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Sample records for rana pipiens populations

  1. Effective population sizes and temporal stability of genetic structure in Rana pipiens, the northern leopard frog.

    PubMed

    Hoffman, Eric A; Schueler, Frederick W; Blouin, Michael S

    2004-11-01

    Although studies of population genetic structure are very common, whether genetic structure is stable over time has been assessed for very few taxa. The question of stability over time is particularly interesting for frogs because it is not clear to what extent frogs exist in dynamic metapopulations with frequent extinction and recolonization, or in stable patches at equilibrium between drift and gene flow. In this study we collected tissue samples from the same five populations of leopard frogs, Rana pipiens, over a 22-30 year time interval (11-15 generations). Genetic structure among the populations was very stable, suggesting that these populations were not undergoing frequent extinction and colonization. We also estimated the effective size of each population from the change in allele frequencies over time. There exist few estimates of effective size for frog populations, but the data available suggest that ranid frogs may have much larger ratios of effective size (Ne) to census size (Nc) than toads (bufonidae). Our results indicate that R. pipiens populations have effective sizes on the order of hundreds to at most a few thousand frogs, and Ne/Nc ratios in the range of 0.1-1.0. These estimates of Ne/Nc are consistent with those estimated for other Rana species. Finally, we compared the results of three temporal methods for estimating Ne. Moment and pseudolikelihood methods that assume a closed population gave the most similar point estimates, although the moment estimates were consistently two to four times larger. Wang and Whitlock's new method that jointly estimates Ne and the rate of immigration into a population (m) gave much smaller estimates of Ne and implausibly large estimates of m. This method requires knowing allele frequencies in the source of immigrants, but was thought to be insensitive to inexact estimates. In our case the method may have failed because we did not know the true source of immigrants for each population. The method may be more

  2. Variations in the expressed antimicrobial peptide repertoire of northern leopard frog (Rana pipiens) populations suggest intraspecies differences in resistance to pathogens

    PubMed Central

    Tennessen, Jacob A.; Woodhams, Douglas C.; Chaurand, Pierre; Reinert, Laura K.; Billheimer, Dean; Shyr, Yu; Caprioli, Richard M.; Blouin, Michael S.; Rollins-Smith, Louise A.

    2010-01-01

    The northern leopard frog (Rana pipiens or Lithobates pipiens) is historically found in most of the provinces of Canada and the northern and southwest states of the United States. In the last 50 years, populations have suffered significant losses, especially in the western regions of the species range. Using a peptidomics approach, we show that the pattern of expressed antimicrobial skin peptides of frogs from three geographically separated populations are distinct, and we report the presence of four peptides (brevinin-1Pg, brevinin-1Pl, ranatuerin-2Pb, and ranatuerin-2Pc) that have not previously been found in skin secretions. The differences in expressed peptides reflect differences in the distribution of alleles for the newly described Brevinin1.1 locus in the three populations. When enriched peptide mixtures were tested for their ability to inhibit growth of the pathogenic amphibian chytrid (Batrachochytrium dendrobatidis), peptides from Minnesota or Vermont frogs were more effective that peptides from Michigan frogs. Four of the purified peptides were tested for their ability to inhibit growth of two bacterial pathogens (Aeromonas hydrophila and Staphylococcus epidermidis) and B. dendrobatidis. Three of the four were effective inhibitors of B. dendrobatidis and S. epidermidis, but none inhibited A. hydrophila. We interpret these differences in expression and activity of antimicrobial peptides as evidence to suggest that each population may have been selected to express a suite of peptides that reflects current and past encounters with skin microbes. PMID:19622371

  3. Mitotic activity in dorsal epidermis of Rana pipiens.

    NASA Technical Reports Server (NTRS)

    Garcia-Arce, H.; Mizell, S.

    1972-01-01

    Study of statistically significant rhythms of mitotic division in dorsal epidermis of frogs, Rana pipiens, exposed to a 12:12 light:dark environment for 14 days. The results include the findings that (1) male animals have a primary period of 22 hr in summer and 18 hr in winter, (2) female animals have an 18 hr period, and (3) parapinealectomy and blinding abolish the rhythm.

  4. Photoinduced toxicity of fluoranthene to northern leopard frogs (Rana pipiens)

    SciTech Connect

    Monson, P.D.; Call, D.J.; Cox, D.A.; Liber, K.; Ankley, G.T.

    1999-02-01

    Rana pipiens larvae were exposed for 48 h in a flow-through system to clean water or five concentrations of the phototoxic polycyclic aromatic hydrocarbon (PAH) fluoranthene. Following this uptake period, the larvae were divided into four groups: one for immediate tissue residue analysis, a second for residue analysis following 48 h of depuration in clean water, and two for a 48-h exposure in clean water to ultraviolet (UV) light at two different levels. At the highest treatment, mean intensity was 8.12 {+-} 0.19 {times} 10{sup 2} {micro}W/cm{sup 2}, whereas at a lower treatment the UVA intensity was 4.45 {+-} 0.05 {times} 10{sup 2} {micro}W/cm{sup 2}. Larval frogs bioaccumulated fluoranthene in direct proportion to the water exposure concentrations, with initial whole-body PAH concentrations of 1.48, 3.53, 4.85, 11.3, and 18.7 {micro}g/g at the five treatment levels. No mortality of the animals occurred during the 48-h uptake phase. When the frogs were placed in clean water, the fluoranthene was rapidly depurated, with up to 80% lost in 48 h. Exposure to UV light following fluoranthene exposure significantly enhanced toxicity of the PAH. Median time to death decreased as the product of UVA light intensity and fluoranthene body residue increased. For larval R. Pipiens, sufficient tissue residues of fluoranthene were bioaccumulated within 48 h, at water exposure concentrations in the range of 2 to 10 {micro}g/L, to be lethal when combined with a UVA exposure simulating a fraction of summertime, midday sunlight in northern latitudes.

  5. Directionality of phase locking in auditory nerve fibers of the leopard frog Rana pipiens pipiens.

    PubMed

    Schmitz, B; White, T D; Narins, P M

    1992-06-01

    A dorsal approach to the eighth nerve and free-field stimulation were used to investigate the effect of sound direction and intensity on phase locking in auditory nerve fibers of the leopard frog Rana pipiens pipiens. Tuning curves of 75 auditory neurons were analyzed (Fig. 2). Amphibian papillar neurons, but not basilar papillar neurons, exhibit significant phase locking to short tone bursts at the characteristic frequency (CF), the degree of phase locking (vector strength) decreasing with the neuron's CF (Figs. 3, 4 and 10E). Vector strength increases with sound pressure level to saturate about 20 dB above threshold, while the preferred firing phase is only slightly affected (Figs. 5 and 6). In contrast, sound direction hardly affects vector strength (Figs. 7, 8, 9A and 10A and C), but has a strong influence on the preferred firing phase (Figs. 7, 8, 9B and C, 10B and D): With respect to anterior tone presentation there are phase lags for ipsilateral and phase leads for posterior and contralateral presentation. Phase differences between both ears show a sinusoidal or cardioid/ovoidal directional characteristic; maximum differences are found with antero-lateral tone presentation (Fig. 11). The directionality of phase locking decreases with the neuron's CF (Fig. 10F) and only slightly changes with sound pressure level (Fig. 12). Thus, phase locking of amphibian papilla neurons can potentially provide intensity-independent information for sound localization. PMID:1507157

  6. Effects of exposure to ultraviolet light on the development of Rana pipiens, the northern leopard frog

    SciTech Connect

    Williams, J.J.; Wofford, H.W.

    1996-10-01

    The increase in ultraviolet light intensity levels due to ozone depletion recently has been linked to the decline in amphibian population. In this experiment, eggs and larvae of Rana pipiens were subjected to differing amounts of ultraviolet radiation to determine the effects of ultraviolet light on the development of amphibian tadpoles. The total length, length of body without tail, and maximum width of each specimen was recorded for a month of the tadpoles` development, including several measurements after the ultraviolet exposures were concluded. It was found that ultraviolet exposure significantly reduced the size of the organisms in comparison with the control group in all three measured areas. Ultraviolet radiation altered the health and appearance of the exposed organisms and was lethal at large amounts. This experiment showed that ultraviolet radiation could cause many problems in developing amphibians. By slowing their development and physically weakening predation, thus contributing to a decline in overall population levels.

  7. The Developmental Effects Of A Municipal Wastewater Effluent On The Northern Leopard Frog, Rana pipiens

    EPA Science Inventory

    Wastewater effluents are complex mixtures containing a variety of anthropogenic compounds, many of which are known endocrine disruptors. In order to characterize the development and behavorial effects of such a complex mixture, northern leopard frogs, Rana pipiens, were e...

  8. Phylogenetic relationships of leopard frogs (Rana pipiens complex) from an isolated coastal mountain range in southern Sonora, Mexico.

    PubMed

    Pfeiler, E; Markow, T A

    2008-10-01

    Mitochondrial DNA sequence data from the control region and 12S rRNA in leopard frogs from the Sierra El Aguaje of southern Sonora, Mexico, together with GenBank sequences, were used to infer taxonomic identity and provide phylogenetic hypotheses for relationships with other members of the Rana pipiens complex. We show that frogs from the Sierra El Aguaje belong to the Rana berlandieri subgroup, or Scurrilirana clade, of the R. pipiens group, and are most closely related to Rana magnaocularis from Nayarit, Mexico. We also provide further evidence that Rana magnaocularis and R. yavapaiensis are close relatives. PMID:18625327

  9. Distribution and postbreeding environmental relationships of Northern leopard frogs (Rana [Lithobates] pipiens) in Washington

    USGS Publications Warehouse

    Germaine, S.S.; Hays, D.W.

    2009-01-01

    Northern leopard frogs (Rana [Lithobates] pipiens) are considered sensitive, threatened, or endangered in all western states and western Canadian provinces. Historically present in eastern Washington in 6 major river drainages, leopard frogs are now only known to occur at 2 localized areas in the Crab Creek drainage in Grant County. During the summers of 2002-2005, we surveyed both areas to document extent of leopard frog distributions and to describe habitat and vertebrate community characteristics associated with leopard frog site occupancy. At Gloyd Seeps, 2 juvenile leopard frogs were observed in a total of 8.2 person-days of searching along a 5-km stream reach. At Potholes Reservoir, we surveyed 243 wetland sites in 7 management units known to have been occupied by leopard frogs during the 1980s. We confirmed leopard frog presence at only 87 sites (36%) in 4 management units. Site occupancy models for individual ponds indicated that, compared to unoccupied sites, occupied sites had slightly greater pond depths, less tall emergent vegetation, more herbaceous vegetative cover, and fewer neighboring ponds containing nonnative predatory fish. Models developed at the 1-km2 scale indicated that occupied areas had greater average midsummer pond depths, fewer ponds occupied by bullfrogs (Rana [Lithobates] catesbeiana) and carp (Cyprinus carpio), and more herbaceous vegetation surrounding ponds. The Gloyd Seeps population now appears defunct, and the Potholes Reservoir population is in sharp decline. Unless management actions are taken to reduce nonnative fish and bullfrogs and to enhance wetland vegetation, leopard frogs may soon be extirpated from both sites and possibly, therefore, from Washington.

  10. Temperature-dependent elaboration of collagenase by the renal adenocarcinoma of the leopard frog, Rana pipiens.

    PubMed

    Ogilvie, D J; McKinnell, R G; Tarin, D

    1984-08-01

    Naturally occurring renal adenocarcinoma in North American leopard frogs, Rana pipiens, metastasize frequently (77%) when these ectothermic animals are kept in a warm environment but not when they are kept cold. We have found that explants of these tumors secrete collagenase, an enzyme capable of dissolving connective tissue fibers and found previously to be closely correlated with metastatic colony-forming capability of murine mammary tumors, and that the amount released sequentially rises and falls as the ambient temperature is shifted between metastasis-permissive and -inhibitory levels. In contrast, normal frog renal tissue has low collagenase output, unaffected by temperature changes. PMID:6331646

  11. Effects of wetland vs. landscape variables on parasite communities of Rana pipiens: links to anthropogenic factors

    USGS Publications Warehouse

    Schotthoefer, Anna M.; Rohr, Jason R.; Cole, Rebecca A.; Koehler, Anson V.; Johnson, Catherine M.; Johnson, Lucinda B.; Beasley, Val R.

    2011-01-01

    The emergence of several diseases affecting amphibian populations worldwide has prompted investigations into determinants of the occurrence and abundance of parasites in frogs. To understand the spatial scales and identify specific environmental factors that determine risks of parasitism in frogs, helminth communities in metamorphic frogs of the northern leopard frog (Rana pipiens) were examined in relation to wetland and landscape factors at local (1 km) and regional (10 km) spatial extents in an agricultural region of Minnesota (USA) using regression analyses, ordination, and variance partitioning techniques. Greater amounts of forested and woody wetland habitats, shorter distances between woody wetlands, and smaller-sized open water patches in surrounding landscapes were the most consistently positive correlates with the abundances, richness, and diversity of helminths found in the frogs. Wetland and local landscape variables were suggested as most important for larval trematode abundances, whereas local and regional landscape variables appeared most important for adult helminths. As previously reported, the sum concentration of atrazine and its metabolite desethylatrazine, was the strongest predictor of larval trematode communities. In this report, we highlight the additional influences of landscape factors. In particular, our data suggest that anthropogenic activities that have resulted in the loss of the availability and connectivity of suitable habitats in the surrounding landscapes of wetlands are associated with declines in helminth richness and abundance, but that alteration of wetland water quality through eutrophication or pesticide contamination may facilitate the transmission of certain parasite taxa when they are present at wetlands. Although additional research is needed to quantify the negative effects of parasitism on frog populations, efforts to reduce inputs of agrochemicals into wetlands to limit larval trematode infections may be warranted

  12. Effects of polychlorinated biphenyl 126 on green frog (Rana clamitans) and leopard frog (Rana pipiens) hatching success, development, and metamorphosis

    SciTech Connect

    Rosenshield, M.L.; Jofre, M.B.; Karasov, W.H.

    1999-11-01

    Although increasing evidence links plana chlorinated hydrocarbons, such as polychlorinated biphenyls (PCBs), to decreases in survival and reproduction of fish, mammals, and birds near Green Bay, Wisconsin, and the Great Lakes, USA, relatively little is known of their bioaccumulation or of their possible effects in amphibians. The authors exposed embryos and larvae of two ranid species commonly occurring in the Green Bay ecosystem, the green frog (Rana clamitans) and the leopard frog (Rana pipiens), to PCB 126, a model coplanar PCB compound. Nominal concentrations ranged from 0.005 to 50 {micro}g/L, and exposure lasted through metamorphosis. Tissue concentrations of PCB 126 in tadpoles that did not metamorphose by the end of the experiment ranged from 1.2 to 9,600 ng/g wet mass. No significant mortality of embryos occurred before hatching; however, survival of larvae was significantly reduced at the highest concentration for both species. Few deformities were observed, but the incidence of edema was significantly higher in tadpoles exposed to 50 {micro}g/L. Swimming speed and growth of tadpoles was also significantly reduced in this treatment. The percent of tadpoles that reached metamorphosis was significantly lower in green frogs at the highest concentration, and no leopard frogs survived past day 47 of the experiment in this treatment. At high concentrations, PCB 126 affected both ranid species; however, sublethal effects were not apparent for the parameters the authors measured at concentrations that occur in water in the Green Bay ecosystem.

  13. Humane anesthesia and pain management in amphibian limb surgery of Rana pipiens.

    PubMed

    Koustubhan, Punita; Kaplan, David L; Levin, Michael

    2013-02-01

    Adult Rana pipiens frogs are used as a model to investigate mechanisms of vertebrate organ regeneration, anti-tumor ribonucleases, zoological impacts of various pollutants, oncogenesis, neuroplasticity, and neurogenesis. In regenerative biology, the adult Rana pipiens frog is an important alternative to other frog models, such as Xenopus laevis, because it offers the opportunity to study and attempt to augment limb regeneration in an animal that spends significant time out of water and bears weight on its limbs. To elucidate regenerative processes, it is necessary to amputate the limb to study the processes associated with wound healing, blastema formation, and morphogenesis. Being able to revive the animal successfully with little or no side effects is paramount to these studies. Anesthesia and the effect it has on the frogs can vary based on the methods and post-operative care exercised during surgery. However, useful information is not readily available regarding current anesthesia methods or effective and humane analgesia use in amphibians. Amphibian patients are very sensitive to drug dosages, changes in temperature, humidity and water quality; here, special attention is given to these factors. This protocol establishes a humane anesthesia technique while maintaining physiological homeostasis during procedures in amphibians as well as a post-operative care plan addressing the clinical benefits of using analgesics in pain management. Suggestions for infection prevention are covered with a sample treatment plan to ensure that all of the animals have a positive outcome and all of the surgeries have reproducible results. PMID:23378649

  14. Chloride conductance in amphibian skin: regulatory control in the skin of Rana pipiens.

    PubMed

    Rozman, A; Katz, U; Nagel, W

    2008-09-01

    Chloride conductance across the isolated skin of Rana pipiens shows a voltage-activated component (G(Cl)(V)) which requires the presence of mucosal Cl. G(Cl)(V) is normally low or dormant. It is stimulated by elevated intracellular cAMP, irrespective whether originating from application of ss-adrenergic agonists (isoproterenol), stimulators of the adenylyl-cyclase (forskolin), inhibitors of the phosphodiesterases (isobutyl-methyl-xanthine) or membrane-permeable cAMP analogues (CPT-cAMP). Baseline G(Cl) under inactivating conditions increases also with cAMP dose-dependently. The data indicate that cAMP is a central regulator of the passive, conductive chloride transport across amphibian skin. PMID:18599332

  15. Classification and distribution of large intestinal bacteria in nonhibernating and hibernating leopard frogs (Rana pipiens).

    PubMed Central

    Banas, J A; Loesche, W J; Nace, G W

    1988-01-01

    The large intestinal flora of the leopard frog, Rana pipiens, was examined to determine whether differences existed between the nonhibernating and hibernating states of the animal and to determine the relative concentrations and proportions of potential frog pathogens. Hibernators had a logarithmic decrease of bacteria per milligram of intestine averaging one, and significantly greater proportions of facultative bacteria and psychrophiles relative to nonhibernators. The predominant anaerobic bacteria were gram-positive Clostridium species and gram-negative Bacteroides and Fusobacterium species. The predominant facultative bacteria were enterobacteria in nonhibernators but Pseudomonas species in hibernators. Many species of Pseudomonas are pathogenic for frogs, and thus the intestinal flora in hibernators may be a potential source of infectious disease. PMID:3263838

  16. Metamorphic changes in localization of sugars in skin of the leopard frog, Rana pipiens.

    PubMed

    Faszewski, Ellen E; Tyrell, Annemarie; Guin, Sujata; Kaltenbach, Jane C

    2008-08-01

    A lectin histochemical study was carried out to determine the distribution of specific sugars in glycoconjugates within an important osmoregulatory organ, amphibian skin. Paraffin sections were made of Rana pipiens skin from dorsal and ventral regions of aquatic larvae in representative developmental stages as well as from several body regions of semiaquatic adult frogs. Sections were incubated with horseradish peroxidase (HRP)-conjugated lectins, which bind to specific terminal sugar residues of glycoconjugates. Such sites were visualized by DAB-H2O2. The following HRP-lectins were used: UEA-1 for alpha-L-fucose, SBA for N-acetyl-D-galactosamine, WGA for N-acetyl-beta-D-glucosamine, PNA for beta-galactose, and Con A for alpha-mannose. We found that lectin binding patterns in larvae change during metamorphic climax as the skin undergoes extensive histological remodeling; this results in adult skin with staining patterns that are specific for each lectin and are similar in all body regions. Such findings in R. pipiens provide additional insight into the localization of molecules involved in osmoregulation in amphibian skin. PMID:18512706

  17. Characterization and steroidal regulation of gonadotropin beta subunits in the male leopard frog, Rana pipiens.

    PubMed

    Zhang, Lihong; Kessler, Ann E; Tsai, Pei-San

    2007-01-01

    In ranid frogs, the secretion of gonadotropins (GtHs), luteinizing hormone (LH), and follicle-stimulating hormone (FSH), is potently regulated by gonadal steroids. To better understand the gonadal regulation of GtHs at the molecular level, we elucidated the full-length cDNA sequences of LH and FSH beta subunits from the leopard frog, Rana pipiens. The cDNAs for LHbeta and FSHbeta were 1084 and 667 bp in size excluding the poly (A) tail, and encoded proteins of 138 and 127 amino acids, respectively. Using reverse-transcription polymerase chain reaction (RT-PCR), the messages for LHbeta and FSHbeta were found in the pituitary, but not in the brain, heart, kidney, or the liver. Semi-quantitative RT-PCR revealed a significant elevation of FSHbeta, but not LHbeta, in mature male R. pipiens 21 days after gonadectomy (GDX). 17beta-estradiol implant for 21 days in GDX male frogs significantly suppressed the levels of both LHbeta and FSHbeta transcripts, whereas 5alpha-dihydrotestosterone implant suppressed only the latter. Together, these results laid the groundwork for investigating gonadal regulation of GtHbeta subunits in a ranid frog. Importantly, these data also revealed differential feedback effects of an androgen and an estrogen upon GtHbeta expression. PMID:16920113

  18. Influence of Ribeiroia ondatrae (Trematoda: Digenea) infection on limb development and survival of northern leopard frogs (Rana pipiens): effects of host stage and parasite-exposure level

    USGS Publications Warehouse

    Schotthoefer, Anna M.; Koehler, Anson V.; Meteyer, Carol U.; Cole, Rebecca A.

    2003-01-01

    Recent evidence suggests that infection by larvae of the trematode Ribeiroia ondatrae accounts for a significant proportion of limb malformations currently observed in amphibian populations of North America. However, the effects of R. ondatrae infection on northern leopard frogs (Rana pipiens), one of the species most frequently reported with malformations, have not been adequately explored. Moreover, the risk factors associated with R. ondatrae-induced malformations have not been clearly identified. We examined the effects of timing of infection on tadpole survival and limb development. Rana pipiens tadpoles were individually exposed to R. ondatrae cercariae at the pre-limb-bud (Gosner stages 24 and 25), limb-bud (Gosner stages 27 and 28), or paddle (Gosner stages 31–33) stages of development and monitored through metamorphosis. The effects of infection were stage-specific. Infections acquired at the pre-limb-bud stage resulted in a high mortality rate (47.5–97.5%), whereas tadpoles infected at the limb-bud stage displayed a high malformation rate (16% overall), and the magnitude of effects increased with the level of exposure to cercariae. In contrast, infections acquired at the paddle stage had no effect on limb development or tadpole survival, which suggests that the timing of R. ondatrae infection in relation to the stage structure of tadpole populations in the wild is an important determinant of the degree to which populations are affected by R. ondatrae.

  19. Evolution of the functional properties of pyruvate kinase isozymes: pyruvate kinase L from Rana pipiens.

    PubMed

    Fournier, P; Guderley, H

    1986-01-01

    The regulatory properties of type L pyruvate kinase from Rana pipiens are intermediate between those of the mammalian K and L isozymes. As with mammalian type L, the levels of the frog isozyme are affected by the animal's nutritional state. The mammalian and amphibian isozymes show similar sensitivities to fructose 1,6-bisphosphate activation and amino acid inhibition. By contrast, the frog L isozyme shares several properties of the K class: i.e. irreversible inactivation by oxidized glutathione and lack of response to a cyclic AMP stimulated phosphorylation. Furthermore, as for some mammalian K isozymes, frog type L shows a high PEP affinity and a low cooperativity of PEP binding. Insofar as the properties of this present day enzyme reflect those of its counterpart in the amphibian ancestor of higher vertebrates, our results suggest that at its first expression, the type L resembled the type K. Many important regulatory properties of the L isozyme, especially the sensitivity to phosphorylation, were acquired more recently perhaps in association with an increased importance of constant blood glucose. PMID:3489743

  20. Oxidative stress induced in PCB 126-exposed northern leopard frogs, Rana pipiens

    USGS Publications Warehouse

    Huang, Y.-W.; Hoffman, D.J.; Karasov, W.H.

    2007-01-01

    Northern leopard frogs Rana pipiens exposed to PCB 126 (3,3',4,4',5-pentachlorobiphenyl) were examined for hepatic oxidative stress. In a dose-response study, northern leopard frogs were injected intraperitoneally with either PCB 126 in corn oil (0.2, 0.7, 2.3, or 7.8 mg/kg body weight) or corn oil alone. In a time-course study, frogs received 7.8 mg/kg or corn oil alone, and were examined at 1, 2, 3, and 4 wk after dosing. Hepatic concentrations of reduced glutathione (GSH), thiobarbituric acid-reactive substances (TBARS), and total sulfhydryls (total SH), as well as activities of glutathione peroxidase (GSH-P), GSSG reductase (GSSG-R), glucose-6-phosphate dehydrogenase (G-6-PDH), and glutathione S-transferase (GSH-S-T) were measured. In the dose-response experiment, few effects were apparent 1 wk after dosing. In the time-course experiment, significant changes were observed in the 7.8-mg/kg group at 2 wk or more posttreatment. Hepatic concentrations of GSH and TBARS were higher than in corresponding controls at wk 3 and 4; the activities of GSSG-R and GSH-S-T were higher than in controls at wk 2 and 4; and the activity of G-6-PDH was increased at wk 2 and 4. These data collectively indicate that altered glutathione metabolism and oxidative stress occurred and were indicative of both toxicity and induction of protective mechanisms in frogs exposed to PCB. A similar delay in response was reported in fish and may relate to lower metabolic rate and physiological reactions in ectothermic vertebrates

  1. Population Genetic and Admixture Analyses of Culex pipiens Complex (Diptera: Culicidae) Populations in California, United States

    PubMed Central

    Kothera, Linda; Nelms, Brittany M.; Reisen, William K.; Savage, Harry M.

    2013-01-01

    Microsatellite markers were used to genetically characterize 19 Culex pipiens complex populations from California. Two populations showed characteristics of earlier genetic bottlenecks. The overall FST value and a neighbor-joining tree suggested moderate amounts of genetic differentiation. Analyses using Structure indicated K = 4 genetic clusters: Cx. pipiens form pipiens L., Cx. quinquefasciatus Say, Cx. pipiens form molestus Forskäl, and a group of genetically similar individuals of hybrid origin. A Discriminant Analysis of Principal Components indicated that the latter group is a mixture of the other three taxa, with form pipiens and form molestus contributing somewhat more ancestry than Cx. quinquefasciatus. Characterization of 56 morphologically autogenous individuals classified most as Cx. pipiens form molestus, and none as Cx. pipiens form pipiens or Cx. quinquefasciatus. Comparison of California microsatellite data with those of Cx. pipiens pallens Coquillett from Japan indicated the latter does not contribute significantly to genotypes in California. PMID:23958909

  2. Pesticide distributions and population declines of California alpine frogs, Rana muscosa and Rana sierrae

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Atmospherically deposited pesticides from the intensively cultivated Central Valley of California have been implicated as a cause for population declines of several amphibian species, with the strongest evidence for the frogs, Rana muscosa and Rana sierrae at high elevation in the Sierra Nevada moun...

  3. Pesticide Distributions and Population Declines of California Alpine Frogs, Rana Muscosa and Rana Sierrae

    EPA Science Inventory

    Atmospherically deposited pesticides from the intensively cultivated Central Valley of California have been implicated as a cause for population declines of several amphibian species, with the strongest evidence for the frogs Rana muscosa and Rana sierrae at high elevation in th...

  4. Bioaccumulation kinetics of the conventional energetics TNT and RDX relative to insensitive munitions constituents DNAN and NTO in Rana pipiens tadpoles.

    PubMed

    Lotufo, Guilherme R; Biedenbach, James M; Sims, Jerre G; Chappell, Pornsawan; Stanley, Jacob K; Gust, Kurt A

    2015-04-01

    The manufacturing of explosives and their loading, assembling, and packing into munitions for use in testing on training sites or battlefields has resulted in contamination of terrestrial and aquatic sites that may pose risk to populations of sensitive species. The bioaccumulative potential of the conventional explosives 2,4,6-trinitrotoluene (TNT) and hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) and of the insensitive munitions (i.e., less shock sensitive) compound 2,4-dinitroanisole (DNAN) were assessed using the Northern leopard frog, Rana pipiens. Trinitrotoluene entering the organism was readily biotransformed to aminodinitrotoluenes, whereas no transformation products were measured for RDX or DNAN. Uptake clearance rates were relatively slow and similar among compounds (1.32-2.19 L kg(-1) h(-1) ). Upon transfer to uncontaminated water, elimination rate was very fast, resulting in the prediction of fast time to approach steady state (5 h or less) and short elimination half-lives (1.2 h or less). A preliminary bioconcentration factor of 0.25 L kg(-1) was determined for the insensitive munitions compound 3-nitro-1,2,4-trizole-5-one (NTO) indicating negligible bioaccumulative potential. Because of the rapid elimination rate for explosives, tadpoles inhabiting contaminated areas are expected to experience harmful effects only if under constant exposure conditions given that body burdens can rapidly depurate preventing tissue concentrations from persisting at levels that may cause detrimental biological effects. PMID:25524181

  5. Characterization of [3H]-diprenorphine binding in Rana pipiens: observations of filter binding enhanced by naltrexone.

    PubMed

    Newman, L C; Wallace, D R; Stevens, C W

    1999-02-01

    Initial studies were undertaken to examine the properties of [3H]-diprenorphine binding to Rana pipiens whole brain tissue using naltrexone for the definition of nonspecific binding. Saturation analysis demonstrated the binding of [3H]-diprenorphine to be saturable with a K(D) value of 0.65 nM and a Bmax value of 287.7 fmol/mg protein. Unlabeled diprenorphine dose-dependently displaced [3H]-diprenorphine from a single noninteractive site in competition studies which yielded a Ki of 0.22 nM. However, control studies in the absence of tissue revealed significant binding of [3H]-diprenorphine to the filter alone. Interestingly, [3H]-diprenorphine in the presence of unlabeled naltrexone as well as with unlabeled naloxone showed significantly greater binding to the filter than did [3H]-diprenorphine alone. Given this observation of increased nonspecific binding, an artificially low Bmax value would be expected. It is our hypothesis that the unlabeled nonspecific drug forms a complex with [3H]-diprenorphine preventing it from being effectively washed through the filter or the unlabeled drug itself is blocking the flow of [3H]-diprenorphine through the filter. The latter is unlikely however as other binding studies done in our lab using the radioligand [3H]-naloxone with unlabeled naltrexone do not show significant binding to the filter. PMID:10507757

  6. The involvement of the endothelium in the relaxation of the leopard frog (Rana pipiens) aorta in response to acetylcholine.

    PubMed Central

    Knight, G. E.; Burnstock, G.

    1996-01-01

    1. The vasodilator response to acetylcholine (ACh) was investigated in the aortic arches of the leopard frog (Rana pipiens). 2. With adrenaline pre-constricted preparations, both ACh and sodium nitroprusside (SNP) caused concentration-dependent relaxations. Damage to the endothelial layer abolished relaxations to ACh, or reduced them greatly, but had no effect on vasodilatation to SNP. 3. NG-Nitro-L-arginine methyl ester (L-NAME; 1-100 microM) concentration-dependently inhibited relaxations in response to ACh, but had no effect on the ability of SNP to induce vasodilatation. 4. L-Arginine (L-Arg; 100-200 times the concentration of L-NAME) failed to reverse the inhibitory effect of L-NAME (1-100 microM) apart from one isolated instance. 5. In summary, this study has shown endothelium-dependent vasodilatation to ACh in an amphibian blood vessel that appears to be mediated via nitric oxide (NO). The response to ACh differs from many mammalian preparations in that the inhibitory effect of L-NAME could not be overcome by L-Arg, in addition to L-NAME itself having no direct effect upon the tone of the vessel. PMID:8832080

  7. Surface ultrastructure of the cornea and adjacent epidermis during metamorphosis of Rana pipiens: a scanning electron microscopic study

    SciTech Connect

    Kaltenbach, J.C.; Harding, C.V.; Susan, S.

    1980-01-01

    The external surface of the cornea and adjacent epidermis of larvae in representative developmental stages and of adult frogs, Rana pipiens, was studied by scanning electron microscopy. Surface cells are polygonal, usually hexagonal, in outline and covered with microprojections. During larval development prior to metamorphic stages, neither eyelids nor Harderian glands have developed; microprojections on the corneal surface are high and branched, and cell boundaries are elevated. On the anterior portion of the cornea and on the epidermis near the eye, the surface pattern is less dense, and ciliated cells are present. During metamorphic stages, corneal cell boundaries become less prominent and the pattern of microprojections more variable and markedly different from that of larvae of earlier stages. Corneal cells have a spongy appearance, are covered by a coating material, or are characterized as light or dark based on their brightness and surface texture. As eyelids develop in metamorphic stages XX-XXI, the numbers of ciliated cells increase dramatically, both on the corneal surface and on the edges of the developing lids. In later metamorphic stages XXII to XXV, lids and Harderian glands become well-developed, and cilia are no longer observed. The adjacent epidermal surface becomes devoid of cilia but perforated by openings of cutaneous glands. Its spongy appearance is similar to that of both the cornea and neighboring epidermis of the mature frog. Changes in corneal surface features are probably metamorphic events associated with development of lids and Harderian glands and a shift from an aqueous to an air environment.

  8. Hind limb malformations in free-living northern leopard frogs (Rana pipiens) from Maine, Minnesota, and Vermont suggest multiple etiologies

    USGS Publications Warehouse

    Meteyer, C.U.; Loeffler, I.K.; Fallon, J.F.; Converse, K.A.; Green, E.; Helgen, J.C.; Kersten, S.; Levey, R.; Eaton-Poole, L.; Burkhart, J.G.

    2000-01-01

    Background Reports of malformed frogs have increased throughout the North American continent in recent years. Most of the observed malformations have involved the hind limbs. The goal of this study was to accurately characterize the hind limb malformations in wild frogs as an important step toward understanding the possible etiologies. Methods During 1997 and 1998, 182 recently metamorphosed northern leopard frogs (Rana pipiens) were collected from Minnesota, Vermont, and Maine. Malformed hind limbs were present in 157 (86%) of these frogs, which underwent necropsy and radiographic evaluation at the National Wildlife Health Center. These malformations are described in detail and classified into four major categories: (1) no limb (amelia); (2) multiple limbs or limb elements (polymelia, polydactyly, polyphalangy); (3) reduced limb segments or elements (phocomelia, ectromelia, ectrodactyly, and brachydactyly; and (4) distally complete but malformed limb (bone rotations, bridging, skin webbing, and micromelia). Results Amelia and reduced segments and/or elements were the most common finding. Frogs with bilateral hind limb malformations were not common, and in only eight of these 22 frogs were the malformations symmetrical. Malformations of a given type tended to occur in frogs collected from the same site, but the types of malformations varied widely among all three states, and between study sites within Minnesota. Conclusions Clustering of malformation type suggests that developmental events may produce a variety of phenotypes depending on the timing, sequence, and severity of the environmental insult. Hind limb malformations in free-living frogs transcend current mechanistic explanations of tetrapod limb development.

  9. A physiological measure of shifting connections in the Rana pipiens retinotectal system.

    PubMed

    Fraser, S E; Hunt, R K

    1986-06-01

    The retinotectal connections of developing Rana tadpoles and froglets have been studied using light-pipe techniques to directly assay the pattern of the projection from the retina to the tectum. The projection site of the retina surrounding the optic nerve head was determined at two different stages of development (late larval and metamorphic frog) on the same animal. Small electrolytic marker lesions were used to mark the tectal sites to which the optic nerve head projected at these two times. Comparison of the positions of the two lesions gives a direct measure of the shift in the projection during the interlesion time interval of one week. The results indicate a shift in the projection of 275 micron week-1 in late larval life. Previous work in Xenopus using the light-pipe techniques indicated a qualitatively similar shift during equivalent stages of development, but significantly smaller in magnitude. In the present study, topographic postsynaptic units could be recorded at all stages investigated, indicating functional synapses between the optic nerve fibres and the tectum. Thus, these studies offer evidence of a significant shift in the functional connection pattern of the amphibian retinotectal map during development, in agreement with the recent anatomical data from other laboratories on the Rana and goldfish visual system. PMID:3489803

  10. Octylphenol and UV-B radiation alter larval development and hypothalamic gene expression in the leopard frog (Rana pipiens).

    PubMed Central

    Crump, Douglas; Lean, David; Trudeau, Vance L

    2002-01-01

    We assessed octylphenol (OP), an estrogenic endocrine-disrupting chemical, and UV-B radiation, a known stressor in amphibian development, for their effects on hypothalamic gene expression and premetamorphic development in the leopard frog Rana pipiens. Newly hatched tadpoles were exposed for 10 days to OP alone at two different dose levels; to subambient UV-B radiation alone; and to two combinations of OP and UV-B. Control animals were exposed to ethanol vehicle (0.01%) exposure, a subset of tadpoles from each treatment group was raised to metamorphosis to assess differences in body weight and time required for hindlimb emergence. Tadpoles from one of the OP/UV-B combination groups had greater body weight and earlier hindlimb emergence (p < 0.05), but neither OP nor UV-B alone produced significant changes in body weight or hindlimb emergence, indicating a potential mechanism of interaction between OP and UV-B. We hypothesized that the developing hypothalamus might be a potential environmental sensor for neurotoxicologic studies because of its role in the endocrine control of metamorphosis. We used a differential display strategy to identify candidate genes differentially expressed in the hypothalamic region of the exposed tadpoles. Homology cloning was performed to obtain R. pipiens glutamate decarboxylases--GAD65 and GAD67, enzymes involved in the synthesis of the neurotransmitter gamma-aminobutyric acid (GABA). cDNA expression profiles revealed that OP and UV-B affected the levels of several candidate transcripts in tadpole (i.e., Nck, Ash, and phospholipase C gamma-binding protein 4 and brain angiogenesis inhibitor-3) and metamorph (i.e., GAD67, cytochrome C oxidase, and brain angiogenesis inhibitor-2 and -3) brains. This study represents a novel approach in toxicology that combines physiologic and molecular end points and indicates that levels of OP commonly found in the environment and subambient levels of UV-B alter the expression of important hypothalamic

  11. Alteration of the hypothalamic-pituitary-gonadal axis in estrogen- and androgen-treated adult male leopard frog, Rana pipiens

    PubMed Central

    Tsai, Pei-San; Kessler, Ann E; Jones, Jeremy T; Wahr, Kathleen B

    2005-01-01

    Background Gonadal steroids, in particular 5 alpha-dihydrotestosterone (DHT) and 17 beta-estradiol (E2), have been shown to feed back on the hypothalamic-pituitary-gonadal (HPG) axis of the ranid frog. However, questions still remain on how DHT and E2 impact two of the less-studied components of the ranid HPG axis, the hypothalamus and the gonad, and if the feedback effects are consistently negative. Thus, the goal of the study was to examine the effects of DHT and E2 upon the HPG axis of the gonadally-intact, sexually mature male leopard frogs, Rana pipiens. Methods R. pipiens were implanted with silastic capsules containing either cholesterol (Ch, a control), DHT, or E2 for 10 or 30 days. At each time point, steroid-induced changes in hypothalamic GnRH and pituitary LH concentrations, circulating luteinizing hormone (LH), and testicular histology were examined. Results Frogs implanted with DHT or E2 for 10 days did not show significant alterations in the HPG axis. In contrast, frogs implanted with hormones for 30 days had significantly lower circulating LH (for both DHT and E2), decreased pituitary LH concentration (for E2 only), and disrupted spermatogenesis (for both DHT and E2). The disruption of spermatogenesis was qualitatively similar between DHT and E2, although the effects of E2 were consistently more potent. In both DHT and E2-treated animals, a marked loss of all pre-meiotic germ cells was observed, although the loss of secondary spermatogonia appeared to be the primary cause of disrupted spermatogenesis. Unexpectedly, the presence of post-meiotic germ cells was either unaffected or enhanced by DHT or E2 treatment. Conclusions Overall, these results showed that both DHT and E2 inhibited circulating LH and disrupted spermatogenesis progressively in a time-dependent manner, with the longer duration of treatment producing the more pronounced effects. Further, the feedback effects exerted by both steroid hormones upon the HPG axis were largely negative

  12. Toxicity of the conventional energetics TNT and RDX relative to new insensitive munitions constituents DNAN and NTO in Rana pipiens tadpoles.

    PubMed

    Stanley, Jacob K; Lotufo, Guilherme R; Biedenbach, James M; Chappell, Pornsawan; Gust, Kurt A

    2015-04-01

    An initiative within the US military is targeting the replacement of traditional munitions constituents with insensitive munitions to reduce risk of accidental detonation. The purpose of the present study was to comparatively assess toxicity of the traditional munitions constituents 2,4,6-trinitrotoluene (TNT) and 1,3,5-trinitroperhydro-1,3,5-triazine (RDX) with the new insensitive munitions constituents 2,4-dinitroanisole (DNAN) and 3-nitro-1,2,4-triazol-5-one (NTO). The following exposure durations were performed with Rana pipiens (leopard frog) tadpoles: TNT and DNAN, 96 h and 28 d; RDX, 10 d and 28 d; NTO, 28 d. The 96-h 50% lethal concentration (LC50) values and 95% confidence intervals for TNT and DNAN were 4.4 mg/L (4.2 mg/L, 4. 7 mg/L) and 24.3 mg/L (21.3 mg/L, 27.6 mg/L), respectively. No significant impacts on survival were observed in the 10-d exposure to RDX up to 25.3 mg/L. Effects on tadpole swimming distance were observed with a lowest-observed-effect concentration (LOEC) of 5.9 mg/L RDX. In the 28-d exposures, the LOECs for survival for TNT, DNAN, and NTO were 0.003 mg/L, 2.4 mg/L, and 5.0 mg/L, respectively. No significant mortality was observed in the RDX chronic 28-d exposure up to the highest treatment level tested of 28.0 mg/L. Neither tadpole developmental stage nor growth was significantly affected in any of the 28-d exposures. Rana pipiens were very sensitive to chronic TNT exposure, with an LOEC 3 orders of magnitude lower than those for insensitive munitions constituents DNAN and NTO. PMID:25586961

  13. The disappearing northern leopard frog (Lithobates pipiens): conservation genetics and implications for remnant populations in western Nevada

    PubMed Central

    Rogers, Serena D; Peacock, Mary M

    2012-01-01

    Global amphibian declines suggest a major shift in the amount and quality of habitat for these sensitive taxa. Many species that were once widespread are now experiencing declines either in part of or across their historic range. The northern leopard frog (Rana [Lithobates] pipiens] has undergone significant declines particularly in the western United States and Canada. Leopard frog population losses in Nevada are largely due to habitat fragmentation and the introduction of nonnative fish, amphibian, and plant species. Only two populations remain in the Truckee and Carson River watersheds of western Nevada which represents the western boundary of this species range. We used sequence data for an 812 base pair fragment of the mitochondrial NADH dehydrogenase 1 (ND1) gene to support a native origin for western Nevada populations. All frogs had a single haplotype (W07) from the distinct western North America ND1 haplotype clade. Data from seven polymorphic microsatellite loci show that Truckee and Carson River populations are highly differentiated from each other and from leopard frogs collected from eastern Nevada sites. Lack of gene flow among and distinct color morphs among the western Nevada populations likely predates the current geographical isolation. Comparisons with other peripheral L. pipiens populations show western Nevada populations have similar levels of gene diversity despite their contemporary isolation (HE 0.411, 0.482). Restoration of leopard frog populations in these watersheds will be challenging given well-entrenched nonnative bullfrog populations and major changes to the riparian zone over the past century. Declines of once common amphibian species has become a major conservation concern. Contemporary isolation of populations on a species range periphery such as the leopard frog populations in the Truckee and Carson rivers further exacerbate extirpation risk as these populations are likely to have fewer genetic resources to adaptively respond to

  14. Reproductive interference between Rana dalmatina and Rana temporaria affects reproductive success in natural populations.

    PubMed

    Hettyey, Attila; Vági, Balázs; Kovács, Tibor; Ujszegi, János; Katona, Patrik; Szederkényi, Márk; Pearman, Peter B; Griggio, Matteo; Hoi, Herbert

    2014-10-01

    Experimental evidence suggests that reproductive interference between heterospecifics can seriously affect individual fitness; support from field studies for such an effect has, however, remained scarce. We studied reproductive interference in 25 natural breeding ponds in an area where two ranid frogs, Rana dalmatina and Rana temporaria, co-occur. The breeding seasons of the two species usually overlap and males of both species are often found in amplexus with heterospecific females, even though matings between heterospecifics produce no viable offspring. We estimated species abundance ratios based on the number of clutches laid and evaluated fertilization success. In ponds with low spatial complexity and a species abundance ratio biased towards R. temporaria, the average fertilization success of R. dalmatina eggs decreased, while this relationship was not detectable in spatially more complex ponds. Fertilization success of R. temporaria did not decrease with increasing relative numbers of heterospecifics. This asymmetry in fitness effects of reproductive interference may be attributed to R. temporaria males being more competitive in scramble competition for females than R. dalmatina males. Our study is among the first to demonstrate that in natural breeding populations of vertebrates interference among heterospecifics has the potential to substantially lower reproductive success at the population level, which may in turn affect population dynamics. PMID:25138258

  15. Induction of cytochrome P450-associated monooxygenases in northern leopard frogs, Rana pipiens, by 3,3',4,4',5-pentachlorobiphenyl

    USGS Publications Warehouse

    Huang, Y.-W.; Melancon, M.J.; Jung, R.E.; Karasov, W.H.

    1998-01-01

    Northern leopard frogs (Rana pipiens) were injected intraperitoneally either with a solution of polychlorinated biphenyl (PCB) 126 in corn oil at a concentration of 0.2, 0.7, 2.3 and 7.8 mg/kg body weight or with corn oil alone. Appropriate assay conditions with hepatic microsomes were determined for four cytochrome P450-associated monooxygenases: ethoxyresorufin-O-dealkylase (EROD), methoxy-ROD (MROD), benzyloxy-ROD (BROD) and pentoxy-ROD (PROD). One week after PCB administration, the specific activities of EROD, MROD, BROD and PROD were not elevated at doses ? 0.7 mg/kg (p > 0.05), but were significantly increased at doses ? 2.3 mg/kg compared to the control groups (p < 0.05). The increased activity of these four enzymes ranged from 3to 6.4fold relative to control levels. The increased activities were maintained for at least four weeks. Due to a lack of induction at low doses of PCB 126, which were still relatively high compared to currentlyknown environmental concentrations, we suspect that EROD, MROD, BROD, and PROD activities are not sensitive biomarkers for coplanar PCB exposure in leopard frogs.

  16. Induction of cytochrome P450-associated monooxygenases in northern leopard frogs, Rana pipiens, by 3,3{prime},4,4{prime},5-pentachlorobiphenyl

    SciTech Connect

    Huang, Y.; Jung, R.E.; Karasov, W.H.; Melancon, M.J.

    1998-08-01

    In the past decade, biochemical and physiological characteristics such as hepatic detoxifying system. DNA adducts, thyroid malfunction, and acetylcholinesterase inhibition have been used extensively as biomarkers for contaminant exposure. Northern leopard frogs (Rana pipiens) were injected intraperitoneally either with a solution of polychlorinated biphenyl (PCB) 126 m corn oil at a concentration of 0.2, 0.7, 2.3, or 7.8 mg/kg body weight or with corn oil alone. Appropriate assay conditions with hepatic microsomes were determined for four cytochrome P450-associated monooxygenases: ethoxyresorufin-O-dealkylase (EROD), methoxy-ROD (MROD), benzyloxy-ROD (BROD), and pentoxy-ROD (PROD). One week after PCB administration, the specific activities of EROD, MROD, BROD, and PROD were not elevated at doses {le}0.7 mg/kg (p > 0.05) but were significantly increased at doses {ge}2.3 mg/kg compared to the control groups (p < 0.05). The increased activities of these four enzymes were 3 to 6.4 times those in the control groups. The increased activities were maintained for at least 4 weeks. Because of a lack of induction at low doses of PCB 126, which were still relatively high compared to currently known environmental concentration, the authors suspect that EROD, MROD, BROD, and PROD activities are not sensitive biomarkers for coplanar PCB exposure in leopard frogs.

  17. The effect of fatigue on the binding of glycolytic enzymes in the isolated gastrocnemius of Rana pipiens.

    PubMed

    Guderley, H; Jean, C; Blouin, M

    1989-10-26

    Fatigue of isolated gastrocnemius muscles from R. pipiens leads to a marked increase in the proportion of phosphofructokinase bound to the particulate fraction and a decrease in the binding of lactate dehydrogenase, pyruvate kinase, creatine phosphokinase and glyceraldehyde-3-phosphate dehydrogenase. Only the proportion of aldolase bound to the particulate fraction was unaffected by fatigue. This pattern was unchanged when fatigued muscles were extracted at pH 6.5 rather than 7.5. Thus, muscle fatigue leads to opposite changes in the binding of the glycolytic enzymes. PMID:2804095

  18. Testing of UK Populations of Culex pipiens L. for Schmallenberg Virus Vector Competence and Their Colonization

    PubMed Central

    Manley, Robyn; Harrup, Lara E.; Veronesi, Eva; Stubbins, Francesca; Stoner, Jo; Gubbins, Simon; Wilson, Anthony; Batten, Carrie; Koenraadt, Constantianus J. M.; Henstock, Mark; Barber, James; Carpenter, Simon

    2015-01-01

    Background Schmallenberg virus (SBV), an arboviral pathogen of ruminants, emerged in northern Europe during 2011 and has subsequently spread across a vast geographic area. While Culicoides biting midges (Diptera: Ceratopogonidae) have been identified as a biological transmission agent of SBV, the role of mosquitoes (Diptera: Culicidae) as potential vectors has not been defined beyond small-scale field collections in affected areas. Culex pipiens L. are one of the most widespread mosquitoes in northern Europe; they are present on farms across the region and have previously been implicated as vectors of several other arboviruses. We assessed the ability of three colony lines of Cx. pipiens, originating from geographically diverse field populations, to become fully infected by SBV using semi-quantitative real-time RT-PCR (sqPCR). Findings Two colony lines of Cx. pipiens were created in the UK (‘Brookwood’ and ‘Caldbeck’) from field collections of larvae and pupae and characterised using genetic markers. A third strain of Cx. pipiens from CVI Wageningen, The Netherlands, was also screened during experiments. Intrathoracic inoculation of the Brookwood line resulted in infections after 14 days that were characterised by high levels of RNA throughout individuals, but which demonstrated indirect evidence of salivary gland barriers. Feeding of 322 individuals across the three colony lines on a membrane based infection system resulted in no evidence of full dissemination of SBV, although infections did occur in a small proportion of Cx. pipiens from each line. Conclusions/Significance This study established two novel lines of Cx. pipiens mosquitoes of UK origin in the laboratory and subsequently tested their competence for SBV. Schmallenberg virus replication and dissemination was restricted, demonstrating that Cx. pipiens is unlikely to be an epidemiologically important vector of the virus in northern Europe. PMID:26291533

  19. Wolbachia Endobacteria in Natural Populations of Culex pipiens of Iran and Its Phylogenetic Congruence

    PubMed Central

    Karami, Mohsen; Moosa-Kazemi, Seyed Hassan; Oshaghi, Mohammad Ali; Vatandoost, Hasan; Sedaghat, Mohammad Mehdi; Rajabnia, Ramazan; Hosseini, Mostafa; Maleki-Ravasan, Naseh; Yahyapour, Yousef; Ferdosi-Shahandashti, Elaheh

    2016-01-01

    Background: Wolbachia are common intracellular bacteria that infect different groups of arthropods including mosquitoes. These bacteria modify host biology and may induce feminization, parthenogenesis, male killing and cytoplasmic incompatibility (CI). Recently Wolbachia is being nominated as a bio-agent and paratransgenic candidate to control mosquito borne diseases. Methods: Here we report the results of a survey for presence, frequency, and phylogenetic congruence of these endosymbiont bacteria in Culex pipiens populations in Northern, Central, and Southern parts of Iran using nested-PCR amplification of wsp gene. Results: Wolbachia DNA were found in 227 (87.3%) out of 260 wild-caught mosquitoes. The rate of infection in adult females ranged from 61.5% to 100%, while in males were from 80% to 100%. The Blast search and phylogenetic analysis of the wsp gene sequence revealed that the Wolbachia strain from Iranian Cx. pipiens was identical to the Wolbachia strains of supergroup B previously reported in members of the Cx. pipiens complex. They had also identical sequence homology with the Wolbachia strains from a group of distinct arthropods including lepidopteran, wasps, flies, damselfly, thrips, and mites from remote geographical areas of the world. Conclusion: It is suggested that Wolbachia strains horizontally transfer between unrelated host organisms over evolutionary time. Also results of this study indicates that Wolbachia infections were highly prevalent infecting all Cx. pipiens populations throughout the country, however further study needs to define Wolbachia inter-population reproductive incompatibility pattern and its usefulness as a bio-agent control measure. PMID:27308293

  20. Rapid assay to identify the two genetic forms of Culex (Culex) pipiens L. (Diptera: Culicidae) and hybrid populations.

    PubMed

    Bahnck, Carolyn M; Fonseca, Dina M

    2006-08-01

    A previously developed method to identify members of the Culex pipiens complex exploiting polymorphisms in a nuclear intron (acetylcholinesterase [ACE] based-assay) cannot differentiate the two forms of Cx. pipiens: form pipiens and form molestus. Notably, the two forms seem to differ extensively in behavior and physiology and likely have very different epidemiologic importance. Because they are morphologically indistinguishable, molecular methods are critical for the evaluation of their relative importance. Although the two forms of Cx. pipiens have been distinguished using a panel of microsatellite loci, such a protocol is laborious and expensive. We developed a rapid assay based on polymorphisms in the flanking region of a microsatellite locus. Used in conjunction with the ACE-assay, this new assay allows the identification of pure and hybrid populations of the two Cx. pipiens forms as well as those including Cx. quinquefasciatus. We discuss the usefulness of the method as well as limitations to its application. PMID:16896127

  1. Phenotypic Variation among Culex pipiens Complex (Diptera: Culicidae) Populations from the Sacramento Valley, California: Horizontal and Vertical Transmission of West Nile Virus, Diapause Potential, Autogeny, and Host Selection

    PubMed Central

    Nelms, Brittany M.; Kothera, Linda; Thiemann, Tara; Macedo, Paula A.; Savage, Harry M.; Reisen, William K.

    2013-01-01

    The vector competence and bionomics of Culex pipiens form pipiens L. and Cx. pipiens f. molestus Forskäl were evaluated for populations from the Sacramento Valley. Both f. pipiens and f. molestus females became infected, produced disseminated infections, and were able to transmit West Nile virus. Form molestus females also transmitted West Nile virus vertically to egg rafts and F1 progeny, whereas f. pipiens females only transmitted to egg rafts. Culex pipiens complex from urban Sacramento blood-fed on seven different avian species and two mammalian species. Structure analysis of blood-fed mosquitoes identified K = 4 genetic clusters: f. molestus, f. pipiens, a group of genetically similar hybrids (Cluster X), and admixed individuals. When females were exposed as larvae to midwinter conditions in bioenvironmental chambers, 85% (N = 79) of aboveground Cx. pipiens complex females and 100% (N = 34) of underground f. molestus females did not enter reproductive diapause. PMID:24043690

  2. AMBIENT SOLAR UV RADIATION CAUSES MORTALITY IN LARVAE OF THREE SPECIES OF RANA

    EPA Science Inventory

    Recent reports concerning the lethal effects of solar ultraviolet-B (UV-B) radiation on amphibians suggest that this stressor has the potential to impact some amphibian populations. In this study embryos and larvae of three anuran species, Rana pipiens, R. clamitans, and R. septe...

  3. Hybridization and population structure of the Culex pipiens complex in the islands of Macaronesia

    PubMed Central

    Gomes, Bruno; Alves, Joana; Sousa, Carla A; Santa-Ana, Marta; Vieira, Inês; Silva, Teresa L; Almeida, António PG; Donnelly, Martin J; Pinto, João

    2012-01-01

    The Culex pipiens complex includes two widespread mosquito vector species, Cx. pipiens and Cx. quinquefasciatus. The distribution of these species varies in latitude, with the former being present in temperate regions and the latter in tropical and subtropical regions. However, their distribution range overlaps in certain areas and interspecific hybridization has been documented. Genetic introgression between these species may have epidemiological repercussions for West Nile virus (WNV) transmission. Bayesian clustering analysis based on multilocus genotypes of 12 microsatellites was used to determine levels of hybridization between these two species in Macaronesian islands, the only contact zone described in West Africa. The distribution of the two species reflects both the islands' biogeography and historical aspects of human colonization. Madeira Island displayed a homogenous population of Cx. pipiens, whereas Cape Verde showed a more intriguing scenario with extensive hybridization. In the islands of Brava and Santiago, only Cx. quinquefasciatus was found, while in Fogo and Maio high hybrid rates (∼40%) between the two species were detected. Within the admixed populations, second-generation hybrids (∼50%) were identified suggesting a lack of isolation mechanisms. The observed levels of hybridization may locally potentiate the transmission to humans of zoonotic arboviruses such as WNV. PMID:22957190

  4. Modeling Dynamics of Culex pipiens Complex Populations and Assessing Abatement Strategies for West Nile Virus

    PubMed Central

    Pawelek, Kasia A.; Hager, Elizabeth J.; Hunt, Gregg J.

    2014-01-01

    The primary mosquito species associated with underground stormwater systems in the United States are the Culex pipiens complex species. This group represents important vectors of West Nile virus (WNV) throughout regions of the continental U.S. In this study, we designed a mathematical model and compared it with surveillance data for the Cx. pipiens complex collected in Beaufort County, South Carolina. Based on the best fit of the model to the data, we estimated parameters associated with the effectiveness of public health insecticide (adulticide) treatments (primarily pyrethrin products) as well as the birth, maturation, and death rates of immature and adult Cx. pipiens complex mosquitoes. We used these estimates for modeling the spread of WNV to obtain more reliable disease outbreak predictions and performed numerical simulations to test various mosquito abatement strategies. We demonstrated that insecticide treatments produced significant reductions in the Cx. pipiens complex populations. However, abatement efforts were effective for approximately one day and the vector mosquitoes rebounded until the next treatment. These results suggest that frequent insecticide applications are necessary to control these mosquitoes. We derived the basic reproductive number (ℜ0) to predict the conditions under which disease outbreaks are likely to occur and to evaluate mosquito abatement strategies. We concluded that enhancing the mosquito death rate results in lower values of ℜ0, and if ℜ0<1, then an epidemic will not occur. Our modeling results provide insights about control strategies of the vector populations and, consequently, a potential decrease in the risk of a WNV outbreak. PMID:25268229

  5. PHYLOGEOGRAPHY OF RANA YAVAPAIENSIS AND RANA ONCA: PRELIMINARY FINDINGS WITH CONSERVATION IMPLICATIONS

    EPA Science Inventory

    The closely related aridland frogs Rana onca (Relict Leopard Frog) and Rana yavapaiensis (Lowland Leopard Frog) have both experienced dramatic population declines. Rana onca currently occurs naturally at only 6 disjunct sites in southern Nevada. Rana yavapaiensis is present acros...

  6. POPULATION STATUS AND DISTRIBUTION OF A DECIMATED AMPHIBIAN, THE RELICT LEOPARD FROG (RANA ONCA)

    EPA Science Inventory

    The relict leopard frog (Rana onca) was once thought to be extinct, but has recently been shown to comprise a valid taxon with extant populations. We delineate the minimum historical range of the species, and report results of surveys at 12 historical and 54 other localities to d...

  7. Development of Resistance to Pyrethroid in Culex pipiens pallens Population under Different Insecticide Selection Pressures

    PubMed Central

    Shi, Linna; Hu, Hongxia; Ma, Kai; Zhou, Dan; Yu, Jing; Zhong, Daibin; Fang, Fujin; Chang, Xuelian; Hu, Shengli; Zou, Feifei; Wang, Weijie; Sun, Yan; Shen, Bo; Zhang, Donghui; Ma, Lei; Zhou, Guofa; Yan, Guiyun; Zhu, Changliang

    2015-01-01

    Current vector control programs are largely dependent on pyrethroids, which are the most commonly used and only insecticides recommended by the World Health Organization for insecticide-treated nets (ITNs). However, the rapid spread of pyrethroid resistance worldwide compromises the effectiveness of control programs and threatens public health. Since few new insecticide classes for vector control are anticipated, limiting the development of resistance is crucial for prolonging efficacy of pyrethroids. In this study, we exposed a field-collected population of Culex pipiens pallens to different insecticide selection intensities to dynamically monitor the development of resistance. Moreover, we detected kdr mutations and three detoxification enzyme activities in order to explore the evolutionary mechanism of pyrethroid resistance. Our results revealed that the level of pyrethroid resistance was proportional to the insecticide selection pressure. The kdr and metabolic resistance both contributed to pyrethroid resistance in the Cx. pipiens pallens populations, but they had different roles under different selection pressures. We have provided important evidence for better understanding of the development and mechanisms of pyrethroid resistance which may guide future insecticide use and vector management in order to avoid or delay resistance. PMID:26275298

  8. Seasonal dynamics of insecticide resistance, multiple resistance, and morphometric variation in field populations of Culex pipiens.

    PubMed

    Taskin, Belgin Gocmen; Dogaroglu, Taylan; Kilic, Sercan; Dogac, Ersin; Taskin, Vatan

    2016-05-01

    Resistance to insecticides that impairs nervous transmission has been widely investigated in mosquito populations as insecticides are crucial to effective insect control. The development of insecticide resistance is also of special interest to evolutionary biologists since it represents the opportunity to observe the genetic consequences of a well-characterized alteration in the environment. Although the frequencies of resistance alleles in Culex pipiens populations against different groups of insecticides have been reported, no detailed information is available on the relative change in these allele frequencies over time. In this study, we collected mosquitoes of the Cx. pipiens complex from six locations in three seasons in the Aegean region of Turkey and examined the i) seasonal variations in resistance to four different chemical classes of insecticides, ii) seasonal fluctuations in frequencies of resistance-associated target-site mutations of the three genes (ace-1, kdr, and Rdl), and iii) potential seasonal variations in wing morphometric characters that may be modified in resistant mosquitoes. Our bioassay results indicated the presence of different levels of resistance to all tested insecticides for all three seasons in all locations. The results of the PCR-based molecular analysis revealed low frequencies of mutations in ace-1 and Rdl that are associated with resistance to malathion, bendiocarb, and dieldrin and no obvious seasonal changes. In contrast, we detected high frequencies and striking seasonal changes for two kdr mutations associated with resistance to DDT and pyrethroids. In addition, the evaluation of the field populations from all seasons in terms of the combinations of polymorphisms at four resistance-associated mutations did not reveal the presence of insects that are resistant to all pesticides. Results from the morphological analysis displayed a similar pattern for both wings and did not show a clear separation among the samples from the

  9. Population estimates for the Toiyabe population of the Columbia spotted frog (Rana luteiventris), 2004–10

    USGS Publications Warehouse

    Adams, Michael J.; Mellison, Chad; Galvan, Stephanie K.

    2013-01-01

    The Toiyabe population of Columbia spotted frogs (Rana luteiventris, hereafter "Toiyabe frogs") is a geographically isolated population located in central Nevada (fig. 1). The Toiyabe population is part of the Great Basin Distinct Population Segment of Columbia spotted frogs, and is a candidate for listing under the Endangered Species Act (U.S. Fish and Wildlife Service, 2011). The cluster of breeding sites in central Nevada represents the southernmost extremity of the Columbia spotted frogs' known range (Funk and others, 2008). Toiyabe frogs are known to occur in seven drainages in Nye County, Nevada: Reese River, Cow Canyon Creek, Ledbetter Canyon Creek, Cloverdale Creek, Stewart Creek, Illinois Creek, and Indian Valley Creek. Most of the Toiyabe frog population resides in the Reese River, Indian Valley Creek, and Cloverdale Creek drainages (fig. 1; Nevada Department of Wildlife, 2003). Approximately 90 percent of the Toiyabe frogs' habitat is on public land. Most of the public land habitat (95 percent) is managed by the U.S. Forest Service (USFS), while the Bureau of Land Management (BLM) manages the remainder. Additional Toiyabe frog habitat is under Yomba Shoshone Tribal management and in private ownership (Nevada Department of Wildlife, 2003). The BLM, USFS, Nevada Department of Wildlife (NDOW), Nevada Natural Heritage Program (NNHP), Nye County, and U.S Fish and Wildlife Service (USFWS) have monitored the Toiyabe population since 2004 using mark and recapture surveys (Nevada Department of Wildlife, 2004). The USFWS contracted with the U.S. Geological Survey (USGS) to produce population estimates using these data.

  10. Naturally Occurring Incompatibilities between Different Culex pipiens pallens Populations as the Basis of Potential Mosquito Control Measures

    PubMed Central

    Chen, Lin; Zhu, Changliang; Zhang, Donghui

    2013-01-01

    Background Vector-borne diseases remain a threat to public health, especially in tropical countries. The incompatible insect technique has been explored as a potential control strategy for several important insect vectors. However, this strategy has not been tested in Culex pipiens pallens, the most prevalent mosquito species in China. Previous works used introgression to generate new strains that matched the genetic backgrounds of target populations while harboring a new Wolbachia endosymbiont, resulting in mating competitiveness and cytoplasmic incompatibility. The generation of these incompatible insects is often time-consuming, and the long-term stability of the newly created insect-Wolbachia symbiosis is uncertain. Considering the wide distribution of Cx. pipiens pallens and hence possible isolation of different populations, we sought to test for incompatibilities between natural populations and the possibility of exploiting these incompatibilities as a control strategy. Methodology/Principal Findings Three field populations were collected from three geographic locations in eastern China. Reciprocal cross results showed that bi-directional patterns of incompatibility existed between some populations. Mating competition experiments indicated that incompatible males could compete with cognate males in mating with females, leading to reduced overall fecundity. F1 offspring from incompatible crosses maintained their maternal crossing types. All three populations tested positive for Wolbachia. Removal of Wolbachia by tetracycline rendered matings between these populations fully compatible. Conclusions/Significance Our findings indicate that naturally occurring patterns of cytoplasmic incompatibility between Cx. pipiens pallens populations can be the basis of a control strategy for this important vector species. The observed incompatibilities are caused by Wolbachia. More tests including field trials are warranted to evaluate the feasibility of this strategy as a

  11. Wolbachia and cytoplasmic incompatibility in the California Culex pipiens mosquito species complex: parameter estimates and infection dynamics in natural populations.

    PubMed Central

    Rasgon, Jason L; Scott, Thomas W

    2003-01-01

    Before maternally inherited bacterial symbionts like Wolbachia, which cause cytoplasmic incompatibility (CI; reduced hatch rate) when infected males mate with uninfected females, can be used in a program to control vector-borne diseases it is essential to understand their dynamics of infection in natural arthropod vector populations. Our study had four goals: (1) quantify the number of Wolbachia strains circulating in the California Culex pipiens species complex, (2) investigate Wolbachia infection frequencies and distribution in natural California populations, (3) estimate the parameters that govern Wolbachia spread among Cx. pipiens under laboratory and field conditions, and (4) use these values to estimate equilibrium levels and compare predicted infection prevalence levels to those observed in nature. Strain-specific PCR, wsp gene sequencing, and crossing experiments indicated that a single Wolbachia strain infects Californian Cx. pipiens. Infection frequency was near or at fixation in all populations sampled for 2 years along a >1000-km north-south transect. The combined statewide infection frequency was 99.4%. Incompatible crosses were 100% sterile under laboratory and field conditions. Sterility decreased negligibly with male age in the laboratory. Infection had no significant effect on female fecundity under laboratory or field conditions. Vertical transmission was >99% in the laboratory and approximately 98.6% in the field. Using field data, models predicted that Wolbachia will spread to fixation if infection exceeds an unstable equilibrium point above 1.4%. Our estimates accurately predicted infection frequencies in natural populations. If certain technical hurdles can be overcome, our data indicate that Wolbachia can invade vector populations as part of an applied transgenic strategy for vector-borne disease reduction. PMID:14704183

  12. Population Structure and Distribution Patterns of the Sibling Mosquito Species Culex pipiens and Culex torrentium (Diptera: Culicidae) Reveal Different Evolutionary Paths

    PubMed Central

    Werblow, Antje; Klimpel, Sven; Bolius, Sarah; Dorresteijn, Adriaan W. C.; Sauer, Jan; Melaun, Christian

    2014-01-01

    Nowadays a number of endemic mosquito species are known to possess vector abilities for various diseases, as e.g. the sibling species Culex pipiens and Culex torrentium. Due to their morphological similarity, ecology, distribution and vector abilities, knowledge about these species' population structure is essential. Culicidae from 25 different sampling sites were collected from March till October 2012. All analyses were performed with aligned cox1 sequences with a total length of 658 bp. Population structure as well as distribution patterns of both species were analysed using molecular methods and different statistical tests like distance based redundancy analysis (dbDRA), analysis of molecular variances (AMOVA) or McDonald & Kreitman test and Tajima's D. Within both species, we could show a genetic variability among the cox1 fragment. The construction of haplotype networks revealed one dominating haplotype for Cx. pipiens, widely distributed within Germany and a more homogeneous pattern for Cx. torrentium. The low genetic differences within Cx. pipiens could be a result of an infection with Wolbachia which can induce a sweep through populations by passively taking the also maternally inherited mtDNA through the population, thereby reducing the mitochondrial diversity as an outcome of reproductive incompatibility. Pairwise population genetic differentiation (FST) ranged significantly from moderate to very great between populations of Cx. pipiens and Cx. torrentium. Analyses of molecular variances revealed for both species that the main genetic variability exists within the populations (Cx. pipiens [88.38%]; Cx. torrentium [66.54%]). Based on a distance based redundancy analysis geographical origin explained a small but significant part of the species' genetic variation. Overall, the results confirm that Cx. pipiens and Cx. torrentium underlie different factors regarding their mitochondrial differentiation, which could be a result of endosymbiosis, dispersal

  13. Pond and landscape determinants of Rana dalmatina population sizes in a Romanian rural landscape

    NASA Astrophysics Data System (ADS)

    Hartel, Tibor; Nemes, Szilárd; Cogălniceanu, Dan; Öllerer, Kinga; Moga, Cosmin Ioan; Lesbarrères, David; Demeter, László

    2009-01-01

    Amphibians are good indicators of human impact, declining steadily worldwide. We explored the relationships between the ponds and nearby landscape parameters and population size of the Agile Frog ( Rana dalmatina), estimated from the number of egg masses, in a cultural landscape within the central section of Târnava Mare Basin, Romania. Forty-three permanent ponds were surveyed in a 2600 km 2 area. The average number of egg masses per pond was 211.13 (SD = 426.41). The egg mass number was significantly and positively related to the emergent aquatic macrophyte cover (its effect peaks at around 50%) and the green connecting corridors between the ponds and forests, and negatively related to the extent of nearby urban areas. The proximity of the forest (positive effect) and the presence of high traffic roads (negative effect) were highly correlated with green corridors and further eliminated from the model due to multicollinearity. Both these variables had significant effects when incorporated in univariate models and multivariate models without green corridors. Since a large part of our study area was currently declared as Natura 2000 site, there is an increased need for management proposals and conservation applications for biodiversity, including amphibians. Rana dalmatina is an important species for monitoring because it is common in the studied area and is suited for short surveys.

  14. High Prevalence and Lineage Diversity of Avian Malaria in Wild Populations of Great Tits (Parus major) and Mosquitoes (Culex pipiens)

    PubMed Central

    Glaizot, Olivier; Fumagalli, Luca; Iritano, Katia; Lalubin, Fabrice; Van Rooyen, Juan; Christe, Philippe

    2012-01-01

    Avian malaria studies have taken a prominent place in different aspects of evolutionary ecology. Despite a recent interest in the role of vectors within the complex interaction system of the malaria parasite, they have largely been ignored in most epidemiological studies. Epidemiology of the disease is however strongly related to the vector's ecology and behaviour, and there is a need for basic investigations to obtain a better picture of the natural associations between Plasmodium lineages, vector species and bird hosts. The aim of the present study was to identify the mosquito species involved in the transmission of the haemosporidian parasites Plasmodium spp. in two wild populations of breeding great tits (Parus major) in western Switzerland. Additionally, we compared Plasmodium lineages, based on mitochondrial DNA cytochrome b sequences, between the vertebrate and dipteran hosts, and evaluated the prevalence of the parasite in the mosquito populations. Plasmodium spp. were detected in Culex pipiens only, with an overall 6.6% prevalence. Among the six cytochrome b lineages of Plasmodium identified in the mosquitoes, three were also present in great tits. The results provide evidence for the first time that C. pipiens can act as a natural vector of avian malaria in Europe and yield baseline data for future research on the epidemiology of avian malaria in European countries. PMID:22506060

  15. Irritability Levels of Field and Laboratory Population of Culex pipiens Complex in Tehran to Different Groups of Insecticides

    PubMed Central

    Rahimi, Sara; Vatandoost, Hassan; Abai, Mohammad Reza; Raeisi, Ahmad; Hanafi-Bojd, Ahmad Ali; Rafi, Fatemeh

    2016-01-01

    Background: The irritant effect of some insecticides can cause a proportion of mosquitoes to leave the sprayed rooms before acquiring a lethal dose, so the repeated contact al sub-lethal dose may lead to extent the resistance. Methods: Larvae and pupae of Culex pipiens complex were collected in mass from open canals of waste water in capital city Tehran and reared to obtain the first generation at laboratory. Sugar-fed 2–3 days female mosquitoes were used for the experiments and compared with laboratory strain. The irritability tests of insecticides impregnated papers were measured in plastic conical exposure chambers placed which implemented at controlled conditions according to the method described by WHO. Number of take-offs were counted during 15 minutes of exposure time. Results: DDT had the most irritancy effect against field population of Cx. pipiens. DDT, permethrin and deltamethrin was moderately irritable against laboratory strain, whereas, addition to three previous insecticides, malathion, cyfluthrin and propoxur should be also considered as moderately irritable insecticides for field population of. Irritability level of etofenprox, fenithrothion, bendiocarb, and lambdacyhalothrin did not differ from control group. Conclusion: The irritability response of mosquitoes may have a negative impact on control measures. Periodical execution of irritability tests with insecticides that routinely used in vector control program is highly recommended. PMID:27308276

  16. Population structure of Columbia spotted frogs (Rana luteiventris) is strongly affected by the landscape

    USGS Publications Warehouse

    Funk, W.C.; Blouin, M.S.; Corn, P.S.; Maxell, B.A.; Pilliod, D.S.; Amish, S.; Allendorf, F.W.

    2005-01-01

    Landscape features such as mountains, rivers, and ecological gradients may strongly affect patterns of dispersal and gene flow among populations and thereby shape population dynamics and evolutionary trajectories. The landscape may have a particularly strong effect on patterns of dispersal and gene flow in amphibians because amphibians are thought to have poor dispersal abilities. We examined genetic variation at six microsatellite loci in Columbia spotted frogs (Rana luteiventris) from 28 breeding ponds in western Montana and Idaho, USA, in order to investigate the effects of landscape structure on patterns of gene flow. We were particularly interested in addressing three questions: (i) do ridges act as barriers to gene flow? (ii) is gene flow restricted between low and high elevation ponds? (iii) does a pond equal a 'randomly mating population' (a deme)? We found that mountain ridges and elevational differences were associated with increased genetic differentiation among sites, suggesting that gene flow is restricted by ridges and elevation in this species. We also found that populations of Columbia spotted frogs generally include more than a single pond except for very isolated ponds. There was also evidence for surprisingly high levels of gene flow among low elevation sites separated by large distances. Moreover, genetic variation within populations was strongly negatively correlated with elevation, suggesting effective population sizes are much smaller at high elevation than at low elevation. Our results show that landscape features have a profound effect on patterns of genetic variation in Columbia spotted frogs.

  17. Preexposure to ultraviolet B radiation and 4-tert-octylphenol affects the response of Rana pipiens tadpoles to 3,5,3'-triiodothyronine.

    PubMed

    Croteau, Maxine C; Duarte-Guterman, Paula; Lean, David R S; Trudeau, Vance L

    2010-08-01

    Exposure to multiple environmental stressors is negatively impacting the health of amphibians worldwide. Increased exposure to ultraviolet B radiation (UVBR) and chemical pollutants may affect amphibian populations by disrupting metamorphosis; however, the actual mechanisms by which these stressors affect development remain unknown. Because amphibian metamorphosis is controlled by thyroid hormones (TH), changes in developmental rates by environmental stress suggest a disruption of the thyroid system. Tadpoles were chronically exposed to environmental levels of UVBR (average of 0.15 W/m2) and 4-tert-octylphenol (OP; 10 nM), alone and combined, prior to being challenged to exogenous TH triiodothyronine (T3; 5 or 50 nM). This experimental approach was taken to determine whether exposure to these stressors affects the ability of T3 to elicit specific molecular and morphological responses. Exposure to OP increased mRNA levels of thyroid receptors (TRs) alpha and beta, deiodinase type 2 (D2), and corticotropin releasing hormone in the brain and of D2 in the tail of tadpoles. 4-tert-octylphenol also enhanced T3-induced expression of D2 in the brain. The combination of UVBR and OP affected the expression of TR alpha in the brain and the responses of TR alpha and beta genes to T3 in the tail, demonstrating the importance of considering the effects of multiple stressors on amphibians. Tadpoles exposed to UVBR were developmentally delayed and exhibited slowed tail resorption and accelerated hindlimb development following exposure to T3. Together, these findings indicate that UVBR alters the rate of development and TH-dependent morphological changes at metamorphosis, and that exposure to UVBR and/or OP disrupts the expression of genes important for development and the biological action of T3 in peripheral tissues. Our group is the first to demonstrate that environmental levels of UVBR and/or OP can affect the thyroid system of amphibians. PMID:20821635

  18. Weather factors influencing the population dynamics of Culex pipiens (Diptera: Culicidae) in the Po Plain Valley, Italy (1997-2011).

    PubMed

    Carrieri, Marco; Fariselli, Piero; Maccagnani, Bettina; Angelini, Paola; Calzolari, Mattia; Bellini, Romeo

    2014-04-01

    The impact of weather variables on Culex pipiens L. (Diptera: Culicidae) population dynamics in the Po Valley, Northern Italy, a densely populated region containing the largest industrial and agricultural areas in Italy, was investigated. Monitoring of mosquitoes was carried out by using CO(2)-baited traps without light, collecting data weekly from 1700 to 0900 hours during the period May-September, from 1997 to 2011. Daily minimum, average, and maximum relative humidity; daily minimum, maximum, and average temperature; rainfall; and hydroclimatic balance (rainfall-potential evapotranspiration) were obtained from three weather stations within the surveillance zone. The average population dynamic trend over the 15-yr period showed a bell-shaped curve with a major peak in June and a secondary peak at the end of August in the rural areas, whereas bimodality was not evidenced in the urban areas. The correlation analyses showed that the mosquito seasonal population and the population in the period of maximum West Nile virus circulation (August-September) was mostly affected by the relative humidity registered from March to July, particularly in May, and, to a lower extent, also by hydroclimatic balance registered in April-July, and by the rainfall occurred in June-July. In addition, the rate of increase of the population during the spring months influenced the development of the mosquito population of the following months. PMID:24763101

  19. Determinants of the population growth of the West Nile virus mosquito vector Culex pipiens in a repeatedly affected area in Italy

    PubMed Central

    2014-01-01

    Background The recent spread of West Nile Virus in temperate countries has raised concern. Predicting the likelihood of transmission is crucial to ascertain the threat to Public and Veterinary Health. However, accurate models of West Nile Virus (WNV) expansion in Europe may be hampered by limited understanding of the population dynamics of their primary mosquito vectors and their response to environmental changes. Methods We used data collected in north-eastern Italy (2009–2011) to analyze the determinants of the population growth rate of the primary WNV vector Culex pipiens. A series of alternative growth models were fitted to longitudinal data on mosquito abundance to evaluate the strength of evidence for regulation by intrinsic density-dependent and/or extrinsic environmental factors. Model-averaging algorithms were then used to estimate the relative importance of intrinsic and extrinsic variables in describing the variations of per-capita growth rates. Results Results indicate a much greater contribution of density-dependence in regulating vector population growth rates than of any environmental factor on its own. Analysis of an average model of Cx. pipiens growth revealed that the most significant predictors of their population dynamics was the length of daylight, estimated population size and temperature conditions in the 15 day period prior to sampling. Other extrinsic variables (including measures of precipitation, number of rainy days, and humidity) had only a minor influence on Cx. pipiens growth rates. Conclusions These results indicate the need to incorporate density dependence in combination with key environmental factors for robust prediction of Cx. pipiens population expansion and WNV transmission risk. We hypothesize that detailed analysis of the determinants of mosquito vector growth rate as conducted here can help identify when and where an increase in vector population size and associated WNV transmission risk should be expected. PMID:24428887

  20. Transposon-mediated resistance to Bacillus sphaericus in a field-evolved population of Culex pipiens (Diptera: Culicidae).

    PubMed

    Darboux, Isabelle; Charles, Jean-François; Pauchet, Yannick; Warot, Sylvie; Pauron, David

    2007-08-01

    The binary toxin is the major active component of Bacillus sphaericus, a microbial larvicide used for controlling some vector mosquito-borne diseases. B. sphaericus resistance has been reported in many part of the world, leading to a growing concern for the usefulness of this environmental friendly insecticide. Here we characterize a novel mechanism of resistance to the binary toxin in a natural population of the West Nile virus vector, Culex pipiens. We show that the insertion of a transposable element-like DNA into the coding sequence of the midgut toxin receptor induces a new mRNA splicing event, unmasking cryptic donor and acceptor sites located in the host gene. The creation of the new intron causes the expression of an altered membrane protein, which is incapable of interacting with the toxin, thus providing the host mosquito with an advantageous phenotype. As a large portion of insect genomes is composed of transposable elements or transposable elements-related sequences, this new mechanism may be of general importance to appreciate their significance as potent agents for insect resistance to the microbial insecticides. PMID:17394558

  1. STATUS OF THE RELICT LEOPARD FROG (RANA ONCA): OUR LIMITED UNDERSTANDING OF THE DISTRIBUTION, SIZE, AND DYNAMICS OF EXTANT AND RECENTLY EXTINCT POPULATIONS

    EPA Science Inventory

    The relict leopard frog (Rana onca) was once thought to be extinct, but has recently been shown to comprise a valid taxon with extant populations. Here, we discuss research from several studies, conducted between 1991 and 200 1, that represent the basis for our understanding of t...

  2. Removal of nonnative fish results in population expansion of a declining amphibian (mountain yellow-legged frog, Rana muscosa).

    PubMed

    Knapp, Roland A; Boiano, Daniel M; Vredenburg, Vance T

    2007-02-01

    The mountain yellow-legged frog (Rana muscosa) was once a common inhabitant of the Sierra Nevada (California, USA), but has declined precipitously during the past century due in part to the introduction of nonnative fish into naturally fishless habitats. The objectives of the current study were to describe (1) the effect of fish removal from three lakes (located in two watersheds) on the small, remnant R. muscosa populations inhabiting those lakes, and (2) the initial development of metapopulation structure in each watershed as R. muscosa from expanding populations in fish-removal lakes dispersed to adjacent habitats. At all three fish-removal lakes, R. muscosa population densities increased significantly following the removal of predatory fish. The magnitude of these increases was significantly greater than that observed over the same time period in R. muscosa populations inhabiting control lakes that remained in their natural fishless condition. Following these population increases, R. muscosa dispersed to adjacent suitable (but unoccupied) sites, moving between 200 and 900 m along streams or across dry land. Together, these results suggest that large-scale removal of introduced fish could result in at least partial reversal of the decline of R. muscosa. Continued monitoring of R. muscosa at the fish-removal sites will be necessary to determine whether the positive effects of fish eradication are sustained over the long-term, especially in light of the increasingly important role played by an emerging infectious disease (chytridiomycosis, caused by Batrachochytrium dendrobatidis) in influencing R. muscosa populations. PMID:17396156

  3. Genetic variation in insecticide tolerance in a population of southern leopard frogs (Rana sphenocephala): Implications for amphibian conservation

    USGS Publications Warehouse

    Bridges, C.M.; Semlitsch, R.D.

    2001-01-01

    Currently, conservation efforts are devoted to determining the extent and the causes of the decline of many amphibian species worldwide. Human impacts frequently degrade amphibian habitat and have been implicated in many declines. Because genetic variance is critical in determining the persistence of a species in a changing environment, we examined the amount of genetic variability present in a single population for tolerance to an environmental stressor. We examined the amount of genetic variability among full- and half-sib families in a single population of southern leopard frogs (Rana sphenocephala) with respect to their tolerance to lethal concentrations of the agricultural chemical, carbaryl. Analysis of time-to-death data indicated significant differences among full-sib families and suggests a large amount of variability present in the responses to this environmental stressor. Significant differences in responses among half-sib families indicated that there is additive genetic variance. These data suggest that this population may have the ability to adapt to environmental stressors. It is possible that declines of amphibian populations in the western United States may be attributed to low genetic variability resulting from limited migration among populations and small population sizes.

  4. The Role of Climatic and Density Dependent Factors in Shaping Mosquito Population Dynamics: The Case of Culex pipiens in Northwestern Italy

    PubMed Central

    Giacobini, Mario; Pugliese, Andrea; Merler, Stefano; Rosà, Roberto

    2016-01-01

    Culex pipiens mosquito is a species widely spread across Europe and represents a competent vector for many arboviruses such as West Nile virus (WNV), which has been recently circulating in many European countries, causing hundreds of human cases. In order to identify the main determinants of the high heterogeneity in Cx. pipiens abundance observed in Piedmont region (Northwestern Italy) among different seasons, we developed a density-dependent stochastic model that takes explicitly into account the role played by temperature, which affects both developmental and mortality rates of different life stages. The model was calibrated with a Markov chain Monte Carlo approach exploring the likelihood of recorded capture data gathered in the study area from 2000 to 2011; in this way, we disentangled the role played by different seasonal eco-climatic factors in shaping the vector abundance. Illustrative simulations have been performed to forecast likely changes if temperature or density–dependent inputs would change. Our analysis suggests that inter-seasonal differences in the mosquito dynamics are largely driven by different temporal patterns of temperature and seasonal-specific larval carrying capacities. Specifically, high temperatures during early spring hasten the onset of the breeding season and increase population abundance in that period, while, high temperatures during the summer can decrease population size by increasing adult mortality. Higher densities of adult mosquitoes are associated with higher larval carrying capacities, which are positively correlated with spring precipitations. Finally, an increase in larval carrying capacity is expected to proportionally increase adult mosquito abundance. PMID:27105065

  5. The Role of Climatic and Density Dependent Factors in Shaping Mosquito Population Dynamics: The Case of Culex pipiens in Northwestern Italy.

    PubMed

    Marini, Giovanni; Poletti, Piero; Giacobini, Mario; Pugliese, Andrea; Merler, Stefano; Rosà, Roberto

    2016-01-01

    Culex pipiens mosquito is a species widely spread across Europe and represents a competent vector for many arboviruses such as West Nile virus (WNV), which has been recently circulating in many European countries, causing hundreds of human cases. In order to identify the main determinants of the high heterogeneity in Cx. pipiens abundance observed in Piedmont region (Northwestern Italy) among different seasons, we developed a density-dependent stochastic model that takes explicitly into account the role played by temperature, which affects both developmental and mortality rates of different life stages. The model was calibrated with a Markov chain Monte Carlo approach exploring the likelihood of recorded capture data gathered in the study area from 2000 to 2011; in this way, we disentangled the role played by different seasonal eco-climatic factors in shaping the vector abundance. Illustrative simulations have been performed to forecast likely changes if temperature or density-dependent inputs would change. Our analysis suggests that inter-seasonal differences in the mosquito dynamics are largely driven by different temporal patterns of temperature and seasonal-specific larval carrying capacities. Specifically, high temperatures during early spring hasten the onset of the breeding season and increase population abundance in that period, while, high temperatures during the summer can decrease population size by increasing adult mortality. Higher densities of adult mosquitoes are associated with higher larval carrying capacities, which are positively correlated with spring precipitations. Finally, an increase in larval carrying capacity is expected to proportionally increase adult mosquito abundance. PMID:27105065

  6. Culex pipiens and Stegomyia albopicta (= Aedes albopictus) populations as vectors for lineage 1 and 2 West Nile virus in Europe.

    PubMed

    Brustolin, M; Talavera, S; Santamaría, C; Rivas, R; Pujol, N; Aranda, C; Marquès, E; Valle, M; Verdún, M; Pagès, N; Busquets, N

    2016-06-01

    The emerging disease West Nile fever is caused by West Nile virus (WNV), one of the most widespread arboviruses. This study represents the first test of the vectorial competence of European Culex pipiens Linnaeus 1758 and Stegomyia albopicta (= Aedes albopictus) (both: Diptera: Culicidae) populations for lineage 1 and 2 WNV isolated in Europe. Culex pipiens and S. albopicta populations were susceptible to WNV infection, had disseminated infection, and were capable of transmitting both WNV lineages. This is the first WNV competence assay to maintain mosquito specimens under environmental conditions mimicking the field (day/night) conditions associated with the period of maximum expected WNV activity. The importance of environmental conditions is discussed and the issue of how previous experiments conducted in fixed high temperatures may have overestimated WNV vector competence results with respect to natural environmental conditions is analysed. The information presented should be useful to policymakers and public health authorities for establishing effective WNV surveillance and vector control programmes. This would improve preparedness to prevent future outbreaks. PMID:26890285

  7. Demography and movement in a relocated population of Oregon Spotted Frogs (Rana pretiosa): Influence of season and gender

    USGS Publications Warehouse

    Chelgren, N.D.; Pearl, C.A.; Adams, M.J.; Bowerman, J.

    2008-01-01

    We used five years of recapture data and Bayesian estimation to assess seasonal survival, movement, and growth of Oregon Spotted Frogs (Rana pretiosa) relocated into created ponds at Dilman Meadow in Oregon, USA. We evaluate hypotheses specific to the relocation and elucidate aspects of R. pretiosa life history that are poorly known. The odds of survival of relocated individuals during the first year following relocation were 0.36 times the survival odds of relocated and non-relocated frogs after one year since the relocation. Survival rate was higher for large frogs. After accounting for frog size, we found little variation in survival between ponds at Dilman Meadow. Survival was lowest for males during the breeding/post-breeding redistribution period, suggesting a high cost of breeding for males. The highest survival rates occurred during winter for both genders, and one small spring was used heavily during winter but was used rarely during the rest of the year. Individual growth was higher in ponds that were not used for breeding, and increased with increasing pond age. Our study supports other evidence that R. pretiosa use different habitats seasonally and are specific in their overwintering habitat requirements. Because frogs were concentrated during winter, predator-free overwintering springs are likely to be of particular value for R. pretiosa populations. ?? 2008 by the American Society of Ichthyologists and Herpetologists.

  8. Asymmetric introgression between sympatric molestus and pipiens forms of Culex pipiens (Diptera: Culicidae) in the Comporta region, Portugal

    PubMed Central

    2009-01-01

    Background Culex pipiens L. is the most widespread mosquito vector in temperate regions. This species consists of two forms, denoted molestus and pipiens, that exhibit important behavioural and physiological differences. The evolutionary relationships and taxonomic status of these forms remain unclear. In northern European latitudes molestus and pipiens populations occupy different habitats (underground vs. aboveground), a separation that most likely promotes genetic isolation between forms. However, the same does not hold in southern Europe where both forms occur aboveground in sympatry. In these southern habitats, the extent of hybridisation and its impact on the extent of genetic divergence between forms under sympatric conditions has not been clarified. For this purpose, we have used phenotypic and genetic data to characterise Cx. pipiens collected aboveground in Portugal. Our aims were to determine levels of genetic differentiation and the degree of hybridisation between forms occurring in sympatry, and to relate these with both evolutionary and epidemiological tenets of this biological group. Results Autogeny and stenogamy was evaluated in the F1 progeny of 145 individual Cx. pipiens females. Bayesian clustering analysis based on the genotypes of 13 microsatellites revealed two distinct genetic clusters that were highly correlated with the alternative traits that define pipiens and molestus. Admixture analysis yielded hybrid rate estimates of 8-10%. Higher proportions of admixture were observed in pipiens individuals suggesting that more molestus genes are being introgressed into the pipiens form than the opposite. Conclusion Both physiological/behavioural and genetic data provide evidence for the sympatric occurrence of molestus and pipiens forms of Cx. pipiens in the study area. In spite of the significant genetic differentiation between forms, hybridisation occurs at considerable levels. The observed pattern of asymmetric introgression probably relates to

  9. Extinction of montane populations of the northern leopard frog (Rana pippins) in Colorado

    USGS Publications Warehouse

    Corn, Paul Stephen; Fogleman, James C.

    1984-01-01

    Between 1973 and 1982 nine populations of the northern leopard frog in the Red Feather Lakes region of Larimer County, Colorado, failed in reproduce. These failures all resulted in extinction of the populations. One area formerly supporting a population was recolonized in 1980, but no frogs were observed at any of the nine sites in 1981 or 1982. Six of the populations went extinct because the breeding ponds dried up. The remaining populations were small enough to be susceptible to random events, but the nature of these events is unknown.

  10. PHOTOINDUCED TOXICITY OF FLUORANTHENE TO LARVAE OF THE LEOPARD FROG (RANA PIPENS)

    EPA Science Inventory

    Rana pipiens larvae (96-118 hr old) were exposed to in a flow-through diluter system to five concentrations of fluoranthene for 48 hr. Following the uptake period the exposed larvae were divided into three groups: one for tissue residue analysis, a second for residue analysis fo...

  11. Diversity of Wolbachia pipientis Strain wPip in a Genetically Admixtured, Above-Ground Culex pipiens (Diptera: Culicidae) Population: Association With Form Molestus Ancestry and Host Selection Patterns

    PubMed Central

    MORNINGSTAR, REBECCA J.; HAMER, GABRIEL L.; GOLDBERG, TONY L.; HUANG, SHAOMING; ANDREADIS, THEODORE G.; WALKER, EDWARD D.

    2014-01-01

    Analysis of molecular genetic diversity in nine marker regions of five genes within the bacteriophage WO genomic region revealed high diversity of the Wolbachia pipentis strain wPip in a population of Culex pipiens L. sampled in metropolitan Chicago, IL. From 166 blood fed females, 50 distinct genetic profiles of wPip were identified. Rarefaction analysis suggested a maximum of 110 profiles out of a possible 512 predicted by combinations of the nine markers. A rank-abundance curve showed that few strains were common and most were rare. Multiple regression showed that markers associated with gene Gp2d, encoding a partial putative capsid protein, were significantly associated with ancestry of individuals either to form molestus or form pipiens, as determined by prior microsatellite allele frequency analysis. None of the other eight markers was associated with ancestry to either form, nor to ancestry to Cx. quinquefasciatus Say. Logistic regression of host choice (mammal vs. avian) as determined by bloodmeal analysis revealed that significantly fewer individuals that had fed on mammals had the Gp9a genetic marker (58.5%) compared with avian-fed individuals (88.1%). These data suggest that certain wPip molecular genetic types are associated with genetic admixturing in the Cx. pipiens complex of metropolitan Chicago, IL, and that the association extends to phenotypic variation related to host preference. PMID:22679853

  12. ELECTRON MICROSCOPY OF GROWING OOCYTES OF RANA PIPIENS

    PubMed Central

    Kemp, Norman E.

    1956-01-01

    1. In the cytoplasm of oocytes of stage Y0, prior to the appearance of yolk, one observes a few scattered profiles of endoplasmic reticulum and numerous filamentous mitochondria, usually distributed at random but sometimes clustered. As the nuclear membrane begins to bulge outward, small granules and short rods appear in the perinuclear cytoplasm and endoplasmic reticulum becomes more prominent throughout the cytoplasm. 2. Coincident with the appearance of the first yolk platelets, which are deposited in a narrow peripheral ring within the endoplasm at stage Y1, protoplasmic processes, the microvilli, push out all over the surface of the oocyte. At the same time follicle cells pull away but remain attached to the oocyte at some points through finger-like processes which interdigitate with neighboring microvilli. It is estimated that the microvilli increase the absorptive area of the surface to about thirty-five times that of a simple sphere. Just beneath the microvillous layer is the basal protoplasm of the cortex, now containing tiny granules probably synthesized from newly absorbed raw materials. Cortical granules appear and become aligned below the basal layer on the external border of the endoplasm. Both the cortical granules and the yolk platelets measure up to 1 µ in diameter at this stage. 3. By stage Y3 (yolk filling peripheral three-fourths of cytoplasm), the basal layer of the cortex is folded so that it appears in section as alternating ridges and valleys. The microvilli now extend from the summits of the cortical ridges. Small, ring-shaped granules are abundant in the cortex. Cortical granules have increased to 2 µ in diameter. 4. Yolk platelets continue to be synthesized around the cortical granules and in the subjacent endoplasm. The largest platelets measured in the interior cytoplasm at stage Y4 (cytoplasm filled with yolk) were 3.7 µ wide by 5.8 µ long. Pigment granules increase in size from 0.15 µ in diameter at stage Y3 to 0.30 µ in diameter at stage Y4. PMID:13331960

  13. Evolutionary history of the northern leopard frog: reconstruction of phylogeny, phylogeography, and historical changes in population demography from mitochondrial DNA.

    PubMed

    Hoffman, Eric A; Blouin, Michael S

    2004-01-01

    This study uses a combined methodological approach including phylogenetic, phylogeographic, and demographic analyses to understand the evolutionary history of the northern leopard frog, Rana pipiens. We tested hypotheses concerning how (or if) known geological events and key features of the species biology influenced the contemporary geographic and genetic distribution of R. pipiens. We assayed mitochondrial DNA variation from 389 individuals within 35 populations located throughout the species range. Our a priori expectations for patterns and processes influencing the current genetic structure of R. pipiens were supported by the data. However, our analyses revealed specific aspects of R. pipiens evolutionary history that were unexpected. The phylogenetic analysis indicated that R. pipiens is split into populations containing discrete eastern or western haplotypes, with the Mississippi River and Great Lakes region dividing the geographic ranges. Nested clade analysis indicated that the biological process most often invoked to explain the pattern of haplotype position is restricted gene flow with isolation by distance. Demographic analyses showed evidence of both historical bottlenecks and population expansions. Surprisingly, the genetic evidence indicated that the western haplotypes had significantly reduced levels of genetic diversity relative to the eastern haplotypes and that major range expansions occurred in both regions well before the most recent glacial retreat. This study provides a detailed history of how a widespread terrestrial vertebrate responded to episodic Pleistocene glacial events in North America. Moreover, this study illustrates how complementary methods of data analysis can be used to disentangle recent and ancient effects on the genetic structure of a species. PMID:15058727

  14. Experimental investigation of the susceptibility of Italian Culex pipiens mosquitoes to Zika virus infection.

    PubMed

    Boccolini, Daniela; Toma, Luciano; Di Luca, Marco; Severini, Francesco; Romi, R; Remoli, Maria Elena; Sabbatucci, Michela; Venturi, Giulietta; Rezza, Giovanni; Fortuna, Claudia

    2016-09-01

    We investigated the susceptibility of an Italian population of Culex pipiens mosquitoes to Zika virus (ZIKV) infection, tested in parallel with Aedes aegypti, as a positive control. We analysed mosquitoes at 0, 3, 7, 10, 14, 20 and 24 days after an infectious blood meal. Viral RNA was detected in the body of Cx. pipiens up to three days post-infection, but not at later time points. Our results indicate that Cx. pipiens is not susceptible to ZIKV infection. PMID:27605056

  15. Response of the Italian agile frog (Rana latastei) to a Ranavirus, frog virus 3: a model for viral emergence in naïve populations.

    PubMed

    Pearman, Peter B; Garner, Trenton W J; Straub, Monika; Greber, Urs F

    2004-10-01

    Ranavirus (family Iridoviridae) is a genus of pathogens of poikilotherms, and some ranaviruses may play a role in widespread mortality of amphibians. Ecology of viral transmission in amphibians is poorly known but can be addressed through experimentation in the laboratory. In this study, we use the Ranavirus frog virus 3 (FV3) as an experimental model for pathogen emergence in naive populations of tadpoles. We simulated emerging disease by exposing tadpoles of the Italian agile frog (Rana latastei), to the North American Ranavirus FV3. We demonstrated that mortality occurred due to viral exposure, exposure of tadpoles to decreasing concentrations of FV3 in the laboratory produced dose-dependent survival rates, and cannibalism of virus-carrying carcasses increased mortality due to FV3. These experiments suggest the potential for ecological mechanisms to affect the level of exposure of tadpoles to Ranavirus and to impact transmission of viral pathogens in aquatic systems. PMID:15650083

  16. Feeding patterns of molestus and pipiens forms of Culex pipiens (Diptera: Culicidae) in a region of high hybridization

    PubMed Central

    2013-01-01

    Background Two biological forms of the mosquito Culex pipiens s.s., denoted pipiens and molestus, display behavioural differences that may affect their role as vectors of arboviruses. In this study, the feeding patterns of molestus and pipiens forms were investigated in Comporta (Portugal), where high levels of inter-form admixture have been recorded. Methods Indoor and outdoor mosquito collections were performed in the summer of 2010. Collected Cx. pipiens s.l. females were molecularly identified to species and form by PCR and genotyped for six microsatellites. The source of the blood meal in post-fed females was determined by ELISA and mitochondrial DNA sequencing. Results The distribution of the forms differed according to the collection method. The molestus form was present only in indoor collections, whereas pipiens and admixed individuals were sampled both indoors and outdoors. In both forms, over 90% of blood meals were made on avian hosts. These included blood meals taken from Passeriformes (Passer domesticus and Turdus merula) by females caught resting inside domestic shelters. Conclusion Genetic structure and blood meal analyses suggest the presence of a bird biting molestus population in the study area. Both forms were found to rest indoors, mainly in avian shelters, but at least a proportion of females of the pipiens form may bite outdoors in sylvan habitats and then search for anthropogenic resting sites to complete their gonotrophic cycle. This behaviour may potentiate the accidental transmission of arboviruses to humans in the region. PMID:23578139

  17. Population declines lead to replicate patterns of internal range structure at the tips of the distribution of the California red-legged frog (Rana draytonii)

    USGS Publications Warehouse

    Richmond, Jonathan Q.; Backlin, Adam R.; Tatarian, Patricia J.; Solvesky, Ben G.; Fisher, Robert N.

    2014-01-01

    Demographic declines and increased isolation of peripheral populations of the threatened California red-legged frog (Rana draytonii) have led to the formation of internal range boundaries at opposite ends of the species’ distribution. While the population genetics of the southern internal boundary has been studied in some detail, similar information is lacking for the northern part of the range. In this study, we used microsatellite and mtDNA data to examine the genetic structuring and diversity of some of the last remaining R. draytonii populations in the northern Sierra Nevada, which collectively form the northern external range boundary. We compared these data to coastal populations in the San Francisco Bay Area, where the species is notably more abundant and still exists throughout much of its historic range. We show that ‘external’ Sierra Nevada populations have lower genetic diversity and are more differentiated from one another than their ‘internal’ Bay Area counterparts. This same pattern was mirrored across the distribution in California, where Sierra Nevada and Bay Area populations had lower allelic variability compared to those previously studied in coastal southern California. This genetic signature of northward range expansion was mirrored in the phylogeography of mtDNA haplotypes; northern Sierra Nevada haplotypes showed greater similarity to haplotypes from the south Coast Ranges than to the more geographically proximate populations in the Bay Area. These data cast new light on the geographic origins of Sierra Nevada R. draytonii populations and highlight the importance of distinguishing the genetic effects of contemporary demographic declines from underlying signatures of historic range expansion when addressing the most immediate threats to population persistence. Because there is no evidence of contemporary gene flow between any of the Sierra Nevada R. draytonii populations, we suggest that management activities should focus on

  18. EVIDENCE OF PHYLOGENETICALLY DISTINCT LEOPARD FROGS (RANA ONCA) FROM THE BORDER REGION OF NEVADA, UTAH, AND ARIZONA

    EPA Science Inventory

    Remnant populations of leopard frogs within the Virgin River drainage and adjacent portions of the Colorado River (Black Canyon) in northwestern Arizona and southern Nevada either represent the reportedly extinct taxon Rana onca or northern, disjunct Rana yavapaiensis. To determi...

  19. Daily blood feeding rhythms of laboratory-reared North American Culex pipiens

    PubMed Central

    2014-01-01

    Background Blood feeding by free-living insect vectors of disease is rhythmic and can be used to predict when infectious bites will occur. These daily rhythms can also be targeted by control measures, as in insecticide-treated nets. Culex pipiens form pipiens and C.p. f. molestus are two members of the Culex pipiens assemblage and vectors of West Nile Virus throughout North America. Although Culex species vector human pathogens and parasites, the daily blood feeding rhythms of C.p. f. molestus, to our knowledge, have not been studied. We described and compared the daily blood feeding rhythms of three laboratory-reared populations of Culex pipiens, one of which has confirmed molestus ancestry. We also examined the plasticity of blood feeding time for these three populations. Results For most (>70%) C.p. f. pipiens and C.p. f. molestus collected from metropolitan Chicago, IL, blood feeding took place during scotophase. Peak blood feeding occurred in mid-scotophase, 3-6 hours after lights off. For C.p. f. pipiens originating from Pennsylvania, most mosquitoes (> 90%) blood fed during late photophase and early scotophase. C.p. f. molestus denied a blood meal during scotophase were less likely to blood feed during early photophase (< 20%) than were C.p. f. pipiens from Chicago (> 50%). C.p. f. pipiens from Pennsylvania were capable of feeding readily at any hour of photo- or scotophase. Conclusions Daily blood feeding rhythms of C.p. f. molestus are similar to those of C.p. f. pipiens, particularly when populations originate from the same geographic region. However, the timing of blood feeding is more flexible for C.p. f. pipiens populations relative to C.p. f. molestus. PMID:24450879

  20. Pseudacris triseriata (western chorus frog) and Rana sylvatica (wood frog) chytridiomycosis

    USGS Publications Warehouse

    Rittman, S.E.; Muths, E.; Green, D.E.

    2003-01-01

    The chytrid fungus Batrachochytrium dendrobatidis is a known pathogen of anuran amphibians, and has been correlated with amphibian die-offs worldwide (Daszak et. al. 1999. Emerging Infectious Diseases 5:735-748). In Colorado, B. dendrobatidis has infected Boreal toads (Bufo boreas) (Muths et. al., in review) and has been identified on museum specimens of northern leopard frogs (Rana pipiens) (Carey et. al. 1999. Develop. Comp. Immunol. 23:459-472). We report the first verified case of chytrid fungus in chorus frogs (Pseudacris triseriata) and wood frogs (Rana sylvatica) in the United States. We collected seven P. triseriata, and two adult and two juvenile R. sylvatica in the Kawuneeche Valley in Rocky Mountain National Park (RMNP) during June 2001. These animals were submitted to the National Wildlife Health Center (NWHC) as part of an amphibian health evaluation in RMNP. Chorus frogs were shipped in one container. Wood frog adults and juveniles were shipped in two separate containers. Histological examinations of all chorus frogs and 3 of 4 wood frogs were positive for chytrid fungus infection. The fourth (adult) wood frog was too decomposed for meaningful histology. Histological findings consisted of multifocally mild to diffusely severe infections of the epidermis of the ventrum and hindlimb digital skin. Chytrid thalli were confined to the thickened epidermis (hyperkeratosis), were spherical to oval, and occasional thalli contained characteristic discharge pores or zoospores (Green and Kagarise Sherman 1999. J. Herpetol 35:92-103; Fellers et al. 2001. Copeia 2001:945-953). We cannot confirm that all specimens carried the fungus at collection, because infection may have spread from one individual to all other individuals in each container during transport. Further sampling of amphibians in Kawuneeche Valley is warranted to determine the rate of infection and mortality in these populations.

  1. The genetic contribution to sex determination and number of sex chromosomes vary among populations of common frogs (Rana temporaria).

    PubMed

    Rodrigues, N; Vuille, Y; Brelsford, A; Merilä, J; Perrin, N

    2016-07-01

    The patterns of sex determination and sex differentiation have been shown to differ among geographic populations of common frogs. Notably, the association between phenotypic sex and linkage group 2 (LG2) has been found to be perfect in a northern Swedish population, but weak and variable among families in a southern one. By analyzing these populations with markers from other linkage groups, we bring two new insights: (1) the variance in phenotypic sex not accounted for by LG2 in the southern population could not be assigned to genetic factors on other linkage groups, suggesting an epigenetic component to sex determination; (2) a second linkage group (LG7) was found to co-segregate with sex and LG2 in the northern population. Given the very short timeframe since post-glacial colonization (in the order of 1000 generations) and its seemingly localized distribution, this neo-sex chromosome system might be the youngest one described so far. It does not result from a fusion, but more likely from a reciprocal translocation between the original Y chromosome (LG2) and an autosome (LG7), causing their co-segregation during male meiosis. By generating a strict linkage between several important genes from the sex-determination cascade (Dmrt1, Amh and Amhr2), this neo-sex chromosome possibly contributes to the 'differentiated sex race' syndrome (strictly genetic sex determination and early gonadal development) that characterizes this northern population. PMID:27071845

  2. Ecological Distribution and CQ11 Genetic Structure of Culex pipiens Complex (Diptera: Culicidae) in Italy

    PubMed Central

    Di Luca, Marco; Toma, Luciano; Boccolini, Daniela; Severini, Francesco; La Rosa, Giuseppe; Minelli, Giada; Bongiorno, Gioia; Montarsi, Fabrizio; Arnoldi, Daniele; Capelli, Gioia; Rizzoli, Annapaola; Romi, Roberto

    2016-01-01

    Mosquitoes in the Culex pipiens complex are considered to be involved in the transmission of a range of pathogens, including West Nile virus (WNV). Although its taxonomic status is still debated, the complex includes species, both globally distributed or with a more limited distribution, morphologically similar and characterised by different physiological and behavioural traits, which affect their ability as vectors. In many European countries, Cx. pipiens and its sibling species Culex torrentium occur in sympatry, exhibiting similar bionomic and morphological characters, but only Cx. pipiens appears to play a vector role in WNV transmission. This species consists of two biotypes, pipiens and molestus, which can interbreed when in sympatry, and their hybrids can act as WNV-bridge vectors, due to intermediate ecological features. Considering the yearly WNV outbreaks since 2008 and given the morphological difficulties in recognising species and biotypes, our aim was to molecularly identify and characterised Cx. pipiens and Cx. torrentium in Italy, using recently developed molecular assays. Culex torrentium was not detected; as in other European countries, the pipiens and molestus biotypes were widely found in sympatry with hybrids in most environments. The UPGMA cluster analysis applied to CQ11 genotypic frequencies mainly revealed two groups of Cx. pipiens populations that differed in ecological features. The high propensity of the molestus biotype to exist in hypogean environments, where the habitat’s physical characteristics hinder and preclude the gene flow, was shown. These results confirmed the CQ11 assay as a reliable diagnostic method, consistent with the ecological and physiological aspects of the populations analysed. Since the assessment of the actual role of three biotypes in the WNV circulation remains a crucial point to be elucidated, this extensive molecular screening of Cx. pipiens populations can provide new insights into the ecology of the species

  3. Ecological Distribution and CQ11 Genetic Structure of Culex pipiens Complex (Diptera: Culicidae) in Italy.

    PubMed

    Di Luca, Marco; Toma, Luciano; Boccolini, Daniela; Severini, Francesco; La Rosa, Giuseppe; Minelli, Giada; Bongiorno, Gioia; Montarsi, Fabrizio; Arnoldi, Daniele; Capelli, Gioia; Rizzoli, Annapaola; Romi, Roberto

    2016-01-01

    Mosquitoes in the Culex pipiens complex are considered to be involved in the transmission of a range of pathogens, including West Nile virus (WNV). Although its taxonomic status is still debated, the complex includes species, both globally distributed or with a more limited distribution, morphologically similar and characterised by different physiological and behavioural traits, which affect their ability as vectors. In many European countries, Cx. pipiens and its sibling species Culex torrentium occur in sympatry, exhibiting similar bionomic and morphological characters, but only Cx. pipiens appears to play a vector role in WNV transmission. This species consists of two biotypes, pipiens and molestus, which can interbreed when in sympatry, and their hybrids can act as WNV-bridge vectors, due to intermediate ecological features. Considering the yearly WNV outbreaks since 2008 and given the morphological difficulties in recognising species and biotypes, our aim was to molecularly identify and characterised Cx. pipiens and Cx. torrentium in Italy, using recently developed molecular assays. Culex torrentium was not detected; as in other European countries, the pipiens and molestus biotypes were widely found in sympatry with hybrids in most environments. The UPGMA cluster analysis applied to CQ11 genotypic frequencies mainly revealed two groups of Cx. pipiens populations that differed in ecological features. The high propensity of the molestus biotype to exist in hypogean environments, where the habitat's physical characteristics hinder and preclude the gene flow, was shown. These results confirmed the CQ11 assay as a reliable diagnostic method, consistent with the ecological and physiological aspects of the populations analysed. Since the assessment of the actual role of three biotypes in the WNV circulation remains a crucial point to be elucidated, this extensive molecular screening of Cx. pipiens populations can provide new insights into the ecology of the species

  4. Reexamination of Culex pipiens Hybridization Zone in the Eastern United States by Ribosomal DNA-Based Single Nucleotide Polymorphism Markers

    PubMed Central

    Huang, Shaoming; Molaei, Goudarz; Andreadis, Theodore G.

    2011-01-01

    Mosquitoes in the Culex pipiens complex are important vectors of several disease-causing pathogens, including West Nile virus. In North America, the complex consists of Cx. pipiens pipiens form pipiens, Cx. pipiens pipiens form molestus, Cx. pipiens quinquefasciatus, and their hybrids that exhibit substantial diversity in physiology, behavior, and geographic range. Hybridization among these mosquitoes is of concern because of potential implications for disease transmission. Currently, several morphological and molecular markers exist for differentiating members of the Cx. pipiens complex; however, these markers have specific limitations. We report here two highly reliable ribosomal DNA-based single nucleotide polymorphism (SNP) markers, CxpG2T and CxpA2d, for detecting Cx. pipiens complex mosquitoes containing Cx. p. quinquefasciatus alleles. Both CxpG2T and CxpA2d contain one allele that is present in all members of the Cx. pipiens complex, and the other allele is specific to Cx. p. quinquefasciatus. Testing of field populations from the eastern United States showed that these two SNP markers are capable of identifying a south to north gradient of Cx. p. quinquefasciatus and hybrids. The northern limit of detection of Cx. p. quinquefasciatus alleles in this study was in Fort Totten, NY (40.79°N), whereas the southern boundary was determined between Atlanta, GA (33.81°N) and Gainesville, FL (29.64°N). CxpG2T and CxpA2d were more accurate than the ACE-2 marker, and they may conceivably provide comparable resolution with microsatellite markers for detecting Cx. p. quinquefasciatus alleles. PMID:21896800

  5. The distribution of potential West Nile virus vectors, Culex pipiens pipiens and Culex pipiens quinquefasciatus (Diptera: Culicidae), in Mexico City

    PubMed Central

    2011-01-01

    Background Culex spp. mosquitoes are considered to be the most important vectors of West Nile virus (WNV) detected in at least 34 species of mosquitoes in the United States. In North America, Culex pipiens pipiens, Culex pipiens quinquefasciatus, and Culex tarsalis are all competent vectors of WNV, which is considered to be enzootic in the United States and has also been detected in equines and birds in many states of Mexico and in humans in Nuevo Leon. There is potential for WNV to be introduced into Mexico City by various means including infected mosquitoes on airplanes, migrating birds, ground transportation and infected humans. Little is known of the geographic distribution of Culex pipiens complex mosquitoes and hybrids in Mexico City. Culex pipiens pipiens preferentially feed on avian hosts; Culex pipiens quinquefasciatus have historically been considered to prefer mammalian hosts; and hybrids of these two species could theoretically serve as bridge vectors to transmit WNV from avian hosts to humans and other mammalian hosts. In order to address the potential of WNV being introduced into Mexico City, we have determined the identity and spatial distribution of Culex pipiens complex mosquitoes and their hybrids. Results Mosquito larvae collected from 103 sites throughout Mexico City during 2004-2005 were identified as Culex, Culiseta or Ochlerotatus by morphological analysis. Within the genus Culex, specimens were further identified as Culex tarsalis or as belonging to the Culex pipiens complex. Members of the Culex pipiens complex were separated by measuring the ratio of the dorsal and ventral arms (DV/D ratio) of the male genitalia and also by using diagnostic primers designed for the Ace.2 gene. Culex pipiens quinquefasciatus was the most abundant form collected. Conclusions Important WNV vectors species, Cx. p. pipiens, Cx. p. quinquefasciatus and Cx. tarsalis, are all present in Mexico City. Hybrids of Cx. p. pipiens and Cx. p. quinquefasciatus were also

  6. IMPLICATIONS OF THE DISCOVERY OF RANA YAVAPAIENSIS IN THE WESTERN GRAND CANYON TO THE CONSERVATION STRATEGY FOR RANA ONCA

    EPA Science Inventory

    The minimum historical range of the relict leopard frog, Rana onca, comprises the drainages of the Virgin and Colorado rivers from the vicinity ofHurricane, Utah, to Black Canyon below Lake Mead, in Nevada and Arizona. Extant populations are known near only the Black Canyon and O...

  7. Seasonal Abundance of Culex tarsalis and Culex pipiens Complex Mosquitoes (Diptera: Culicidae) in California

    PubMed Central

    BARKER, CHRISTOPHER M.; ELDRIDGE, BRUCE F.; REISEN, WILLIAM K.

    2010-01-01

    Large-scale patterns in the seasonal abundance profiles of the arboviral mosquito vectors Culex tarsalis Coquillett and the species of the Culex pipiens complex were described based on a decade of counts from 868 New Jersey light traps located throughout the urban and agricultural areas of California. Mean seasonal abundance profiles varied markedly among hydrologic regions. For all species, abundance increased earlier and declined later in drier, warmer southern regions, but variation could not be explained fully by latitude. The observed patterns may be driven by temperature, availability of larval habitats, and for the Cx. pipiens complex, the taxonomic composition of local populations. PMID:20939368

  8. Venom allergen 5 is Associated With Deltamethrin Resistance in Culex pipiens pallens (Diptera: Culicidae)

    PubMed Central

    Lv, Yuan; Lei, Zhentao; Hong, Shanchao; Wang, Weijie; Zhang, Donghui; Zhou, Dan; Sun, Yan; Ma, Lei; Shen, Bo; Zhu, Changliang

    2015-01-01

    The mosquito, Culex pipiens pallens (L.), is an important vector of encephalitis and filariasis in northern China. The control of these mosquitoes occurs primarily via the use of pyrethroid insecticides, such as deltamethrin. The widespread and improper application of pyrethroid has resulted in the evolution of pyrethroid resistance amongst many mosquito populations, including Cx. pipiens pallens. Previous studies using high-throughput transcriptome sequencing have identified that the venom allergen 5 gene is differentially expressed between deltamethrin-susceptible and deltamethrin-resistant Cx. pipiens pallens. In this study, quantitative real-time polymerase chain reaction analyses revealed that venom allergen 5 was significantly overexpressed in adult females of both deltamethrin-resistant laboratory populations and two field populations. The transcriptional level of venom allergen 5 in the laboratory populations was elevated as the levels of deltamethrin resistance increased. Full-length cDNAs of the venom allergen 5 gene were cloned from Cx. pipiens pallens, and contained an open reading frame of 765 bp, encoding a protein with 254 amino acids. The deduced amino acid sequence shared 100% identity with the ortholog in Culex quinquefasciatus Say. The overexpression of venom allergen 5 decreased the susceptibility of mosquito cells to deltamethrin, while knockdown of this gene by RNAi increased the susceptibility of mosquitoes to deltamethrin. This study provides the first evidence of the association between the venom allergen 5 gene and deltamethrin resistance in mosquitoes. PMID:26335474

  9. RANA CATESBEIANA (AMERICAN BULLFROG) DIET

    EPA Science Inventory

    RANA CATESBELANA (American Bullfrog). DIET. Data were obtained opportunistically
    from 28 adult (M = 14; F = 14) bullftogs collected in April 2001 from the Meadow Valley Wash
    located between the cities of Carp and Elgin, Lincoln County, Nevada, USA (N37'17':WI14'30'). Alth...

  10. The effects of resource type and ratio on competition with Aedes albopictus and Culex pipiens (Diptera:Culicidae).

    PubMed

    Costanzo, K S; Muturi, E J; Lampman, R L; Alto, B W

    2011-01-01

    The introduction of Aedes albopictus (Skuse) in the United States has been associated with declines in abundance of resident mosquito species, presumably because of resource competition, as larvae of Ae. albopictus have been illustrated as superior competitors under certain resource conditions. We evaluated the hypothesis that varying the type and ratio of two food resources (Foxtail grass: American elm) alters the competitive outcome of Ae. albopictus and Culex pipiens (L.). We measured survivorship, development time, size, and adult longevity, and estimated the population growth index (A') of populations raised both alone and in equal number with the interspecific competitor, across five ratios of the two food resources. Competition was asymmetric with Ae. albopictus, the superior competitor across all resource treatments; however, the competitive advantage Ae. albopictus had over Cx. pipiens was reduced as grass became the predominant resource. With elm as the predominant resource, the population growth index (A') for both Ae. albopictus and Cx. pipiens was lower in intraspecific and interspecific competition treatments, respectively. The treatments also impacted adult life history, as life spans of both Ae. albopictus and Cx. pipiens varied when they emerged from larval conditions with different resource and competition treatments. We discuss the possible differences in the two species' efficiencies in exploiting the two resource types. Despite some resource conditions alleviating the competitive effects of Ae. albopictus on Cx. pipiens, competition remained asymmetric; thus, additional mechanisms are likely operating under field conditions when the two species coexist. PMID:21337945

  11. EFFECTS OF LABORATORY ULTRAVIOLET LIGHT AND NATURAL SUNLIGHT ON SURVIVAL AND DEVELOPMENT OF RANA PIPIENS

    EPA Science Inventory

    Changes in solar ultraviolet (UV) radiation have been proposed as a possible factor contributing to seeming increases in hindlimb malformations in anuran amphibians in North America. A primary purpose of this study was to reproduce results from an earlier experiment in which Ran...

  12. CHRONIC RETINOID EXPOSURE IN RANA PIPIENS AND XENOPUS LAEVIS: LIFE-STAGE SPECIFIC SENSITIVITY

    EPA Science Inventory

    Recently, high frequencies of malformations have been reported in amphibians across the United States. It has been suggested that the malformations may be the result of xenobiotic disruption of retinoid signaling...

  13. The pattern of catecholamine response to burst activity in leopard frogs, Rana pipiens.

    PubMed

    Fournier, P A; Nadeau, A; Guderley, H

    1994-07-01

    It is well known that burst activity causes a rapid breakdown of muscle glycogen and extensive accumulation of lactate in frogs. During recovery, it has been shown that lactate is nearly totally recycled into muscle glycogen. Since catecholamines are likely to play some role in the regulation of postexercise repletion of muscle glycogen, the pattern of catecholamine response was assessed in frogs during intense physical activity and the ensuing recovery period. Chronically cannulated frogs were forced to swim until exhaustion, and serial blood samples were taken at regular time intervals for the measurements of catecholamines. The pattern of changes in plasma and muscle lactate and glucose and muscle glycogen during and after burst activity is similar to that reported in previous studies using noncannulated frogs, a result which indicates that the animals recover well from the surgical trauma associated with cannulation. The concentrations of plasma catecholamines in frogs at rest are comparable to those measured in other amphibians, and the levels of plasma epinephrine in resting frogs are much higher than those of norepinephrine. Burst activity causes a marked increase in plasma catecholamines, with higher levels reached by epinephrine. During recovery, the concentration of plasma catecholamines returns to normal within 30 min. Although this pattern of catecholamine response to intense physical activity may be favorable to the repletion of muscle glycogen postexercise, it remains to be clarified how critical the low levels and fast reduction in plasma catecholamines are for optimum glycogen resynthesis. PMID:7926648

  14. Metabolic fate of lactate after vigorous activity in the leopard frog, Rana pipiens.

    PubMed

    Fournier, P A; Guderley, H

    1992-02-01

    Although the ability of isolated frog muscle to synthesize glycogen from lactate has long been known, it has never been demonstrated that this metabolic activity occurs in the intact frog. Our results clearly indicate that lactate glycogenesis in frog muscle occurs to a significant extent in vivo. During recovery from strenuous exercise, most of the lactate accumulated by frogs seems to be recycled into muscle glycogen because the lactate that disappears during recovery could account nearly stoichiometrically for the glycogen that accumulates in muscle. Furthermore, the decrease in body lactate and the increase in muscle glycogen follow corresponding time courses, suggesting a precursor-product relationship between lactate and glycogen. During recovery from intense exercise, hepatectomized and normal frogs have nearly identical extents of lactate elimination and glycogen synthesis. This suggests that muscle is the main tissue responsible for the recycling of lactate into muscle glycogen and that liver plays a negligible role in lactate disposal. The negligible hepatic contribution to lactate recycling results in part from the liver's incapacity to produce glucose from lactate. In support of this proposition, we show that frog liver perfused in vitro is unable to incorporate any detectable labeled lactate into glucose despite its excellent physiological integrity. Changes in dietary status, training state, season at which the experiments were done, exercise status, and composition of the perfusion media (pH, hormonal composition, physiological saline vs. culture medium) did not give rise to lactate gluconeogenesis. Because frog liver contains all the regulatory enzymes of the gluconeogenic pathway, its inability to synthesize glucose from lactate is not due to an absence of pyruvate carboxylase. A limited ability for lactate uptake may explain why frog liver cannot produce glucose from lactate. PMID:1539733

  15. Representation of the visual field in the anterior thalamus of the leopard frog, Rana pipiens.

    PubMed

    Skorina, Laura K; Recktenwald, Eric W; Dudkin, Elizabeth A; Saidel, William M; Gruberg, Edward R

    2016-05-16

    We used physiological and anatomical methods to elucidate how the visual field is represented in the part of the dorsal anterior thalamus of the leopard frog that receives direct retinal projections. We recorded extracellularly while presenting visual stimuli, and characterized a physiologically defined region that encompasses the retinal projections as well as an extended zone beyond them. We probed the area systematically to determine if the zone is organized in a visuotopic map: we found that it is not. We found that units in this region respond only to stimuli in the contralateral half of the visual field, which is similar to what is seen in the dorsal lateral geniculate nucleus in mammals. When we backfilled retinal ganglion cells from application of HRP to the anterior thalamus, we found labeled cells only in those parts of the retina corresponding to the contralateral hemifield, confirming our physiological observations. PMID:27064110

  16. Complex spatial dynamics maintain northern leopard frog (Lithobates pipiens) genetic diversity in a temporally varying landscape

    USGS Publications Warehouse

    Mushet, David M.; Euliss, Ned H.; Chen, Yongjiu; Stockwell, Craig A.

    2013-01-01

    In contrast to most local amphibian populations, northeastern populations of the Northern Leopard Frog (Lithobates pipiens) have displayed uncharacteristically high levels of genetic diversity that have been attributed to large, stable populations. However, this widely distributed species also occurs in areas known for great climatic fluctuations that should be reflected in corresponding fluctuations in population sizes and reduced genetic diversity. To test our hypothesis that Northern Leopard Frog genetic diversity would be reduced in areas subjected to significant climate variability, we examined the genetic diversity of L. pipiens collected from 12 sites within the Prairie Pothole Region of North Dakota. Despite the region's fluctuating climate that includes periods of recurring drought and deluge, we found unexpectedly high levels of genetic diversity approaching that of northeastern populations. Further, genetic structure at a landscape scale was strikingly homogeneous; genetic differentiation estimates (Dest) averaged 0.10 (SD = 0.036) across the six microsatellite loci we studied, and two Bayesian assignment tests (STRUCTURE and BAPS) failed to reveal the development of significant population structure across the 68 km breadth of our study area. These results suggest that L. pipiens in the Prairie Pothole Region consists of a large, panmictic population capable of maintaining high genetic diversity in the face of marked climate variability.

  17. Parasites of the mink frog (rana septentrionalis) from minnesota, U.S.A.

    USGS Publications Warehouse

    Schotthoefer, A.M.; Bolek, M.G.; Cole, R.A.; Beasley, V.R.

    2009-01-01

    Twenty-two mink frogs, Rana septentrionalis, collected from two locations in Minnesota, United States, were examined for helminth and protozoan blood parasites in July 1999. A total of 16 parasite taxa were recovered including 5 larval digenean trematodes, 7 adult digenean trematodes, 3 nematodes, and I Trypanosorna species. Infracommunities were dominated by the digeneans in terms of richness and abundance. In particular, echinostomatid metacercariae in the kidneys of frogs were the most common parasites found, infecting 100% of the frogs and consisting of about 90% of all helminth individuals recovered. Gorgodera amplicava, Gorgoderina multilohata, Haernaroloechus pan'iplexus, Haernatoloechus breviplexus, Cosnwcercoides dukae, and Oswaldocruzia pipiens represent new host records. The survey presented here represents the second known helminth survey of mink frogs conducted in North America. A summary of metazoan parasites reported from mink frogs is included.

  18. Patterns of Cranial Development in Larval Rana macrocnemis: Chondrocranial Size and Shape Relationship With Pelophylax bedriagae (Anura: Ranidae).

    PubMed

    Yildirim, Elıf; Kaya, Uğur

    2016-06-01

    Notwithstanding the abundance of amphibians, there are few descriptions about ranid cranial development. Herein, larval chondrocranial development of Uludağ frog, Rana macrocnemis (Boulenger, 1885), is described on cleared and double-stained specimens. Descriptions are related with the ontogeny of the chondrocranium and osteogenesis of the cranial skeleton. The larval chondrocranial development of R. macrocnemis is compared to those of Rana and Pelophylax larvae (Pelophylax bedriagae, Rana pipiens, R. palustris, R. sphenocephala, R. catesbeiana, R. clamitans and R. sylvatica). In R. macrocnemis, the first bones to ossify are the parasphenoid and exoccipital (Stage 33), followed by the frontoparietal and prootic (stages 35 and 40, respectively). The major reconstruction of the chondrocranium begins at Stage 41. The ossification sequence of R. macrocnemis is distinguished from other ranids. Adult cranial osteology of R. macrocnemis is compared to that of P. bedriagae. Osteologically, R. macrocnemis is different from P. bedriagae by the shape and size of the vomer and number of teeth. Additionally, geometric morphometric methods are used to analyze chondrocranial size and shape changes of ranid larva of R. macrocnemis and P. bedriagae. Anat Rec, 299:711-721, 2016. © 2016 Wiley Periodicals, Inc. PMID:26950267

  19. Efficient induction of spawning of Northern leopard frogs (Lithobates pipiens) during and outside the natural breeding season

    PubMed Central

    2013-01-01

    Background Amphibian declines are now recognized globally. It is also well known that many anurans do not reproduce easily in captivity, especially when held over long periods, or if they require hibernation before breeding. A simple method to induce spawning and subsequent development of large numbers of healthy tadpoles is therefore required to meet research and conservation goals. Methods The method is based on simultaneous injection of both female and male leopard frogs, Lithobates pipiens (formerly called Rana pipiens) with a cocktail of a gonadotropin-releasing hormone agonist (GnRH-A) and a dopamine antagonist. We call this the AMPHIPLEX method, which is derived from the combination of the words amphibian and amplexus. Following injection, the animals are thereby induced, and perform amplexus and natural fertilization under captive conditions. Results We tested combinations of a GnRH agonist with 2 different dopamine antagonists in L. pipiens in the breeding season. The combination of des-Gly10, D-Ala6, Pro-NHEt9-GnRH (0.4 micrograms/g body weight; GnRH-A) with metoclopramide hydrochloride (10 micrograms/g body weight; MET) or domperidone (DOM) were equally effective, producing 89% and 88% successful spawning, respectively. This yielded more than 44,000 eggs for the 16/18 females that ovulated in the GnRH-A+MET group, and more than 39,000 eggs for the 15/17 females that ovulated in the GnRH-A+DOM group. We further tested the GnRH-A+MET in frogs collected in the wild in late autumn and hibernated for a short period under laboratory conditions, and report a low spawning success (43%). However, GnRH-A priming 24 hours prior to injections of the GnRH-A+MET cocktail in animals hibernated for 5–6 weeks produced out-of-season spawning (89%) and fertilization (85%) comparable to those we observed for in-season spawning. Assessment of age and weight at metamorphosis indicated that L. pipiens tadpoles resulting from out-of-season spawning grew normally and

  20. The Culex pipiens fatigans problem in South-East Asia

    PubMed Central

    Singh, D.

    1967-01-01

    In South-East Asia in recent years urbanization has proceeded rapidly. Because of limited financial resources it has seldom been possible to provide sufficient sanitation; this has led to conditions favourable to the breeding of Culex pipiens fatigans. The density of C. p. fatigans is higher in urban areas than in rural ones. Differences in infection and infectivity rates in C. p. fatigans seem to be due to differences in infection rates in the human population rather than to differences in the life-span of the mosquito. Precipitin tests have shown that the species feeds predominantly on man. It breeds in almost any type of stagnant water with organic contamination. Filariasis problems in India have been classified according to the duration of established transmission. PMID:5300060

  1. EVIDENCE FOR PHYLOGENETICALLY DISTINCT LEOPARD FROGS (RANA ONCA) FROM THE BORDER REGION OF NEVADA, UTAH, ARIZONA

    EPA Science Inventory


    Remnant populations of leopard frogs exist within the Virgin River drainage and adjacent portions of the Colorado River (Black Canyon) in northwestern Arizona and southern Nevada. These populations either represent the reportedly extinct taxa Rana onca or northern, disjunct R...

  2. Identification of QTLs Conferring Resistance to Deltamethrin in Culex pipiens pallens

    PubMed Central

    Zhong, Daibin; Shen, Bo; Zhang, Donghui; Guo, Qin; Wang, Weijie; Yu, Jing; Lv, Yuan; Lei, Zhentao; Ma, Kai; Ma, Lei; Zhu, Changliang; Yan, Guiyun

    2015-01-01

    Culex pipiens pallens is the most abundant Culex mosquito species in northern China and is an important vector of bancroftian filariasis and West Nile virus. Deltamethrin is an insecticide that is widely used for mosquito control, however resistance to this and other insecticides has become a major challenge in the control of vector-borne diseases that appear to be inherited quantitatively. Furthermore, the genetic basis of insecticide resistance remains poorly understood. In this study, quantitative trait loci (QTL) mapping of resistance to deltamethrin was conducted in F2 intercross segregation populations using bulked segregation analysis (BSA) and amplified fragment length polymorphism markers (AFLP) in Culex pipiens pallens. A genetic linkage map covering 381 cM was constructed and a total of seven QTL responsible for resistance to deltamethrin were detected by composite interval mapping (CIM), which explained 95% of the phenotypic variance. The major QTL in linkage group 2 accounted for 62% of the variance and is worthy of further study. 12 AFLP markers in the map were cloned and the genomic locations of these marker sequences were determined by applying the Basic Local Alignment Search Tool (BLAST) tool to the genome sequence of the closely related Culex quinquefasciatus. Our results suggest that resistance to deltamethrin is a quantitative trait under the control of a major QTL in Culex pipiens pallens. Cloning of related AFLP markers confirm the potential utility for anchoring the genetic map to the physical map. The results provide insight into the genetic architecture of the trait. PMID:26484540

  3. Repeated bouts of dehydration deplete nutrient reserves and reduce egg production in the mosquito Culex pipiens

    PubMed Central

    Benoit, Joshua B.; Patrick, Kevin R.; Desai, Karina; Hardesty, Jeffrey J.; Krause, Tyler B.; Denlinger, David L.

    2010-01-01

    In this study of the mosquito, Culex pipiens, we examined the impact of multiple bouts of dehydration and rehydration on survival, depletion of metabolic reserves and egg production in both non-diapausing and diapausing females. Mosquitoes provided with access to sugar during rehydration survived longer than those allowed to rehydrate without sugar, and their survival was similar to that of mosquitoes of the same age that were not dehydrated. Among mosquitoes not provided with sugar, each dehydration bout reduced the mosquito's dry mass – an effect likely to be due to the utilization of carbohydrates and lipid reserves. The toll on glycogen and lipid reserves is likely to be especially costly for diapausing mosquitoes that are dependent on these stored reserves for winter survival. Egg production in both non-diapausing and post-diapausing C. pipiens was also reduced in response to multiple bouts of dehydration. Although egg quality was not compromised, the number of eggs produced was reduced. Both non-diapausing and diapausing females can compensate for the nutrient loss due to dehydration by sugar feeding but the opportunity to feed on sugar is likely to be rarely available in the overwintering habitat of diapausing females, thus the impact of dehydration may be especially pronounced in overwintering populations of C. pipiens. PMID:20675546

  4. Evolutionary genomics of Culex pipiens: global and local adaptations associated with climate, life-history traits and anthropogenic factors

    PubMed Central

    Asgharian, Hosseinali; Chang, Peter L.; Lysenkov, Sergey; Scobeyeva, Victoria A.; Reisen, William K.; Nuzhdin, Sergey V.

    2015-01-01

    We present the first genome-wide study of recent evolution in Culex pipiens species complex focusing on the genomic extent, functional targets and likely causes of global and local adaptations. We resequenced pooled samples of six populations of C. pipiens and two populations of the outgroup Culex torrentium. We used principal component analysis to systematically study differential natural selection across populations and developed a phylogenetic scanning method to analyse admixture without haplotype data. We found evidence for the prominent role of geographical distribution in shaping population structure and specifying patterns of genomic selection. Multiple adaptive events, involving genes implicated with autogeny, diapause and insecticide resistance were limited to specific populations. We estimate that about 5–20% of the genes (including several histone genes) and almost half of the annotated pathways were undergoing selective sweeps in each population. The high occurrence of sweeps in non-genic regions and in chromatin remodelling genes indicated the adaptive importance of gene expression changes. We hypothesize that global adaptive processes in the C. pipiens complex are potentially associated with South to North range expansion, requiring adjustments in chromatin conformation. Strong local signature of adaptation and emergence of hybrid bridge vectors necessitate genomic assessment of populations before specifying control agents. PMID:26085592

  5. Evolutionary genomics of Culex pipiens: global and local adaptations associated with climate, life-history traits and anthropogenic factors.

    PubMed

    Asgharian, Hosseinali; Chang, Peter L; Lysenkov, Sergey; Scobeyeva, Victoria A; Reisen, William K; Nuzhdin, Sergey V

    2015-07-01

    We present the first genome-wide study of recent evolution in Culex pipiens species complex focusing on the genomic extent, functional targets and likely causes of global and local adaptations. We resequenced pooled samples of six populations of C. pipiens and two populations of the outgroup Culex torrentium. We used principal component analysis to systematically study differential natural selection across populations and developed a phylogenetic scanning method to analyse admixture without haplotype data. We found evidence for the prominent role of geographical distribution in shaping population structure and specifying patterns of genomic selection. Multiple adaptive events, involving genes implicated with autogeny, diapause and insecticide resistance were limited to specific populations. We estimate that about 5-20% of the genes (including several histone genes) and almost half of the annotated pathways were undergoing selective sweeps in each population. The high occurrence of sweeps in non-genic regions and in chromatin remodelling genes indicated the adaptive importance of gene expression changes. We hypothesize that global adaptive processes in the C. pipiens complex are potentially associated with South to North range expansion, requiring adjustments in chromatin conformation. Strong local signature of adaptation and emergence of hybrid bridge vectors necessitate genomic assessment of populations before specifying control agents. PMID:26085592

  6. The Culex pipiens complex in the Mississippi River basin: identification, distribution, and bloodmeal hosts.

    PubMed

    Savage, Harry M; Kothera, Linda

    2012-12-01

    Members of the Culex pipiens complex are the primary vectors of St. Louis encephalitis virus and West Nile virus in the Mississippi River basin (MRB). The Cx. pipiens complex in the MRB is composed of 4 taxa: Cx. p. pipiens form pipiens, Cx. p. quinquefasciatus, hybrids between Cx. p. pipiens f. pipiens and Cx. p. quinquefasciatus, and Cx. p. pipiens form molestus. Three studies on bloodmeal hosts with large sample sizes have been conducted on members of the Cx. pipiens complex in the MRB including 1 each on Cx. p. quinquefasciatus from Louisiana, Cx. p. pipiens-quinquefasciatus hybrids from Tennessee, and Cx. p. pipiens from Illinois. The top 8 bloodmeal hosts from each of the 3 sites accounted for 68-92% of bloodmeals. Only 14 species accounted for the top 8 bloodmeal hosts at each of the 3 sites. The most often utilized bloodmeal hosts for members of the Culex pipiens complex within the MRB are the American robin, Northern cardinal, human, raccoon, common grackle, house sparrow, mourning dove, dog, Northern mockingbird, blue jay, opossum, domestic horse, house finch and European starling. Human feeding varied widely among sites from 1% to 15.7% of bloodmeals. The proportion of bloodmeals taken on humans is an important epidemiological variable and future studies are needed to define the primary genetic and environmental factors that influence host utilization by members of the Cx. pipiens complex. PMID:23401948

  7. Induction of photolyase activity in wood frog (Rana sylvatica) embryos.

    PubMed

    Smith, M A; Kapron, C M; Berrill, M

    2000-10-01

    Rising ultraviolet-B (UVB, 280-320 nm) radiation has been proposed as a factor which may explain nonnormal amphibian population declines. Accordingly research has been directed toward estimating the photolyase activity of several amphibian species in order to predict a species' resilience to UV damage. Unfortunately, in spite of published research which demonstrated that the activity of one of the principal photorepair enzymes, photolyase, can be induced, these estimates did not address the potential for in vivo induction by environmental factors present in situ. We show here that wood frog (Rana sylvatica) embryos exposed to periods of ambient solar radiation (1) displayed significantly different photolyase activities from embryos exposed to equivalent periods of dark; and (2) were positively correlated with the UVB fluence received in vivo. Such results suggest that previous conclusions regarding the relationship between photorepair and population decline must be reevaluated. Estimating amphibian photorepair is a complicated process, and caution must be exercised when interpreting such data. PMID:11045732

  8. Effects of predatory fish on survival and behavior of larval gopher frogs (Rana capito) and Southern Leopard Frogs (Rana sphenocephala)

    USGS Publications Warehouse

    Gregoire, D.R.; Gunzburger, M.S.

    2008-01-01

    Southern Leopard Frogs, Rana sphenocephala, are habitat generalists occurring in virtually all freshwater habitats within their geographic range, whereas Gopher Frogs, Rana capito, typically breed in ponds that do not normally contain fish. To evaluate the potential for predation by fish to influence the distribution of these species, we conducted a randomized factorial experiment. We examined the survival rate and behavior of tadpoles when exposed to Warmouth Sunfish, Lepomis gulosus, Banded Sunfish, Enneacanthus obesus, and Eastern Mosquitofish, Gambusia holbrooki. We also conducted a choice experiment to examine the survival rate of the two species of tadpoles when a predator is given a choice of both species simultaneously. Lepomis gulosus consumed the most tadpoles and ate significantly more tadpoles of R. capito than R. sphenocephala. Gambusia holbrooki injured the most tadpoles, especially R. capito. Enneacanthus obesus did not have an effect on behavior or survival of either anuran species. Tadpoles of both anurans increased hiding when in the presence of L. gulosus and G. holbrooki, but a greater proportion of R. capito hid than did R. sphenocephala. Our results suggest that R. capito are more vulnerable to predation by fish than are R. sphenocephala. The introduction of fish may play a role in population declines of certain anurans breeding in normally fish-free wetlands, and even small fish, such as mosquitofish, may have significant negative effects on the tadpoles of R. capito. Copyright 2008 Society for the Study or Amphibians and Reptiles.

  9. Body size affects the predatory interactions between introduced American Bullfrogs (Rana catesbeiana) and native anurans in China: An experimental study

    USGS Publications Warehouse

    Wang, Y.; Guo, Z.; Pearl, C.A.; Li, Y.

    2007-01-01

    Introduced American Bullfrogs (Rana catesbeiana) have established breeding populations in several provinces in China since their introduction in 1959. Although Bullfrogs are viewed as a potentially important predator of Chinese native anurans, their impacts in the field are difficult to quantify. We used two experiments to examine factors likely to mediate Bullfrog predation on native anurans. First, we examined effects of Bullfrog size and sex on daily consumption of a common Chinese native (Rana limnocharis). Second, we examined whether Bullfrogs consumed similar proportions of four Chinese natives: Black-Spotted Pond Frog (Rana nigromaculata), Green Pond Frog (Rana plancyi plancyi), Rice Frog (R. limnocharis), and Zhoushan Toad (Bufo bufo gargarizans). We found that larger Rana catesbeiana consumed more R. limnocharis per day than did smaller R. catesbeiana, and that daily consumption of R. limnocharis was positively related to R. catesbeiana body size. When provided with adults of four anurans that differed significantly in body size, R. catesbeiana consumed more individuals of the smallest species (R. limnocharis). However, when provided with similarly sized juveniles of the same four species, R. catesbeiana did not consume any species more than expected by chance. Our results suggest that body size plays an important role in the predatory interactions between R. catesbeiana and Chinese native anurans and that, other things being equal, smaller species and individuals are at greater risk of predation by R. catesbeiana. Copyright 2007 Society for the Study of Amphibians and Reptiles.

  10. Molecular Ecology of Pyrethroid Knockdown Resistance in Culex pipiens pallens Mosquitoes

    PubMed Central

    Zhang, Donghui; Shi, Linna; Zhou, Guofa; Gong, Maoqing; Zhou, Huayun; Sun, Yan; Ma, Lei; He, Ji; Hong, Shanchao; Zhou, Dan; Xiong, Chunrong; Chen, Chen; Zou, Ping; Zhu, Changliang; Yan, Guiyun

    2010-01-01

    Pyrethroid insecticides have been extensively used in China and worldwide for public health pest control. Accurate resistance monitoring is essential to guide the rational use of insecticides and resistance management. Here we examined the nucleotide diversity of the para-sodium channel gene, which confers knockdown resistance (kdr) in Culex pipiens pallens mosquitoes in China. The sequence analysis of the para-sodium channel gene identified L1014F and L1014S mutations. We developed and validated allele-specific PCR and the real-time TaqMan methods for resistance diagnosis. The real-time TaqMan method is more superior to the allele-specific PCR method as evidenced by higher amplification rate and better sensitivity and specificity. Significant positive correlation between kdr allele frequency and bioassay-based resistance phenotype demonstrates that the frequency of L1014F and L1014S mutations in the kdr gene can be used as a molecular marker for deltamethrin resistance monitoring in natural Cx. pipiens pallens populations in the East China region. The laboratory selection experiment found that L1014F mutation frequency, but not L1014S mutation, responded to deltamethrin selection, suggesting that the L1014F mutation is the key mutation conferring resistance to deltamethrin. High L1014F mutation frequency detected in six populations of Cx. pipens pallens suggests high prevalence of pyrethroid resistance in Eastern China, calling for further surveys to map the resistance in China and for investigating alternative mosquito control strategies. PMID:20657783

  11. Molecular ecology of pyrethroid knockdown resistance in Culex pipiens pallens mosquitoes.

    PubMed

    Chen, Lin; Zhong, Daibin; Zhang, Donghui; Shi, Linna; Zhou, Guofa; Gong, Maoqing; Zhou, Huayun; Sun, Yan; Ma, Lei; He, Ji; Hong, Shanchao; Zhou, Dan; Xiong, Chunrong; Chen, Chen; Zou, Ping; Zhu, Changliang; Yan, Guiyun

    2010-01-01

    Pyrethroid insecticides have been extensively used in China and worldwide for public health pest control. Accurate resistance monitoring is essential to guide the rational use of insecticides and resistance management. Here we examined the nucleotide diversity of the para-sodium channel gene, which confers knockdown resistance (kdr) in Culex pipiens pallens mosquitoes in China. The sequence analysis of the para-sodium channel gene identified L1014F and L1014S mutations. We developed and validated allele-specific PCR and the real-time TaqMan methods for resistance diagnosis. The real-time TaqMan method is more superior to the allele-specific PCR method as evidenced by higher amplification rate and better sensitivity and specificity. Significant positive correlation between kdr allele frequency and bioassay-based resistance phenotype demonstrates that the frequency of L1014F and L1014S mutations in the kdr gene can be used as a molecular marker for deltamethrin resistance monitoring in natural Cx. pipiens pallens populations in the East China region. The laboratory selection experiment found that L1014F mutation frequency, but not L1014S mutation, responded to deltamethrin selection, suggesting that the L1014F mutation is the key mutation conferring resistance to deltamethrin. High L1014F mutation frequency detected in six populations of Cx. pipens pallens suggests high prevalence of pyrethroid resistance in Eastern China, calling for further surveys to map the resistance in China and for investigating alternative mosquito control strategies. PMID:20657783

  12. [State resistance of the mosquito Culex pipiens towards temephos central Morocco].

    PubMed

    El Ouali Lalami, A; El-Akhal, F; El Amri, N; Maniar, S; Faraj, C

    2014-08-01

    In Morocco, Culex pipiens plays a role in the high annoyance experienced by most urban cities, suburban and rural areas, especially since it was strongly suspected as the most likely vector in the transmission of West Nile virus epidemics that have hit Morocco in 1996. Chemical insecticides are generally the way in which they use the programs against harmful mosquitoes and disease vectors. However, the repeated and excessive use of these products regularly led to the emergence of the phenomenon of insect resistance. At the center of Morocco, information on the susceptibility or resistance to insecticides in mosquitoes (larvae and adults) vectors of diseases or pests, are almost nonexistent. This article reports the results of studies conducted between 2007 and 2010 with sensitivity tests WHO on larvae local populations of Culex pipiens collected in three lodging in the city of Fez, towards the insecticide mostly used by hygienic services: temephos. Five concentrations of insecticide (0.0025 mg/l, 0.005 mg/l, 0.0125 mg/l, 0.025 mg/l, 0.0625 mg/l) in addition to control, were used to determine the LC50 and LC 90 of Culex pipiens species towards temephos. Sensitivity tests were carried out at the entomology unit and monitoring of insect sensitivity towards insecticides installed at the Regional Diagnostic Laboratory Epidemiological and Environmental Hygiene (LRDEHM), Fez, under the Regional Directorate of Health in Fes Boulemane Region. The LC50 and LC90, concentrations corresponding to 50 and 90% mortality were determined graphically, by the linear relationship between the decimal logarithm of insecticide concentrations (x-axis) and the percentage of mortality transformed into probit values (ordinate) on logarithmic gausso paper. Resistance rates were determined on the basis of the sensitivity of a reference strain (S-Lab). The bioassay results affirmed the presence of resistance in larvae Culex pipiens towards temephos and that this species has also equally

  13. Macrocyclops albidus (Copepoda: cyclopidae) for the Biocontrol of Aedes albopictus and Culex pipiens in Italy.

    PubMed

    Veronesi, Rodolfo; Carrieri, Marco; Maccagnani, Bettina; Maini, Stefano; Bellini, Romeo

    2015-03-01

    The aim of our study was to assess the potential of Macrocyclops albidus as a biological control agent against the 1st and 2nd instars of Culex pipiens and Aedes albopictus. Under laboratory conditions of prey saturation (50 1st instars/copepod), an average of 58.98% of Cx. pipiens and 54.99% of Ae. albopictus larvae were killed by 1 copepod in 24 h. Trials run in big drums containing 200 liters of water showed that the M. albidus population, inoculated in April, efficiently controlled the mosquito population for the entire season. The predator was particularly effective against Ae. albopictus, as only 2 larvae of this species were found in the treated drums, compared to 814 larvae in untreated control drums throughout the study period. No difference was observed in the control efficacy between the 2 initial densities of copepods used. The reduction in Ae. albopictus density in the drums with 100 and 500 M. albidus with respect to the control drums was 99.90 ± 0.35% and 100.0 ± 0.0%, respectively. For Cx. pipiens, the reduction in density was 88.69 ± 13.51% and 84.65% in drums inoculated with 100 and 500 copepods, respectively. Macrocyclops albidus populations survived through the winter and continued to keep the mosquito population under control during the 2008 season. The M. albidus population developed very well both in drums placed in sunny and shaded areas and proved to be tolerant to both high (summer) and low (winter) temperatures. Trials performed on M. albidus survival in catch basins showed that after a few weeks, the copepod population dramatically decreased and subsequently disappeared. The main problem for copepod survival in catch basins seemed to be the low oxygen tension and accumulation of toxic substances, rather than copepods being flushed out in heavy rainfall episodes. During the period when copepods were present, they maintained the mosquito population under control; their partial disappearance from the catch basins, however, would

  14. PARTIAL LIFE-CYCLE TOXICITY AND BIOCONCENTRATION MODELLING OF PERFLUOROOCTANE SULFONATE (PFOS) IN THE NORTHERN LEOPARD FROG (RANA PIPIENS)

    EPA Science Inventory

    A number of recent monitoring studies have demonstrated elevated concentrations of perfluorooctane sulfonate (PFOS) in humans and wildlife throughout the world. Although no longer manufactured in the U.S., the global distribution and relative persistence of PFOS indicates a need ...

  15. The affinity and activity of compounds related to nicotine on the rectus abdominis muscle of the frog (Rana pipiens)

    PubMed Central

    Barlow, R. B.; Thompson, G. M.

    1969-01-01

    1. Series of pyridylalkyl- and substituted phenylalkyl-trimethylammonium salts, triethylammonium salts, diethylamines and di-n-propylamines have been made. The substituents in the benzene ring were nitro, chloro, bromo, methoxy, hydroxy and amino groups and the alkyl residues had one, two, or three methylene groups separating the aromatic nucleus from the cationic head. 2. Most of the trimethylammonium compounds caused a contracture of the frog rectus muscle, but some were partial agonists and a few were antagonists. The di-n-propylamines were all antagonists, as were most of the diethylamines and triethylammonium compounds, though some of these were partial agonists and a few triethylammonium compounds were agonists. The affinities of the antagonists and partial agonists for the receptors stimulated by β-pyridylmethyltrimethylammonium (and by nicotine) were measured. The equipotent molar ratios of all the agonists were measured relative to β-pyridylmethyltrimethylammonium. 3. The dissociation constants of the pyridylmethyldiethylamines and substituted benzyldiethylamines were measured. The effects of substituents on the pKa of benzyldiethylamine were similar to their effects on the pKa of aniline, though there were differences with some of the o-substituted compounds, which could be attributed to internal hydrogen-bond formation. 4. There is no obvious correlation between the effects of a substituent on the pKa of benzyldiethylamine and its effects on affinity. Although increasing the size of the cationic group usually increased affinity, it did not always do so. The compounds with the highest affinity, p-hydroxybenzyldiethylamine (log K, 5·90) had about half the affinity of (+)-tubocurarine (log K, 6·11), but the triethylammonium analogue (log K, 4·17) had only about one-fiftieth of the affinity of the tertiary base. The binding of the drug to the receptor appears to involve many factors which include the size of the groups as well as their electron-releasing or withdrawing nature and other properties, such as their polar and lipophilic or lipophobic character. 5. There is no obvious correlation between the effects of a substituent on the affinity of the diethylamino or triethylammonium compounds and its effects on the activity of the trimethylammonium analogue. The most active compounds contain hydroxy- and amino-, phenyl or β-pyridyl groups, m-hydroxyphenyl-propyltrimethylammonium being about 50 times as active as nicotine, but the corresponding diethylamino or triethylammonium compounds do not have high affinity. There does not seem necessarily to be an inverse relationship between activity and affinity, however, because some m-nitro and m-chloro trimethylammonium compounds have considerable activity and the analogous triethylammonium compounds have considerable affinity. 6. It is suggested that ability to activate these receptors is associated with the presence of substituents which can interact with water molecules which may be involved in the action of the drug at the receptor. PMID:5348464

  16. Repellent activity of five essential oils against Culex pipiens.

    PubMed

    Erler, F; Ulug, I; Yalcinkaya, B

    2006-12-01

    Essential oils extracted from the seeds of anise (Pimpinella anisum), dried fruits of eucalyptus (Eucalyptus camaldulensis), dried foliage of mint (Mentha piperita) and basil (Ocimum basilicum) and fresh foliage of laurel (Laurus nobilis) were tested for their repellency against the adult females of Culex pipiens. All essential oils showed repellency in varying degrees, eucalyptus, basil and anise being the most active. PMID:16890387

  17. Multi-character approach reveals a discordant pattern of phenotypic variation during ontogeny in Culex pipiens biotypes (Diptera: Culicidae).

    PubMed

    Krtinić, B; Ludoški, J; Milankov, V

    2015-02-01

    Culex (Culex) pipiens s.l. (Diptera: Culicidae) comprises two distinct biotypes, pipiens ('rural') and molestus ('urban'), both of which are thought to have differing capacities due to different host preferences. To better understand West Nile encephalitis epidemiology and improve risk assessment, local distinction between these forms is essential. This study assesses phenotypic variation at larval and adult stages of 'urban' and 'rural' biotypes of the species by complementary use of meristic, univariate and multivariate traits analyzed by traditional and geometric morphometrics. Third- and fourth-instar larvae from a broad area of the city of Novi Sad (Serbia) were collected and reared in the laboratory. After adult eclosion, the sex of each larva was recorded based on the sex of the corresponding adult. Examination of the association between variations of larval traits revealed contrasting variations regarding pecten spines vs. siphonal size and siphonal shape in the 'rural' biotype. Siphons of larvae collected in marshes and forest ecosystems outside urban areas were found to be the largest, but possessed the smallest number of pecten spines. In addition, statistically significant female-biased sexual dimorphism was observed in siphonal size, wing size and wing shape. Finally, we propose that an integrative approach is essential in delimitation of Cx. pipiens s.l. biotypes, since their differentiation was not possible based solely on larval and adult traits. Our findings shed light on the phenotypic plasticity important for population persistence in the changing environment of these medically important taxa. PMID:25424880

  18. The identity of Rana lutea Molina, 1782 (Amphibia, Anura).

    PubMed

    Ferraro, Daiana P; Lavilla, Esteban O

    2013-01-01

    Rana lutea is one of the two anuran species described in 1782 by the Chilean born priest and naturalist Juan Ignacio Molina, and its nomenclatural history is confusing. After a detailed revision of the pertinent literature, we demonstrate that Bufo thaul Schneider, 1799, is a junior objective synonym of Rana lutea Molina, 1782, but based on the usage of both nomina, we revert the precedence and consider Rana lutea Molina, 1782 as a nomen oblitum whereas Bufo thaul Schneider, 1799 is a nomen protectum. Thus, the authorship of the species currently known as Pleurodema thaul (Lesson, 1826) changes to Pleurodema thaul (Schneider, 1799). PMID:24614468

  19. Evaluation of some medicinal plants in controlling Culex pipiens.

    PubMed

    Abdel-Sattar, Essami; Zaitoon, Ahmed; El Sayed, Abeer M; Bakhashwain, A A

    2014-12-01

    The study evaluated the efficacy of methanolic extracts of Ruta chalepensis (Rutaceae), Withania somnifera (Solanaceae), Cleome paradoxa (Capparaceae) and Heliotropium longiflorum (Boraginaceae) aerial parts against Culex pipiens larvae. Different concentrations (100-500 ppm) of the methanolic extracts of the plants were tested towards larval mortality and development of C. pipiens separately. Larval mortalities were counted at 2, 4 & 10 days after treatment. Egg hatchability was determined at 4 & 7 days after treatment. Successful pupation and adult emergence percentage were recorded daily till all control adults emerged. All plants extracts exhibited variable activities. The greatest effect was with R. chalepensis which showed acute (2 days) and chronic (10 days) LC50 of 132.6 & 96.56 ppm, respectively. Larval mortality up to 84.47% &85.53%, were observed with C. paradoxa and R. Chalepensis respectively. Egg hatch was significantly reduced about equal with R. chalepensis and W. somnifera extracts. Concentration levels of C. paradoxa (≥200 ppm) and H. longiflorum (>400 ppm) showed significant hindrance to the larval development and reduction to resulting pupae and adults. Drastic development retardation was shown with extract of C. paradoxa leaves (300 ppm), but only 15.3% & 5.6% of larvae reach pupal and adult stages respectively. The larvicidal activity of methanolic extracts of R. chalepensis, W somnifera, C. paradoxa, H. longiflorum proved to be effective against C. pipiens larvae without any human or animal risk. PMID:25643518

  20. Decapitation Improves Detection of Wolbachia pipientis (Rickettsiales: Anaplasmataceae) in Culex pipiens (Diptera: Culicidae) Mosquitoes by the Polymerase Chain Reaction

    PubMed Central

    BECKMANN, J. F.; FALLON, A. M.

    2013-01-01

    Polymerase chain reaction (PCR) is often used to detect microorganisms, pathogens, or both, including the reproductive parasite Wolbachia pipientis (Rickettsiales: Anaplasmataceae), in mosquitoes. Natural populations of Culex pipiens L. (Diptera: Culicidae) mosquitoes are infected with one or more strains of W. pipientis, and crosses between mosquitoes harboring different Wolbachia strains provide one of the best-known examples of cytoplasmic incompatibililty (CI). When we used PCR to monitor Wolbachia in the Buckeye strain of Culex pipiens, and in a Wolbachia-cured sister colony obtained by tetracycline treatment, we noted false negative PCR reactions with DNA samples from infected mosquitoes; these results were inconsistent with direct microscopic observation of Wolbachia-like particles in gonads dissected from mosquitoes in the same population. Assays with diluted template often improved detection of positive samples, suggesting that DNA prepared from whole mosquitoes contained an inhibitor of the PCR reaction. We reconciled discrepancies between PCR and microscopy by systematic measurement of the PCR reaction in the presence of an internal standard. Mosquito decapitation before DNA extraction restored the reliability of the PCR reaction, allowing accurate determination of Wolbachia infection status in infected and tetracycline-cured mosquito populations, consistent with microscopic examination. Using PCR primers based on the Tr1 gene, we confirmed that the Wolbachia infection in the Buckeye strain of Culex pipiens belongs to the genotype designated wPip1. Finally, to explore more widely the distribution of PCR inhibitors, we demonstrated that DNA isolated from the cricket, Acheta domesticus (L.); the beetle, Tenebrio molitor L.; the honey bee, Apis mellifera L.; and the mosquito, Anopheles punctipennis Say also contained PCR inhibitors. These results underscore the importance of measuring the presence of inhibitors in PCR templates by using a known positive

  1. Comparative transcriptome analyses of deltamethrin-susceptible and -resistant Culex pipiens pallens by RNA-seq.

    PubMed

    Lv, Yuan; Wang, Weijie; Hong, Shanchao; Lei, Zhentao; Fang, Fujin; Guo, Qin; Hu, Shengli; Tian, Mengmeng; Liu, Bingqian; Zhang, Donghui; Sun, Yan; Ma, Lei; Shen, Bo; Zhou, Dan; Zhu, Changliang

    2016-02-01

    The widespread and improper use of pyrethroid insecticides, such as deltamethrin, has resulted in the evolution of resistance in many mosquito species, including Culex pipiens pallens. With the development of high-throughput sequencing, it is possible to massively screen pyrethroid resistance-associated gene. In this study, we used Illumina-Solexa transcriptome sequencing to identify genes that are expressed differently in deltamethrin-susceptible and -resistant strains of Culex pipiens pallens as a critical knowledge base for further studies. A total of 4,961,197,620 base pairs and 55,124,418 reads were sequenced, mapped to the Culex quinquefasciatus genome and assembled into 17,679 known genes. We recorded 1826 significantly differentially expressed genes (DEGs). Among them, 1078 genes were up-regulated and 748 genes were down-regulated in the deltamethrin-resistant strain compared to -susceptible strain. These DEGs contained cytochrome P450 s, cuticle proteins, UDP-glucuronosyltransferases, lipases, serine proteases, heat shock proteins, esterases and others. Among the 1826 DEGs, we found that the transcriptional levels of CYP6AA9 in the laboratory populations was elevated as the levels of deltamethrin resistance increased. Moreover, the expression levels of the CYP6AA9 were significantly higher in the resistant strains than the susceptible strains in three different field populations. We further confirmed the association between the CYP6AA9 gene and deltamethrin resistance in mosquitoes by RNA interfering (RNAi). Altogether, we explored massive potential pyrethroid resistance-associated genes and demonstrated that CYP6AA9 participated in the pyrethroid resistance in mosquitoes. PMID:26377942

  2. Global Perspective on the Culex pipiens Complex in the 21st Century I

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The Culex pipiens complex, including Culex pipiens, Cx. quinquefasciatus, and Cx. molestus are important pest species and vectors of human and animal diseases throughout the world's tropical, temperate, and Holarctic regions. Diseases transmitted by member of the complex include: St. Louis encephali...

  3. HISTOCHEMICAL EFFECTS OF SOME BIOLOGICAL AGENTS ON CULEX PIPIENS LARVAE.

    PubMed

    El Sobky, Mona M; Ismail, Howaida I H; Assar, Abada A

    2016-04-01

    The histochemical effects of the lethal concentration that kills 50% of larvae (LC50) of three biological agents, abamectin, Bacillus thuringiensis and spinosad on the carbohydrates (polysaccharides), proteins, nucleic acids and lipids content of the midgut and fat bodies of Culex pipiens 2nd instar larvae were studied. The results showed that the three tested compounds reduced the carbohydrates (polysaccharides), proteins, RNA synthesis and lipids content after 72 hours of treatment where abamectin was the most effective followed by Bacillus thuringiensis then spinosad. PMID:27363043

  4. Culex pipiens, an Experimental Efficient Vector of West Nile and Rift Valley Fever Viruses in the Maghreb Region

    PubMed Central

    Amraoui, Fadila; Krida, Ghazi; Bouattour, Ali; Rhim, Adel; Daaboub, Jabeur; Harrat, Zoubir; Boubidi, Said-Chawki; Tijane, Mhamed; Sarih, Mhammed; Failloux, Anna-Bella

    2012-01-01

    West Nile fever (WNF) and Rift Valley fever (RVF) are emerging diseases causing epidemics outside their natural range of distribution. West Nile virus (WNV) circulates widely and harmlessly in the old world among birds as amplifying hosts, and horses and humans as accidental dead-end hosts. Rift Valley fever virus (RVFV) re-emerges periodically in Africa causing massive outbreaks. In the Maghreb, eco-climatic and entomologic conditions are favourable for WNV and RVFV emergence. Both viruses are transmitted by mosquitoes belonging to the Culex pipiens complex. We evaluated the ability of different populations of Cx. pipiens from North Africa to transmit WNV and the avirulent RVFV Clone 13 strain. Mosquitoes collected in Algeria, Morocco, and Tunisia during the summer 2010 were experimentally infected with WNV and RVFV Clone 13 strain at titers of 107.8 and 108.5 plaque forming units/mL, respectively. Disseminated infection and transmission rates were estimated 14–21 days following the exposure to the infectious blood-meal. We show that 14 days after exposure to WNV, all mosquito st developed a high disseminated infection and were able to excrete infectious saliva. However, only 69.2% of mosquito strains developed a disseminated infection with RVFV Clone 13 strain, and among them, 77.8% were able to deliver virus through saliva. Thus, Cx. pipiens from the Maghreb are efficient experimental vectors to transmit WNV and to a lesser extent, RVFV Clone 13 strain. The epidemiologic importance of our findings should be considered in the light of other parameters related to mosquito ecology and biology. PMID:22693557

  5. Modeling the distribution of the West Nile and Rift Valley Fever vector Culex pipiens in arid and semi-arid regions of the Middle East and North Africa

    PubMed Central

    2014-01-01

    Background The Middle East North Africa (MENA) region is under continuous threat of the re-emergence of West Nile virus (WNV) and Rift Valley Fever virus (RVF), two pathogens transmitted by the vector species Culex pipiens. Predicting areas at high risk for disease transmission requires an accurate model of vector distribution, however, most Cx. pipiens distribution modeling has been confined to temperate, forested habitats. Modeling species distributions across a heterogeneous landscape structure requires a flexible modeling method to capture variation in mosquito response to predictors as well as occurrence data points taken from a sufficient range of habitat types. Methods We used presence-only data from Egypt and Lebanon to model the population distribution of Cx. pipiens across a portion of the MENA that also encompasses Jordan, Syria, and Israel. Models were created with a set of environmental predictors including bioclimatic data, human population density, hydrological data, and vegetation indices, and built using maximum entropy (Maxent) and boosted regression tree (BRT) methods. Models were created with and without the inclusion of human population density. Results Predictions of Maxent and BRT models were strongly correlated in habitats with high probability of occurrence (Pearson’s r = 0.774, r = 0.734), and more moderately correlated when predicting into regions that exceeded the range of the training data (r = 0.666,r = 0.558). All models agreed in predicting high probability of occupancy around major urban areas, along the banks of the Nile, the valleys of Israel, Lebanon, and Jordan, and southwestern Saudi Arabia. The most powerful predictors of Cx. pipiens habitat were human population density (60.6% Maxent models, 34.9% BRT models) and the seasonality of the enhanced vegetation index (EVI) (44.7% Maxent, 16.3% BRT). Maxent models tended to be dominated by a single predictor. Areas of high probability corresponded with sites of

  6. Mosquito larvicidal activity of aromatic medicinal plant oils against Aedes aegypti and Culex pipiens pallens.

    PubMed

    Lee, Hoi-Seon

    2006-06-01

    Larvicidal activity of essential oils derived from 11 aromatic medicinal plants against early 4th-stage larvae of Aedes aegypti and Culex pipiens pallens was tested in the laboratory. At 100 ppm, the essential oils of all plants caused 100% mortality against Ae. aegypti and Cx. pipiens pallens. At 25 ppm, the essential oils of Citrus bergamia, Cuminum myrrha, and Pimenta racemosa caused 100% mortality against larvae of Ae. aegypti and Cx. pipiens pallens. The oil of C. begamia caused 32.5% and 24.5% mortality against Ae. aegypti and Cx. pipiens pallens at 12.5 ppm, but 24.2% and 0% mortality against Ae. aegypti and Cx. pipiens pallens at 6.25 ppm, respectively. The oil of P. racemosa caused 52.3% and 38.5% mortality against Ae. aegypti and Cx. pipiens pallens at 12.5 ppm, but 32.2% and 0% mortality against Ae. aegypti and Cx. pipiens pallens at 6.25 ppm, respectively. The larvicidal activity of oils of C. bergamia, C. myrrha, and P. racemosa was significantly reduced when used at 6.25 ppm. These plants warrant further studies as possible agents for mosquito control. PMID:17019775

  7. Columbia spotted frog (Rana luteiventris) in southeastern Oregon: A survey of historical localities, 2009

    USGS Publications Warehouse

    Pearl, Chistopher A.; Galvan, Stephanie K.; Adams, Michael J.; McCreary, Brome

    2010-01-01

    The Columbia spotted frog (Rana luteiventris) occupies a large range in western North America and is comprised of at least three genetic units. Concern exists regarding the status of the Great Basin populations in Oregon, Idaho, and Nevada. We surveyed target and nearby alternate sites on public lands in southeastern Oregon where there was evidence that Columbia spotted frogs were historically present. We found the species at 59.5 percent (25 of 42) of target or nearby alternate sites. They were in 15 of 23 permanent streams and 8 of 13 intermittent streams. Our surveys do not provide evidence of widespread population losses in our sites. Interpretation of status of Columbia spotted frogs in this study is limited by a lack of precision in some of the historical locations and by our inability to determine if locations where only adults were indicated in the historical record once had breeding populations. Our results support the need for continued investigation of these populations.

  8. Mosquito surveillance for West Nile virus in Connecticut, 2000: isolation from Culex pipiens, Cx. restuans, Cx. salinarius, and Culiseta melanura.

    PubMed Central

    Andreadis, T. G.; Anderson, J. F.; Vossbrinck, C. R.

    2001-01-01

    Fourteen isolations of West Nile (WN) virus were obtained from four mosquito species (Culex pipiens [5], Cx. restuans [4], Cx. salinarius [2], and Culiseta melanura [3]) in statewide surveillance conducted from June through October 2000. Most isolates were obtained from mosquitoes collected in densely populated residential locales in Fairfield and New Haven counties, where the highest rates of dead crow sightings were reported and where WN virus was detected in 1999. Minimum field infection rates per 1,000 mosquitoes ranged from 0.5 to 1.8 (county based) and from 1.3 to 76.9 (site specific). Cx. restuans appears to be important in initiating WN virus transmission among birds in early summer; Cx. pipiens appears to play a greater role in amplifying virus later in the season. Cs. melanura could be important in the circulation of WN virus among birds in sylvan environments; Cx. salinarius is a suspected vector of WN virus to humans and horses. PMID:11585530

  9. Introduction to the symposium Global Perspective on the Culex pipiens Complex in the 21st century: The Interrelationship of Culex pipiens, quinquefasciatus, molestus and others.

    PubMed

    Linthicum, Kenneth J

    2012-12-01

    Mosquitoes in the Culex pipiens Complex, including Culex pipiens, Cx. quinquefasciatus, and Cx. molestus, are important pest species and vectors of human and animal diseases throughout the world's tropical, temperate, and Holarctic regions. Diseases transmitted by member of the Pipiens Complex include: St. Louis encephalitis and West Nile virus in North America, West Nile virus on several other continents, Rift Valley fever in Africa, lymphatic filariasis caused by Wuchereria bancrofti in the tropics, and Dirofilaria immitis globally. Here and in the following 14 papers, 3 abstracts and a summary paper are the proceedings of a symposium that gathered many of the world's experts on the Pipiens Complex to explore the current state of knowledge of the taxa. Information presented at the symposium will improve our knowledge of important members of the complex and enhance our ability to conduct efficient surveillance and efficacious control strategies. A background on previous discussions on the Pipiens Complex, and a brief description of current symposium contributors and their topics are discussed. PMID:23401940

  10. The wood frog (Rana sylvatica): a technical conservation assessment

    USGS Publications Warehouse

    Muths, E.; Rittmann, S.; Irwin, J.; Keinath, D.; Scherer, R.

    2005-01-01

    Overall, the wood frog (Rana sylvatica) is ranked G5, secure through most of its range (NatureServe Explorer 2002). However, it is more vulnerable in some states within the USDA Forest Service Region 2: S3 (vulnerable) in Colorado, S2 (imperiled) in Wyoming, and S1 (critically imperiled in South Dakota (NatureServe Explorer 2002); there are no records for wood frogs in Kansas or Nebraska. Primary threats to wood frog populations are habitat fragmentation (loss of area, edge effects, and isolation) and habitat loss due to anthropogenic causes (e.g., wetland draining, grazing) and natural changes as habitat succession occurs. Wood frogs are most conspicuous at breeding sites early in the spring, when snow and ice are often still present at pond margins. They tolerate frezzing and hibernate terrestrially in shallow depressions, under leaf litter, grasses, logs, or rocks (Bagdonas 1968, Bellis 1961a); there are no reports of aquatic hibernation for this species (Licht 1991, Pinder et al. 1992). Wood frogs require semi-permanent and temporary pools of natural origin and adjacent wet meadows, and landscape alterations that shorten the hydroperiod of ponds can result in catastrophic tadpole mortality. Plant communities utilized by wood frogs in the Rocky Mountains are hydric to mesic and include sedge and grass meadows, willow hummocks, aspen groves, lodgepole pine forests, and woodlands with leaf litter and/or herbaceous understory (Maslin 1947, Bellis 1961a, Roberts and Lewin 1979, Haynes and Aird 1981). Wood frogs are likely to disperse into surrounding marsh and woodlands soon after oviposition (Heatwole 1961, Haynes and Aird 1981). In the arly fall, wood frogs begin to seek hibernacula at or just below the ground surface, generally in upland forest habitat (Regosin et al. 2003). Licht (1991) demonstrated shelter-seeking behavior at 1.5 [degrees] C. Once they have concealed themselves for hibernation, wood frogs are very difficult to detecta?|

  11. Acute toxicity of furazolidone on Artemia salina, Daphnia magna, and Culex pipiens molestus larvae

    SciTech Connect

    Macri, A.; Stazi, A.V.; Dojmi di Delupis, G.

    1988-10-01

    As a result of evidence of the ecotoxicity of nitrofurans, the acute toxicity of furazolidone was tested in vivo on two aquatic organisms, Artemia salina and Daphnia magna, which are both crustaceans. Toxicity studies were also performed on larvae of Culex pipiens molestus. Results indicated a significant toxicity of the compound on Culex pipiens and Daphnia magna, while Artemia salina proved to be the least sensitive.

  12. Density dependent growth in adult brown frogs Rana arvalis and Rana temporaria - A field experiment

    NASA Astrophysics Data System (ADS)

    Loman, Jon; Lardner, Björn

    2009-11-01

    In species with complex life cycles, density regulation can operate on any of the stages. In frogs there are almost no studies of density effects on the performance of adult frogs in the terrestrial habitat. We therefore studied the effect of summer density on the growth rate of adult frogs during four years. Four 30 by 30 m plots in a moist meadow were used. In early summer, when settled after post-breeding migration, frogs ( Rana arvalis and Rana temporaria that have a very similar ecology and potentially compete) were enclosed by erecting a fence around the plots. Frogs were captured, measured, marked and partly relocated to create two high density and two low density plots. In early autumn the frogs were again captured and their individual summer growth determined. Growth effects were evaluated in relation to two density measures: density by design (high/low manipulation), and actual (numerical) density. R. arvalis in plots with low density by design grew faster than those in high density plots. No such effect was found for R. temporaria. For none of the species was growth related to actual summer density, determined by the Lincoln index and including the density manipulation. The result suggests that R. arvalis initially settled according to an ideal free distribution and that density had a regulatory effect (mediated through growth). The fact that there were no density effects on R. temporaria (and a significant difference in its response to that of R. arvalis) suggests it is a superior competitor to R. arvalis during the terrestrial phase. There were no density effects on frog condition index, suggesting that the growth rate modifications may actually be an adaptive trait of R. arvalis. The study demonstrates that density regulation may be dependent on resources in frogs' summer habitat.

  13. Exposure to West Nile Virus Increases Bacterial Diversity and Immune Gene Expression in Culex pipiens

    PubMed Central

    Zink, Steven D.; Van Slyke, Greta A.; Palumbo, Michael J.; Kramer, Laura D.; Ciota, Alexander T.

    2015-01-01

    Complex interactions between microbial residents of mosquitoes and arboviruses are likely to influence many aspects of vectorial capacity and could potentially have profound effects on patterns of arbovirus transmission. Such interactions have not been well studied for West Nile virus (WNV; Flaviviridae, Flavivirus) and Culex spp. mosquitoes. We utilized next-generation sequencing of 16S ribosomal RNA bacterial genes derived from Culex pipiens Linnaeus following WNV exposure and/or infection and compared bacterial populations and broad immune responses to unexposed mosquitoes. Our results demonstrate that WNV infection increases the diversity of bacterial populations and is associated with up-regulation of classical invertebrate immune pathways including RNA interference (RNAi), Toll, and Jak-STAT (Janus kinase-Signal Transducer and Activator of Transcription). In addition, WNV exposure alone, without the establishment of infection, results in similar alterations to microbial and immune signatures, although to a lesser extent. Multiple bacterial genera were found in greater abundance in WNV-exposed and/or infected mosquitoes, yet the most consistent and notable was the genus Serratia. PMID:26516902

  14. Attractant and stimulant factors for oviposition of Culex pipiens fatigans in natural breeding-sites*

    PubMed Central

    Ikeshoji, Toshiaki

    1966-01-01

    The breeding of mosquito larvae in the field is determined by the ovipositing behaviour of the gravid females. Investigation of the chemical factors that induce oviposition is therefore important for understanding mosquito ecology. These substances may also prove to be useful in assessing and controlling mosquito populations. The author has demonstrated two chemical factors, an ovipositing attractant and an ovipositing stimulant, in surface-water. The ovipositing attractant, extracted from surface-water by distillation and extraction with diethyl ether, was found to be quite effective when used to recapture known numbers of gravid mosquitos released in a large calf-shed. The presence of the stimulant factor was established by forcing gravid females to touch the testing water with tarsi and proboscis. After such contact, they began oviposition three times more rapidly on surface-water than on tap-water. The importance of these factors was demonstrated in the larval populations of Culex pipiens fatigans in pit-latrines in Rangoon, Burma. PMID:5298039

  15. Tracking the evolution of insecticide resistance in the mosquito Culex pipiens.

    PubMed

    Lenormand, T; Bourguet, D; Guillemaud, T; Raymond, M

    1999-08-26

    The evolution of pesticide resistance provides some of the most striking examples of darwinian evolution occurring over a human life span. Identification of resistance alleles opens an outstanding framework in which to study the evolution of adaptive mutations from the beginning of pesticide application, the evolution of interactions between alleles (dominance) or between loci (epistasis). Here we show that resistance alleles can also be used as markers to dissect population processes at a microevolutionary scale. We have focused on the antagonistic roles of selection and migration involved in the dynamics of local adaptation with reference to allelic frequencies at two resistance loci in the mosquito Culex pipiens. We find that their frequencies follow an annual cycle of large amplitude (25%), and we precisely unravel the seasonal variation of migration and selection underlying this cycle. Our results provide a firm basis on which to devise an insecticide treatment strategy that will better control the evolution of resistance genes and the growth of mosquito populations. PMID:10476962

  16. Pathophysiology in mountain yellow-legged frogs (Rana muscosa) during a chytridiomycosis outbreak.

    PubMed

    Voyles, Jamie; Vredenburg, Vance T; Tunstall, Tate S; Parker, John M; Briggs, Cheryl J; Rosenblum, Erica Bree

    2012-01-01

    The disease chytridiomycosis is responsible for declines and extirpations of amphibians worldwide. Chytridiomycosis is caused by a fungal pathogen (Batrachochytrium dendrobatidis) that infects amphibian skin. Although we have a basic understanding of the pathophysiology from laboratory experiments, many mechanistic details remain unresolved and it is unknown if disease development is similar in wild amphibian populations. To gain a better understanding of chytridiomycosis pathophysiology in wild amphibian populations, we collected blood biochemistry measurements during an outbreak in mountain yellow-legged frogs (Rana muscosa) in the Sierra Nevada Mountains of California. We found that pathogen load is associated with disruptions in fluid and electrolyte balance, yet is not associated with fluctuations acid-base balance. These findings enhance our knowledge of the pathophysiology of this disease and indicate that disease development is consistent across multiple species and in both laboratory and natural conditions. We recommend integrating an understanding of chytridiomycosis pathophysiology with mitigation practices to improve amphibian conservation. PMID:22558145

  17. Artificial Selection for Different Host Preferences in Culex pipiens pallens (Diptera: Culicidae) Mosquitoes.

    PubMed

    Yu, Jing; Li, Chun-Xiao; Dong, Yan-De; Xue, Rui-De; Zhao, Tong-Yan

    2015-09-01

    Most mosquito species display host preferences that are a crucial determinant of the transmission rate of mosquito-borne pathogens. Although a transgenic approach, based on driving genes for zoophily into vector populations, has been advocated as a malaria control strategy by the World Health Organization since 1982, the genes involved in mosquito host choice remain poorly understood. Culex pipiens pallens Coquillet mosquitoes were artificially selected for two different host preferences in a specially designed experimental enclosure. Of 3,035 mosquitoes obtained from larvae and pupae collected from the wild (the F0 generation), 27% preferentially fed on pigeons and 16% fed on mice. Following artificial selection for these host preferences over successive generations, the percentage of mosquitoes that preferred to feed on pigeons or mice gradually increased, eventually stabilizing at ∼55 and 34%, respectively, after the sixth generation. Intergenerational differences in host preferences were significant (P < 0.001). Furthermore, differences in host preferences between mosquitoes selected to prefer pigeons and those selected to prefer mice were both significant and consistent over almost six generations. PMID:26336219

  18. Fitness consequences of infection by Batrachochytrium dendrobatidis in northern leopard frogs (Lithobates pipiens).

    PubMed

    Chatfield, Matthew W H; Brannelly, Laura A; Robak, Matthew J; Freeborn, Layla; Lailvaux, Simon P; Richards-Zawacki, Corinne L

    2013-03-01

    The amphibian chytrid fungus, Batrachochytrium dendrobatidis (Bd), has been linked to amphibian declines and extinctions worldwide. The pathogen has been found on amphibians throughout eastern North America, but has not been associated with mass die-offs in this region. In this study, we conducted laboratory experiments on the effects of Bd infection in a putative carrier species, Lithobates pipiens, using two estimators of fitness: jumping performance and testes morphology. Over the 8-week study period, peak acceleration during jumping was not significantly different between infected and uninfected animals. Peak velocity, however, was significantly lower for infected animals after 8 weeks. Two measures of sperm production, germinal epithelium depth, and maximum spermatic cyst diameter, showed no difference between infected and uninfected animals. The width, but not length, of testes of infected animals was significantly greater than in uninfected animals. This study is the first to show effects on whole-organism performance of Bd infection in post-metamorphic amphibians, and may have important long-term, evolutionary implications for amphibian populations co-existing with Bd infection. PMID:23604643

  19. Quantification of intrahost bottlenecks of West Nile virus in Culex pipiens mosquitoes using an artificial mutant swarm

    PubMed Central

    Ciota, Alexander T.; Ehrbar, Dylan J.; Van Slyke, Greta A.; Payne, Anne F.; Willsey, Graham G.; Viscio, Rachael E.; Kramer, Laura D.

    2012-01-01

    Mosquito-borne viruses are predominantly RNA viruses which exist within hosts as diverse mutant swarms. Defining the way in which stochastic forces within mosquito vectors shape these swarms is critical to advancing our understanding of the evolutionary and adaptive potential of these pathogens. There are multiple barriers within a mosquito which a viral swarm must traverse in order to ultimately be transmitted. Here, using artificial mutant swarms composed of neutral variants of West Nile virus (WNV), we tracked changes to swarm breadth over time and space in Culex pipiens mosquitoes. Our results demonstrate that all variants have the potential to survive intrahost bottlenecks, yet mean swarm breadth decreases during both midgut infection and transmission when starting populations contain higher levels of minority variants. In addition, WNV swarms are subject to temporal sweeps which act to significantly decrease intrahost diversity over time. Taken together, these data demonstrate the profound effects that stochastic forces can have in shaping arboviral mutant swarms. PMID:22326536

  20. A Pictorial Key for Culex pipiens Complex (Diptera: Culicidae) In Iran

    PubMed Central

    Dehghan, Hossein; Sadraei, Javid; Moosa-Kazemi, Seyed Hassan; Abolghasemi, Esmail; Solimani, Hassan; Jaffari Nodoshan, Ahmad; Najafi, Mohammad Hassan

    2016-01-01

    Background: The aim of this study was to design pictorial key and taxonomic literature of Culex pipiens complex in Iran. Methods: Larvae were collected using standard dipping methods in 13 randomly selected areas of Bushehr, Hamedan, Kerman, Khorasan-e-Razavi, Khuzistan, Mazandaran, Tehran, Sistan and Baluchistan and Yazd Provinces from April 2009 to October 2010. The data were analyzed using SPSS Ver. 11.5. Results: Culex pipiens larvae were identified based on the Seta 1 of the abdominal segments III–IV in north and central parts of Iran. This diagnostic character had some variation among the Cx. quinquefasciatus collected from south of the country. The identification value of intersection of costa, subcosta and bifurcation of R2+3 of female veins, was calculated as 90–100 % for Cx. pipiens. This diagnostic character was varied among the Cx. quinquefasciatus specimens. The male genitalia found as the main characters to distinguish of Cx. quinquefasciatus from Cx. pipiens. Conclusion: It is necessary more studies on the behavior and genetic variations of Cx. pipiens complex in Iran. PMID:27308288

  1. Spatial Heterogeneity of Rana boylii Habitat: Quantification and Ecological Meaningfulness

    NASA Astrophysics Data System (ADS)

    Yarnell, S. M.

    2005-05-01

    Analysis of the heterogeneity of stream habitat and how biological communities respond to that complexity are fundamental components of ecosystem analysis that are often inadequately addressed in watershed assessments and restoration practices. Many aquatic species, such as the Foothill Yellow-legged Frog (Rana boylii), known to associate with certain physical habitats at various times throughout their lifecycle may require some degree of habitat complexity at a larger reach scale for a population to persist. Recent research in the field of landscape ecology has expanded the use of spatial heterogeneity indices to other fields of ecology as an objective method to quantify variability in habitat. Provided that indices are used in an appropriate context and are shown to be ecologically meaningful, they provide a potentially useful tool for quantifying the variability in riverine habitat for aquatic species such as R. boylii. This study evaluated whether stream reaches with a high heterogeneity of geomorphic features, as measured by several key spatial heterogeneity indices, correlated with a greater relative abundance of R. boylii. R. boylii habitat associations were quantified throughout a single season to obtain further insight into the local hydraulic and geomorphic conditions preferred by each lifestage. The two best predictors of habitat associations by lifestage were velocity and substrate size, two key characteristics of geomorphic units such as riffles and pools. The heterogeneity of geomorphic units was then quantified and measured at the reach scale using a variety of spatial indices. Indices of spatial composition, such as Shannon's Diversity Index, were found to correlate well with frog abundance, while indices of spatial configuration, such as Contagion, were not significant. These findings indicate R. boylii may select stream reaches with increased geomorphic complexity that potentially provide habitats suitable to each lifestage with multiple functions

  2. Culex pipiens quinquefasciatus: a potential vector to transmit Zika virus.

    PubMed

    Guo, Xiao-Xia; Li, Chun-Xiao; Deng, Yong-Qiang; Xing, Dan; Liu, Qin-Mei; Wu, Qun; Sun, Ai-Juan; Dong, Yan-de; Cao, Wu-Chun; Qin, Cheng-Feng; Zhao, Tong-Yan

    2016-01-01

    Zika virus (ZIKV) has become a threat to global health since the outbreak in Brazil in 2015. Although ZIKV is generally considered an Aedes-transmitted pathogen, new evidence has shown that parts of the virus closely resemble Culex-transmitted viruses. Therefore, it is important to evaluate the competence of Culex species for ZIKV to understand their potential as vectors. In this study, female Culex pipiens quinquefasciatus were orally exposed to ZIKV. Mosquito midguts, salivary glands and ovaries were tested for ZIKV to measure infection and dissemination at 2, 4, 6, 8, 12, 16 and 18 days post exposure (pe). In addition, saliva was collected from mosquitoes after infection and infant mice were bitten by infected mosquitoes to measure the transmission ability of Cx. p. quinquefasciatus. The results showed that the peak time of virus appearance in the salivary glands was day 8 pe, with 90% infection rate and an estimated virus titer of 3.92±0.49 lg RNA copies/mL. Eight of the nine infant mice had positive brains after being bitten by infected mosquitoes, which meant that Cx. p. quinquefasciatus could be infected with and transmit ZIKV following oral infection. These laboratory results clearly demonstrate the potential role of Cx. p. quinquefasciatus as a vector of ZIKV in China. Because there are quite different vector management strategies required to control Aedes (Stegomyia) species and Cx. p. quinquefasciatus, an integrated approach may be required should a Zika epidemic occur. PMID:27599470

  3. Optical Spectra of Melanin Films Extracted from Rana esculenta L.

    NASA Astrophysics Data System (ADS)

    Perna, G.; Gallone, A.; Capozzi, V.; Biagi, P. F.; Fratello, A.; Guida, G.; Zanna, P.; Argenzio, E.; Cicero, R.

    2005-01-01

    The melanin pigment extracted from the liver of Rana esculenta L. has been deposited as thin film on quartz substrate. The Raman spectra, as well as optical absorption and photoluminescence measurements have been investigated. The results show that the melanin can be described as a network of clusters having different size. The larger size clusters determine the absorption edge of the film and the smaller size ones are mainly involved in the radiative emission process.

  4. Comparative toxicity of chlorpyrifos, diazinon, malathion and their oxon derivatives to larval Rana boylii

    USGS Publications Warehouse

    Sparling, D.W.; Fellers, G.

    2007-01-01

    Organophosphorus pesticides (OPs) are ubiquitous in the environment and are highly toxic to amphibians. They deactivate cholinesterase, resulting in neurological dysfunction. Most chemicals in this group require oxidative desulfuration to achieve their greatest cholinesterase-inhibiting potencies. Oxon derivatives are formed within liver cells but also by bacterial decay of parental pesticides. This study examines the toxicity of chlorpyrifos, malathion and diazinon and their oxons on the foothill yellow-legged frog (Rana boylii). R. boylii is exposed to agricultural pesticides in the California Central Valley. Median lethal concentrations of the parental forms during a 96 h exposure were 3.00 mg/L (24 h) for chlorpyrifos, 2.14 mg/L for malathion and 7.49 mg/L for diazinon. Corresponding oxons were 10 to 100 times more toxic than their parental forms. We conclude that environmental concentrations of these pesticides can be harmful to R. boylii populations. ?? 2006 Elsevier Ltd. All rights reserved.

  5. Toxicity of some plant extracts against vector of lymphatic filariasis, Culex pipiens.

    PubMed

    Hasaballah, Ahmed I

    2015-04-01

    Many insecticides are generally used as larvicides to control Culex pipiens, vector of lymphatic filariasis. This study was undertaken to evaluate the larvicidal activity of some potential larvicidal plants extracts against C. pipiens larvae. The toxic effects of both ethanolic and petroleum ether plant extracts were evaluated under laboratory conditions against 3rd instar larvae of C. pipiens. Forty ethanolic and petroleum ether extracts of 10 plants namely Echinochloa stagninum, Phragmites australis, Eichhornia crassipes, Rhizophora mucronata, Cichorium intybus, Ocimum basilicum, Origanum majorana, Azadirachta indica, Rosmarinus officinalis and Nigella sativa. On the basis of LC50, the toxic effect of the plant extracts tested varied depending on the plant species, part, solvent used in extraction and the extract concentrations. The petroleum ether extraction was more effective against mosquito as compared with ethanolic extraction. The most effective plant extract was A. indica followed by Ph. australis, N. sativa, C. intybus, R. officinalis, O. basilicum, O. majorana, E. stagninum, Rh. Mucronata and E. crassipes. PMID:26012233

  6. Comparative fine structure of eggs of autogenous and anautogenous Culex pipiens (Diptera: Culicidae).

    PubMed

    Soliman, Belal A; Tewfick, Maha K; Wassim, Nahla M

    2014-12-01

    Culex (Cx.) pipiens is the potential vector human filariasis in Egypt. However, autogenous Cx. pipiens may be less efficient vector of Wuchereria (W.) bancrofti in endemic areas of Egypt compared to anautogenous counterparts. In this study, an attempt was made to differentiate eggs of autogenous and anautogenous Cx. pipiens using scanning electron micrographs. The results indicated that eggs of both species appear to be similar to great extent in surface morphology. Eggs of both forms are black and elongate-oval. Width is greatest at the anterior end. The posterior end is pointed. The micropylar disc is apparent with distinct edge. Exochorionic bridges are angular. Size of both eggs represented by length and width are comparable. In both eggs, length is greater than width. However, eggs of both forms can be distinguished from each other by the exochorionic bridges being longer and thinner in the autogenous eggs than in the anautogenous eggs. PMID:25643517

  7. “BIRD BITING” MOSQUITOES AND HUMAN DISEASE: A REVIEW OF THE ROLE OF CULEX PIPIENS COMPLEX MOSQUITOES IN EPIDEMIOLOGY

    PubMed Central

    Farajollahi, Ary; Fonseca, Dina M.; Kramer, Laura D.; Kilpatrick, A. Marm

    2011-01-01

    The transmission of vector-borne pathogens is greatly influenced by the ecology of their vector, which is in turn shaped by genetic ancestry, the environment, and the hosts it feeds on. One group of vectors, the mosquitoes in the Culex pipiens complex, play key roles in the transmission of a range of pathogens including several viruses such as West Nile and St. Louis encephalitis viruses, avian malaria (Plasmodium spp.), and filarial worms. The Cx. pipiens complex includes Cx. pipiens pipiens with two forms, pipiens and molestus, Cx. pipiens pallens, Cx. quinquefasciatus, Cx. australicus, and Cx. globocoxitus. While several members of the complex have limited geographic distributions, Cx. pipiens pipiens and Cx. quinquefasciatus are found in all known urban and sub-urban temperate and tropical regions, respectively, across the world, where they are often principal disease vectors. In addition, hybrids are common in areas of overlap. Although gaps in our knowledge still remain, the advent of genetic tools has greatly enhanced our understanding of the history of speciation, domestication, dispersal, and hybridization. We review the taxonomy, genetics, evolution, behavior, and ecology, of members of the Cx. pipiens complex and their role in the transmission of medically important pathogens. The adaptation of Cx. pipiens complex mosquitoes to human-altered environments led to their global distribution through dispersal via humans and, combined with their mixed feeding patterns on birds and mammals (including humans), increased the transmission of several avian pathogens to humans. We highlight several unanswered questions that will increase our ability to control diseases transmitted by these mosquitoes. PMID:21875691

  8. Host selection by Culex pipiens mosquitoes and West Nile virus amplification.

    PubMed

    Hamer, Gabriel L; Kitron, Uriel D; Goldberg, Tony L; Brawn, Jeffrey D; Loss, Scott R; Ruiz, Marilyn O; Hayes, Daniel B; Walker, Edward D

    2009-02-01

    Recent field studies have suggested that the dynamics of West Nile virus (WNV) transmission are influenced strongly by a few key super spreader bird species that function both as primary blood hosts of the vector mosquitoes (in particular Culex pipiens) and as reservoir-competent virus hosts. It has been hypothesized that human cases result from a shift in mosquito feeding from these key bird species to humans after abundance of the key birds species decreases. To test this paradigm, we performed a mosquito blood meal analysis integrating host-feeding patterns of Cx. pipiens, the principal vector of WNV in the eastern United States north of the latitude 36 degrees N and other mosquito species with robust measures of host availability, to determine host selection in a WNV-endemic area of suburban Chicago, Illinois, during 2005-2007. Results showed that Cx. pipiens fed predominantly (83%) on birds with a high diversity of species used as hosts (25 species). American robins (Turdus migratorius) were marginally overused and several species were underused on the basis of relative abundance measures, including the common grackle (Quiscalus quiscula), house sparrow (Passer domesticus), and European starling (Sturnus vulgaris). Culex pipiens also fed substantially on mammals (19%; 7 species with humans representing 16%). West Nile virus transmission intensified in July of both years at times when American robins were heavily fed upon, and then decreased when robin abundance decreased, after which other birds species were selected as hosts. There was no shift in feeding from birds to mammals coincident with emergence of human cases. Rather, bird feeding predominated when the onset of the human cases occurred. Measures of host abundance and competence and Cx. pipiens feeding preference were combined to estimate the amplification fractions of the different bird species. Predictions were that approximately 66% of WNV-infectious Cx. pipiens became infected from feeding on just

  9. Culex pipiens Development Is Greatly Influenced by Native Bacteria and Exogenous Yeast.

    PubMed

    Díaz-Nieto, Leonardo M; D Alessio, Cecilia; Perotti, M Alejandra; Berón, Corina M

    2016-01-01

    Culex pipiens is the most cosmopolitan mosquito of the Pipiens Assemblage. By studying the nature of interactions between this species and microorganisms common to its breeding environment we can unravel important pitfalls encountered during development. We tested the survival rate of larval stages, pupae and adults of a Cx. pipiens colony exposed to a variety of microorganisms in laboratory conditions and assessed the transmission to offspring (F1) by those organisms that secured development up to adulthood. Three complementary experiments were designed to: 1) explore the nutritional value of yeasts and other microorganisms during Cx. pipiens development; 2) elucidate the transstadial transmission of yeast to the host offspring; and 3) to examine the relevance of all these microorganisms in female choice for oviposition-substratum. The yeast Saccharomyces cerevisiae proved to be the most nutritional diet, but despite showing the highest survival rates, vertical transmission to F1 was never confirmed. In addition, during the oviposition trials, none of the gravid females was attracted to the yeast substratum. Notably, the two native bacterial strains, Klebsiella sp. and Aeromonas sp., were the preferred oviposition media, the same two bacteria that managed to feed neonates until molting into 2nd instar larvae. Our results not only suggest that Klebsiella sp. or Aeromonas sp. serve as attractants for oviposition habitat selection, but also nurture the most fragile instar, L1, to assure molting into a more resilient stage, L2, while yeast proves to be the most supportive diet for completing development. These experiments unearthed survival traits that might be considered in the future development of strategies of Cx. pipiens control. These studies can be extended to other members of the Pipiens Assemblage. PMID:27055276

  10. Culex pipiens Development Is Greatly Influenced by Native Bacteria and Exogenous Yeast

    PubMed Central

    Díaz-Nieto, Leonardo M.; D´Alessio, Cecilia

    2016-01-01

    Culex pipiens is the most cosmopolitan mosquito of the Pipiens Assemblage. By studying the nature of interactions between this species and microorganisms common to its breeding environment we can unravel important pitfalls encountered during development. We tested the survival rate of larval stages, pupae and adults of a Cx. pipiens colony exposed to a variety of microorganisms in laboratory conditions and assessed the transmission to offspring (F1) by those organisms that secured development up to adulthood. Three complementary experiments were designed to: 1) explore the nutritional value of yeasts and other microorganisms during Cx. pipiens development; 2) elucidate the transstadial transmission of yeast to the host offspring; and 3) to examine the relevance of all these microorganisms in female choice for oviposition-substratum. The yeast Saccharomyces cerevisiae proved to be the most nutritional diet, but despite showing the highest survival rates, vertical transmission to F1 was never confirmed. In addition, during the oviposition trials, none of the gravid females was attracted to the yeast substratum. Notably, the two native bacterial strains, Klebsiella sp. and Aeromonas sp., were the preferred oviposition media, the same two bacteria that managed to feed neonates until molting into 2nd instar larvae. Our results not only suggest that Klebsiella sp. or Aeromonas sp. serve as attractants for oviposition habitat selection, but also nurture the most fragile instar, L1, to assure molting into a more resilient stage, L2, while yeast proves to be the most supportive diet for completing development. These experiments unearthed survival traits that might be considered in the future development of strategies of Cx. pipiens control. These studies can be extended to other members of the Pipiens Assemblage. PMID:27055276

  11. Conservation genetics of evolutionary lineages of the endangered mountain yellow-legged frog, Rana muscosa (Amphibia: Ranidae), in southern California

    USGS Publications Warehouse

    Schoville, Sean D.; Tustall, Tate S.; Vredenburg, Vance T.; Backlin, Adam R.; Gallegos, Elizabeth; Wood, Dustin A.; Fisher, Robert N.

    2011-01-01

    Severe population declines led to the listing of southern California Rana muscosa (Ranidae) as endangered in 2002. Nine small populations inhabit watersheds in three isolated mountain ranges, the San Gabriel, San Bernardino and San Jacinto. One population from the Dark Canyon tributary in the San Jacinto Mountains has been used to establish a captive breeding population at the San Diego Zoo Institute for Conservation Research. Because these populations may still be declining, it is critical to gather information on how genetic variation is structured in these populations and what historical inter-population connectivity existed between populations. Additionally, it is not clear whether these populations are rapidly losing genetic diversity due to population bottlenecks. Using mitochondrial and microsatellite data, we examine patterns of genetic variation in southern California and one of the last remaining populations of R. muscosa in the southern Sierra Nevada. We find low levels of genetic variation within each population and evidence of genetic bottlenecks. Additionally, substantial population structure is evident, suggesting a high degree of historical isolation within and between mountain ranges. Based on estimates from a multi-population isolation with migration analysis, these populations diversified during glacial episodes of the Pleistocene, with little gene flow during population divergence. Our data demonstrate that unique evolutionary lineages of R. muscosa occupy each mountain range in southern California and should be managed separately. The captive breeding program at Dark Canyon is promising, although mitigating the loss of neutral genetic diversity relative to the natural population might require additional breeding frogs.

  12. Highly polluted larval habitats of the Culex pipiens complex in central Sweden

    SciTech Connect

    Ishii, T.; Sohn, S.R.

    1987-06-01

    Larvae of the Culex pipiens complex (Cx. pipiens and Cx. torrentium) were abundant in two highly polluted pools receiving sewage sludge in Uppsala, Sweden (early August through late September 1985). The water was characterized by high BOD, and high ion concentration of Cu, Fe, Al and much suspended matter. Maximum larval number at the pool surface area was 26.1/ml. The ratio between species was studied and Cx. torrentium comprised ca. 20% at the peak of abundance. Some egg rafts showed no embryogeny.

  13. Population status and population genetics of northern leopard frogs in Arizona

    USGS Publications Warehouse

    Theimer, Tad C.; Drost, Charles A.; O'Donnell, Ryan P.; Mock, Karen E.

    2011-01-01

    Increasing isolation of populations by habitat fragmentation threatens the persistence of many species, both from stochastic loss of small isolated populations, and from inbreeding effects in populations that have become genetically isolated. In the southwestern United States, amphibian habitat is naturally patchy in occurrence because of the prevailing aridity of the region. Streams, rivers, and other wetlands are important both as habitat and as corridors that connect populations. However, populations of some species have become more fragmented and isolated by habitat degradation and loss. Northern leopard frogs (Rana pipiens) have experienced serious declines in the Southwest. We conducted an extensive survey across the known range of northern leopard frogs in Arizona to determine the current distribution and abundance of the species. From a range that once spanned much of the northern and central part of the State, northern leopard frogs have been reduced to three or four widely separated populations, near Lyman Lake in east-central Arizona, in the Stoneman Lake area south of Flagstaff, along Truxton Wash near Peach Springs, and a population of uncertain extent on Navajo Nation lands. The Lyman Lake and Truxton Wash populations are small and extremely isolated. The Stoneman Lake population, however, is an extensive metapopulation spread across several stream drainages, including numerous ponds, wetlands, and artificial tanks. This is the only population in Arizona that is increasing in extent and numbers, but there is concern about the apparent introduction of nonnative genetic stock from eastern North America into this area. We analyzed genetic diversity within and genetic divergence among populations of northern leopard frogs, across both extant and recently extirpated populations in Arizona. We also analyzed mitochondrial DNA to place these populations into a larger phylogenetic framework and to determine whether any populations contained genetic material

  14. ASSESSMENT OF THE RISK OF SOLAR ULTRAVIOLET RADIATION TO AMPHIBIANS. 1) DOSE-DEPENDENT INDUCTION OF HINDLIMB MALFORMATIONS IN THE NORTHERN LEOPARD FROG (RANA PIPIENS)

    EPA Science Inventory

    A number of environmental stressors have been hypothesized as responsible for seeming increases in limb malformations in several species of North American amphibians. The purpose of this study was to generate dose-response data suitable for assessing the potential role of solar u...

  15. Acid stress mediated adaptive divergence in ion channel function during embryogenesis in Rana arvalis

    PubMed Central

    Shu, Longfei; Laurila, Anssi; Räsänen, Katja

    2015-01-01

    Ion channels and pumps are responsible for ion flux in cells, and are key mechanisms mediating cellular function. Many environmental stressors, such as salinity and acidification, are known to severely disrupt ionic balance of organisms thereby challenging fitness of natural populations. Although ion channels can have several vital functions during early life-stages (e.g. embryogenesis), it is currently not known i) how developing embryos maintain proper intracellular conditions when exposed to environmental stress and ii) to what extent environmental stress can drive intra-specific divergence in ion channels. Here we studied the moor frog, Rana arvalis, from three divergent populations to investigate the role of different ion channels and pumps for embryonic survival under acid stress (pH 4 vs 7.5) and whether populations adapted to contrasting acidities differ in the relative role of different ion channel/pumps. We found that ion channels that mediate Ca2+ influx are essential for embryonic survival under acidic pH, and, intriguingly, that populations differ in calcium channel function. Our results suggest that adaptive divergence in embryonic acid stress tolerance of amphibians may in part be mediated by Ca2+ balance. We suggest that ion flux may mediate adaptive divergence of natural populations at early life-stages in the face of environmental stress. PMID:26381453

  16. Yeasts Associated with Culex pipiens and Culex theileri Mosquito Larvae and the Effect of Selected Yeast Strains on the Ontogeny of Culex pipiens.

    PubMed

    Steyn, A; Roets, F; Botha, A

    2016-04-01

    The success of mosquitoes in nature has been linked to their microbiota and bacteria in particular. Yet, knowledge on their symbioses with yeasts is lacking. To explore possible associations, culturable yeasts were isolated from wild larvae of Culex pipiens and Culex theileri. These yeasts were classified using restriction fragment length polymorphism (RFLP) analyses and identified by sequencing the D1/D2 region of the 26S rRNA gene. Representative strains of Candida, Cryptococcus, Galactomyces, Hannaella, Meyerozyma, Pichia, Rhodosporidium, Rhodotorula, Trichosporon and Wickerhamomyces were isolated. Our results provide, to our knowledge, the first records of the yeast microbiota from wild mosquito larvae and show that they may harbour potential clinically relevant yeast species, including the well-known opportunistic human pathogen Candida albicans. Also, diminished numbers of yeast isolates originating from adults, compared to larvae, support the hypothesis of microbial reduction/elimination during adult emergence and extend it to include yeasts. In addition, strains of Candida albicans, Candida glabrata, Candida pseudolambica, Cryptococcus gattii, Metschnikowia bicuspidata, Saccharomyces cerevisiae and Wickerhamomyces anomalus were tested as sole feed during a 21-day feeding experiment wherein cumulative larval growth, survival and pupation of Cx. pipiens were recorded. Although most yeasts supported larval growth in a similar manner to the positive control S. cerevisiae strain, the different yeast strains impacted differently on Culex pipiens ontogeny. Notably, survival and pupation of larvae were negatively impacted by a representative strain of the primary pathogen C. gattii - signifying some yeasts to be natural antagonists of mosquitoes. PMID:26573833

  17. Oviposition Activity Patterns of Culex pipiens and Culex restuans in Pennsylvania.

    PubMed

    Stough, Jennifer E; Wallace, John R

    2016-06-01

    Culex pipiens and Cx. restuans are the main vectors of West Nile virus and the primary target species of surveillance and control programs in Pennsylvania. Performing adult control, specifically ultra-low volume (ULV) applications, at night during peak oviposition activity time(s) is necessary to control these species. In July and August of 2009, collections were made at 15-min intervals starting at sunset and continuing until 3 h after sunset to establish a more accurate timeline of Cx. pipiens and Cx. restuans oviposition flight activity. The highest numbers of Cx. pipiens and Cx. restuans were collected during the 15-30, 30-45, and 45-60 min postsunset time intervals (P < 0.05). Oviposition activity began to decrease after 60 min postsunset. These observations have identified a smaller oviposition activity period for Cx. pipiens and Cx. restuans than noted from other studies, thus potentially improving the timing of ULV operations to control these 2 vector species. PMID:27280355

  18. Larvicidal Activity of Nerium oleander against Larvae West Nile Vector Mosquito Culex pipiens (Diptera: Culicidae).

    PubMed

    El-Akhal, Fouad; Guemmouh, Raja; Ez Zoubi, Yassine; El Ouali Lalami, Abdelhakim

    2015-01-01

    Background. Outbreaks of the West Nile virus infection were reported in Morocco in 1996, 2003, and 2010. Culex pipiens was strongly suspected as the vector responsible for transmission. In the North center of Morocco, this species has developed resistance to synthetic insecticides. There is an urgent need to find alternatives to the insecticides as natural biocides. Objective. In this work, the insecticidal activity of the extract of the local plant Nerium oleander, which has never been tested before in the North center of Morocco, was studied on larval stages 3 and 4 of Culex pipiens. Methods. Biological tests were realized according to a methodology inspired from standard World Health Organization protocol. The mortality values were determined after 24 h of exposure and LC50 and LC90 values were calculated. Results. The extract had toxic effects on the larvae of culicid mosquitoes. The ethanolic extract of Nerium oleander applied against the larvae of Culex pipiens has given the lethal concentrations LC50 and LC90 in the order of 57.57 mg/mL and 166.35 mg/mL, respectively. Conclusion. This investigation indicates that N. oleander could serve as a potential larvicidal, effective natural biocide against mosquito larvae, particularly Culex pipiens. PMID:26640701

  19. Mom Matters: Diapause Characteristics of Culex pipiens-Culex quinquefasciatus (Diptera: Culicidae) Hybrid Mosquitoes.

    PubMed

    Meuti, Megan E; Short, Clancy A; Denlinger, David L

    2015-03-01

    Females of the northern house mosquito, Culex pipiens L., are capable of entering an adult overwintering diapause characterized by arrested ovarian development, enhanced stress tolerance, and elevated lipid stores. In contrast, the southern house mosquito, Culex quinquefasciatus Say, lacks this capacity and is therefore unable to survive the harsh winters found in northern regions of North America. These two species are capable of forming fertile hybrids in the United States, yet the diapause characteristics of these hybrids have not been extensively investigated. We crossed Cx. pipiens from Columbus, OH, with Cx. quinquefasciatus from Vero Beach, FL, and reared F1 hybrids from all mothers separately under diapause-inducing, short-day conditions (a photoperiod of 8:16 [L:D] h) at 18°C. Egg follicle length and lipid content were used to assess the diapause status of hybrids. Diapause incidence of hybrids varied widely for progeny from different mothers of the same species, but hybrids with Cx. pipiens mothers were consistently more prone to enter diapause than hybrids that had Cx. quinquefasciatus mothers. Our results suggest a strong maternal influence on the diapause phenotype and that a high percentage (45-75%) of Cx. pipiens-Cx. quinquefasciatus hybrids are capable of entering diapause. This implies that many hybrids can successfully overwinter, leading to a possible widening of the hybrid zone of these two species in North America. PMID:26336296

  20. Contrasting levels of variability between cytoplasmic genomes and incompatibility types in the mosquito Culex pipiens.

    PubMed

    Guillemaud, T; Pasteur, N; Rousset, F

    1997-02-22

    Reproductive incompatibilities called cytoplasmic incompatibilities are known to affect a large number of arthropod species and are mediated by Wolbachia, a maternally transmitted microorganism. The crossing relationships between strains of potential hosts define their incompatibility types and it is generally assumed that differences between strains of Wolbachia induce different crossing types. Among all the described host species, the mosquito, Culex pipiens, displays the greatest variability of cytoplasmic incompatibility crossing types. We analysed mitochondrial and bacterial DNA variability in Culex pipiens in order to investigate some possible causes of incompatibility crossing type variability. We sequenced fragments of the ftsZ gene, and the A + T-rich control region of the mtDNA. We also sequenced the second subunit of the mitochondrial cytochrome oxidase (COII) gene, in Culex pipiens and a closely related species, C. torrentium, in order to verify the usefulness of the A + T-rich region for the present purposes. No variability was found in the Wolbachia ftsZ gene fragment, and very limited variation of the mitochondrial marker whatever the compatibility type or the origin of the host. A low variability was found in the A + T-rich region and comparison of divergence of the A + T-rich region and COII gene between C. pipiens and C. torrentium did not reveal any special constraints affecting this region. In contrast to observations in other host species, variability of incompatibility crossing types is not due to multiple infections by distantly related Wolbachia strains. PMID:9061971

  1. Larvicidal Activity of Nerium oleander against Larvae West Nile Vector Mosquito Culex pipiens (Diptera: Culicidae)

    PubMed Central

    El-Akhal, Fouad; Guemmouh, Raja; Ez Zoubi, Yassine; El Ouali Lalami, Abdelhakim

    2015-01-01

    Background. Outbreaks of the West Nile virus infection were reported in Morocco in 1996, 2003, and 2010. Culex pipiens was strongly suspected as the vector responsible for transmission. In the North center of Morocco, this species has developed resistance to synthetic insecticides. There is an urgent need to find alternatives to the insecticides as natural biocides. Objective. In this work, the insecticidal activity of the extract of the local plant Nerium oleander, which has never been tested before in the North center of Morocco, was studied on larval stages 3 and 4 of Culex pipiens. Methods. Biological tests were realized according to a methodology inspired from standard World Health Organization protocol. The mortality values were determined after 24 h of exposure and LC50 and LC90 values were calculated. Results. The extract had toxic effects on the larvae of culicid mosquitoes. The ethanolic extract of Nerium oleander applied against the larvae of Culex pipiens has given the lethal concentrations LC50 and LC90 in the order of 57.57 mg/mL and 166.35 mg/mL, respectively. Conclusion. This investigation indicates that N. oleander could serve as a potential larvicidal, effective natural biocide against mosquito larvae, particularly Culex pipiens. PMID:26640701

  2. Blood-feeding patterns of the Culex pipiens complex in Sacramento and Yolo Counties, California.

    PubMed

    Montgomery, Matthew J; Thiemann, Tara; Macedo, Paula; Brown, David A; Scott, Thomas W

    2011-03-01

    Mosquitoes in the Culex pipiens complex are competent vectors of West Nile virus (WNV; family Flaviviridae, genus Flavivirus) in the laboratory, and field-collected mosquitoes have tested positive for the virus in California and elsewhere. A better understanding of Cx. pipiens complex blood-feeding patterns will help define the threat that these mosquitoes pose to human health and their role in WNV amplification in northern California. We collected blood-engorged Cx. pipiens complex mosquitoes from resting sites near and away from human habitation in Sacramento and Yolo Counties. Cytochrome c oxidase 1 gene sequences were used to identify the vertebrate species from which blood meals were taken. Of 330 engorged mosquitoes collected at 28 sites from June through August 2007 and May through August 2008, >99% fed on an avian host. Three mosquitoes contained bovine blood and none had fed on a human. American Robins (Turdus migratorius) were bitten most often, and the proportion of American Robin blood meals increased significantly over the summer. Other important avian hosts included House Finches (Carpodacus mexicanus), Barn Swallows (Hirundo rustica), Western Meadowlarks (Sturnella neglecta), and Mourning Doves (Zenaida macroura). In rural areas, Barn Swallows, Brewer's Blackbirds (Euphagus cyanocephalus), and House Sparrows (Passer domesticus) were frequent hosts. In settings near human habitation, Mourning Doves and Western Meadowlarks were common hosts. Our data indicate that in north central California mosquitoes in the Cx. pipiens complex may be more important as epiornitic than epidemic vectors of WNV. PMID:21485380

  3. [The spatial distribution and infection of Culex pipiens L. mosquitoes with Western Nile virus in the Volgograd region].

    PubMed

    Fedorova, M V; Borodaĭ, N V; Shaĭkevich, E V

    2015-01-01

    The infection of Culex pipiens f. pipiens and Cx. pipiens f. molestus mosquitoes with Western Nile virus was studied. Their taxonomic status was determined from the type of mitochondrial DNA. Among 424 females individually examined, there were 4 infected specimens: 3 pipens females gathered in the urban and suburban open biotopes and one molestus female caught in the open urban biotope. Molestus mitotype females were first detected in the suburban open biotopes. The possible reasons for the wide colonization of molestus mosquitoes and its epidemic consequences are discussed. PMID:25850309

  4. Leopard frog and wood frog reproduction in Colorado and Wyoming

    USGS Publications Warehouse

    Corn, Paul Stephen; Livo, Lauren J.

    1989-01-01

    Between 1978 and 1988, we recorded reproductive information from populations of ranid frogs in Colorado and Wyoming. Egg masses from five plains and montane populations of northern leopard frogs (Rana pipiens) contained 645-6272 eggs (x̄ = 3045, N = 68 egg masses). In two montane populations of wood frogs (Rana sylvatica) numbers of eggs per egg mass varied from 711-1248 (x̄ = 876, N = 15) and probably were equal to total clutch size. Mean hatching success was 90% in egg masses from one R. sylvatica population and ranged from 70% to 99% in R. pipiens egg masses. Rana pipiens egg masses from one location were assigned to three overlapping size distributions, which we believe reflects the underlying age structure of female frogs.

  5. Widespread occurrence of the chytrid fungus batrachochytrium dendrobatidis on oregon spotted frogs (rana pretiosa)

    USGS Publications Warehouse

    Pearl, C.A.; Bowerman, J.; Adams, M.J.; Chelgren, N.D.

    2009-01-01

    The pathogen Batrachochytrium dendrobatidis (Bd) has been associated with amphibian declines in multiple continents, including western North America. We investigated Bd prevalence in Oregon spotted frog (Rana pretiosa), a species that has declined across its range in the Pacific Northwest. Polymerase chain reaction analysis of skin swabs indicated that Bd was prevalent within populations (420 of 617 juvenile and adults) and widespread among populations (36 of 36 sites) where we sampled R. pretiosa in Oregon and Washington. We rarely detected Bd in R. pretiosa larvae (2 of 72). Prevalence of Bd in postmetamorphic R. pretiosa was inversely related to frog size. We found support for an interactive effect of elevation and sampling date on Bd: prevalence of Bd generally increased with date, but this effect was more pronounced at lower elevations. We also found evidence that the body condition of juvenile R. pretiosa with Bd decreased after their first winter. Our data indicate that some Oregon spotted frog populations are currently persisting with relatively high Bd prevalence, but the risk posed by Bd is unknown. ?? 2010 International Association for Ecology and Health.

  6. "Bird biting" mosquitoes and human disease: a review of the role of Culex pipiens complex mosquitoes in epidemiology.

    PubMed

    Farajollahi, Ary; Fonseca, Dina M; Kramer, Laura D; Marm Kilpatrick, A

    2011-10-01

    The transmission of vector-borne pathogens is greatly influenced by the ecology of their vector, which is in turn shaped by genetic ancestry, the environment, and the hosts that are fed on. One group of vectors, the mosquitoes in the Culex pipiens complex, play key roles in the transmission of a range of pathogens including several viruses such as West Nile and St. Louis encephalitis viruses, avian malaria (Plasmodium spp.), and filarial worms. The Cx. pipiens complex includes Culex pipiens pipiens with two forms, pipiens and molestus, Culex pipiens pallens, Culex quinquefasciatus, Culex australicus, and Culex globocoxitus. While several members of the complex have limited geographic distributions, Cx. pipienspipiens and Cx. quinquefasciatus are found in all known urban and sub-urban temperate and tropical regions, respectively, across the world, where they are often principal disease vectors. In addition, hybrids are common in areas of overlap. Although gaps in our knowledge still remain, the advent of genetic tools has greatly enhanced our understanding of the history of speciation, domestication, dispersal, and hybridization. We review the taxonomy, genetics, evolution, behavior, and ecology of members of the Cx. pipiens complex and their role in the transmission of medically important pathogens. The adaptation of Cx. pipiens complex mosquitoes to human-altered environments led to their global distribution through dispersal via humans and, combined with their mixed feeding patterns on birds and mammals (including humans), increased the transmission of several avian pathogens to humans. We highlight several unanswered questions that will increase our ability to control diseases transmitted by these mosquitoes. PMID:21875691

  7. Chemical composition and larvicidal evaluation of Mentha, Salvia, and Melissa essential oils against the West Nile virus mosquito Culex pipiens.

    PubMed

    Koliopoulos, George; Pitarokili, Danae; Kioulos, Elias; Michaelakis, Antonios; Tzakou, Olga

    2010-07-01

    The volatile metabolites of wild-growing Mentha spicata, M. longifolia, M. suaveolens, Melissa officinalis, Salvia fruticosa, S. pomifera subsp. calycina, and S. pomifera subsp. pomifera from Greece were determined by gas chromatography and gas chromatography-mass spectrometry. The insecticidal properties of the analyzed essential oils were screened on Culex pipiens larvae. Additionally two of the main components of the essential oils, piperitenone oxide and 1,8-cineole were assayed against C. pipiens in order to define the affiliation between them and the larvicidal properties of the oils. The most effective oils were M. suaveolens (major constituent piperitenone oxide, 62.4%), M. spicata (piperitenone oxide, 35.7% and 1,8-cineole, 14.5%) and M. longifolia--Central Greece (piperitenone oxide, 33.4%; 1,8-cineole, 24.5% and trans-piperitone epoxide, 17.4%), which exhibited LC(50) values ranging from 47.88 to 59.33 mg l(-1). Medium activity revealed the oils of M. officinalis (terpin-4-ol, 15.8%; caryophyllene oxide, 13.2%; sabinene, 12.9%; beta-pinene, 12.1%; and trans-caryophyllene, 10.2%), M. longifolia--Southern Greece (carvone, 54.7% and limonene 20.0%), S. pomifera subsp. pomifera (trans-caryophyllene, 22.5% and trans-thujone, 21.0%), S. pomifera subsp. calycina--West Southern Greece (trans-thujone, 56.1% and 1,8-cineole, 10.4%), and S. fruticosa--population 2 (camphor, 23.1%; alpha-pinene, 12.7%; and borneol, 12.6%), with LC(50) values ranging from 78.28 to 91.45 mg l(-1). S. pomifera subsp. calycina (Central Greece) essential oil (trans-thujone, 26.5% and cis-thujone, 12.0%) presented rather low activity (LC(50) values 140.42 mg l(-1)), while S. fruticosa--population 1 (1,8-cineole, 31.4% and camphor, 22.6%) was the only inactive oil. Additionally, the constituent piperitenone oxide was found to be highly active (LC(50) values 9.95 mg l(-1)), whereas 1,8-cineole revealed no toxicity. PMID:20405142

  8. Range-wide phylogeographic analysis of the spotted frog complex (Rana luteiventris and Rana pretiosa) in northwestern North America

    USGS Publications Warehouse

    Funk, W.C.; Pearl, C.A.; Draheim, H.M.; Adams, M.J.; Mullins, T.D.; Haig, S.M.

    2008-01-01

    The dynamic geological and climatic history of northwestern North America has made it a focal region for phylogeography. We conducted a range-wide phylogeographic analysis of the spotted frog complex (Rana luteiventris and Rana pretiosa) across its range in northwestern North America to understand its evolutionary history and the distribution of clades to inform conservation of R. pretiosa and Great Basin R. luteiventris, candidates for listing under the US Endangered Species Act. Mitochondrial DNA sequence data from a segment of the cytochrome b gene were obtained from 308 R. luteiventris and R. pretiosa from 96 sites. Phylogenetic analysis revealed one main R. pretiosa clade and three main R. luteiventris clades, two of which overlapped in southeastern Oregon. The three R. luteiventris clades were separated from each other by high levels of sequence divergence (average of 4.75-4.97%). Two divergent clades were also uncovered within the Great Basin. Low genetic variation in R. pretiosa and the southeastern Oregon clade of R. luteiventris suggests concern about their vulnerability to extinction. ?? 2008 Elsevier Inc.

  9. Culex pipiens as a potential vector for transmission of Dirofilaria immitis and other unclassified Filarioidea in Southwest Spain.

    PubMed

    Bravo-Barriga, Daniel; Parreira, Ricardo; Almeida, António P G; Calado, Manuela; Blanco-Ciudad, Juan; Serrano-Aguilera, Francisco Javier; Pérez-Martín, Juan Enrique; Sánchez-Peinado, Joaquín; Pinto, João; Reina, David; Frontera, Eva

    2016-06-15

    Dirofilaria immitis is one of the most frequently detected mosquito-transmitted zoonotic filarioid nematode in mammals in Europe, being canine dirofilariosis a major animal health problem, endemic in the Mediterranean area. This study, focused on Southwest Spain, in order to bring new insights into (i) the epidemiology of Dirofilaria spp., (ii) the species of Culicid vectors possibly involved in their transmission and (iii) the genetic variability of those potential vectors. A total of 881 adult female mosquitoes from 11 different species, were captured during 2012-2013, and detection of filarioid DNA was attempted by PCR using specific primers (ITS-2 and COI), followed by DNA sequencing. In a single Culex pipiens specimen D. immitis DNA was detected both in the head-thorax and abdomen sections. Filarioid nematode DNA was also detected in eight additional Cx. pipiens specimens also in both the thorax and the abdomen, but analysis of sequence data did not allow unambiguous assignment of any of the obtained sequences to a previously defined species. All Cx. pipiens with filarioid DNA were individually analysed by CQ11 to discriminate between pipiens, molestus, and hybrid forms. Besides, rDNA ITS-2 sequence analysis revealed the presence of haplotype H1 and H2 of Cx. pipiens. To our knowledge this study revealed, for the first time in Spain, the occurrence of likely mature infection of D. immitis in Cx. pipiens, as well as with other yet uncharacterized nematodes, supporting its role as a potential vector of these filarids. PMID:27198797

  10. The precarious persistence of the endangered Sierra Madre yellow-legged frog Rana muscosa in southern California, USA

    USGS Publications Warehouse

    Backlin, Adam R.; Hitchcock, Cynthia J.; Gallegos, Elizabeth A.; Yee, Julie L.; Fisher, Robert N.

    2015-01-01

    We conducted surveys for the Endangered Sierra Madre yellow-legged frog Rana muscosa throughout southern California to evaluate the current distribution and status of the species. Surveys were conducted during 2000–2009 at 150 unique streams and lakes within the San Gabriel, San Bernardino, San Jacinto, and Palomar mountains of southern California. Only nine small, geographically isolated populations were detected across the four mountain ranges, and all tested positive for the amphibian chytrid fungus Batrachochytrium dendrobatidis. Our data show that when R. muscosa is known to be present it is easily detectable (89%) in a single visit during the frog's active season. We estimate that only 166 adult frogs remained in the wild in 2009. Our research indicates that R. muscosa populations in southern California are threatened by natural and stochastic events and may become extirpated in the near future unless there is some intervention to save them.

  11. Early warning of West Nile virus mosquito vector: climate and land use models successfully explain phenology and abundance of Culex pipiens mosquitoes in north-western Italy

    PubMed Central

    2014-01-01

    Background West Nile Virus (WNV) is an emerging global health threat. Transmission risk is strongly related to the abundance of mosquito vectors, typically Culex pipiens in Europe. Early-warning predictors of mosquito population dynamics would therefore help guide entomological surveillance and thereby facilitate early warnings of transmission risk. Methods We analysed an 11-year time series (2001 to 2011) of Cx. pipiens mosquito captures from the Piedmont region of north-western Italy to determine the principal drivers of mosquito population dynamics. Linear mixed models were implemented to examine the relationship between Cx. pipiens population dynamics and environmental predictors including temperature, precipitation, Normalized Difference Water Index (NDWI) and the proximity of mosquito traps to urban areas and rice fields. Results Warm temperatures early in the year were associated with an earlier start to the mosquito season and increased season length, and later in the year, with decreased abundance. Early precipitation delayed the start and shortened the length of the mosquito season, but increased total abundance. Conversely, precipitation later in the year was associated with a longer season. Finally, higher NDWI early in the year was associated with an earlier start to the season and increased season length, but was not associated with abundance. Proximity to rice fields predicted higher total abundance when included in some models, but was not a significant predictor of phenology. Proximity to urban areas was not a significant predictor in any of our models. Predicted variations in start of the season and season length ranged from one to three weeks, across the measured range of variables. Predicted mosquito abundance was highly variable, with numbers in excess of 1000 per trap per year when late season temperatures were low (average 21°C) to only 150 when late season temperatures were high (average 30°C). Conclusions Climate data collected early in

  12. Genetic divergence between populations of feral and domestic forms of a mosquito disease vector assessed by transcriptomics

    PubMed Central

    2015-01-01

    Culex pipiens, an invasive mosquito and vector of West Nile virus in the US, has two morphologically indistinguishable forms that differ dramatically in behavior and physiology. Cx. pipiens form pipiens is primarily a bird-feeding temperate mosquito, while the sub-tropical Cx. pipiens form molestus thrives in sewers and feeds on mammals. Because the feral form can diapause during the cold winters but the domestic form cannot, the two Cx. pipiens forms are allopatric in northern Europe and, although viable, hybrids are rare. Cx. pipiens form molestus has spread across all inhabited continents and hybrids of the two forms are common in the US. Here we elucidate the genes and gene families with the greatest divergence rates between these phenotypically diverged mosquito populations, and discuss them in light of their potential biological and ecological effects. After generating and assembling novel transcriptome data for each population, we performed pairwise tests for nonsynonymous divergence (Ka) of homologous coding sequences and examined gene ontology terms that were statistically over-represented in those sequences with the greatest divergence rates. We identified genes involved in digestion (serine endopeptidases), innate immunity (fibrinogens and α-macroglobulins), hemostasis (D7 salivary proteins), olfaction (odorant binding proteins) and chitin binding (peritrophic matrix proteins). By examining molecular divergence between closely related yet phenotypically divergent forms of the same species, our results provide insights into the identity of rapidly-evolving genes between incipient species. Additionally, we found that families of signal transducers, ATP synthases and transcription regulators remained identical at the amino acid level, thus constituting conserved components of the Cx. pipiens proteome. We provide a reference with which to gauge the divergence reported in this analysis by performing a comparison of transcriptome sequences from conspecific

  13. Genetic divergence between populations of feral and domestic forms of a mosquito disease vector assessed by transcriptomics.

    PubMed

    Price, Dana C; Fonseca, Dina M

    2015-01-01

    Culex pipiens, an invasive mosquito and vector of West Nile virus in the US, has two morphologically indistinguishable forms that differ dramatically in behavior and physiology. Cx. pipiens form pipiens is primarily a bird-feeding temperate mosquito, while the sub-tropical Cx. pipiens form molestus thrives in sewers and feeds on mammals. Because the feral form can diapause during the cold winters but the domestic form cannot, the two Cx. pipiens forms are allopatric in northern Europe and, although viable, hybrids are rare. Cx. pipiens form molestus has spread across all inhabited continents and hybrids of the two forms are common in the US. Here we elucidate the genes and gene families with the greatest divergence rates between these phenotypically diverged mosquito populations, and discuss them in light of their potential biological and ecological effects. After generating and assembling novel transcriptome data for each population, we performed pairwise tests for nonsynonymous divergence (Ka) of homologous coding sequences and examined gene ontology terms that were statistically over-represented in those sequences with the greatest divergence rates. We identified genes involved in digestion (serine endopeptidases), innate immunity (fibrinogens and α-macroglobulins), hemostasis (D7 salivary proteins), olfaction (odorant binding proteins) and chitin binding (peritrophic matrix proteins). By examining molecular divergence between closely related yet phenotypically divergent forms of the same species, our results provide insights into the identity of rapidly-evolving genes between incipient species. Additionally, we found that families of signal transducers, ATP synthases and transcription regulators remained identical at the amino acid level, thus constituting conserved components of the Cx. pipiens proteome. We provide a reference with which to gauge the divergence reported in this analysis by performing a comparison of transcriptome sequences from conspecific

  14. Does testis weight decline towards the Subarctic? A case study on the common frog, Rana temporaria

    NASA Astrophysics Data System (ADS)

    Hettyey, Attila; Laurila, Anssi; Herczeg, Gábor; Jönsson, K. Ingemar; Kovács, Tibor; Merilä, Juha

    2005-04-01

    Interpopulation comparisons of variation in resource availability and in allocation patterns along altitudinal and latitudinal gradients allow insights into the mechanisms shaping the life history of animals. Patterns of between-population differences in female life history traits have been studied intensively across a wide range of taxa, but similar investigations in males have remained scarce. To study if testis weight—a measure of reproductive investment—varies on a geographical scale in anurans, we focussed on the variation in relative testis weight (RelTW) and asymmetry in 22 populations of the common frog Rana temporaria along a 1,600-km latitudinal transect across the Scandinavian peninsula. We found that RelTW decreased towards the north. Body mass and body length both had independent positive effects on testes mass. We found evidence for directional asymmetry (DA) in testis weight with the right testis being larger than the left. The level of DA in testis weight was not related to latitude, but both body mass and testes mass had independent positive effects on asymmetry. We discuss the northwards decrease in RelTW in terms of a decreased reproductive investment as a possible consequence of harsher environmental conditions, and perhaps also, weaker sexual selection in the north than in the south.

  15. Spontaneous otoacoustic emissions in the European edible frog (Rana esculenta): spectral details and temperature dependence.

    PubMed

    van Dijk, P; Wit, H P; Segenhout, J M

    1989-11-01

    Spontaneous otoacoustic emissions were recorded in 41 ears of 29 European edible frogs (Rana esculenta). Emission frequencies ranged from 450 to 1350 Hz. The distribution of frequencies shows two distinct populations: one above and one below 1 kHz. With one exception, a maximum number of two emissions were recorded per ear, each in a different population. An amplitude distribution of a frog emission was sampled, from which it was concluded that the emission is generated by an active oscillator. The spectral width of an emission ranged from 1 to 200 Hz (average 38 Hz). There was negative correlation between sound pressure level of an emission and spectral width. In 4 frogs the dependence of emission power and frequency on temperature was investigated. An emission could be 'switched on and off' within a few degrees centigrade. At temperatures below the switching interval no emission was recorded; for higher temperatures emission power showed no dependence on temperature. Frequency increased with temperature (Q10 = 1.1 to 1.3). This yields a mismatch with temperature dependence of best frequencies of auditory fibers. The consequences of this mismatch are discussed. PMID:2691473

  16. Larvicidal activity of two Algerian Verbenaceae essential oils against Culex pipiens.

    PubMed

    Zoubiri, Safia; Baaliouamer, Aoumeur

    2011-09-27

    The purpose of this study was to evaluate the bioactivity of essential oils extracted from the leaves of Verbena officinalis and Lantana camara L. for the control of Culex pipiens. Triplicate bioassays were performed with fourth larval instars of C. pipiens (n=25 per replicate) with solutions at 1, 5, 10, 50, 100 and 500 μL/L of V. officinalis and L. camara L. extracts. Cumulative mortalities were determined 3, 6, 12 and 24h after treatment. Results showed cumulative mortalities, at three (3) hours to be 30.0 ± 2.9% and 14.8 ± 1.5% and achieve 43.3 ± 1.9% and 44.4 ± 3.1% after 24h exposure time, at 100mg/L of essential oil from L. camara and V. officinalis, respectively. PMID:21592668

  17. Repellency and toxicity of aromatic plant extracts against the mosquito Culex pipiens molestus (Diptera: Culicidae).

    PubMed

    Traboulsi, Abdallah F; El-Haj, Samih; Tueni, Marie; Taoubi, Khalil; Nader, Natalie Abi; Mrad, Abir

    2005-06-01

    The insecticidal activities of essential oil extracts from leaves, flowers and roots of aromatic plants against fourth-instar larvae of the mosquito Culex pipiens molestus Forskal were determined. Extracts of Foeniculum vulgare Mill were the most toxic, followed by those of Ferula hermonis Boiss, Citrus sinensis Osbeck, Pinus pinea L, Laurus nobilis L and Eucalyptus spp with LC50 values of 24.5, 44.0, 60.0, 75.0, 117.0 and 120.0 mg litre(-1), respectively. Combination tests between the LC50 and the maximum sub-lethal concentration (MSLC) were determined. Over 20 major components were identified in extracts from each plant species tested. Five essential oils and nine pure components were studied for their repellency against mosquito bites. Terpineol and 1,8-cineole were the most effective against Culex pipiens molestus bites offering complete protection for 1.6 and 2 h, respectively. PMID:15662650

  18. Extraction and antioxidant activity of polysaccharides from Rana chensinensis skin.

    PubMed

    Wang, Zhanyong; Zhao, Yuanyuan; Su, Tingting

    2015-01-22

    The extraction process of polysaccharides from Rana chensinensis skin was optimized by using a Box-Behnken design. The optimum extraction conditions were as follows: extraction time, 4.96h; extraction temperature, 100°C; ratio of water to raw material, 60; and extraction frequency, 1. Under these conditions, the experimental polysaccharide yield was 2.03±0.14%, which agreed with the predicted yield. The purified polysaccharide RCSP II was successfully obtained by diethylaminoethanol-Sepharose and Sepharose CL-6B column chromatography. In vitro experiments showed that RCSP II exhibited a strong scavenging activity against superoxide anion and 1,1-diphenyl-2-picrylhydrazyl radicals but a weak scavenging activity against hydroxyl radicals. RCSP II also showed a strong reducing capacity. Thus, this polysaccharide can be used as a natural antioxidant in functional foods or medicines. PMID:25439863

  19. [Origin of the differences of superficial potentials in Rana esculenta].

    PubMed

    Fontas, B; Mambrini, J

    1977-07-18

    The spatial distributions of superficial D.C. potentials on the skin of Rana esculenta have been compared to those of the intensity of short-circuit current (S.C.C.) expressing the transcutaneous active transport of sodium ions. It has been observed that the sites of maximum D.C. potentials coincide with the localisations of maximum S.C.C. values. Moreover, superficial D.C. potentials and S.C.C. are similarly modified by the depression of metabolic activity due to lowered temperature or poisoning by dinitrophenol (DNP). It is thus proposed that the spatial distribution of the transcutaneous active transport system for sodium ions is the origin of the electric generator of superficial D.C. potentials. PMID:409557

  20. Adaptive divergence of the moor frog (Rana arvalis) along an acidification gradient

    PubMed Central

    2011-01-01

    Background Environmental stress can result in strong ecological and evolutionary effects on natural populations, but to what extent it drives adaptive divergence of natural populations is little explored. We used common garden experiments to study adaptive divergence in embryonic and larval fitness traits (embryonic survival, larval growth, and age and size at metamorphosis) in eight moor frog, Rana arvalis, populations inhabiting an acidification gradient (breeding pond pH 4.0 to 7.5) in southwestern Sweden. Embryos were raised until hatching at three (pH 4.0, 4.3 and 7.5) and larvae until metamorphosis at two (pH 4.3 and 7.5) pH treatments. To get insight into the putative selective agents along this environmental gradient, we measured relevant abiotic and biotic environmental variables from each breeding pond, and used linear models to test for phenotype-environment correlations. Results We found that acid origin populations had higher embryonic and larval acid tolerance (survival and larval period were less negatively affected by low pH), higher larval growth but slower larval development rates, and metamorphosed at a larger size. The phenotype-environment correlations revealed that divergence in embryonic acid tolerance and metamorphic size correlated most strongly with breeding pond pH, whereas divergence in larval period and larval growth correlated most strongly with latitude and predator density, respectively. Conclusion Our results suggest that R. arvalis has diverged in response to pH mediated selection along this acidification gradient. However, as latitude and pH were closely spatially correlated in this study, further studies are needed to disentangle the specific agents of natural selection along acidification gradients. Our study highlights the need to consider the multiple interacting selective forces that drive adaptive divergence of natural populations along environmental stress gradients. PMID:22182445

  1. Genome Evolution of Wolbachia Strain wPip from the Culex pipiens Group

    PubMed Central

    Klasson, Lisa; Walker, Thomas; Sebaihia, Mohammed; Sanders, Mandy J.; Quail, Michael A.; Lord, Angela; Sanders, Susanne; Earl, Julie; O'Neill, Scott L.; Thomson, Nicholas; Sinkins, Steven P.; Parkhill, Julian

    2008-01-01

    The obligate intracellular bacterium Wolbachia pipientis strain wPip induces cytoplasmic incompatibility (CI), patterns of crossing sterility, in the Culex pipiens group of mosquitoes. The complete sequence is presented of the 1.48-Mbp genome of wPip which encodes 1386 coding sequences (CDSs), representing the first genome sequence of a B-supergroup Wolbachia. Comparisons were made with the smaller genomes of Wolbachia strains wMel of Drosophila melanogaster, an A-supergroup Wolbachia that is also a CI inducer, and wBm, a mutualist of Brugia malayi nematodes that belongs to the D-supergroup of Wolbachia. Despite extensive gene order rearrangement, a core set of Wolbachia genes shared between the 3 genomes can be identified and contrasts with a flexible gene pool where rapid evolution has taken place. There are much more extensive prophage and ankyrin repeat encoding (ANK) gene components of the wPip genome compared with wMel and wBm, and both are likely to be of considerable importance in wPip biology. Five WO-B–like prophage regions are present and contain some genes that are identical or highly similar in multiple prophage copies, whereas other genes are unique, and it is likely that extensive recombination, duplication, and insertion have occurred between copies. A much larger number of genes encode ankyrin repeat (ANK) proteins in wPip, with 60 present compared with 23 in wMel, many of which are within or close to the prophage regions. It is likely that this pattern is partly a result of expansions in the wPip lineage, due for example to gene duplication, but their presence is in some cases more ancient. The wPip genome underlines the considerable evolutionary flexibility of Wolbachia, providing clear evidence for the rapid evolution of ANK-encoding genes and of prophage regions. This host–Wolbachia system, with its complex patterns of sterility induced between populations, now provides an excellent model for unraveling the molecular systems underlying host

  2. Evaluation of Isotope 32P Method to Mark Culex pipiens (Diptera: Culicidae) in a Laboratory

    PubMed Central

    Zhang, Chongxing; Shi, Guihong; Zhao, Yuqiang; Yan, Dongmei; Li, Huaiju; Liu, Hongmei; Wiwatanaratanabutr, Itsanun; Gong, Maoqing

    2016-01-01

    Background: The aim of the current study was to develop a marking technique as an internal marker to mark post blood meal mosquitoes by using stable phosphate isotope 32P and determine the optimal concentration of it. Methods: An isotonic physiological saline solution, containing different concentration of radioactive isotope 32P-labeled disodium phosphate (Na2H32PO4) was injected into rabbits via the jugular vein in the laboratory. Emerged Cx. pipiens were marked after feeding on rabbit. At the same time, the labeled conditions of emerged Cx. pipiens were also measured by placing feces of No. 6 rabbit into containers with mosquito larvae and pupae inside. Results: According to the label condition of Cx. pipiens after taking blood and the effect of different dosage Na2H32PO4 on rabbit health, the optimal concentration of radioactive isotope was determined, that is, 0.1211 mCi/kg. By placing feces of No. 6 rabbit into containers with mosquito larvae and pupae inside, the emerged mosquitoes were also labeled. Therefore, feeding mosquitoes on the animal injected with radioactive Na2H32PO4 was more practical for detecting and tracing mosquitoes. Conclusion: The method was less time-consuming, more sensitive and safer. This marking method will facilitate post-bloodmeal studies of mosquitoes and other blood-sucking insects. PMID:27308279

  3. GENE-dosage effects on fitness in recent adaptive duplications: ace-1 in the mosquito Culex pipiens.

    PubMed

    Labbé, Pierrick; Milesi, Pascal; Yébakima, André; Pasteur, Nicole; Weill, Mylène; Lenormand, Thomas

    2014-07-01

    Gene duplications have long been advocated to contribute to the evolution of new functions. The role of selection in their early spread is more controversial. Unless duplications are favored for a direct benefit of increased expression, they are likely detrimental. In this article, we investigated the case of duplications favored because they combine already functionally divergent alleles. Their gene-dosage/fitness relations are poorly known because selection may operate on both overall expression and duplicates relative dosage. Using the well-documented case of Culex pipiens resistance to insecticides, we compared strains with various ace-1 allele combinations, including two duplicated alleles carrying both susceptible and resistant copies. The overall protein activity was nearly additive, but, surprisingly, fitness correlated better with the relative proportion of susceptible and resistant copies rather than any absolute measure of activity. Gene dosage is thus crucial, duplications stabilizing a "heterozygote" phenotype. It corroborates the view that these were favored because they fix a permanent heterosis, thereby solving the irreducible trade-off between resistance and synaptic transmission. Moreover, we showed that the contrasted successes of the two duplicated alleles in natural populations depend on genetic changes unrelated to ace-1, confirming the probable implication of recessive sublethal mutations linked to structural rearrangements in some duplications. PMID:24494966

  4. California red-legged frog (Rana draytonii) movement and habitat use: Implications for conservation

    USGS Publications Warehouse

    Fellers, G.M.; Kleeman, P.M.

    2007-01-01

    Nonbreeding habitats are critically important for Rana draytonii, especially for individuals that breed in temporary bodies of water. We radiotracked 123 frogs to evaluate seasonal habitat use. Individual frogs were continuously tracked for up to 16 months. Some individuals remained at breeding ponds all year, but 66% of female and 25% of male frogs moved to nonbreeding areas, even when the breeding site retained water. Frogs at our main study site moved 150 m (median), roughly the distance to the nearest suitable nonbreeding area. The greatest straight-line distance traveled was 1.4 km, although the presumed distance traveled was 2.8 km. Females were more likely than males to move from permanent ponds (38% of females, 16% of males), but among dispersing frogs, males and females did not differ in distance moved. Some frogs left breeding sites shortly after oviposition (median = 12 days for females, 42.5 days for males), but many individuals remained until the site was nearly dry. Fog provided moisture for dispersal or migration throughout the summer. Our data demonstrate that maintaining populations of pond-breeding amphibians requires that all essential habitat components be protected; these include (1) breeding habitat, (2) nonbreeding habitat, and (3) migration corridors. In addition, a buffer is needed around all three areas to ensure that outside activities do not degrade any of the three habitat components. Copyright 2007 Society for the Study of Amphibians and Reptiles.

  5. Cadmium-induced oxidative stress and apoptosis in the testes of frog Rana limnocharis.

    PubMed

    Zhang, Hangjun; Cai, Chenchen; Shi, Cailei; Cao, Hui; Han, Ziliu; Jia, Xiuying

    2012-10-15

    This study explored the genetic damage induced by cadmium exposure in the testes of Rana limnocharis. Healthy adult frogs were exposed to 2.5, 5, 7.5, or 10 mg/L of cadmium solution for 14 days. The results showed that exposure to these concentrations increased the levels of reactive oxygen species and malondialdehyde content in the testes, clearly indicating a dose-effect relationship. Moreover, the same dosages of Cd(2+) solution increased glutathione (reduced) content, with the values being significantly different from those observed in the control group (P<0.01). The comet assay results demonstrated that the DNA damage rate, tail length, and tail moment of samples obtained from frogs exposed to 2.5-7.5 mg/L of cadmium solution significantly increased compared with those of samples obtained from the control group (P<0.01). These findings suggest that cadmium can induce free radical generation, followed by lipid peroxidation and DNA damage. Ultrastructural observation revealed vacuoles in the spermatogenic cells, cell dispersion, incomplete cell structures, and deformed nucleoli. Moreover, cadmium exposure induced significant down-regulation of Bcl-2 expression and up-regulation of Bax and caspase-3 expressions. Taken together, these data indicate that cadmium can induce testicular cell apoptosis in R. limnocharis. Exploring the effects of cadmium on the mechanism of reproductive toxicity in amphibians will help provide a scientific basis accounting for the global population decline in amphibian species. PMID:22728207

  6. Is body size of the water frog Rana esculenta complex responding to climate change?

    NASA Astrophysics Data System (ADS)

    Tryjanowski, Piotr; Sparks, Tim; Rybacki, Mariusz; Berger, Leszek

    2006-03-01

    Recent studies on climate responses in ectothermic (cold-blooded) vertebrates have been few in number and focussed on phenology rather than morphology. According to Bergmann’s rule, endothermic (warm-blooded) vertebrates from cooler climates tend to be larger than congeners from warmer regions. Although amphibians are ectothermic vertebrates, weather and climatic conditions may also impact on their morphology, and thereby affect their survival rates and population dynamics. In this paper, we show, in a unique long-term study during the period 1963-2003 in an agricultural landscape in western Poland, that the body length of two water frog parental species (males of both Rana ridibunda and R. lessonae) increased significantly. However, their hybridogenetic hybrid R. esculenta did not show similar changes. A significant relationship with a large-scale climatic factor, the winter North Atlantic Oscillation index, was found positive for R. ridibunda males and R. lessonae females, and negative for R. esculenta females. Our findings, the first for amphibians, are consistent with other studies reporting that recent climate change has affected the morphology of animals. However, we also show that changes in amphibian phenotype linked to climate may vary independently between (even very similar) species.

  7. Multiple sublethal chemicals negatively affect tadpoles of the green frog, Rana clamitans

    USGS Publications Warehouse

    Boone, Michelle D.; Bridges, Christine M.; Fairchild, James F.; Little, Edward E.

    2005-01-01

    Many habitats may be exposed to multiple chemical contaminants, particularly in agricultural areas where fertilizer and pesticide use are common; however, the singular and interactive effects of contaminants are not well understood. The objective of our study was to examine how realistic, sublethal environmental levels of ammonium nitrate fertilizer (0, 10, 20 mg/L and ammonium chloride control) and the common insecticide carbaryl (0 or 2.5 mg/L) individually and interactively affect the development, size, and survival of green frog (Rana clamitans) tadpoles. We reared tadpoles for 95 d in outdoor 1,000-L polyethylene ponds. We found that the combination of carbaryl and nitrate had a negative effect on development and mass of tadpoles compared to the positive effect that either contaminant had alone. Presence of carbaryl was generally associated with short-term increases in algal resources, including ponds exposed to both carbaryl and nitrate. However, with exposure to nitrate and carbaryl, tadpole mass and development were not positively affected as with one chemical stressor alone. The combination of these sublethal contaminants may reduce the ability of amphibians to benefit from food-rich environments or have metabolic costs. Our study demonstrates the importance of considering multiple stressors when evaluating population-level responses.

  8. Effects of carbaryl on green frog (Rana clamitans) tadpoles: Timing of exposure versus multiple exposures

    USGS Publications Warehouse

    Boone, M.D.; Bridges, C.M.

    2003-01-01

    The majority of studies on pesticide impacts have evaluated the effects of single exposures. However, multiple exposures to a pesticide may be more prevalent. The objective of our study was to determine how multiple exposures versus single exposure at different times during development affected survival to metamorphosis, tadpole survival, tadpole mass, and tadpole developmental stage of green frog (Rana clamitans) tadpoles reared at low and high density in outdoor cattle tank ponds. Tadpoles were exposed to carbaryl zero, one, two, or three times at 14-d intervals. We applied single doses of carbaryl at one of three times, specifically during early, mid, or late development. Overall, we found that multiple exposures had a greater impact than single exposures during development. More individuals reached metamorphosis in ponds exposed to multiple doses of carbaryl compared with controls, indicating that the presence of carbaryl stimulated metamorphosis. The presence of carbaryl in the aquatic environment also resulted in more developed tadpoles compared with controls. Tadpoles in control ponds did not reach metamorphosis and were less developed than individuals exposed to carbaryl; this effect indicates that, under ideal conditions, green frogs could overwinter in ponds so that greater size could be attained before metamorphosis in the following spring or summer. Our study demonstrated the importance of including realistic application procedures when evaluating the effects of a pesticide and that multiple exposures to a short-lived pesticide are more likely to affect an amphibian population.

  9. Cytoplasmic incompatibility as a means of controlling Culex pipiens quinquefasciatus mosquito in the islands of the south-western Indian Ocean.

    PubMed

    Atyame, Célestine M; Pasteur, Nicole; Dumas, Emilie; Tortosa, Pablo; Tantely, Michaël Luciano; Pocquet, Nicolas; Licciardi, Séverine; Bheecarry, Ambicadutt; Zumbo, Betty; Weill, Mylène; Duron, Olivier

    2011-12-01

    The use of the bacterium Wolbachia is an attractive alternative method to control vector populations. In mosquitoes, as in members of the Culex pipiens complex, Wolbachia induces a form of embryonic lethality called cytoplasmic incompatibility, a sperm-egg incompatibility occurring when infected males mate either with uninfected females or with females infected with incompatible Wolbachia strain(s). Here we explore the feasibility of the Incompatible Insect Technique (IIT), a species-specific control approach in which field females are sterilized by inundative releases of incompatible males. We show that the Wolbachia wPip(Is) strain, naturally infecting Cx. p. pipiens mosquitoes from Turkey, is a good candidate to control Cx. p. quinquefasciatus populations on four islands of the south-western Indian Ocean (La Réunion, Mauritius, Grande Glorieuse and Mayotte). The wPip(Is) strain was introduced into the nuclear background of Cx. p. quinquefasciatus mosquitoes from La Réunion, leading to the LR[wPip(Is)] line. Total embryonic lethality was observed in crosses between LR[wPip(Is)] males and all tested field females from the four islands. Interestingly, most crosses involving LR[wPip(Is)] females and field males were also incompatible, which is expected to reduce the impact of any accidental release of LR[wPip(Is)] females. Cage experiments demonstrate that LR[wPip(Is)] males are equally competitive with La Réunion males resulting in demographic crash when LR[wPip(Is)] males were introduced into La Réunion laboratory cages. These results, together with the geographic isolation of the four south-western Indian Ocean islands and their limited land area, support the feasibility of an IIT program using LR[wPip(Is)] males and stimulate the implementation of field tests for a Cx. p. quinquefasciatus control strategy on these islands. PMID:22206033

  10. Development of antimicrobial peptide defenses of southern leopard frogs, Rana sphenocephala, against the pathogenic chytrid fungus, Batrachochytrium dendrobatidis.

    PubMed

    Holden, Whitney M; Reinert, Laura K; Hanlon, Shane M; Parris, Matthew J; Rollins-Smith, Louise A

    2015-01-01

    Amphibian species face the growing threat of extinction due to the emerging fungal pathogen Batrachochytrium dendrobatidis, which causes the disease chytridiomycosis. Antimicrobial peptides (AMPs) produced in granular glands of the skin are an important defense against this pathogen. Little is known about the ontogeny of AMP production or the impact of AMPs on potentially beneficial symbiotic skin bacteria. We show here that Rana (Lithobates) sphenocephala produces a mixture of four AMPs with activity against B. dendrobatidis, and we report the minimum inhibitory concentration (MIC) of synthesized replicates of these four AMPs tested against B. dendrobatidis. Using mass spectrometry and protein quantification assays, we observed that R. sphenocephala does not secrete a mature suite of AMPs until approximately 12 weeks post-metamorphosis, and geographically disparate populations produce a different suite of peptides. Use of norepinephrine to induce maximal secretion significantly reduced levels of culturable skin bacteria. PMID:25218643

  11. Size-sex variation in survival rates and abundance of pig frogs, Rana grylio, in northern Florida wetlands

    USGS Publications Warehouse

    Wood, K.V.; Nichols, J.D.; Percival, H.F.; Hines, J.E.

    1998-01-01

    During 1991-1993, we conducted capture-recapture studies on pig frogs, Rana grylio, in seven study locations in northcentral Florida. Resulting data were used to test hypotheses about variation in survival probability over different size-sex classes of pig frogs. We developed multistate capture-recapture models for the resulting data and used them to estimate survival rates and frog abundance. Tests provided strong evidence of survival differences among size-sex classes, with adult females showing the highest survival probabilities. Adult males and juvenile frogs had lower survival rates that were similar to each other. Adult females were more abundant than adult males in most locations at most sampling occasions. We recommended probabilistic capture-recapture models in general, and multistate models in particular, for robust estimation of demographic parameters in amphibian populations.

  12. Predation efficiency of indigenous larvivorous fish species on Culex pipiens L. larvae (Diptera: Culicidae) in drainage ditches in Argentina.

    PubMed

    Marti, Gerardo A; Azpelicueta, M de las Mercedes; Tranchida, María C; Pelizza, Sebastián A; García, Juan J

    2006-06-01

    Two neotropical freshwater fish species, Cnesterodom decemmaculatus (Poeciliidae) and Jenynsia multidentata (Anablepidae), were collected from human-made ditches, a common habitat of the house mosquito Culex pipiens in La Plata, Argentina. Cnesterodom decemmaculatus was recorded in 62 of the 100 examined ditches, whereas J. multidentata was collected from only 21 ditches sympatrically with C. decemmaculatus. Culex pipiens was the only mosquito species collected, and its larvae and pupae were found in 38 of the 100 ditches. Fish and mosquito larvae and pupae were collected together in only two ditches and were significantly negatively correlated. Siphons of larval Culex and remnants of chironomid larvae, copepods, aquatic mites, and fish were present in the gut contents of two C. decemmaculatus from mosquito-positive ditches, while diatoms and filamentous algae were recorded in every fish dissected. Adult C. decemmaculatus and J. multidentata needed approximately 6.2 h to completely digest one Cx. pipiens 4th instar larva under laboratory conditions. When fish were confined with a density of 60 or fewer Cx. pipiens 4th instar larvae, C. decemmaculatus and J. multidentata adults consumed 100% of them in one day but only 35% and 42%, respectively, when confined with 150 larvae. Eradication of Cx. pipiens from a ditch, where densities had averaged 250 immatures per dip, was achieved 17 days after the introduction of 1,700 C. decemmaculatus. PMID:16859096

  13. Microsatellite characterization of subspecies and their hybrids in Culex pipiens complex mosquitoes along a north-south transect in the central United States of America

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Mosquitoes in the Culex pipiens complex, Cx. p. pipiens L. (Cpp) and Cx. p. quinquefasciatus Say (Cpq), are morphologically similar and important vectors of the West Nile and St. Louis Encephalitis viruses in the US. They hybridize when found in sympatry, which could facilitate the transfer of adva...

  14. Dmrt1 polymorphism covaries with sex-determination patterns in Rana temporaria.

    PubMed

    Ma, Wen-Juan; Rodrigues, Nicolas; Sermier, Roberto; Brelsford, Alan; Perrin, Nicolas

    2016-08-01

    Patterns of sex-chromosome differentiation and gonadal development have been shown to vary among populations of Rana temporaria along a latitudinal transect in Sweden. Frogs from the northern-boreal population of Ammarnäs displayed well-differentiated X and Y haplotypes, early gonadal differentiation, and a perfect match between phenotypic and genotypic sex. In contrast, no differentiated Y haplotypes could be detected in the southern population of Tvedöra, where juveniles furthermore showed delayed gonadal differentiation. Here, we show that Dmrt1, a gene that plays a key role in sex determination and sexual development across all metazoans, displays significant sex differentiation in Tvedöra, with a Y-specific haplotype distinct from Ammarnäs. The differential segment is not only much shorter in Tvedöra than in Ammarnäs, it is also less differentiated and associates with both delayed gonadal differentiation and imperfect match between phenotypic and genotypic sex. Whereas Tvedöra juveniles with a local Y haplotype tend to ultimately develop as males, those without it may nevertheless become functional XX males, but with strongly female-biased progeny. Our findings suggest that the variance in patterns of sex determination documented in common frogs might result from a genetic polymorphism within a small genomic region that contains Dmrt1. They also substantiate the view that recurrent convergences of sex determination toward a limited set of chromosome pairs may result from the co-option of small genomic regions that harbor key genes from the sex-determination pathway. PMID:27551369

  15. Complete sporogony of Plasmodium relictum (lineage pGRW4) in mosquitoes Culex pipiens pipiens, with implications on avian malaria epidemiology.

    PubMed

    Valkiūnas, Gediminas; Žiegytė, Rita; Palinauskas, Vaidas; Bernotienė, Rasa; Bukauskaitė, Dovilė; Ilgūnas, Mikas; Dimitrov, Dimitar; Iezhova, Tatjana A

    2015-08-01

    Plasmodium relictum (lineage pGRW4) causes malaria in birds and is actively transmitted in countries with warm climates and also temperate regions of the New World. In Europe, the lineage pGRW4 has been frequently reported in many species of Afrotropical migrants after their arrival from wintering grounds, but is rare in European resident birds. Obstacles for transmission of this parasite in Europe have not been identified. Culex quinquefasciatus is an effective vector of pGRW4 malaria, but this mosquito is absent from temperate regions of Eurasia. It remains unclear if the lineage pGRW4 completes sporogony in European species of mosquitoes. Here we compare the sporogonic development of P. relictum (pGRW4) in experimentally infected mosquitoes Culex pipiens pipiens form molestus, C. quinquefasciatus, and Ochlerotatus cantans. The pGRW4 parasite was isolated from a garden warbler Sylvia borin, multiplied, and used to infect laboratory-reared Culex spp. and wild-caught Ochlerotatus mosquitoes by allowing them to take blood meals on infected birds. The exposed females were maintained at a mean laboratory temperature of 19 °C, which ranged between 14 °C at night and 24 °C during daytime. They were dissected on intervals to study the development of sporogonic stages. Only ookinetes developed in O. cantans; sporogonic development was abortive. The parasite completed sporogony in both Culex species, with similar patterns of development, and sporozoites were reported in the salivary glands 16 days after infection. The presence of sporogonic stages of the lineage pGRW4 in mosquitoes was confirmed by PCR-based testing of (1) the sporozoites present in salivary glands and (2) the single oocysts, which were obtained by laser microdissection from infected mosquito midguts. This study shows that P. relictum (pGRW4) completes sporogony in C. p. pipiens at relatively low temperatures. We conclude that there are no restrictions for spreading this bird infection in Europe from

  16. Multifarious selection through environmental change: acidity and predator-mediated adaptive divergence in the moor frog (Rana arvalis).

    PubMed

    Egea-Serrano, Andrés; Hangartner, Sandra; Laurila, Anssi; Räsänen, Katja

    2014-04-01

    Environmental change can simultaneously cause abiotic stress and alter biological communities, yet adaptation of natural populations to co-changing environmental factors is poorly understood. We studied adaptation to acid and predator stress in six moor frog (Rana arvalis) populations along an acidification gradient, where abundance of invertebrate predators increases with increasing acidity of R. arvalis breeding ponds. First, we quantified divergence among the populations in anti-predator traits (behaviour and morphology) at different rearing conditions in the laboratory (factorial combinations of acid or neutral pH and the presence or the absence of a caged predator). Second, we evaluated relative fitness (survival) of the populations by exposing tadpoles from the different rearing conditions to predation by free-ranging dragonfly larvae. We found that morphological defences (relative tail depth) as well as survival of tadpoles under predation increased with increasing pond acidity (under most experimental conditions). Tail depth and larval size mediated survival differences among populations, but the contribution of trait divergence to survival was strongly dependent on prior rearing conditions. Our results indicate that R. arvalis populations are adapted to the elevated predator pressure in acidified ponds and emphasize the importance of multifarious selection via both direct (here: pH) and indirect (here: predators) environmental changes. PMID:24552840

  17. Octylphenol induced gene expression in testes of Frog, Rana chensinensis.

    PubMed

    Li, Xinyi; Liu, Jia; Zhang, Yuhui

    2016-06-01

    Octylphenol (OP) is an endocrine-disrupting chemical (EDC), which can disrupt the reproductive system. To understand the effect of OP, a subtractive cDNA library was constructed using suppression subtractive hybridization (SSH) to identify alterations of gene transcription in the testes of the frog Rana chensinensis after OP exposure. Two hundred positive clones were selected and 134 sequences of gene fragments were produced from the subtractive library randomly. These genes were identified to be involved in metabolic process, cellular process, biological regulation, stimulus, immune system and female pregnancy process. In order to verify the efficiency of the subtractive cDNA library, PSG9 and PAPP-A were analyzed further as two representatives of differentially expressed transcription genes using semi-quantitative RT-PCR. Our result was the first successful construction of the subtractive cDNA library in frog testes after OP treatment. Based on this cDNA library, OP was shown to affect multiple physiological processes including inducing immune response, disrupting the steroid hormone synthesis and influencing spermatogenesis in the testis by up-regulation of specific genes. PMID:26896894

  18. Evolutionary dynamics of a rapidly receding southern range boundary in the threatened California red-legged frog (Rana draytonii)

    USGS Publications Warehouse

    Richmond, Jonathan Q.; Barr, Kelly R.; Backlin, Adam R.; Vandergast, Amy G.; Fisher, Robert N.

    2013-01-01

    Populations forming the edge of a species range are often imperiled by isolation and low genetic diversity, with proximity to human population centers being a major determinant of edge stability in modern landscapes. Since the 1960s, the California red-legged frog (Rana draytonii) has undergone extensive declines in heavily urbanized southern California, where the range edge has rapidly contracted northward while shifting its cardinal orientation to an east-west trending axis. We studied the genetic structure and diversity of these frontline populations, tested for signatures of contemporary disturbance, specifically fire, and attempted to disentangle these signals from demographic events extending deeper into the past. Consistent with the genetic expectations of the ‘abundant-center’ model, we found that diversity, admixture, and opportunity for random mating increases in populations sampled successively further away from the range boundary. Demographic simulations indicate that bottlenecks in peripheral isolates are associated with processes extending tens to a few hundred generations in the past, despite the demographic collapse of some due to recent fire-flood events. While the effects of recent disturbance have left little genetic imprint on these populations, they likely contribute to an extinction debt that will lead to continued range contraction unless management intervenes to stall or reverse the process.

  19. Oral chytridiomycosis in the mountain yellow-legged frog (Rana muscosa)

    USGS Publications Warehouse

    Fellers, G.M.; Green, E.D.; Longcore, J.E.

    2001-01-01

    The chytrid fungus Batrachochytrium dendrobatidis was originally reported in wild frog populations in Panama and Australia, and from captive frogs in the U.S. National Zoological Park (Washington, DC). This recently described fungus affects the keratinized epidermis of amphibians and has been implicated as a causative factor in the declines of frog populations. We report here the presence of B. dendrobatidis in larval and recently metamorphosed mountain yellow-legged frogs (Rana muscosa) in or near the Sierra Nevada Mountains of California, an area where declines have been documented in all five species of native anurans. Forty-one percent (158 of 387) of larval R. muscosa examined in the field with a hand lens and 18% (14 of 79) of preserved larvae had abnormalities of the oral disc. Twenty-eight larvae were collected from 10 sites where tadpoles had been observed with missing or abnormally keratinized mouthparts, and 24 of these were examined for infection. Sixty-seven percent (16 of 24) of these tadpoles were infected with B. dendrobatidis. Batrachochytrium dendrobatidis was cultured from both tadpoles and recent metamorphs from one of these sites. Tadpoles with mouthpart abnormalities or confirmed chytrid fungus infections were collected at 23 sites spanning a distance of > 440 km and an elevational range from 1658-3550 m. Life-history traits of R. muscosa may make this species particularly susceptible to infection by Batrachochytrium. We recommend that biologists examine tadpoles for oral disc abnormalities as a preliminary indication of chytridiomycosis. Further, we believe that biologists should take precautions to prevent spreading this and other amphibian diseases from one site to another.

  20. Oral chytridiomycosis in the mountain yellow-legged frog (Rana muscosa)

    USGS Publications Warehouse

    Fellers, G.M.; Green, D.E.; Longcore, J.E.

    2001-01-01

    The chytrid fungus Batrachochytrium dendrobatidis was originally reported in wild frog populations in Panama and Australia, and from captive frogs in the U.S. National Zoological Park (Washington, DC). This recently described fungus affects the keratinized epidermis of amphibians and has been implicated as a causative factor in the declines of frog populations. We report here the presence of B. dendrobatidis in larval and recently metamorphosed mountain yellow-legged frogs (Rana muscosa) in or near the Sierra Nevada Mountains of California, an area where declines have been documented in all five species of native anurans. Forty-one percent (158 of 387) of larval R. muscosa examined in the field with a hand lens and 18% (14 of 79) of preserved larvae had abnormalities of the oral disc. Twenty-eight larvae were collected from 10 sites where tadpoles had been observed with missing or abnormally keratinized mouthparts, and 24 of these were examined for infection. Sixty-seven percent (16 of 24) of these tadpoles were infected with B. dendrobatidis. Batrachochytrium dendrobatidis was cultured from both tadpoles and recent metamorphs from one of these sites. Tadpoles with mouthpart abnormalities or confirmed chytrid fungus infections were collected at 23 sites spanning a distance of > 440 km and an elevational range from 1658a??3550 m. Life-history traits of R. muscosa may make this species particularly susceptible to infection by Batrachochytrium. We recommend that biologists examine tadpoles for oral disc abnormalities as a preliminary indication of chytridiomycosis. Further, we believe that biologists should take precautions to prevent spreading this and other amphibian diseases from one site to another.

  1. Chemical Composition and Larvicidal Activity of Greek Myrtle Essential Oils against Culex pipiens biotype molestus.

    PubMed

    Koutsaviti, Aikaterini; Lignou, Irene; Bazos, Ioannis; Koliopoulos, George; Michaelakis, Antonios; Giatropoulos, Athanassios; Tzakou, Olga

    2015-10-01

    Fresh leaves of Myrtus communis collected from different localities in Greece, were subjected to hydrodistillation and the oils obtained were analyzed by GC-FID and GC-MS. The analyses showed mainly quantitative differences, with the monoterpenes myrtenyl acetate, α-pinene, 1,8-cineole, and linalool, along with limonene, dominating the majority of the analyzed Myrtle oils. The evaluation of the larvicidal activity of the samples against Culex pipiens biotype molestus mosquito showed that all tested samples exhibited moderate to weak toxicity, with cultivated M. communis subsp. communis oil being the most active. PMID:26669120

  2. Essential oils of Satureja species: insecticidal effect on Culex pipiens larvae (Diptera: Culicidae).

    PubMed

    Michaelakis, Antonios; Theotokatos, Spiridon A; Koliopoulos, Georgios; Chorianopoulos, Nikos G

    2007-01-01

    The chemical composition of the essential oils of the wild growing plants of Greek S. spinosa L., S. parnassica subsp. parnassica Heldr.& Sart ex Boiss., S. thymbra and S. montana were determined by GC and GC/MS analysis. The larvicidal activities of the essential oils were assayed against Culex pipiens biotype molestus. The analytical data indicated that various monoterpene hydrocarbons and phenolic monoterpenes constitute the major constituents of the oils, but their concentration varied greatly among the oils examined. The bioassay results indicated that the oils possess significant larvicidal activities and represent an inexpensive source of natural substances mixture that exhibit potentials for use to control the mosquito larvae. PMID:18259143

  3. Heavy Metal Accumulation in Leaves of Hydrocharis Morsus-Ranae L. and Biomonitoring Applications

    NASA Astrophysics Data System (ADS)

    Polechońska, Ludmiła; Dambiec, Małgorzata

    2014-06-01

    In present study the concentrations of Hg, Mn, Zn, Fe and Cu in water, bottom sediments and leaves of Hydrocharis morsus-ranae from 11 oxbow lakes of the Odra River were determined by atomic absorption spectrometry. Trace metal concentration in water and bottom sediments were below the geochemical background, indicating no anthropogenic impact in the studied area. On average, the concentrations of metals in leaves of H. morsus ranae exceeded natural thresholds. A high bioaccumulation factors for metals were recorded. The significant positive correlations found between the content Zn, Fe and Hg of in water and in the H. morsus ranae indicate the potential use of the species in the biomonitoring of environmental contamination with these metals.

  4. Status of the California Red-legged Frog (Rana draytonii) in the State of Baja California, México

    USGS Publications Warehouse

    Peralta-Garcia, Anny; Hellingsworth, Bradford D.; Richmond, Jonathan Q.; Valdez-Villavicencio, Jorge H.; Ruiz-Campos, Gorgonio; Fisher, Robert N.; Cruz-Hernandez, Pedro; Galina-Tessaro, Patricia

    2016-01-01

    The California Red-legged Frog (Rana draytonii) is a threatened species in the United States that has undergone population declines, especially in southern California. Due to the lack of information on the status of Mexican populations, we surveyed for the presence of R. draytonii in Baja California and assessed possible threats to population persistence. Our study area extended from the U.S.-Mexican border to the southern end of the distribution of the species in the Sierra San Pedro Mártir. We found R. draytonii at six of 15 historical sites, none at five proxy sites (i.e., alternative sites chosen because the historical record lacked precise locality data), and four at 24 additional sites. The 10 occupied sites are within three watersheds in the Sierra San Pedro Mártir (two sites at Arroyo San Rafael, two sites at Arroyo San Telmo, and six sites at Arroyo Santo Domingo). We did not detect R. draytonii at 60% of historical sites, including the highest elevation site at La Encantada and multiple low-elevation coastal drainages, suggesting the species has declined in Baja California. The threats we noted most frequently were presence of exotic aquatic animal species, water diversion, and cattle grazing. Management of remaining populations and local education is needed to prevent further declines.

  5. Characterization of the Skin Microbiota in Italian Stream Frogs (Rana italica) Infected and Uninfected by a Cutaneous Parasitic Disease.

    PubMed

    Federici, Ermanno; Rossi, Roberta; Fidati, Laura; Paracucchi, Romina; Scargetta, Silvia; Montalbani, Elena; Franzetti, Andrea; La Porta, Gianandrea; Fagotti, Anna; Simonceli, Francesca; Cenci, Giovanni; Di Rosa, Ines

    2015-01-01

    In human and wildlife populations, the natural microbiota plays an important role in health maintenance and the prevention of emerging infectious diseases. In amphibians, infectious diseases have been closely associated with population decline and extinction worldwide. Skin symbiont communities have been suggested as one of the factors driving the different susceptibilities of amphibians to diseases. The activity of the skin microbiota of amphibians against fungal pathogens, such as Batrachochytrium dendrobatidis, has been examined extensively, whereas its protective role towards the cutaneous infectious diseases caused by Amphibiocystidium parasites has not yet been elucidated in detail. In the present study, we investigated, for the first time, the cutaneous microbiota of the Italian stream frog (Rana italica) and characterized the microbial assemblages of frogs uninfected and infected by Amphibiocystidium using the Illumina next-generation sequencing of 16S rRNA gene fragments. A total of 629 different OTUs belonging to 16 different phyla were detected. Bacterial populations shared by all individuals represented only one fifth of all OTUs and were dominated by a small number of OTUs. Statistical analyses based on Bray-Curtis distances showed that uninfected and infected specimens had distinct cutaneous bacterial community structures. Phylotypes belonging to the genera Janthinobacterium, Pseudomonas, and Flavobacterium were more abundant, and sometimes almost exclusively present, in uninfected than in infected specimens. These bacterial populations, known to exhibit antifungal activity in amphibians, may also play a role in protection against cutaneous infectious diseases caused by Amphibiocystidium parasites. PMID:26370166

  6. Characterization of the Skin Microbiota in Italian Stream Frogs (Rana italica) Infected and Uninfected by a Cutaneous Parasitic Disease

    PubMed Central

    Federici, Ermanno; Rossi, Roberta; Fidati, Laura; Paracucchi, Romina; Scargetta, Silvia; Montalbani, Elena; Franzetti, Andrea; La Porta, Gianandrea; Fagotti, Anna; Simonceli, Francesca; Cenci, Giovanni; Di Rosa, Ines

    2015-01-01

    In human and wildlife populations, the natural microbiota plays an important role in health maintenance and the prevention of emerging infectious diseases. In amphibians, infectious diseases have been closely associated with population decline and extinction worldwide. Skin symbiont communities have been suggested as one of the factors driving the different susceptibilities of amphibians to diseases. The activity of the skin microbiota of amphibians against fungal pathogens, such as Batrachochytrium dendrobatidis, has been examined extensively, whereas its protective role towards the cutaneous infectious diseases caused by Amphibiocystidium parasites has not yet been elucidated in detail. In the present study, we investigated, for the first time, the cutaneous microbiota of the Italian stream frog (Rana italica) and characterized the microbial assemblages of frogs uninfected and infected by Amphibiocystidium using the Illumina next-generation sequencing of 16S rRNA gene fragments. A total of 629 different OTUs belonging to 16 different phyla were detected. Bacterial populations shared by all individuals represented only one fifth of all OTUs and were dominated by a small number of OTUs. Statistical analyses based on Bray-Curtis distances showed that uninfected and infected specimens had distinct cutaneous bacterial community structures. Phylotypes belonging to the genera Janthinobacterium, Pseudomonas, and Flavobacterium were more abundant, and sometimes almost exclusively present, in uninfected than in infected specimens. These bacterial populations, known to exhibit antifungal activity in amphibians, may also play a role in protection against cutaneous infectious diseases caused by Amphibiocystidium parasites. PMID:26370166

  7. The influence of water quality on the embryonic survivorship of the Oregon spotted frog (Rana pretiosa) in British Columbia, Canada.

    PubMed

    McKibbin, René; Dushenko, William T; vanAggelen, Graham; Bishop, Christine A

    2008-05-20

    In Canada, the Oregon spotted frog (Rana pretiosa) is a critically endangered species with only three known populations and an estimated breeding population of less than 400 located in isolated sites in the extreme south-west corner of British Columbia. Floating Nitex cages were used to assess embryonic survivorship in two populations of Oregon spotted frogs from 2002-2005. One population, near Aldergrove, BC experienced declines in population size while the other population, at Maria Slough, increased during the period 1997-2001. During embryo development, we measured trace metals, nutrients and physical parameters in the water at each site. These were used to test the hypothesis that water quality parameters were correlated with embryonic survivorship. During the study period in the declining population at Aldergrove R. pretiosa bred at two distinct sub sites (A and B) located 500 m apart within the wetland. Mean embryonic survivorship varied from 9% to 36% at sub site A and from 78% to 88% at sub site B whereas in the population in Maria Slough, the mean embryonic survivorship varied from 77% to 84%. Sulphate was the only water chemistry variable that differed significantly between the two study sites and was the highest at Maria Slough. A weak significant positive correlation was found between chloride and embryonic survivorship and conductivity and embryonic survivorship. A multiple regression model found conductivity was the only significant variable. We concluded that natural water chemistry conditions of low chloride and consequently low conductivity may be contributing to low embryonic survivorship in the population of R. pretiosa at MD Aldergrove, BC. PMID:18336868

  8. Wolbachia Induces Male-Specific Mortality in the Mosquito Culex pipiens (LIN Strain)

    PubMed Central

    Rasgon, Jason L.

    2012-01-01

    Background Wolbachia are maternally inherited endosymbionts that infect a diverse range of invertebrates, including insects, arachnids, crustaceans and filarial nematodes. Wolbachia are responsible for causing diverse reproductive alterations in their invertebrate hosts that maximize their transmission to the next generation. Evolutionary theory suggests that due to maternal inheritance, Wolbachia should evolve toward mutualism in infected females, but strict maternal inheritance means there is no corresponding force to select for Wolbachia strains that are mutualistic in males. Methodology/Principal findings Using cohort life-table analysis, we demonstrate that in the mosquito Culex pipiens (LIN strain), Wolbachia-infected females show no fitness costs due to infection. However, Wolbachia induces up to a 30% reduction in male lifespan. Conclusions/significance These results indicate that the Wolbachia infection of the Culex pipiens LIN strain is virulent in a sex-specific manner. Under laboratory situations where mosquitoes generally mate at young ages, Wolbachia strains that reduce male survival could evolve by drift because increased mortality in older males is not a significant selective force. PMID:22427798

  9. Efficiency of Colocasia esculenta leaves extract and histopathological effects on Culex pipiens (Diptera: Culicidae).

    PubMed

    El-Monairy, Olfat M

    2015-04-01

    This study evaluated the toxicity of Colocasia esculenta leaves extract on 3rd, 4th instars larvae and pupae of Culex pipiens. Bioassays showed that the 3rd instar larvae was the most susceptible to the different concentrations of extract, where the LC50 after 48 hr. post-exposure was 79.41, 109.65 & 141.25 for the 3rd, 4th instars larvae and pupal stage respectively. The histo-pathological effects of C. esculenta leaves extract on midgut regions and gastric caeca of the 3rd instar larvae were studied. When larvae were treated with 100 ppm of C. esculenta extract, all larvae developed dramatic pathological lesions especially Malpighian tubules were extensively affected. The midgut cells showed morphological deviation from normal ones, through slightly apical degenerated (lysis) of epithelial cells. The epithelial cells with extensive cellular microvilli were shrinkage, the nuclei showed pyknotic characteristic and the peritrophic membrane was appeared discontinuation in compared to control. When the 3rd larval instar was exposed to extract 400 ppm, the epithelial cells, adipose fabric and muscles were extensively affected. Also, the gastric caeca was affected obviously. These observation and alterations in cells of Cx. pipiens larvae are related to the dangerous effect of C. esculent leaves extract. PMID:26012222

  10. Inhibitory Effects of Amorphigenin on the Mitochondrial Complex I of Culex pipiens pallens Coquillett (Diptera: Culicidae)

    PubMed Central

    Ji, Mingshan; Liang, Yaping; Gu, Zumin; Li, Xiuwei

    2015-01-01

    Previous studies in our laboratory found that the extract from seeds of Amorpha fruticosa in the Leguminosae family had lethal effects against mosquito larvae, and an insecticidal compound amorphigenin was isolated. In this study, the inhibitory effects of amorphigenin against the mitochondrial complex I of Culex pipiens pallens (Diptera: Culicidae) were investigated and compared with that of rotenone. The results showed that amorphigenin and rotenone can decrease the mitochondrial complex I activity both in vivo and in vitro as the in vivo IC50 values (the inhibitor concentrations leading to 50% of the enzyme activity lost) were determined to be 2.4329 and 2.5232 μmol/L, respectively, while the in vitro IC50 values were 2.8592 and 3.1375 μmol/L, respectively. Both amorphigenin and rotenone were shown to be reversible and mixed-I type inhibitors of the mitochondrial complex I of Cx. pipiens pallens, indicating that amorphigenin and rotenone inhibited the enzyme activity not only by binding with the free enzyme but also with the enzyme-substrate complex, and the values of KI and KIS for amorphigenin were determined to be 20.58 and 87.55 μM, respectively, while the values for rotenone were 14.04 and 69.23 μM, respectively. PMID:26307964

  11. Repellent activities of essential oils and monoterpenes against Culex pipiens pallens.

    PubMed

    Choi, Won-Sik; Park, Byeoung-Soo; Ku, Sae-Kwang; Lee, Sung-Eun

    2002-12-01

    Essential oils of Eulcalyptus globulus, Lavender officinalis, Rosemarinus officinalis, and Thymus vulgaris were examined for their repellent activities against Culex pipiens pallens. All 4 essential oils effectively repelled adult mosquitoes on hairless mice. Essential oil of T. vulgaris (thyme) had potent repellent activity within the tested materials, with a protection rate of 91% at a concentration of 0.05% topical treatment. Thyme essential oil significantly extended the duration of protection until 3 bites by mosquitoes. With gas chromatography-mass spectrometry analysis, thyme essential oil was a rich source of 5 monoterpenes, including in descending order thymol, p-cymene, carvacrol, linalool, and alpha-terpinene. These 5 monoterpenes also were assessed to determine their repellent activities to the mosquitoes. alpha-Terpinene had a potent repellent activity with a protection rate of 97% at a concentration of 0.05% topical treatment. Additionally, carvacrol and thymol showed an equivalent level of repellency. A spray-type solution containing 2% alpha-terpinene was tested for its repellent activity against Cx. pipiens. This solution showed stronger repellent activity than the currently used repellent N,N-diethyl-m-methylbenzamide (deet). PMID:12542193

  12. Laboratory evaluation of differential attraction of Culex pipiens pallens to fruit-based sugar baits.

    PubMed

    Ding, Yan-Mei; Hu, Yin; Yu, Bao-Ting; Mo, Xiao-Chang; Mo, Jian-Chu

    2016-11-01

    Mosquito adults usually need to obtain sugar from floral nectaries and damaged fruits/seed pods to replenish their energy reserves. The newly developed attractive toxic sugar baits have been successfully applied in controlling various mosquito species outdoors. However, the attraction of Culex pipiens pallens to different fruit-based sugar baits remains unknown. In the present study, we selected nine common fruit species, prepared the fruit-based sugar solutions, and investigated the attractiveness of different sugar baits to newly emerged Cx. pipiens pallens in the laboratory. The results showed that when tested against the 5% brown sugar solution, all the sugar baits were significantly attractive to both females and males. When tested together in the mesh-covered cage, there was a significant difference on the attractiveness between different fruit-based sugar baits. The most attractive fruit species included Broussonetia papyrifera, Cucumis melo, C. melo var. saccharinus, Amygdalus persica and Pyrus bretschneideri, and their seed pods could be potentially used as ingredients in ATSB for controlling mosquitoes outdoors. PMID:27456936

  13. Effects of lead-contaminated sediment on Rana sphenocephala tadpoles

    USGS Publications Warehouse

    Sparling, D.W.; Krest, S.K.; Ortiz-Santaliestra, M.

    2006-01-01

    We exposed larval southern leopard frogs (Rana sphenocephala) to lead-contaminated sediments to determine the lethal and sublethal effects of this metal. Tadpoles were laboratory-raised from early free-swimming stage through metamorphosis at lead concentrations of 45, 75, 180, 540, 2360, 3940, 5520, and 7580 mg/kg dry weight in sediment. Corresponding pore water lead concentrations were 123, 227, 589, 1833, 8121, 13,579, 19,038, and 24,427 ug/L. Tadpoles exposed to lead concentrations in sediment of 3940 mg/kg or higher died within 2 to 5 days of exposure. At lower concentrations, mortality through metamorphosis ranged from 3.5% at 45 mg/kg lead to 37% at 2360 mg/kg lead in sediment. The LC50 value for lead in sediment was 3728 mg/kg (95% CI=1315 to 72,847 mg/kg), which corresponded to 12,539 ug/L lead in pore water (95% CI= 4000 to 35,200 ug/L). Early growth and development were depressed at 2,360 mg/kg lead in sediment (8100 ug/L in pore water) but differences were not evident by the time of metamorphosis. The most obvious effect of lead was its pronounced influence on skeletal development. Whereas tadpoles at 45 mg/kg lead in sediment did not display permanent abnormalities, skeletal malformations increased in frequency and severity at all higher lead concentrations. By 2360 mg/kg, 100% of surviving metamorphs displayed severe spinal problems, reduced femur and humerus lengths, deformed digits, and other bone malformations. Lead concentrations in tissues correlated positively with sediment and pore water concentrations.

  14. Impacts of weathered tire debris on the development of Rana sylvatica larvae.

    PubMed

    Camponelli, Kimberly M; Casey, Ryan E; Snodgrass, Joel W; Lev, Steven M; Landa, Edward R

    2009-02-01

    Highway runoff has the potential to negatively impact receiving systems including stormwater retention ponds where highway particulate matter can accumulate following runoff events. Tire wear particles, which contain about 1% Zn by mass, make up approximately one-third of the vehicle derived particulates in highway runoff and therefore may serve as a stressor to organisms utilizing retention ponds as habitat. In this study, we focused on the potential contribution of tire debris to Zn accumulation by Rana sylvatica larvae and possible lethal or sublethal impacts resulting from exposure to weathered tire debris during development. Eggs and larvae were exposed to aged sediments (containing either ZnCl2 or tire particulate matter, both providing nominal concentrations of 1000 mg Zn kg(-1)) through metamorphosis. Water column Zn was elevated in both the ZnCl2 and tire treatments relative to the control treatment, indicating that aging allowed Zn leaching from tire debris to occur. Tissue Zn was also elevated for the ZnCl2 and tire treatments indicating that Zn in the treatments was available for uptake by the amphibians. Exposure to both ZnCl2 and tire treatments increased the time for larvae to complete metamorphosis in comparison with controls. We also observed that the longer the organisms took to complete metamorphosis, the smaller their mass at metamorphosis. Our results indicate that Zn leached from aged tire debris is bioavailable to developing R. sylvatica larvae and that exposure to tire debris amended sediments can result in measurable physiological outcomes to wood frogs that may influence population dynamics. PMID:18995883

  15. Impacts of weathered tire debris on the development of Rana sylvatica larvae

    USGS Publications Warehouse

    Camponelli, K.M.; Casey, R.E.; Snodgrass, J.W.; Lev, S.M.; Landa, E.R.

    2009-01-01

    Highway runoff has the potential to negatively impact receiving systems including stormwater retention ponds where highway particulate matter can accumulate following runoff events. Tire wear particles, which contain about 1% Zn by mass, make up approximately one-third of the vehicle derived particulates in highway runoff and therefore may serve as a stressor to organisms utilizing retention ponds as habitat. In this study, we focused on the potential contribution of tire debris to Zn accumulation by Rana sylvatica larvae and possible lethal or sublethal impacts resulting from exposure to weathered tire debris during development. Eggs and larvae were exposed to aged sediments (containing either ZnCl2 or tire particulate matter, both providing nominal concentrations of 1000 mg Zn kg-1) through metamorphosis. Water column Zn was elevated in both the ZnCl2 and tire treatments relative to the control treatment, indicating that aging allowed Zn leaching from tire debris to occur. Tissue Zn was also elevated for the ZnCl2 and tire treatments indicating that Zn in the treatments was available for uptake by the amphibians. Exposure to both ZnCl2 and tire treatments increased the time for larvae to complete metamorphosis in comparison with controls. We also observed that the longer the organisms took to complete metamorphosis, the smaller their mass at metamorphosis. Our results indicate that Zn leached from aged tire debris is bioavailable to developing R. sylvatica larvae and that exposure to tire debris amended sediments can result in measurable physiological outcomes to wood frogs that may influence population dynamics. ?? 2008 Elsevier Ltd.

  16. Effects of six chemical deicers on larval wood frogs (Rana sylvatica).

    PubMed

    Harless, Meagan L; Huckins, Casey J; Grant, Jacqualine B; Pypker, Thomas G

    2011-07-01

    Widespread and intensive application of road deicers, primarily road salt (NaCl), in North America threatens water quality and the health of freshwater ecosystems. Intensive use of NaCl can be harmful to sensitive members of freshwater ecosystems such as amphibians. Detection of negative effects of NaCl application has prompted the search for alternative chemical deicers with lower environmental impacts. We conducted a series of 96-h acute toxicity tests to determine the negative sensitivity of larval wood frogs (Rana [Lithobates] sylvatica) to six deicing chemicals: urea (CH(4) N(2) O), sodium chloride (NaCl), magnesium chloride (MgCl(2) ), potassium acetate (CH(3) COOK), calcium chloride (CaCl(2) ), and calcium magnesium acetate (C(8) H(12) CaMgO(8) ). Acetates are sometimes touted as environmentally friendly alternatives to NaCl but have not been examined in enough detail to warrant this designation. When exposed to a range of environmentally realistic concentrations of these chemicals, larvae were least sensitive (i.e., had the lowest mortality rate) to CH(4) N(2) O, NaCl, and MgCl(2) and most sensitive to acetates (C(8) H(12) CaMgO(8) , CH(3) COOK) and CaCl(2) . Our observed median lethal concentration estimates (LC50(96-h) ) for NaCl were over two times higher than values presented in previous studies, which suggests variability in tolerance among R. sylvatica populations. The deicers varied greatly in their toxicity, and further research is warranted to examine the differential effects of this suite of deicers on other species. PMID:21472773

  17. Variation of total mercury concentrations in pig frogs (Rana grylio) across the Florida Everglades, USA.

    PubMed

    Ugarte, Cristina A; Rice, Kenneth G; Donnelly, Maureen A

    2005-06-01

    The Pig Frog (Rana grylio) is an aquatic frog that is an abundant component of the Everglades ecosystem. South Floridians recreationally and commercially hunt pig frogs in marshes throughout Water Conservation Areas (WCA) and Big Cypress National Preserve (BCNP) in South Florida. Most of these areas are under fish consumption advisories because of high levels of methylmercury present in game fish tissues. It is important to understand how mercury is distributed throughout Pig Frog populations because their consumption from certain areas may present a risk to human health. We sampled 88 pig frogs along a north-south transect through the Florida Everglades. There were substantial differences in total mercury (THg) concentrations from leg muscle tissue among sites. Total mercury in frog leg tissue was highest from areas protected from harvest in Everglades National Park (ENP), with a maximum concentration of 2051 ng/g wet mass. The THg levels in R. grylio leg tissue from most harvested areas are below Federal advisory limits. However, many pig frogs collected near Frog City, and one from WCA 3B and 3AN, harvested sites, had THg levels above the USEPA 0.3 mg/kg Fish Tissue Residue Criterion. Spatial patterns in the mercury found among pig frogs were similar to those of other wildlife species from the Everglades. We found frogs to have high THg levels in areas where alligators and mosquito fish also have high THg. THg in ENP frogs had an exponential relationship to SVL, we found no other relationship in frogs from other sites. Our data suggests that pig frogs should not be harvested or consumed from sites that exceed federal limits. PMID:15919527

  18. [Effects of cadmium on metamorphism and gonad differentiation in Rana chensinensis].

    PubMed

    Huang, Min-Yi; Wang, Hong-Yuan; Zhang, Yu-Hui

    2010-06-01

    200 tadpoles of Rana chensinensis at stage 26 - 27 were exposed to 0.05, 0.1, 0.2 or 0.4 mg/L Cd2+ in tap water respectively until they're fully metamorphic after which the heteromorphic young frogs in different treatments were anatomized, females and males were identified through gonad observation, and the female ratio was calculated. Localization of estrogen receptors (ER) in liver cells was investigated in different treatments using immunocytochemistry. The results showed that Cd2+ might induce limb abnormality, however, there was little correlation between abnormality rate and cadmium concentration in lower Cd2+ levels except for a higher limb abnormality ratio in the 0.4 mg/L group. On the other hand, Cd2+ could affect gonad differentiation. Compared to the control group, the proportion of female population increased in the 0.05 mg/L group and decreased in the 0.1, 0.2 and 0.4 mg/L ones. The sex rate in the 0.2 mg/L group is significantly different from that in the control group. Hermaphrodite gonads appeared in the two treatments with 0.2 mg/L and 0.4 mg/L of Cd2+. Additionally, ER expression was positive in both cytoplasm and nucleolus of liver cells in Cd2+ treated groups. But, there was no linear relationship between ER expressions levels and the concentration of Cd2+. These results suggested that cadmium can influence tadpole metamorphosis and gonad development by affecting the secretion of sex hormone. PMID:20672415

  19. Transmission of a pathogenic virus (Iridoviridae) of Culex pipiens larvae mediated by the mermithid Strelkovimermis spiculatus (Nematoda).

    PubMed

    Muttis, Evangelina; Micieli, María Victoria; Urrutia, María Inés; García, Juan José

    2015-07-01

    Little progress been made in elucidating the transmission pathway of the invertebrate iridescent virus (MIV). It has been proposed that the MIV has no active means to enter the mosquito larva. We have previously found that the presence of the mermithid nematode Strelkovimermis spiculatus is associated with MIV infection in Culex pipiens under field conditions. In the present study, we evaluated the transmission of MIV to C. pipiens larvae mediated by S. spiculatus and several factors involved in this pathway (mosquito instars, nematode:mosquito larva ratio, amount of viral inoculum). Our results indicate that S. spiculatus functions as an MIV vector to C. pipiens larvae and seems to be an important pathway of virus entry into this system. Moreover, TEM images of S. spiculatus exposed to the viral suspension showed no infections inside the nematode but showed that viral particles are carried over the cuticle of this mermithid. This explains the correspondence between MIV infection and the factors that affect the parasitism of S. spiculatus in C. pipiens larvae. PMID:26031563

  20. A 3-D QSAR-BASED IDENTIFICATION ALGORITHM FOR POTENTIAL ESTROGEN RECEPTOR LIGANDS

    EPA Science Inventory

    Recent reports concerning the lethal effects of solar ultraviolet-B (UV-B) radiation on amphibians suggest that this stressor has the potential to impact some amphibian populations. In this study embryos and larvae of three anuran species, Rana pipiens, R. clamitans, and R. septe...

  1. Pre-hibernation energy reserves in a temperate anuran, Rana chensinensis, along a relatively fine elevational gradient

    USGS Publications Warehouse

    Lu, X.; Li, B.; Li, Y.; Ma, X.; Fellers, G.M.

    2008-01-01

    Temperate anurans have energy substrates in the liver, fat bodies, carcass and gonads; these stores provide support for metabolism and egg production during hibernation, and for breeding activities in spring. This paper compares the energy budget shortly before hibernation among Rana chensinensis populations at elevations of 1400, 1700 and 2000 m along a river in northern China. The larger frogs, regardless of elevation, had relatively heavy storage organs and the masses of nearly all these organs were positively correlated with each other. After controlling for the effect of body size, we found no significant difference in energetic organ mass among different age classes for each of the three populations. There were sexual differences in energy strategy. Males in all populations accumulated greater reserves in liver, fat bodies and carcass than did females. In contrast, females put more energy into their ovaries and oviducts. Frogs from higher elevations tended to have heavier organs than those from lower elevations; however, the pattern did not vary systematically along fine environmental gradients. Mid-elevation R. chensinensis built up significantly more reserves than low-elevation individuals, but were similar to their highland conspecifics. Males from higher elevations tended to have heavier liver and fat bodies; females were similar in liver and ovary mass across all elevations, but formed heavier fat bodies, oviducts and somatic tissue at higher elevation sites.

  2. Immunostimulatory Activity of Protein Hydrolysate from Oviductus Ranae on Macrophage In Vitro

    PubMed Central

    Huang, Di; Yang, Lubing; Wang, Chenlu; Ma, Sihui; Cui, Li; Huang, Shiyang; Sheng, Xia; Weng, Qiang; Xu, Meiyu

    2014-01-01

    Oviductus Ranae is the dry oviduct of Rana chensinensis, which is also called R. chensinensis oil. Oviductus Ranae is a valuable Chinese crude drug and is recorded in the Pharmacopoeia of the People's Republic of China. The aim of this study was to investigate the immunostimulatory activity of protein hydrolysate of Oviductus Ranae (ORPH) and to assess its possible mechanism. Immunomodulatory activity of ORPH was examined in murine macrophage RAW 264.7 cells. The effect of ORPH on the phagocytic activity of macrophages was determined by the neutral red uptake assay. After treatment with ORPH, NO production levels in the culture supernatant were investigated by Griess assay. The mRNA and protein expressions of inducible nitric oxide synthase (iNOS) were detected by RT-PCR and Western blotting. The production of TNF-α, IL-1β, and IL-6 after treatment with ORPH was measured using ELISA assay. In addition, NF-κB levels were also investigated by Western blot. The results showed that ORPH enhanced the phagocytosis of macrophage, increased productions of TNF-α, IL-1β, IL-6, and NO in RAW 264.7 cells, and upregulated the mRNA and protein expression of iNOS. Besides, NF-κB, levels in RAW 264.7 cells were elevated after ORPH treatment. These findings suggested that ORPH might stimulate macrophage activities by activating the NF-κB pathway. PMID:25610475

  3. Oregon Spotted Frog (Rana pretiosa) movement and demography at Dilman Meadow: implications for future monitoring

    USGS Publications Warehouse

    Chelgren, Nathan D.; Pearl, Christopher A.; Bowerman, Jay; Adams, Michael J.

    2007-01-01

    Introduction The Oregon spotted frog (Rana pretiosa) is a highly aquatic frog that has been extirpated from a large portion of its historic range in the Pacific Northwest, and remaining populations are reduced and isolated (Hayes 1997, Pearl and Hayes 2005). Loss and alteration of marsh habitat, predation and competition from exotic fish and bullfrogs, and degraded water quality from agriculture and livestock grazing are implicated in their decline (Hayes 1997, Pearl and Hayes 2005). In 2001, an interagency team translocated a population of frogs from a site that was to be eliminated by the renovation of the dam impounding Wickiup Reservoir, to newly created ponds at Dilman Meadow (121i?? 39' 52" W, 43i?? 41' 58" N), 2.5 km from the original site in central Oregon, USA. We monitored Oregon spotted frog demography and movements at Dilman Meadow for > 4 yr to assess the efficacy of these mitigation efforts, determine metrics for long-term monitoring, and inform future management at the site. More broadly, many aspects of Oregon spotted frog life history are poorly known, so understanding demography and movement patterns is likely to be useful in its conservation. Although wildlife translocations have been attempted extensively as conservation means, few such projects have been sufficiently monitored for demographic rates to understand the causes for the translocation's success or failure (Dodd and Seigel 1991). Our objective here is to document demographic and movement patterns in the population of Oregon spotted frog at Dilman Meadow so that this information will be available to guide management decisions. To better evaluate amphibian population responses to management actions it is important to consider the contribution of each life history stage and both genders to the balance of reproduction and mortality. Population growth or contraction occurs as a complicated function of the probability of breeding, fecundity, and survival during multiple life history stages

  4. Population.

    ERIC Educational Resources Information Center

    King, Pat; Landahl, John

    This pamphlet has been prepared in response to a new problem, a rapidly increasing population, and a new need, population education. It is designed to help teachers provide their students with some basic population concepts with stress placed on the elements of decision making. In the first section of the pamphlet, some of the basic concepts of…

  5. Species boundaries, phylogeography and conservation genetics of the red-legged frog (Rana aurora/draytonii) complex.

    PubMed

    Shaffer, H Bradley; Fellers, G M; Voss, S Randal; Oliver, J C; Pauly, Gregory B

    2004-09-01

    The red-legged frog, Rana aurora, has been recognized as both a single, polytypic species and as two distinct species since its original description 150 years ago. It is currently recognized as one species with two geographically contiguous subspecies, aurora and draytonii; the latter is protected under the US Endangered Species Act. We present the results of a survey of 50 populations of red-legged frogs from across their range plus four outgroup species for variation in a phylogenetically informative, approximately 400 base pairs (bp) fragment of the mitochondrial cytochrome b gene. Our mtDNA analysis points to several major results. (1) In accord with several other lines of independent evidence, aurora and draytonii are each diagnosably distinct, evolutionary lineages; the mtDNA data indicate that they do not constitute a monophyletic group, but rather that aurora and R. cascadae from the Pacific northwest are sister taxa; (2) the range of the draytonii mtDNA clade extends about 100 km further north in coastal California than was previously suspected, and corresponds closely with the range limits or phylogeographical breaks of several codistributed taxa; (3) a narrow zone of overlap exists in southern Mendocino County between aurora and draytonii haplotypes, rather than a broad intergradation zone; and (4) the critically endangered population of draytonii in Riverside County, CA forms a distinct clade with frogs from Baja California, Mexico. The currently available evidence favours recognition of aurora and draytonii as separate species with a narrow zone of overlap in northern California. PMID:15315679

  6. Pesticides in mountain yellow-legged frogs (Rana muscosa) from the Sierra Nevada Mountains of California, USA

    USGS Publications Warehouse

    Fellers, G.M.; McConnell, L.L.; Pratt, D.; Datta, S.

    2004-01-01

    In 1997, pesticide concentrations were measured in mountain yellow-legged frogs (Rana muscosa) from two areas in the Sierra Nevada Mountains of California, USA. One area (Sixty Lakes Basin, Kings Canyon National Park) had large, apparently healthy populations of frogs. A second area (Tablelands, Sequoia National Park) once had large populations, but the species had been extirpated from this area by the early 1980s. The Tablelands is exposed directly to prevailing winds from agricultural regions to the west. When an experimental reintroduction of R. muscosa in 1994 to 1995 was deemed unsuccessful in 1997, the last 20 (reintroduced) frogs that could be found were collected from the Tablelands, and pesticide concentrations in both frog tissue and the water were measured at both the Tablelands and at reference sites at Sixty Lakes. In frog tissues, dichlorodiphenyldichloroethylene (DDE) concentration was one to two orders of magnitude higher than the other organochlorines (46 ?? 20 ng/g wet wt at Tablelands and 17 ?? 8 Sixty Lakes). Both ??-chlordane and trans-nonachlor were found in significantly greater concentrations in Tablelands frog tissues compared with Sixty Lakes. Organophosphate insecticides, chlorpyrifos, and diazinon were observed primarily in surface water with higher concentrations at the Tablelands sites. No contaminants were significantly higher in our Sixty Lakes samples.

  7. Radiation-induced micronuclei in peripheral erythrocytes of Rana catesbeiana: an aquatic animal model for in vivo genotoxicity studies

    SciTech Connect

    Krauter, P.W.; Anderson, S.L.; Harrison, F.L.

    1987-01-01

    An in vivo micronucleus assay for peripheral erythrocytes of Rana catesbeiana tadpoles was developed and evaluated. The assay was used to determine the spontaneous frequency of micronuclei in circulating erythrocytes in tadpoles from two different populations, to define the time from administering the clastogen to the maximum micronucleus frequency in peripheral erythrocytes, and to determine the response to radiation. The spontaneous frequency of micronuclei in circulating erythrocytes of early-stage tadpoles was low, but higher than that of late-stage tadpoles. The time from the exposure of early-stage tadpoles to radiation (2.1 Gy) to the maximum micronucleus frequency was about 2 wk. The increase in frequency of micronuclei in peripheral erythrocytes of late-stage tadpoles receiving doses ranging from 0.5 to 3.0 Gy was linear with dose; a 3-fold increase was obtained with a dose of 3.0 Gy. The spontaneous frequency of micronuclei in erythrocytes and the increase in frequency induced by radiation appeared to differ in tadpoles from different populations. Quantification of micronuclei in the peripheral erythrocytes of R castesbeiana tadpoles provides a promising whole-animal system for studies of genotoxicity in aquatic environments.

  8. Radiation-induced micronuclei in peripheral erythrocytes of Rana catesbeiana: an aquatic animal model for in vivo genotoxicity studies

    SciTech Connect

    Krauter, P.W.; Anderson, S.L.; Harrison, F.L.

    1987-01-01

    An in vivo micronucleus assay for peripheral erythrocytes of Rana catesbeiana tadpoles was developed and evaluated. The assay was used to determine the spontaneous frequency of micronuclei in circulating erythrocytes in tadpoles from two different populations, to define the time from administering the clastogen to the maximum micronucleus frequency in peripheral erythrocytes, and to determine the response to radiation. The spontaneous frequency of micronuclei in circulating erythrocytes of early-stage tadpoles was low (3.6 +/- 2.8 micronuclei per 1,000 erythrocytes, MN o/oo), but higher than that of late-stage tadpoles (1.7 +/- 0.7 MN o/oo). The time from the exposure of early-stage tadpoles to radiation (2.1 Gy) to the maximum micronucleus frequency was about 2 wk. The increase in frequency of micronuclei in peripheral erythrocytes of late-stage tadpoles receiving doses ranging from 0.5 to 3.0 Gy was linear with dose; a 3-fold increase was obtained with a dose of 3.0 Gy. The spontaneous frequency of micronuclei in erythrocytes and the increase in frequency induced by radiation appeared to differ in tadpoles from different populations. Quantification of micronuclei in the peripheral erythrocytes of R catesbeiana tadpoles provides a promising whole-animal system for studies of genotoxicity in aquatic environments.

  9. Insecticidal properties of essential plant oils against the mosquito Culex pipiens molestus (Diptera: Culicidae).

    PubMed

    Traboulsi, Abdallah F; Taoubi, K; el-Haj, Samih; Bessiere, J M; Rammal, Salma

    2002-05-01

    The insecticidal activities of essential oil extracts from leaves and flowers of aromatic plants against fourth-instar larvae of the mosquito Culex pipiens molestus Forskal were determined. Extracts of Myrtus communis L were found to be the most toxic, followed by those of Origanum syriacum L, Mentha microcorphylla Koch, Pistacia lentiscus L and Lavandula stoechas L with LC50 values of 16, 36, 39, 70 and 89 mg litre-1, respectively. Over 20 major components were identified in extracts from each plant species. Eight pure components (1,8-cineole, menthone, linalool, terpineol, carvacrol, thymol, (1S)-(-)-alpha-pinene and (1R)-(+)-alpha-pinene) were tested against the larvae. Thymol, carvacrol, (1R)-(+)-alpha-pinene and (1S)-(-)-alpha-pinene were the most toxic (LC50 = 36-49 mg litre-1), while menthone, 1,8-cineole, linalool and terpineol (LC50 = 156-194 mg litre-1) were less toxic. PMID:11997977

  10. The significance of the Culex pipiens fatigans Wiedemann problem in Ceylon*

    PubMed Central

    Abdulcader, M. H. M.

    1967-01-01

    Bancroftian filariasis became a problem in Ceylon after the Second World War. The only vector is Culex pipiens fatigans, which is widely distributed throughout the country and is prevalent throughout the year. It is a domestic mosquito found resting mainly on hangings and household articles. Its biting hours are from dusk to dawn, with peaks around midnight and in the early hours of the morning. So far this mosquito has not been incriminated as either the natural or the laboratory vector of animal filariasis in Ceylon. C. p. fatigans collected from northern, eastern and north-central parts of the country are capable of transmitting bancroftian infection. This mosquito was able to survive after ingesting high numbers of microfilariae. The main larval habitat consists of collections of polluted water. Husk pits, underground drains and receptables around houses constitute special problems. The control results so far obtained are encouraging. PMID:5300061

  11. Use of femur bone density to segregate wild from farmed Dybowski's frog (Rana dybowskii).

    PubMed

    Yang, Shu Hui; Huang, Xiao Ming; Xia, Rui; Xu, Yan Chun; Dahmer, Thomas D

    2011-04-15

    Wildlife has been utilized by humans throughout history and demand continues to grow today. Farming of wildlife can supplement the supply of wild-harvested wildlife products and, in theory, can reduce pressure on free-ranging populations. However, poached wildlife products frequently enter legal markets where they are fraudulently sold as farmed wildlife products. To effectively close this illegal trade in wild-captured wildlife, there is a need to discriminate wild products from farmed products. Because of the strong market demand for wild-captured frog meat and the resulting strong downward pressure on wild populations, we undertook research to develop a method to discriminate wild from farmed Dybowski's frog (Rana dybowskii) based on femur bone density. We measured femur bone density (D(f)) as the ratio of bone mass to bone volume. D(f) of wild frogs revealed a slightly increasing linear trend with increasing age (R(2)=0.214 in males and R(2)=0.111 in females, p=0.000). Wild males and wild females of age classes from 2 to ≥ 5 years had similar D(f) values. In contrast, 2-year-old farmed frogs showed significantly higher D(f) values (p=0.000) among males (mean D(f)=0.623 ± 0.011 g/ml, n=32) than females (mean D(f)=0.558 ± 0.011 g/ml, n=27). For both sexes, D(f) of wild frogs was significantly higher than that of farmed frogs (p=0.000). Among males, 87.5% (28 of 32 individuals) of farmed frogs were correctly identified as farmed frogs and 86.3% (69 of 80 individuals) of wild frogs were correctly identified as wild frogs. These results suggest that femur bone density is one reliable tool for discriminating between wild and farmed Dybowski's frog. This study also highlights a novel strategy with explicit forensic potential to discriminate wild from captive bred wildlife species. PMID:20933347

  12. Spatiotemporal Diversification of the True Frogs (Genus Rana): A Historical Framework for a Widely Studied Group of Model Organisms.

    PubMed

    Yuan, Zhi-Yong; Zhou, Wei-Wei; Chen, Xin; Poyarkov, Nikolay A; Chen, Hong-Man; Jang-Liaw, Nian-Hong; Chou, Wen-Hao; Matzke, Nicholas J; Iizuka, Koji; Min, Mi-Sook; Kuzmin, Sergius L; Zhang, Ya-Ping; Cannatella, David C; Hillis, David M; Che, Jing

    2016-09-01

    True frogs of the genus Rana are widely used as model organisms in studies of development, genetics, physiology, ecology, behavior, and evolution. Comparative studies among the more than 100 species of Rana rely on an understanding of the evolutionary history and patterns of diversification of the group. We estimate a well-resolved, time-calibrated phylogeny from sequences of six nuclear and three mitochondrial loci sampled from most species of Rana, and use that phylogeny to clarify the group's diversification and global biogeography. Our analyses consistently support an "Out of Asia" pattern with two independent dispersals of Rana from East Asia to North America via Beringian land bridges. The more species-rich lineage of New World Rana appears to have experienced a rapid radiation following its colonization of the New World, especially with its expansion into montane and tropical areas of Mexico, Central America, and South America. In contrast, Old World Rana exhibit different trajectories of diversification; diversification in the Old World began very slowly and later underwent a distinct increase in speciation rate around 29-18 Ma. Net diversification is associated with environmental changes and especially intensive tectonic movements along the Asian margin from the Oligocene to early Miocene. Our phylogeny further suggests that previous classifications were misled by morphological homoplasy and plesiomorphic color patterns, as well as a reliance primarily on mitochondrial genes. We provide a phylogenetic taxonomy based on analyses of multiple nuclear and mitochondrial gene loci. [Amphibians; biogeography; diversification rate; Holarctic; transcontinental dispersal. PMID:27288482

  13. 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. PMID:25983113

  14. Pesticides and Population Declines of California Alpine Frogs

    EPA Science Inventory

    Airborne pesticides from the Central Valley of California have been implicated as a cause for population declines of several amphibian species, with the strongest evidence for the mountain yellow-legged frog complex (Rana muscosa and R. sierrae) in the Sierra Nevada. We measured ...

  15. Trapping biases of Culex torrentium and Culex pipiens revealed by comparison of captures in CDC traps, ovitraps, and gravid traps.

    PubMed

    Hesson, Jenny C; Ignell, Rickard; Hill, Sharon R; Östman, Örjan; Lundström, Jan O

    2015-06-01

    We evaluate three trapping methods for their effectiveness at capturing Culex pipiens and Culex torrentium, both enzootic vectors of bird-associated viruses in Europe. The comparisons, performed in two regions in Sweden, were among CDC traps baited with carbon dioxide, gravid traps, and ovitraps baited with hay infusion. The proportions of the two Culex species in a catch differed between trap types, with CDC traps catching a lower proportion of Cx. torrentium than both gravid traps and ovitraps. Between gravid traps and ovitraps, there was no difference in the proportions of the two species. The results indicate that Cx. torrentium may go undetected or underestimated compared to Cx. pipiens when using carbon dioxide baited CDC traps. The new insight of trap bias presented here adds an important dimension to consider when investigating these vectors of bird-associated viruses in the field. PMID:26047196

  16. Detection of West Nile viral RNA from an overwintering pool of Culex pipens pipiens (Diptera: Culicidae) in New Jersey, 2003.

    PubMed

    Farajollahi, Ary; Crans, Wayne J; Bryant, Patricia; Wolf, Bruce; Burkhalter, Kristen L; Godsey, Marvin S; Aspen, Stephen E; Nasci, Roger S

    2005-05-01

    In total, 1,324 Culex pipiens pipiens L. female mosquitoes were collected at Ft. Hancock, Monmouth County, New Jersey, from January to March 2001-2003. Mosquitoes were held in an insectary at 27 degrees C and a photoperiod of 16:8 (L:D) h for 6 to 21 d after which they were tested in 34 pools. West Nile viral RNA was detected in one pool by a TaqMan reverse transcription-polymerase chain reaction assay; however, infectious virus could not be isolated using either Vero cell plaque assay or C6/36 mosquito cells. Twenty females dissected in January and March 2003 confirmed ovarian diapause status. We suggest that the mode of infection in this pool of overwintering females may have been due to vertical (transgenerational) transmission. PMID:15962803

  17. Experimental Repatriation of Mountain Yellow-legged Frogs (Rana muscosa) in the Sierra Nevada of California

    USGS Publications Warehouse

    Fellers, Gary M.; Bradford, David F.; Pratt, David; Wood, Leslie

    2008-01-01

    In the late 1970s, Rana muscosa (mountain yellow-legged frog) was common in the Tableland area of Sequoia National Park, California where it was possible to find hundreds of tadpoles and adults around many of the ponds and lakes. Surveys in 1993-1995 demonstrated that R. muscosa was absent from more than half of all suitable habitat within the park, including the Tableland area. At that same time, R. muscosa was still common at Sixty Lake Basin, Kings Canyon National Park, 30 km to the northeast. To evaluate the potential causes for the extirpation, we repatriated R. muscosa eggs, tadpoles, subadults, and adult frogs from Sixty Lake Basin to four sites in the Tableland area in 1994 and 1995. We subsequently surveyed each release site and the surrounding area 2 - 3 times per week in 1994-1995, and intermittently in 1996-1997, to monitor the survival of all life history stages, and to detect dispersal of adults and subadults. We also monitored predation, water quality, weather, and water temperature. Our techniques for capturing, holding, transporting, and releasing R. muscosa were refined during the study, and during 1995 resulted in high initial survival rates of all life history stages. Adult frogs were anaesthetized, weighed, measured, tagged, and held in plastic boxes with wet paper towels. Tadpoles were collected and held in fiberglass screen cages set in the water at the edge of a pond. This resulted in relatively natural conditions with less crowding and good water circulation. Frogs, tadpoles, and eggs were placed in Ziploc bags for transport to the Tableland by helicopter. Short-term survival of tadpoles, subadults, and adults was high at all four release sites, tadpoles reached metamorphosis, and adult frogs were still present. However, we detected no evidence of reproduction at three sites (e.g., no new eggs or small tadpoles) and nearly all life history stages disappeared within 12 months. At the fourth site, there was limited reproduction, but it was

  18. Potential of biologically active plant oils to control mosquito larvae (Culex pipiens, Diptera: Culicidae) from an Egyptian locality.

    PubMed

    Khater, Hanem Fathy; Shalaby, Afaf Abdel-Salam

    2008-01-01

    The insecticidal effect of six commercially available plant oils was tested against 4th larval instars of Culex pipiens. Larvae were originally collected from Meit El-Attar, Qalyubia Governorate, Egypt, and then reared in the laboratory until F1 generation. The LC50 values were 32.42, 47.17, 71.37, 83.36, 86.06, and 152.94 ppm for fenugreek (Trigonella foenum-grecum), earth almond (Cyperus esculentus), mustard (Brassica compestris), olibanum (Boswellia serrata), rocket (Eruca sativa), and parsley (Carum ptroselinum), respectively. The tested oils altered some biological aspects of C. pipiens, for instance, developmental periods, pupation rates, and adult emergences. The lowest concentrations of olibanum and fenugreek oils caused remarkable prolongation of larval and pupal durations. Data also showed that the increase of concentrations was directly proportional to reduction in pupation rates and adult emergences. Remarkable decrease in pupation rate was achieved by mustard oil at 1000 ppm. Adult emergence was suppressed by earth almond and fenugreek oils at 25 ppm. In addition, the tested plant oils exhibited various morphological abnormalities on larvae, pupae, and adult stages. Consequently, fenugreek was the most potent oil and the major cause of malformation of both larval and pupal stages. Potency of the applied plant oils provided an excellent potential for controlling C. pipiens. PMID:18488090

  19. Survivorship and fecundity of Culex pipiens pallens feeding on flowering plants and seed pods with differential preferences.

    PubMed

    Yu, Bao-Ting; Ding, Yan-Mei; Mo, Xiao-Chang; Liu, Ning; Li, Hong-Jie; Mo, Jian-Chu

    2016-03-01

    Adult mosquitoes rely on ingestion of sugar from plants to survive, swarm and mate. Culex pipiens pallens Coguillett is the primary vector of lymphatic filariasis and epidemic encephalitis. Little is known about the effect of feeding on different sugar sources on the survivorship and fecundity of Cx. pipiens pallens. In the present study, newly emerged mosquitoes were exposed to several flowering plant and seed pod species with different olfactory preferences, and the survival times of mosquitoes exposed to these sugar sources were determined. The proportions of mosquitoes that ingested sugar from host plants were investigated by cold anthrone tests. The numbers of eggs per egg raft laid by mosquitoes were compared when they were provided with different sugar sources and one blood meal. The results revealed that feeding on different kinds of sugar sources significantly affected female and male mosquitoes' survival times. Cold anthrone tests indicated that the proportions of sugar-positive mosquitoes from different nutritional regimes within 24h corresponded to the preference rankings of Cx. pipiens pallens to these sugar sources, and rapid declines in the proportions of surviving individuals might be attributed to their insufficient ingestion of sugar from nutritional regimes. Feeding on different sugar sources strongly affected the proportions of engorged mosquitoes, and females that had fed on their preferred sugar sources laid more eggs than mosquitoes provided with less preferred sugar sources. The results would provide insights in developing mosquito control strategies that target the sugar feeding behavior of mosquitoes. PMID:26739652

  20. Spatial Variation in Host Feeding Patterns of Culex tarsalis and the Culex pipiens complex (Diptera: Culicidae) in California

    PubMed Central

    THIEMANN, T. C.; LEMENAGER, D. A.; KLUH, S.; CARROLL, B. D.; LOTHROP, H. D.; REISEN, W. K.

    2012-01-01

    West Nile virus (family Flaviviridae, genus Flavivirus, WNV) is now endemic in California across a variety of ecological regions that support a wide diversity of potential avian and mammalian host species. Because different avian hosts have varying competence for WNV, determining the blood-feeding patterns of Culex (Diptera: Culicidae) vectors is a key component in understanding the maintenance and amplification of the virus as well as tangential transmission to humans and horses. We investigated the blood-feeding patterns of Culex tarsalis Coquillett and members of the Culex pipiens L. complex from southern to northern California. Nearly 100 different host species were identified from 1,487 bloodmeals, by using the mitochondrial gene cytochrome c oxidase I (COI). Cx. tarsalis fed on a higher diversity of hosts and more frequently on nonhuman mammals than did the Cx. pipiens complex. Several WNV-competent host species, including house finch and house sparrow, were common bloodmeal sources for both vector species across several biomes and could account for WNV maintenance and amplification in these areas. Highly competent American crow, western scrub-jay and yellow-billed magpie also were fed upon often when available and are likely important as amplifying hosts for WNV in some areas. Neither species fed frequently on humans (Cx. pipiens complex [0.4%], Cx. tarsalis [0.2%]), but with high abundance, both species could serve as both enzootic and bridge vectors for WNV. PMID:22897051

  1. Flight capacity of adult Culex pipiens pallens (Diptera: Culicidae) in relation to gender and day-age.

    PubMed

    Cui, Jianxin; Li, Shujuan; Zhao, Ping; Zou, Fumin

    2013-09-01

    Culex pipiens pallens (L.) is the most common mosquito in houses of central and northern China. It is the primary vector of lymphatic filariasis and Japanese encephalitis. The flight range of mosquitoes is an important factor predicting the risk area of transmission of mosquito-borne pathogens to vertebrate hosts. The flight performance of Cx. pipiens pallens was measured with a 26-channel computer-monitored flight-mill system. We found that females had longer flight capability than males for total flight distance (TFD) and total flight duration (TFDr), and females flew faster than males based on mean flight velocity. No significant difference in flight capability was found between different age-groups in males. However, certain age-groups of females showed significant differences in TFDr and TFD. Specifically, TFD and TFDr tended to be shortest for 5- and 6-d-old females. These significant differences in flight capability between ages and genders provide insights to determine the size of operational area to achieve effective control of Cx. pipiens pallens and minimize the risk of the related mosquito-borne epidemic diseases of lymphatic filariasis and Japanese encephalitis. PMID:24180110

  2. Growth and developmental effects of coal combustion residues on Southern Leopard Frog (Rana sphenocephala) tadpoles exposed throughout metamorphosis

    SciTech Connect

    Peterson, J.D.; Peterson, V.A.; Mendonca, M.T.

    2008-09-15

    The effects of aquatic deposition of coal combustion residues (CCRs) on amphibian life histories have been the focus of many recent studies. In summer 2005, we raised larval Southern Leopard Frogs, Rana sphenocephala, on either sand or CCR substrate (approximately 1 cm deep within plastic bins) and documented effects of sediment type on oral disc condition, as well as time to, mass at, and total body length at key developmental stages, including metamorphosis (Gosner stages (GS) 37, 42, and 46). We found no significant difference in mortality between the two treatments and mortality was relatively low (eight of 48 in the control group and four of 48 in the CCR group). Ninety percent of exposed tadpoles displayed oral disc abnormalities, while no control individuals displayed abnormalities. Tadpoles raised on CCR-contaminated sediment had decreased developmental rates and weighed significantly less at all developmental stages, on average, when compared to controls. The CCR treatment group was also significantly shorter In length than controls at the completion of metamorphosis (GS 46). Collectively, these findings are the most severe sub-lethal effects noted for any amphibian exposed to CCRs to date. More research is needed to understand how these long term effects may contribute to the dynamics of local amphibian populations.

  3. Local adaptation with high gene flow: temperature parameters drive adaptation to altitude in the common frog (Rana temporaria)

    PubMed Central

    Muir, A P; Biek, R; Thomas, R; Mable, B K

    2014-01-01

    Both environmental and genetic influences can result in phenotypic variation. Quantifying the relative contributions of local adaptation and phenotypic plasticity to phenotypes is key to understanding the effect of environmental variation on populations. Identifying the selective pressures that drive divergence is an important, but often lacking, next step. High gene flow between high- and low-altitude common frog (Rana temporaria) breeding sites has previously been demonstrated in Scotland. The aim of this study was to assess whether local adaptation occurs in the face of high gene flow and to identify potential environmental selection pressures that drive adaptation. Phenotypic variation in larval traits was quantified in R. temporaria from paired high- and low-altitude sites using three common temperature treatments. Local adaptation was assessed using QST–FST analyses, and quantitative phenotypic divergence was related to environmental parameters using Mantel tests. Although evidence of local adaptation was found for all traits measured, only variation in larval period and growth rate was consistent with adaptation to altitude. Moreover, this was only evident in the three mountains with the highest high-altitude sites. This variation was correlated with mean summer and winter temperatures, suggesting that temperature parameters are potentially strong selective pressures maintaining local adaptation, despite high gene flow. PMID:24330274

  4. Heavy metals alter the survival, growth, metamorphosis, and antipredatory behavior of Columbia spotted frog (Rana luteiventris) tadpoles.

    PubMed

    Lefcort, H; Meguire, R A; Wilson, L H; Ettinger, W F

    1998-10-01

    Amphibian populations appear to be declining around the world. Although there is no single cause, one factor may be pollution from heavy metals. As a result of mining in the Silver Valley of Idaho, heavy metals have been released into habitats containing many species of sensitive organisms, including spotted frogs (Rana luteiventris). While the gross extent of pollution has been well documented, the more subtle behavioral effects of heavy metals such as lead, zinc, and cadmium are less well studied. We tested the effects of heavy metals on the short-term survival (LC50) of spotted frog tadpoles. Compared to single metals, metals presented together were toxic at lower doses. We also raised the tadpoles in outdoor mini-ecosystems containing either a single heavy metal or soil from an EPA Superfund site in the Silver Valley known to be composed of numerous heavy metals. Exposure to Silver Valley soil resulted in delayed metamorphosis. We tested the ability of metal-exposed tadpoles to detect and respond to chemical cues emanating from predacious rainbow trout. We found that high levels of Silver Valley soil, medium levels of zinc, and medium and high levels of lead resulted in a decreased fright response. Low levels of cadmium, zinc, and lead did not cause a significant effect, but low levels of soil did result in a decreased fright response. Heavy metals may alter interactions between tadpoles and their predators. PMID:9732476

  5. Long-term effects of pesticide exposure at various life stages of the southern leopard frog (Rana sphenocephala)

    USGS Publications Warehouse

    Bridges, C.M.

    2000-01-01

    Amphibian larvae are commonly exposed to low levels of pesticides during their development. Chronic studies generally examine the effects of long-term exposure, but they often disregard the importance of the individual life stage at which tadpoles are exposed. I determined the point during development at which carbaryl effects are manifested by exposing southern leopard frog tadpoles (Rana sphenocephala) to the pesticide carbaryl at five different times during development. Metamorphs exposed throughout the tadpole stage and throughout development (egg, embryo, tadpole) experienced significant mortality at all chemical levels. Although the length of the larval period was the same for all experimental groups, metamorphs exposed during the egg stage were smaller than their corresponding controls, independent of whether they were exposed at any other stage. Nearly 18% of individuals exposed to carbaryl during development exhibited some type of developmental deformity (including both visceral and limb malformities), compared to a single deformed (< 1%) control tadpole, demonstrating that a chemical hypothesis for amphibian deformities remains viable. Because exposure to nonpersistent chemicals may last for only a short period of time, it is important to examine the long-term effects that short-term exposure has on larval amphibians and the existence of any sensitive life stage. Any delay in metamorphosis or decrease in size at metamorphosis can impact demographic processes of the population, potentially leading to declines or local extinction.

  6. Biospectroscopy reveals the effect of varying water quality on tadpole tissues of the common frog (Rana temporaria).

    PubMed

    Strong, Rebecca J; Halsall, Crispin J; Ferenčík, Martin; Jones, Kevin C; Shore, Richard F; Martin, Francis L

    2016-06-01

    Amphibians are undergoing large population declines in many regions around the world. As environmental pollution from both agricultural and urban sources has been implicated in such declines, there is a need for a biomonitoring approach to study potential impacts on this vulnerable class of organism. This study assessed the use of infrared (IR) spectroscopy as a tool to detect changes in several tissues (liver, muscle, kidney, heart and skin) of late-stage common frog (Rana temporaria) tadpoles collected from ponds with differing water quality. Small differences in spectral signatures were revealed between a rural agricultural pond and an urban pond receiving wastewater and landfill run-off; these were limited to the liver and heart, although large differences in body size were apparent, surprisingly with tadpoles from the urban site larger than those from the rural site. Large differences in liver spectra were found between tadpoles from the pesticide and nutrient impacted pond compared to the rural agricultural pond, particularly in regions associated with lipids. Liver mass and hepatosomatic indices were found to be significantly increased in tadpoles from the site impacted by pesticides and trace organic chemicals, suggestive of exposure to environmental contamination. Significant alterations were also found in muscle tissue between tadpoles from these two ponds in regions associated with glycogen, potentially indicative of a stress response. This study highlights the use of IR spectroscopy, a low-cost, rapid and reagent-free technique in the biomonitoring of a class of organisms susceptible to environmental degradation. PMID:26925755

  7. Time of arrival of gravid Culex pipiens fatigans at an oviposition site, the oviposition cycle and the relationship between time of feeding and time of oviposition.

    PubMed

    de Meillon, B; Sebastian, A; Khan, Z H

    1967-01-01

    One of the most important activities in a female mosquito's life is the flight to the breeding place and the subsequent deposition of eggs. During this phase, motivated by endogenous and exogenous stimuli, the female is particularly exposed and susceptible to attack. It is therefore important to investigate these episodes in the gravid female's life.The work reported in this paper shows that gravid Culex pipiens fatigans females are easily trapped over breeding-water; there are two peaks in the arrival at a breeding site, one just after sunset and the other at sunrise. The oviposition cycle is biphasic, the two peaks coinciding, in calm weather, with the two arrival peaks; wind and rain cause marked disturbances in the oviposition cycle.The mean duration of the gonotrophic cycle depends on the time of feeding; this finding is of practical importance since the length of the cycle is often used to calculate the daily survival rate of adult mosquitos. It appears that oviposition is stimulated by a change in light: from light to dark for mosquitos ovipositing in the evening and from dark to light for those ovipositing in the morning.Apart from revealing some hitherto unknown behaviour patterns, the techniques evolved could also be used in the assessment of mosquito populations and hence the effects of control measures. PMID:4227196

  8. Investigation of the anti-glioma activity of Oviductus ranae protein hydrolysate.

    PubMed

    Sui, Xin; Li, Xiao-Hua; Duan, Ming-Hua; Jia, Ai-Ling; Wang, Ye; Liu, Da; Li, Yi-Ping; Qiu, Zhi-Dong

    2016-07-01

    Oviductus Ranae is the dry oviducts of Rana temporaria chensinensis, and it has been reported to have a range of biological activities. This study aimed to investigate the effects of Oviductus Ranae protein hydrolysate (ORPH) on human glioma C6 cell proliferation and apoptosis in vitro and in vivo. Following in vitro treatment, cell viability and colony formation assays showed that ORPH inhibited C6 cell proliferation. In addition, the results of western blotting also demonstrated that ORPH effectively regulated the expression of the apoptosis related proteins, cleaved caspase-3, Bax and Bcl-2, DNA staining and flow cytometry analysis demonstrated that ORPH significantly promoted apoptosis in this cell line, a finding that was confirmed in vivo using terminal deoxynucleotidyl transferase dUTP nick end labeling. Further investigation demonstrated that ORPH increased apoptosis by modulating the release of inflammatory cytokines and the phosphoinositide 3-kinase (PI3K)/AKT signaling pathway; this was demonstrated using a PI3K/AKT inhibitor (NVP-BEZ235). In summary, the present study suggested that ORPH promoted apoptosis and inhibited glioma cell proliferation by influencing the PI3K/AKT signaling pathway. PMID:27261592

  9. Asymmetrical Effects of Introduced Bullfrogs (Rana catesbeiana) on Native Ranid Frogs in Oregon

    USGS Publications Warehouse

    Pearl, C.A.; Adams, M.J.; Bury, R.B.; McCreary, B.

    2004-01-01

    Introduced American Bullfrogs (Rana catesbeiana) have become widely established in the Pacific Northwest over the last century and are thought to be an important predator of native amphibians throughout the western United States. The Northern Red-Legged Frog (Rana aurora aurora) and Oregon Spotted Frog (Rana pretiosa) historically coexisted in portions of the Pacific Northwest now invaded by R. catesbeiana, but R. pretiosa has declined more severely than R. a. aurora. We investigated whether microhabitat and behavioral differences that facilitate sympatric coexistence of the natives predict which species is more susceptible to predation by introduced R. catesbeiana. Our laboratory experiments demonstrate that R. catesbeiana adults prefer aquatic microhabitats, that R. pretiosa juveniles are more aquatic than R. a. aurora, and that adult R. catesbeiana consume more R. pretiosa than R. a. aurora juveniles. Mean and maximum jump distances of R. pretiosa were shorter than equally sized R. a. aurora, and the difference between these two species increased with larger frog sizes. Our examination of field survey data indicates that R. pretiosa coexist with R. catesbeiana less frequently than R. a. aurora. We conclude that R. catesbeiana is a greater threat to survival of R. pretiosa than to R. a. aurora and suggest that microhabitat use and escape abilities of native ranid frogs may be linked to this asymmetrical effect. Analysis of behavioral and microhabitat differences among related native species may be a useful tool in predicting the effects of introduced predators on amphibians and can assist in developing conservation priorities for these species.

  10. Asymmetrical effects of introduced Rana catesbeiana on native ranid frogs in Oregon, USA

    USGS Publications Warehouse

    Pearl, Christopher A.; Adams, Michael J.; Bury, R. Bruce; McCreary, B.

    2004-01-01

    Introduced American Bullfrogs (Rana catesbeiana) have become widely established in the Pacific Northwest over the last century and are thought to be an important predator of native amphibians throughout the western United States. The Northern Red-Legged Frog (Rana aurora aurora) and Oregon Spotted Frog (Rana pretiosa) historically coexisted in portions of the Pacific Northwest now invaded by R. catesbeiana, but R. pretiosa has declined more severely than R. a. aurora. We investigated whether microhabitat and behavioral differences that facilitate sympatric coexistence of the natives predict which species is more susceptible to predation by introduced R. catesbeiana. Our laboratory experiments demonstrate that R. catesbeiana adults prefer aquatic microhabitats, that R. pretiosa juveniles are more aquatic than R. a. aurora, and that adult R. catesbeiana consume more R. pretiosa than R. a. aurora juveniles. Mean and maximum jump distances of R. pretiosa were shorter than equally sized R. a. aurora, and the difference between these two species increased with larger frog sizes. Our examination of field survey data indicates that R. pretiosa coexist with R. catesbeiana less frequently than R. a. aurora. We conclude that R. catesbeiana is a greater threat to survival of R. pretiosa than to R. a. aurora and suggest that microhabitat use and escape abilities of native ranid frogs may be linked to this asymmetrical effect. Analysis of behavioral and microhabitat differences among related native species may be a useful tool in predicting the effects of introduced predators on amphibians and can assist in developing conservation priorities for these species.