi) vector mosquitoes (Diptera: Culicidae).

    PubMed

    Govindarajan, Marimuthu; Sivakumar, Rajamohan

    2014-04-01

    Several diseases are associated to the mosquito-human interaction. Mosquitoes are the carriers of severe and well-known illnesses such as malaria, arboviral encephalitis, dengue fever, chikungunya fever, West Nile virus and yellow fever. These diseases produce significant morbidity and mortality in humans and livestock around the world. The present investigation was undertaken to study the ovicidal, larvicidal and adulticidal activities of crude hexane, ethyl acetate, benzene, chloroform and methanol extracts of root of Asparagus racemosus were assayed for their toxicity against three important vector mosquitoes, viz., Culex quinquefasciatus, Aedes aegypti and Anopheles stephensi (Diptera: Culicidae). The mean percent hatchability of the eggs was observed after 48 h post-treatment. The percent hatchability was inversely proportional to the concentration of extract and directly proportional to the eggs. All the five solvent extracts showed moderate ovicidal activity; however, the methanol extract showed the highest ovicidal activity. The methanol extract of Asparagus racemosus against Culex quinquefasciatus, Aedes aegypti and Anopheles stephensi exerted 100% mortality (zero hatchability) at 375, 300 and 225 ppm, respectively. Control eggs showed 99-100% hatchability. The larval mortality was observed after 24 h of exposure. All extracts showed moderate larvicidal effects; however, the highest larval mortality was found in methanol extract of root of Asparagus racemosus against the larvae of Culex quinquefasciatus, Aedes aegypti and Anopheles stephensi with the LC50 and LC90 values were 115.13, 97.71 and 90.97 ppm and 210.96, 179.92, and 168.82 ppm, respectively. The adult mortality was observed after 24 h recovery period. The plant crude extracts showed dose-dependent mortality. At higher concentrations, the adult showed restless movement for some times with abnormal wagging and then died. Among the extracts tested, the highest adulticidal activity was observed in methanol extract against Anopheles stephensi followed by Aedes aegypti and Culex quinquefasciatus with the LD50 and LD90 values were 120.44, 135.60, and 157.71 ppm and 214.65, 248.35, and 290.95 ppm, respectively. No mortality was recorded in the control. The finding of the present investigation revealed that the root extract of Asparagus racemosus possess remarkable ovicidal, larvicidal and adulticidal activity against medically important vector mosquitoes and this is the low cost and ideal eco-friendly approach for the control of mosquitoes. This is the first report on the mosquito ovicidal, larvicidal and adulticidal activities of the reported Asparagus racemosus root.

  15. Fate of egg proteins during the development of Columba livia domestica embryo.

    PubMed

    Shbailat, Seba Jamal; Aslan, Ibtisam Omar

    2018-01-01

    The transfer of egg white into the yolk and consumption of yolk proteins by the embryo are largely unexplored in the pigeon Columba livia domestica. Here, we investigated the route of egg white transfer as well as the degradation and uptake of yolk proteins by the pigeon embryo. Initially, we tested the electrophoretic patterns of proteins in different egg compartments throughout development. Then, we used lysozyme as a reference protein to follow the egg white transfer, and we measured its activity using Micrococcus lysodeikticus as a substrate. Moreover, we determined the general protease activity during different developmental stages in the yolk using casein. Finally, we examined the expression of aminopeptidase-N (APN) and oligopeptide transporter PepT1 genes in the yolk sac membrane (YSM) from incubation day 8 until day 17. Several electrophoretic bands of presumptive egg white proteins appeared in different egg compartments. Also, lysozyme activity was detected chronologically in the egg compartments. It appeared on day 12 in the amniotic and intestinal fluids and on day 14 in the yolk. Moreover, protease activity in the yolk increased significantly on day 14 and thereafter. APN expression was largest on day 8 and reduced generally afterward, whereas PepT1 expression peaked between days 13 and 15 but then reduced substantially. Our results suggest that the egg white proteins move through the amnion and intestine into the yolk where they undergo degradation by the activated proteases. Furthermore, the YSM appears to have a role in protein consumption, and this role decreases toward hatch. © 2018 Wiley Periodicals, Inc.

  16. Hen Egg as an Antioxidant Food Commodity: A Review

    PubMed Central

    Nimalaratne, Chamila; Wu, Jianping

    2015-01-01

    Intake of antioxidants through diet is known to be important in reducing oxidative damage in cells and improving human health. Although eggs are known for their exceptional, nutritional quality, they are not generally considered as antioxidant foods. This review aims to establish the importance of eggs as an antioxidant food by summarizing the current knowledge on egg-derived antioxidants. Eggs have various natural occurring compounds including the proteins ovalbumin, ovotransferrin and lysozyme in egg white, as well as phosvitin, carotenoids and free aromatic amino acids in egg yolk. Some lipophilic antioxidants such as vitamin E, carotenoids, selenium, iodine and others can be transferred from feed into egg yolk to produce antioxidant-enriched eggs. The bioactivity of egg antioxidants can be affected by food processing, storage and gastrointestinal digestion. Generally thermal processing methods can promote loss of antioxidant properties in eggs due to oxidation and degradation, whereas gastrointestinal digestion enhances the antioxidant properties, due to the formation of new antioxidants (free amino acids and peptides). In summary, in addition to its well-known nutritional contribution to our diet, this review emphasizes the role of eggs as an important antioxidant food. PMID:26404361

  17. Larvicidal, ovicidal and pupicidal activities of Gliricidia sepium (Jacq.) (Leguminosae) against the malarial vector, Anopheles stephensi Liston (Culicidae: Diptera).

    PubMed

    Krishnappa, Kaliyamoorthy; Dhanasekaran, Shanmugam; Elumalai, Kuppusamy

    2012-08-01

    To investigate the potentiality of mosquitocidal activity of Gliricidia sepium (G. sepium) (Jacq.) (Leguminosae). Twenty five early third instar larvae of Anopheles stephensi (An. stephensi) were exposed to various concentrations (50-250 ppm) and the 24 h LC(50) values of the G. sepium extract was determined by probit analysis. The ovicidal activity was determined against An. stephensi to various concentrations ranging from 25-100 ppm under laboratory conditions. The eggs hatchability was assessed 48 h post treatment. The pupicidal activity was determined against An. stephensi to various concentrations ranging from 25-100 ppm. Mortality of each pupa was recorded after 24 h of exposure to the extract. Results pertaining to the experiment clearly revealed that ethanol extract showed significant larvicidal, ovicidal and pupicidal activity against the An. stephensi. Larvicidal activity of ethanol extracts of G. sepium showed maximum mortality in 250 ppm concentration (96.0±2.4)%. Furthermore, the LC(50) was found to be 121.79 and the LC(90) value was recorded to be 231.98 ppm. Ovicidal activity of ethanol extract was assessed by assessing the egg hatchability. Highest concentration of both solvent extracts exhibited 100% ovicidal activity. Similarly, pupae exposed to different concentrations of ethanol extract were found dead with 58.10% adult emergence when it was treated with 25 ppm concentration. Similarly, 18.36 (n=30; 61.20%); 21.28(70.93) and 27.33(91.10) pupal mortality was recorded from the experimental pupae treated with 50, 75 and 100 ppm concentration of extracts. Three fractions have been tested for their larvicidal activity of which the Fraction 3 showed the LC(50) and LC(90) values of 23.23 and 40.39 ppm. With regard to the ovicidal effect fraction 3 showed highest ovicidal activities than the other two fractions. Furthermore, there were no hatchability was recorded above 50 ppm (100% egg mortality) in the experimental group. Statistically significant pupicidal activity was recorded from 75 ppm concentration. From the results it can be concluded the crude extract of G. sepium is an excellent potential for controlling An. stephensi mosquito. It is apparent that, fraction 3 possess a novel and active principle which could be responsible for those biological activities. Copyright © 2012 Hainan Medical College. Published by Elsevier B.V. All rights reserved.

  18. Anti-helminthic activity of Momordica charantia L. against Fasciola hepatica eggs after twelve days of incubation in vitro.

    PubMed

    Pereira, Cíntia A J; Oliveira, Laura L S; Coaglio, Aytube L; Santos, Fernanda S O; Cezar, Rodolfo S M; Mendes, Tiago; Oliveira, Fernando L P; Conzensa, Gustavo; Lima, Walter S

    2016-09-15

    Fasciolosis, a parasitic disease caused by the trematode Fasciola hepatica underreported is expanding both in human and animal population, throughout the world. The constant use of synthetic drugs to treat this condition has led to the natural selection of resistant strains of the parasite. Hence, there is a growing focus on the potential anti-helminthic properties of medicinal plants and phytopharmaceuticals. The current study assessed the potential anti-fasciolicide action of Momordica charantia leaf extracts and fractions on the eggs of F. hepatica parasites. The lyophilized crude extract (CE) of M. charantia leaves and its sub-fractions, obtained from liquid-liquid partitioning with organic solvents, were analysed by High Performance Liquid Chromatography (HPLC), suspended in 1% DMSO and used in in vitro tests. Quadruplicates of 50F. hepatica eggs were incubated at 23°C with M. charantia leaf CE in different concentrations. After 12days no larvae were formed in eggs incubated with CE concentrations above 12.5mg/mL. Eggs incubated with CE sub-fractions at concentrations of 1000, 100, 10, 1, 0.1, 0.01μg/mL affected embryonic development, with n-butanol presenting the strongest inhibition of miracidia formation. In contrast, on the 12th day, 90% of the miracidia hatched in the control experiments using 0.03% DMSO whereas embryogenesis was completely abolished with any concentration of albendazole sulphoxide ABZ(SO). Chemical analysis of the CE and sub-fractions revealed a prominent presence of flavonoids. HPLC-MS confirmed Quercetin to be one of the main flavonoids present in the CE and the n-butanol subfraction. This is the first study to analyse the potential anti-fasciolicide action of M. charantia leaf CE and subfractions. Copyright © 2016 Elsevier B.V. All rights reserved.

  19. Protein profiles of hatchery egg shell membrane.

    PubMed

    Rath, N C; Liyanage, R; Makkar, S K; Lay, J O

    2016-01-01

    Eggshells which consist largely of calcareous outer shell and shell membranes, constitute a significant part of poultry hatchery waste. The shell membranes (ESM) not only contain proteins that originate from egg whites but also from the developing embryos and different contaminants of microbial and environmental origins. As feed supplements, during post hatch growth, the hatchery egg shell membranes (HESM) have shown potential for imparting resistance of chickens to endotoxin stress and exert positive health effects. Considering that these effects are mediated by the bioactive proteins and peptides present in the membrane, the objective of the study was to identify the protein profiles of hatchery eggshell membranes (HESM). Hatchery egg shell membranes were extracted with acidified methanol and a guanidine hydrochloride buffer then subjected to reduction/alkylation, and trypsin digestion. The methanol extract was additionally analyzed by matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF-MS). The tryptic digests were analyzed by liquid chromatography and tandem mass spectrometry (LC-MS-MS) to identify the proteins. Our results showed the presence of several proteins that are inherent and abundant in egg white such as, ovalbumin, ovotransferrin, ovocleidin-116, and lysozyme, and several proteins associated with cytoskeletal, cell signaling, antimicrobial, and catalytic functions involving carbohydrate, nucleic acid, and protein metabolisms. There were some blood derived proteins most likely originating from the embryos and several other proteins identified with different aerobic, anaerobic, gram positive, gram negative, soil, and marine bacterial species some commensals and others zoonotic. The variety of bioactive proteins, particularly the cell signaling and enzymatic proteins along with the diverse microbial proteins, make the HESM suitable for nutritional and biological application to improve post hatch immunity of poultry.

  20. The multivariate egg: quantifying within- and among-clutch correlations between maternally derived yolk immunoglobulins and yolk androgens using multivariate mixed models.

    PubMed

    Postma, Erik; Siitari, Heli; Schwabl, Hubert; Richner, Heinz; Tschirren, Barbara

    2014-03-01

    Egg components are important mediators of prenatal maternal effects in birds and other oviparous species. Because different egg components can have opposite effects on offspring phenotype, selection is expected to favour their mutual adjustment, resulting in a significant covariation between egg components within and/or among clutches. Here we tested for such correlations between maternally derived yolk immunoglobulins and yolk androgens in great tit (Parus major) eggs using a multivariate mixed-model approach. We found no association between yolk immunoglobulins and yolk androgens within clutches, indicating that within clutches the two egg components are deposited independently. Across clutches, however, there was a significant negative relationship between yolk immunoglobulins and yolk androgens, suggesting that selection has co-adjusted their deposition. Furthermore, an experimental manipulation of ectoparasite load affected patterns of covariance among egg components. Yolk immunoglobulins are known to play an important role in nestling immune defence shortly after hatching, whereas yolk androgens, although having growth-enhancing effects under many environmental conditions, can be immunosuppressive. We therefore speculate that variation in the risk of parasitism may play an important role in shaping optimal egg composition and may lead to the observed pattern of yolk immunoglobulin and yolk androgen deposition across clutches. More generally, our case study exemplifies how multivariate mixed-model methodology presents a flexible tool to not only quantify, but also test patterns of (co)variation across different organisational levels and environments, allowing for powerful hypothesis testing in ecophysiology.

  1. The Role of Baked Egg and Milk in the Diets of Allergic Children.

    PubMed

    Robinson, Melissa L; Lanser, Bruce J

    2018-02-01

    Baked egg and baked milk are tolerated by most children who are allergic to hen's egg and cow's milk. Incorporating baked goods into the diets of allergic children may help them outgrow their primary allergy more quickly, with changes observed akin to immunotherapy. Benefits may also include increased quality of life and improved nutritional status. The search for a reliable biomarker to predict tolerance to baked goods is ongoing. Most children with a milk or egg allergy who are not previously tolerating egg or milk in baked goods should be offered an observed oral food challenge. Copyright © 2017 Elsevier Inc. All rights reserved.

  2. Variation in incubation periods and egg metabolism in mallards: Intrinsic mechanisms to promote hatch synchrony

    USGS Publications Warehouse

    MacCluskie, Margaret C.; Flint, Paul L.; Sedinger, James S.

    1997-01-01

    We investigated factors affecting incubation time and metabolic rates of Mallard (Anas platyrhynchos) eggs incubated under constant environmental conditions. Time required to reach the star-pipped stage of hatch varied significantly among females, but not with laying sequence or egg size. Metabolic rate of eggs varied positively with position in the laying sequence and tended to vary among females. Metabolic rate did not vary with egg volume or incubation length. Our results indicate metabolic rate may act as one synchronization mechanism for hatch. The role of maternal effects in development time should be considered in subsequent studies of incubation time in ducks.

  3. In vitro efficacy of essential oils and extracts of Schinus molle L. against Ctenocephalides felis felis.

    PubMed

    Batista, Lilian C De S O; Cid, Yara P; De Almeida, Ana Paula; Prudêncio, Edlene R; Riger, Cristiano J; De Souza, Marco A A; Coumendouros, Katherine; Chaves, Douglas S A

    2016-04-01

    Extracts and essential oils from plants are important natural sources of pesticides. These compounds are considered an alternative to control ectoparasites of veterinary importance. Schinus molle, an endemic species of Brazil, produces a high level of essential oil and several other compounds. The aim of this work was to determinate the chemical composition of extracts and essential oils of S. molle and further to evaluate the activity against eggs and adults of Ctenocephalides felis felis, a predominant flea that infests dogs and cats in Brazil. In an in vitro assay, the non-polar (n-hexane) extract showed 100% efficacy (800 µg cm(-2); LD50 = 524·80 µg cm(-2)) at 24 and 48 h. Its major compound was lupenone (50·25%). Essential oils from fruits and leaves were evaluated, and had 100% efficacy against adult fleas at 800 µg cm(-2) (LD50 = 353·95 µg cm(-2)) and at 50 µg cm(-2) (LD50 = 12·02 µg cm(-2)), respectively. On the other hand, the essential oil from fruits and leaves was not active against flea eggs. This is the first study that reports the insecticidal effects of essential oils and extracts obtained from Schinus molle against Ctenocephalides felis felis.

  4. Aflatoxin B1 in eggs and chicken livers by dispersive liquid-liquid microextraction and HPLC.

    PubMed

    Amirkhizi, Behzad; Arefhosseini, Seyed Rafie; Ansarin, Masoud; Nemati, Mahboob

    2015-01-01

    A rapid, low-cost and simple technique has been developed for the determination of aflatoxin B1 (AFB1) in eggs and livers using high-performance liquid chromatography (HPLC) with UV detection. In this study, the presence of AFB1 was investigated in 150 eggs and 50 chicken livers from the local market of Tabriz, Iran. AFB1 was extracted with a mixture of acetonitrile:water (80:20) and cleaned up by dispersive liquid-liquid microextraction which is a very economical, fast and sensitive method. AFB1 was quantified by HPLC-UV without need for any complex derivatisation in samples to enhance the detection. The results showed that 72% of the liver and 58% of the egg samples were contaminated with AFB1 ranging from 0.30 to 16.36 µg kg (̶1). limit of detection and limit of quantification for AFB1 were 0.08 and 0.28 µg kg (̶ 1), respectively. The proposed method is suitable for fast analysing of AFB1 in egg and liver samples.

  5. [Investigation of Coxiella burnetii contamination in commercial milk and PCR method for the detection of C. burnetii in egg].

    PubMed

    Hirai, Akihiko; Kaneko, Seiji; Nakama, Akiko; Ishizaki, Naoto; Odagiri, Megumi; Kai, Akemi; Sadamasu, Kenji; Shinkai, Takayuki; Yano, Kazuyoshi; Morozumi, Satoshi

    2005-06-01

    A total of 244 milk samples collected from supermarkets in Tokyo were examined for contamination with Coxiella burnetii. C. burnetii DNA was detected in 131 (53.7%) of the samples by nested PCR. PCR-positive samples were injected into immunosuppressed A/J strain mice. Of the 22 PCR-positive milk samples tested, none resulted in isolation of C. burnetii from the mice. Heat-treatment was sufficient to inactivate C. burnetii in commercial milk. In addition, a PCR detection method for C. burnetii in chicken egg was developed. Egg yolk was added to an equal volume of 1 mol/L of NaCl phosphate buffer and homogenized for removal of protein and lipid. After centrifugal separation, the supernatant was removed, and template DNA in the precipitate was extracted using SDS, proteinase K and NaI. Using such prepared samples, 3.2 x 10(1) C. burnetii particles in 1 g of egg yolk could be detected by nested PCR. All of 200 chicken egg samples collected from supermarkets in Tokyo were negative for C. burnetii by the nested PCR method.

  6. Bipolarization and Poleward Flux Correlate during Xenopus Extract Spindle AssemblyV⃞

    PubMed Central

    Mitchison, T.J.; Maddox, P.; Groen, A.; Cameron, L.; Perlman, Z.; Ohi, R.; Desai, A.; Salmon, E.D.; Kapoor, T.M.

    2004-01-01

    We investigated the mechanism by which meiotic spindles become bipolar and the correlation between bipolarity and poleward flux, using Xenopus egg extracts. By speckle microscopy and computational alignment, we find that monopolar sperm asters do not show evidence for flux, partially contradicting previous work. We account for the discrepancy by describing spontaneous bipolarization of sperm asters that was missed previously. During spontaneous bipolarization, onset of flux correlated with onset of bipolarity, implying that antiparallel microtubule organization may be required for flux. Using a probe for TPX2 in addition to tubulin, we describe two pathways that lead to spontaneous bipolarization, new pole assembly near chromatin, and pole splitting. By inhibiting the Ran pathway with excess importin-alpha, we establish a role for chromatin-derived, antiparallel overlap bundles in generating the sliding force for flux, and we examine these bundles by electron microscopy. Our results highlight the importance of two processes, chromatin-initiated microtubule nucleation, and sliding forces generated between antiparallel microtubules, in self-organization of spindle bipolarity and poleward flux. PMID:15385629

  7. Probing the biology of cell boundary conditions through confinement of Xenopus cell-free cytoplasmic extracts.

    PubMed

    Bermudez, Jessica G; Chen, Hui; Einstein, Lily C; Good, Matthew C

    2017-01-01

    Cell-free cytoplasmic extracts prepared from Xenopus eggs and embryos have for decades provided a biochemical system with which to interrogate complex cell biological processes in vitro. Recently, the application of microfabrication and microfluidic strategies in biology has narrowed the gap between in vitro and in vivo studies by enabling formation of cell-size compartments containing functional cytoplasm. These approaches provide numerous advantages over traditional biochemical experiments performed in a test tube. Most notably, the cell-free cytoplasm is confined using a two- or three-dimensional boundary, which mimics the natural configuration of a cell. This strategy enables characterization of the spatial organization of a cell, and the role that boundaries play in regulating intracellular assembly and function. In this review, we describe the marriage of Xenopus cell-free cytoplasm and confinement technologies to generate synthetic cell-like systems, the recent biological insights they have enabled, and the promise they hold for future scientific discovery. © 2017 Wiley Periodicals, Inc.

  8. Plants and insect eggs: how do they affect each other?

    PubMed

    Hilker, Monika; Meiners, Torsten

    2011-09-01

    Plant-insect interactions are not just influenced by interactions between plants and the actively feeding stages, but also by the close relationships between plants and insect eggs. Here, we review both effects of plants on insect eggs and, vice versa, effects of eggs on plants. We consider the influence of plants on the production of insect eggs and address the role of phytochemicals for the biosynthesis and release of insect sex pheromones, as well as for insect fecundity. Effects of plants on insect oviposition by contact and olfactory plant cues are summarised. In addition, we consider how the leaf boundary layer influences both insect egg deposition behaviour and development of the embryo inside the egg. The effects of eggs on plants involve egg-induced changes of photosynthetic activity and of the plant's secondary metabolism. Except for gall-inducing insects, egg-induced changes of phytochemistry were so far found to be detrimental to the eggs. Egg deposition can induce hypersensitive-like plant response, formation of neoplasms or production of ovicidal plant substances; these plant responses directly harm the eggs. In addition, egg deposition can induce a change of the plant's odour and leaf surface chemistry which serve indirect plant defence with the help of antagonists of the insect eggs. These egg-induced changes lead to attraction of egg parasitoids and their arrestance on a leaf, respectively. Finally, we summarise knowledge of the elicitors of egg-induced plant changes and address egg-induced effects on the plant's transcriptional pattern. Copyright © 2011 Elsevier Ltd. All rights reserved.

  9. DIETARY BAKED EGG ACCELERATES RESOLUTION OF EGG ALLERGY IN CHILDREN

    PubMed Central

    Leonard, Stephanie A.; Sampson, Hugh A.; Sicherer, Scott H.; Noone, Sally; Moshier, Erin L.; Godbold, James; Nowak-Wȩgrzyn, Anna

    2012-01-01

    Background Baked egg is tolerated by a majority of egg-allergic children. Objective To characterize immunologic changes associated with ingestion of baked egg and evaluate the role that baked egg diets plays in the development of tolerance to regular egg. Methods Egg-allergic subjects who tolerated baked egg challenge incorporated baked egg into their diet. Immunologic parameters were measured at follow-up visits. A comparison group strictly avoiding egg was used to evaluate the natural history of the development of tolerance. Results Of the 79 subjects in the intent-to-treat group followed for a median of 37.8 months, 89% now tolerate baked egg and 53% now tolerate regular egg. Of 23 initial baked egg-reactive subjects, 14 (61%) subsequently tolerated baked egg and 6 (26%) now tolerate regular egg. Within the initially baked egg-reactive group, subjects with persistent reactivity to baked egg had higher median baseline egg white (EW)-specific IgE levels (13.5 kUA/L) than those who subsequently tolerated baked egg (4.4 kUA/L; P=0.04) and regular egg (3.1 kUA/L, P=0.05). In subjects ingesting baked egg, EW-induced SPT wheal diameter and EW-, ovalbumin-, and ovomucoid-specific IgE levels decreased significantly, while ovalbumin- and ovomucoid-specific IgG4 levels increased significantly. Subjects in the per-protocol group were 14.6 times more likely to develop regular egg tolerance than subjects in the comparison group (P < 0.0001), and they developed tolerance earlier (median 50.0 versus 78.7 months; P<0.0001). Conclusion Initiation of a baked egg diet accelerates the development of regular egg tolerance compared to strict avoidance. Higher serum EW-specific IgE level is associated with persistent baked and regular egg reactivity, while initial baked egg reactivity is not. PMID:22846751

  10. Dietary baked egg accelerates resolution of egg allergy in children.

    PubMed

    Leonard, Stephanie A; Sampson, Hugh A; Sicherer, Scott H; Noone, Sally; Moshier, Erin L; Godbold, James; Nowak-Węgrzyn, Anna

    2012-08-01

    Baked egg is tolerated by a majority of egg-allergic children. To characterize immunologic changes associated with ingestion of baked egg and evaluate the role that baked egg diets play in the development of tolerance to regular egg. Egg-allergic subjects who tolerated baked egg challenge incorporated baked egg into their diet. Immunologic parameters were measured at follow-up visits. A comparison group strictly avoiding egg was used to evaluate the natural history of the development of tolerance. Of the 79 subjects in the intent-to-treat group followed for a median of 37.8 months, 89% now tolerate baked egg and 53% now tolerate regular egg. Of 23 initially baked egg-reactive subjects, 14 (61%) subsequently tolerated baked egg and 6 (26%) now tolerate regular egg. Within the initially baked egg-reactive group, subjects with persistent reactivity to baked egg had higher median baseline egg white (EW)-specific IgE levels (13.5 kU(A)/L) than those who subsequently tolerated baked egg (4.4 kU(A)/L; P= .04) and regular egg (3.1 kU(A)/L; P= .05). In subjects ingesting baked egg, EW-induced skin prick test wheal diameter and EW-, ovalbumin-, and ovomucoid-specific IgE levels decreased significantly, while ovalbumin- and ovomucoid-specific IgG(4) levels increased significantly. Subjects in the per-protocol group were 14.6 times more likely than subjects in the comparison group (P< .0001) to develop regular egg tolerance, and they developed tolerance earlier (median 50.0 vs 78.7 months; P< .0001). Initiation of a baked egg diet accelerates the development of regular egg tolerance compared with strict avoidance. Higher serum EW-specific IgE level is associated with persistent baked and regular egg reactivity, while initial baked egg reactivity is not. Copyright © 2012 American Academy of Allergy, Asthma & Immunology. Published by Mosby, Inc. All rights reserved.

  11. Does contrast between eggshell ground and spot coloration affect egg rejection?

    PubMed

    Dainson, Miri; Hauber, Mark E; López, Analía V; Grim, Tomáš; Hanley, Daniel

    2017-08-01

    Obligate avian brood parasitic species impose the costs of incubating foreign eggs and raising young upon their unrelated hosts. The most common host defence is the rejection of parasitic eggs from the nest. Both egg colours and spot patterns influence egg rejection decisions in many host species, yet no studies have explicitly examined the role of variation in spot coloration. We studied the American robin Turdus migratorius, a blue-green unspotted egg-laying host of the brown-headed cowbird Molothrus ater, a brood parasite that lays non-mimetic spotted eggs. We examined host responses to model eggs with variable spot coloration against a constant robin-mimetic ground colour to identify patterns of rejection associated with perceived contrast between spot and ground colours. By using avian visual modelling, we found that robins were more likely to reject eggs whose spots had greater chromatic (hue) but not achromatic (brightness) contrast. Therefore, egg rejection decision rules in the American robin may depend on the colour contrast between parasite eggshell spot and host ground coloration. Our study also suggests that egg recognition in relation to spot coloration, like ground colour recognition, is tuned to the natural variation of avian eggshell spot colours but not to unnatural spot colours.

  12. EVALUATION OF SUBLETHAL EFFECTS OF Ipomoea cairica LINN. EXTRACT ON LIFE HISTORY TRAITS OF DENGUE VECTORS

    PubMed Central

    ZUHARAH, Wan Fatma; AHBIRAMI, Rattanam; DIENG, Hamady; THIAGALETCHUMI, Maniam; FADZLY, Nik

    2016-01-01

    Plant derived insecticides have considerable potential for mosquito control because these products are safer than conventional insecticides. This study aimed to investigate sublethal activities of Ipomoea carica or railway creeper crude acethonilic extract against life history trait of dengue vectors, Aedes albopictus and Aedes aegypti. The late third instar larvae of Ae. albopictus and Ae. aegypti were exposed to a sublethal dose at LC50 and larvae that survived were further cultured. Overall, Ipomea cairica crude extracts affected the whole life history of both Aedes species. The study demonstrated significantly lower egg production (fecundity) and eggs hatchability (fertility) in Ae. albopictus. The sublethal dose of crude extracts reduced significantly the width of larval head capsule and the wing length of both sexes in both Aedes species. The significance of sublethal effects of I. cairica against Aedes mosquitoes was an additional hallmark to demonstrate further activity of this plant despite its direct toxicity to the larvae. The reduced reproductive capacity as well as morphological and physiological anomalies are some of the effects that make I. cairica a potential candidate to be used as a new plant-based insecticide to control dengue vectors. PMID:27253746

  13. Both cyclin A and cyclin E have S-phase promoting (SPF) activity in Xenopus egg extracts.

    PubMed

    Strausfeld, U P; Howell, M; Descombes, P; Chevalier, S; Rempel, R E; Adamczewski, J; Maller, J L; Hunt, T; Blow, J J

    1996-06-01

    Extracts of activated Xenopus eggs in which protein synthesis has been inhibited support a single round of chromosomal DNA replication. Affinity-depletion of cyclin dependent kinases (Cdks) from these extracts blocks the initiation of DNA replication. We define 'S-phase promoting factor' (SPF) as the Cdk activity required for DNA replication in these Cdk-depleted extracts. Recombinant cyclins A and E, but not cyclin B, showed significant SPF activity. High concentrations of cyclin A promoted entry into mitosis, which inhibited DNA replication. In contrast, high concentrations of cyclin E1 promoted neither nuclear envelope disassembly nor full chromosome condensation. In the early embryo cyclin E1 complexes exclusively with Cdk2 and cyclin A is complexed predominantly with Cdc2; only later in development does cyclin A associate with Cdk2. We show that baculovirus-produced complexes of cyclin A-Cd2, cyclin A-Cdk2 and cyclin E-Cdk2 could each provide SPF activity. These results suggest that although in the early Xenopus embryo cyclin E1-Cdk2 is sufficient to support entry into S-phase, cyclin A-Cdc2 provides a significant additional quantity of SPF as its levels rise during S phase.

  14. Role of Caspase-3 Cleaved IP3R1 on Ca2+ Homeostasis and Developmental Competence of Mouse Oocytes and Eggs

    PubMed Central

    Zhang, Nan; Fissore, Rafael. A.

    2014-01-01

    Apoptosis in most cell types is accompanied by altered Ca2+ homeostasis. During apoptosis, caspase-3 mediated cleavage of the type 1 inositol 1,4,5-trisphosphate receptor (IP3R1) generates a 95-kDa C-terminal fragment (C-IP3R1), which represents the channel domain of the receptor. Aged mouse eggs display abnormal Ca2+ homeostasis and express C-IP3R1, although whether or not C-IP3R1 expression contributes to Ca2+ misregulation or a decrease in developmental competency is unknown. We sought to answer these questions by injecting in mouse oocytes and eggs cRNAs encoding CIP3R1. We found that: 1) expression of C-IP3R1 in eggs lowered the Ca2+ content of the endoplasmic reticulum (ER), although, as C-IP3R1 is quickly degraded at this stage, its expression did not impair pre-implantation embryo development; 2) expression of CIP3R1 in eggs enhanced fragmentation associated with aging; 3) endogenous IP3R1 is required for aging associated apoptosis, as its down-regulation prevented fragmentation, and expression of C-IP3R1 in eggs with downregulated IP3R1 partly restored fragmentation; 4) C-IP3R1 expression in GV oocytes resulted in persistent levels of protein, which abolished the increase in the ER releasable Ca2+ pool that occurs during maturation, undermined the Ca2+ oscillatory ability of matured eggs and their activation potential. Collectively, this study supports a role for IP3R1 and C-IP3R1 in regulating Ca2+ homeostasis and the ER Ca2+ content during oocyte maturation. Nevertheless, the role of C-IP3R1 on Ca2+ homeostasis in aged eggs seems minor, as in MII eggs the majority of endogenous IP3R1 remains intact and C-IP3R1 undergoes rapid turnover. PMID:24692207

  15. The role of mounds in promoting water-exchange in the egg-tending burrows of monogamous goby, Valenciennea longipinnis (Lay et Bennett).

    PubMed

    Takegaki; Nakazono

    2000-10-25

    Valenciennea longipinnis spawns monogamously in a burrow. After spawning, the paired female constructs a conspicuous mound on the burrow by carrying and piling up substratum-derived materials while the male tends eggs in the burrow until hatching occurs. In this study, the mounds of V. longipinnis were tested in the field to confirm their function of promoting water-exchange in the burrow, and their ecological role was examined in relation to egg care by the male. The mound of V.longipinnis promoted water-exchange in the burrow, contributing to the provision of external oxygenated sea water into the burrow. Therefore, dissolved oxygen (DO) concentrations in the burrow with a mound were significantly higher than those without a mound. Although male egg-tending behaviour (e.g., fanning) may also promote water-exchange in the burrow, the water-exchange appeared to depend mainly on the hydrodynamic effect. Removals of the mound and paired female on the day of spawning led to high rates of egg-desertion by males. Since the frequency and time of fanning increase with a decrease of DO concentration in the burrow, the egg-desertion may result from an increased parental cost to males due to the decrease of water-exchange without a mound. This was supported by the fact that the DO concentrations on the day after mound removal were significantly lower in the egg-deserted burrows (measured before desertions) than in burrows not deserted by the male. Moreover, removals of paired females only also led to higher desertion rates. After removal of the female, the mound gradually collapsed by wave action and other factors, and the surface of the mound was covered with planktonic materials. Such a mound of poor quality may provide little water-exchange, which may lead to the egg-desertion by males. These results indicated that mound maintenance by females during the egg-tending period has an important role in the success of parental care by males.

  16. The potency of chicken egg yolk immunoglobulin (IgY) specific as immunotherapy to Mycobacterium tuberculosis infection

    PubMed Central

    Sudjarwo, Sri Agus; Eraiko, Koerniasari; Sudjarwo, Giftania Wardani; Koerniasari

    2017-01-01

    The aim of this study was to characterize of chicken egg yolk immunoglobulins (IgYs) specific as immunotherapy to Mycobacterium tuberculosis complex (MTBC) infection. Lohmann laying hens were immunized intramuscularly with antigenic of MTBC. Egg yolk was separated from egg white, and IgY antibody was then purified by multiple polyethylene glycols 6000 extraction and ammonium sulfate purification steps. The IgY anti-MTBC concentration in egg yolk increased at 2 weeks and it reached a maximum at 4 weeks after immunization. After 6 weeks, the levels of IgY anti-MTBC decreased gradually. The antibody of MTBC was detected and produces a specific line of precipitation in agar gel precipitation test beginning the week 2 after the first immunization. Analysis of results obtained with ELISA showed a significant increase in the MTBC specific antibodies after 2 weeks and reached a plateau at 4 weeks from the booster immunization. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed the IgY preparation to be pure and dissociated into protein bands with molecular weights of 112, 78, 69, 49, and 28 kDa and Western blot analysis shown the presence of anti-MTBC IgY in egg yolks, with molecular weights of approximately 78 kDa. These results suggested that egg yolk could be a practical strategy in large-scale production of specific anti-MTBC IgY for immunotherapy of TBC. PMID:28795022

  17. The potency of chicken egg yolk immunoglobulin (IgY) specific as immunotherapy to Mycobacterium tuberculosis infection.

    PubMed

    Sudjarwo, Sri Agus; Eraiko, Koerniasari; Sudjarwo, Giftania Wardani; Koerniasari

    2017-01-01

    The aim of this study was to characterize of chicken egg yolk immunoglobulins (IgYs) specific as immunotherapy to Mycobacterium tuberculosis complex (MTBC) infection. Lohmann laying hens were immunized intramuscularly with antigenic of MTBC. Egg yolk was separated from egg white, and IgY antibody was then purified by multiple polyethylene glycols 6000 extraction and ammonium sulfate purification steps. The IgY anti-MTBC concentration in egg yolk increased at 2 weeks and it reached a maximum at 4 weeks after immunization. After 6 weeks, the levels of IgY anti-MTBC decreased gradually. The antibody of MTBC was detected and produces a specific line of precipitation in agar gel precipitation test beginning the week 2 after the first immunization. Analysis of results obtained with ELISA showed a significant increase in the MTBC specific antibodies after 2 weeks and reached a plateau at 4 weeks from the booster immunization. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed the IgY preparation to be pure and dissociated into protein bands with molecular weights of 112, 78, 69, 49, and 28 kDa and Western blot analysis shown the presence of anti-MTBC IgY in egg yolks, with molecular weights of approximately 78 kDa. These results suggested that egg yolk could be a practical strategy in large-scale production of specific anti-MTBC IgY for immunotherapy of TBC.

  18. Determination of cortisol in lake sturgeon (Acipenser fulvescens) eggs by liquid chromatography tandem mass spectrometry.

    PubMed

    Bussy, Ugo; Wassink, Lydia; Scribner, Kim T; Li, Weiming

    2017-01-01

    Quantifying cortisol concentrations in fish eggs is important to understand the effects of environmental conditions on maternal physiological condition and on egg provisioning and quality. Data are particularly relevant to studies of the ecology of threatened species such as lake sturgeon (Aciperser fulvescens) as well as assessments of larval physical and behavioral phenotypes, fish health and caviar quality in sturgeon aquaculture. This study focuses on development of bioanalytical methods for high sensitivity and robust determination of cortisol in sturgeon eggs. Sample preparation was optimized after investigating protein precipitation and liquid-liquid extraction techniques. Ethyl acetate was found to be the most efficient solvent (recovery parameter) and also provided the best sample clean up (matrix effect parameter). The method was determined to be linear for cortisol concentrations between 0.025 and 100ng/mL. The limits of detection and quantification were 0.025 and 0.1ng/mL respectively. Intra- and inter-day performances of the method were validated at three concentrations (0.25; 10 and 100ng/mL). The method was applied to field-collected samples for the determination of endogenous cortisol in lake sturgeon eggs. Cortisol was detected in all egg samples and statistical analysis showed significant differences between fertilized and non-fertilized eggs. Copyright © 2016 Elsevier B.V. All rights reserved.

  19. Skin prick testing with extensively heated milk or egg products helps predict the outcome of an oral food challenge: a retrospective analysis.

    PubMed

    Faraj, Zein; Kim, Harold L

    2012-07-11

    Cow's milk and hen's egg are the most frequently encountered food allergens in the pediatric population. Skin prick testing (SPT) with commercial extracts followed by an oral food challenge (OFC) are routinely performed in the diagnostic investigation of these children. Recent evidence suggests that milk-allergic and/or egg-allergic individuals can often tolerate extensively heated (EH) forms of these foods. This study evaluated the predictive value of a negative SPT with EH milk or egg in determining whether a child would tolerate an OFC to the EH food product. Charts from a single allergy clinic were reviewed for any patient with a negative SPT to EH milk or egg, prepared in the form of a muffin. Data collected included age, sex, symptoms of food allergy, co-morbidities and the success of the OFC to the muffin. Fifty-eight patients had negative SPTs to the EH milk or egg in a muffin and underwent OFC to the appropriate EH food in the outpatient clinic. Fifty-five of these patients tolerated the OFC. The negative predictive value for the SPT with the EH food product was 94.8%. SPT with EH milk or egg products was predictive of a successful OFC to the same food. Larger prospective studies are required to substantiate these findings.

  20. Extreme intra-clutch egg size dimorphism is not coupled with corresponding differences in antioxidant capacity and stable isotopes between eggs.

    PubMed

    Poisbleau, Maud; Beaulieu, Michaël; Dehnhard, Nina; Demongin, Laurent; Lepoint, Gilles; Sturaro, Nicolas; Eens, Marcel

    2017-03-01

    Oviparous females need to allocate resources optimally to their eggs in order to maximize their fitness. Among these resources, dietary antioxidants, acquired by females and transferred to the eggs during egg formation, can greatly affect the development and survival of the embryo and chick. In crested penguins, incubation starts after the second and last egg is laid and, as opposed to many other bird species, this egg hatches first, thereby enhancing the survival of the chick. Here, we assessed whether antioxidant and isotopic composition could underlie these differences between eggs within clutches of southern rockhopper penguins (Eudyptes chrysocome chrysocome). The second-laid egg had higher total antioxidant capacity than the first-laid egg, although this was not due to higher antioxidant concentration but to its higher mass. This suggests that resources are allocated by females at a constant rate in both eggs within clutches. Accordingly, we found a strong correlation for isotopic compositions between eggs suggesting that resources were allocated similarly to each egg within the clutch. Overall, we found little evidence for a significant role of antioxidant and isotopic compositions to explain differences in terms of embryo/chick development between eggs in crested penguins. However, since our results suggest a constant rate of antioxidant transfer from females to eggs, limiting the mass of the first-laid egg might represent a strategy for females to spare antioxidant defences and preserve self-maintenance. Copyright © 2017 Elsevier Inc. All rights reserved.

  1. Optical properties of β-Ga2O3 nanorods synthesized by a simple and cost-effective method using egg white solution

    NASA Astrophysics Data System (ADS)

    Phumying, Santi; Labauyai, Sarawut; Chareonboon, Wirat; Phokha, Sumalin; Maensiri, Santi

    2015-06-01

    In this paper, we report on the optical properties of gallium oxide (β-Ga2O3) nanorods synthesized by a simple, cost-effective and environment-friendly method using gallium(III) nitrate hydrate and freshly extracted egg white (ovalbumin) in an aqueous medium. The extracted egg white acted as a matrix for the entrapment of gallium ions to generate a gelled precursor. The structure of the prepared samples was studied by X-ray diffraction and Raman spectroscopy to confirm the formation of β-Ga2O3 with a monoclinic structure after calcination of the precursor in air at 750, 850, and 950 °C for 3 h. Scanning electron microscopy images revealed the morphology and formation of nanorods with different sizes and shapes in the samples, resulting from the effect of the calcination temperature. All the samples showed a strong UV absorption with the band gap in the range of 3.87-3.97 eV. Room-temperature photoluminescence spectra of all the samples also showed a strong UV emission. The UV emission results were discussed based on the basis of charge recombination.

  2. Larvicidal & ovicidal efficacy of Pithecellobium dulce (Roxb.) Benth. (Fabaceae) against Anopheles stephensi Liston & Aedes aegypti Linn. (Diptera: Culicidae).

    PubMed

    Govindarajan, M; Rajeswary, M; Sivakumar, R

    2013-01-01

    In view of the recently increased interest in developing plant origin insecticides as an alternative to chemical insecticide, this study was undertaken to assess the larvicidal and ovicidal potential of the crude hexane, benzene, chloroform, ethyl acetate and methanol solvent extracts from the medicinal plant Pithecellobium dulce against the mosquito vectors, Anopheles stephensi and Aedes aegypti (Diptera: Culicidae). Larvicidal activity of P. dulce plant extracts was studied in the range of 60 to 450 mg/l against early third instar larvae of An. stephensi and Ae. aegypti in the laboratory. The larval mortality was observed after 24 h of exposure. The ovicidal activity was determined against An. stephensi and Ae. aegypti to various concentrations ranging from 100 to 750 mg/l under the laboratory conditions. Mean per cent hatchability of the eggs were observed after 48 h post treatment. All leaf and seed extracts showed moderate larvicidal and ovicidal effects; however, the highest larval mortality was found in methanol extract of leaf of P. dulce against the larvae of An. stephensi and Ae. aegypti with the LC₅₀ and LC₉₀ values 145.43, 155.78 mg/l and 251.23, 279.73 mg/l, respectively. The per cent hatchability was inversely proportional to the concentration of extract and directly proportional to the eggs. Zero hatchability was observed at 400 mg/l for leaf methanol extract and 625 mg/l for seed methanol extract of P. dulce against An. stephensi and Ae. aegypti, respectively. Compared to leaf extracts, seed extracts have low potency against the two mosquitoes. The present results suggest that the leaf and seed extracts of P. dulce have the potential to be used as an ideal eco-friendly approach for the control of mosquitoes.

  3. Egg and Egg-Derived Foods: Effects on Human Health and Use as Functional Foods

    PubMed Central

    Miranda, Jose M.; Anton, Xaquin; Redondo-Valbuena, Celia; Roca-Saavedra, Paula; Rodriguez, Jose A.; Lamas, Alexandre; Franco, Carlos M.; Cepeda, Alberto

    2015-01-01

    Eggs are sources of protein, fats and micronutrients that play an important role in basic nutrition. However, eggs are traditionally associated with adverse factors in human health, mainly due to their cholesterol content. Nowadays, however, it is known that the response of cholesterol in human serum levels to dietary cholesterol consumption depends on several factors, such as ethnicity, genetic makeup, hormonal factors and the nutritional status of the consumer. Additionally, in recent decades, there has been an increasing demand for functional foods, which is expected to continue to increase in the future, owing to their capacity to decrease the risks of some diseases and socio-demographic factors such as the increase in life expectancy. This work offers a brief overview of the advantages and disadvantages of egg consumption and the potential market of functional eggs, and it explores the possibilities of the development of functional eggs by technological methods. PMID:25608941

  4. Survival, physical and physiological changes of Taenia hydatigena eggs under different conditions of water stress.

    PubMed

    Sánchez Thevenet, Paula; Alvarez, Hector Manuel; Basualdo, Juan Angel

    2017-06-01

    Taenia hydatigena eggs were investigated for morphological and physiological changes under water stress conditions. Fresh eggs were exposed at 31%, 47% and 89% of relative humidity (RH), and survival, size and ultrastructural changes were accounted up to 365 days of exposition. The article shows how each RH environment affects the vitality of the eggs. Results of this study suggest that T. hydatigena eggs have mechanisms to withstand water stress, indicating that the eggs clustering improves protection against desiccation, and that endogenous metabolism using triacylglycerols play an important role in the maintenance of embryo vitality under low, medium and high relative humidity conditions. This contributes to understanding the water stress resistance mechanism in eggs belonging to Taeniidae family. The findings shown herein have provided a basis to better comprehend basic biology and epidemiology of the cysticercosis caused by T. hydatigena. Copyright © 2017 Elsevier Inc. All rights reserved.

  5. The role of plasmalogen in the oxidative stability of neutral lipids and phospholipids.

    PubMed

    Wang, Guang; Wang, Tong

    2010-02-24

    The role of ethanolamine plasmalogen extracted from bovine brain (BBEP) in maintaining oxidative stability of bulk soybean oil and liposome made with egg phospholipids (PL) was studied. In a purified soybean oil (PSO), the addition of 200 and 1000 ppm of BBEP promoted lipid oxidation at rates of 0.037 and 0.071 (all rates in ln (PV) h(-1), and PV stands for peroxide value), whereas soy lecithin (SL) added in the same amount showed a trend similar to the PSO blank, which had an oxidation rate of 0.025. The PSO with BBEP was susceptible to cupric ion catalyzed oxidation, in that the oil was oxidized much more quickly than the PSO with SL and cupric ion. In commercial soybean oil (CSO) with the presence of tocopherols, SL at 1000 ppm acted synergistically as an antioxidant with the natural tocopherols, but addition of BBEP accelerated lipid oxidation, as evidenced by the oxidative stability index (OSI) test. In the egg PL liposome, the BBEP caused a fast breakdown of the lipid hydroperoxides and consequently promoted more thiobarbituric acid reactive substance (TBARS) formation. The PL oxidation in the presence of copper in the liposome was not affected by the BBEP, which indicates that the hypothesis of ethanolamine plasmalogen (EthPm) chelating cupric ion as the antioxidation mechanism was not supported. The addition of cumene hydroperoxide to the egg PL liposome promoted lipid oxidation, as indicated by a fast development of PV and TBARS. However, the result with cumene hydroperoxide failed to differentiate the effect of BBEP and SL and their concentration on lipid oxidation. On the basis of the observations from this study, we conclude that EthPm is not an antioxidant but rather a pro-oxidant in a bulk lipid system, and it has no significant antioxidant effect for PL oxidation in the liposome.

  6. Localization of the Kinesin-like Protein Xklp2 to Spindle Poles Requires a Leucine Zipper, a Microtubule-associated Protein, and Dynein

    PubMed Central

    Wittmann, Torsten; Boleti, Haralabia; Antony, Claude; Karsenti, Eric; Vernos, Isabelle

    1998-01-01

    Xklp2 is a plus end–directed Xenopus kinesin-like protein localized at spindle poles and required for centrosome separation during spindle assembly in Xenopus egg extracts. A glutathione-S-transferase fusion protein containing the COOH-terminal domain of Xklp2 (GST-Xklp2-Tail) was previously found to localize to spindle poles (Boleti, H., E. Karsenti, and I. Vernos. 1996. Cell. 84:49–59). Now, we have examined the mechanism of localization of GST-Xklp2-Tail. Immunofluorescence and electron microscopy showed that Xklp2 and GST-Xklp2-Tail localize specifically to the minus ends of spindle pole and aster microtubules in mitotic, but not in interphase, Xenopus egg extracts. We found that dimerization and a COOH-terminal leucine zipper are required for this localization: a single point mutation in the leucine zipper prevented targeting. The mechanism of localization is complex and two additional factors in mitotic egg extracts are required for the targeting of GST-Xklp2-Tail to microtubule minus ends: (a) a novel 100-kD microtubule-associated protein that we named TPX2 (Targeting protein for Xklp2) that mediates the binding of GST-Xklp2-Tail to microtubules and (b) the dynein–dynactin complex that is required for the accumulation of GST-Xklp2-Tail at microtubule minus ends. We propose two molecular mechanisms that could account for the localization of Xklp2 to microtubule minus ends. PMID:9813089

  7. β1 Integrin is an Adhesion Protein for Sperm Binding to Eggs

    PubMed Central

    Baessler, Keith A.; Lee, Younjoo; Sampson, Nicole S.

    2009-01-01

    We investigated the role of β1 integrin in mammalian fertilization and the mode of inhibition of fertilinβ-derived polymers. We determined that polymers displaying the Glu-Cys-Asp peptide from the fertilinβ disintegrin domain mediate inhibition of mammalian fertilization through a β1 integrin receptor on the egg surface. Inhibition of fertilization is a consequence of competition with sperm binding to the cell surface, not activation of an egg-signaling pathway. The presence of the β1 integrin on the egg surface increases the rate of sperm attachment, but does not alter the total number of sperm that can attach or fuse to the egg. We conclude that the presence of β1 integrin enhances the initial adhesion of sperm to the egg plasma membrane and that subsequent attachment and fusion are mediated by additional egg and sperm proteins present in the β1 integrin complex. Therefore, the mechanisms by which sperm fertilize wild-type and β1 knockout eggs are different. PMID:19338281

  8. Mechanosensitive neurons on the internal reproductive tract contribute to egg-laying-induced acetic acid attraction in Drosophila

    PubMed Central

    Gou, Bin; Liu, Ying; Guntur, Ananya R.; Stern, Ulrich; Yang, Chung-Hui

    2014-01-01

    Selecting a suitable site to deposit their eggs is an important reproductive need of Drosophila females. While their choosiness towards egg-laying sites is well documented, the specific neural mechanism that activates females’ search for attractive egg-laying sites is not known. Here we show that distention/contraction of females’ internal reproductive tract triggered by egg-delivery through the tract plays a critical role in activating such search. We found that females start to exhibit acetic acid attraction prior to depositing each egg but no attraction when they are not laying eggs. Artificially distending the reproductive tract triggers acetic acid attraction in non-egg-laying females whereas silencing the mechanosensitive neurons we identified that can sense the contractile status of the tract eliminates such attraction. Our work uncovers the circuit basis of an important reproductive need of Drosophila females and provides a simple model to dissect the neural mechanism that underlies a reproductive need-induced behavioral modification. PMID:25373900

  9. Comparative effects of Cymbopogon schoenanthus essential oil and piperitone on Callosobruchus maculatus development.

    PubMed

    Ketoh, Guillaume K; Koumaglo, Honore K; Glitho, Isabelle A; Huignard, Jacques

    2006-12-01

    The insecticidal activity of crude essential oil extracted from Cymbopogon schoenanthus and of its main constituent, piperitone, was assessed on different developmental stages of Callosobruchus maculatus. Piperitone was more toxic to adults with a LC(50) value of 1.6 microl/l vs. 2.7 microl/l obtained with the crude extract. Piperitone inhibited the development of newly laid eggs and of neonate larvae, but was less toxic than the crude extract to individuals developing inside the seeds.

  10. Comparison of environmental and egg microbiology associated with conventional and free-range laying hen management.

    PubMed

    Jones, D R; Anderson, K E; Musgrove, M T

    2011-09-01

    Eggs from alternative production practices are a growing niche in the market. Meeting consumer requests for greater diversity in retail egg options has resulted in some unique challenges such as understanding the food safety implications of eggs from alternative production practices. A study was conducted to determine what, if any, differences exist between nest run conventional cage-produced eggs and free range-produced eggs. A sister flock of brown egg layers was maintained in conventional cage and free-range production with egg and environmental sampling every 6 wk from 20 to 79 wk of age. Aerobic, coliform, and yeast and mold populations were monitored. Environmental microbial levels were not always indicative of egg contamination levels. When significant differences (P < 0.05 and P < 0.0001, dependent on season) were observed among treatments for coliforms, shell contamination levels of free-range nest box eggs and free-range floor eggs were always greater than those of conventional cage eggs, which remained low throughout the study (0.42-0.02 log cfu/mL). Shell yeast and mold levels were significantly greater in free-range floor eggs than in free-range nest box eggs and conventional cage eggs throughout the entire study. Egg contents contamination levels were extremely low for all monitored populations and treatments. Season of the year played a role in both environmental and egg microbial levels. Winter had the lowest levels of all populations monitored for all treatments, except for aerobic free-range floor egg shell emulsions, which were increased (3.6 log cfu/mL). Understanding the differences in microbial populations present on conventional cage-produced and free range-produced eggs can lead to the development of effective cleaning procedures, enhancing food safety.

  11. Quantification of antibody (IgY) titers in hen eggs following immunization and their use in detecting cell surface molecules on nitrocellulose membranes.

    PubMed

    Ruan, Guang-Ping; Ma, Li; Meng, Xiao-Jing; Meng, Min-Jie; Wang, Xiao-Ning; Lin, Ying; Wu, Zheng-Qiang; He, Xiao-Wei; Wang, Ju-Fang; Zhu, Yong

    2007-01-01

    HLA-A*0201 alpha chain and beta2m were expressed from a prokaryotic system, and after refolding and purification, the alpha chain and beta2m were used to immunize eight laying hens. The titer of egg yolk antibody against alpha chain increased from 10(2) to 10(5.3) The titer of egg yolk antibody against beta2m increased from 10(1) to 10(4.7). The extent of titer increase is similar between the two antigens. An average of 135 mg purified polyclonal antibody (IgY) can be easily obtained from one egg yolk. The use of egg collection rather than serum collection is compatible with modern animal protection regulations. An average of 28 eggs were obtained from a laying hen every month, with a total amount of 3780 mg immunoglobulin extracted from one immunized hen every month, which would be equivalent to 630 mL of serum or 1260 mL of blood per month. Chickens are an optimal host for the production of polyclonal antibodies with high titer and high yield. Purified IgY was labeled with horseradish peroxidase and reacted with PBMC on nitrocellulose membranes indicating that the antibody can bind to the native conformation of class I HLA molecule on PBMC.

  12. Advances in ultra-high performance liquid chromatography coupled to tandem mass spectrometry for sensitive detection of several food allergens in complex and processed foodstuffs.

    PubMed

    Planque, M; Arnould, T; Dieu, M; Delahaut, P; Renard, P; Gillard, N

    2016-09-16

    Sensitive detection of food allergens is affected by food processing and foodstuff complexity. It is therefore a challenge to detect cross-contamination in food production that could endanger an allergic customer's life. Here we used ultra-high performance liquid chromatography coupled to tandem mass spectrometry for simultaneous detection of traces of milk (casein, whey protein), egg (yolk, white), soybean, and peanut allergens in different complex and/or heat-processed foodstuffs. The method is based on a single protocol (extraction, trypsin digestion, and purification) applicable to the different tested foodstuffs: chocolate, ice cream, tomato sauce, and processed cookies. The determined limits of quantitation, expressed in total milk, egg, peanut, or soy proteins (and not soluble proteins) per kilogram of food, are: 0.5mg/kg for milk (detection of caseins), 5mg/kg for milk (detection of whey), 2.5mg/kg for peanut, 5mg/kg for soy, 3.4mg/kg for egg (detection of egg white), and 30.8mg/kg for egg (detection of egg yolk). The main advantage is the ability of the method to detect four major food allergens simultaneously in processed and complex matrices with very high sensitivity and specificity. Copyright © 2016 Elsevier B.V. All rights reserved.

  13. Differential label-free quantitative proteomic analysis of avian eggshell matrix and uterine fluid proteins associated with eggshell mechanical property.

    PubMed

    Sun, Congjiao; Xu, Guiyun; Yang, Ning

    2013-12-01

    Eggshell strength is a crucial economic trait for table egg production. During the process of eggshell formation, uncalcified eggs are bathed in uterine fluid that plays regulatory roles in eggshell calcification. In this study, a label-free MS-based protein quantification technology was used to detect differences in protein abundance between eggshell matrix from strong and weak eggs (shell matrix protein from strong eggshells and shell matrix protein from weak eggshells) and between the corresponding uterine fluids bathing strong and weak eggs (uterine fluid bathing strong eggs and uterine fluid bathing weak eggs) in a chicken population. Here, we reported the first global proteomic analysis of uterine fluid. A total of 577 and 466 proteins were identified in uterine fluid and eggshell matrix, respectively. Of 447 identified proteins in uterine fluid bathing strong eggs, up to 357 (80%) proteins were in common with proteins in uterine fluid bathing weak eggs. Similarly, up to 83% (328/396) of the proteins in shell matrix protein from strong eggshells were in common with the proteins in shell matrix protein from weak eggshells. The large amount of common proteins indicated that the difference in protein abundance should play essential roles in influencing eggshell strength. Ultimately, 15 proteins mainly relating to eggshell matrix specific proteins, calcium binding and transportation, protein folding and sorting, bone development or diseases, and thyroid hormone activity were considered to have closer association with the formation of strong eggshell. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  14. Mechanistic basis of adaptive maternal effects: egg jelly water balance mediates embryonic adaptation to acidity in Rana arvalis.

    PubMed

    Shu, Longfei; Suter, Marc J-F; Laurila, Anssi; Räsänen, Katja

    2015-11-01

    Environmental stress, such as acidification, can challenge persistence of natural populations and act as a powerful evolutionary force at ecological time scales. The ecological and evolutionary responses of natural populations to environmental stress at early life-stages are often mediated via maternal effects. During early life-stages, maternal effects commonly arise from egg coats (the extracellular structures surrounding the embryo), but the role of egg coats has rarely been studied in the context of adaptation to environmental stress. Previous studies on the moor frog Rana arvalis found that the egg coat mediated adaptive divergence along an acidification gradient in embryonic acid stress tolerance. However, the exact mechanisms underlying these adaptive maternal effects remain unknown. Here, we investigated the role of water balance and charge state (zeta potential) of egg jelly coats in embryonic adaptation to acid stress in three populations of R. arvalis. We found that acidic pH causes severe water loss in the egg jelly coat, but that jelly coats from an acid-adapted population retained more water than jelly coats from populations not adapted to acidity. Moreover, embryonic acid tolerance (survival at pH 4.0) correlated with both water loss and charge state of the jelly, indicating that negatively charged glycans influence jelly water balance and contribute to embryonic adaptation to acidity. These results indicate that egg coats can harbor extensive intra-specific variation, probably facilitated in part via strong selection on water balance and glycosylation status of egg jelly coats. These findings shed light on the molecular mechanisms of environmental stress tolerance and adaptive maternal effects.

  15. Roles of calcium and pH in activation of eggs of the medaka fish, Oryzias latipes

    PubMed Central

    1983-01-01

    Unfertilized eggs of the medaka fish (Oryzias latipes) were injected with pH-buffered calcium buffers. Medaka egg activation is accompanied by a transient increase in cytoplasmic free calcium (Gilkey, J. C., L. F. Jaffe, E. B. Ridgway, and G. T. Reynolds, 1978, J. Cell Biol., 76:448-466). The calcium buffer injections demonstrated that (a) the threshold free calcium required to elicit the calcium transient and activate the egg is between 1.7 and 5.1 microM at pH 7.0, well below the 30 microM reached during the transient, and (b) buffers which hold free calcium below threshold prevent activation of the buffered region in subsequently fertilized eggs. Therefore an increase in free calcium is necessary and sufficient to elicit the calcium transient, and the calcium transient is necessary to activate the egg. Further, these results are additional proof that the calcium transient is initiated and propagated through the cytoplasm by a mechanism of calcium- stimulated calcium release. Finally, a normal calcium transient must propagate through the entire cytoplasm to ensure normal development. Unfertilized eggs were injected with pH buffers to produce short-term, localized changes in cytoplasmic pH. The eggs were then fertilized at various times after injection. In other experiments, unfertilized and fertilized eggs were exposed to media containing either NH4Cl or CO2 to produce longer term, global changes in cytoplasmic pH. These treatments neither activated the eggs nor interfered with the normal development of fertilized eggs, suggesting that even if a natural change in cytoplasmic pH is induced by activation, it has no role in medaka egg development. The injected pH buffers altered the rate of propagation of the calcium transient through the cytoplasm, suggesting that the threshold free calcium required to trigger calcium-stimulated calcium release might be pH dependent. The results of injection of pH-buffered calcium buffers support this conjecture: for a tenfold increase in hydrogen ion concentration, free calcium must also be raised tenfold to elicit the calcium transient. PMID:6411737

  16. Intra-Amniotic Administration (Gallus gallus) of Cicer arietinum and Lens culinaris Prebiotics Extracts and Duck Egg White Peptides Affects Calcium Status and Intestinal Functionality

    PubMed Central

    Hou, Tao; Glahn, Raymond P.; Tako, Elad

    2017-01-01

    Calcium (Ca) is one of the most abundant inorganic elements in the human body and has many important physiological roles. Prebiotics and bioactive peptides are two important substances used to promote calcium uptake. However, the difference in mechanisms of the calcium uptake from these two supplements is not clear. By using the Gallus gallus model and the intra-amniotic administration procedure, the aim of this study was to investigate whether Ca status, intestinal functionality, and health-promoting bacterial populations were affected by prebiotics extracted from chickpea and lentil, and duck egg white peptides (DPs). Eleven groups (non-injected; 18 MΩ H2O; 4 mmol/L CaCl2; 50 mg/mL chickpea + 4 mmol/L CaCl2; 50 mg/mL lentil + 4 mmol/L CaCl2; 40 mg/mL DPs + 4 mmol/L CaCl2; 5 mg/mL Val-Ser-Glu-Glu (VSEE) + 4 mmol/L CaCl2; 50 mg/mL chickpea; 50 mg/mL lentil; 40 mg/mL DPs; 5 mg/mL VSEE) were utilized. Upon hatch, blood, cecum, small intestine, liver and bone were collected for assessment of serum bone alkaline phosphate level (BALP), the relative abundance of intestinal microflora, expression of Ca-related genes, brush border membrane (BBM) functional genes, and liver and bone mineral levels, respectively. The BALP level increased in the presence of lentil, DPs and VSEE (p < 0.05). The relative abundance of probiotics increased significantly (p < 0.05) by VSEE + Ca and chickpea. The expression of CalbindinD9k (Ca transporter) increased (p < 0.05) in Ca, chickpea + Ca and lentil + Ca groups. In addition, the brush border membrane functionality genes expressions increased (p < 0.05) by the chickpea or lentil extracts. Prebiotics and DPs beneficially affected the intestinal microflora and duodenal villus surface area. This research expands the understanding of the prebiotics’ properties of chickpea and lentil extracts, and peptides’ effects on calcium metabolism and gut health. PMID:28754012

  17. Intra-Amniotic Administration (Gallus gallus) of Cicer arietinum and Lens culinaris Prebiotics Extracts and Duck Egg White Peptides Affects Calcium Status and Intestinal Functionality.

    PubMed

    Hou, Tao; Kolba, Nikolai; Glahn, Raymond P; Tako, Elad

    2017-07-21

    Calcium (Ca) is one of the most abundant inorganic elements in the human body and has many important physiological roles. Prebiotics and bioactive peptides are two important substances used to promote calcium uptake. However, the difference in mechanisms of the calcium uptake from these two supplements is not clear. By using the Gallus gallus model and the intra-amniotic administration procedure, the aim of this study was to investigate whether Ca status, intestinal functionality, and health-promoting bacterial populations were affected by prebiotics extracted from chickpea and lentil, and duck egg white peptides (DPs). Eleven groups (non-injected; 18 MΩ H₂O; 4 mmol/L CaCl₂; 50 mg/mL chickpea + 4 mmol/L CaCl₂; 50 mg/mL lentil + 4 mmol/L CaCl₂; 40 mg/mL DPs + 4 mmol/L CaCl₂; 5 mg/mL Val-Ser-Glu-Glu (VSEE) + 4 mmol/L CaCl₂; 50 mg/mL chickpea; 50 mg/mL lentil; 40 mg/mL DPs; 5 mg/mL VSEE) were utilized. Upon hatch, blood, cecum, small intestine, liver and bone were collected for assessment of serum bone alkaline phosphate level (BALP), the relative abundance of intestinal microflora, expression of Ca-related genes, brush border membrane (BBM) functional genes, and liver and bone mineral levels, respectively. The BALP level increased in the presence of lentil, DPs and VSEE ( p < 0.05). The relative abundance of probiotics increased significantly ( p < 0.05) by VSEE + Ca and chickpea. The expression of CalbindinD9k (Ca transporter) increased ( p < 0.05) in Ca, chickpea + Ca and lentil + Ca groups. In addition, the brush border membrane functionality genes expressions increased ( p < 0.05) by the chickpea or lentil extracts. Prebiotics and DPs beneficially affected the intestinal microflora and duodenal villus surface area. This research expands the understanding of the prebiotics' properties of chickpea and lentil extracts, and peptides' effects on calcium metabolism and gut health.

  18. Oviposition Deterrent Activities of Pachyrhizus erosus Seed Extract and Other Natural Products on Plutella xylostella (Lepidoptera: Plutellidae)

    PubMed Central

    Basukriadi, Adi; Wilkins, Richard M.

    2014-01-01

    Abstract An extract of a rotenone-containing plant yam bean, Pachyrhizus erosus (L.) Urban, seeds was tested against the diamondback moth, Plutella xylostella (L.) in a greenhouse to determine its potential as an oviposition deterrent and compared with coumarin and rutin, known as diamondback moth oviposition deterrent compounds, rotenone, and an extract of Peruvian cube root, at a concentration of 0.5% (w/v). Oviposition deterrent index (ODI) was used to determine effects of extracts or compounds in inhibiting oviposition of diamondback moth. Coumarin showed a stronger deterrent effect than the yam bean seed extract with a higher ODI value. On the contrary, rotenone, rutin, and the cube root extract, containing 6.7% (w/w) of rotenone, showed no significant deterrent effects having low or negative ODI values, suggesting that the deterrent effect of the yam bean seed extract is not due to rotenone content of the yam bean seeds. The extract of yam bean seed and coumarin partially deterred the moth from laying eggs on treated leaves in a concentration-dependent manner. The effective concentration for 50% deterrency of coumarin and the yam bean seed extract were 0.11 and 0.83% (w/v), respectively. However, the yam bean seed extract showed a residual deterrent effect on the moth even at 3 d after the treatment and is probably because of its low volatile nature. A long-term deterrency of the yam bean seed extract is an advantage over coumarins. Both the yam bean seed extract and coumarin deterred diamondback moth from laying eggs in total darkness, indicating their nonvisual deterrent effect. This made the extract an effective deterrence to diamondback moth in light and in darkness. To conclude, this study revealed the potential of the crude extract of the yam bean seed to prevent diamondback moth from ovipositing on its plant host. PMID:25525107

  19. Oviposition deterrent activities of Pachyrhizus erosus seed extract and other natural products on Plutella xylostella (Lepidoptera: Plutellidae).

    PubMed

    Basukriadi, Adi; Wilkins, Richard M

    2014-01-01

    An extract of a rotenone-containing plant yam bean, Pachyrhizus erosus (L.) Urban, seeds was tested against the diamondback moth, Plutella xylostella (L.) in a greenhouse to determine its potential as an oviposition deterrent and compared with coumarin and rutin, known as diamondback moth oviposition deterrent compounds, rotenone, and an extract of Peruvian cube root, at a concentration of 0.5% (w/v). Oviposition deterrent index (ODI) was used to determine effects of extracts or compounds in inhibiting oviposition of diamondback moth. Coumarin showed a stronger deterrent effect than the yam bean seed extract with a higher ODI value. On the contrary, rotenone, rutin, and the cube root extract, containing 6.7% (w/w) of rotenone, showed no significant deterrent effects having low or negative ODI values, suggesting that the deterrent effect of the yam bean seed extract is not due to rotenone content of the yam bean seeds. The extract of yam bean seed and coumarin partially deterred the moth from laying eggs on treated leaves in a concentration-dependent manner. The effective concentration for 50% deterrency of coumarin and the yam bean seed extract were 0.11 and 0.83% (w/v), respectively. However, the yam bean seed extract showed a residual deterrent effect on the moth even at 3 d after the treatment and is probably because of its low volatile nature. A long-term deterrency of the yam bean seed extract is an advantage over coumarins. Both the yam bean seed extract and coumarin deterred diamondback moth from laying eggs in total darkness, indicating their nonvisual deterrent effect. This made the extract an effective deterrence to diamondback moth in light and in darkness. To conclude, this study revealed the potential of the crude extract of the yam bean seed to prevent diamondback moth from ovipositing on its plant host. © The Author 2014. Published by Oxford University Press on behalf of the Entomological Society of America.

  20. Cysteine-rich secretory proteins (CRISP) and their role in mammalian fertilization.

    PubMed

    Cohen, Débora J; Maldera, Julieta A; Weigel Muñoz, Mariana; Ernesto, Juan I; Vasen, Gustavo; Cuasnicu, Patricia S

    2011-01-01

    Epididymal protein CRISPI is a member of the CRISP (Cysteine-RIch Secretory proteins) family and is involved in sperm-egg fusion through its interaction with complementary sites on the egg surface. Results from our laboratory have shown that this binding ability resides in a 12-amino-acid region corresponding to a highly conserved motif of the CRISP family, named Signature 2 (S2). In addition to this, our results revealed that CRISP1 could also be involved in the previous step of sperm binding to the zona pellucida, identifying a novel role for this protein in fertilization. As another approach to elucidate the participation of CRISP1 in fertilization, a mouse line containing a targeted disruption of CRISP1 was generated. Although CRISP1-deficient mice exhibited normal fertility, CRISP1-defficient sperm presented a decreased level of protein tyrosine phosphorylation during capacitation, and an impaired ability to fertilize both zona-intact and zona-free eggs in vitro, confirming the proposed roles for the protein in fertilization. Evidence obtained in our laboratory indicated that testicular CRISP2 would also be involved in sperm-egg fusion. Competition assays between CRISP1 and CRISP2, as well as the comparison of their corresponding S2 regions, suggest that both proteins bind to common complementary sites in the egg. Together, these results suggest a functional cooperation between CRISP1 and CRISP2 to ensure the success of fertilization.

  1. [Application of the elliptic fourier functions to the description of avian egg shape].

    PubMed

    Ávila, Dennis Denis

    2014-12-01

    Egg shape is difficult to quantify due to the lack of an exact formula to describe its geometry. Here I described a simple algorithm to characterize and compare egg shapes using Fourier functions. These functions can delineate any closed contour and had been previously applied to describe several biological objects. I described, step by step, the process of data acquisition, processing and the use of the SHAPE software to extract function coefficients in a study case. I compared egg shapes in three birds' species representing different reproductive strategies: Cuban Parakeet (Aratinga euops), Royal Tern (Thalasseus maximus) and Cuban Blackbird (Dives atroviolaceus). Using 73 digital pictures of eggs kept in Cuban scientific collections, I calculated Fourier descriptors with 4, 6, 8, 16 and 20 harmonics. Descriptors were reduced by a Principal Component Analysis and the scores of the eigen-values that account for 90% of variance were used in a Lineal Discriminant Function to analyze the possibility to differentiate eggs according to its shapes. Using four harmonics, the first five component accounted for 97% of shape variances; more harmonics diluted the variance increasing to eight the number of components needed to explain most of the variation. Convex polygons in the discriminant space showed a clear separation between species, allowing trustful discrimination (classification errors between 7-15%). Misclassifications were related to specific egg shape variability between species. In the study case, A. euops eggs were perfectly classified, but for the other species, errors ranged from 5 to 29% of misclassifications, in relation to the numbers or harmonics and components used. The proposed algorithm, despite its apparent mathematical complexity, showed many advantages to describe eggs shape allowing a deeper understanding of factors related to this variable.

  2. Histone H3K9 Trimethylase Eggless Controls Germline Stem Cell Maintenance and Differentiation

    PubMed Central

    Zhou, Jian; McDowell, William; Park, Jungeun; Haug, Jeff; Staehling, Karen; Tang, Hong; Xie, Ting

    2011-01-01

    Epigenetic regulation plays critical roles in the regulation of cell proliferation, fate determination, and survival. It has been shown to control self-renewal and lineage differentiation of embryonic stem cells. However, epigenetic regulation of adult stem cell function remains poorly defined. Drosophila ovarian germline stem cells (GSCs) are a productive adult stem cell system for revealing regulatory mechanisms controlling self-renewal and differentiation. In this study, we show that Eggless (Egg), a H3K9 methyltransferase in Drosophila, is required in GSCs for controlling self-renewal and in escort cells for regulating germ cell differentiation. egg mutant ovaries primarily exhibit germ cell differentiation defects in young females and gradually lose GSCs with time, indicating that Egg regulates both germ cell maintenance and differentiation. Marked mutant egg GSCs lack expression of trimethylated H3K9 (H3k9me3) and are rapidly lost from the niche, but their mutant progeny can still differentiate into 16-cell cysts, indicating that Egg is required intrinsically to control GSC self-renewal but not differentiation. Interestingly, BMP-mediated transcriptional repression of differentiation factor bam in marked egg mutant GSCs remains normal, indicating that Egg is dispensable for BMP signaling in GSCs. Normally, Bam and Bgcn interact with each other to promote GSC differentiation. Interestingly, marked double mutant egg bgcn GSCs are still lost, but their progeny are able to differentiate into 16-cell cysts though bgcn mutant GSCs normally do not differentiate, indicating that Egg intrinsically controls GSC self-renewal through repressing a Bam/Bgcn-independent pathway. Surprisingly, RNAi-mediated egg knockdown in escort cells leads to their gradual loss and a germ cell differentiation defect. The germ cell differentiation defect is at least in part attributed to an increase in BMP signaling in the germ cell differentiation niche. Therefore, this study has revealed the essential roles of histone H3K9 trimethylation in controlling stem cell maintenance and differentiation through distinct mechanisms. PMID:22216012

  3. Brominated flame retardants (BFRs) in eggs from birds of prey from Southern Germany, 2014.

    PubMed

    Vetter, Walter; Gallistl, Christoph; Schlienz, Annika; Preston, Theresa; Müller, Jens; von der Trenck, K Theo

    2017-12-01

    In Southern Germany, peregrine falcons (Falco peregrinus), which almost exclusively prey on other birds, are top predators of the terrestrial food chain. These animals accumulate persistent organic pollutants (POPs) and halogenated flame retardants (HFRs) with mothers transferring these lipophilic contaminants to their eggs. Here we analyzed unhatched eggs of eleven peregrine falcons and six of other species, and report concentrations of polybrominated diphenyl ethers (PBDEs), hexabromocyclododecane (HBCD), hexabromobenzene (HBB), 2,3-dibromopropyl-2,4,6-tribromophenyl ether (DPTE) and its metabolites, pentabromoethylbenzene (PBEB), pentabromotoluene (PBT), and tribromophenol (TBP). The extract of one purified peregrine falcon egg sample was comprehensively analyzed in a non-target (NT) approach by gas chromatography with mass spectrometry in the electron capture negative ion mode. A total of ∼400 polyhalogenated compounds were detected, among them dechloranes and possibly transformation products, two tetrabrominated metabolites of PBT and several compounds unknown to us which could not be identified. Copyright © 2017 Elsevier Ltd. All rights reserved.

  4. Nest sanitation elicits egg discrimination in cuckoo hosts.

    PubMed

    Yang, Canchao; Chen, Min; Wang, Longwu; Liang, Wei; Møller, Anders Pape

    2015-11-01

    Nest sanitation is a nearly universal behavior in birds, while egg discrimination is a more specific adaptation that has evolved to counter brood parasitism. These two behaviors are closely related with nest sanitation being the ancestral behavior, and it has been hypothesized to constitute a preadaptation for egg discrimination. However, previous studies found little evidence to support this hypothesis. Here, we conducted an empirical test of the association between nest sanitation and egg discrimination in the barn swallow (Hirundo rustica) by inserting a single non-mimetic model egg or a non-mimetic model egg plus half a peanut shell into host nests. Compared to the rejection rate of single model eggs, barn swallows significantly increased egg rejection frequency if a half peanut shell was simultaneously introduced. Our result for the first time shows the impact of nest sanitation on egg discrimination and demonstrates that nest sanitation can elicit egg discrimination in hosts of brood parasites. This study provided evidence for nest sanitation being a preadaptation to egg discrimination by facilitating egg rejection, thereby significantly advancing our understanding of avian cognition of foreign objects. Furthermore, we suggest that egg discrimination behavior in many accepters and intermediate rejecters may be lost or diluted. Such egg discrimination can be elicited and restored after nest sanitation, implying a sensitive and rapid phenotypic response to increased risk of parasitism. Our study offers a novel perspective for investigating the role of so-called intermediate rejecter individuals or species in the long-term coevolutionary cycle between brood parasites and their hosts.

  5. [Determination of canthaxanthin and astaxanthin in egg yolks by reversed phase high performance liquid chromatography with diode array detection].

    PubMed

    He, Kang-Hao; Zou, Xiao-Li; Liu, Xiang; Zeng, Hong-Yan

    2012-01-01

    A method using reversed phase high performance liquid chromatography (RP-HPLC) coupled with diode array detector (DAD) was developed for the simultaneous determination of canthaxanthin and astaxanthin in egg yolks. Samples were extracted with acetonitrile in ultrasonic bath for 20 minutes and then purified by freezing-lipid filtration and solid phase extraction (SPE). After being vaporized to dryness by nitrogen blowing and made up to volume with methanol, the extract solution was chromatographically separated in C18 column with a unitary mobile phase consisting of acetonitrile. The proposed method was validated in terms of linearity, precision, accuracy, and limit of detection (LOD). Regression analysis revealed a good linearity between peak area of each analyte and its concentration (r > or = 0.998). The intra- and inter-day relative standard deviations (RSDs) were less than 3.6% and 5.2%, respectively. LODs of canthaxanthin and astaxanthin were 0.035 and 0.027 microg/mL (S/N = 3). The average recoveries of canthaxanthin and astaxanthin were 91.5% and 88.7%. The proposed method is simple, fast and easy to apply.

  6. Effects of neem seed derivatives on behavioral and physiological responses of the Cosmopolites sordidus (Coleoptera: Curculionidae).

    PubMed

    Musabyimana, T; Saxena, R C; Kairu, E W; Ogol, C P; Khan, Z R

    2001-04-01

    Both in a choice and multi-choice laboratory tests, fewer adults of the banana root borer, Cosmopolites sordidus (Germar), settled under the corms of the susceptible banana "Nakyetengu" treated with 5% aqueous extract of neem seed powder or cake or 2.5 and 5% emulsified neem oil than on water-treated corms. Feeding damage by larvae on banana pseudostem discs treated with 5% extract of powdered neem seed, kernel, or cake, or 5% emulsified neem oil was significantly less than on untreated discs. The larvae took much longer to locate feeding sites, initiate feeding and bore into pseudostem discs treated with extract of powdered neem seed or kernel. Few larvae survived when confined for 14 d on neem-treated banana pseudostems; the survivors weighed two to four times less than the larvae developing on untreated pseudostems. Females deposited up to 75% fewer eggs on neem-treated corms. In addition, egg hatching was reduced on neem-treated corms. The higher the concentration of neem materials the more severe the effect.

  7. Participation of cysteine-rich secretory proteins (CRISP) in mammalian sperm-egg interaction.

    PubMed

    Cohen, Débora J; Busso, Dolores; Da Ros, Vanina; Ellerman, Diego A; Maldera, Julieta A; Goldweic, Nadia; Cuasnicu, Patricia S

    2008-01-01

    Mammalian fertilization is a complex multi-step process mediated by different molecules present on both gametes. CRISP1 (cysteine-rich secretory protein 1) is an epididymal protein thought to participate in gamete fusion through its binding to egg-complementary sites. Structure-function studies using recombinant fragments of CRISP1 as well as synthetic peptides reveal that its egg-binding ability resides in a 12 amino acid region corresponding to an evolutionary conserved motif of the CRISP family, named Signature 2 (S2). Further experiments analyzing both the ability of other CRISP proteins to bind to the rat egg and the amino acid sequence of their S2 regions show that the amino acid sequence of the S2 is needed for CRISP1 to interact with the egg. CRISP1 appears to be involved in the first step of sperm binding to the zona pellucida, identifying a novel role for this protein in fertilization. The observation that sperm testicular CRISP2 is also able to bind to the egg surface suggests a role for this protein in gamete fusion. Subsequent experiments confirmed the participation of CRISP2 in this step of fertilization and revealed that CRISP1 and CRISP2 interact with common egg surface binding sites. Together, these results suggest a functional cooperation between CRISP1 and CRISP2 to ensure the success of fertilization. These observations contribute to a better understanding of the molecular mechanisms underlying mammalian fertilization.

  8. XMAP310: A Xenopus Rescue-promoting Factor Localized to the Mitotic Spindle

    PubMed Central

    Andersen, Søren S.L.; Karsenti, Eric

    1997-01-01

    To understand the role of microtubule-associated proteins (MAPs) in the regulation of microtubule (MT) dynamics we have characterized MAPs prepared from Xenopus laevis eggs (Andersen, S.S.L., B. Buendia, J.E. Domínguez, A. Sawyer, and E. Karsenti. 1994. J. Cell Biol. 127:1289–1299). Here we report on the purification and characterization of a 310-kD MAP (XMAP310) that localizes to the nucleus in interphase and to mitotic spindle MTs in mitosis. XMAP310 is present in eggs, oocytes, a Xenopus tissue culture cell line, testis, and brain. We have purified XMAP310 to homogeneity from egg extracts. The purified protein cross-links pure MTs. Analysis of the effect of this protein on MT dynamics by time-lapse video microscopy has shown that it increases the rescue frequency 5–10-fold and decreases the shrinkage rate twofold. It has no effect on the growth rate or the catastrophe frequency. Microsequencing data suggest that XMAP230 and XMAP310 are novel MAPs. Although the three Xenopus MAPs characterized so far, XMAP215 (Vasquez, R.J., D.L. Gard, and L. Cassimeris. 1994. J. Cell Biol. 127:985–993), XMAP230, and XMAP310 are localized to the mitotic spindle, they have distinct effects on MT dynamics. While XMAP215 promotes rapid MT growth, XMAP230 decreases the catastrophe frequency and XMAP310 increases the rescue frequency. This may have important implications for the regulation of MT dynamics during spindle morphogenesis and chromosome segregation. PMID:9362515

  9. Glycosylated chicken ZP2 accumulates in the egg coat of immature oocytes and remains localized to the germinal disc region of mature eggs.

    PubMed

    Nishio, Shunsuke; Kohno, Yoshinori; Iwata, Yuki; Arai, Mayumi; Okumura, Hiroki; Oshima, Kenzi; Nadano, Daita; Matsuda, Tsukasa

    2014-11-01

    Vertebrate eggs are surrounded by an egg coat, which is a specific extracellular egg matrix consisting of several glycoproteins with a conserved zona pellucida (ZP) domain. Two mammalian egg coat subunits, ZP2 and ZP3, have been suggested to act as sperm receptors. In bird eggs, however, ZP2 has never been identified in the egg coat of mature oocytes and ovulated eggs. Here we report that chicken ZP2 is expressed in immature small follicles and remains as an egg-coat component locally in the germinal disc region of mature eggs. RT-PCR analysis indicated marked expression of the ZP2 and ZP4 genes in the granulosa cells of immature white follicles, whereas the ZP3 and ZPD genes showed marked expression in the cells of maturing yellow follicles. ZP2 was identified in the egg coat isolated from immature follicles as a heavily N-glycosylated glycoprotein of ∼200 kDa, which was enzymatically converted to a 70-kDa deglycosylated form. Immunoblotting and immunohistological analyses showed that ZP2 was localized around the germinal disc region of mature follicles. ZP2 was accumulated in the egg coat of immature white follicles at the earlier stages of oocyte development and became a minor component in the egg coat of maturing yellow follicles, except for the germinal disc region. Localization of ZP2 in the germinal disc region of mature eggs, where sperm bind to the egg coat at high density, suggests some role for ZP2 in the preferential binding and penetration of sperm in the germinal disc region of bird eggs. © 2014 by the Society for the Study of Reproduction, Inc.

  10. Early-life gut microbiome and egg allergy.

    PubMed

    Fazlollahi, M; Chun, Y; Grishin, A; Wood, R A; Burks, A W; Dawson, P; Jones, S M; Leung, D Y M; Sampson, H A; Sicherer, S H; Bunyavanich, S

    2018-07-01

    Gut microbiota may play a role in egg allergy. We sought to examine the association between early-life gut microbiota and egg allergy. We studied 141 children with egg allergy and controls from the multicenter Consortium of Food Allergy Research study. At enrollment (age 3 to 16 months), fecal samples were collected, and clinical evaluation, egg-specific IgE measurement, and egg skin prick test were performed. Gut microbiome was profiled by 16S rRNA sequencing. Analyses for the primary outcome of egg allergy at enrollment, and the secondary outcomes of egg sensitization at enrollment and resolution of egg allergy by age 8 years, were performed using Quantitative Insights into Microbial Ecology, Phylogenetic Investigation of Communities by Reconstruction of Unobserved States, and Statistical Analysis of Metagenomic Profiles. Compared to controls, increased alpha diversity and distinct taxa (PERMANOVA P = 5.0 × 10 -4 ) characterized the early-life gut microbiome of children with egg allergy. Genera from the Lachnospiraceae, Streptococcaceae, and Leuconostocaceae families were differentially abundant in children with egg allergy. Predicted metagenome functional analyses showed differential purine metabolism by the gut microbiota of egg-allergic subjects (Kruskal-Wallis P adj  = 0.021). Greater gut microbiome diversity and genera from Lachnospiraceae and Ruminococcaceae were associated with egg sensitization (PERMANOVA P = 5.0 × 10 -4 ). Among those with egg allergy, there was no association between early-life gut microbiota and egg allergy resolution by age 8 years. The distinct early-life gut microbiota in egg-allergic and egg-sensitized children identified by our study may point to targets for preventive or therapeutic intervention. © 2018 EAACI and John Wiley and Sons A/S. Published by John Wiley and Sons Ltd.

  11. Detection of Nosema bombycis by FTA cards and loop-mediated isothermal amplification (LAMP).

    PubMed

    Yan, Wei; Shen, Zhongyuan; Tang, Xudong; Xu, Li; Li, Qianlong; Yue, Yajie; Xiao, Shengyan; Fu, Xuliang

    2014-10-01

    We successfully established a detection method which exhibited a markedly higher sensitivity than previously developed detection methods for Nosema bombycis by combining glass beads, FTA card, and LAMP. Spores of N. bombycis were first broken by acid-washed glass beads; the DNA was subsequently extracted and purified with the FTA card, and LAMP was performed using primers (LSU296) designed based on the sequence of the LSU rRNA of N. bombycis. The minimum detection concentration was 10 spores/mL. When this method was used to detect pebrine disease in silkworm egg, the detection rate for 500 silkworm eggs, in which only one egg was infected with N. bombycis, was 100 % under our optimized conditions. If the number of eggs in the sample increased to 800 or 1,000, the sample was divided into two equal portions, and the eggs were smashed with glass beads after the addition of 1 mL of TE buffer. The liquid in two tubes was later mixed and applied to the FTA card, and the detection rates were 100 %. Furthermore, the LAMP method established in our study could detect N. bombycis infection in silkworm 24 h earlier than microscopy.

  12. Antiproliferative Activity of Egg Yolk Peptides in Human Colon Cancer Cells.

    PubMed

    Yousr, Marwa N; Aloqbi, Akram A; Omar, Ulfat M; Howell, Nazlin K

    2017-01-01

    Egg yolk peptides were successfully prepared from egg yolk protein by-products after lecithin extraction. Defatted egg yolk protein was hydrolyzed with pepsin and pancreatin and purified by gel filtration to produce egg yolk gel filtration fraction (EYGF-33) with antiproliferative activity. The highlight of this study was that the peptide EYGF-33 (1.0 mg/ml) significantly inhibits cell viability of colon cancer cells (Caco-2) with no inhibitory effects on the viability of human colon epithelial normal cells (HCEC) after 48 h. Reduced cell viability can be explained by cell cycle arrest in the S-phase in which DNA replication normally takes place. EYGF-33 significantly enhanced the production of superoxide anions in the mitochondria of Caco-2 cells; this could activate a mitochondrial apoptotic pathway leading to typical Poly Adenosine diphosphate-ribose polymerase (PARP) cleavage as observed in the Western blot result. The induction of apoptotic cell death by EYGF-33 was supported by the externalization of phosphatidylserine (PS). However, further elucidation of the mechanism of EYGF-33-mediated apoptosis would provide further support for its use as a potential therapeutic and chemopreventive agent.

  13. Vitamin D in the avian egg. Its molecular identity and mechanism of incorporation into yolk.

    PubMed Central

    Fraser, D R; Emtage, J S

    1976-01-01

    The chemical identity of vitamin D in the egg of the domestic fowl was studied by analysing radioactivity in eggs from hens injected with [3H]cholecalciferol. Labelled molecules were found throughout the egg, but the concentration of total radioactivity in albumin was only 5-7% of that in yolk. In lipid extracts of yolk, more than 90% of the radioactivity was as unchanged cholecalciferol and 5% as 25-hydroxycholecalciferol. Only about 3% of the radioactivity in albumin was chloroform-soluble, and of this 40% was 25-hydroxycholecalciferol and 15% was cholecalciferol. Evidence is presented to support the idea that the specific transport of cholecalciferol into yolk is mediated by a cholecalciferol-binding protein in blood. This protein forms a complex with yolk proteins in transit from liver to ovary via the blood. A cholecalciferol-binding protein, chromatographically similar to that from blood, was found in egg yolk. It is postulated that cholecalciferol forms part of a complex with its specific binding protein, Ca2+ and the yolk phosphoprotein, phosvitin. This complex is then incorporated into yolk by the thecal cells of the ovarian follicle. PMID:189757

  14. Food Modulation Controls Astaxanthin Accumulation in Eggs of the Sea Urchin Arbacia lixula.

    PubMed

    Galasso, Christian; Orefice, Ida; Toscano, Alfonso; Vega Fernández, Tomás; Musco, Luigi; Brunet, Christophe; Sansone, Clementina; Cirino, Paola

    2018-05-28

    The carotenoid astaxanthin has strong antioxidant properties with beneficial effects for various degenerative diseases. This carotenoid is produced by some microalgae species when cultivated in particular conditions, and, interestingly, it is a predominant carotenoid in aquatic animals throughout a broad range of taxa. Recently, astaxanthin was detected in the eggs of the sea urchin Arbacia lixula in relevant concentrations when this organism was maintained in culture. These results have paved the way for deeper research into astaxanthin production by this species, particularly in regards to how astaxanthin production can be modulated by diet. Results showed that the highest content of astaxanthin in eggs was observed in sea urchins fed on a diet enriched with Spirulina platensis . This result was confirmed by the high antioxidant activity recorded in the egg extracts of these animals. Our results suggest that (i) the sea urchin A. lixula is able to synthesize astaxanthin from precursors obtained from food, and (ii) it is possible to modulate the astaxanthin accumulation in sea urchin eggs by modifying the proportions of different food ingredients provided in their diet. This study demonstrates the large potential of sea urchin cultivation for the eco-sustainable production of healthy supplements for nutraceutical applications.

  15. Activation of amphibian oocytes by sperm extracts.

    PubMed

    Bonilla, F; Ajmat, M T; Sánchez Toranzo, G; Zelarayán, L; Oterino, J; Bühler, M I

    2008-11-01

    In the fertilization of most animals, egg activation is accompanied by an increase in cytoplasmatic Ca2+; however, the mechanism through which the fertilizing sperm induce this phenomenon is still controversial. An increase in intracellular free Ca2+ is required to trigger egg activation events, a process that includes cortical granule exocytosis, resumption and completion of meiosis and DNA replication, and culminates in the first mitotic cleavage. In this work, we investigated the effect of microinjection and incubation of different fractions of homologous sperm extract on the activation of Bufo arenarum oocytes matured in vitro. Two heat treatment-sensitive fractions obtained by chromatography were able to induce oocyte activation. The sperm fraction, which contained a 24 kDa protein, induced 90% activation when it was microinjected into the oocytes. Whilst the sperm fraction, which contained a 36 kDa protein, was able to induce about 70% activation only when it was applied on the oocyte surface.

  16. Initiation of DNA replication requires actin dynamics and formin activity.

    PubMed

    Parisis, Nikolaos; Krasinska, Liliana; Harker, Bethany; Urbach, Serge; Rossignol, Michel; Camasses, Alain; Dewar, James; Morin, Nathalie; Fisher, Daniel

    2017-11-02

    Nuclear actin regulates transcriptional programmes in a manner dependent on its levels and polymerisation state. This dynamics is determined by the balance of nucleocytoplasmic shuttling, formin- and redox-dependent filament polymerisation. Here, using Xenopus egg extracts and human somatic cells, we show that actin dynamics and formins are essential for DNA replication. In proliferating cells, formin inhibition abolishes nuclear transport and initiation of DNA replication, as well as general transcription. In replicating nuclei from transcriptionally silent Xenopus egg extracts, we identified numerous actin regulators, and disruption of actin dynamics abrogates nuclear transport, preventing NLS (nuclear localisation signal)-cargo release from RanGTP-importin complexes. Nuclear formin activity is further required to promote loading of cyclin-dependent kinase (CDK) and proliferating cell nuclear antigen (PCNA) onto chromatin, as well as initiation and elongation of DNA replication. Therefore, actin dynamics and formins control DNA replication by multiple direct and indirect mechanisms. © 2017 The Authors.

  17. Branching microtubule nucleation in Xenopus egg extracts mediated by augmin and TPX2

    PubMed Central

    Petry, Sabine; Groen, Aaron C.; Ishihara, Keisuke; Mitchison, Timothy J.; Vale, Ronald D.

    2013-01-01

    Summary The microtubules that comprise mitotic spindles in animal cells are nucleated at centrosomes and by spindle assembly factors that are activated in the vicinity of chromatin. Indirect evidence also has suggested that microtubules might be nucleated from pre-existing microtubules throughout the spindle, but this process has not been observed directly. Here, we demonstrate microtubule nucleation from the sides of existing microtubules in meiotic Xenopus egg extracts. Daughter microtubules grow at a low branch angle and with the same polarity as mother filaments. Branching microtubule nucleation requires gamma-tubulin and augmin and is stimulated by GTP-bound Ran and its effector TPX2, factors previously implicated in chromatin-stimulated nucleation. Because of the rapid amplification of microtubule numbers and the preservation of microtubule polarity, microtubule-dependent microtubule nucleation is well suited for spindle assembly and maintenance. PMID:23415226

  18. Autocatalytic microtubule nucleation determines the size and mass of Xenopus laevis egg extract spindles

    PubMed Central

    Decker, Franziska; Oriola, David; Dalton, Benjamin

    2018-01-01

    Regulation of size and growth is a fundamental problem in biology. A prominent example is the formation of the mitotic spindle, where protein concentration gradients around chromosomes are thought to regulate spindle growth by controlling microtubule nucleation. Previous evidence suggests that microtubules nucleate throughout the spindle structure. However, the mechanisms underlying microtubule nucleation and its spatial regulation are still unclear. Here, we developed an assay based on laser ablation to directly probe microtubule nucleation events in Xenopus laevis egg extracts. Combining this method with theory and quantitative microscopy, we show that the size of a spindle is controlled by autocatalytic growth of microtubules, driven by microtubule-stimulated microtubule nucleation. The autocatalytic activity of this nucleation system is spatially regulated by the limiting amounts of active microtubule nucleators, which decrease with distance from the chromosomes. This mechanism provides an upper limit to spindle size even when resources are not limiting. PMID:29323637

  19. Ovicidal and adulticidal potential of leaf and seed extract of Albizia lebbeck (L.) Benth. (Family: Fabaceae) against Culex quinquefasciatus, Aedes aegypti, and Anopheles stephensi (Diptera: Culicidae).

    PubMed

    Govindarajan, Marimuthu; Rajeswary, Mohan

    2015-05-01

    Several diseases are associated to the mosquito-human interaction. Mosquitoes are the carriers of severe and well-known illnesses such as malaria, arboviral encephalitis, dengue fever, chikungunya fever, West Nile virus, and yellow fever. These diseases produce significant morbidity and mortality in humans and livestock around the world. In the present study, hexane, benzene, chloroform, ethyl acetate, and methanol extracts of leaf and seed of Albizia lebbeck were assayed for their toxicity against three important vector mosquitoes, viz., Culex quinquefasciatus, Aedes aegypti, and Anopheles stephensi. The mean percent hatchability of the eggs was observed after 48 h post-treatment. The percent hatchability was inversely proportional to the concentration of extract and directly proportional to the eggs. All the five solvent extracts showed moderate ovicidal activity; however, the methanol extract showed the highest ovicidal activity. One hundred percent mortality was observed at 250, 200, and 150 ppm for leaf methanol extract and 375, 300, and 225 ppm for seed methanol extract of A. lebbeck against C. quinquefasciatus, Ae. aegypti, and An. stephensi, respectively. The adulticidal activity of plant leaf and seed extracts showed moderate toxic effect on the adult mosquitoes after 24 h of exposure period. However, the highest adulticidal activity was observed in the leaf methanol extract of A. lebbeck against An. stephensi where the LC₅₀ and LC₉₀ values were 65.12 and 117.70 ppm, respectively. Compared to leaf extracts, seeds have low potency against three mosquito species. No mortality was recorded in the control. Our data suggest that the crude hexane, benzene, chloroform, ethyl acetate, and methanol solvent extracts of A. lebbeck have the potential to be used as an eco-friendly approach for the control of the An. stephensi, Ae. aegypti, and C. quinquefasciatus. These results suggest that the leaf and seed extracts have the potential to be used as an ideal eco-friendly approach for the control of mosquitoes.

  20. Shifts in bacterial communities of eggshells and antimicrobial activities in eggs during incubation in a ground-nesting passerine.

    PubMed

    Grizard, Stéphanie; Versteegh, Maaike A; Ndithia, Henry K; Salles, Joana F; Tieleman, B Irene

    2015-01-01

    Microbial invasion of egg contents is a cause of embryonic death. To counter infection risks, the embryo is protected physically by the eggshell and chemically by antimicrobial proteins. If microbial pressure drives embryo mortality, then females may have evolved, through natural selection, to adapt their immune investment into eggs. Although frequently hypothesized, this match between immune allocation and microorganisms has not been explored yet. To examine if correlations between microbes on eggs and immunity in eggs exist, we collected eggs from red-capped larks (Calandrella cinerea) and simultaneously examined their bacterial communities and antimicrobial components--pH, lysozyme and ovotransferrin--during natural incubation. Using molecular techniques, we find that bacterial communities are highly dynamic: bacterial abundance increases from the onset to late incubation, Shannon's α-diversity index increases during early incubation stages, and β-diversity analysis shows that communities from 1 day-old clutches are phylogenetically more similar to each other than the older ones. Regarding the antimicrobials, we notice a decrease of pH and lysozyme concentration, while ovotransferrin concentration increases during incubation. Interestingly, we show that two eggs of the same clutch share equivalent immune protection, independent of clutch age. Lastly, our results provide limited evidence of significant correlation between antimicrobial compounds and bacterial communities. Our study examined simultaneously, for the first time in a wild bird, the dynamics of bacterial communities present on eggshells and of albumen-associated antimicrobial components during incubation and investigated their relationship. However, the link between microorganisms and immunity of eggs remains to be elucidated further. Identifying invading microbes and their roles in embryo mortality, as well as understanding the role of the eggshell microbiome, might be key to better understand avian strategies of immune maternal investment.

  1. Shifts in Bacterial Communities of Eggshells and Antimicrobial Activities in Eggs during Incubation in a Ground-Nesting Passerine

    PubMed Central

    Grizard, Stéphanie; Versteegh, Maaike A.; Ndithia, Henry K.; Salles, Joana F.; Tieleman, B. Irene

    2015-01-01

    Microbial invasion of egg contents is a cause of embryonic death. To counter infection risks, the embryo is protected physically by the eggshell and chemically by antimicrobial proteins. If microbial pressure drives embryo mortality, then females may have evolved, through natural selection, to adapt their immune investment into eggs. Although frequently hypothesized, this match between immune allocation and microorganisms has not been explored yet. To examine if correlations between microbes on eggs and immunity in eggs exist, we collected eggs from red-capped larks (Calandrella cinerea) and simultaneously examined their bacterial communities and antimicrobial components—pH, lysozyme and ovotransferrin—during natural incubation. Using molecular techniques, we find that bacterial communities are highly dynamic: bacterial abundance increases from the onset to late incubation, Shannon’s α-diversity index increases during early incubation stages, and β-diversity analysis shows that communities from 1 day-old clutches are phylogenetically more similar to each other than the older ones. Regarding the antimicrobials, we notice a decrease of pH and lysozyme concentration, while ovotransferrin concentration increases during incubation. Interestingly, we show that two eggs of the same clutch share equivalent immune protection, independent of clutch age. Lastly, our results provide limited evidence of significant correlation between antimicrobial compounds and bacterial communities. Our study examined simultaneously, for the first time in a wild bird, the dynamics of bacterial communities present on eggshells and of albumen-associated antimicrobial components during incubation and investigated their relationship. However, the link between microorganisms and immunity of eggs remains to be elucidated further. Identifying invading microbes and their roles in embryo mortality, as well as understanding the role of the eggshell microbiome, might be key to better understand avian strategies of immune maternal investment. PMID:25880684

  2. Positional cloning of a Bombyx pink-eyed white egg locus reveals the major role of cardinal in ommochrome synthesis

    PubMed Central

    Osanai-Futahashi, M; Tatematsu, K-i; Futahashi, R; Narukawa, J; Takasu, Y; Kayukawa, T; Shinoda, T; Ishige, T; Yajima, S; Tamura, T; Yamamoto, K; Sezutsu, H

    2016-01-01

    Ommochromes are major insect pigments involved in coloration of compound eyes, eggs, epidermis and wings. In the silkworm Bombyx mori, adult compound eyes and eggs contain a mixture of the ommochrome pigments such as ommin and xanthommatin. Here, we identified the gene involved in ommochrome biosynthesis by positional cloning of B. mori egg and eye color mutant pink-eyed white egg (pe). The recessive homozygote of pe has bright red eyes and white or pale pink eggs instead of a normal dark coloration due to the decrease of dark ommochrome pigments. By genetic linkage analysis, we narrowed down the pe-linked region to ~258 kb, containing 17 predicted genes. RNA sequencing analyses showed that the expression of one candidate gene, the ortholog of Drosophila haem peroxidase cardinal, coincided with egg pigmentation timing, similar to other ommochrome-related genes such as Bm-scarlet and Bm-re. In two pe strains, a common missense mutation was found within a conserved motif of B. mori cardinal homolog (Bm-cardinal). RNA interference-mediated knockdown and transcription activator-like effector nuclease (TALEN)-mediated knockout of the Bm-cardinal gene produced the same phenotype as pe in terms of egg, adult eye and larval epidermis coloration. A complementation test of the pe mutant with the TALEN-mediated Bm-cardinal-deficient strain showed that the mutant phenotype could not be rescued, indicating that Bm-cardinal is responsible for pe. Moreover, knockdown of the cardinal homolog in Tribolium castaneum also induced red compound eyes. Our results indicate that cardinal plays a major role in ommochrome synthesis of holometabolous insects. PMID:26328757

  3. Positional cloning of a Bombyx pink-eyed white egg locus reveals the major role of cardinal in ommochrome synthesis.

    PubMed

    Osanai-Futahashi, M; Tatematsu, K-I; Futahashi, R; Narukawa, J; Takasu, Y; Kayukawa, T; Shinoda, T; Ishige, T; Yajima, S; Tamura, T; Yamamoto, K; Sezutsu, H

    2016-02-01

    Ommochromes are major insect pigments involved in coloration of compound eyes, eggs, epidermis and wings. In the silkworm Bombyx mori, adult compound eyes and eggs contain a mixture of the ommochrome pigments such as ommin and xanthommatin. Here, we identified the gene involved in ommochrome biosynthesis by positional cloning of B. mori egg and eye color mutant pink-eyed white egg (pe). The recessive homozygote of pe has bright red eyes and white or pale pink eggs instead of a normal dark coloration due to the decrease of dark ommochrome pigments. By genetic linkage analysis, we narrowed down the pe-linked region to ~258 kb, containing 17 predicted genes. RNA sequencing analyses showed that the expression of one candidate gene, the ortholog of Drosophila haem peroxidase cardinal, coincided with egg pigmentation timing, similar to other ommochrome-related genes such as Bm-scarlet and Bm-re. In two pe strains, a common missense mutation was found within a conserved motif of B. mori cardinal homolog (Bm-cardinal). RNA interference-mediated knockdown and transcription activator-like effector nuclease (TALEN)-mediated knockout of the Bm-cardinal gene produced the same phenotype as pe in terms of egg, adult eye and larval epidermis coloration. A complementation test of the pe mutant with the TALEN-mediated Bm-cardinal-deficient strain showed that the mutant phenotype could not be rescued, indicating that Bm-cardinal is responsible for pe. Moreover, knockdown of the cardinal homolog in Tribolium castaneum also induced red compound eyes. Our results indicate that cardinal plays a major role in ommochrome synthesis of holometabolous insects.

  4. CD9 tetraspanin generates fusion competent sites on the egg membrane for mammalian fertilization

    PubMed Central

    Jégou, Antoine; Ziyyat, Ahmed; Barraud-Lange, Virginie; Perez, Eric; Wolf, Jean Philippe; Pincet, Frédéric; Gourier, Christine

    2011-01-01

    CD9 tetraspanin is the only egg membrane protein known to be essential for fertilization. To investigate its role, we have measured, on a unique acrosome reacted sperm brought in contact with an egg, the adhesion probability and strength with a sensitivity of a single molecule attachment. Probing the binding events at different locations of wild-type egg we described different modes of interaction. Here, we show that more gamete adhesion events occur on Cd9 null eggs but that the strongest interaction mode disappears. We propose that sperm–egg fusion is a direct consequence of CD9 controlled sperm–egg adhesion properties. CD9 generates adhesion sites responsible for the strongest of the observed gamete interaction. These strong adhesion sites impose, during the whole interaction lifetime, a tight proximity of the gamete membranes, which is a requirement for fusion to take place. The CD9-induced adhesion sites would be the actual location where fusion occurs. PMID:21690351

  5. Early plant defence against insect attack: involvement of reactive oxygen species in plant responses to insect egg deposition.

    PubMed

    Bittner, Norbert; Trauer-Kizilelma, Ute; Hilker, Monika

    2017-05-01

    Pinus sylvestris responds to insect egg deposition by ROS accumulation linked with reduced activity of the ROS scavenger catalase. Egg mortality in needles with hypersensitive response (HR)-like symptoms is enhanced. Aggressive reactive oxygen species (ROS) play an important role in plant defence against biotic stressors, including herbivorous insects. Plants may even generate ROS in response to insect eggs, thus effectively fighting against future larval herbivory. However, so far nothing is known on how ROS-mediated plant defence against insect eggs is enzymatically regulated. Neither do we know how insects cope with egg-induced plant ROS. We addressed these gaps of knowledge by studying the activities of ROS-related enzymes in Pinus sylvestris deposited with eggs of the herbivorous sawfly Diprion pini. This species cuts a slit into pine needles and inserts its eggs into the needle tissue. About a quarter of egg-deposited needles show chlorotic tissue at the oviposition sites, indicating hypersensitive response-like direct defence responses resulting in reduced larval hatching from eggs. Hydrogen peroxide and peroxidase sensitive staining of sections of egg-deposited pine needles revealed the presence of hydrogen peroxide and peroxidase activity in needle tissue close to the eggs. Activity of ROS-producing NADPH-oxidase did not increase after egg deposition. However, the activity of the ROS-detoxifying enzyme catalase decreased after egg deposition and ovipositional wounding of needles. These results show that local ROS accumulation at the oviposition site is not caused by increased NADPH-oxidase activity, but reduced activity of pine needle catalase may contribute to it. However, our data suggest that pine sawflies can counteract the egg deposition-induced hydrogen peroxide accumulation in pine needles by high catalase activity in their oviduct secretion which is released with the eggs into pine tissue.

  6. Geographical Variation in Egg Mass and Egg Content in a Passerine Bird

    PubMed Central

    Ruuskanen, Suvi; Siitari, Heli; Eeva, Tapio; Belskii, Eugen; Järvinen, Antero; Kerimov, Anvar; Krams, Indrikis; Moreno, Juan; Morosinotto, Chiara; Mänd, Raivo; Möstl, Erich; Orell, Markku; Qvarnström, Anna; Salminen, Juha-Pekka; Slater, Fred; Tilgar, Vallo; Visser, Marcel E.; Winkel, Wolfgang; Zang, Herwig; Laaksonen, Toni

    2011-01-01

    Reproductive, phenotypic and life-history traits in many animal and plant taxa show geographic variation, indicating spatial variation in selection regimes. Maternal deposition to avian eggs, such as hormones, antibodies and antioxidants, critically affect development of the offspring, with long-lasting effects on the phenotype and fitness. Little is however known about large-scale geographical patterns of variation in maternal deposition to eggs. We studied geographical variation in egg components of a passerine bird, the pied flycatcher (Ficedula hypoleuca), by collecting samples from 16 populations and measuring egg and yolk mass, albumen lysozyme activity, yolk immunoglobulins, yolk androgens and yolk total carotenoids. We found significant variation among populations in most egg components, but ca. 90% of the variation was among individuals within populations. Population however explained 40% of the variation in carotenoid levels. In contrast to our hypothesis, we found geographical trends only in carotenoids, but not in any of the other egg components. Our results thus suggest high within-population variation and leave little scope for local adaptation and genetic differentiation in deposition of different egg components. The role of these maternally-derived resources in evolutionary change should be further investigated. PMID:22110579

  7. Risk factors for the occurrence of sporadic Salmonella enterica serotype enteritidis infections in children in France: a national case-control study.

    PubMed Central

    Delarocque-Astagneau, E.; Desenclos, J. C.; Bouvet, P.; Grimont, P. A.

    1998-01-01

    To determine risk factors associated with the occurrence of sporadic cases of Salmonella enteritidis infections among children in France, we conducted a matched case-control study. Cases were identified between 1 March and 30 September 1995. One hundred and five pairs of cases and controls matched for age and place of residence were interviewed. In the 1-5 years age group, illness was associated with the consumption of raw eggs or undercooked egg-containing foods (OR 2.4, 95% CI 1.2-4.8). Storing eggs more than 2 weeks after purchase was associated with Salmonella enteritidis infection (OR 3.8, 95% CI 1.4-10.2), particularly during the summer period (OR 6.0, 95% CI 1.3-26.8). Cases were more likely to report a case of diarrhoea in the household 10-3 days before the onset of symptoms, particularly in the age group < or = 1 year (P = 0.01). This study confirms the link between eggs and the occurrence of sporadic cases of Salmonella enteritidis among children, highlights the potential role of prolonged egg storage and underlines the role of person-to-person transmission in infants. PMID:10030705

  8. Calcium-responsive contractility during fertilization in sea urchin eggs.

    PubMed

    Stack, Christianna; Lucero, Amy J; Shuster, Charles B

    2006-04-01

    Fertilization triggers a reorganization of oocyte cytoskeleton, and in sea urchins, there is a dramatic increase in cortical F-actin. However, the role that myosin II plays during fertilization remains largely unexplored. Myosin II is localized to the cortical cytoskeleton both before and after fertilization and to examine myosin II contractility in living cells, Lytechinus pictus eggs were observed by time-lapse microscopy. Upon sperm binding, a cell surface deflection traversed the egg that was followed by and dependent on the calcium wave. The calcium-dependence of surface contractility could be reproduced in unfertilized eggs, where mobilization of intracellular calcium in unfertilized eggs under compression resulted in a marked contractile response. Lastly, inhibition of myosin II delayed absorption of the fertilization cone, suggesting that myosin II not only responds to the same signals that activate eggs but also participates in the remodeling of the cortical actomyosin cytoskeleton during the first zygotic cell cycle. (c) 2006 Wiley-Liss, Inc.

  9. Calcium-Responsive Contractility During Fertilization in Sea Urchin Eggs

    PubMed Central

    Stack, Christianna; Lucero, Amy J.; Shuster, Charles B.

    2008-01-01

    Fertilization triggers a reorganization of oocyte cytoskeleton, and in sea urchins there is a dramatic increase in cortical F-actin. However, the role that myosin II plays during fertilization remains largely unexplored. Myosin II is localized to the cortical cytoskeleton both prior to- and following fertilization, and to examine myosin II contractility in living cells, Lytechinus pictus eggs were observed by time-lapse microscopy. Upon sperm binding, a cell surface deflection traversed the egg that was followed- and dependent on the calcium wave. The calcium-dependence of surface contractility could be reproduced in unfertilized eggs, where mobilization of intracellular calcium in unfertilized eggs under compression resulted in a marked contractile response. Lastly, inhibition of myosin II delayed absorption of the fertilization cone, suggesting that myosin II not only responds to the same signals that activate eggs, but also participates in the remodeling of the cortical actomyosin cytoskeleton during the first zygotic cell cycle. PMID:16470603

  10. Comparison of the in vitro anthelmintic effects of Acacia nilotica and Acacia raddiana

    PubMed Central

    Zabré, Geneviève; Kaboré, Adama; Bayala, Balé; Katiki, Luciana M.; Costa-Júnior, Lívio Martins; Tamboura, Hamidou H.; Belem, Adrien M.G.; Abdalla, Adibe L.; Niderkorn, Vincent; Hoste, Hervé; Louvandini, Helder

    2017-01-01

    Gastrointestinal nematodes are a major threat to small ruminant rearing in the Sahel area, where farmers traditionally use bioactive plants to control these worms, including Acacia nilotica and Acacia raddiana. The main aim of this study was to screen the potential anthelmintic properties of aqueous and acetone extracts of leaves of these two plants based on three in vitro assays: (1) the egg hatch inhibition assay (EHA); (2) the larvae exsheathment inhibition assay (LEIA) using Haemonchus contortus as a model; and (3) an adult mortality test (AMT) applied on Caenorhabditis elegans. For the EHA, only A. raddiana was effective with IC50 = 1.58 mg/mL for aqueous extract, and IC50 = 0.58 mg/mL for acetonic extract. For the LEIA, all extracts inhibited the exsheathment of larvae compared to the controls, and the aqueous extract of A. nilotica was more larvicidal with IC50 = 0.195 mg/mL. In general, all responses to the substances were dose-dependent and were significantly different from the control group (p < 0.05). For the AMT, the extracts of the two Acacia species were effective but A. raddiana showed greater efficacy with 100% mortality at 2.5 mg/mL and LC50 = 0.84 mg/mL (acetonic extract). The addition of polyvinyl polypyrrolidone (PVPP) to the extracts suggested that tannins were responsible for blocking egg eclosion and inducing adult mortality but were not responsible for exsheathment inhibition. These results suggest that the leaves of these Acacia species possess ovicidal and larvicidal activities in vitro against H. contortus, and adulticidal effects against C. elegans. PMID:29173278

  11. Vitellogenin knockdown strongly affects cotton boll weevil egg viability but not the number of eggs laid by females.

    PubMed

    Coelho, Roberta R; de Souza Júnior, José Dijair Antonino; Firmino, Alexandre A P; de Macedo, Leonardo L P; Fonseca, Fernando C A; Terra, Walter R; Engler, Gilbert; de Almeida Engler, Janice; da Silva, Maria Cristina M; Grossi-de-Sa, Maria Fatima

    2016-09-01

    Vitellogenin (Vg), a yolk protein precursor, is the primary egg nutrient source involved in insect reproduction and embryo development. The Cotton Boll weevil (CBW) Anthonomus grandis Boheman, the most important cotton pest in Americas, accumulates large amounts of Vg during reproduction. However, the precise role of this protein during embryo development in this insect remains unknown. Herein, we investigated the effects of vitellogenin (AgraVg) knockdown on the egg-laying and egg viability in A. grandis females, and also characterized morphologically the unviable eggs. AgraVg transcripts were found during all developmental stages of A. grandis, with highest abundance in females. Silencing of AgraVg culminated in a significant reduction in transcript amount, around 90%. Despite this transcriptional reduction, egg-laying was not affected in dsRNA-treated females but almost 100% of the eggs lost their viability. Eggs from dsRNA-treated females showed aberrant embryos phenotype suggesting interference at different stages of embryonic development. Unlike for other insects, the AgraVg knockdown did not affect the egg-laying ability of A. grandis, but hampered A. grandis reproduction by perturbing embryo development. We concluded that the Vg protein is essential for A. grandis reproduction and a good candidate to bio-engineer the resistance against this devastating cotton pest.

  12. Organochlorine pesticides, PCBs, trace elements and metals in western pond turtle eggs from Oregon

    USGS Publications Warehouse

    Henny, Charles J.; Beal, K.F.; Bury, R. Bruce; Goggans, R.

    2003-01-01

    With increased concern over the status of reptile populations globally, contaminant studies should be part of species evaluations. We analyzed eggs of western pond turtles from Fern Ridge Reservoir in western Oregon for 20 organochlorine (OC) pesticides or metabolites, 42 congener-specific polychlorinated biphenyls (PCBs), and 16 trace elements or metals. These eggs represent the first of this species analyzed. The OC pesticides and PCB residue concentrations in the western pond turtle eggs were generally low and similar to those found in eggs of snapping turtles from a remote site in Ontario, Canada. Western pond turtle eggs also contained mercury and chromium, which are metals of special concern. Although few reptilian eggs have been analyzed for metals, the 44.9 mug/g dry weight chromium in a western pond turtle egg in this study may be the highest reported in a reptilian egg. We found no significant difference in contaminant concentrations in eggs from nests in Oregon, where all turtle eggs failed to hatch compared to those where some eggs hatched. During this initial project, however, we were unable to assess fully the role of OCs, PCBs and other contaminants in the western pond turtle decline. Factors other than contaminants may be involved. In another study, snapping turtle eggs near the Great Lakes-St. Lawrence River basin were much more contaminated with evidence reported of effects on sex differentiation and reproductive endocrine function. Egg hatchability, the only reproductive parameter monitored, may not be the most sensitive endpoint. Other endpoints, including endocrine function, deformity rates, growth rates, and sex determination need study.

  13. Impact of potassium bromate and potassium iodate in a pound cake system.

    PubMed

    Wilderjans, Edith; Lagrain, Bert; Brijs, Kristof; Delcour, Jan A

    2010-05-26

    This study investigates the impact of the oxidants potassium bromate and potassium iodate (8, 16, 32, 64, and 128 micromol/g dry matter of egg white protein) on pound cake making. The impact of the oxidants on egg white characteristics was studied in a model system. Differential scanning calorimetry showed that the oxidants caused egg white to denature later. During heating in a rapid visco analyzer, the oxidants caused the free sulfhydryl (SH) group levels to decrease more intensively and over a smaller temperature range. The oxidants made the proteins more resistant to decreases in protein extractability in sodium dodecyl sulfate containing buffer during cake recipe mixing and less resistant to such decreases during cake baking. We assume that, during baking, the degree to which SH/disulfide exchange and SH oxidation can occur depends on the properties of the protein at the onset of the process. In our view, the prevention of extractability loss during mixing increased the availability of SH groups and caused more such loss during baking. During cooling, all cakes baked with added oxidants showed less collapse. On the basis of the presented data, we put forward that only those protein reactions that occur during baking contribute to the formation of a network that supports final cake structure and prevents collapse.

  14. Mlh1 is required for female fertility in Drosophila melanogaster: An outcome of effects on meiotic crossing over, ovarian follicles and egg activation.

    PubMed

    Vimal, Divya; Kumar, Saurabh; Pandey, Ashutosh; Sharma, Divya; Saini, Sanjay; Gupta, Snigdha; Ravi Ram, Kristipati; Chowdhuri, Debapratim Kar

    2018-03-01

    Mismatch repair (MMR) system, a conserved DNA repair pathway, plays crucial role in DNA recombination and is involved in gametogenesis. The impact of alterations in MMR family of proteins (bacterial MutS and MutL homologues) on mammalian fertility is well documented. However, an insight to the role of MMR in reproduction of non-mammalian organisms is limited. Hence, in the present study, we analysed the impact of mlh1 (a MutL homologue) on meiotic crossing over/recombination and fertility in a genetically tractable model, Drosophila melanogaster. Using mlh1 e00130 hypomorphic allele, we report female specific adverse reproductive outcome for reduced mlh1 in Drosophila: mlh1 e00130 homozygous females had severely reduced fertility while males were fertile. Further, mlh1 e00130 females contained small ovaries with large number of early stages as well as significantly reduced mature oocytes, and laid fewer eggs, indicating discrepancies in egg production and ovulation. These observations contrast the sex independent and/or male specific sterility and normal follicular development as well as ovulation reported so far for MMR family proteins in mammals. However, analogous to the role(s) of mlh1 in meiotic crossing over and DNA repair processes underlying mammalian fertility, ovarian follicles from mlh1 e00130 females contained significantly increased DNA double strand breaks (DSBs) and reduced synaptonemal complex foci. In addition, large proportion of fertilized eggs display discrepancies in egg activation and fail to proceed beyond stage 5 of embryogenesis. Hence, reduction of the Mlh1 protein level leads to defective oocytes that fail to complete embryogenesis after fertilization thereby reducing female fertility. Copyright © 2017 Elsevier GmbH. All rights reserved.

  15. The influence of ph and waterborne metals on egg fertilization of the blue mussel (Mytilus edulis), the oyster (Crassostrea gigas) and the sea urchin (Paracentrotus lividus).

    PubMed

    Riba, Inmaculada; Gabrielyan, Bardukh; Khosrovyan, Alla; Luque, Angel; Del Valls, T Angel

    2016-07-01

    This study evaluated the combined effect of pH and metals on the egg fertilization process of two estuarine species, the blue mussel (Mytilus edulis), the oyster (Crassostrea gigas) and a marine species, the sea urchin (Paracentrotus lividus). The success of egg fertilization was examined after exposure of gametes to sediment extracts of various degrees of contamination at pH 6.0, 6.5, 7.0, 7.5 and 8.0. At the pH levels from 6.5 to 8.0, the egg fertilization of the different species demonstrated different sensitivity to metal and/or acidic exposure. In all species, the results revealed that egg fertilization was almost completely inhibited at pH 6.0. The egg fertilization of the blue mussel M. edulis was the least sensitive to the exposure while that of the sea urchin P. lividus demonstrated a concentration-dependent response to the pH levels from 6.5 to 8.0. The results of this study revealed that acidity increased the concentration of several metal ions (Cr, Ni, Cu, Zn, Cd, and Pb) but reduced its availability to the organisms, probably related to the reactivity of the ions with most non-metals or to the competition among metals and other waterborne constituents.

  16. Quantification of oxysterols in Dutch foods: egg products and mixed diets.

    PubMed

    van de Bovenkamp, P; Kosmeijer-Schuil, T G; Katan, M B

    1988-11-01

    A sensitive and specific method is described for quantifying various cholesterol oxidation products in foodstuffs, including 7 beta-hydroxycholesterol, cholesterol-alpha-epoxide, cholestane-triol, 7-ketocholesterol and 25-hydroxycholesterol. A chloroform-methanol extract of the food was fractionated over two successive silica columns. Two fractions containing different classes of oxysterols were then analyzed as trimethylsilyl derivatives by capillary gas liquid chromatography, using on-column injection and a temperature gradient from 70 to 200 degrees C. The detection limit was about 0.5 microgram/g dry weight for egg yolk powder. Fresh egg yolk contained only 1.2 micrograms/g of total oxides per g dry weight, showing that artifactual oxidation during the procedure was minimal. Recovery of 5 pure oxysterols added to egg yolk at levels of 6.5 and 10 micrograms/g was between 93 and 102%. In commercial egg yolk and whole egg powder stored for one year, total amounts of oxysterols ranging from 21 to 137 micrograms/g dry weight were found. In duplicates of mixed Dutch diets, total amounts ranged from 3.6 to 6.2 micrograms/g dry weight. Duplicates containing mostly fried and baked foods did not have higher levels than duplicates in which foods had been prepared by boiling or left raw. We conclude that a normal mixed diet provides only minor amounts of cholesterol oxidation products.

  17. Calibrating compound-specific stable nitrogen isotope analysis in avian eggs as bioarchives for paleoreconstruction

    NASA Astrophysics Data System (ADS)

    Preciado, C., Jr.

    2016-12-01

    Compound-specific stable isotope analysis (CSIA) has become a powerful tool for reconstructing consumer-resource relationships in modern and ancient systems. Stable nitrogen isotope analysis (δ15N) of "trophic" and "source" amino acids provides independent proxies of trophic position and the δ15N value at the base of the food web, respectively. When applied to avian egg tissues (e.g., shell protein, membrane), which can be preserved in the archaeological record, this approach can be used to address complex questions in food web architecture and biogeochemical cycling. In this study, we examined how sample-processing protocols affected the chromatography and reliability of δ15N values of individual amino acids in avian (chicken and penguin) egg components (shell protein, membrane, yolk, and albumen) via gas chromatography-combustion-isotope ratio mass spectrometry. "Unprocessed" egg tissues underwent standard acid hydrolysis protocols prior to derivatization, and resulted in poor chromatography with highly variable δ15N values across replicates. "Processed" samples were eluted through cation exchange columns (Dowex 50WX*400) prior to derivatization, followed by a P-buffer (KH2PO4 + Na2HPO4)-chloroform centrifugation extraction to remove confounding peaks and matrix impurities. These additional procedures greatly improved the chromatography of the "processed" samples, revealing better peak separation and baseline integration as well as lower δ15N variability. Additionally, a standard lipid extraction was necessary for yolk but not membrane, albumen, and shell. While the additional procedures applied to the "columned" samples did result in a significant reduction in sample yield ( 20%), it was non-fractionating and thus only affected the total sample sizes necessary for δ15N CSIA (shell protein 50mg and membrane, yolk, and albumen 0.5mg). The protocols developed here will streamline CSIA of egg tissues for future work in avian ecology.

  18. A Targeted LC-MS/MS Method for the Simultaneous Detection and Quantitation of Egg, Milk, and Peanut Allergens in Sugar Cookies.

    PubMed

    Boo, Chelsea C; Parker, Christine H; Jackson, Lauren S

    2018-01-01

    Food allergy is a growing public health concern, with many individuals reporting allergies to multiple food sources. Compliance with food labeling regulations and prevention of inadvertent cross-contact in manufacturing requires the use of reliable methods for the detection and quantitation of allergens in processed foods. In this work, a novel liquid chromatography-tandem mass spectrometry multiple-reaction monitoring method for multiallergen detection and quantitation of egg, milk, and peanut was developed and evaluated in an allergen-incurred baked sugar cookie matrix. A systematic evaluation of method parameters, including sample extraction, concentration, and digestion, were optimized for candidate allergen peptide markers. The optimized method enabled the reliable detection and quantitation of egg, milk, and peanut allergens in sugar cookies, with allergen concentrations as low as 5 ppm allergen-incurred ingredient.

  19. Fertilization and development of eggs of the South African clawed toad, Xenopus laevis, on sounding rockets in space

    NASA Astrophysics Data System (ADS)

    Ubbels, Geertje A.; Berendsen, Willem; Kerkvliet, Sonja; Narraway, Jenny

    Egg rotation and centrifugation experiments strongly suggest a role for gravity in the determination of the spatial structure of amphibian embryos. Decisive experiments can only be made in Space. Eggs of Xenopus laevis, the South African clawed toad, were the first vertebrate eggs which were successfully fertilized on Sounding Rockets in Space. Unfixed, newly fertilized eggs survived reentry, and a reasonable number showed a seemingly normal gastrulation but died between gastrulation and neurulation. Only a few reached the larval stage, but these developed abnormally. In the future, we inted to test whether this abnormal morphogenesis is due to reentry perturbations, or due to a real microgravity effect, through perturbation of the reinitiation of meiosis and other processes, or started by later sperm penetration.

  20. Fertilization and development of eggs of the South African clawed toad, Xenopus laevis, on sounding rockets in space.

    PubMed

    Ubbels, G A; Berendsen, W; Kerkvliet, S; Narraway, J

    1992-01-01

    Egg rotation and centrifugation experiments strongly suggest a role for gravity in the determination of the spatial structure of amphibian embryos. Decisive experiments can only be made in Space. Eggs of Xenopus laevis, the South African clawed toad, were the first vertebrate eggs which were successfully fertilized on Sounding Rockets in Space. Unfixed, newly fertilized eggs survived reentry, and a reasonable number showed a seemingly normal gastrulation but died between gastrulation and neurulation. Only a few reached the larval stage, but these developed abnormally. In the future, we intend to test whether this abnormal morphogenesis is due to reentry perturbations, or due to a real microgravity effect, through perturbation of the reinitiation of meiosis and other processes, or started by later sperm penetration.

  1. Administration of yellow fever vaccine in patients with egg allergy.

    PubMed

    Rutkowski, K; Ewan, P W; Nasser, S M

    2013-01-01

    The population of large parts of Africa, South America and travellers to these areas are at risk of yellow fever (YF) with a 50% mortality risk. Yellow fever vaccine (YFV) propagated in hens' eggs confers protection in 95% of the vaccinated. The rate of anaphylaxis for YFV ranges from 0.42 to 1.8/100,000 doses with most cases considered to be due to egg allergy. Egg allergy is a contraindication for the YFV. Nevertheless, the potential fatal sequelae from YF give the incentive to protect everyone at risk irrespective of their allergic status. Six subjects who had had a recent reaction to egg and who were travelling to endemic areas (3 adults and 3 children) underwent skin prick tests (SPT) with undiluted YFV and egg extract. Intradermal tests for YFV were undertaken at a 1:10 dilution. In 4 egg-allergic patients with a positive SPT to YFV, a 7-step desensitization protocol was used. A 2-step (10 + 90%) protocol was used in the 2 subjects with a negative YFV SPT. Premedication was not administered. All 6 patients were successfully vaccinated. Four patients completed desensitization: 1 developed mild local erythema at the injection site, 1 had fleeting generalized urticaria with local erythema/angioedema and 2 did not experience any adverse reactions. Patients who received YFV in 2 steps developed no adverse reactions. We describe the successful administration of YFV in 6 egg-allergic patients. The Cambridge Allergy 7-step protocol allows for its safe administration in patients with positive SPT to YFV. A 2-step protocol can be used in patients with negative YFV SPT. Copyright © 2013 S. Karger AG, Basel.

  2. Replication, checkpoint suppression and structure of centromeric DNA

    PubMed Central

    Romeo, Francesco; Costanzo, Vincenzo

    2016-01-01

    ABSTRACT Human centromeres contain large amounts of repetitive DNA sequences known as α satellite DNA, which can be difficult to replicate and whose functional role is unclear. Recently, we have characterized protein composition, structural organization and checkpoint response to stalled replication forks of centromeric chromatin reconstituted in Xenopus laevis egg extract. We showed that centromeric DNA has high affinity for SMC2-4 subunits of condensins and for CENP-A, it is enriched for DNA repair factors and suppresses the ATR checkpoint to ensure its efficient replication. We also showed that centromeric chromatin forms condensins enriched and topologically constrained DNA loops, which likely contribute to the overall structure of the centromere. These findings have important implications on how chromosomes are organized and genome stability is maintained in mammalian cells. PMID:27893298

  3. Eggs: The role of gravity in the establishment of the dorso-ventral axis in the amphibian embryo (7-IML-1)

    NASA Technical Reports Server (NTRS)

    Ubbels, Geertje A.

    1992-01-01

    The purpose of this experiment is to fertilize frog (Xenopus laevis) eggs under microgravity, to perform histological fixations at two different programmed times, and after return to Earth, to determine whether timing and pattern of egg cleavages and axis formation are normal. Because of the limited viability of the gametes, this experiment will be the very first to be activated in the Biorack. Various aspects of this investigation are discussed.

  4. Ancient Mitochondrial DNA Analyses of Ascaris Eggs Discovered in Coprolites from Joseon Tomb

    PubMed Central

    Oh, Chang Seok; Seo, Min; Hong, Jong Ha; Chai, Jong-Yil; Oh, Seung Whan; Park, Jun Bum; Shin, Dong Hoon

    2015-01-01

    Analysis of ancient DNA (aDNA) extracted from Ascaris is very important for understanding the phylogenetic lineage of the parasite species. When aDNAs obtained from a Joseon tomb (SN2-19-1) coprolite in which Ascaris eggs were identified were amplified with primers for cytochrome b (cyt b) and 18S small subunit ribosomal RNA (18S rRNA) gene, the outcome exhibited Ascaris specific amplicon bands. By cloning, sequencing, and analysis of the amplified DNA, we obtained information valuable for comprehending genetic lineage of Ascaris prevalent among pre-modern Joseon peoples. PMID:25925186

  5. 7 CFR 51.1179 - Method of juice extraction.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... Agriculture Regulations of the Department of Agriculture AGRICULTURAL MARKETING SERVICE (Standards, Inspections, Marketing Practices), DEPARTMENT OF AGRICULTURE REGULATIONS AND STANDARDS UNDER THE AGRICULTURAL MARKETING ACT OF 1946 AND THE EGG PRODUCTS INSPECTION ACT FRESH FRUITS, VEGETABLES AND OTHER PRODUCTS 1 2...

  6. 7 CFR 51.1179 - Method of juice extraction.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... Agriculture Regulations of the Department of Agriculture AGRICULTURAL MARKETING SERVICE (Standards, Inspections, Marketing Practices), DEPARTMENT OF AGRICULTURE REGULATIONS AND STANDARDS UNDER THE AGRICULTURAL MARKETING ACT OF 1946 AND THE EGG PRODUCTS INSPECTION ACT FRESH FRUITS, VEGETABLES AND OTHER PRODUCTS 1,2...

  7. Role of low density lipoprotein in the activation of plasma lysolecithin acyltransferase activity. Effect of chemical and enzymatic modifications of the lipoprotein on enzyme activity.

    PubMed

    Subbaiah, P V; Chen, C H; Bagdade, J D; Albers, J J

    1985-01-01

    The effect of various chemical and enzymatic modifications of low density lipoprotein (LDL) on its ability to activate the isolated human plasma lysolecithin acyltransferase (LAT) was studied. Removal of all lipids from LDL resulted in the complete loss of LAT activation. Removal of only neutral lipids by extraction with heptane retained up to 50% of the original activity, which was not increased further by reconstitution of the LDL with the extracted lipids. Hydrolysis of the diacylphosphoglycerides of the LDL with phospholipases resulted in complete loss of LAT activation which was partially restored by the addition of egg lecithin. Hydrolysis of more than 4% of LDL protein by trypsin led to a linear decrease in activity with complete loss of activity occurring when about 25% of the LDL protein is hydrolyzed. Modification of the arginine groups of LDL reversibly inhibited the activation of LAT. Modification of lysine residues of LDL by acetylation, acetoacetylation or succinylation also abolished its ability to activate lysolecithin acylation.

  8. The M Phase Kinase Greatwall (Gwl) Promotes Inactivation of PP2A/B55δ, a Phosphatase Directed Against CDK Phosphosites

    PubMed Central

    Castilho, Priscila V.; Williams, Byron C.; Mochida, Satoru; Zhao, Yong

    2009-01-01

    We have previously shown that Greatwall kinase (Gwl) is required for M phase entry and maintenance in Xenopus egg extracts. Here, we demonstrate that Gwl plays a crucial role in a novel biochemical pathway that inactivates, specifically during M phase, “antimitotic” phosphatases directed against phosphorylations catalyzed by cyclin-dependent kinases (CDKs). A major component of this phosphatase activity is heterotrimeric PP2A containing the B55δ regulatory subunit. Gwl is activated during M phase by Cdk1/cyclin B (MPF), but once activated, Gwl promotes PP2A/B55δ inhibition with no further requirement for MPF. In the absence of Gwl, PP2A/B55δ remains active even when MPF levels are high. The removal of PP2A/B55δ corrects the inability of Gwl-depleted extracts to enter M phase. These findings support the hypothesis that M phase requires not only high levels of MPF function, but also the suppression, through a Gwl-dependent mechanism, of phosphatase(s) that would otherwise remove MPF-driven phosphorylations. PMID:19793917

  9. Beneficial Effects of Simulated Gastro-Intestinal Digests of Fried Egg and Its Fractions on Blood Pressure, Plasma Lipids and Oxidative Stress in Spontaneously Hypertensive Rats

    PubMed Central

    Chakrabarti, Subhadeep; Morton, Jude S.; Panahi, Sareh; Kaufman, Susan; Davidge, Sandra T.; Wu, Jianping

    2014-01-01

    Abstract Background We have previously characterized several antihypertensive peptides in simulated digests of cooked eggs and showed blood pressure lowering property of fried whole egg digest. However, the long-term effects of this hydrolysate and its fractions on blood pressure are not known. Therefore, the objectives of the study were to determine the effects of long term administration of fried whole egg hydrolysate and its fractions (i.e. egg white and egg yolk) on regulation of blood pressure and associated factors in cardiovascular disease such as plasma lipid profile and tissue oxidative stress. Methods and Results We used spontaneously hypertensive rats (SHR), an animal model of essential hypertension. Hydrolysates of fried egg and its fractions were prepared by simulated gastro-intestinal digestion with pepsin and pancreatin. 16–17 week old male SHRs were orally administered fried whole egg hydrolysate, non-hydrolyzed fried whole egg, egg white hydrolysate or egg yolk hydrolysates (either defatted, or not) daily for 18 days. Blood pressure (BP) and heart rate were monitored by telemetry. Animals were sacrificed at the end of the treatment for vascular function studies and evaluating plasma lipid profile and tissue oxidative stress. BP was reduced by feeding fried whole egg hydrolysate but not by the non-hydrolyzed product suggesting a critical role for in vitro digestion in releasing anti-hypertensive peptides. Egg white hydrolysate and defatted egg yolk hydrolysate (but not egg yolk hydrolysate) also had similar effects. Reduction in BP was accompanied by the restoration of nitric oxide (NO) dependent vasorelaxation and reduction of plasma angiotensin II. Fried whole egg hydrolysate also reduced plasma levels of triglyceride although it was increased by the non-hydrolyzed sample. Additionally the hydrolyzed preparations attenuated tissue oxidative stress. Conclusion Our results demonstrate that fried egg hydrolysates exert anti-hypertensive effects, improve plasma lipid profile and attenuate tissue oxidative stress in vivo. PMID:25502445

  10. Naïve hosts of avian brood parasites accept foreign eggs, whereas older hosts fine-tune foreign egg discrimination during laying

    PubMed Central

    2014-01-01

    Background Many potential hosts of social parasites recognize and reject foreign intruders, and reduce or altogether escape the negative impacts of parasitism. The ontogenetic basis of whether and how avian hosts recognize their own and the brood parasitic eggs remains unclear. By repeatedly parasitizing the same hosts with a consistent parasitic egg type, and contrasting the responses of naïve and older breeders, we studied ontogenetic plasticity in the rejection of foreign eggs by the great reed warbler (Acrocephalus arundinaceus), a host species of the common cuckoo (Cuculus canorus). Results In response to experimental parasitism before the onset of laying, first time breeding hosts showed almost no egg ejection, compared to higher rates of ejection in older breeders. Young birds continued to accept foreign eggs when they were subjected to repeated parasitism, whereas older birds showed even higher ejection rates later in the same laying cycle. Conclusions Our results are consistent with the hypotheses that (i) naïve hosts need to see and learn the appearance of their own eggs to discriminate and reject foreign eggs, whereas (ii) experienced breeders possess a recognition template of their own eggs and reject parasitic eggs even without having to see their own eggs. However, we cannot exclude the possibility that other external cues and internal processes, accumulated simply with increasing age, may also modify age-specific patterns in egg rejection (e.g. more sightings of the cuckoo by older breeders). Future research should specifically track the potential role of learning in responses of individual hosts between first and subsequent breeding attempts by testing whether imprinting on a parasitized clutch reduces the rates of rejecting foreign eggs in subsequent parasitized clutches. PMID:25024736

  11. Antioxidant and ACE Inhibitory Bioactive Peptides Purified from Egg Yolk Proteins

    PubMed Central

    Yousr, Marwa; Howell, Nazlin

    2015-01-01

    Protein by-products from the extraction of lecithin from egg yolk can be converted into value-added products, such as bioactive hydrolysates and peptides that have potential health enhancing antioxidant, and antihypertensive properties. In this study, the antioxidant and angiotensin converting enzyme (ACE) inhibitory activities of peptides isolated and purified from egg yolk protein were investigated. Defatted egg yolk was hydrolyzed using pepsin and pancreatin and sequentially fractionated by ultrafiltration, followed by gel filtration to produce egg yolk gel filtration fractions (EYGF). Of these, two fractions, EYGF-23 and EYGF-33, effectively inhibited the peroxides and thiobarbituric acid reactive substance (TBARS) in an oxidizing linoleic acid model system. The antioxidant mechanism involved superoxide anion and hydroxyl radicals scavenging and ferrous chelation. The presence of hydrophobic amino acids such as tyrosine (Y) and tryptophan (W), in sequences identified by LC-MS as WYGPD (EYGF-23) and KLSDW (EYGF-33), contributed to the antioxidant activity and were not significantly different from the synthetic BHA antioxidant. A third fraction (EYGF-56) was also purified from egg yolk protein by gel filtration and exhibited high ACE inhibitory activity (69%) and IC50 value (3.35 mg/mL). The SDNRNQGY peptide (10 mg/mL) had ACE inhibitory activity, which was not significantly different from that of the positive control captopril (0.5 mg/mL). In addition, YPSPV in (EYGF-33) (10 mg/mL) had higher ACE inhibitory activity compared with captopril. These findings indicated a substantial potential for producing valuable peptides with antioxidant and ACE inhibitory activity from egg yolk. PMID:26690134

  12. Antioxidant and ACE Inhibitory Bioactive Peptides Purified from Egg Yolk Proteins.

    PubMed

    Yousr, Marwa; Howell, Nazlin

    2015-12-07

    Protein by-products from the extraction of lecithin from egg yolk can be converted into value-added products, such as bioactive hydrolysates and peptides that have potential health enhancing antioxidant, and antihypertensive properties. In this study, the antioxidant and angiotensin converting enzyme (ACE) inhibitory activities of peptides isolated and purified from egg yolk protein were investigated. Defatted egg yolk was hydrolyzed using pepsin and pancreatin and sequentially fractionated by ultrafiltration, followed by gel filtration to produce egg yolk gel filtration fractions (EYGF). Of these, two fractions, EYGF-23 and EYGF-33, effectively inhibited the peroxides and thiobarbituric acid reactive substance (TBARS) in an oxidizing linoleic acid model system. The antioxidant mechanism involved superoxide anion and hydroxyl radicals scavenging and ferrous chelation. The presence of hydrophobic amino acids such as tyrosine (Y) and tryptophan (W), in sequences identified by LC-MS as WYGPD (EYGF-23) and KLSDW (EYGF-33), contributed to the antioxidant activity and were not significantly different from the synthetic BHA antioxidant. A third fraction (EYGF-56) was also purified from egg yolk protein by gel filtration and exhibited high ACE inhibitory activity (69%) and IC50 value (3.35 mg/mL). The SDNRNQGY peptide (10 mg/mL) had ACE inhibitory activity, which was not significantly different from that of the positive control captopril (0.5 mg/mL). In addition, YPSPV in (EYGF-33) (10 mg/mL) had higher ACE inhibitory activity compared with captopril. These findings indicated a substantial potential for producing valuable peptides with antioxidant and ACE inhibitory activity from egg yolk.

  13. A Measure of the Auditory-perceptual Quality of Strain from Electroglottographic Analysis of Continuous Dysphonic Speech: Application to Adductor Spasmodic Dysphonia.

    PubMed

    Somanath, Keerthan; Mau, Ted

    2016-11-01

    (1) To develop an automated algorithm to analyze electroglottographic (EGG) signal in continuous dysphonic speech, and (2) to identify EGG waveform parameters that correlate with the auditory-perceptual quality of strain in the speech of patients with adductor spasmodic dysphonia (ADSD). Software development with application in a prospective controlled study. EGG was recorded from 12 normal speakers and 12 subjects with ADSD reading excerpts from the Rainbow Passage. Data were processed by a new algorithm developed with the specific goal of analyzing continuous dysphonic speech. The contact quotient, pulse width, a new parameter peak skew, and various contact closing slope quotient and contact opening slope quotient measures were extracted. EGG parameters were compared between normal and ADSD speech. Within the ADSD group, intra-subject comparison was also made between perceptually strained syllables and unstrained syllables. The opening slope quotient SO7525 distinguished strained syllables from unstrained syllables in continuous speech within individual subjects with ADSD. The standard deviations, but not the means, of contact quotient, EGGW50, peak skew, and SO7525 were different between normal and ADSD speakers. The strain-stress pattern in continuous speech can be visualized as color gradients based on the variation of EGG parameter values. EGG parameters may provide a within-subject measure of vocal strain and serve as a marker for treatment response. The addition of EGG to multidimensional assessment may lead to improved characterization of the voice disturbance in ADSD. Copyright © 2016 The Voice Foundation. Published by Elsevier Inc. All rights reserved.

  14. A measure of the auditory-perceptual quality of strain from electroglottographic analysis of continuous dysphonic speech: Application to adductor spasmodic dysphonia

    PubMed Central

    Somanath, Keerthan; Mau, Ted

    2016-01-01

    Objectives (1) To develop an automated algorithm to analyze electroglottographic (EGG) signal in continuous, dysphonic speech, and (2) to identify EGG waveform parameters that correlate with the auditory-perceptual quality of strain in the speech of patients with adductor spasmodic dysphonia (ADSD). Study Design Software development with application in a prospective controlled study. Methods EGG was recorded from 12 normal speakers and 12 subjects with ADSD reading excerpts from the Rainbow Passage. Data were processed by a new algorithm developed with the specific goal of analyzing continuous dysphonic speech. The contact quotient (CQ), pulse width (EGGW), a new parameter peak skew, and various contact closing slope quotient (SC) and contact opening slope quotient (SO) measures were extracted. EGG parameters were compared between normal and ADSD speech. Within the ADSD group, intra-subject comparison was also made between perceptually strained syllables and unstrained syllables. Results The opening slope quotient SO7525 distinguished strained syllables from unstrained syllables in continuous speech within individual ADSD subjects. The standard deviations, but not the means, of CQ, EGGW50, peak skew, and SO7525 were different between normal and ADSD speakers. The strain-stress pattern in continuous speech can be visualized as color gradients based on the variation of EGG parameter values. Conclusions EGG parameters may provide a within-subject measure of vocal strain and serve as a marker for treatment response. The addition of EGG to multi-dimensional assessment may lead to improved characterization of the voice disturbance in ADSD. PMID:26739857

  15. Larvicidal & ovicidal efficacy of Pithecellobium dulce (Roxb.) Benth. (Fabaceae) against Anopheles stephensi Liston & Aedes aegypti Linn. (Diptera: Culicidae)

    PubMed Central

    Govindarajan, M.; Rajeswary, M.; Sivakumar, R.

    2013-01-01

    Background & objectives: In view of the recently increased interest in developing plant origin insecticides as an alternative to chemical insecticide, this study was undertaken to assess the larvicidal and ovicidal potential of the crude hexane, benzene, chloroform, ethyl acetate and methanol solvent extracts from the medicinal plant Pithecellobium dulce against the mosquito vectors, Anopheles stephensi and Aedes aegypti (Diptera: Culicidae). Methods: Larvicidal activity of P. dulce plant extracts was studied in the range of 60 to 450 mg/l against early third instar larvae of An. stephensi and Ae. aegypti in the laboratory. The larval mortality was observed after 24 h of exposure. The ovicidal activity was determined against An. stephensi and Ae. aegypti to various concentrations ranging from 100 to 750 mg/l under the laboratory conditions. Mean per cent hatchability of the eggs were observed after 48 h post treatment. Results: All leaf and seed extracts showed moderate larvicidal and ovicidal effects; however, the highest larval mortality was found in methanol extract of leaf of P. dulce against the larvae of An. stephensi and Ae. aegypti with the LC50 and LC90 values 145.43, 155.78 mg/l and 251.23, 279.73 mg/l, respectively. The per cent hatchability was inversely proportional to the concentration of extract and directly proportional to the eggs. Zero hatchability was observed at 400 mg/l for leaf methanol extract and 625 mg/l for seed methanol extract of P. dulce against An. stephensi and Ae. aegypti, respectively. Compared to leaf extracts, seed extracts have low potency against the two mosquitoes. Interpretation & conclusions: The present results suggest that the leaf and seed extracts of P. dulce have the potential to be used as an ideal eco-friendly approach for the control of mosquitoes. PMID:24056567

  16. Biochemical Characterization and Vaccine Potential of a Heme-Binding Glutathione Transferase from the Adult Hookworm Ancylostoma caninum

    PubMed Central

    Zhan, Bin; Liu, Sen; Perally, Samirah; Xue, Jian; Fujiwara, Ricardo; Brophy, Peter; Xiao, Shuhua; Liu, Yueyuan; Feng, Jianjun; Williamson, Angela; Wang, Yan; Bueno, Lilian L.; Mendez, Susana; Goud, Gaddam; Bethony, Jeffrey M.; Hawdon, John M.; Loukas, Alex; Jones, Karen; Hotez, Peter J.

    2005-01-01

    We report the cloning and expression of Ac-GST-1, a novel glutathione S-transferase from the adult hookworm Ancylostoma caninum, and its possible role in parasite blood feeding and as a vaccine target. The predicted Ac-GST-1 open reading frame contains 207 amino acids (mass, 24 kDa) and exhibited up to 65% amino acid identity with other nematode GSTs. mRNA encoding Ac-GST-1 was detected in adults, eggs, and larval stages, but the protein was detected only in adult hookworm somatic extracts and excretory/secretory products. Using antiserum to the recombinant protein, Ac-GST-1 was immunolocalized to the parasite hypodermis and muscle tissue and weakly to the intestine. Recombinant Ac-GST-1 was enzymatically active, as determined by conjugation of glutathione to a model substrate, and exhibited a novel high-affinity binding site for hematin. The possible role of Ac-GST-1 in parasite heme detoxification during hemoglobin digestion or heme uptake prompted interest in evaluating it as a potential vaccine antigen. Vaccination of dogs with Ac-GST-1 resulted in a 39.4% reduction in the mean worm burden and 32.3% reduction in egg counts compared to control dogs following larval challenge, although the reductions were not statistically significant. However, hamsters vaccinated with Ac-GST-1 exhibited statistically significant worm reduction (53.7%) following challenge with heterologous Necator americanus larvae. These studies suggest that Ac-GST-1 is a possible drug and vaccine target for hookworm infection. PMID:16177370

  17. Role of atopy patch test for diagnosis of food allergy-related gastrointestinal symptoms in children.

    PubMed

    Boonyaviwat, Onsuree; Pacharn, Punchama; Jirapongsananuruk, Orathai; Vichyanond, Pakit; Visitsunthorn, Nualanong

    2015-12-01

    Double-blind, placebo-controlled food challenge is the gold standard for diagnosing food allergy. However, it is a time-consuming procedure and requires onsite medical supervision and resuscitating medicines and devices on hand. The objective of this study was to compare the atopy patch test (APT) with the oral food challenge test (OFC) in children with suspected food allergy-related gastrointestinal (GI) symptoms. A prospective self-controlled study enrolled children with a history of suspected food allergy-related GI symptoms. Skin prick test (SPT) and APT using lyophilized and commercial allergen extracts for cow's milk, egg, wheat, soy, and shrimp were evaluated, and OFC was performed. Thirty-nine patients (25 boys, median age 2.4 yrs) with 76 events of suspected food allergy-related GI symptoms were enrolled. SPT was positive in 11/76 events (14.5%). Sensitivity, specificity, predictive values, and likelihood ratio were calculated related to the food challenge outcome. Of 41 OFC, 30 (73.2%) were positive. APT using lyophilized allergen extracts yielded high sensitivity (80%) and high positive predictive value (85.7%). APT using commercial allergen extracts yielded low sensitivity (30%) but high specificity (90%). The negative predictive value of APT using lyophilized and commercial allergen extracts was 53.8% and 32.2%, respectively. All cases with positive APT using lyophilized allergen extracts together with positive SPT also had positive OFC. In contrast to commercial extracts, APT with lyophilized allergen extracts is reliable, safe, and maybe useful for the diagnosis of suspected food allergy-related GI symptoms in children. OFC is still needed in most of the cases. © 2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  18. The avian egg exhibits general allometric invariances in mechanical design.

    PubMed

    Juang, Jia-Yang; Chen, Pin-Yi; Yang, Da-Chang; Wu, Shang-Ping; Yen, An; Hsieh, Hsin-I

    2017-10-27

    The avian egg exhibits extraordinary diversity in size, shape and color, and has a key role in avian adaptive radiations. Despite extensive work, our understanding of the underlying principles that guide the "design" of the egg as a load-bearing structure remains incomplete, especially over broad taxonomic scales. Here we define a dimensionless number C, a function of egg weight, stiffness and dimensions, to quantify how stiff an egg is with respect to its weight after removing geometry-induced rigidity. We analyze eggs of 463 bird species in 36 orders across five orders of magnitude in body mass, and find that C number is nearly invariant for most species, including tiny hummingbirds and giant elephant birds. This invariance or "design guideline" dictates that evolutionary changes in shell thickness and Young's modulus, both contributing to shell stiffness, are constrained by changes in egg weight. Our analysis illuminates unique reproductive strategies of brood parasites, kiwis, and megapodes, and quantifies the loss of safety margin for contact incubation due to artificial selection and environmental toxins. Our approach provides a mechanistic framework for a better understanding of the mechanical design of the avian egg, and may provide clues to the evolutionary origin of contact incubation of amniote eggs.

  19. Cytoskeleton and gravity at work in the establishment of dorso-ventral polarity in the egg of Xenopus laevis

    NASA Astrophysics Data System (ADS)

    Ubbels, Geertje A.; Brom, Tim G.

    The establishment of polarities during early embryogenesis is essential for normal development. Amphibian eggs are appropriate models for studies on embryonic pattern formation. The animal-vegetal axis of the axially symmetrical amphibian egg originates during oogenesis and foreshadows the main body axis of the embryo. The dorso-ventral polarity is epigenetically established before first cleavage. Recent experiments strongly suggest that in the monospermic eggs of the anuran Xenopus laevis both the cytoskeleton and gravity act in the determination of the dorso-ventral polarity. In order to test the role of gravity in this process, eggs will be fertilized under microgravity conditions during the SL-D1 flight in 1985. In a fully automatic experiment container eggs will be kept under well-defined conditions and artificially fertilized as soon as microgravity is reached; eggs and embryos at different stages will then be fixed for later examination. Back on earth the material will be analysed and we will know whether fertilization under microgravity conditions is possible. If so, the relation of the dorso-ventral axis to the former sperm entry point will be determined on the whole embryos; in addition eggs and embryos will be analysed cytologically.

  20. Tracking living decapod larvae: mass staining of eggs with neutral red prior to hatching.

    PubMed

    Øresland, V; Horobin, R W

    2012-04-01

    Mass staining of decapod females carrying eggs, with subsequent identification of hatched larvae in the environment, is a research tool with great potential for field ecologists wishing to track the movements of larvae. For this to be achieved, however, numerous requirements must be met. These include adequate dye solubility, short staining time, dye penetration through different tissues, dye retention within the organism, absence of toxic and behavioral effects, low visibility to predators of stained larvae, no loss of staining owing to preservatives and low cost. The dye, neutral red, appears to meet most of these requirements. This dye was used in aliquots of 0.7 g/770 ml seawater applied to the females of Norway lobster (Nephrops norvegicus) and European lobster (Homarus gammarus) for 10 min. This procedure stained lobster eggs and embryos so that hatched larvae could be distinguished easily by fluorescence microscopy from larvae that hatched from unstained eggs. Stained larvae that were preserved in 4% formaldehyde in seawater were still stained after 1 year. Larvae should not come in contact with ethanol, because it extracts the dye rapidly.

  1. Mitotic trigger waves and the spatial coordination of the Xenopus cell cycle.

    PubMed

    Chang, Jeremy B; Ferrell, James E

    2013-08-29

    Despite the large size of the Xenopus laevis egg (approximately 1.2 mm diameter), a fertilized egg rapidly proceeds through mitosis in a spatially coordinated fashion. Mitosis is initiated by a bistable system of regulatory proteins centred on Cdk1 (refs 1, 2), raising the possibility that this spatial coordination could be achieved through trigger waves of Cdk1 activity. Using an extract system that performs cell cycles in vitro, here we show that mitosis does spread through Xenopus cytoplasm via trigger waves, propagating at a linear speed of approximately 60 µm min(-1). Perturbing the feedback loops that give rise to the bistability of Cdk1 changes the speed and dynamics of the waves. Time-lapse imaging of intact eggs argues that trigger waves of Cdk1 activation are responsible for surface contraction waves, ripples in the cell cortex that precede cytokinesis. These findings indicate that Cdk1 trigger waves help ensure the spatiotemporal coordination of mitosis in large eggs. Trigger waves may be an important general mechanism for coordinating biochemical events over large distances.

  2. Studies on some feed additives and materials giving partial protection against the suppressive effect of ochratoxin A on egg production of laying hens.

    PubMed

    Stoev, Stoycho D

    2010-06-01

    The protective effects of various feed supplements against the harmful effect of ochratoxin A on egg production and sexual maturation of two-weeks old Plymouth Rock female chicks designed for laying hens were studied. A significant protective effect of the feed additives or materials: water extract of artichoke (WEA), sesame seed (SS), Roxazyme-G (RG) and l-beta phenylalanine (PHE) against the suppressive effect of ochratoxin A (OTA) on egg production of laying hens was found. A similar protection was also seen on the toxic effect of OTA on various internal organs of the same hens. A significant protection was found against the decrease of the weight or the quantity of eggs as well as against the delay of the beginning of the laying period of chicks, both of which were provoked by ochratoxin A. These protective effects were strongest in chicks treated with SS or WEA, but were slightest in chicks treated with l-beta PHE. Copyright 2009 Elsevier Ltd. All rights reserved.

  3. Potential effect of the medicinal plants Calotropis procera, Ficus elastica and Zingiber officinale against Schistosoma mansoni in mice.

    PubMed

    Seif el-Din, Sayed H; El-Lakkany, Naglaa M; Mohamed, Mona A; Hamed, Manal M; Sterner, Olov; Botros, Sanaa S

    2014-02-01

    Calotropis procera (Ait.) R. Br. (Asclepiadaceae), Ficus elastica Roxb. (Moraceae) and Zingiber officinale Roscoe (Zingiberaceae) have been traditionally used to treat many diseases. The antischistosomal activity of these plant extracts was evaluated against Schistosoma mansoni. Male mice exposed to 80 ± 10 cercariae per mouse were divided into two batches. The first was divided into five groups: (I) infected untreated, while groups from (II-V) were treated orally (500 mg/kg for three consecutive days) by aqueous stem latex and flowers of C. procera, latex of F. elastica and ether extract of Z. officinale, respectively. The second batch was divided into four comparable groups (except Z. officinale-treated group) similarly treated as the first batch in addition to the antacid ranitidine (30 mg/kg) 1 h before extract administration. Safety, worm recovery, tissues egg load and oogram pattern were assessed. Calotropis procera latex and flower extracts are toxic (50-70% mortality) even in a small dose (250 mg/kg) before washing off their toxic rubber. Zingiber officinale extract insignificantly decrease (7.26%) S. mansoni worms. When toxic rubber was washed off and ranitidine was used, C. procera (stem latex and flowers) and F. elastica extracts revealed significant S. mansoni worm reductions by 45.31, 53.7 and 16.71%, respectively. Moreover, C. procera extracts produced significant reductions in tissue egg load (∼34-38.5%) and positively affected oogram pattern. The present study may be useful to supplement information with regard to C. procera and F. elastica antischistosomal activity and provide a basis for further experimental trials.

  4. Do females invest more into eggs when males sing more attractively? Postmating sexual selection strategies in a monogamous reed passerine.

    PubMed

    Krištofík, Ján; Darolová, Alžbeta; Majtan, Juraj; Okuliarová, Monika; Zeman, Michal; Hoi, Herbert

    2014-04-01

    Maternal investment can play an important role for offspring fitness, especially in birds, as females have to provide their eggs with all the necessary nutrients for the development of the embryo. It is known that this type of maternal investment can be influenced by the quality of the male partner. In this study, we first verify that male song is important in the mate choice of female Eurasian reed warblers, as males mate faster when their singing is more complex. Furthermore, female egg investment varies in relation to male song characteristics. Interestingly, clutch size, egg weight, or size, which can be considered as an high-cost investment, is not influenced by male song characteristics, whereas comparably low-cost investment types like investment into diverse egg components are adjusted to male song characteristics. In line with this, our results suggest that female allocation rules depend on investment type as well as song characteristics. For example, egg white lysozyme is positively correlated with male song complexity. In contrast, a negative correlation exists between-song speed and syllable repetitiveness and egg yolk weight as well as egg yolk testosterone concentration. Thus, our results suggest that female egg investment is related to male song performance in several aspects, but female investment patterns regarding various egg compounds are not simply correlated.

  5. Don't put all your eggs in one nest: spread them and cut time at risk.

    PubMed

    Andersson, Malte; Åhlund, Matti

    2012-09-01

    In many egg-laying animals, some females spread their clutch among several nests. The fitness effects of this reproductive tactic are obscure. Using mathematical modeling and field observations, we analyze an unexplored benefit of egg spreading in brood parasitic and other breeding systems: reduced time at risk for offspring. If a clutch takes many days to lay until incubation and embryo development starts after the last egg, by spreading her eggs a parasitic female can reduce offspring time in the vulnerable nest at risk of predation or other destruction. The model suggests that she can achieve much of this benefit by spreading her eggs among a few nests, even if her total clutch is large. Field data from goldeneye ducks Bucephala clangula show that egg spreading enables a fecund female to lay a clutch that is much larger than average without increasing offspring time at risk in a nest. This advantage increases with female condition (fecundity) and can markedly raise female reproductive success. These results help explain the puzzle of nesting parasites in some precocial birds, which lay eggs in the nests of other females before laying eggs in their own nest. Risk reduction by egg spreading may also play a role in the evolution of other breeding systems and taxa-for instance, polyandry with male parental care in some birds and fishes.

  6. Penguin chicks benefit from elevated yolk androgen levels under sibling competition.

    PubMed

    Poisbleau, Maud; Müller, Wendt; Carslake, David; Demongin, Laurent; Groothuis, Ton G G; Van Camp, Jeff; Eens, Marcel

    2012-01-01

    Crested penguins (genus Eudyptes) have a peculiar hatching pattern, with the first-laid egg (A-egg) hatching after the second-laid egg (B-egg) and chicks from A-eggs typically having a much lower survival probability. Maternal yolk androgens have been suggested to contribute to the competitive superiority of the B-chick in southern rockhopper penguins Eudyptes chrysocome, given their important role in mediating sibling competition in other species. We therefore increased the yolk androgen levels in freshly-laid eggs and examined the consequences for sibling competition--via effects on embryonic developmental times, chick growth and early survival. We placed one androgen-treated egg and one control egg into each foster nest, matching them for mass, laying date and laying order. The androgen treatment did not significantly affect embryonic developmental times or chick measurements at hatching. However, elevated yolk androgen levels benefitted chick growth in interaction with the number of siblings in a brood. Chicks from androgen-treated eggs had faster growth in the presence of a sibling than chicks from control eggs. Under these circumstances they also had a higher survival probability. Thus maternal androgens appear to reinforce the observed hatching pattern, facilitating brood reduction. This contrasts to most previous studies in other species where yolk androgens have been shown to compensate for the negative consequences of delayed hatching within the brood hierarchy.

  7. Compaction Kinetics on Single DNAs: Purified Nucleosome Reconstitution Systems versus Crude Extract

    PubMed Central

    Wagner, Gaudeline; Bancaud, Aurélien; Quivy, Jean-Pierre; Clapier, Cédric; Almouzni, Geneviève; Viovy, Jean-Louis

    2005-01-01

    Kinetics of compaction on single DNA molecules are studied by fluorescence videomicroscopy in the presence of 1), Xenopus egg extracts and 2), purified nucleosome reconstitution systems using a combination of histones with either the histone chaperone Nucleosome Assembly Protein (NAP-1) or negatively charged macromolecules such as polyglutamic acid and RNA. The comparison shows that the compaction rates can differ by a factor of up to 1000 for the same amount of histones, depending on the system used and on the presence of histone tails, which can be subjected to post-translational modifications. Reactions with purified reconstitution systems follow a slow and sequential mechanism, compatible with the deposition of one (H3-H4)2 tetramer followed by two (H2A-H2B) dimers. Addition of the histone chaperone NAP-1 increases both the rate of the reaction and the packing ratio of the final product. These stimulatory effects cannot be obtained with polyglutamic acid or RNA, suggesting that yNAP-1 impact on the reaction cannot simply be explained in terms of charge screening. Faster compaction kinetics and higher packing ratios are reproducibly reached with extracts, indicating a role of additional components present in this system. Data are discussed and models proposed to account for the kinetics obtained in our single-molecule assay. PMID:16100259

  8. Calcium and actin in the saga of awakening oocytes.

    PubMed

    Santella, Luigia; Limatola, Nunzia; Chun, Jong T

    2015-04-24

    The interaction of the spermatozoon with the egg at fertilization remains one of the most fascinating mysteries of life. Much of our scientific knowledge on fertilization comes from studies on sea urchin and starfish, which provide plenty of gametes. Large and transparent, these eggs have served as excellent model systems for studying egg activation and embryo development in seawater, a plain natural medium. Starfish oocytes allow the study of the cortical, cytoplasmic and nuclear changes during the meiotic maturation process, which can also be triggered in vitro by hormonal stimulation. These morphological and biochemical changes ensure successful fertilization of the eggs at the first metaphase. On the other hand, sea urchin eggs are fertilized after the completion of meiosis, and are particularly suitable for the study of sperm-egg interaction, early events of egg activation, and embryonic development, as a large number of mature eggs can be fertilized synchronously. Starfish and sea urchin eggs undergo abrupt changes in the cytoskeleton and ion fluxes in response to the fertilizing spermatozoon. The plasma membrane and cortex of an egg thus represent "excitable media" that quickly respond to the stimulus with the Ca(2+) swings and structural changes. In this article, we review some of the key findings on the rapid dynamic rearrangements of the actin cytoskeleton in the oocyte/egg cortex upon hormonal or sperm stimulation and their roles in the modulation of the Ca(2+) signals and in the control of monospermic fertilization. Copyright © 2015 Elsevier Inc. All rights reserved.

  9. Comparative proteome analysis of egg yolk plasma proteins during storage.

    PubMed

    Gao, Dan; Qiu, Ning; Liu, Yaping; Ma, Meihu

    2017-06-01

    Physical changes such as chicken egg white thinning and egg yolk flattening occur during storage, implying a decline in egg quality. To reveal the deteriorative process related to chicken egg internal quality, a comparative proteomic method was used in this study to analyze the alterations in egg yolk plasma proteins at different storage times (0, 20 and 40 days) under an ambient temperature of 22 ± 2 °C. Using two-dimensional electrophoresis followed by matrix-assisted laser desorption/ionization time-of-flight tandem mass spectrometry, 33 protein spots representing 12 proteins were identified with significant (P < 0.05) alterations in abundance at different storage times. The proteins that showed significant changes in abundance included serum albumin, vitellogenin fragments, IgY chains, ovalbumin, ovoinhibitor, α 2 -macroglobulin-like protein 1-like, hemopexin, transthyretin, apolipoprotein A-I and β 2 -glycoprotein I precursor. Accelerating degradation for most egg yolk plasma proteins was observed after prolonged storage (from day 20 to day 40). It is likely that the increased degradation of protease inhibitors such as ovoinhibitor and α 2 -macroglobulin-like protein 1-like during prolonged storage lead to an imbalance of protease and antiprotease in egg yolk, which may play a key role in the degradation of egg yolk proteins. These findings will provide an insight into the effects of storage on egg yolk protein changes and give a deeper understanding of the deteriorative process of chicken egg yolk. © 2016 Society of Chemical Industry. © 2016 Society of Chemical Industry.

  10. Agricultural restructuring and gastrointestinal parasitism in domestic ruminants on the rangelands of Kazakhstan.

    PubMed

    Morgan, E R; Torgerson, P R; Shaikenov, B S; Usenbayev, A E; Moore, A B M; Medley, G F; Milner-Gulland, E J

    2006-06-30

    In order to evaluate the epidemiology of parasitism in farmed ruminants in the changed agricultural context in Kazakhstan, 505 cattle, sheep and goats were surveyed for gastrointestinal parasitism using coprological examination, and a further thirty sheep and four goats using post mortem extraction of helminths. The dominant helminth genera were Marshallagia, Nematodirus and Trichostrongylus. Haemonchus was also present. Faecal egg counts were low (mean abundance 0-115 eggs per gram in different groups), and there was no relationship between faecal egg density and body condition score. Eggs of Nematodirus spp. were more common in sheep less than 1 yr of age, whereas those of the Trichostrongylidae were generally more common in adult sheep. Based on faecal egg counts, sheep grazing on common land close to permanent human settlements were generally no more heavily infected than those grazing the remote open plains. This could be due to a non-linear relationship between livestock numbers and stocking density. We suggest that cessation of strategic nematode control in the region following post-Soviet agricultural reform was offset by a general reduction in stocking density, such that parasite burdens remained low. However, recovery in livestock numbers is likely to lead to increased levels of infection and production loss unless sustainable control strategies are put in place.

  11. Impact of broiler egg storage on the relative expression of selected blastoderm genes associated with apoptosis, oxidative stress, and fatty acid metabolism.

    PubMed

    Bakst, M R; Welch, G R; Fetterer, R; Miska, K

    2016-06-01

    Cool temperature storage of eggs prior to incubation is a frequent practice by commercial broiler hatcheries. However, continued storage beyond 7 d leads to a progressive increase in the rate of early embryonic mortality. In this study, we examined the relative expression of 31 genes associated with fatty acid metabolism (8), apoptosis (7), and oxidative stress (16) pathways to better understand the basis of embryo mortality during egg storage. A total of 642 broiler eggs in 2 separate trials were subjected to the following egg treatments: stored 4 d (Control 1, C1); stored 21 d but subjected to short periods of incubation during egg storage (SPIDES); stored un-manipulated 21 d (NonSPIDES, NS); and stored 4 d then incubated for 10 h to advance the embryos to the same developmental stages as the SPIDES embryos (Control 2, C2). Hatchability trials (277 eggs) confirmed the efficacy of SPIDES compared to NS treatments in both trials. To determine relative expression of 31 selected genes, 365 blastoderms were isolated, staged, and flash frozen in batches of 5 to 10 blastoderms per vial (7 vials per egg treatment) prior to RNA extractions. Analysis of gene expression was performed using qRT-PCR and the results presented as relative expression normalized to C1. The relative expression of genes in which the SPIDES and C2 treatments were significantly up- or down-regulated in tandem indicated that the stage-specific expression of those genes was maintained by the SPIDES treatment. This study provides the relative gene expressions of blastodermal cells before and after prolonged egg storage as well as insight as to how SPIDES impacts blastodermal cell gene expression. Published by Oxford University Press on behalf of Poultry Science Association 2016. This work is written by (a) US Government employee(s) and is in the public domain in the US.

  12. Duplex quantitative real-time PCR assay for the detection and discrimination of the eggs of Toxocara canis and Toxocara cati (Nematoda, Ascaridoidea) in soil and fecal samples.

    PubMed

    Durant, Jean-Francois; Irenge, Leonid M; Fogt-Wyrwas, Renata; Dumont, Catherine; Doucet, Jean-Pierre; Mignon, Bernard; Losson, Bertrand; Gala, Jean-Luc

    2012-12-07

    Toxocarosis is a zoonotic disease caused by Toxocara canis (T. canis) and/or Toxocara cati (T. cati), two worldwide distributed roundworms which are parasites of canids and felids, respectively. Infections of humans occur through ingestion of embryonated eggs of T. canis or T. cati, when playing with soils contaminated with dogs or cats feces. Accordingly, the assessment of potential contamination of these areas with these roundworms eggs is paramount. A duplex quantitative real-time PCR (2qPCR) targeting the ribosomal RNA gene internal transcribed spacer (ITS2) has been developed and used for rapid and specific identification of T. canis and T. cati eggs in fecal and soil samples. The assay was set up on DNA samples extracted from 53 adult worms including T. canis, T. cati, T. leonina, Ascaris suum (A. suum) and Parascaris equorum (P. equorum). The assay was used to assess the presence of T. cati eggs in several samples, including 12 clean soil samples spiked with eggs of either T. cati or A. suum, 10 actual soil samples randomly collected from playgrounds in Brussels, and fecal samples from cats, dogs, and other animals. 2qPCR results on dogs and cats fecal samples were compared with results from microscopic examination. 2qPCR assay allowed specific detection of T. canis and T. cati, whether adult worms, eggs spiked in soil or fecal samples. The 2qPCR limit of detection (LOD) in spiked soil samples was 2 eggs per g of soil for a turnaround time of 3 hours. A perfect concordance was observed between 2qPCR assay and microscopic examination on dogs and cats feces. The newly developed 2qPCR assay can be useful for high throughput prospective or retrospective detection of T.canis and/or T. cati eggs in fecal samples as well as in soil samples from playgrounds, parks and sandpits.

  13. [Roles of sialic acids in sperm maturation and capacitation and sperm-egg recognition].

    PubMed

    Feng, Ying; Wang, Lin; Wu, Yi-Lun; Liu, Hong-Hua; Ma, Fang

    2016-10-01

    Sialic acids are a subset of nine-carbon alpha-keto aldonic acids involved in various biological functions. Sialic acid on the sperm surface is closely related to sperm maturation and capacitation and sperm-egg recognition, which makes sperm negatively charged to avoid accumulation and covers some antigenic determinants there to increase the survival rate of sperm in the female reproductive tract. The loss of sialic acids is an important factor mediating sperm capacitation. Moreover, the sialic acid at the extremity of the protein polymer is involved in signal identification in sperm-egg recognition. Here, we review the current understanding of sialic acids in sperm maturation and capacitation and sperm-egg recognition.

  14. A role for carbohydrate recognition in mammalian sperm-egg binding

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Clark, Gary F., E-mail: clarkgf@health.missouri.edu

    Highlights: • Mammalian sperm-egg binding as a carbohydrate dependent species recognition event. • The role of carbohydrate recognition in human, mouse and pig sperm-egg binding. • Historical perspective and future directions for research focused on gamete binding. - Abstract: Mammalian fertilization usually requires three sequential cell–cell interactions: (i) initial binding of sperm to the specialized extracellular matrix coating the egg known as the zona pellucida (ZP); (ii) binding of sperm to the ZP via the inner acrosomal membrane that is exposed following the induction of acrosomal exocytosis; and (iii) adhesion of acrosome-reacted sperm to the plasma membrane of the eggmore » cell, enabling subsequent fusion of these gametes. The focus of this review is on the initial binding of intact sperm to the mammalian ZP. Evidence collected over the past fifty years has confirmed that this interaction relies primarily on the recognition of carbohydrate sequences presented on the ZP by lectin-like egg binding proteins located on the plasma membrane of sperm. There is also evidence that the same carbohydrate sequences that mediate binding also function as ligands for lectins on lymphocytes that can inactivate immune responses, likely protecting the egg and the developing embryo up to the stage of blastocyst hatching. The literature related to initial sperm-ZP binding in the three major mammalian models (human, mouse and pig) is discussed. Historical perspectives and future directions for research related to this aspect of gamete adhesion are also presented.« less

  15. The protective role of endogenous bacterial communities in chironomid egg masses and larvae

    PubMed Central

    Senderovich, Yigal; Halpern, Malka

    2013-01-01

    Insects of the family Chironomidae, also known as chironomids, are distributed worldwide in a variety of water habitats. These insects display a wide range of tolerance toward metals and organic pollutions. Bacterial species known for their ability to degrade toxicants were identified from chironomid egg masses, leading to the hypothesis that bacteria may contribute to the survival of chironomids in polluted environments. To gain a better understanding of the bacterial communities that inhabit chironomids, the endogenous bacteria of egg masses and larvae were studied by 454-pyrosequencing. The microbial community of the egg masses was distinct from that of the larval stage, most likely due to the presence of one dominant bacterial Firmicutes taxon, which consisted of 28% of the total sequence reads from the larvae. This taxon may be an insect symbiont. The bacterial communities of both the egg masses and the larvae were found to include operational taxonomic units, which were closely related to species known as toxicant degraders. Furthermore, various bacterial species with the ability to detoxify metals were isolated from egg masses and larvae. Koch-like postulates were applied to demonstrate that chironomid endogenous bacterial species protect the insect from toxic heavy metals. We conclude that chironomids, which are considered pollution tolerant, are inhabited by stable endogenous bacterial communities that have a role in protecting their hosts from toxicants. This phenomenon, in which bacteria enable the continued existence of their host in hostile environments, may not be restricted only to chironomids. PMID:23804150

  16. Examining the relationships between egg cortisol and oxidative stress in developing wild sockeye salmon (Oncorhynchus nerka).

    PubMed

    Taylor, Jessica J; Sopinka, Natalie M; Wilson, Samantha M; Hinch, Scott G; Patterson, David A; Cooke, Steven J; Willmore, William G

    2016-10-01

    Maternally-derived hormones in oocytes, such as glucocorticoids (GCs), play a crucial role in embryo development in oviparous taxa. In fishes, maternal stressor exposure increases circulating and egg cortisol levels, the primary GC in fishes, as well as induces oxidative stress. Elevated egg cortisol levels modify offspring traits but whether maternal oxidative stress correlates with circulating and egg cortisol levels, and whether maternal/egg cortisol levels correlate with offspring oxidative stress have yet to be determined. The objective of this study was to examine the relationships among maternal and egg cortisol, and maternal and offspring oxidative stress to provide insight into the potential intergenerational effects of stressor exposure in sockeye salmon (Oncorhynchus nerka). Antioxidant concentration and oxidative stress were measured in maternal tissues (plasma, brain, heart and liver) as well as offspring developmental stages (pre-fertilization, 24h post-fertilization, eyed, and hatch), and were compared to both naturally-occurring and experimentally-elevated (via cortisol egg bath) levels of cortisol in eggs. Oxygen radical absorptive capacity of tissues from maternal sockeye salmon was measured spectrophotometrically and was not correlated with maternal or egg cortisol concentrations. Also, naturally-occurring and experimentally-elevated cortisol levels in eggs (to mimic maternal stress) did not affect oxidative stress or antioxidant capacity of the offspring. We conclude that the metrics of maternal stress examined in sockeye salmon (i.e., maternal/egg cortisol, maternal oxidative stress) are independent of each other, and that egg cortisol content does not influence offspring oxidative stress. Copyright © 2016 Elsevier Inc. All rights reserved.

  17. In search of genetic constraints limiting the evolution of egg size: direct and correlated responses to artificial selection on a prenatal maternal effector.

    PubMed

    Pick, J L; Hutter, P; Tschirren, B

    2016-06-01

    Maternal effects are an important force in nature, but the evolutionary dynamics of the traits that cause them are not well understood. Egg size is known to be a key mediator of prenatal maternal effects with an established genetic basis. In contrast to theoretical expectations for fitness-related traits, there is a large amount of additive genetic variation in egg size observed in natural populations. One possible mechanism for the maintenance of this variation is through genetic constraints caused by a shared genetic basis among traits. Here we created replicated, divergent selection lines for maternal egg investment in Japanese quail (Coturnix japonica) to quantify the role of genetic constraints in the evolution of egg size. We found that egg size responds rapidly to selection, accompanied by a strong response in all egg components. Initially, we observed a correlated response in body size, but this response declined over time, showing that egg size and body size can evolve independently. Furthermore, no correlated response in fecundity (measured as the proportion of days on which a female laid an egg) was observed. However, the response to selection was asymmetrical, with egg size plateauing after one generation of selection in the high but not the low investment lines. We attribute this pattern to the presence of genetic asymmetries, caused by directional dominance or unequal allele frequencies. Such asymmetries may contribute to the evolutionary stasis in egg size observed in natural populations, despite a positive association between egg size and fitness.

  18. In search of genetic constraints limiting the evolution of egg size: direct and correlated responses to artificial selection on a prenatal maternal effector

    PubMed Central

    Pick, J L; Hutter, P; Tschirren, B

    2016-01-01

    Maternal effects are an important force in nature, but the evolutionary dynamics of the traits that cause them are not well understood. Egg size is known to be a key mediator of prenatal maternal effects with an established genetic basis. In contrast to theoretical expectations for fitness-related traits, there is a large amount of additive genetic variation in egg size observed in natural populations. One possible mechanism for the maintenance of this variation is through genetic constraints caused by a shared genetic basis among traits. Here we created replicated, divergent selection lines for maternal egg investment in Japanese quail (Coturnix japonica) to quantify the role of genetic constraints in the evolution of egg size. We found that egg size responds rapidly to selection, accompanied by a strong response in all egg components. Initially, we observed a correlated response in body size, but this response declined over time, showing that egg size and body size can evolve independently. Furthermore, no correlated response in fecundity (measured as the proportion of days on which a female laid an egg) was observed. However, the response to selection was asymmetrical, with egg size plateauing after one generation of selection in the high but not the low investment lines. We attribute this pattern to the presence of genetic asymmetries, caused by directional dominance or unequal allele frequencies. Such asymmetries may contribute to the evolutionary stasis in egg size observed in natural populations, despite a positive association between egg size and fitness. PMID:26956564

  19. Involvement of 4E-BP phosphorylation in embryonic development of the silkworm, Bombyx mori.

    PubMed

    Gu, Shi-Hong; Young, Shun-Chieh; Tsai, Wen-Hsien; Lin, Ju-Ling; Lin, Pei-Ling

    2011-07-01

    Phosphorylation of the translational repressor 4E-binding protein (4E-BP) plays a critical role in regulating the overall translation levels in cells. In the present study, we investigated 4E-BP phosphorylation of Bombyx mori eggs by an immunoblot analysis of a conserved phospho-specific antibody to 4E-BP and demonstrated its role during embryonic development. When HCl treatment was applied to diapause-destined eggs at 20 h after oviposition, a dramatic increase in the phosphorylation of 4E-BP occurred 5 min after treatment with HCl, and high phosphorylation levels were maintained throughout embryonic stage in HCl-treated eggs compared to those in diapause (control) eggs. When HCl treatment was applied to diapause eggs on day 10 after oviposition, no dramatic activation in 4E-BP phosphorylation occurred, indicating stage-specific effects of HCl treatment. In both non-diapause eggs and eggs whose diapause had been terminated by chilling of diapausing eggs at 5°C for 70 days and then were transferred to 25°C, high phosphorylation levels of 4E-BP were also detected. Moreover, 4E-BP phosphorylation dramatically increased when dechorionated eggs were incubated in medium. The addition of rapamycin, a specific inhibitor of mammalian target of rapamycin (TOR) signaling, and LY294002, a phosphoinositide 3-kinase (PI3K) inhibitor, but not the mitogen-activated protein kinase (MAPK)/extracellular signal-regulated kinase (ERK) kinase (MEK) inhibitor, U0126, dose-dependently inhibited 4E-BP phosphorylation in dechorionated eggs, indicating that PI3K/TOR signaling is an upstream signaling event involved in 4E-BP phosphorylation. Examination of 4E-BP gene expression levels showed no differences between treatments with HCl and water in the first hour after treatment, indicating that changes in phosphorylation of 4E-BP upon HCl treatment are mainly regulated at the post-transcriptional level. In addition, MAPK pathways and glycogen synthase kinase (GSK)-3β phosphorylation were not significantly affected in the first hour after HCl treatment. These results demonstrate that the rapid phosphorylation of 4E-BP is an early signaling event in embryonic development in the eggs whose diapause initiation was prevented by HCl treatment, thus being involved in the embryonic development of B. mori. Copyright © 2011 Elsevier Ltd. All rights reserved.

  20. Discovery of a 240 million year old nematode parasite egg in a cynodont coprolite sheds light on the early origin of pinworms in vertebrates.

    PubMed

    Hugot, Jean-Pierre; Gardner, Scott L; Borba, Victor; Araujo, Priscilla; Leles, Daniela; Stock Da-Rosa, Átila Augusto; Dutra, Juliana; Ferreira, Luiz Fernando; Araújo, Adauto

    2014-11-13

    We report the discovery of a nematode parasite egg (Nemata: Oxyurida) from a coprolite closely associated with the remains of several species of Cynodontia, dated to 240 million years old. This finding is particularly significant because this is the oldest record of an oxyurid nematode yet discovered, and because the cynodonts are considered a stem-group of the mammals. We extracted material from a fully mineralized coprolite by both scraping the surface, and removing fragments from its interior with clean dental instruments used a single time. A single drop of glycerol from a new vial was added as a clearing reagent. Each slide was sealed with wax and examined with an optical microscope at 100× to 400× magnification. From one coprolite, 550 slides were examined; from 275 of these slides, sediment was examined that was scraped from the surface of the coprolite, and from the other 275 slides, material was examined that was extracted from the interior of the coprolite. All microscopic structures encountered were photographed, measured, and identified when possible. From the coprolite examined, we discovered an egg representing a new species of pinworm that, based on the egg structure, clearly places it in the family Heteroxynematidae. Nematodes of the order Oxyurida have very constrained life-histories, occurring only in animals that are not strictly carnivorous and also ingest large amounts of plant material. This fact enabled us to determine which species of cynodont, from several collected at the site in Brazil, are most likely the depositors of the coprolite, and therefore were the putative host of the parasite.

  1. Validation of a liquid chromatography-tandem mass spectrometry method for the simultaneous determination of sulfonamides, trimethoprim and dapsone in muscle, egg, milk and honey.

    PubMed

    Varenina, Ivana; Bilandžić, Nina; Kolanović, Božica Solomun; Božić, Đurđica; Sedak, Marija; Đokić, Maja; Varga, Ines

    2016-01-01

    A quantitative multi-residue method that includes 13 sulfonamides, trimethoprim and dapsone was developed and validated according to Commission Decision 2002/657/EC for muscle, milk egg and honey samples. For all matrices, the same extraction procedure was used. Samples were extracted with an acetone/dichloromethane mixture and cleaned up on aromatic sulfonic acid (SO3H) SPE cartridges. After elution and concentration steps, analytes were identified and quantified by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Data were acquired according to the multiple reaction-monitoring approach (MRM) and analytes were quantified both by the isotope dilution and the matrix-matched approaches calculating the response factors for the scanned product ions. The developed method shows good linearity, specificity, precision (repeatability and within-laboratory reproducibility), and trueness. Estimated CCβ for sulfonamides ranged between 5.6 and 8.2 µg kg(-1) for eggs, between 11.1 and 69.9 µg kg(-1) for milk, between 64.7 and 87.9 µg kg(-1) for muscle, and between 2.7 and 5.3 µg kg(-1) for honey. CCβ values for dapsone were 3.1, 0.6, 0.7 and 1.5 µg kg(-1) and for trimethoprim were 3.1, 6.7, 81.7 and 3.0 µg kg(-1) calculated for eggs, milk, muscle and honey, respectively. Recovery for all matrices was in the range from 89.1% and 109.7%. In matrix effect testing, no significant deviations were found between different samples of muscle and milk; however, a matrix effect was observed when testing different types of honey. The validation results demonstrate that the method is suitable for routine veterinary drug analysis and confirmation of suspect samples.

  2. Bioactive Egg Components and Inflammation

    PubMed Central

    Andersen, Catherine J.

    2015-01-01

    Inflammation is a normal acute response of the immune system to pathogens and tissue injury. However, chronic inflammation is known to play a significant role in the pathophysiology of numerous chronic diseases, such as cardiovascular disease, type 2 diabetes mellitus, and cancer. Thus, the impact of dietary factors on inflammation may provide key insight into mitigating chronic disease risk. Eggs are recognized as a functional food that contain a variety of bioactive compounds that can influence pro- and anti-inflammatory pathways. Interestingly, the effects of egg consumption on inflammation varies across different populations, including those that are classified as healthy, overweight, metabolic syndrome, and type 2 diabetic. The following review will discuss the pro- and anti-inflammatory properties of egg components, with a focus on egg phospholipids, cholesterol, the carotenoids lutein and zeaxanthin, and bioactive proteins. The effects of egg consumption of inflammation across human populations will additionally be presented. Together, these findings have implications for population-specific dietary recommendations and chronic disease risk. PMID:26389951

  3. Transmission of Enterobius vermicularis eggs through hands of school children in rural South Africa.

    PubMed

    Cranston, Imogen; Potgieter, Natasha; Mathebula, Sammy; Ensink, Jeroen H J

    2015-10-01

    Enterobius vermicularis is a helminth that is difficult to control, is found globally, especially in crowded conditions, and can be transmitted from person to person by contaminated hands. A newly developed method for the quantification of helminth eggs on hands was tested among schoolchildren in a rural South African region to look at the role hands play in helminth infection, and to determine the risk factors for hand contamination. The study found 16.6% of participants' hands positive for helminth eggs, with E. vermicularis most commonly identified. Egg concentrations on hands ranged from 0 to 57 eggs/2 hands. Gender, toilet type used at home, and not reporting to wash hands with soap before eating were all associated with the presence of eggs on hands. The study highlights the need to improve sanitation facilities, and promote handwashing with soap in schools to prevent transmission of E. vermicularis. Crown Copyright © 2015. Published by Elsevier B.V. All rights reserved.

  4. A nanostructural basis for gloss of avian eggshells

    PubMed Central

    Igic, Branislav; Fecheyr-Lippens, Daphne; Xiao, Ming; Chan, Andrew; Hanley, Daniel; Brennan, Patricia R. L.; Grim, Tomas; Waterhouse, Geoffrey I. N.; Hauber, Mark E.; Shawkey, Matthew D.

    2015-01-01

    The role of pigments in generating the colour and maculation of birds' eggs is well characterized, whereas the effects of the eggshell's nanostructure on the visual appearance of eggs are little studied. Here, we examined the nanostructural basis of glossiness of tinamou eggs. Tinamou eggs are well known for their glossy appearance, but the underlying mechanism responsible for this optical effect is unclear. Using experimental manipulations in conjunction with angle-resolved spectrophotometry, scanning electron microscopy, atomic force microscopy and chemical analyses, we show that the glossy appearance of tinamou eggshells is produced by an extremely smooth cuticle, composed of calcium carbonate, calcium phosphate and, potentially, organic compounds such as proteins and pigments. Optical calculations corroborate surface smoothness as the main factor producing gloss. Furthermore, we reveal the presence of weak iridescence on eggs of the great tinamou (Tinamus major), an optical effect never previously documented for bird eggs. These data highlight the need for further exploration into the nanostructural mechanisms for the production of colour and other optical effects of avian eggshells. PMID:25505139

  5. Evaluation of IgY Antibody as a Polyspecific Coombs-Reagent.

    PubMed

    Calzado, Esteban Justo Gutiérrez; Heredia, Marlene Toledano; Duharte, Jeorge Fernadez; Zarnani, Amir-Hassan

    2017-01-01

    During the last twenty years, the extraction of specific egg yolk (IgY) antibodies from the immunized chickens has been accepted as a useful alternative to the immunization of mammals. The aim of the present study was immunizing the chickens with Human Umbilical Cord Serum (HUCS) and the extraction of specific anti-human globulins (IgG, C3b, and C3d) antibodies from egg yolk in order to obtain polyspecific Coombs reagent. The novelty of this work was the achievement of a polyclonal reagent through a very cheap alternative method in accordance with all ethical regulations required for obtaining it. Three Leghorn hens (21 weeks old) were immunized four times for a period of 66 days with 20uL of HUCS mixed with PBS/FCA or FIA each time. The extraction of IgY antibodies was performed according to the method of lipid precipitation of yolk and using water soluble fraction as the reagent material. The resulting IgY antibody was characterized by SDS-PAGE and immunoelectrophoresis and tested for the presence of hetero-agglutinins by means of direct agglutination using human erythrocytes of all blood groups treated with 0.1% papain and for indirect Coombs-test to evaluate its specificity to fractions (C3b, C3d, C4d) of human complement and human IgG, respectively. Our findings show, that, the reagent obtained contains IgY and other 3 proteins (SDS-PAGE), and reacts specifically with plasma proteins, that migrate in β and ϒ regions. In immunoelectrophoresis, in addition, there is the presence of low hetero-agglutinins levels in IgY-preparation (3 lots), and the possibility to produce high amount (more than 500 ml/egg ) of polyspecific Coombs-reagent in chickens is also discussed. IgY-preparation (3 lots), and the possibility to produce high amount (more than 500 ml/egg ) of polyspecific Coombs-reagent in chickens with the originality to achieve a polyclonal reagent through a very cheap alternative method in accordance with all ethical regulations required for obtaining it, was also discussed.

  6. Evaluation of IgY Antibody as a Polyspecific Coombs-Reagent

    PubMed Central

    Calzado, Esteban Justo Gutiérrez; Heredia, Marlene Toledano; Duharte, Jeorge Fernadez; Zarnani, Amir-Hassan

    2017-01-01

    Background: During the last twenty years, the extraction of specific egg yolk (IgY) antibodies from the immunized chickens has been accepted as a useful alternative to the immunization of mammals. The aim of the present study was immunizing the chickens with Human Umbilical Cord Serum (HUCS) and the extraction of specific anti-human globulins (IgG, C3b, and C3d) antibodies from egg yolk in order to obtain polyspecific Coombs reagent. Methods: The novelty of this work was the achievement of a polyclonal reagent through a very cheap alternative method in accordance with all ethical regulations required for obtaining it. Three Leghorn hens (21 weeks old) were immunized four times for a period of 66 days with 20uL of HUCS mixed with PBS/FCA or FIA each time. The extraction of IgY antibodies was performed according to the method of lipid precipitation of yolk and using water soluble fraction as the reagent material. The resulting IgY antibody was characterized by SDS-PAGE and immunoelectrophoresis and tested for the presence of hetero-agglutinins by means of direct agglutination using human erythrocytes of all blood groups treated with 0.1% papain and for indirect Coombs-test to evaluate its specificity to fractions (C3b, C3d, C4d) of human complement and human IgG, respectively. Results: Our findings show, that, the reagent obtained contains IgY and other 3 proteins (SDS-PAGE), and reacts specifically with plasma proteins, that migrate in β and ϒ regions. In immunoelectrophoresis, in addition, there is the presence of low hetero-agglutinins levels in IgY-preparation (3 lots), and the possibility to produce high amount (more than 500 ml/egg) of polyspecific Coombs-reagent in chickens is also discussed. Conclusion: IgY-preparation (3 lots), and the possibility to produce high amount (more than 500 ml/egg) of polyspecific Coombs-reagent in chickens with the originality to achieve a polyclonal reagent through a very cheap alternative method in accordance with all ethical regulations required for obtaining it, was also discussed. PMID:28496948

  7. Consumer-phase Salmonella enterica serovar enteritidis risk assessment for egg-containing food products.

    PubMed

    Mokhtari, Amirhossein; Moore, Christina M; Yang, Hong; Jaykus, Lee-Ann; Morales, Roberta; Cates, Sheryl C; Cowen, Peter

    2006-06-01

    We describe a one-dimensional probabilistic model of the role of domestic food handling behaviors on salmonellosis risk associated with the consumption of eggs and egg-containing foods. Six categories of egg-containing foods were defined based on the amount of egg contained in the food, whether eggs are pooled, and the degree of cooking practiced by consumers. We used bootstrap simulation to quantify uncertainty in risk estimates due to sampling error, and sensitivity analysis to identify key sources of variability and uncertainty in the model. Because of typical model characteristics such as nonlinearity, interaction between inputs, thresholds, and saturation points, Sobol's method, a novel sensitivity analysis approach, was used to identify key sources of variability. Based on the mean probability of illness, examples of foods from the food categories ranked from most to least risk of illness were: (1) home-made salad dressings/ice cream; (2) fried eggs/boiled eggs; (3) omelettes; and (4) baked foods/breads. For food categories that may include uncooked eggs (e.g., home-made salad dressings/ice cream), consumer handling conditions such as storage time and temperature after food preparation were the key sources of variability. In contrast, for food categories associated with undercooked eggs (e.g., fried/soft-boiled eggs), the initial level of Salmonella contamination and the log10 reduction due to cooking were the key sources of variability. Important sources of uncertainty varied with both the risk percentile and the food category under consideration. This work adds to previous risk assessments focused on egg production and storage practices, and provides a science-based approach to inform consumer risk communications regarding safe egg handling practices.

  8. Role of specific IgE and skin-prick testing in predicting food challenge results to baked egg.

    PubMed

    Cortot, Catherine F; Sheehan, William J; Permaul, Perdita; Friedlander, James L; Baxi, Sachin N; Gaffin, Jonathan M; Dioun, Anahita F; Hoffman, Elaine B; Schneider, Lynda C; Phipatanakul, Wanda

    2012-01-01

    Previous studies suggest that children with egg allergy may be able to tolerate baked egg. Reliable predictors of a successful baked egg challenge are not well established. We examined egg white-specific IgE levels, skin-prick test (SPT) results, and age as predictors of baked egg oral food challenge (OFC) outcomes. We conducted a retrospective chart review of children, aged 2-18 years, receiving an egg white-specific IgE level, SPT, and OFC to baked egg from 2008 to 2010. Fifty-two oral baked egg challenges were conducted. Of the 52 challenges, 83% (n = 43) passed and 17% (n = 9) failed, including 2 having anaphylaxis. Median SPT wheal size was 12 mm (range, 0-35 mm) for passed challenges and 17 mm (range, 10-30 mm) for failed challenges (p = 0.091). The negative predictive value for passing the OFC was 100% (9 of 9) if SPT wheal size was <10 mm. Median egg white-specific IgE was 2.02 kU/L (range, <0.35-13.00 kU/L) for passed challenges and 1.52 kU/L (range, 0.51-6.10 kU/L) for failed challenges (p = 0.660). Receiver operating characteristic (ROC) curve analysis for SPT revealed an area under the curve (AUC) of 0.64. ROC curve analysis for egg white-specific IgE revealed an AUC of 0.63. There was no significant difference in age between patients who failed and those who passed (median = 8.8 years versus 7.0 years; p = 0.721). Based on our sample, SPT, egg white-specific IgE and age are not good predictors of passing a baked egg challenge. However, there was a trend for more predictability with SPT wheal size.

  9. Steroidogenic follicular response differences to LH and FSH treatment in low and high egg producing turkey hens

    USDA-ARS?s Scientific Manuscript database

    Low egg production in turkey hens is correlated with decreased ovulation frequency. Ovulation frequency is governed at the ovarian level by the steroid hormones progesterone and estradiol. The avian ovary is comprised of follicles in varying stages of maturation, which serve specialized roles in ste...

  10. M6 membrane protein plays an essential role in Drosophila oogenesis.

    PubMed

    Zappia, María Paula; Brocco, Marcela Adriana; Billi, Silvia C; Frasch, Alberto C; Ceriani, María Fernanda

    2011-01-01

    We had previously shown that the transmembrane glycoprotein M6a, a member of the proteolipid protein (PLP) family, regulates neurite/filopodium outgrowth, hence, M6a might be involved in neuronal remodeling and differentiation. In this work we focused on M6, the only PLP family member present in Drosophila, and ortholog to M6a. Unexpectedly, we found that decreased expression of M6 leads to female sterility. M6 is expressed in the membrane of the follicular epithelium in ovarioles throughout oogenesis. Phenotypes triggered by M6 downregulation in hypomorphic mutants included egg collapse and egg permeability, thus suggesting M6 involvement in eggshell biosynthesis. In addition, RNAi-mediated M6 knockdown targeted specifically to follicle cells induced an arrest of egg chamber development, revealing that M6 is essential in oogenesis. Interestingly, M6-associated phenotypes evidenced abnormal changes of the follicle cell shape and disrupted follicular epithelium in mid- and late-stage egg chambers. Therefore, we propose that M6 plays a role in follicular epithelium maintenance involving membrane cell remodeling during oogenesis in Drosophila.

  11. M6 Membrane Protein Plays an Essential Role in Drosophila Oogenesis

    PubMed Central

    Zappia, María Paula; Brocco, Marcela Adriana; Billi, Silvia C.; Frasch, Alberto C.; Ceriani, María Fernanda

    2011-01-01

    We had previously shown that the transmembrane glycoprotein M6a, a member of the proteolipid protein (PLP) family, regulates neurite/filopodium outgrowth, hence, M6a might be involved in neuronal remodeling and differentiation. In this work we focused on M6, the only PLP family member present in Drosophila, and ortholog to M6a. Unexpectedly, we found that decreased expression of M6 leads to female sterility. M6 is expressed in the membrane of the follicular epithelium in ovarioles throughout oogenesis. Phenotypes triggered by M6 downregulation in hypomorphic mutants included egg collapse and egg permeability, thus suggesting M6 involvement in eggshell biosynthesis. In addition, RNAi-mediated M6 knockdown targeted specifically to follicle cells induced an arrest of egg chamber development, revealing that M6 is essential in oogenesis. Interestingly, M6-associated phenotypes evidenced abnormal changes of the follicle cell shape and disrupted follicular epithelium in mid- and late-stage egg chambers. Therefore, we propose that M6 plays a role in follicular epithelium maintenance involving membrane cell remodeling during oogenesis in Drosophila. PMID:21603606

  12. Activity and immunodetection of lysozyme in earthworm Dendrobaena veneta (Annelida).

    PubMed

    Fiołka, Marta J; Zagaja, Mirosław P; Hułas-Stasiak, Monika; Wielbo, Jerzy

    2012-01-01

    In the present study, lysozyme-like activity against Micrococcus luteus was detected in the coelomic fluid, the extract from coelomocytes, intestine and in the homogenates from cocoons of Dendrobaena veneta. Four hours after immunization with Escherichia coli, the lysozyme activity in the coelomic fluid increased about three times and in the extract of coelomocytes - four times, in comparison to the control. In three cases: of the coelomic fluid, the homogenates from cocoons and the extract from coelomocytes, the antibody against HEWL (hen egg white lysozyme) recognized only one protein with a molecular mass of about 14.4 kDa. In the coelomic fluid, apart from the protein with molecular mass of 14.4 kDa the antibody directed against human lysozyme recognized an additional protein of 22 kDa. Using the bioautography technique after electrophoretic resolution of native proteins in acidic polyacrylamide gels, two lytic zones of M. luteus were observed in the case of the coelomic fluid and three after the analysis of the extract of coelomocytes and the egg homogenates. The results indicated the existence of several forms of lysozyme with a different electric charge in the analyzed D. veneta samples. The highest lysozyme activity in the intestine of D. veneta was observed in the midgut. The antibody directed against human lysozyme indicated a strong positive signal in epidermal and midgut cells of earthworm. Copyright © 2011 Elsevier Inc. All rights reserved.

  13. Multiclass screening method based on solvent extraction and liquid chromatography-tandem mass spectrometry for the determination of antimicrobials and mycotoxins in egg.

    PubMed

    Capriotti, Anna Laura; Cavaliere, Chiara; Piovesana, Susy; Samperi, Roberto; Laganà, Aldo

    2012-12-14

    A QuEChERS (Quick Easy Cheap Effective Rugged Safe)-like extraction method was developed for the simultaneous analysis of veterinary drugs and mycotoxins in hen eggs by liquid chromatography-tandem mass spectrometry (LC-MS/MS) with electrospray (ESI) source. Various classes of antimicrobials (tetracyclines, ionophores, coccidiostats, penicillins, cephalosporins, fluoroquinolones, sulfonamides) and mycotoxins (enniatins, beauvericin, ochratoxins, aflatoxins) were considered for the development of this method. Particular attention was devoted to extraction optimization: different solvents (acetone, acetonitrile and methanol), different pH values and different sample to extracting volume ratios were tested and evaluated in terms of recovery, relative standard deviation (RSD) and ESI signal suppression due to matrix effect. Chromatographic and mass spectrometric conditions were optimized to obtain the best instrumental performances for most of the analytes. Quantitative analysis was performed by means of matrix-matched calibration, in a range that varied depending on the analyte and its established maximum limit, when there was one. Recoveries at 100 μg kg(-1) spiking level were >62% (3

  14. Patterns and Drivers of Egg Pigment Intensity and Colour Diversity in the Ocean: A Meta-Analysis of Phylum Echinodermata.

    PubMed

    Montgomery, E M; Hamel, J-F; Mercier, A

    Egg pigmentation is proposed to serve numerous ecological, physiological, and adaptive functions in egg-laying animals. Despite the predominance and taxonomic diversity of egg layers, syntheses reviewing the putative functions and drivers of egg pigmentation have been relatively narrow in scope, centring almost exclusively on birds. Nonvertebrate and aquatic species are essentially overlooked, yet many of them produce maternally provisioned eggs in strikingly varied colours, from pale yellow to bright red or green. We explore the ways in which these colour patterns correlate with behavioural, morphological, geographic and phylogenetic variables in extant classes of Echinodermata, a phylum that has close phylogenetic ties with chordates and representatives in nearly all marine environments. Results of multivariate analyses show that intensely pigmented eggs are characteristic of pelagic or external development whereas pale eggs are commonly brooded internally. Of the five egg colours catalogued, orange and yellow are the most common. Yellow eggs are a primitive character, associated with all types of development (predominant in internal brooders), whereas green eggs are always pelagic, occur in the most derived orders of each class and are restricted to the Indo-Pacific Ocean. Orange eggs are geographically ubiquitous and may represent a 'universal' egg pigment that functions well under a diversity of environmental conditions. Finally, green occurs chiefly in the classes Holothuroidea and Ophiuroidea, orange in Asteroidea, yellow in Echinoidea, and brown in Holothuroidea. By examining an unprecedented combination of egg colours/intensities and reproductive strategies, this phylum-wide study sheds new light on the role and drivers of egg pigmentation, drawing parallels with theories developed from the study of more derived vertebrate taxa. The primary use of pigments (of any colour) to protect externally developing eggs from oxidative damage and predation is supported by the comparatively pale colour of equally large, internally brooded eggs. Secondarily, geographic location drives the evolution of egg colour diversity, presumably through the selection of better-adapted, more costly pigments in response to ecological pressure. © 2017 Elsevier Ltd. All rights reserved.

  15. Dynein-Based Accumulation of Membranes Regulates Nuclear Expansion in Xenopus laevis Egg Extracts.

    PubMed

    Hara, Yuki; Merten, Christoph A

    2015-06-08

    Nuclear size changes dynamically during development and has long been observed to correlate with the space surrounding the nucleus, as well as with the volume of the cell. Here we combine an in vitro cell-free system of Xenopus laevis egg extract with microfluidic devices to systematically analyze the effect of spatial constraints. The speed of nuclear expansion depended on the available space surrounding the nucleus up to a threshold volume in the nanoliter range, herein referred to as the nuclear domain. Under spatial constraints smaller than this nuclear domain, the size of microtubule-occupied space surrounding the nucleus turned out to be limiting for the accumulation of membranes around the nucleus via the motor protein dynein, therefore determining the speed of nuclear expansion. This mechanism explains how spatial information surrounding the nucleus, such as the positioning of the nucleus inside the cell, can control nuclear expansion. Copyright © 2015 Elsevier Inc. All rights reserved.

  16. Extracorporeal worm extraction of Diphyllobothrium nihonkaiense with amidotrizoic acid in a child.

    PubMed

    Shin, Hye Kyung; Roh, Joo-Hyung; Oh, Jae-Won; Ryu, Jae-Sook; Goo, Youn-Kyoung; Chung, Dong-Il; Kim, Yong Joo

    2014-12-01

    Infection cases of diphyllobothriid tapeworms are not much in the below teen-age group. We report a case of Diphyllobothrium nihonkaiense infection in a 13-year-old boy. He presented with severe fatigue, occasional abdominal pain at night time. He also had several episodes of tapeworm segment discharge in his stools. By his past history, he had frequently eaten raw fish including salmon and trout with his families. Numerous eggs of diphyllobothriid tapeworm were detected in the fecal examination. We introduced amidotrizoic acid as a cathartic agent through nasogastroduodenal tube and let nearly whole length (4.75 m) of D. nihonkaiense be excreted through his anus. After a single dose of praziquantel, the child's stool showed no further eggs, and his symptoms disappeared. The evacuated worm was identified as D. nihonkaiense by mitochondrial cox1 gene analysis. Here we report a successful extracorporeal worm extraction from an infection case of D. nihonkaiense by the injection of amidotrizoic acid.

  17. Extracorporeal Worm Extraction of Diphyllobothrium nihonkaiense with Amidotrizoic Acid in a Child

    PubMed Central

    Shin, Hye Kyung; Roh, Joo-Hyung; Oh, Jae-Won; Ryu, Jae-Sook; Goo, Youn-Kyoung; Chung, Dong-Il

    2014-01-01

    Infection cases of diphyllobothriid tapeworms are not much in the below teen-age group. We report a case of Diphyllobothrium nihonkaiense infection in a 13-year-old boy. He presented with severe fatigue, occasional abdominal pain at night time. He also had several episodes of tapeworm segment discharge in his stools. By his past history, he had frequently eaten raw fish including salmon and trout with his families. Numerous eggs of diphyllobothriid tapeworm were detected in the fecal examination. We introduced amidotrizoic acid as a cathartic agent through nasogastroduodenal tube and let nearly whole length (4.75 m) of D. nihonkaiense be excreted through his anus. After a single dose of praziquantel, the child's stool showed no further eggs, and his symptoms disappeared. The evacuated worm was identified as D. nihonkaiense by mitochondrial cox1 gene analysis. Here we report a successful extracorporeal worm extraction from an infection case of D. nihonkaiense by the injection of amidotrizoic acid. PMID:25548421

  18. Autocatalytic microtubule nucleation determines the size and mass of Xenopus laevis egg extract spindles.

    PubMed

    Decker, Franziska; Oriola, David; Dalton, Benjamin; Brugués, Jan

    2018-01-11

    Regulation of size and growth is a fundamental problem in biology. A prominent example is the formation of the mitotic spindle, where protein concentration gradients around chromosomes are thought to regulate spindle growth by controlling microtubule nucleation. Previous evidence suggests that microtubules nucleate throughout the spindle structure. However, the mechanisms underlying microtubule nucleation and its spatial regulation are still unclear. Here, we developed an assay based on laser ablation to directly probe microtubule nucleation events in Xenopus laevis egg extracts. Combining this method with theory and quantitative microscopy, we show that the size of a spindle is controlled by autocatalytic growth of microtubules, driven by microtubule-stimulated microtubule nucleation. The autocatalytic activity of this nucleation system is spatially regulated by the limiting amounts of active microtubule nucleators, which decrease with distance from the chromosomes. This mechanism provides an upper limit to spindle size even when resources are not limiting. © 2018, Decker et al.

  19. Nest materials as a source of genetic data for avian ecological studies

    USGS Publications Warehouse

    Pearce, J.M.; Fields, R.L.; Scribner, K.T.

    1997-01-01

    We examined the utility of feathers and egg shell membranes, deposited in the nests of Spectacled Eiders (Somateria fischeri), as a source of DNA for genetic studies at both the population and individual level. The potential for feather DNA contamination as a result of female behavioral interactions (e.g. nest parasitism), reuse of nest sites from previous years, or other unknown occurrences was acknowledged and specifically tested. DNA was successfully extracted from both feathers and egg shell membranes and waterfowl microsatellite loci were used to construct individual genotypes. We found no difference in the genotypes obtained from nest feathers or blood of the incubating female. Detection of nest feather contamination was possible with as little as one feather when samples from multiple females were intentionally mixed. Triplicate DNA extractions from 33 nests provided a means of detecting contamination in 3 nests. Egg membranes proved a viable source of offspring DNA and can contribute valuable data to investigations of parentage when assayed jointly with maternal feather DNA. Nest materials provide an efficient, non-invasive method of genetic sampling that can be readily incorporated into field research. However, the natural history traits and mating strategies of a species must be considered during sample collection to identify the possible sources of nest materials (e.g., paternal, maternal, parasite, etc.). Specific experiments should also be designed to test sampling assumptions.

  20. Determination of malachite green residues in the eggs, fry, and adult muscle-tissue of rainbow-trout (Oncorhynchus-mykiss)

    USGS Publications Warehouse

    Allen, John L.; Gofus, J.E.; Meinertz, Jeffery R.

    1994-01-01

    Malachite green, an effective antifungal therapeutant used in fish culture, is a known teratogen. We developed a method to simultaneously detect both the chromatic and leuco forms of malachite green residues in the eggs, fry, and adult muscle tissue of rainbow trout (oncorhynchus mykiss). Homogenates of these tissues were fortified with [c-14] malachite green chloride and extracted with 1% (v/v) acetic acid in acetonitrile or in methanol. The extracts were partitioned with chloroform, dried, redissolved in mobile phase, and analyzed by liquid chromatography (lc) with postcolumn oxidation of leuco malachite green to the chromatic form. Lc fractions were collected every 30 s for quantitation by scintillation counting. Recoveries of total [c-14] malachite green chloride residue were 85 and 98% in eggs fortified with labeled malachite green at concentrations of 0.5 And 1.00 Mug/g, respectively; 68% in fry similarly fortified at a concentration of 0.65 Mug/g; and 66% in muscle homogenate similarly fortified at a level of 1.00 Mug/g. The method was tested under operational conditions by exposing adult rainbow trout to 1.00 Mg/l [c-14] malachite green chloride bath for 1 h. Muscle samples analyzed by sample oxidation and scintillation counting contained 1.3 And 0.5 Mug/g total malachite green chloride residues immediately after exposure and after a 5-day withdrawal period, respectively.

  1. Reconstruction of gastric slow wave from finger photoplethysmographic signal using radial basis function neural network.

    PubMed

    Mohamed Yacin, S; Srinivasa Chakravarthy, V; Manivannan, M

    2011-11-01

    Extraction of extra-cardiac information from photoplethysmography (PPG) signal is a challenging research problem with significant clinical applications. In this study, radial basis function neural network (RBFNN) is used to reconstruct the gastric myoelectric activity (GMA) slow wave from finger PPG signal. Finger PPG and GMA (measured using Electrogastrogram, EGG) signals were acquired simultaneously at the sampling rate of 100 Hz from ten healthy subjects. Discrete wavelet transform (DWT) was used to extract slow wave (0-0.1953 Hz) component from the finger PPG signal; this slow wave PPG was used to reconstruct EGG. A RBFNN is trained on signals obtained from six subjects in both fasting and postprandial conditions. The trained network is tested on data obtained from the remaining four subjects. In the earlier study, we have shown the presence of GMA information in finger PPG signal using DWT and cross-correlation method. In this study, we explicitly reconstruct gastric slow wave from finger PPG signal by the proposed RBFNN-based method. It was found that the network-reconstructed slow wave provided significantly higher (P < 0.0001) correlation (≥ 0.9) with the subject's EGG slow wave than the correlation obtained (≈0.7) between the PPG slow wave from DWT and the EEG slow wave. Our results showed that a simple finger PPG signal can be used to reconstruct gastric slow wave using RBFNN method.

  2. Transfer of Maternal Antibodies against Avian Influenza Virus in Mallards (Anas platyrhynchos)

    PubMed Central

    van Dijk, Jacintha G. B.; Mateman, A. Christa; Klaassen, Marcel

    2014-01-01

    Maternal antibodies protect chicks from infection with pathogens early in life and may impact pathogen dynamics due to the alteration of the proportion of susceptible individuals in a population. We investigated the transfer of maternal antibodies against avian influenza virus (AIV) in a key AIV host species, the mallard (Anas platyrhynchos). Combining observations in both the field and in mallards kept in captivity, we connected maternal AIV antibody concentrations in eggs to (i) female body condition, (ii) female AIV antibody concentration, (iii) egg laying order, (iv) egg size and (v) embryo sex. We applied maternity analysis to the eggs collected in the field to account for intraspecific nest parasitism, which is reportedly high in Anseriformes, detecting parasitic eggs in one out of eight clutches. AIV antibody prevalence in free-living and captive females was respectively 48% and 56%, with 43% and 24% of the eggs receiving these antibodies maternally. In both field and captive study, maternal AIV antibody concentrations in egg yolk correlated positively with circulating AIV antibody concentrations in females. In the captive study, yolk AIV antibody concentrations correlated positively with egg laying order. Female body mass and egg size from the field and captive study, and embryos sex from the field study were not associated with maternal AIV antibody concentrations in eggs. Our study indicates that maternal AIV antibody transfer may potentially play an important role in shaping AIV infection dynamics in mallards. PMID:25386907

  3. Demographics and practices of semi-intensive free-range farming systems in Australia with an outdoor stocking density of ≤1500 hens/hectare

    PubMed Central

    Ruhnke, Isabelle; de Koning, Carolyn; Drake, Kelly; Skerman, Alan G.; Hinch, Geoff N.; Glatz, Philip C.

    2017-01-01

    Baseline information on demographics and practices on semi-intensive free-range egg farms with an outdoor stocking density of ≤1500 hens/hectare in Australia is presented. Free-range egg production is changing the structure of the egg industry in Australia and a broad variety and tiers of free-range systems have emerged due to lack of concrete legislative standards on outdoor stocking densities in the past. Information was extracted from a pre-existing online free-range poultry survey dataset, consisting of a total of 79 questions related to nutrition, pasture management, welfare and health, animal housing, environmental impact and economics. Forty-one free-range egg farms, with an outdoor stocking density of ≤1500 hens/hectare, were identified in the dataset from all major Australian states. Two types of semi-intensive free-range housing systems were documented: mobile (modified caravan/trailer) housing (56%), and fixed sheds (44%). Seventy-two percent of respondents reported >75% of the hens in the flock used the outdoor range. All respondents reported ingestion of range components by hens in the form of vegetation, insects, stones and grit. Up to 10% mortality was reported by 40% respondents with predation (34%), cannibalism (29%), heat stress (24%) and grass impaction (19.5%) as major causes. Biosecurity on farms was sub-optimal with 8 of the 10 actions implemented by <50% respondents. Customer demand, consumer sentiment and welfare were the major factors for farmers moving into free-range egg production. This study resulted in identification of current practices and key challenges on semi-intensive free-range egg farms. Applied research and communication of results to farmers is highly recommended to ensure optimum health and welfare of free-range laying hens and sustained egg production. PMID:29065169

  4. Chilling requirements for hatching of a New Zealand isolate of Nematodirus filicollis.

    PubMed

    Oliver, A-M B; Pomroy, W E; Ganesh, S; Leathwick, D M

    2016-08-15

    The eggs of some species of the parasitic nematode Nematodirus require a period of chilling before they can hatch; N. filicollis is one such species. This study investigated this requirement for chilling in a New Zealand strain of this species. Eggs of N. filicollis were extracted from lamb's faeces and incubated at 20°C to allow development to the third stage larvae within the egg. These eggs were then placed into tissue culture plates and incubated at: 2.7°C (±0.99), 3.6°C (±0.90), 4.7°C (±0.35), 6.4°C (±0.37), 8.0°C (±1.54) or 9.9°C (±0.14) for up to 224 days. At 14day intervals until day 84, then every 28 days, one plate was removed from each temperature and placed at 13.1°C (±0.44) for 14 days. Eggs were then assessed for hatching. From this data, chill units were calculated by subtracting the culture temperature from a constant threshold of 11°C and multiplying by the number of days for which the sample was cultured; then the Gompertz model fitted. Even though hatching overall was low, a greater proportion of eggs hatched with chill accumulation. Maximum hatching of eggs required 800-1000 chill units. Consequently in the field, more than one season of chilling would be required before hatching. As such a generation time could take more than one year to complete. This is different to the hatching dynamics of N. spathiger, the other main species found in New Zealand sheep, which does not display this requirement for chilling and hatches immediately once the third stage larvae are developed. Copyright © 2016 Elsevier B.V. All rights reserved.

  5. Demographics and practices of semi-intensive free-range farming systems in Australia with an outdoor stocking density of ≤1500 hens/hectare.

    PubMed

    Singh, Mini; Ruhnke, Isabelle; de Koning, Carolyn; Drake, Kelly; Skerman, Alan G; Hinch, Geoff N; Glatz, Philip C

    2017-01-01

    Baseline information on demographics and practices on semi-intensive free-range egg farms with an outdoor stocking density of ≤1500 hens/hectare in Australia is presented. Free-range egg production is changing the structure of the egg industry in Australia and a broad variety and tiers of free-range systems have emerged due to lack of concrete legislative standards on outdoor stocking densities in the past. Information was extracted from a pre-existing online free-range poultry survey dataset, consisting of a total of 79 questions related to nutrition, pasture management, welfare and health, animal housing, environmental impact and economics. Forty-one free-range egg farms, with an outdoor stocking density of ≤1500 hens/hectare, were identified in the dataset from all major Australian states. Two types of semi-intensive free-range housing systems were documented: mobile (modified caravan/trailer) housing (56%), and fixed sheds (44%). Seventy-two percent of respondents reported >75% of the hens in the flock used the outdoor range. All respondents reported ingestion of range components by hens in the form of vegetation, insects, stones and grit. Up to 10% mortality was reported by 40% respondents with predation (34%), cannibalism (29%), heat stress (24%) and grass impaction (19.5%) as major causes. Biosecurity on farms was sub-optimal with 8 of the 10 actions implemented by <50% respondents. Customer demand, consumer sentiment and welfare were the major factors for farmers moving into free-range egg production. This study resulted in identification of current practices and key challenges on semi-intensive free-range egg farms. Applied research and communication of results to farmers is highly recommended to ensure optimum health and welfare of free-range laying hens and sustained egg production.

  6. Bacterial Community Composition Associated with Chironomid Egg Masses

    PubMed Central

    Senderovich, Yigal; Halpern, Malka

    2012-01-01

    Chironomids (Diptera: Chironomidae) are the most widely distributed and often the most abundant insect in freshwater. They undergo a complete metamorphosis of four life stages, of which the egg, larva, and pupae are aquatic and the adult is terrestrial. Chironomid egg masses were found to be natural reservoirs of Vibrio cholerae and Aeromonas species. To expand the knowledge of the endogenous bacterial community associated with chironomid egg masses, denaturing gradient gel electrophoresis and clone analysis of 16S rRNA gene libraries were used in this study. Bacterial community composition associated with chironomid egg masses was found to be stable among different sampling periods. Cloned libraries of egg masses revealed that about 40% of the clones were related to bacteria known to degrade various toxicants. These findings were further supported when bacterial species that showed resistance to different toxic metals were isolated from egg masses and larval samples. Chironomids are found under a wide range of water conditions and are able to survive pollutants. However, little is known about their protective mechanisms under these conditions. Chironomid egg masses are inhabited by a stable endogenous bacterial community, which may potentially play a role in protecting chironomids from toxicants in polluted environments. Further study is needed to support this hypothesis. PMID:23461272

  7. Increase in membrane thickness during development compensates for eggshell thinning due to calcium uptake by the embryo in falcons

    NASA Astrophysics Data System (ADS)

    Castilla, Aurora M.; van Dongen, Stefan; Herrel, Anthony; Francesch, Amadeu; Martínez de Aragón, Juan; Malone, Jim; José Negro, Juan

    2010-02-01

    We compared membrane thickness of fully developed eggs with those of non-developed eggs in different endangered falcon taxa. To our knowledge, membrane thickness variation during development has never been examined before in falcons or any other wild bird. Yet, the egg membrane constitutes an important protective barrier for the developing embryo. Because eggshell thinning is a general process that occurs during bird development, caused by calcium uptake by the embryo, eggs are expected to be less protected and vulnerable to breakage near the end of development. Thus, egg membranes could play an important protective role in the later stages of development by getting relatively thicker. We used linear mixed models to explore the variation in membrane thickness ( n = 378 eggs) in relation to developmental stage, taxon, female age, mass and identity (73 females), egg-laying sequence (105 clutches) and the study zone. Our results are consistent with the prediction that egg membranes are thicker in fully developed eggs than in non-developed eggs, suggesting that the increase in membrane thickness during development may compensate for eggshell thinning. In addition, our data shown that thicker membranes are associated with larger, heavier and relatively wider eggs, as well as with eggs that had thinner eggshells. Egg-laying sequence, female age and the study zone did not explain the observed variation of membrane thickness in the falcon taxa studied. As we provide quantitative data on membrane thickness variation during development in falcons not subjected to contamination or food limitation (i.e. bred under captive conditions), our data may be used as a reference for studies on eggs from natural populations. Considering the large variation in membrane thickness and the multiple factors affecting on it and its importance in the protection of the embryo, we encourage other researchers to include measurements on membranes in studies exploring eggshell thickness variation.

  8. Changes in glutathione redox cycle during diapause determination and termination in the bivoltine silkworm, Bombyx mori.

    PubMed

    Zhao, Lin-Chuan; Hou, Yi-Sheng; Sima, Yang-Hu

    2014-02-01

    To explore whether glutathione regulates diapause determination and termination in the bivoltine silkworm Bombyx mori, we monitored the changes in glutathione redox cycle in the ovary of both diapause- and nondiapause-egg producers, as well as those in diapause eggs incubated at different temperatures. The activity of thioredoxin reductase (TrxR) was detected in ovaries but not in eggs, while neither ovaries nor eggs showed activity of glutathione peroxidase. A lower reduced glutathione/oxidized glutathione (GSH/GSSG) ratio was observed in the ovary of diapause-egg producers, due to weaker reduction of oxidized glutathione (GSSG) to the reduced glutathione (GSH) catalyzed by glutathione reductase (GR) and TrxR. This indicates an oxidative shift in the glutathione redox cycle during diapause determination. Compared with the 25°C-treated diapause eggs, the 5°C-treated diapause eggs showed lower GSH/GSSG ratio, a result of stronger oxidation of GSH catalyzed by thioredoxin peroxidase and weaker reduction of GSSG catalyzed by GR. Our study demonstrated the important regulatory role of glutathione in diapause determination and termination of the bivoltine silkworm. © 2013 Institute of Zoology, Chinese Academy of Sciences.

  9. Bacteria associated with Amblyomma cajennense tick eggs

    PubMed Central

    Machado-Ferreira, Erik; Vizzoni, Vinicius Figueiredo; Piesman, Joseph; Gazeta, Gilberto Salles; Soares, Carlos Augusto Gomes

    2015-01-01

    Abstract Ticks represent a large group of pathogen vectors that blood feed on a diversity of hosts. In the Americas, the Ixodidae ticks Amblyomma cajennense are responsible for severe impact on livestock and public health. In the present work, we present the isolation and molecular identification of a group of culturable bacteria associated with A. cajennense eggs from females sampled in distinct geographical sites in southeastern Brazil. Additional comparative analysis of the culturable bacteria from Anocentor nitens, Rhipicephalus sanguineus and Ixodes scapularis tick eggs were also performed. 16S rRNA gene sequence analyses identified 17 different bacterial types identified as Serratia marcescens, Stenotrophomonas maltophilia, Pseudomonas fluorescens, Enterobacter spp., Micrococcus luteus, Ochrobactrum anthropi, Bacillus cereus and Staphylococcus spp., distributed in 12 phylogroups. Staphylococcus spp., especially S. sciuri, was the most prevalent bacteria associated with A. cajennense eggs, occurring in 65% of the samples and also frequently observed infecting A. nitens eggs. S. maltophilia, S. marcescens and B. cereus occurred infecting eggs derived from specific sampling sites, but in all cases rising almost as pure cultures from infected A. cajennense eggs. The potential role of these bacterial associations is discussed and they possibly represent new targets for biological control strategies of ticks and tick borne diseases. PMID:26537602

  10. O-polysaccharide is important for Salmonella Pullorum survival in egg albumen, and virulence and colonization in chicken embryos.

    PubMed

    Guo, Rongxian; Li, Zhuoyang; Jiao, Yang; Geng, Shizhong; Pan, Zhiming; Chen, Xiang; Li, Qiuchun; Jiao, Xinan

    2017-10-01

    The pathogen Salmonella Pullorum is the causative agent of persistent systemic infection of poultry, leading to economic losses in developing countries due to morbidity, mortality and reduction in egg production. These infections may result in vertical transmission to eggs or progeny. Limited information is available regarding the mechanisms involved in the survival of Salmonella Pullorum in egg albumen and developing chicken embryos. Hence, we investigated the role of O-polysaccharide in the contamination of eggs and the colonization of chicken embryos. Compared with the wild-type strain, the isogenic waaL mutant exhibited an O-antigen-deficient rough phenotype, and increased sensitivity to egg albumen and chicken serum, as well as reduced adherence to DF-1 cells. Infection with Salmonella Pullorum lacking O-polysaccharide resulted in significantly reduced embryo lethality and bacterial colonization. These results suggest that O-polysaccharide is essential for Salmonella Pullorum colonization in eggs, both post-lay and developing embryos. The chicken embryo infection model could be used to characterize the interaction between Salmonella Pullorum and developing embryos, and it will also contribute to the development of more rational vaccines to protect laying hens and embryos.

  11. The extraembryonic serosa protects the insect egg against desiccation

    PubMed Central

    Jacobs, Chris G. C.; Rezende, Gustavo L.; Lamers, Gerda E. M.; van der Zee, Maurijn

    2013-01-01

    Insects have been extraordinarily successful in occupying terrestrial habitats, in contrast to their mostly aquatic sister group, the crustaceans. This success is typically attributed to adult traits such as flight, whereas little attention has been paid to adaptation of the egg. An evolutionary novelty of insect eggs is the serosa, an extraembryonic membrane that enfolds the embryo and secretes a cuticle. To experimentally test the protective function of the serosa, we exploit an exceptional possibility to eliminate this membrane by zerknüllt1 RNAi in the beetle Tribolium castaneum. We analyse hatching rates of eggs under a range of humidities and find dramatically decreasing hatching rates with decreasing humidities for serosa-less eggs, but not for control eggs. Furthermore, we show serosal expression of Tc-chitin-synthase1 and demonstrate that its knock-down leads to absence of the serosal cuticle and a reduction in hatching rates at low humidities. These developmental genetic techniques in combination with ecological testing provide experimental evidence for a crucial role of the serosa in desiccation resistance. We propose that the origin of this extraembryonic membrane facilitated the spectacular radiation of insects on land, as did the origin of the amniote egg in the terrestrial invasion of vertebrates. PMID:23782888

  12. Homochiral Asymmetric Triacylglycerol Isomers in Egg Yolk.

    PubMed

    Nagai, Toshiharu; Ishikawa, Keiko; Yoshinaga, Kazuaki; Yoshida, Akihiko; Beppu, Fumiaki; Gotoh, Naohiro

    2017-12-01

    The composition of triacylglycerol (TAG) positional isomer (-PI) and enantiomer (-E) in immature chicken egg yolk, mature chicken yolk, and chicken meat was examined. POO (consisting of one palmitic acid (P) and two oleic acids (Os)), PPO (consisting of two Ps and one O), and PPL (consisting of two Ps and one linoleic acid (L)) were treated as representative TAG molecular species in all the analytical samples because P, O, and L were the major fatty acids comprising egg and chicken meat. sn-POO (binding P at sn-1 position) was predominant in egg yolks, while sn-OOP and sn-OPO were present in chicken meat. This difference was ascribed to the different roles of these isomers as nutrients, because TAG in egg yolk is important for new born organisms and TAG in chicken meat is used for fat accumulation. The compositions of the TAG isomers in PPO and PPL in egg yolk were similar, and O and L did not bind at the sn-1 position. In contrast, all the isomers of PPO and PPL were found in chicken meat. These results imply that the TAG structure could be modified so that the nutrient requirement is fulfilled in egg yolk and chicken meat.

  13. Egg hatch and survival and development of beet webworm (Lepidoptera: Crambidae) larvae at different combinations of temperature and relative humidity

    USDA-ARS?s Scientific Manuscript database

    To understand the role that temperature and humidity play in the population dynamics of the beet webworm, Loxostege sticticalis L. (Lepidoptera: Crambidae), egg hatchability, survival of 1st - 5th instars, survival of the complete larval stage, survival curves, and larval development rates were inve...

  14. The role of emerging technologies to ensure the microbial safety of fresh produce, milk and eggs

    USDA-ARS?s Scientific Manuscript database

    This article reviews emerging techniques that are applied in the produce and dairy industry to ensure product safety. Microbial safety of produce, dairy and egg continues to be a major concern. According to Economic Research Service, USDA the cost of foodborne illnesses in the U.S. tops $15.6 billio...

  15. Antiinflammatory and analgesic effects of Psidium guajava Linn. (Myrtaceae) leaf aqueous extract in rats and mice.

    PubMed

    Ojewole, J A O

    2006-09-01

    In many parts of Africa, the leaf, stem-bark, and roots of Psidium guajava Linn. (Family: Myrtaceae) are used traditionally for the management, control, and/or treatment of an array of human disorders. In an effort to scientifically appraise some of the ethnomedical properties of P. guajava leaf, and probe its efficacy and safety, the present study was undertaken to examine the antiinflammatory and analgesic properties of the plant's leaf aqueous extract in some experimental animal paradigms. The antiinflammatory property of the aqueous leaf extract was investigated in rats, using fresh egg albumin-induced pedal (paw) edema, while the analgesic effect of the plant extract was evaluated by the "hot-plate" and "acetic acid" test models of pain in mice. Diclofenac (100 mg/kg, i.p.) and morphine (10 mg/kg, i.p.) were used respectively as standard, reference antiinflammatory and analgesic agents for comparison. P. guajava leaf aqueous extract (PGE, 50-800 mg/kg, i.p.) produced dose-dependent and significant (p < 0.05-0.001) inhibition of fresh egg albumin-induced acute inflammation (edema) in rats. The plant extract (PGE, 50-800 mg/kg, i.p.) also produced dose-dependent and significant (p < 0.05-0.001) analgesic effects against thermally and chemically induced nociceptive pain in mice. The numerous tannins, polyphenolic compounds, flavonoids, ellagic acid, triterpenoids, guiajaverin, quercetin, and other chemical compounds present in the plant are speculated to account for the observed antiinflammatory and analgesic effects of the plant's leaf extract. In summary, the findings of this experimental animal study indicate that the leaf aqueous extract of P. guajava possesses analgesic and antiinflammatory properties, and thus lend pharmacological credence to the suggested ethnomedical, folkloric uses of the plant in the management and/or control of painful, arthritic and other inflammatory conditions in some rural communities of Africa. (c) 2006 Prous Science. All rights reserved.

  16. Laboratory Evaluations of the Fractions Efficacy of Annona senegalensis (Annonaceae) Leaf Extract on Immature Stage Development of Malarial and Filarial Mosquito Vectors.

    PubMed

    Lame, Younoussa; Nukenine, Elias Nchiwan; Pierre, Danga Yinyang Simon; Elijah, Ajaegbu Eze; Esimone, Charles Okechukwu

    2015-12-01

    Within the framework to control mosquitoes, ovicidal, larvicidal and pupicidal activity of Annona senegalensis leaf extract and its 4 fractions against Anopheles gambiae and Culex quinquefasciatus were evaluated in the laboratory conditions. Ovicidal test was performed by submitting at least 100 eggs of mosquitoes to 125, 250, 500, 1000 and 2000 ppm concentrations, while larvicidal and pupicidal effects were assessed by submitting 25 larvae or pupae to the concentrations of 2500, 1250, 625 and 312.5 ppm of plant extract or fractions of A. senegalensis. The eggs of An. gambiae were most affected by N-hexane (0.00% hatchability) and chloroform (03.67% hatchability) fractions compared to Cx. quinquefasciatus where at least 25 % hatchability were recorded at 2000 ppm. For larvicidal test, N-hexane (LC50= 298.8 ppm) and chloroform (LC50= 418.3 ppm) fractions were more effective than other fractions on An. gambiae larvae while, a moderate effectiveness was also observed with N-hexane (LC50= 2087.6 ppm), chloroform (LC50= 9010.1 ppm) fractions on Cx. quinquefasciatus larvae. The highest mortality percent of the pupae were also recorded with N-hexane and chloroform fractions on An. gambiae at 2500 ppm. As for Cx. quinquefasciatus only 50 % and 36 % mortality were recorded with N-hexane and chloroform fractions respectively. The extract of A. senegalensis was toxic on immature stage of mosquito species tested. By splitting methanolic crude extract, only N-hexane and chloroform fractions were revealed to possess a mosquitocidal effects and could be considered and utilized for future immature mosquito vectors control.

  17. Effects of Temperature on In Vitro Short-Term Storage of Sterlet Sturgeon (Acipenser Ruthenus) Ova.

    PubMed

    Linhart, O; Shelton, W L; Tučková, V; Rodina, M; Siddique, Mam

    2016-02-01

    Artificial propagation of sturgeons is becoming increasingly important for recovery efforts as well as for commercial production. Sterlet Acipenser ruthenus is a common Eurasian sturgeon with a small body size and one of the fastest reproductive cycles among the sturgeons. The practical question being addressed in this study was how long fertilization of ovulated eggs can be delayed without substantially reducing the hatching rate, and an ancillary question is under what' temperature conditions do eggs retain good quality. Broodstock were injected with homogenized carp pituitary extract (CPE); ovulated eggs from three females were allocated to various treatment groups for temperature storage (control, 7°C, 11°C, 15°C and 19°C) until fertilized. Storage times at the regulated temperatures prior to fertilization were for 2.5, 5.0, 7.5 and 10.0 h. After the selected storage times in ovarian fluid, eggs were fertilized and transferred to incubation cages and then they were counted. Three replicates were allocated to each storage period and temperature. Hatched larvae were counted at 7-day post-fertilization. We found that sterlet eggs do not need to be fertilized immediately after collection. Reasonably good quality was retained for several hours if temperature conditions are fairly cool and stable. Eggs retained good quality when stored at 7°C and 11°C for up to 10 h with 54.1 ± 2.9 to 69.9 ± 7.9% hatching success, but egg quality was significantly reduced after 5-h storage at 19°C (p < 0.01) and 7.5-h storage at 15°C (p < 0.05) compared to cooler temperatures. Uniform temperatures between 7°C and 11°C can be considered as appropriate for storage of eggs in ovarian fluid for up to 10 h. This information can have practical application to routine hatchery practice for acipenserids, as well as for certain research protocols. © 2015 Blackwell Verlag GmbH.

  18. Estrogenic activity and reproductive effects of the UV-filter oxybenzone (2-hydroxy-4-methoxyphenyl-methanone) in fish.

    PubMed

    Coronado, Michael; De Haro, Hector; Deng, Xin; Rempel, Mary Ann; Lavado, Ramon; Schlenk, Daniel

    2008-11-21

    Previous studies in extracts of sediments surrounding municipal outfalls off the coast of California, USA and effluents of New York City, NY, USA indicated the UV-filtering agent, oxybenzone (CAS# 131-57-7; benzophenone-3) as a potential estrogen. The effects of oxybenzone on estrogenic activity and reproduction were evaluated using a 14-day juvenile rainbow trout assay for plasma vitellogenin and a subsequent 21-day Japanese medaka reproduction assay. Significant induction of vitellogenin was observed in the rainbow trout at the 1000 microg/L nominal concentration (749 microg/L median measured value) of oxybenzone which was approximately 75 times greater than the concentrations observed in previous wastewater effluent. Vitellogenin induction was also observed in the 1000 microg/L nominal concentration (620 microg/L median measured) of oxybenzone in male Japanese medaka (Oryzias latipes) after 21 days of exposure. The number of eggs produced per female per day exposed to the same concentration (620 microg/L) were significantly lower after 7 days, but returned to control values after 21 days. Fertilized eggs were then monitored for 20 days to assess hatching success. The overall percentage of fertilized eggs collected during the 21-day exposure that hatched was significantly lower in the 620 microg/L oxybenzone concentration. There was also a temporal effect at this concentration as egg viability (percentage of fertilized eggs that hatched) was diminished 13-15 days after eggs were collected. All three oxybenzone concentrations (16, 132, and 620 microg/L) and the 50 ng/L estradiol positive control showed reduced hatching of eggs at day 15, and the 132 and 620 microg/L oxybenzone concentrations diminished the percentage of eggs that hatched on days 13-15. These data indicate that the UV-filter oxybenzone alters endocrine or reproduction endpoints in two fish species, but at concentrations significantly higher than those measured in the environment.

  19. Discrimination of gastrointestinal nematode eggs from crude fecal egg preparations by inhibitor-resistant conventional and real-time PCR.

    PubMed

    Demeler, Janina; Ramünke, Sabrina; Wolken, Sonja; Ianiello, Davide; Rinaldi, Laura; Gahutu, Jean Bosco; Cringoli, Giuseppe; von Samson-Himmelstjerna, Georg; Krücken, Jürgen

    2013-01-01

    Diagnosis of gastrointestinal nematodes relies predominantly on coproscopic methods such as flotation, Kato-Katz, McMaster or FLOTAC. Although FLOTAC allows accurate quantification, many nematode eggs can only be differentiated to genus or family level. Several molecular diagnostic tools discriminating closely related species suffer from high costs for DNA isolation from feces and limited sensitivity since most kits use only small amounts of feces (<1 g). A direct PCR from crude egg preparations was designed for full compatibility with FLOTAC to accurately quantify eggs per gram feces (epg) and determine species composition. Eggs were recovered from the flotation solution and concentrated by sieving. Lysis was achieved by repeated boiling and freezing cycles - only Trichuris eggs required additional mechanic disruption. Egg lysates were directly used as template for PCR with Phusion DNA polymerase which is particularly resistant to PCR inhibitors. Qualitative results were obtained with feces of goats, cattle, horses, swine, cats, dogs and mice. The finally established protocol was also compatible with quantitative real-time PCR in the presence of EvaGreen and no PCR inhibition was detectable when extracts were diluted at least fourfold. Sensitivity was comparable to DNA isolation protocols and spiked samples with five epg were reliably detected. For Toxocara cati a detection limit below one epg was demonstrated. It was possible to distinguish T. cati and Toxocara canis using high resolution melt (HRM) analysis, a rapid tool for species identification. In human samples, restriction fragment length polymorphism (RFLP) and HRM analysis were used to discriminate Necator americanus and Ancylostoma duodenale. The method is able to significantly improve molecular diagnosis of gastrointestinal nematodes by increasing speed and sensitivity while decreasing overall costs. For identification of species or resistance alleles, analysis of PCR products with many different post PCR methods can be used such as RFLP, reverse-line-blot, Sanger sequencing and HRM.

  20. Discrimination of Gastrointestinal Nematode Eggs from Crude Fecal Egg Preparations by Inhibitor-Resistant Conventional and Real-Time PCR

    PubMed Central

    Demeler, Janina; Ramünke, Sabrina; Wolken, Sonja; Ianiello, Davide; Rinaldi, Laura; Gahutu, Jean Bosco; Cringoli, Giuseppe; von Samson-Himmelstjerna, Georg; Krücken, Jürgen

    2013-01-01

    Diagnosis of gastrointestinal nematodes relies predominantly on coproscopic methods such as flotation, Kato-Katz, McMaster or FLOTAC. Although FLOTAC allows accurate quantification, many nematode eggs can only be differentiated to genus or family level. Several molecular diagnostic tools discriminating closely related species suffer from high costs for DNA isolation from feces and limited sensitivity since most kits use only small amounts of feces (<1 g). A direct PCR from crude egg preparations was designed for full compatibility with FLOTAC to accurately quantify eggs per gram feces (epg) and determine species composition. Eggs were recovered from the flotation solution and concentrated by sieving. Lysis was achieved by repeated boiling and freezing cycles – only Trichuris eggs required additional mechanic disruption. Egg lysates were directly used as template for PCR with Phusion DNA polymerase which is particularly resistant to PCR inhibitors. Qualitative results were obtained with feces of goats, cattle, horses, swine, cats, dogs and mice. The finally established protocol was also compatible with quantitative real-time PCR in the presence of EvaGreen and no PCR inhibition was detectable when extracts were diluted at least fourfold. Sensitivity was comparable to DNA isolation protocols and spiked samples with five epg were reliably detected. For Toxocara cati a detection limit below one epg was demonstrated. It was possible to distinguish T. cati and Toxocara canis using high resolution melt (HRM) analysis, a rapid tool for species identification. In human samples, restriction fragment length polymorphism (RFLP) and HRM analysis were used to discriminate Necator americanus and Ancylostoma duodenale. The method is able to significantly improve molecular diagnosis of gastrointestinal nematodes by increasing speed and sensitivity while decreasing overall costs. For identification of species or resistance alleles, analysis of PCR products with many different post PCR methods can be used such as RFLP, reverse-line-blot, Sanger sequencing and HRM. PMID:23620739

  1. Evaluation of antiviral activity of Ocimum sanctum and Acacia arabica leaves extracts against H9N2 virus using embryonated chicken egg model.

    PubMed

    Ghoke, S S; Sood, R; Kumar, N; Pateriya, A K; Bhatia, S; Mishra, A; Dixit, R; Singh, V K; Desai, D N; Kulkarni, D D; Dimri, U; Singh, V P

    2018-06-05

    In the view of endemic avian influenza H9N2 infection in poultry, its zoonotic potential and emergence of antiviral resistance, two herbal plants, Ocimum sanctum and Acacia arabica, which are easily available throughout various geographical locations in India were taken up to study their antiviral activity against H9N2 virus. We evaluated antiviral efficacy of three different extracts each from leaves of O. sanctum (crude extract, terpenoid and polyphenol) and A. arabica (crude extract, flavonoid and polyphenol) against H9N2 virus using in ovo model. The antiviral efficacy of different leaves extracts was systematically studied in three experimental protocols viz. virucidal (dose-dependent), therapeutic (time-dependent) and prophylactic (dose-dependent) activity employing in ovo model. The maximum non-toxic concentration of each herbal extracts of O. sanctum and A. arabica in the specific pathogen free embryonated chicken eggs was estimated and their antiviral efficacy was determined in terms of reduction in viral titres, measured by Haemagglutination (HA) and real time quantitative reverse transcription polymerase chain reaction (RT-qPCR) assays. All the extracts of O. sanctum (crude extract, terpenoid and polyphenol) and A. arabica (crude extract, flavonoid and polyphenol) showed significant virucidal activity, however, crude extract ocimum and terpenoid ocimum showed highly significant to significant (p < 0.001-0.01) decrease in virus genome copy numbers with lowest dose tested. Similarly, therapeutic effect was observed in all three extracts of O. sanctum in comparison to the virus control, nevertheless, crude extract ocimum and terpenoid ocimum maintained this effect for longer period of time (up to 72 h post-incubation). None of the leaves extracts of A. arabica had therapeutic effect at 24 and 48 h post-incubation, however, only the crude extract acacia and polyphenol acacia showed delayed therapeutic effect (72 h post-inoculation). Prophylactic potential was observed in polyphenol acacia with highly significant antiviral activity compared to virus control (p < 0.001). The crude extract and terpenoid isolated from the leaves of O. sanctum and polyphenol from A. arabica has shown promising antiviral properties against H9N2 virus. Future investigations are necessary to formulate combinations of these compounds for the broader antiviral activity against H9N2 viruses and evaluate them in chickens.

  2. A chitin-like component in Aedes aegypti eggshells, eggs and ovaries.

    PubMed

    Moreira, Mônica F; Dos Santos, Amanda S; Marotta, Humberto R; Mansur, Juliana F; Ramos, Isabela B; Machado, Ednildo A; Souza, Gustavo H M F; Eberlin, Marcos N; Kaiser, Carlos R; Kramer, Karl J; Muthukrishnan, Subbaratnam; Vasconcellos, Ana Maria H

    2007-12-01

    An insoluble white substance was prepared from extracts of eggshells of Aedes aegypti, the yellow fever mosquito and dengue vector. Its infrared and proton NMR spectra were similar to that of standard commercial chitin. This putative chitin-like material, also obtained from ovaries, newly laid and dark eggs, was hydrolyzed in acid and a major product was identified by HPLC to be glucosamine. The eggshell acid hydrolysate was also analyzed by ESI-MS and an ion identical to a glucosamine monoprotonated species was detected. The presence of chitin was also analyzed during different developmental stages of the ovary using a fluorescent microscopy technique and probes specific for chitin. The results showed that a chitin-like material accumulates in oocytes during oogenesis. Streptomyces griseus chitinase pre-treatment of oocytes greatly reduced the chitin-derived fluorescence. Chitinase activity was detected in newborn larvae and eggs prior to hatching. Feeding experiments indicated that the chitin synthesis inhibitor lufenuron inhibited chitin synthesis, either when mosquitoes were allowed to feed directly on lufenuron-treated chickens or when an artificial feeding system was used. Lufenuron inhibited egg hatch, larval development and reduced mosquito viability. These data demonstrate for the first time that (1) a chitin-like material is present in A. aegypti eggs, ovaries and eggshells; (2) a chitin synthesis inhibitor can be used to inhibit mosquito oogenesis; and (3) chitin synthesis inhibitors have potential for controlling mosquito populations.

  3. Bone sialoprotein keratan sulfate proteoglycan (BSP-KSPG) and FGF-23 are important physiological components of medullary bone.

    PubMed

    Hadley, Jill A; Horvat-Gordon, Maria; Kim, Woo-Kyun; Praul, Craig A; Burns, Dennis; Leach, Roland M

    2016-04-01

    Medullary bone is a specialized bone found in the marrow cavity of laying birds. It provides a significant contribution to the calcium supply for egg shell formation. Medullary bone is distinguished from cortical bone by the presence of large amounts of a keratan sulfate proteoglycan (KSPG). The aims of the present experiment are to confirm the identity of the core protein of KSPG, identify a marker of medullary bone metabolism, and determine whether changes in keratan sulfate (KS) concentration in blood are associated with the egg-laying cycle. Using two different isolation techniques- one specific for bone and another for blood- we have identified bone sialoprotein (BSP) to be the core protein of this KSPG. We also determined that the amount of keratan sulfate (KS) in laying hen blood fluctuates in synchrony with the egg-laying cycle, and thus can serve as a specific marker for medullary bone metabolism. During the course of this investigation, we also found FGF-23 (phosphatonin) to be expressed in medullary bone, in synchrony with the egg-laying cycle. Western blotting was used to demonstrate the presence of this peptide in both laying hen blood and medullary bone extracts. The importance of FGF-23 (phosphatonin) and parathyroid hormone in normalizing the dramatic changes in plasma calcium and phosphorus during the 24h egg-laying cycle is discussed. Copyright © 2016 Elsevier Inc. All rights reserved.

  4. Anthelmintic activity of Chenopodium album (L) and Caesalpinia crista (L) against trichostrongylid nematodes of sheep.

    PubMed

    Jabbar, Abdul; Zaman, Muhammad Arfan; Iqbal, Zafar; Yaseen, Muhammad; Shamim, Asim

    2007-10-08

    The present study was carried out to determine the anthelmintic activity of Caesalpinia crista (L.) (Fabaceae) seed kernel and Chenopodium album (L.) (Chenopodiaceae) whole plant in order to justify their traditional use in veterinary medicine. In vitro anthelmintic activity of crude aqueous methanolic extract (AME) of both the plants was determined using mature Haemonchus contortus and their eggs in adult motility assay and egg hatch test, respectively. In vivo anthelmintic activity was evaluated in sheep naturally infected with mixed species of gastrointestinal nematodes by administering crude powder (CP) and AME in increasing doses (1.0-3.0 g/kg). Both plants exhibited dose- and time-dependent anthelmintic effects by causing mortality of worms and inhibition of egg hatching. Caesalpinia crista (LC50=0.134 mg/mL) was found to be more potent than Chenopodium album (LC50=0.449 mg/mL) in egg hatch test. In vivo, maximum reduction in eggs per gram (EPG) of faeces was recorded as 93.9 and 82.2% with Caesalpinia crista and Chenopodium album AME at 3.0 g/kg on day 13 and 5 post-treatment, respectively. Levamisole (7.5 mg/kg), a standard anthelmintic agent, showed 95.1-95.6% reduction in EPG. These data show that both Caesalpinia crista and Chenopodium album possess anthelmintic activity in vitro and in vivo, thus, justifying their use in the traditional medicine system of Pakistan.

  5. Enrichment of conjugated linoleic acid (CLA) in hen eggs and broiler chickens meat by lactic acid bacteria.

    PubMed

    Herzallah, Saqer

    2013-01-01

    1. The aim of this work was to compare conjugated linoleic acid (CLA) concentrations in chickens supplemented with 4 American Tissue Culture Collection (ATCC) bacterial strains, Lactobacillus plantarum, Lactobacillus lactis, Lactobacillus casei and Lactobacillus fermentum, and 4 isolates of Lactobacillus reuteri from camel, cattle, sheep and goat rumen extracts. 2. Micro-organisms were grown anaerobically in MRS broth, and 10(6) CFU/ml of bacteria were administered orally to mixed-sex, 1-d-old broiler chickens weekly for 4 weeks and to 23-week-old layer hens weekly for 6 weeks. 3. The 4 strains were evaluated for their effects on synthesis of CLA in hen eggs and broiler meat cuts. 4. Administration of pure Lactobacillus and isolated L. reuteri strains from camel, cattle, goat and sheep led to significantly increased CLA concentrations of 0.2-1.2 mg/g of fat in eggs and 0.3-1.88 mg/g of fat in broiler chicken flesh homogenates of leg, thigh and breast. 5. These data demonstrate that lactic acid bacteria of animal origin (L. reuteri) significantly enhanced CLA synthesis in both eggs and broiler meat cuts.

  6. A novel approach for food intake detection using electroglottography

    PubMed Central

    Farooq, Muhammad; Fontana, Juan M; Sazonov, Edward

    2014-01-01

    Many methods for monitoring diet and food intake rely on subjects self-reporting their daily intake. These methods are subjective, potentially inaccurate and need to be replaced by more accurate and objective methods. This paper presents a novel approach that uses an Electroglottograph (EGG) device for an objective and automatic detection of food intake. Thirty subjects participated in a 4-visit experiment involving the consumption of meals with self-selected content. Variations in the electrical impedance across the larynx caused by the passage of food during swallowing were captured by the EGG device. To compare performance of the proposed method with a well-established acoustical method, a throat microphone was used for monitoring swallowing sounds. Both signals were segmented into non-overlapping epochs of 30 s and processed to extract wavelet features. Subject-independent classifiers were trained using Artificial Neural Networks, to identify periods of food intake from the wavelet features. Results from leave-one-out cross-validation showed an average per-epoch classification accuracy of 90.1% for the EGG-based method and 83.1% for the acoustic-based method, demonstrating the feasibility of using an EGG for food intake detection. PMID:24671094

  7. Role of tetraspanin CD9 molecule in fertilization of mammals.

    PubMed

    Jankovičová, J; Simon, M; Antalíková, J; Cupperová, P; Michalková, K

    2015-01-01

    Fertilization process is a very clever and unique process comprising some essential steps resulting in formation of zygote. Tetraspanin CD9 is considered to be a serious candidate molecule participating in these events. The importance of CD9 has been discussed in relation to acrosome reaction, sperm-binding, sperm-penetration, sperm-egg fusion and eventually, egg activation. The abundant expression of CD9 oocyte plasma membrane and the presence of CD9-containing vesicles in the perivitelline space of intact oocytes have been confirmed. Despite the fact that majority of authors analyzed CD9 expressed on oocytes, several studies considered the function of sperm CD9, too. To understand CD9 involvement, various conditions of in vitro fertilization (IVF) assays using polyclonal as well as monoclonal antibodies or knockout mice were carried out. However, ambiguous data have been obtained about the importance of CD9 in sperm-egg binding or fusion. Although the current findings did not prove any hypothesis, the indispensable role of CD9 in fertilization process was not excluded and the precise role of CD9 remains unexplained.

  8. A flotation/sieving method to detect Echinococcus multilocularis and Toxocara spp. eggs in soil by real-time PCR

    PubMed Central

    Umhang, Gérald; Bastien, Matthieu; Renault, Camille; Faisse, Marine; Caillot, Christophe; Boucher, Jean-Marc; Hormaz, Vanessa; Poulle, Marie-Lazarine; Boué, Franck

    2017-01-01

    Soil can be a source of human infection by many zoonotic helminth species including Echinococcus multilocularis and Toxocara spp. The prevention of alveolar echinococcosis could be greatly improved through the identification of at-risk areas. Yet very few data are available about the detection of E. multilocularis in soil, while more studies have been reported for Toxocara spp. Identification of soil contamination by E. multilocularis eggs requires the use of specific methods. This study describes the development of a method for the detection of E. multilocularis in soil samples with the concentration of eggs using a flotation/sieving method and detection by duplex real-time polymerase chain reaction (PCR). Toxocara spp. egg detection was also undertaken due to the widespread presence of this parasite in soil, despite it being considered less pathogenic. Method sensitivity of 100% was reached for the detection of 10 E. multilocularis eggs spiked in 10 g of soil. Concerning Toxocara spp., method sensitivity was lower but assumed to be due to the reduced effectiveness of the DNA extraction protocol. The parasitological status for E. multilocularis and Toxocara spp. of 63 carnivore fecal samples collected in highly endemic rural areas of France and of soil samples collected under and near these fecal samples was compared. The contamination of soil samples collected under positive fecal samples for E. multilocularis (n = 3) or Toxocara spp. (n = 19) confirmed the transfer of eggs from the definitive host to the environment. PMID:28737135

  9. Dioxins and PCBs in ostrich meat and eggs: levels and implications.

    PubMed

    Piskorska-Pliszczynska, Jadwiga; Strucinski, Pawel; Mikolajczyk, Szczepan; Pajurek, Marek; Maszewski, Sebastian; Pietron, Wojciech

    2017-12-01

    Although consumption of eggs is an essential part of our diet, limited information is available for table eggs other than those laid by hens. The aim of our study was to determine concentrations of polychlorinated dibenzofurans (PCDD/Fs), dioxin-like (DL-) and non-dioxin-like (NDL) polychlorinated biphenyls (PCBs) in ostrich eggs and meat available on the Polish market, in order to obtain baseline information on the current status of these pollutants in comparison to poultry products. Obtained data were compared with the binding EU limits set for chicken eggs and meat. The levels of individual PCDD/Fs and PCBs congeners varied considerably. The percentage share of total WHO toxic equivalency factor (WHO-TEQ) content indicates the dominant role of PCDD/Fs. High concentrations of PCDD/F and DL-PCBs, in the range of 0.85-74.48 pg WHO-TEQ g -1 fat, were found in ostrich eggs; this exceeds the maximum level permitted for chicken eggs by a factor of up to 15. Eight of the 11 egg samples exceeded the action level for hen eggs. Although the ostrich meat concentrations of PCDD/Fs do not exceed the limit established for poultry muscle (1.75 pg g -1 fat), average contents of PCDD/Fs exceeded almost four times the levels in chicken and turkey muscle. Human exposure was evaluated and the resulting risk was characterised. Taking into account the low average consumption of ostrich eggs, the resulting exposure to dioxins for the general population can be considered as negligibly low. However, the individuals who frequently consume such eggs may be at risk of elevated exposure. Although ostrich products are not consumed frequently, such data are nevertheless useful for food safety purposes.

  10. Differential gene expression during early embryonic development in diapause and non-diapause eggs of multivoltine silkworm Bombyx mori.

    PubMed

    Ponnuvel, Kangayam M; Murthy, Geetha N; Awasthi, Arvind K; Rao, Guruprasad; Vijayaprakash, Nanjappa B

    2010-11-01

    Quantification of the differential expression of metabolic enzyme and heat-shock protein genes (Hsp) during early embryogenesis in diapause and non-diapause eggs of the silkworm B. mori was carried out by semi-quantitative RT-PCR. Data analysis revealed that, the phosphofructokinase (PFK) expression started at a higher level in the early stage (6 h after oviposition) in non-diapause eggs, while in diapause induced eggs, it started at a lower level. However, the PFK gene expression in diapause eggs was comparatively higher than in non-diapause eggs. PFK facilitates use of carbohydrate reserves. The lower level of PFK gene expression in the early stage of diapause induced eggs but comparatively higher level of expression than in non-diapause eggs is due to enzyme inactivation via protein phosphorylation during early embryogenesis followed by de-phosphorylation in later stage. The sorbitol dehydrogenase-2 (SDH-2) gene was down regulated in diapause induced eggs up to 24 h and its expression levels in diapause induced eggs coincided with that of PFK gene at 48h in non-diapause eggs. During carbohydrate metabolism, there is an initial temporary accumulation of sorbitol which acts as protectant. The down regulation of SDH-2 gene during the first 24 hours in diapause induced eggs was due to the requirement of sorbitol as protectant. However, since the diapause process culminates by 48 h, the SDH-2 gene expression increased and coincided with that of PFK gene expression. The trehalase (Tre) gene expression was at a lower level in diapause induced eggs compared to non-diapausing eggs. The induction of Tre activity is to regulate uptake and use of sugar by the tissues. The non-diapause eggs revealed maximum expression of GPase gene with major fluctuations as well as an overall higher expression compared to diapause induced eggs. The diapause process requires less energy source which reflects lower activity of the gene. Heat shock protein (Hsp) genes (Hsp20.4, 40, 70, and 90) revealed differential levels of expression in both the eggs at all stages of embryonic development. The present study thus provides an overview of the differential expression levels of metabolic enzyme and Hsp genes in non-diapause and diapause induced eggs of multivoltine silkworm B. mori within 48 h after oviposition, confirming the major role of in early embryogenesis.

  11. A RAPID AND SIMPLE PHOSPHOLIPASE A ASSAY,

    DTIC Science & Technology

    A simple and rapid method for the assay of phospholipase A was developed. As a substrate fresh egg yolk is used which is hydrolyzed by snake venom...phospholipase A at a 10-20 x faster rate than pure lecithin . The released fatty acids, after extraction with appropriate solvents are titrated

  12. Unfertilized frog eggs die by apoptosis following meiotic exit

    PubMed Central

    2011-01-01

    Background A characteristic feature of frog reproduction is external fertilization accomplished outside the female's body. Mature fertilization-competent frog eggs are arrested at the meiotic metaphase II with high activity of the key meiotic regulators, maturation promoting factor (MPF) and cytostatic factor (CSF), awaiting fertilization. If the eggs are not fertilized within several hours of ovulation, they deteriorate and ultimately die by as yet unknown mechanism. Results Here, we report that the vast majority of naturally laid unfertilized eggs of the African clawed frog Xenopus laevis spontaneously exit metaphase arrest under various environmental conditions and degrade by a well-defined apoptotic process within 48 hours after ovulation. The main features of this process include cytochrome c release, caspase activation, ATP depletion, increase of ADP/ATP ratio, apoptotic nuclear morphology, progressive intracellular acidification, and egg swelling. Meiotic exit seems to be a prerequisite for execution of the apoptotic program, since (i) it precedes apoptosis, (ii) apoptotic events cannot be observed in the eggs maintaining high activity of MPF and CSF, and (iii) apoptosis in unfertilized frog eggs is accelerated upon early meiotic exit. The apoptotic features cannot be observed in the immature prophase-arrested oocytes, however, the maturation-inducing hormone progesterone renders oocytes susceptible to apoptosis. Conclusions The study reveals that naturally laid intact frog eggs die by apoptosis if they are not fertilized. A maternal apoptotic program is evoked in frog oocytes upon maturation and executed after meiotic exit in unfertilized eggs. The meiotic exit is required for execution of the apoptotic program in eggs. The emerging anti-apoptotic role of meiotic metaphase arrest needs further investigation. PMID:22195698

  13. Paradigm shift in the management of milk and egg allergy: baked milk and egg diet.

    PubMed

    Konstantinou, George N; Kim, Jennifer S

    2012-02-01

    Heat treatment of several foods, including all types of cooking, has been mainly used to minimize the number of viable microbes, reduce pathogenicity, and destroy the undesirable enzymes, maintaining food quality. In addition, food processing improves sensory, nutritional, and physical properties of the foods, due to food protein denaturation. Heat-induced alterations of food proteins can attenuate allergenicity. In this article, the authors review the important role of thermal processing on milk and egg proteins, which comprise the commonest food allergies in infancy and early childhood. Copyright © 2012 Elsevier Inc. All rights reserved.

  14. Role of specific IgE and skin-prick testing in predicting food challenge results to baked egg

    PubMed Central

    Cortot, Catherine F.; Sheehan, William J.; Permaul, Perdita; Friedlander, James L.; Baxi, Sachin N.; Gaffin, Jonathan M.; Dioun, Anahita F.; Hoffman, Elaine B.; Schneider, Lynda C.

    2012-01-01

    Previous studies suggest that children with egg allergy may be able to tolerate baked egg. Reliable predictors of a successful baked egg challenge are not well established. We examined egg white–specific IgE levels, skin-prick test (SPT) results, and age as predictors of baked egg oral food challenge (OFC) outcomes. We conducted a retrospective chart review of children, aged 2–18 years, receiving an egg white–specific IgE level, SPT, and OFC to baked egg from 2008 to 2010. Fifty-two oral baked egg challenges were conducted. Of the 52 challenges, 83% (n = 43) passed and 17% (n = 9) failed, including 2 having anaphylaxis. Median SPT wheal size was 12 mm (range, 0–35 mm) for passed challenges and 17 mm (range, 10–30 mm) for failed challenges (p = 0.091). The negative predictive value for passing the OFC was 100% (9 of 9) if SPT wheal size was <10 mm. Median egg white–specific IgE was 2.02 kU/L (range, <0.35–13.00 kU/L) for passed challenges and 1.52 kU/L (range, 0.51–6.10 kU/L) for failed challenges (p = 0.660). Receiver operating characteristic (ROC) curve analysis for SPT revealed an area under the curve (AUC) of 0.64. ROC curve analysis for egg white–specific IgE revealed an AUC of 0.63. There was no significant difference in age between patients who failed and those who passed (median = 8.8 years versus 7.0 years; p = 0.721). Based on our sample, SPT, egg white–specific IgE and age are not good predictors of passing a baked egg challenge. However, there was a trend for more predictability with SPT wheal size. PMID:22584194

  15. The Genetic Basis of Natural Variation in Drosophila (Diptera: Drosophilidae) Virgin Egg Retention

    PubMed Central

    Akhund-Zade, Jamilla; Bergland, Alan O.; Crowe, Sarah O.; Unckless, Robert L.

    2017-01-01

    Drosophila melanogaster is able to thrive in harsh northern climates through adaptations in life-history traits and physiological mechanisms that allow for survival through the winter. We examined the genetic basis of natural variation in one such trait, female virgin egg retention, which was previously shown to vary clinally and seasonally. To further our understanding of the genetic basis and evolution of virgin egg retention, we performed a genome-wide association study (GWAS) using the previously sequenced Drosophila Genetic Reference Panel (DGRP) mapping population. We found 29 single nucleotide polymorphisms (SNPs) associated with virgin egg retention and assayed 6 available mutant lines, each harboring a mutation in a candidate gene, for effects on egg retention time. We found that four out of the six mutant lines had defects in egg retention time as compared with the respective controls: mun, T48, Mes-4, and Klp67A. Surprisingly, none of these genes has a recognized role in ovulation control, but three of the four genes have known effects on fertility or have high expression in the ovaries. We also found that the SNP set associated with egg retention time was enriched for clinal SNPs. The majority of clinal SNPs had alleles associated with longer egg retention present at higher frequencies in higher latitudes. Our results support previous studies that show higher frequency of long retention times at higher latitude, providing evidence for the adaptive value of virgin egg-retention. PMID:28042107

  16. Vegetation-derived cues for the selection of oviposition substrates by Anopheles albimanus under laboratory conditions.

    PubMed

    Torres-Estrada, José Luis; Meza-Alvarez, R Amanda; Cibrián-Tovar, Juan; Rodríguez-López, Mario H; Arredondo-Jiménez, Juan I; Cruz-López, Leopoldo; Rojas-Leon, Julio C

    2005-12-01

    Oviposition response of gravid Anopheles albimanus Wiedemman (Diptera: Culicidae) females to water containing Brachiaria mutica, Cynodon dactylon, Jouvea straminea, Fimbristylis spadicea, and Ceratophyllum demersum was investigated. Gravid An. albimanus females deposited similar egg numbers in cups containing natural plants in water from natural breeding sites and in cups containing natural plants in distilled water. Gravid mosquitoes deposited significantly more eggs in cups containing natural plants in water from natural breeding sites than in cups containing artificial plants in water from the corresponding natural breeding sites. These results were confirmed in experiments conducted in a wind tunnel, indicating that female response is mediated by chemical cues from plants. Bioassays with organic extracts of all 5 plant species indicated that these extracts at 100%, 10%, and 1% concentrations had an oviposition repellent effect, while attractiveness was observed at 0.1%, 0.01%, and 0.001%. Gas chromatography and mass spectrometry analysis of the organic extracts found in all 5 plants showed a mixture of terpenoid and alcohol compounds, among them: guaiacol, phenol, isoeugenol, longifolene, caryophyllene, phenyl ethyl alcohol, and p-cresol. These results suggest that middle-range volatiles from plants may function as chemical cues for the female's oviposition response in this mosquito species.

  17. Egg ovotransferrin-derived ACE inhibitory peptide IRW increases ACE2 but decreases proinflammatory genes expression in mesenteric artery of spontaneously hypertensive rats.

    PubMed

    Majumder, Kaustav; Liang, Guanxiang; Chen, Yanhong; Guan, LeLuo; Davidge, Sandra T; Wu, Jianping

    2015-09-01

    Egg ovotransferrin-derived angiotensin converting enzyme (ACE) inhibitory peptide IRW was previously shown to reduce blood pressure in spontaneously hypertensive rats through reduced vascular inflammation and increased nitric oxide-mediated vasorelaxation. The main objective of the present study was to investigate the molecular mechanism of this peptide through transcriptome analysis by RNAseq technique. Total RNA was extracted from kidney and mesenteric arteries; the RNAseq libraries (from untreated and IRW-treated groups) were constructed and subjected to sequence using HiSeq 2000 system (Illumina) system. A total of 12 764 and 13 352 genes were detected in kidney and mesenteric arteries, respectively. The differentially expressed (DE) genes between untreated and IRW-treated groups were identified and the functional analysis through ingenuity pathway analysis revealed a greater role of DE genes identified from mesenteric arteries than that of kidney in modulating various cardiovascular functions. Subsequent qPCR analysis further confirmed that IRW significantly increased the expression of ACE-2, ABCB-1, IRF-8, and CDH-1 while significantly decreased the expression ICAM-1 and VCAM-1 in mesenteric arteries. Our research showed for the first time that ACE inhibitory peptide IRW could contribute to its antihypertensive activity through increased ACE2 and decreased proinflammatory genes expression. © 2015 The Authors. Molecular Nutrition & Food Research published by Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

  18. Subcellular components of the amphibian egg - Insights provided by gravitational studies

    NASA Technical Reports Server (NTRS)

    Neff, A. W.; Ritzenthaler, J. D.; Rosenbaum, J. F.

    1989-01-01

    The variability in the response of Xenopus laevis eggs to a given force environment is studied. The roles of cytoplasmic organelle, the yolk platelets, and cytoskeletal components in varying in cytoplasmic mobility are examined. The data reveal that the packing of yolk platelets is not a major factor in causing cytoplasmic mobility differences and microtubules may affect cytoplasmic mobility.

  19. DoD Role for Securing United States Cyberspace

    DTIC Science & Technology

    2008-03-01

    of Agriculture is responsible for agriculture and food ( meat , poultry , and egg products). 2 The Department of Health and Human Services is...responsible for food other than meat , poultry , and egg products. 3 Nothing in this plan impairs or otherwise affects the authority of the Secretary of Defense...22  3.4.  Literature Review Content Analysis ..............................................................25  vi vii 3.5

  20. Duplex quantitative real-time PCR assay for the detection and discrimination of the eggs of Toxocara canis and Toxocara cati (Nematoda, Ascaridoidea) in soil and fecal samples

    PubMed Central

    2012-01-01

    Background Toxocarosis is a zoonotic disease caused by Toxocara canis (T. canis) and/or Toxocara cati (T. cati), two worldwide distributed roundworms which are parasites of canids and felids, respectively. Infections of humans occur through ingestion of embryonated eggs of T. canis or T. cati, when playing with soils contaminated with dogs or cats feces. Accordingly, the assessment of potential contamination of these areas with these roundworms eggs is paramount. Methods A duplex quantitative real-time PCR (2qPCR) targeting the ribosomal RNA gene internal transcribed spacer (ITS2) has been developed and used for rapid and specific identification of T. canis and T. cati eggs in fecal and soil samples. The assay was set up on DNA samples extracted from 53 adult worms including T. canis, T. cati, T. leonina, Ascaris suum (A. suum) and Parascaris equorum (P. equorum). The assay was used to assess the presence of T. cati eggs in several samples, including 12 clean soil samples spiked with eggs of either T. cati or A. suum, 10 actual soil samples randomly collected from playgrounds in Brussels, and fecal samples from cats, dogs, and other animals. 2qPCR results on dogs and cats fecal samples were compared with results from microscopic examination. Results 2qPCR assay allowed specific detection of T. canis and T. cati, whether adult worms, eggs spiked in soil or fecal samples. The 2qPCR limit of detection (LOD) in spiked soil samples was 2 eggs per g of soil for a turnaround time of 3 hours. A perfect concordance was observed between 2qPCR assay and microscopic examination on dogs and cats feces. Conclusion The newly developed 2qPCR assay can be useful for high throughput prospective or retrospective detection of T.canis and/or T. cati eggs in fecal samples as well as in soil samples from playgrounds, parks and sandpits. PMID:23216873

  1. Magnetic dispersive solid-phase extraction based on graphene oxide/Fe3 O4 @polythionine nanocomposite followed by atomic absorption spectrometry for zinc monitoring in water, flour, celery and egg.

    PubMed

    Babaei, Azar; Zeeb, Mohsen; Es-Haghi, Ali

    2018-07-01

    Magnetic graphene oxide nanocomposite has been proposed as a promising and sustainable sorbent for the extraction and separation of target analytes from food matrices. Sample preparation based on nanocomposite presents several advantages, such as desired efficiency, reasonable selectivity and high surface-area-to-volume ratio. A new graphene oxide/Fe 3 O 4 @polythionine (GO/Fe 3 O 4 @PTh) nanocomposite sorbent was introduced for magnetic dispersive solid-phase extraction and flame atomic absorption spectrometric detection of zinc(II) in water, flour, celery and egg. To fabricate the sorbent, an oxidative polymerization of thionine on the surface of magnetic GO was applied, while polythionine was simply employed as a surface modifier to improve extraction yield. The properties of the sorbent were characterized by transmission electron microscopy, scanning electron microscopy, X-ray diffraction, energy-dispersive X-ray analysis, vibrating sample magnetometry and Fourier transform-infrared spectroscopy. The calibration curve showed linearity in the range of 0.5-30 ng mL -1 . Limits of detection (S/N = 3) and quantification (S/N = 10) were 0.08 and 0.5 ng mL -1 , respectively. The method was applied for trace-level determination of Zn(II) in water and food samples, and its validation was investigated by recovery experiments and analyzing certified reference material. © 2018 Society of Chemical Industry. © 2018 Society of Chemical Industry.

  2. Diagnostic stages of the parasites of the Florida manatee, Trichechus manatus latirostris.

    PubMed

    Bando, Monica; Larkin, Iskande V; Wright, Scott D; Greiner, Ellis C

    2014-02-01

    Limited information is available on diagnostic stages of parasites in Florida manatees (Trichechus manatus latirostris). We examined 67 fecal samples from captive and wild manatees to define the diagnostic stages of the parasite fauna known to occur in Florida manatees. Parasite eggs were freshly extracted ex utero from identified mature helminths and subsequently characterized, illustrated, and matched to those isolated from fecal samples. In addition, coccidian oocysts in the fecal samples were identified. These diagnostic stages included eggs from 5 species of trematodes (Chiorchis fabaceus, Chiorchis groschafti, Pulmonicola cochleotrema, Moniligerum blairi, and Nudacotyle undicola), 1 nematode (Heterocheilus tunicatus), and oocysts of 2 coccidians (Eimeria manatus and Eimeria nodulosa).

  3. Efficacy of indigenous plant extracts on the malaria vector Anopheles subpictus Grassi (Diptera: Culicidae)

    PubMed Central

    Elango, G.; Zahir, A. Abduz; Bagavan, A.; Kamaraj, C.; Rajakumar, G.; Santhoshkumar, T.; Marimuthu, S.; Rahuman, A. Abdul

    2011-01-01

    Background & objectives: Mosquito control is facing a threat due to the emergence of resistance to synthetic insecticides. Insecticides of plant origin may serve as suitable alternative biocontrol techniques in the future. The purpose of the present study was to assess the ethyl acetate, acetone and methanol extracts of Andrographis paniculata, Eclipta prostrata and Tagetes erecta leaves tested for oviposition-deterrent, ovicidal and repellent activities against malaria vector, Anopheles subpictus Grassi (Diptera: Culicidae). Methods: The dried leaves of the three plants were powdered mechanically and extracted with ethyl acetate, acetone and methanol. One gram of crude extract was first dissolved in 100 ml of acetone (stock solution). From the stock solution, test solution concentrations of 31.21- 499.42 mg/l for oviposition- deterrence assay and repellency and 15.60 - 998.85 mg/l were used in ovicidal assay. The percentage oviposition- deterrence, hatching rate of eggs and protection time were calculated. One-way analysis of variance was used for the multiple concentration tests and for per cent mortality to determine significant treatment differences. Results: The percentage of effective oviposition repellency was highest at 499.42 mg/l and the lowest at 31.21 mg/l in ethyl acetate, acetone and methanol extracts of A. paniculata, E. prostrata and T. erecta. The oviposition activity index (OAI) value of ethyl acetate, acetone and methanol extracts of A. paniculata, E. prostrata and T. erecta at 499.42 mg/l were -0.91, -0.93, -0.84, -0.84, -0.87, -0.82, -0.87, -0.89 and -0.87, respectively. Mortality (no egg hatchability) was 100 per cent with ethyl acetate and methanol extracts of A. paniculata, E. prostrata and T. erecta at 998.85 mg/l. The maximum adult repellent activity was observed at 499.42 mg/l in ethyl acetate extracts of A. paniculata, E. prostrata and methanol extracts of T. erecta, and the mean complete protection time ranged from 120 to 150 min with the different extracts tested. Interpretation & conclusions: The acetone extract of A. paniculata, methanol extract of E. prostrata and T. erecta showed good oviposition-deterrent, ovicidal and repellent activities respectively. These results suggest that the leaf extracts of A. paniculata, E. prostrata and T. erecta may have the potential to be used as an ideal eco-friendly approach for the control of the An. subpictus. PMID:21985822

  4. [PKA-regulated phosphorylation status of S149 and S321 sites of CDC25B inhibits mitosis of fertilized mouse eggs].

    PubMed

    Xiao, Jian-Ying; Liu, Chao; Sun, Xiao-Han; Yu, Bing-Zhi

    2012-02-25

    To further test whether protein kinase A (PKA) can affect the mitotic cell cycle, one-cell stage mouse embryos at S phase (22 h after hCG injection) were incubated in M16 medium containing various concentrations of H-89, a PKA inhibitor. With increasing concentrations of H-89 (0-50 μmol/L), the G(2) phase of eggs was decreased and the cleavage rate was accelerated. A concentration of 40 μmol/L H-89 led to all of the mouse eggs entering the M phase of mitosis. Furthermore, to study the role of PKA in regulating the phosphorylation status of S149 and S321 sites of cell division cycle 25B (CDC25B) on one-cell stage fertilized mouse eggs, pBSK-CDC25B-WT, pBSK-CDC25B-S149A, pBSK-CDC25B-S321A and pBSK-CDC25B-S149A/S321A were transcribed into mRNAs in vitro, then mRNAs were microinjected into S phase of mouse fertilized eggs and cultured in M16 medium pretreated with H-89. Then, the cleavage of fertilized eggs, maturation promoting factor (MPF) activity and phosphorylation status of CDC2-Tyr15 were observed. In the presence of 40 μmol/L H-89, the cleavage rate of fertilized eggs in CDC25B-S/A-mRNAs and CDC25B-WT-mRNA injected groups was significantly higher than that in the control groups, and the peak of MPF activity appeared in the CDC25B-S/A-mRNAs and CDC25B-WT-mRNA injected groups earlier than that in the control groups. CDC2-Tyr15 phosphorylation state was consistent with MPF activity. In conclusion, the present study suggests that PKA regulates the early development of mouse embryos by phosphorylation of S149 and S321 of CDC25B, which plays an important role in the regulation of G(2)/M transition in the mitotic cell cycle of fertilized mouse eggs.

  5. Somatic insulin signaling regulates a germline starvation response in Drosophila egg chambers

    PubMed Central

    Burn, K. Mahala; Shimada, Yuko; Ayers, Kathleen; Lu, Feiyue; Hudson, Andrew M.; Cooley, Lynn

    2014-01-01

    Egg chambers from starved Drosophila females contain large aggregates of processing (P) bodies and cortically enriched microtubules. As this response to starvation is rapidly reversed upon re-feeding females or culturing egg chambers with exogenous bovine insulin, we examined the role of endogenous insulin signaling in mediating the starvation response. We found that systemic Drosophila insulin-like peptides (dILPs) activate the insulin pathway in follicle cells, which then regulate both microtubule and P body organization in the underlying germline cells. This organization is modulated by the motor proteins Dynein and Kinesin. Dynein activity is required for microtubule and P body organization during starvation, while Kinesin activity is required during nutrient-rich conditions. Blocking the ability of egg chambers to form P body aggregates in response to starvation correlated with reduced progeny survival. These data suggest a potential mechanism to maximize fecundity even during periods of poor nutrient availability, by mounting a protective response in immature egg chambers. PMID:25481758

  6. Epithelial rotation promotes the global alignment of contractile actin bundles during Drosophila egg chamber elongation

    PubMed Central

    Cetera, Maureen; Ramirez-San Juan, Guillermina R.; Oakes, Patrick W.; Lewellyn, Lindsay; Fairchild, Michael J.; Tanentzapf, Guy; Gardel, Margaret L.; Horne-Badovinac, Sally

    2014-01-01

    Tissues use numerous mechanisms to change shape during development. The Drosophila egg chamber is an organ-like structure that elongates to form an elliptical egg. During elongation the follicular epithelial cells undergo a collective migration that causes the egg chamber to rotate within its surrounding basement membrane. Rotation coincides with the formation of a “molecular corset”, in which actin bundles in the epithelium and fibrils in the basement membrane are all aligned perpendicular to the elongation axis. Here we show that rotation plays a critical role in building the actin-based component of the corset. Rotation begins shortly after egg chamber formation and requires lamellipodial protrusions at each follicle cell’s leading edge. During early stages, rotation is necessary for tissue-level actin bundle alignment, but it becomes dispensable after the basement membrane is polarized. This work highlights how collective cell migration can be used to build a polarized tissue organization for organ morphogenesis. PMID:25413675

  7. Genetic Variations in Bionomics of Culex quinquefasciatus (Diptera: Culicidae) Mosquito Population in Minna, North Central Nigeria

    PubMed Central

    Ukubuiwe, Azubuike C.; Olayemi, Israel K.; Jibrin, Aisha I.

    2016-01-01

    The need to have an improved knowledge on the bioecology of Culex quinquefasciatus, a prerequisite in the development of cost-effective control strategies, has informed the present preliminary investigation to put in better perspective variations that exist in the egg rafts of the species. Freshly laid egg rafts were collected and incubated at ambient temperature in well-labeled plastic trays. The results showed overall inconsistency in all indices monitored for the egg rafts. Generally, survivorship was high for the species. All immature stage and adult parameters measured varied significantly among the egg rafts and between/within sexes of the species. Therefore, this study suggests the presence of inherent variation in the bionomics of egg rafts of C. quinquefasciatus, probably influenced by the environment and hence underscores the need for additional studies to further elucidate the roles of genetics and environment in vectorial competence of the species, in order to develop robust sustainable mosquito vector control protocols. PMID:27013900

  8. Sexual selection of male parental care in giant water bugs

    PubMed Central

    Ohba, Shin-ya; Okuda, Noboru; Kudo, Shin-ichi

    2016-01-01

    Paternal care can be maintained under sexual selection, if it helps in attracting more mates. We tested the hypothesis in two giant water bug species, Appasus major and Appasus japonicus, that male parental care is sexually selected through female preference for caring males. Females were given an opportunity to choose between two males. In the first test of female mate choice, one male carried eggs on its back, while the other did not. The egg status was switched between these two males in the second test. The experiment revealed that females of both species preferred caring males (i.e. egg-bearing) to non-caring males. Nonetheless, the female mate preference for egg-bearing males was stronger in A. major than in A. japonicus. Our results suggest that sexual selection plays an important role in maintaining elaborate paternal care in giant water bugs, but the importance of egg-bearing by males in female mate choice varies among species. PMID:27293778

  9. Chrysopa septempunctata (Neuroptera: Chrysopidae) Vitellogenin Functions Through Effects on Egg Production and Hatching.

    PubMed

    Liu, C; Mao, J; Zeng, F

    2015-12-01

    Vitellogenin (Vg) is a precursor of major egg storage protein, vitellin (Vt), and plays primary roles in reproduction of oviparous vertebrates and invertebrates. Chrysopa septempunctata Wesmael is an important and common predator of various insect pests. Here, we first cloned C. septempunctata Vg gene, CsVg. The complete CsVg cDNA was 5664 bp, which encodes an 1810-residues protein with a predicted molecular mass of 206.23 kDa. Expression profile revealed that CsVg mRNA first appeared on day 4 after emergence, maximally accumulated on day 10, and then declined gradually. RNAi mediated by injection of dsRNA depleted CsVg transcripts, significantly reduced egg-laying amount, and decreased egg hatching rate, suggesting that CsVg functions through effects on egg production and hatching in C. septempunctata. © The Authors 2015. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  10. Relations of the brown pelican to certain environmental pollutants

    USGS Publications Warehouse

    Blus, L.J.; Belisle, A.A.; Prouty, R.M.

    1974-01-01

    Nearly all brown pelican eggs collected from 13 colonies in South Carolina, Florida, and California in 1969 and from 17 colonies in South Carolina and Florida in 1970 exhibited eggshell thinning. Of the 100 eggs analyzed for residues of pollutants, all eggs contained measurable quantities of DDE; most eggs contained measurable quantities of p,p'-DDD, p,p'-DDT, dieldrin, or PCB's (polychlorinated biphenyls). All eggs contained measurable quantities of mercury. DDE appears to have been responsible for virtually all the eggshell thinning. There is strong evidence that DDE played a major role in lowered reproductive success in South Carolina and California, and this pollutant appears to be intimately related to the population decline in South Carolina. Other pollutants, particularly dieldrin, may have had deleterious effect on reproductive success in South Carolina. Carcasses of pelicans collected by shooting in Florida and South Carolina in 1970 varied in residue load according to age and geographic location. Birds under 1 year of age contained smaller quantities of residues than did birds I year or older.

  11. Transmission of helminth eggs through hands in a high-risk community.

    PubMed

    Gulliver, Francesca; Jeandron, Aurelie; Nguyen, Viet Anh; Do, Hong Anh; Ensink, Jeroen H J

    2014-10-01

    The role that hands play in the transmission of Ascaris is not well understood. A newly developed method to quantify the presence helminth eggs on hands was tested among a group of farmers in Vietnam. High turbidity in hand rinse samples meant that the adopted sugar solution had to be replaced by zinc sulphate as a flotation solution. The survey found 34% (31/90) of study participant hands' were positive for helminth eggs, though concentrations were low and ranged from 0-10 eggs/2 hands. The use of fresh excreta in agriculture was the only variable that showed an association with the concentrations of eggs found on hands. The results highlight the need to promote handwashing with soap to prevent the transmission of soil-transmitted helminth infections. © The Author 2014. Published by Oxford University Press on behalf of Royal Society of Tropical Medicine and Hygiene. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  12. Plant volatiles induced by herbivore egg deposition affect insects of different trophic levels.

    PubMed

    Fatouros, Nina E; Lucas-Barbosa, Dani; Weldegergis, Berhane T; Pashalidou, Foteini G; van Loon, Joop J A; Dicke, Marcel; Harvey, Jeffrey A; Gols, Rieta; Huigens, Martinus E

    2012-01-01

    Plants release volatiles induced by herbivore feeding that may affect the diversity and composition of plant-associated arthropod communities. However, the specificity and role of plant volatiles induced during the early phase of attack, i.e. egg deposition by herbivorous insects, and their consequences on insects of different trophic levels remain poorly explored. In olfactometer and wind tunnel set-ups, we investigated behavioural responses of a specialist cabbage butterfly (Pieris brassicae) and two of its parasitic wasps (Trichogramma brassicae and Cotesia glomerata) to volatiles of a wild crucifer (Brassica nigra) induced by oviposition of the specialist butterfly and an additional generalist moth (Mamestra brassicae). Gravid butterflies were repelled by volatiles from plants induced by cabbage white butterfly eggs, probably as a means of avoiding competition, whereas both parasitic wasp species were attracted. In contrast, volatiles from plants induced by eggs of the generalist moth did neither repel nor attract any of the tested community members. Analysis of the plant's volatile metabolomic profile by gas chromatography-mass spectrometry and the structure of the plant-egg interface by scanning electron microscopy confirmed that the plant responds differently to egg deposition by the two lepidopteran species. Our findings imply that prior to actual feeding damage, egg deposition can induce specific plant responses that significantly influence various members of higher trophic levels.

  13. Nitric oxide synthase during early embryonic development in silkworm Bombyx mori: Gene expression, enzyme activity, and tissue distribution.

    PubMed

    Kitta, Ryo; Kuwamoto, Marina; Yamahama, Yumi; Mase, Keisuke; Sawada, Hiroshi

    2016-12-01

    To elucidate the mechanism for embryonic diapause or the breakdown of diapause in Bombyx mori, we biochemically analyzed nitric oxide synthase (NOS) during the embryogenesis of B. mori. The gene expression and enzyme activity of B. mori NOS (BmNOS) were examined in diapause, non-diapause, and HCl-treated diapause eggs. In the case of HCl-treated diapause eggs, the gene expression and enzyme activity of BmNOS were induced by HCl treatment. However, in the case of diapause and non-diapause eggs during embryogenesis, changes in the BmNOS activity and gene expressions did not coincide except 48-60 h after oviposition in diapause eggs. The results imply that changes in BmNOS activity during the embryogenesis of diapause and non-diapause eggs are regulated not only at the level of transcription but also post-transcription. The distribution and localization of BmNOS were also investigated with an immunohistochemical technique using antibodies against the universal NOS; the localization of BmNOS was observed mainly in the cytoplasm of yolk cells in diapause eggs and HCl-treated diapause eggs. These data suggest that BmNOS has an important role in the early embryonic development of the B. mori. © 2016 Japanese Society of Developmental Biologists.

  14. ["The first stages of the human egg" by Auguste d'Eternod published one hundred years ago in the Comptes Rendus de l'Association des Anatomistes].

    PubMed

    Catala, M

    2014-06-01

    The development of the embryo and foetus fascinates, but its study in humans is difficult because of both technical and ethical problems. Auguste d'Eternod, Swiss embryologist, published in 1913 an article entitled "The early stages of the human egg" in the Comptes Rendus de l'Association des Anatomistes, the ancestor of the journal Morphologie. This work is focused not only on the early stages of development: fertilization, cleavage of the egg, blastocyst formation, gastrulation, but also on the extra-embryonic processes characteristic of mammals. On the occasion of the centenary of the publication of this work, I propose a critical review by placing the data published in the literature and historical context of the time. Finally, I try to extract from these observations the concepts that are still used today by embryologists. Copyright © 2014 Elsevier Masson SAS. All rights reserved.

  15. Biofumigation for control of pale potato cyst nematodes: activity of brassica leaf extracts and green manures on Globodera pallida in vitro and in soil.

    PubMed

    Lord, James S; Lazzeri, Luca; Atkinson, Howard J; Urwin, Peter E

    2011-07-27

    The effects of brassica green manures on Globodera pallida were assessed in vitro and in soil microcosms. Twelve of 22 brassica accessions significantly inhibited the motility of G. pallida infective juveniles in vitro. Green manures of selected brassicas were then incorporated into soil containing encysted eggs of G. pallida. Their effect on egg viability was estimated by quantifying nematode actin 1 mRNA by RT-qPCR. The leaf glucosinolate profiles of the plants were determined by high-performance liquid chromatography. Three Brassica juncea lines (Nemfix, Fumus, and ISCI99) containing high concentrations of 2-propenyl glucosinolate were the most effective, causing over 95% mortality of encysted eggs of G. pallida in polyethylene-covered soil. The toxic effects of green manures were greater in polyethylene-covered than in open soil. Toxicity in soil correlated with the concentration of isothiocyanate-producing glucosinolate but not total glucosinolate in green manures.

  16. How the discovery of ribozymes cast RNA in the roles of both chicken and egg in origin-of-life theories.

    PubMed

    Sankaran, Neeraja

    2012-12-01

    Scientific theories about the origin-of-life theories have historically been characterized by the chicken-and-egg problem of which essential aspect of life was the first to appear, replication or self-sustenance. By the 1950s the question was cast in molecular terms and DNA and proteins had come to represent the carriers of the two functions. Meanwhile, RNA, the other nucleic acid, had played a capricious role in origin theories. Because it contained building blocks very similar to DNA, biologists recognized early that RNA could store information in its linear sequences. With the discovery in the 1980s that RNA molecules were capable of biological catalysis, a function hitherto ascribed to proteins alone, RNA took on the role of the single entity that could act as both chicken and egg. Within a few years of the discovery of these catalytic RNAs (ribozymes) scientists had formulated an RNA World hypothesis that posited an early phase in the evolution of life where all key functions were performed by RNA molecules. This paper traces the history the role of RNA in origin-of-life theories with a focus on how the discovery of ribozymes influenced the discourse. Copyright © 2012 Elsevier Ltd. All rights reserved.

  17. Cytosol-dependent membrane fusion in ER, nuclear envelope and nuclear pore assembly: biological implications.

    PubMed

    Rafikova, Elvira R; Melikov, Kamran; Chernomordik, Leonid V

    2010-01-01

    Endoplasmic reticulum and nuclear envelope rearrangements after mitosis are often studied in the reconstitution system based on Xenopus egg extract. In our recent work we partially replaced the membrane vesicles in the reconstitution mix with protein-free liposomes to explore the relative contributions of cytosolic and transmembrane proteins. Here we discuss our finding that cytosolic proteins mediate fusion between membranes lacking functional transmembrane proteins and the role of membrane fusion in endoplasmic reticulum and nuclear envelope reorganization. Cytosol-dependent liposome fusion has allowed us to restore, without adding transmembrane nucleoporins, functionality of nuclear pores, their spatial distribution and chromatin decondensation in nuclei formed at insufficient amounts of membrane material and characterized by only partial decondensation of chromatin and lack of nuclear transport. Both the mechanisms and the biological implications of the discovered coupling between spatial distribution of nuclear pores, chromatin decondensation and nuclear transport are discussed.

  18. Survey of undeclared egg allergen levels in the most frequently recalled food types (including products bearing precautionary labelling).

    PubMed

    Khuda, Sefat E; Sharma, Girdhari M; Gaines, Dennis; Do, Andrew B; Pereira, Marion; Chang, Michael; Ferguson, Martine; Williams, Kristina M

    2016-08-01

    Since the number of recalls involving undeclared allergens is commonly associated with bakery and snack foods, we aimed to determine the frequency of egg allergens in a large number of these products using two commercial enzyme-linked immunosorbent assay (ELISA) methods. Samples were chosen that either had no egg identified on the product label or which had an egg precautionary statement. Among all samples, egg protein was detected in 5% of products using a Morinaga (MO) kit and 1% of products using a R-Biopharm (RB) kit. For bakery samples, egg protein was detected in 6% of 363 samples with no precautionary labelling (6% by MO and 1% by RB kit) and 12% of 80 samples which had precautionary labelling. For snack samples, egg protein was detected in 2% of 371 samples with no precautionary labelling (2% by MO and < 1% by RB kit) and 5% of 21 samples which had precautionary labelling. The disagreement rates between two methods were 5.2% for bakery products and 2.6% for snack products. The sample repeatability was at an acceptable level for bakery (< 12.5%) and snack foods (< 7.5%) for each method. The relative standard deviation between test kits was high (103.1%) for bakery foods. Four bakery products without precautionary labelling had a higher level of egg protein per serving compared with the eliciting dose (ED10 of 3.7 mg protein) for egg allergic patients. These results highlight the fact that detection methodology plays a vital role for accurate labelling control and mitigation of risk for egg allergic consumers.

  19. The Pathways from a Behavior Change Communication Intervention to Infant and Young Child Feeding in Bangladesh Are Mediated and Potentiated by Maternal Self-Efficacy.

    PubMed

    Zongrone, Amanda A; Menon, Purnima; Pelto, Gretel H; Habicht, Jean-Pierre; Rasmussen, Kathleen M; Constas, Mark A; Vermeylen, Francoise; Khaled, Adiba; Saha, Kuntal K; Stoltzfus, Rebecca J

    2018-02-01

    Although self-efficacy is a potential determinant of feeding and care behaviors, there is limited empirical analysis of the role of maternal self-efficacy in low- and middle-income countries. In the context of behavior change interventions (BCIs) addressing complementary feeding (CF), it is possible that maternal self-efficacy can mediate or enhance intervention impacts. In the context of a BCI in Bangladesh, we studied the role of maternal self-efficacy for CF (MSE-CF) for 2 CF behaviors with the use of a theoretically grounded empirical model of determinants to illustrate the potential roles of MSE-CF. We developed and tested a locally relevant scale for MSE-CF and included it in a survey (n = 457 mothers of children aged 6-24 mo) conducted as part of a cluster-randomized evaluation. Qualitative research was used to inform the selection of 2 intervention-targeted behaviors: feeding green leafy vegetables in the last 24 h (GLV) and on-time introduction of egg (EGG) between 6 and 8 mo of age. We then examined direct, mediated, and potentiated paths of MSE-CF in relation to the impacts of the BCI on these behaviors with the use of regression and structural equation modeling. GLV and EGG were higher in the intensive group than in the nonintensive control group (16.0 percentage points for GLV; P < 0.001; 11.2 percentage points for EGG; P = 0.037). For GLV, MSE-CF mediated (β = 0.345, P = 0.010) and potentiated (β = 0.390, P = 0.038) the effect of the intensive group. In contrast, MSE-CF did not mediate or potentiate the effect of the intervention on EGG. MSE-CF was a significant mediator and potentiator for GLV but not for EGG. The divergent findings highlight the complex determinants of individual specific infant and young child feeding behaviors. The study shows the value of measuring behavioral determinants, such as MSE-CF, that affect a caregiver's capability to adopt intervention-targeted behaviors.

  20. In vivo anthelmintic activity of Dorycnium rectum and grape seed extract against Ostertagia (Teladorsagia) circumcincta and Trichostrongylus colubriformis in sheep.

    PubMed

    Waghorn, T S; Molan, A L; Deighton, M; Alexander, R A; Leathwick, D M; McNabb, W C; Meagher, L P

    2006-02-01

    To assess the in vivo anthelmintic activity of condensed tannins (CT) in the forage species Dorycnium rectum and Medicago sativa, and in an extract from grape (Vitus vinifera) seeds (GSE), against two species of parasite, Teladorsagia (Ostertagia) circumcincta and Trichostrongylus colubriformis, at different stages of their life cycle, in sheep that were parasite-naïve or previously exposed to nematodes. In Trial 1, a factorial treatment structure was used to compare faecal nematode egg counts (FEC) and worm burdens in 40 weaned Romney lambs fed either the CT-containing forage D. rectum (12% dry matter; DM) or M. sativa (lucerne; 0.2% DM). Twenty naïve and 20 previously-exposed lambs were drenched free of parasites then reinfected with known species and numbers of parasites, and housed in pens indoors on a diet of lucerne pellets and chaffed hay. Groups of lambs (n=5 lambs per group) were fed one of the forages over one of two time periods within the parasite's life cycle. Six to nine days after the last feeding of fresh forages, faecal samples were collected for FEC, and all lambs were slaughtered and worm counts conducted. In Trial 2, 12 Suffolk x Romney lambs were surgically implanted with an abomasal cannula and then housed indoors in metabolism crates. After infection with parasites, six lambs were infused continuously over a 14-day period with a commercially available CT GSE (96% DM, made up to 34 g/L in water); the remaining lambs were infused with water. During infusion, samples were collected for egg hatch and larval development assays. After infusion, samples were collected for FEC, and all lambs were slaughtered and worm counts conducted. In Trial 1, there was a significant (p<0.001) difference in burdens of O. circumcincta between naïve lambs and those previously exposed to parasites, but no other differences were recorded. In Trial 2, lambs infused with GSE had significantly (p<0.05) fewer T. colubriformis at slaughter and significantly (p<0.001) fewer eggs hatched in the egg hatch assay (EHA) than for lambs infused with water. Overall, the differences attributable to GSE were small in magnitude, being an 11% drop in egg hatch, and an 18% drop in numbers of adult T. colubriformis after 14 days of continuous infusion. No other differences were recorded. The results indicate that the in vivo anthelmintic activity of these CT sources is, at best, modest and is unlikely to be of any practical value. Further, these data emphasise that in vitro activity is an unreliable indicator of in vivo efficacy for CT-containing forages and extracts.

  1. Scrambled or bisected mouse eggs and the basis of patterning in mammals.

    PubMed

    Gardner, R L

    1999-04-01

    Several findings challenge the notion that specification of cell types and embryonic axes in mammals are rooted entirely in the temporal and spatial relations between cleaving blastomeres. They raise the question as to whether, as in most non-mammalian species, these processes depend on information already present in the egg. However, experiments designed to investigate this possibility directly by perturbing the organization of the zygote or, very recently, by deleting one or other of its polar regions [M. Zernicka-Goetz. Fertile offspring derived from mammalian eggs lacking either animal or vegetal poles. Development 1998;125:4803-4808 (Ref. 1)], have been interpreted to mean that such a role for the egg can be discounted. This conclusion seems premature in view of continuing uncertainty regarding the developmental potential of individual blastomeres in mammals.

  2. A New Cell-Free System to Study BRCA1 Function

    DTIC Science & Technology

    2014-05-01

    antibodies to FANCI, FANCD2, DNA pol e, FANCA , FANCM. We first wanted to test whether Approach, which is inhibited in BRCA1-depleted egg extracts...analysis. We have not initiated this task. 4c. For novel proteins whose binding to chromatin depends on BRCA1: clone the gene , express the protein

  3. Use of a pheromone to cause copulation between two species of cerambycids

    Treesearch

    Jimmy R. Galford

    1980-01-01

    The painted hickory borer, Megacyllene caryae (Gahan) and the locust borer Megacyllene robiniae (Forster) will not attempt to interbreed in the laboratory. However, male locust borers copulated with painted hickory borer females painted with an alcohol extract of female locust borers. No eggs were laid by treated or untreated...

  4. COMPARISON OF SUBCUTANEOUS AND ORAL ROUTES OF EXPOSURE FOR EVALUATING ALLERGENICITY OF FOOD EXTRACTS

    EPA Science Inventory

    Evaluation of the potential for food allergenicity of any given protein is limited by the lack of an appropriate animal model. In this study we examined the intrinsic allergenicity of foods known to be allergenic (peanut, egg) or non-allergenic (spinach) by exposing mice either s...

  5. NSF- and SNARE-mediated membrane fusion is required for nuclear envelope formation and completion of nuclear pore complex assembly in Xenopus laevis egg extracts.

    PubMed

    Baur, Tina; Ramadan, Kristijan; Schlundt, Andreas; Kartenbeck, Jürgen; Meyer, Hemmo H

    2007-08-15

    Despite the progress in understanding nuclear envelope (NE) reformation after mitosis, it has remained unclear what drives the required membrane fusion and how exactly this is coordinated with nuclear pore complex (NPC) assembly. Here, we show that, like other intracellular fusion reactions, NE fusion in Xenopus laevis egg extracts is mediated by SNARE proteins that require activation by NSF. Antibodies against Xenopus NSF, depletion of NSF or the dominant-negative NSF(E329Q) variant specifically inhibited NE formation. Staging experiments further revealed that NSF was required until sealing of the envelope was completed. Moreover, excess exogenous alpha-SNAP that blocks SNARE function prevented membrane fusion and caused accumulation of non-flattened vesicles on the chromatin surface. Under these conditions, the nucleoporins Nup107 and gp210 were fully recruited, whereas assembly of FxFG-repeat-containing nucleoporins was blocked. Together, we define NSF- and SNARE-mediated membrane fusion events as essential steps during NE formation downstream of Nup107 recruitment, and upstream of membrane flattening and completion of NPC assembly.

  6. Determination of chloramphenicol residues in milk, eggs, and tissues by liquid chromatography/mass spectrometry.

    PubMed

    Penney, Lisa; Smith, Anderson; Coates, Brent; Wijewickreme, Arosha

    2005-01-01

    A new liquid chromatography/mass spectrometry (LC/MS) method is presented for the determination of chloramphenicol (CAP) residues in milk, eggs, chicken muscle and liver, and beef muscle and kidney. CAP is extracted from the samples with acetonitrile and defatted with hexane. The acetonitrile extracts are then evaporated, and residues are reconstituted in 10mM ammonium acetate--acetonitrile mobile phase and injected into the LC system. CAP is determined by reversed-phase chromatography using an Inertsil ODS-2 column and MS detection with negative ion electrospray ionization. Calibration curves were linear between 0.5-5.0 ng/g for all matrixes studied. The relative standard deviations for measurements by this method were generally <12%, and average recoveries ranged from 80 to 120%, depending on the matrix involved. The method detection limits of CAP ranged from 0.2 to 0.6 ng/g, which are comparable to previously reported results. The proposed method is rapid, simple, and specific, allowing a single analyst to easily prepare over 40 samples in a regular working day.

  7. The Genetic Basis of Natural Variation in Drosophila (Diptera: Drosophilidae) Virgin Egg Retention.

    PubMed

    Akhund-Zade, Jamilla; Bergland, Alan O; Crowe, Sarah O; Unckless, Robert L

    2017-01-01

    Drosophila melanogaster is able to thrive in harsh northern climates through adaptations in life-history traits and physiological mechanisms that allow for survival through the winter. We examined the genetic basis of natural variation in one such trait, female virgin egg retention, which was previously shown to vary clinally and seasonally. To further our understanding of the genetic basis and evolution of virgin egg retention, we performed a genome-wide association study (GWAS) using the previously sequenced Drosophila Genetic Reference Panel (DGRP) mapping population. We found 29 single nucleotide polymorphisms (SNPs) associated with virgin egg retention and assayed 6 available mutant lines, each harboring a mutation in a candidate gene, for effects on egg retention time. We found that four out of the six mutant lines had defects in egg retention time as compared with the respective controls: mun, T48, Mes-4, and Klp67A Surprisingly, none of these genes has a recognized role in ovulation control, but three of the four genes have known effects on fertility or have high expression in the ovaries. We also found that the SNP set associated with egg retention time was enriched for clinal SNPs. The majority of clinal SNPs had alleles associated with longer egg retention present at higher frequencies in higher latitudes. Our results support previous studies that show higher frequency of long retention times at higher latitude, providing evidence for the adaptive value of virgin egg-retention. © The Author 2017. Published by Oxford University Press on behalf of the Entomological Society of America.

  8. Microbial infections are associated with embryo mortality in Arctic-nesting geese.

    USGS Publications Warehouse

    Hansen, Cristina M.; Meixell, Brandt W.; Van Hemert, Caroline R.; Hare, Rebekah F.; Hueffer, Karsten

    2015-01-01

    To address the role of bacterial infection in hatching failure of wild geese, we monitored embryo development in a breeding population of Greater white-fronted geese (Anser albifrons) on the Arctic Coastal Plain of Alaska. During 2013, we observed mortality of normally developing embryos and collected 36 addled eggs for analysis. We also collected 17 infertile eggs for comparison. Using standard culture methods and gene sequencing to identify bacteria within collected eggs, we identified a potentially novel species of Neisseria in 33 eggs, Macrococcus caseolyticus in 6 eggs, and Streptococcus uberis and Rothia nasimurium in 4 eggs each. We detected seven other bacterial species at lower frequencies. Sequences of the 16S rRNA genes from the Neisseria isolates most closely matched sequences from N. animaloris and N. canis (96 to 97% identity), but phylogenetic analysis suggested substantial genetic differentiation between egg isolates and known Neisseria species. Although definitive sources of the bacteria remain unknown, we detected Neisseria DNA from swabs of eggshells, nest contents, and cloacae of nesting females. To assess the pathogenicity of bacteria identified in contents of addled eggs, we inoculated isolates of Neisseria, Macrococcus, Streptococcus, and Rothia at various concentrations into developing chicken eggs. Seven-day mortality rates varied from 70 to 100%, depending on the bacterial species and inoculation dose. Our results suggest that bacterial infections are a source of embryo mortality in wild geese in the Arctic.    

  9. Laboratory Evaluations of the Fractions Efficacy of Annona senegalensis (Annonaceae) Leaf Extract on Immature Stage Development of Malarial and Filarial Mosquito Vectors

    PubMed Central

    Lame, Younoussa; Nukenine, Elias Nchiwan; Pierre, Danga Yinyang Simon; Elijah, Ajaegbu Eze; Esimone, Charles Okechukwu

    2015-01-01

    Background: Within the framework to control mosquitoes, ovicidal, larvicidal and pupicidal activity of Annona senegalensis leaf extract and its 4 fractions against Anopheles gambiae and Culex quinquefasciatus were evaluated in the laboratory conditions. Methods: Ovicidal test was performed by submitting at least 100 eggs of mosquitoes to 125, 250, 500, 1000 and 2000 ppm concentrations, while larvicidal and pupicidal effects were assessed by submitting 25 larvae or pupae to the concentrations of 2500, 1250, 625 and 312.5 ppm of plant extract or fractions of A. senegalensis. Results: The eggs of An. gambiae were most affected by N-hexane (0.00% hatchability) and chloroform (03.67% hatchability) fractions compared to Cx. quinquefasciatus where at least 25 % hatchability were recorded at 2000 ppm. For larvicidal test, N-hexane (LC50= 298.8 ppm) and chloroform (LC50= 418.3 ppm) fractions were more effective than other fractions on An. gambiae larvae while, a moderate effectiveness was also observed with N-hexane (LC50= 2087.6 ppm), chloroform (LC50= 9010.1 ppm) fractions on Cx. quinquefasciatus larvae. The highest mortality percent of the pupae were also recorded with N-hexane and chloroform fractions on An. gambiae at 2500 ppm. As for Cx. quinquefasciatus only 50 % and 36 % mortality were recorded with N-hexane and chloroform fractions respectively. Conclusion: The extract of A. senegalensis was toxic on immature stage of mosquito species tested. By splitting methanolic crude extract, only N-hexane and chloroform fractions were revealed to possess a mosquitocidal effects and could be considered and utilized for future immature mosquito vectors control. PMID:26623434

  10. Integration: valuing stakeholder input in setting priorities for socially sustainable egg production.

    PubMed

    Swanson, J C; Lee, Y; Thompson, P B; Bawden, R; Mench, J A

    2011-09-01

    Setting directions and goals for animal production systems requires the integration of information achieved through internal and external processes. The importance of stakeholder input in setting goals for sustainable animal production systems should not be overlooked by the agricultural animal industries. Stakeholders play an integral role in setting the course for many aspects of animal production, from influencing consumer preferences to setting public policy. The Socially Sustainable Egg Production Project (SSEP) involved the development of white papers on various aspects of egg production, followed by a stakeholder workshop to help frame the issues for the future of sustainable egg production. Representatives from the environmental, food safety, food retail, consumer, animal welfare, and the general farm and egg production sectors participated with members of the SSEP coordination team in a 1.5-d workshop to explore socially sustainable egg production. This paper reviews the published literature on values integration methodologies and the lessons learned from animal welfare assessment models. The integration method used for the SSEP stakeholder workshop and its outcome are then summarized. The method used for the SSEP stakeholder workshop can be used to obtain stakeholder input on sustainable production in other farm animal industries.

  11. Queen pheromone regulates egg production in a termite.

    PubMed

    Yamamoto, Yuuka; Matsuura, Kenji

    2011-10-23

    In social insects, resource allocation is a key factor that influences colony survival and growth. Optimal allocation to queens and brood is essential for maximum colony productivity, requiring colony members to have information on the total reproductive power in colonies. However, the mechanisms regulating egg production relative to the current labour force for brood care remain poorly known. Recently, a volatile chemical was identified as a termite queen pheromone that inhibits the differentiation of new neotenic reproductives (secondary reproductives developed from nymphs or workers) in Reticulitermes speratus. The same volatile chemical is also emitted by eggs. This queen pheromone would therefore be expected to act as an honest message of the reproductive power about queens. In this study, we examined how the queen pheromone influences the reproductive rate of queens in R. speratus. We compared the number of eggs produced by each queen between groups with and without exposure to artificial queen pheromone. Exposure to the pheromone resulted in a significant decrease in egg production in both single-queen and multiple-queen groups. This is the first report supporting the role of queen pheromones as a signal regulating colony-level egg production, using synthetically derived compounds in a termite.

  12. Queen pheromone regulates egg production in a termite

    PubMed Central

    Yamamoto, Yuuka; Matsuura, Kenji

    2011-01-01

    In social insects, resource allocation is a key factor that influences colony survival and growth. Optimal allocation to queens and brood is essential for maximum colony productivity, requiring colony members to have information on the total reproductive power in colonies. However, the mechanisms regulating egg production relative to the current labour force for brood care remain poorly known. Recently, a volatile chemical was identified as a termite queen pheromone that inhibits the differentiation of new neotenic reproductives (secondary reproductives developed from nymphs or workers) in Reticulitermes speratus. The same volatile chemical is also emitted by eggs. This queen pheromone would therefore be expected to act as an honest message of the reproductive power about queens. In this study, we examined how the queen pheromone influences the reproductive rate of queens in R. speratus. We compared the number of eggs produced by each queen between groups with and without exposure to artificial queen pheromone. Exposure to the pheromone resulted in a significant decrease in egg production in both single-queen and multiple-queen groups. This is the first report supporting the role of queen pheromones as a signal regulating colony-level egg production, using synthetically derived compounds in a termite. PMID:21543395

  13. Role of shell diffusion area in incubating eggs at simulated high altitude.

    PubMed

    Weiss, H S

    1978-10-01

    Embryonic development is inhibited when eggs are incubated at 9,100 m (0.3 atm) despite a normoxic environment. The problem apparently relates to respiratory gas exchange occurring by diffusion through gas-filled pores in the shell. Gaseous flux is therefore inversely proportional to ambient pressure and is affected by the physical characteristics of the ambient gas (Chapman-Enskog equation). Excess loss of H2O and CO2 occurs in eggs incubating at altitude and could be detrimental. Such increased loss should be correctable by decreasing diffusion area. This was tested by progressively increasing coverage of the shell with paraffin and incubating at simulated 0.3 ATA (225 Torr) in 100% O2. Uncoated eggs failed to hatch, but numbers of chicks increased with increased coverage. Maximum hatch was an extrapolated 90% of controls at 69% shell coverage. With further coverage, hatch size decreased. Egg weight loss, and estimate of H2O diffusion, was around three times controls in uncoated eggs but decreased linearly with paraffin coverage, reaching near normal at maximum hatch. Reduction of diffusion area to 0.3 normal at maximum hatch generally balanced the increased flux predicted for 0.3 ATA.

  14. Ability of bed bug-detecting canines to locate live bed bugs and viable bed bug eggs.

    PubMed

    Pfiester, Margie; Koehler, Philip G; Pereira, Roberto M

    2008-08-01

    The bed bug, Cimex lectularius L., like other bed bug species, is difficult to visually locate because it is cryptic. Detector dogs are useful for locating bed bugs because they use olfaction rather than vision. Dogs were trained to detect the bed bug (as few as one adult male or female) and viable bed bug eggs (five, collected 5-6 d after feeding) by using a modified food and verbal reward system. Their efficacy was tested with bed bugs and viable bed bug eggs placed in vented polyvinyl chloride containers. Dogs were able to discriminate bed bugs from Camponotus floridanus Buckley, Blattella germanica (L.), and Reticulitermes flavipes (Kollar), with a 97.5% positive indication rate (correct indication of bed bugs when present) and 0% false positives (incorrect indication of bed bugs when not present). Dogs also were able to discriminate live bed bugs and viable bed bug eggs from dead bed bugs, cast skins, and feces, with a 95% positive indication rate and a 3% false positive rate on bed bug feces. In a controlled experiment in hotel rooms, dogs were 98% accurate in locating live bed bugs. A pseudoscent prepared from pentane extraction of bed bugs was recognized by trained dogs as bed bug scent (100% indication). The pseudoscent could be used to facilitate detector dog training and quality assurance programs. If trained properly, dogs can be used effectively to locate live bed bugs and viable bed bug eggs.

  15. Strategies for the screening of antibiotic residues in eggs: comparison of the validation of the classical microbiological method with an immunobiosensor method.

    PubMed

    Gaudin, Valérie; Rault, Annie; Hedou, Celine; Soumet, Christophe; Verdon, Eric

    2017-09-01

    Efficient screening methods are needed to control antibiotic residues in eggs. A microbiological kit (Explorer® 2.0 test (Zeu Inmunotech, Spain)) and an immunobiosensor kit (Microarray II (AM® II) on Evidence Investigator™ system (Randox, UK)) have been evaluated and validated for screening of antibiotic residues in eggs, according to the European decision EC/2002/657 and to the European guideline for the validation of screening methods. The e-reader™ system, a new automatic incubator/reading system, was coupled to the Explorer 2.0 test. The AM II kit can detect residues of six different families of antibiotics in different matrices including eggs. For both tests, a different liquid/liquid extraction of eggs had to be developed. Specificities of the Explorer 2.0 and AM II kit were equal to 8% and 0% respectively. The detection capabilities were determined for 19 antibiotics, with representatives from different families, for Explorer 2.0 and 12 antibiotics for the AM II kit. For the nine antibiotics having a maximum residue limit (MRL) in eggs, the detection capabilities CCβ of Explorer 2.0 were below the MRL for four antibiotics, equal to the MRL for two antibiotics and between 1 and 1.5 MRLs for the three remaining antibiotics (tetracyclines). For the antibiotics from other families, the detection capabilities were low for beta-lactams and sulfonamides and satisfactory for dihydrostreptomycin (DHS) and fluoroquinolones, which are usually difficult to detect with microbiological tests. The CCβ values of the AM II kit were much lower than the respective MRLs for three detected antibiotics (tetracycline, oxytetracycline, tylosin). Concerning the nine other antibiotics, the detection capabilities determined were low. The highest CCβ was obtained for streptomycin (100 µg kg -1 ).

  16. High-value utilization of egg shell to synthesize Silver and Gold-Silver core shell nanoparticles and their application for the degradation of hazardous dyes from aqueous phase-A green approach.

    PubMed

    Sinha, Tanur; Ahmaruzzaman, M

    2015-09-01

    The common household material, egg shell of Anas platyrhynchos is utilized for the synthesis of Silver and Gold-Silver core shell nanoparticles using greener, environment friendly and economic way. The egg shell extracts were acting as a stabilizing and reducing agents. This method avoids the use of external reducing and stabilizing agents, templates and solvents. The effects of various reaction parameters, such as reaction temperature, concentration in the formation of nanoparticles have also been investigated. The compositional abundance of gelatin may be envisaged for the effective reductive as well as stabilizing potency. The mechanisms for the formation of NPs have also been presented. The synthesized Ag NPs formed were predominantly spherical in nature with an average size of particles in the range of 6-26 nm. While, Au-Ag core shell nanoparticles formed were spherical and oval shaped, within a narrow size spectrum of 9-18 nm. Both the Ag NPs Au-and Ag core shell nanoparticles showed characteristic Bragg's reflection planes of fcc structure and surface plasmon resonance at 430 nm and 365 nm, respectively. The NPs were utilized for the removal of toxic and hazardous dyes, such as Rose Bengal, Methyl Violet 6 B and Methylene Blue from aqueous phase. Approximately 98.2%, 98.4% and 97% degradations of Rose Bengal, Methyl Violet 6 B, and Methylene Blue were observed with Ag NPs, while the percentage degradation of these dyes was 97.3%, 97.6% and 96% with Au-Ag NPs, respectively. Therefore, the present study has opened up an innovative way for synthesizing Ag NPs and Au-Ag bimetallic nanostructures of different morphologies and sizes involving the utilization of egg shell extract. The high efficiency of the NPs as photocatalysts has opened a promising application for the removal of hazardous dyes from the industrial effluents. Copyright © 2015 Elsevier Inc. All rights reserved.

  17. Vertical transmission of microcystins to Nile crocodile (Crocodylus niloticus) eggs.

    PubMed

    Singo, Alukhethi; Myburgh, Jan G; Laver, Peter N; Venter, Elizabeth A; Ferreira, Gezina C H; Rösemann, Gertruida M; Botha, Christo J

    2017-08-01

    Cyanobacteria or blue green algae are known for their extensive and highly visible blooms in eutrophic, stagnant freshwater bodies. Climate change and global warming have also contributed to a rise in toxic cyanobacterial blooms. One of the most important cyanobacteria is Microcystis aeruginosa, which can synthesize various microcystins that can affect the health of terrestrial and aquatic animals. Commercial Nile crocodile (Crocodylus niloticus) farming in South Africa is based on keeping breeders (adult males and females) in big dams on farms (captive-bred approach). Unfortunately, cyanobacterial blooms in the breeder dams are a concern to farm owners, managers and veterinarians. The main objectives of this research project were to determine if microcystins were present in the contents of crocodile eggs and the liver and yolk of dead hatchlings, and to determine if the reduced hatchability on commercial farms might be caused by these toxins. Furthermore, the concentration of microcystins in the breeder dam was monitored on a monthly basis spanning the ovulation and egg laying period. During the hatching season microcystin concentrations in unfertilised eggs, egg shell membranes and in the yolk and liver of dead hatchlings were determined using liquid chromatography-high resolution mass spectrometry (LC-HRMS). Microcystins were detected in Nile crocodile egg and hatchling samples. Microcystin (MC-LR, MC-RR, MC-YR) concentrations in the crocodile egg and hatchling samples collected from clutches with a good hatching rate (≥90%) ranged between 0 and 1.76 ng g -1 , with the highest concentration in the egg shell membranes. Microcystin concentrations in samples collected from clutches with a bad hatching rate (≤10%) ranged from 0 - 1.63 ng g -1 with the highest concentration detected in the hatchling yolk. However, the concentrations were probably underestimated as the percentage recovery from spiked samples was very low with the extraction method employed. Bayesian analysis suggests that the liver, yolk and unfertilised egg all have similar microcystin concentrations, while the membranes have (with moderate to high certainty) higher microcystin concentrations. There appears to be no difference in microcystin concentrations among good and bad clutches across all tissue types or within a specific tissue type, but due to the small sample size, it was not possible to determine whether microcystin affected the hatchability of Nile crocodile eggs. However, vertical transmission of microcystin variants to the Nile crocodile egg does occur and the possible implications for the survival of wild Nile crocodile populations should be ascertained. Copyright © 2017 Elsevier Ltd. All rights reserved.

  18. Involvement of Mos-MEK-MAPK pathway in cytostatic factor (CSF) arrest in eggs of the parthenogenetic insect, Athalia rosae.

    PubMed

    Yamamoto, Daisuke S; Tachibana, Kazunori; Sumitani, Megumi; Lee, Jae Min; Hatakeyama, Masatsugu

    2008-01-01

    Extensive survey of meiotic metaphase II arrest during oocyte maturation in vertebrates revealed that the mitogen-activated protein kinase (MAPK) pathway regulated by the c-mos proto-oncogene product, Mos, has an essential role in cytostatic activity, termed cytostatic factor (CSF). In contrast, little is known in invertebrates in which meiotic arrest occurs in most cases at metaphase I (MI arrest). A parthenogenetic insect, the sawfly Athalia rosae, in which artificial egg activation is practicable, has advantages to investigate the mechanisms of MI arrest. Both the MAPK/extracellular signal-regulated protein kinase kinase (MEK) and MAPK were phosphorylated and maintained active in MI-arrested sawfly eggs, whereas they were dephosphorylated soon after egg activation. Treatment of MI-arrested eggs with U0126, an inhibitor of MEK, resulted in dephosphorylation of MAPK and MI arrest was resumed. The sawfly c-mos gene orthologue encoding a serine/threonine kinase was cloned and analyzed. It was expressed in nurse cells in the ovaries. To examine CSF activity of the sawfly Mos, synthesized glutathione S-transferase (GST)-fusion sawfly Mos protein was injected into MI-resumed eggs in which MEK and MAPK were dephosphorylated. Both MEK and MAPK were phosphorylated again upon injection. In these GST-fusion sawfly Mos-injected eggs subsequent mitotic (syncytial) divisions were blocked and embryonic development was ceased. These results demonstrated that the MEK-MAPK pathway was involved in maintaining CSF arrest in sawfly eggs and Mos functioned as its upstream regulatory molecule.

  19. Influence of extraction pH on the foaming, emulsification, oil-binding and visco-elastic properties of marama protein.

    PubMed

    Gulzar, Muhammad; Taylor, John Rn; Minnaar, Amanda

    2017-11-01

    Marama bean protein, as extracted previously at pH 8, forms a viscous, adhesive and extensible dough. To obtain a protein isolate with optimum functional properties, protein extraction under slightly acidic conditions (pH 6) was investigated. Two-dimensional electrophoresis showed that pH 6 extracted marama protein lacked some basic 11S legumin polypeptides, present in pH 8 extracted protein. However, it additionally contained acidic high molecular weight polypeptides (∼180 kDa), which were disulfide crosslinked into larger proteins. pH 6 extracted marama proteins had similar emulsification properties to soy protein isolate and several times higher foaming capacity than pH 8 extracted protein, egg white and soy protein isolate. pH 6 extracted protein dough was more elastic than pH 8 extracted protein, approaching the elasticity of wheat gluten. Marama protein extracted at pH 6 has excellent food-type functional properties, probably because it lacks some 11S polypeptides but has additional high molecular weight proteins. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

  20. Is the red fox (Vulpes vulpes) a competent definitive host for Taenia multiceps?

    PubMed

    Varcasia, Antonio; Tamponi, Claudia; Tosciri, Gabriele; Pipia, Anna Paola; Dore, Francesco; Schuster, Rolf Karl; Kandil, Omnia Mohamed; Manunta, Maria Lucia; Scala, Antonio

    2015-09-25

    Shepherd and stray dogs are thought to represent the primary definitive hosts of Coenurosis by Taenia multiceps, due to their feeding habits which translate into high chances of coming into contact with infected intermediate hosts. Nonetheless, little attention has been paid to the role of the red fox (Vulpes vulpes) in the epidemiology of coenurosis. In fact a knowledge gap exists on the role played by red foxes in the epidemiology of Taenia multiceps and the capability of this parasite to produce fertile and viable eggs in this wild canid, i.e. on the occurrence of a sylvatic cycle. This study investigates the role of the red fox (Vulpes vulpes) in the epidemiology of T. multiceps and related metacestodoses. The small intestine of 63 red foxes was macroscopically examined for the presence of cestodes. Adult parasites were identified morphologically as being T. multiceps. Tapeworm eggs were counted and stored at 4 °C in physiological saline solution prior to experimental infection of four sheep and one goat. Sheep were inoculated orally on Day 0 with 3000 (sheep 1), 5000 (sheep 2 and 3) or 7000 eggs (sheep 4), while the goat was infected with 5000 eggs of T. multiceps. The animals were followed-up regularly by MRI and underwent surgical treatment between days 180 to day 240 post infection. Collected coenuri were identified using morphological and molecular methods. A total of 6.3 % of red foxes were found infected with T. multiceps and the eggs obtained from the worms were determined to have a viability of 45.4 %. Two of the challenged sheep and the goat developed disease compatible with T. multiceps. Morphometrical features of the cysts were consistent with those of T. multiceps; nucleotide amplification and sequencing of mitochondrial genes (i.e., cox1 and Nd1) from the metacestode material confirmed the identification. The present study is the first to provide evidence of the role of the red fox as a competent definitive host for T. multiceps, thus changing the epidemiological scenarios of infections by this cestode.

  1. Hypersensitive response of beans to Apion godmani (Coleoptera: Curculionidae).

    PubMed

    Garza, R; Vera, J; Cardona, C; Barcenas, N; Singh, S P

    2001-08-01

    High levels of resistance to Apion godinani Wagner have been reported in bean, Phaseolus vulgaris L., landraces from Mexico. We report on the role of hypersensitivity to A. godmani in five resistant and three susceptible bean genotypes. In susceptible genotypes (cultivars 'Canario 107','Jamapa', and 'Zacatecas 45'), the eggs and first instars of A. godmani were embedded in the pod mesocarp and usually were surrounded by healthy tissue. In contrast, in resistant landraces ('Amarillo 154', 'Amarillo 155', 'J-117', 'Puebla 36', and 'Pinto 168'), necrotic tissues developed concentrically around the oviposition site, encapsulating eggs and dead larvae. An inverse relationship between percentage egg and larval encapsulation at the early immature pod stages and percentage of damaged seeds at harvest was found. Results indicate that hypersensitivity in developing pods plays an important role in antibiosis to A. godmani in beans. This information will facilitate future genetic and biochemical research and provide much needed information concerning the phenotypic basis of resistance to A. godmani in bean.

  2. DOE Office of Scientific and Technical Information (OSTI.GOV)

    Santella, Luigia, E-mail: santella@szn.it; Limatola, Nunzia; Chun, Jong T.

    The interaction of the spermatozoon with the egg at fertilization remains one of the most fascinating mysteries of life. Much of our scientific knowledge on fertilization comes from studies on sea urchin and starfish, which provide plenty of gametes. Large and transparent, these eggs have served as excellent model systems for studying egg activation and embryo development in seawater, a plain natural medium. Starfish oocytes allow the study of the cortical, cytoplasmic and nuclear changes during the meiotic maturation process, which can also be triggered in vitro by hormonal stimulation. These morphological and biochemical changes ensure successful fertilization of the eggsmore » at the first metaphase. On the other hand, sea urchin eggs are fertilized after the completion of meiosis, and are particularly suitable for the study of sperm–egg interaction, early events of egg activation, and embryonic development, as a large number of mature eggs can be fertilized synchronously. Starfish and sea urchin eggs undergo abrupt changes in the cytoskeleton and ion fluxes in response to the fertilizing spermatozoon. The plasma membrane and cortex of an egg thus represent “excitable media” that quickly respond to the stimulus with the Ca{sup 2+} swings and structural changes. In this article, we review some of the key findings on the rapid dynamic rearrangements of the actin cytoskeleton in the oocyte/egg cortex upon hormonal or sperm stimulation and their roles in the modulation of the Ca{sup 2+} signals and in the control of monospermic fertilization. - Highlights: • Besides microtubules, microfilaments may anchor the nucleus to oocyte surface. • The cortical Ca{sup 2+} flash and wave at fertilization mirror electrical membrane change. • Artificial egg activation lacks microvilli extension in the perivitelline space. • Calcium is necessary but not sufficient for cortical granules exocytosis. • Actin cytoskeleton modulates Ca{sup 2+} release at oocyte maturation and fertilization.« less

  3. Effects of Xylem-Sap Composition on Glassy-Winged Sharpshooter (Hemiptera: Cicadellidae) Egg Maturation on High- and Low-Quality Host Plants.

    PubMed

    Sisterson, Mark S; Wallis, Christopher M; Stenger, Drake C

    2017-04-01

    Glassy-winged sharpshooters must feed as adults to produce mature eggs. Cowpea and sunflower are both readily accepted by the glassy-winged sharpshooter for feeding, but egg production on sunflower was reported to be lower than egg production on cowpea. To better understand the role of adult diet in egg production, effects of xylem-sap chemistry on glassy-winged sharpshooter egg maturation was compared for females confined to cowpea and sunflower. Females confined to cowpea consumed more xylem-sap than females held on sunflower. In response, females held on cowpea produced more eggs, had heavier bodies, and greater lipid content than females held on sunflower. Analysis of cowpea and sunflower xylem-sap found that 17 of 19 amino acids were more concentrated in cowpea xylem-sap than in sunflower xylem-sap. Thus, decreased consumption of sunflower xylem-sap was likely owing to perceived lower quality, with decreased egg production owing to a combination of decreased feeding and lower return per unit volume of xylem-sap consumed. Examination of pairwise correlation coefficients among amino acids indicated that concentrations of several amino acids within a plant species were correlated. Principal component analyses identified latent variables describing amino acid composition of xylem-sap. For females held on cowpea, egg maturation was affected by test date, volume of excreta produced, and principal components describing amino acid composition of xylem-sap. Principal component analyses aided in identifying amino acids that were positively or negatively associated with egg production, although determining causality with respect to key nutritional requirements for glassy-winged sharpshooter egg production will require additional testing. Published by Oxford University Press on behalf of Entomological Society of America 2017. This work is written by US Government employees and is in the public domain in the US.

  4. Physical and chemical changes in the foreshore of an estuarine beach: Implications for viability and development of horseshoe crab Limulus polyphemus eggs

    USGS Publications Warehouse

    Jackson, N.L.; Smith, D.R.; Nordstrom, K.F.

    2008-01-01

    Knowledge of conditions that favor development of eggs is important for management of species whose population growth is sensitive to early life history survival. Viability and development of the eggs of horseshoe crabs Limulus polyphemus on a sand and gravel beach were evaluated using data gathered on Delaware Bay, USA, from 18 May to 19 June 2004. Eggs were transplanted to pouches and buried in the foreshore for up to 6 wk. Viability and developmental stage were estimated as a function of oxygen and temperature gradients across the foreshore. These gradients were related to the characteristics of the intertidal foreshore sediments, beach water table changes, and frequency of inundation due to tide and swash/backwash processes. Results demonstrate the importance of interstitial temperature for development to larvae and the passive role of sediment characteristics on moisture retention and temperature. Percentage of eggs remaining in egg stage was similar across the foreshore, but more eggs developed to embryos at 0.45 of foreshore width, where moisture and gravel content were greater and interstitial temperature was lower. More eggs developed to larvae at 0.60 and 0.75 of foreshore width, where moisture and gravel content were less but interstitial temperature was higher. The beach above 0.75 of foreshore width came under the influence of wave action or full tidal inundation only during high wave heights or spring tides, and pouches at 0.75 of foreshore width were inundated only 19% of the time. Periodic wetting at this elevation did not reduce overall viability of the eggs. High wave energy events resulted in sediment activation depths to pouches at 0.30 of foreshore width, where loss of eggs due to wave activation was the most important control on the development of eggs. ?? Inter-Research 2008.

  5. Localization and expression of peroxiredoxin II in the mouse ovary, oviduct, uterus, and preimplantation embryo.

    PubMed

    Wang, Shie; Huang, Weiquan; Shi, Hexiu; Lin, Cuiying; Xie, Meirong; Wang, Jianxin

    2010-02-01

    Peroxiredoxin (Prx) II belongs to a recently discovered family of peroxidases that play important roles in antioxidation and signal transduction. In this study, we aimed to study the localization and expression of Prx II in the mouse ovary, oviduct, and uterus, and preimplantation embryos. Immunohistochemical staining analysis showed that, in the ovary, Prx II was expressed in the oocyte cytoplasm of the primary follicle, the secondary follicle, and the premature follicle; Prx II was expressed in germinal vesicle-intact oocytes (GV oocytes) and metaphase II eggs (MII eggs), as well as at various stages in early embryos. Reverse transcription polymerase chain reaction (RT-PCR) results indicated that the Prx II mRNA was expressed at a high level in GV eggs, slightly lower levels in MII eggs, and had no detectable expression in four-cell embryos and early blastocysts. In the oviduct, Prx II was expressed in the epithelia, while in the uterus Prx II was mainly distributed in the endometrial stroma. Taken together, our results suggest that Prx II plays a key antioxidation role in the maturation of oocytes and development of early embryos, thus providing crucial experimental evidence for further exploring the function of Prx II in the development of oocytes and preimplantation embryos. 2009 Wiley-Liss, Inc.

  6. In vitro evidence for the participation of Drosophila melanogaster sperm β-N-acetylglucosaminidases in the interactions with glycans carrying terminal N-acetylglucosamine residues on the egg's envelopes.

    PubMed

    Intra, Jari; Veltri, Concetta; De Caro, Daniela; Perotti, Maria Elisa; Pasini, Maria Enrica

    2017-09-01

    Fertilization is a complex and multiphasic process, consisting of several steps, where egg-coating envelope's glycoproteins and sperm surface receptors play a critical role. Sperm-associated β-N-acetylglucosaminidases, also known as hexosaminidases, have been identified in a variety of organisms. Previously, two isoforms of hexosaminidases, named here DmHEXA and DmHEXB, were found as intrinsic proteins in the sperm plasma membrane of Drosophila melanogaster. In the present work, we carried out different approaches using solid-phase assays in order to analyze the oligosaccharide recognition ability of D. melanogaster sperm hexosaminidases to interact with well-defined carbohydrate chains that might functionally mimic egg glycoconjugates. Our results showed that Drosophila hexosaminidases prefer glycans carrying terminal β-N-acetylglucosamine, but not core β-N-acetylglucosamine residues. The capacity of sperm β-N-acetylhexosaminidases to bind micropylar chorion and vitelline envelope was examined in vitro assays. Binding was completely blocked when β-N-acetylhexosaminidases were preincubated with the glycoproteins ovalbumin and transferrin, and the monosaccharide β-N-acetylglucosamine. Overall, these data support the hypothesis of the potential role of these glycosidases in sperm-egg interactions in Drosophila. © 2017 Wiley Periodicals, Inc.

  7. Characterization of Tolypocladium cylindrosporum (Hypocreales: Ophiocordycipitaceae) and Its Impact Against Aedes aegypti and Aedes albopictus Eggs at Low Temperature.

    PubMed

    Flor-Weiler, Lina B; Rooney, Alejandro P; Behle, Robert W; Muturi, Ephantus J

    2017-09-01

    We examined the growth characteristics of Tolypocladium cylindrosporum IBT 41712 and its potential to infect eggs of Aedes aegypti and Ae. albopictus at a low temperature (15°C). When grown on Sabouraud dextrose agar supplemented with yeast extract, the IBT 41712 formed white colonies turning to a slightly darker, off-white color when mature. The mycelia bore swollen conidiophores producing smooth-walled, oblong to cylindrical conidia with varying sizes, ranging from 1.5 to 3.5 μm long. To determine the optimum temperature for the fungus, we cultured the fungus at eight temperatures (4°C, 12°C, 15°C, 21°C, 28°C, 33°C, 37°C, and 40°C) and measured the diametric growth. The optimum temperature for growth was 28°C since it had the highest diametric growth rate (2.1 ± 0.05 mm/day) and the fastest sporulation period (within 8-10 days of incubation). There was no fungal growth at the 3 highest temperatures (33°C, 37°C, and 40°C) but plates incubated at 33°C, when shifted to optimal temperature (28°C), showed visible growth indicating that following incubation at 33°C, the fungus remained viable. The IBT 41712 successfully infected mosquito eggs at 15°C. Fungal treatment induced egg hatch on moist seed-germination paper and this effect was more pronounced in Ae. aegypti compared to Ae. albopictus. When treated eggs were immersed in dH 2 O 21 days posttreatment, larval hatch of both Ae. aegypti (control = 91%, 1 × 10 7 conidia/ml, fungal treatment = 0%) and Ae. albopictus (control = 85%, fungal treatment = 28%) was significantly lower in fungal treatment compared to the controls. The ability of the strain to grow in a wide temperature range, and effectively infect mosquito eggs and induce egg hatch at a low temperature warrants further investigation for its potential as a mosquito control agent targeting eggs that overwinter or undergo long diapause.

  8. Role of egg predation by haddock in the decline of an Atlantic herring population

    PubMed Central

    Richardson, David E.; Hare, Jonathan A.; Fogarty, Michael J.; Link, Jason S.

    2011-01-01

    Theoretical studies suggest that the abrupt and substantial changes in the productivity of some fisheries species may be explained by predation-driven alternate stable states in their population levels. With this hypothesis, an increase in fishing or a natural perturbation can drive a population from an upper to a lower stable-equilibrium population level. After fishing is reduced or the perturbation ended, this low population level can persist due to the regulatory effect of the predator. Although established in theoretical studies, there is limited empirical support for predation-driven alternate stable states in exploited marine fish populations. We present evidence that egg predation by haddock (Melanogrammus aeglefinus) can cause alternate stable population levels in Georges Bank Atlantic herring (Clupea harengus). Egg predation by haddock explains a substantial decoupling of herring spawning stock biomass (an index of egg production) from observed larval herring abundance (an index of egg hatching). Estimated egg survival rates ranged from <2–70% from 1971 to 2005. A population model incorporating egg predation and herring fishing explains the major population trends of Georges Bank herring over four decades and predicts that, when the haddock population is high, seemingly conservative levels of fishing can still precipitate a severe decline in the herring population. These findings illustrate how efforts to rebuild fisheries can be undermined by not incorporating ecological interactions into fisheries models and management plans. PMID:21825166

  9. Plant Volatiles Induced by Herbivore Egg Deposition Affect Insects of Different Trophic Levels

    PubMed Central

    Fatouros, Nina E.; Lucas-Barbosa, Dani; Weldegergis, Berhane T.; Pashalidou, Foteini G.; van Loon, Joop J. A.; Dicke, Marcel; Harvey, Jeffrey A.; Gols, Rieta; Huigens, Martinus E.

    2012-01-01

    Plants release volatiles induced by herbivore feeding that may affect the diversity and composition of plant-associated arthropod communities. However, the specificity and role of plant volatiles induced during the early phase of attack, i.e. egg deposition by herbivorous insects, and their consequences on insects of different trophic levels remain poorly explored. In olfactometer and wind tunnel set-ups, we investigated behavioural responses of a specialist cabbage butterfly (Pieris brassicae) and two of its parasitic wasps (Trichogramma brassicae and Cotesia glomerata) to volatiles of a wild crucifer (Brassica nigra) induced by oviposition of the specialist butterfly and an additional generalist moth (Mamestra brassicae). Gravid butterflies were repelled by volatiles from plants induced by cabbage white butterfly eggs, probably as a means of avoiding competition, whereas both parasitic wasp species were attracted. In contrast, volatiles from plants induced by eggs of the generalist moth did neither repel nor attract any of the tested community members. Analysis of the plant’s volatile metabolomic profile by gas chromatography-mass spectrometry and the structure of the plant-egg interface by scanning electron microscopy confirmed that the plant responds differently to egg deposition by the two lepidopteran species. Our findings imply that prior to actual feeding damage, egg deposition can induce specific plant responses that significantly influence various members of higher trophic levels. PMID:22912893

  10. Mouse SLLP1, a sperm lysozyme-like protein involved in sperm-egg binding and fertilization.

    PubMed

    Herrero, María Belén; Mandal, Arabinda; Digilio, Laura C; Coonrod, Scott A; Maier, Bernhard; Herr, John C

    2005-08-01

    This study demonstrates the retention of mouse sperm lysozyme-like protein (mSLLP1) in the equatorial segment of spermatozoa following the acrosome reaction and a role for mSLLP1 in sperm-egg binding and fertilization. Treatment of cumulus intact oocytes with either recmSLLP1 or its antiserum resulted in a significant (P < or = 0.05) inhibition of fertilization. Co-incubation of zona-free mouse oocytes with capacitated mouse spermatozoa in the presence of varying concentrations of anti-recmSLLP1 serum or recmSLLP1 also inhibited sperm-oolemma binding. A complete inhibition of binding and fusion of spermatozoa to the oocyte occurred at 12.5 muM concentration of recmSLLP1, while conventional chicken and human lysozymes did not block sperm-egg binding. mSLLP1 showed receptor sites in the perivitelline space as well as on the microvillar region of the egg plasma membrane. The retention of mSLLP1 in the equatorial segment of acrosome-reacted sperm, the inhibitory effects of both recmSLLP1 and antibodies to SLLP1 on in vitro fertilization with both cumulus intact and zona-free eggs, and the definition of complementary SLLP1-binding sites on the egg plasma membrane together support the hypothesis that a c lysozyme-like protein is involved in the binding of spermatozoa to the egg plasma membrane during fertilization.

  11. Invariant natural killer T cells from children with versus without food allergy exhibit differential responsiveness to milk-derived sphingomyelin.

    PubMed

    Jyonouchi, Soma; Abraham, Valsamma; Orange, Jordan S; Spergel, Jonathan M; Gober, Laura; Dudek, Emily; Saltzman, Rushani; Nichols, Kim E; Cianferoni, Antonella

    2011-07-01

    A key immunologic feature of food allergy (FA) is the presence of a T(h)2-type cytokine bias. Ligation of the invariant natural killer T cell (iNKT) T-cell receptor (TCR) by sphingolipids presented via the CD1d molecule leads to copious secretion of T(h)2-type cytokines. Major food allergens (eg, milk, egg) are the richest dietary source of sphingolipids (food-derived sphingolipids [food-SLs]). Nonetheless, the role of iNKTs in FA is unknown. To investigate the role of iNKTs in FA and to assess whether food-SL-CD1d complexes can engage the iNKT-TCR and induce iNKT functions. PBMCs from 15 children with cow's milk allergy (MA), 12 children tolerant to cow's milk but with allergy to egg, and 13 healthy controls were incubated with α-galactosylceramide (αGal), cow's milk-sphingomyelin, or hen's egg-ceramide. iNKTs were quantified, and their cytokine production and proliferation were assessed. Human CD1d tetramers loaded with milk-sphingomyelin or egg-ceramide were used to determine food-SL binding to the iNKT-TCR. Milk-sphingomyelin, but not egg-ceramide, can engage the iNKT-TCR and induce iNKT proliferation and T(h)2-type cytokine secretion. Children with FA, especially those with MA, had significantly fewer peripheral blood iNKTs and their iNKTs exhibited a greater T(h)2 response to αGal and milk-sphingomyelin than iNKTs of healthy controls. iNKTs from children with FA, especially those with MA, are reduced in number and exhibit a T(h)2 bias in response to αGal and milk-sphingomyelin. These data suggest a potential role for iNKTs in FA. Copyright © 2011 American Academy of Allergy, Asthma & Immunology. Published by Mosby, Inc. All rights reserved.

  12. Expressed microRNA associated with high rate of egg production in chicken ovarian follicles.

    PubMed

    Wu, N; Gaur, U; Zhu, Q; Chen, B; Xu, Z; Zhao, X; Yang, M; Li, D

    2017-04-01

    MicroRNA (miRNA) is a highly conserved class of small noncoding RNA about 19-24 nucleotides in length that function in a specific manner to post-transcriptionally regulate gene expression in organisms. Tissue miRNA expression studies have discovered a myriad of functions for miRNAs in various aspects, but a role for miRNAs in chicken ovarian tissue at 300 days of age has not hitherto been reported. In this study, we performed the first miRNA analysis of ovarian tissues in chickens with low and high rates of egg production using high-throughput sequencing. By comparing low rate of egg production chickens with high rate of egg production chickens, 17 significantly differentially expressed miRNAs were found (P < 0.05), including 11 known and six novel miRNAs. We found that all 11 known miRNAs were involved mainly in pathways of reproduction regulation, such as steroid hormone biosynthesis and dopaminergic synapse. Additionally, expression profiling of six randomly selected differentially regulated miRNAs were validated by quantitative real-time polymerase chain reaction (RT-qPCR). Some miRNAs, such as gga-miR-34b, gga-miR-34c and gga-miR-216b, were reported to regulate processes such as proliferation, cell cycle, apoptosis and metastasis and were expressed differentially in ovaries of chickens with high rates of egg production, suggesting that these miRNAs have an important role in ovary development and reproductive management of chicken. Furthermore, we uncovered that a significantly up-regulated miRNA-gga-miR-200a-3p-is ubiquitous in reproduction-regulation-related pathways. This miRNA may play a special central role in the reproductive management of chicken, and needs to be further studied for confirmation. © 2016 Stichting International Foundation for Animal Genetics.

  13. [Determination of proximal chemical composition of squid (dosidicus gigas) and development of gel products].

    PubMed

    Abugoch, L; Guarda, A; Pérez, L M; Paredes, M P

    1999-06-01

    The good nutritional properties of meat from big squid (Dosidicus gigas) living on the Chilean coast, was determined through its proximal composition 70 cal/100 g fresh meat; 82.23 +/- 0.98% moisture; 15.32 +/- 0.93% protein; 1.31 +/- 0.12% ashes; 0.87 +/- 0.18% fat and 0.27% NNE (non-nitrogen extract). The big squid meat was used to develop a gel product which contained NaCl and TPP. It was necessary to use additives for gel preparation, such as carragenin or alginate or egg albumin, due to the lack of gelation properties of squid meat. Formulations containing egg albumin showed the highest gel force measured by penetration as compared to those that contained carragenin or alginate.

  14. Anthelmintic activity of Cocos nucifera L. against sheep gastrointestinal nematodes.

    PubMed

    Oliveira, L M B; Bevilaqua, C M L; Costa, C T C; Macedo, I T F; Barros, R S; Rodrigues, A C M; Camurça-Vasconcelos, A L F; Morais, S M; Lima, Y C; Vieira, L S; Navarro, A M C

    2009-01-22

    The development of anthelmintic resistance has made the search for alternatives to control gastrointestinal nematodes of small ruminants imperative. Among these alternatives are several medicinal plants traditionally used as anthelmintics. This work evaluated the efficacy of Cocos nucifera fruit on sheep gastrointestinal parasites. The ethyl acetate extract obtained from the liquid of green coconut husk fiber (LGCHF) was submitted to in vitro and in vivo tests. The in vitro assay was based on egg hatching (EHT) and larval development tests (LDT) with Haemonchus contortus. The concentrations tested in the EHT were 0.31, 0.62, 1.25, 2.5 and 5 mg ml(-1), while in the LDT they were 5, 10, 20, 40 and 80 mg ml(-1). The in vivo assay was a controlled test. In this experiment, 18 sheep infected with gastrointestinal nematodes were divided into three groups (n=6), with the following doses administered: G1-400 mg kg(-1) LGCHF ethyl acetate extract, G2-0.2 mg kg(-1) moxidectin (Cydectin) and G3-3% DMSO. The worm burden was analyzed. The results of the in vitro and in vivo tests were submitted to ANOVA and analyzed by the Tukey and Kruskal-Wallis tests, respectively. The extract efficacy in the EHT and LDT, at the highest concentrations tested, was 100% on egg hatching and 99.77% on larval development. The parameters evaluated in the controlled test were not statistically different, showing that despite the significant results of the in vitro tests, the LGCHF ethyl acetate extract showed no activity against sheep gastrointestinal nematodes.

  15. Anthelmintic effect of Psidium guajava and Tagetes erecta on wild-type and Levamisole-resistant Caenorhabditis elegans strains.

    PubMed

    Piña-Vázquez, Denia M; Mayoral-Peña, Zyanya; Gómez-Sánchez, Maricela; Salazar-Olivo, Luis A; Arellano-Carbajal, Fausto

    2017-04-18

    Psidium guajava and Tagetes erecta have been used traditionally to treat gastrointestinal parasites, but their active metabolites and mechanisms of action remain largely unknown. To evaluate the anthelmintic potential of Psidium guajava and Tagetes erecta extracts on Levamisole-sensitive and Levamisole-resistant strains of the model nematode Caenorhabditis elegans. Aqueous extracts of Psidium guajava (PGE) and Tagetes erecta (TEE) were assayed on locomotion and egg-laying behaviors of the wild-type (N2) and Levamisole-resistant (CB193) strains of Caenorhabditis elegans. Both extracts paralyzed wild-type and Levamisole-resistant nematodes in a dose-dependent manner. In wild-type worms, TEE 25mg/mL induced a 75% paralysis after 8h of treatment and PGE 25mg/mL induced a 100% paralysis after 4h of treatment. PGE exerted a similar paralyzing effect on N2 wild-type and CB193 Levamisole-resistant worms, while TEE only partially paralyzed CB193 worms. TEE 25mg/mL decreased N2 egg-laying by 65% with respect to the untreated control, while PGE did it by 40%. Psidium guajava leaves and Tagetes erecta flower-heads possess hydrosoluble compounds that block the motility of Caenorhabditis elegans by a mechanism different to that of the anthelmintic drug Levamisole. Effects are also observable on oviposition, which was diminished in the wild-type worms. The strong anthelmintic effects in crude extracts of these plants warrants future work to identify their active compounds and to elucidate their molecular mechanisms of action. Copyright © 2017 Elsevier Ireland Ltd. All rights reserved.

  16. Effects of sexual maturation and Salmonella infection on the expression of Toll-like receptors in the chicken vagina.

    PubMed

    Michailidis, G; Theodoridis, A; Avdi, M

    2011-02-01

    Toll-like receptors (TLRs) are a critical component of the innate immune response in many vertebrates, including avian species. The recent findings of chicken TLRs (cTLRs) expression in ovarian follicles and in the chicken ovary in vivo, as well as the changes in their expression in response to lipopolysaccharide or Salmonella enteritidis (SE) infection, have broad implications for reproductive physiology and for the prevention of transmission of zoonotic diseases to humans through the consumption of contaminated poultry eggs. Because the main route of egg contamination is from infection of the oviduct and mainly from the vagina, the aim of this study was to investigate the expression of the ten cTLRs identified to date in the chicken oviduct in vivo, to determine whether sexual maturation affects their mRNA abundance and to investigate whether SE infection alters the expression of TLRs in the chicken vagina. RNA was extracted from the vagina of healthy prepubertal, sexually mature and aged birds, and from sexually mature and aged SE infected birds. RT-PCR analysis revealed that all types of cTLRs apart from TLR1-1 were expressed in the vagina of sexually mature birds. Quantitative real-time PCR analysis revealed that the mRNA abundance of TLR2-1, 2-2 and 4 differ with respect to sexual maturation in the chicken vagina. SE infection resulted in a significant induction of TLR5 and 15 in the vagina of sexually mature birds, and in a significant induction of TLR2-1, 4 and 15 in the vagina of aged birds, while a significant down-regulation was observed for TLR7 in the vagina of sexually mature birds. These findings suggest that a TLR mediated immune response mechanism exists in the chicken vagina, playing a crucial role in preventing microbial pathogens from being incorporated into newly forming eggs. Copyright © 2011 Elsevier B.V. All rights reserved.

  17. [Immunochemical study of the proteins of various tissues in Crustacea (Decapoda): nature, role, origin].

    PubMed

    Durliat, M

    1983-11-01

    The main proteins of the haemolymph of Crustacea Decapoda have been identified and analysed: haemocyanin, plasma coagulogen, heteroagglutinins, vitellogenins, and molt-related proteins. All these complex components exhibit a high molecular weight and as oligomeric fractions are able to aggregate or dissociate in subunits according to the composition of medium and experimental procedures. Besides their important rôle in the defense mechanism, some proteins are involved in the edification of diverse tissues. They are detected within different compartments: soft integument, calcified carapace and hepatopancreas. They are either in transit or sequestered or synthetized within these tissues. In the crayfish Astacus leptodactylus, some components have been identified in different compartments: --in aqueous extracts from soft integument: the haemocyanin, coagulogen and both fraction F1 (lipoprotein with an approximate molecular weight of 45 kdal) and fraction F2 related to the molt. Both coagulogen and fraction F2 appear sometimes as melanized. These two latter fractions exhibit some glucose-mannose residues and they occur with a higher relative amount than in the blood. --in soluble extracts from calcified cuticle: among the numerous fractions showing a high molecular weight, the haemocyanin and coagulogen are detected. --in aqueous extracts from hepatopancreas: both haemocyanin and coagulogen appear with a little relative amount. Components termed as Fa and Fb are found with a high concentration. One minor fraction is also detected. --in aqueous extracts from eggs: the haemocyanin and fraction Fb are present. Other proteins showing only some antigenic identities with those of the haemolymph are also detected in all these tissues. The haemolymph proteins are not present within these compartments following a passive diffusion. Indeed, their relative amount varies according to the tissue investigated and is different from that found in the blood. Except the haemocyanin detected in all tissues with different aggregation states, the haemolymph proteins identified vary in the organs studied. A qualitative and quantitative selection occurs when the blood proteins enter the other compartments. Perhaps some other proteins are not detected following alterations underwent either in the epithelial barriers or during the tannage process or the chitino-proteic complex formation or due to experimental procedures. On the other hand, each tissue has its own proteins. The integument contains crustacyanins alpha, beta, gamma; the eggs are mainly constituted of lipovitellins and the hepatopancreas is rich in small molecular weight proteins and digestive enzymes.(ABSTRACT TRUNCATED AT 400 WORDS)

  18. Sequence-Specific Interaction between the Disintegrin Domain of Mouse ADAM 3 and Murine Eggs: Role of β1 Integrin-associated Proteins CD9, CD81, and CD98

    PubMed Central

    Takahashi, Yuji; Bigler, Dora; Ito, Yasuhiko; White, Judith M.

    2001-01-01

    ADAM 3 is a sperm surface glycoprotein that has been implicated in sperm-egg adhesion. Because little is known about the adhesive activity of ADAMs, we investigated the interaction of ADAM 3 disintegrin domains, made in bacteria and in insect cells, with murine eggs. Both recombinant proteins inhibited sperm-egg binding and fusion with potencies similar to that which we recently reported for the ADAM 2 disintegrin domain. Alanine scanning mutagenesis revealed a critical importance for the glutamine at position 7 of the disintegrin loop. Fluorescent beads coated with the ADAM 3 disintegrin domain bound to the egg surface. Bead binding was inhibited by an authentic, but not by a scrambled, peptide analog of the disintegrin loop. Bead binding was also inhibited by the function-blocking anti-α6 monoclonal antibody (mAb) GoH3, but not by a nonfunction blocking anti-α6 mAb, or by mAbs against either the αv or β3 integrin subunits. We also present evidence that in addition to the tetraspanin CD9, two other β1-integrin-associated proteins, the tetraspanin CD81 as well as the single pass transmembrane protein CD98 are expressed on murine eggs. Antibodies to CD9 and CD98 inhibited in vitro fertilization and binding of the ADAM 3 disintegrin domain. Our findings are discussed in terms of the involvement of multiple sperm ADAMs and multiple egg β1 integrin-associated proteins in sperm-egg binding and fusion. We propose that an egg surface “tetraspan web” facilitates fertilization and that it may do so by fostering ADAM–integrin interactions. PMID:11294888

  19. Sequence-specific interaction between the disintegrin domain of mouse ADAM 3 and murine eggs: role of beta1 integrin-associated proteins CD9, CD81, and CD98.

    PubMed

    Takahashi, Y; Bigler, D; Ito, Y; White, J M

    2001-04-01

    ADAM 3 is a sperm surface glycoprotein that has been implicated in sperm-egg adhesion. Because little is known about the adhesive activity of ADAMs, we investigated the interaction of ADAM 3 disintegrin domains, made in bacteria and in insect cells, with murine eggs. Both recombinant proteins inhibited sperm-egg binding and fusion with potencies similar to that which we recently reported for the ADAM 2 disintegrin domain. Alanine scanning mutagenesis revealed a critical importance for the glutamine at position 7 of the disintegrin loop. Fluorescent beads coated with the ADAM 3 disintegrin domain bound to the egg surface. Bead binding was inhibited by an authentic, but not by a scrambled, peptide analog of the disintegrin loop. Bead binding was also inhibited by the function-blocking anti-alpha6 monoclonal antibody (mAb) GoH3, but not by a nonfunction blocking anti-alpha6 mAb, or by mAbs against either the alphav or beta3 integrin subunits. We also present evidence that in addition to the tetraspanin CD9, two other beta1-integrin-associated proteins, the tetraspanin CD81 as well as the single pass transmembrane protein CD98 are expressed on murine eggs. Antibodies to CD9 and CD98 inhibited in vitro fertilization and binding of the ADAM 3 disintegrin domain. Our findings are discussed in terms of the involvement of multiple sperm ADAMs and multiple egg beta1 integrin-associated proteins in sperm-egg binding and fusion. We propose that an egg surface "tetraspan web" facilitates fertilization and that it may do so by fostering ADAM-integrin interactions.

  20. Injury-induced rapid activation of MAPK signaling in dechorionated eggs and larvae of the silkworm Bombyx mori.

    PubMed

    Gu, Shi-Hong; Chen, Chien-Hung

    2017-04-01

    Previous study showed that diapause in Bombyx mori eggs can be terminated by dechorionation and that activation in the mitogen-activated protein kinase (MAPK)/extracellular signal-regulated kinase (ERK) in dechorionated cultured eggs is involved in diapause termination. In the present study, the possible mechanism underlying activation of ERK upon dechorionation was further investigated. Results showed that mechanical injury of diapause eggs without medium incubation also resulted in rapid increase in the phospho-ERK levels and that injury increased the phospho-ERK levels at different stages of both diapause eggs and eggs in which diapause initiation was prevented by HCl. Effects of anaerobiosis on dechorionation-stimulated phospho-ERK levels showed that the mechanical injury itself but not the dramatic increase in oxygen uptake upon injury is involved in a rapid activation of ERK. Chemical anaerobiosis on dechorionation-stimulated phospho-ERK levels and the in vivo effect of anaerobiosis showed that the supply of oxygen also plays a role in ERK signaling. In addition, injury induced the phosphorylation of c-jun N-terminal kinases (JNKs) and p38 kinase, components of two parallel MAPK pathways. A kinase assay showed a dramatic increase in JNK kinase activity in egg lysates upon injury. When newly hatched first instar larvae were injured, an increase in the phospho-ERK levels similar to that in dechorionated eggs was observed. From the results, we hypothesize that the injury-induced rapid activation of MAPK signaling, which serves as a natural signal for embryonic development, is related to diapause termination in dechorionated eggs. © 2015 Institute of Zoology, Chinese Academy of Sciences.

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