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Sample records for rana pipiens populations

  1. Prevalence of the pathogenic chytrid fungus, Batrachochytrium dendrobatidis, in an endangered population of northern leopard frogs, Rana pipiens

    PubMed Central

    2010-01-01

    Background Emerging infectious diseases threaten naïve host populations with extinction. Chytridiomycosis, an emerging infectious disease of amphibians, is caused by the pathogenic fungus Batrachochytrium dendrobatidis (Bd) and has been linked to global declines in amphibians. Results We monitored the prevalence of Bd for four years in the Northern leopard frog, Rana pipiens, which is critically imperiled in British Columbia (BC), Canada. The prevalence of Bd initially increased and then remained constant over the last three years of the study. Young of the year emerging from breeding ponds in summer were rarely infected with Bd. Some individuals cleared their Bd infections and the return rate between infected and uninfected individuals was not significantly different. Conclusions The BC population of R. pipiens appears to have evolved a level of resistance that allows it to co-exist with Bd. However, this small population of R. pipiens remains vulnerable to extinction. PMID:20202208

  2. Clinal patterns in genetic variation for northern leopard frog (Rana pipiens): Conservation status and population histories

    USGS Publications Warehouse

    Stockwell, Craig A.; Fisher, Justin D.L.; McLean, Kyle I.

    2016-01-01

    The security of the northern leopard frog (Rana pipiens) varies spatially with populations east and west of North Dakota considered as secure and at risk, respectively. We used genetic markers to characterize the conservation status of northern leopard frog populations across North Dakota. We used multiple regression analyses and model selection to evaluate correlations of expected heterozygosity (HE) with the direct and additive effects of: i) geographic location,ii) wetland density and iii) average annual precipitation. There was lower genetic diversity in the western portion of the state due to lower levels of diversity for populations southwest of the Missouri River. This may reflect a refugial/colonization signature for the only non-glaciated area of North Dakota. Genetic diversity was also positively associated with wetland densities which is consistent with the reliance of this species on a mosaic of wetlands. Our findings suggest that populations in the southwestern part of North Dakota are of higher conservation concern, a finding consistent with the higher risk noted for northern leopard frog populations in most states west of North Dakota. Our findings also pose the hypothesis that climate change induced changes in wetland densities will reduce genetic diversity of northern leopard frog populations.

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

    PubMed

    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

    2009-12-01

    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.

  4. 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.

  5. 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.

  6. Skin peptides protect juvenile leopard frogs (Rana pipiens) against chytridiomycosis.

    PubMed

    Pask, James D; Cary, Tawnya L; Rollins-Smith, Louise A

    2013-08-01

    One issue of great concern for the scientific community is the continuing loss of diverse amphibian species on a global scale. Amphibian populations around the world are experiencing serious losses due to the chytrid fungus, Batrachochytrium dendrobatidis. This pathogen colonizes the skin, leading to the disruption of ionic balance and eventual cardiac arrest. In many species, antimicrobial peptides secreted into the mucus are thought to contribute to protection against colonization by skin pathogens. Although it is generally thought that antimicrobial peptides are an important component of innate immune defenses against B. dendrobatidis, much of the current evidence relies on correlations between effective antimicrobial peptide defenses and species survival. There have been few studies to directly demonstrate that antimicrobial peptides play a role. Using the northern leopard frog, Rana pipiens, we show here that injection of noradrenaline (norepinephrine) brings about a long-term depletion of skin peptides (initial concentrations do not recover until after day 56). When peptide stores recovered, the renewed peptides were similar in composition to the initial peptides as determined by MALDI-TOF mass spectrometry and in activity against B. dendrobatidis as determined by growth inhibition assays. Newly metamorphosed froglets depleted of their peptide stores and exposed to B. dendrobatidis died more rapidly than B. dendrobatidis-exposed froglets with their peptides intact. Thus, antimicrobial peptides in the skin mucus appear to provide some resistance to B. dendrobatidis infections, and it is important for biologists to recognize that this defense is especially important for newly metamorphosed frogs in which the adaptive immune system is still immature.

  7. Acid exposure is an immune disruptor in adult Rana pipiens.

    PubMed

    Vatnick, Itzick; Andrews, Jaime; Colombo, Matthew; Madhoun, Hareth; Rameswaran, Muthuramanan; Brodkin, Marc A

    2006-01-01

    Acidic environments are physiological stressors for amphibians. The objective of the present study was to document the effect of an acidic environment on innate immune system function under controlled experimental conditions in Rana pipiens. We developed an in vivo assay, by injecting a suspension of 1-microm fluorescent beads in fluid thioglycollate, to induce peritonitis. The number of peritoneal exudate leukocytes and their phagocytic activity did not increase with thioglycollate injection when frogs were exposed to pH 5.5 compared to when frogs were exposed to pH 7.0. An environment of pH 5.5 disrupted the inflammatory response of frogs compared to an environment of pH 7.0; at pH 5.5, more nonphagocytic leukocytes and fewer highly phagocytic leukocytes were found compared to those in frogs exposed to pH 7.0. Frogs stimulated by thioglycollate injection and exposed to pH 5.5 had a 50% increase in cells that did not exhibit phagocytosis and a 4- to 10-fold reduction in the number of highly phagocytic cells. This is evidence that acid exposure functions as an immune disruptor in adult R. pipiens under laboratory conditions.

  8. 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.

  9. 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.

  10. Enterobacteriaceae and Aeromonas hydrophila in Minnesota frogs and tadpoles (Rana pipiens).

    PubMed Central

    Hird, D W; Diesch, S L; McKinnell, R G; Gorham, E; Martin, F B; Meadows, C A; Gasiorowski, M

    1983-01-01

    In 222 Rana pipiens frogs and 34 tadpoles captured in and near Minnesota, Aeromonas hydrophila and 29 species of Enterobacteriaceae, including yersinia enterocolitica and Salmonella arizonae, were isolated from intestines. The prevalence of members of the family Enterobacteriaceae was lowest in frogs captured in early spring and highest in frogs captured in late summer. PMID:6607034

  11. 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...

  12. 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...

  13. Ionic currents underlying the action potential of Rana pipiens oocytes.

    PubMed

    Schlichter, L C

    1989-07-01

    Ionic currents in immature, ovulated Rana pipiens oocytes (metaphase I) were studied using the voltage-clamp technique. At this stage of maturity the oocyte can produce action potentials in response to depolarizing current or as an "off response" to hyperpolarizing current. Reducing external Na+ to 1/10 normal (choline substituted) eliminated the action potentials and both the negative-slope region and zero-crossing of the I-V relation. Reducing external Cl- to 1/10 or 1/100 normal (methanesulfonate substituted) lengthened the action potential. The outward current was reduced and a net inward current was revealed. By changing external Na+, Cl-, and K+ concentrations and using blocking agents (SITS, TEA), three voltage- and time-dependent currents were identified, INa, IK and ICl. The Na+ current activated at about 0 mV and reversed at very positive values which decreased during maturation. Inward Na+ current produced the upstroke of the action potential. During each voltage-clamp step the Na+ current activated slowly (seconds) and did not inactivate within many minutes. The Na+ current was not blocked by TTX at micromolar concentrations. The K+ current was present only in the youngest oocytes. Because IK was superimposed on a large leakage current, it appeared to reverse at the resting potential. When leakage currents were subtracted, the reversal potential for IK was more negative than -110 mV in Ringer's solution. IK was outwardly rectifying and strongly activated above -50 mV. The outward K+ current produced an after hyperpolarization at the end of each action potential. IK was blocked completely and reversibly by 20 mM external TEA. The Cl- current activated at about +10 mV and was outwardly rectifying. ICl was blocked completely and reversibly by 400 microM SITS added to the bathing medium. This current helped repolarize the membrane following an action potential in the youngest oocytes and was the only repolarizing current in more mature oocytes that had lost

  14. 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.

  15. 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.

  16. Role for corticoids in mediating the response of Rana pipiens tadpoles to intraspecific competition.

    PubMed

    Glennemeier, Karen Ann; Denver, Robert John

    2002-01-01

    Competition is known to decrease growth and development rate in tadpoles, but the physiological basis for this phenomenon is poorly understood. We hypothesized that competition results in increased production of stress hormones and that these hormones are responsible for the suppression of growth and development. To test this hypothesis, we measured whole-body corticosterone content in premetamorphic Leopard frog (Rana pipiens) tadpoles raised at two different population densities and three different food levels. Whole body corticosterone content was elevated in tadpoles subjected to either limited food (at low density) or high density. Within the low and intermediate food treatments, high density reduced tadpole growth and slowed development. Limited food slowed growth and development at all densities. Blocking corticoid synthesis by treating tadpoles with metyrapone (MTP) reversed the growth suppression caused by high density (tested in the intermediate food level treatment) but did not alter the effect of density on development rate. MTP treatment did not alter the depressive effect of limited resources on growth or development. Our results suggest that elevated corticoid biosynthesis mediates the negative effect of increased population density (i.e., increased intraspecific competition) on tadpole growth. Copyright 2002 Wiley-Liss, Inc.

  17. Effects of agricultural pesticides on the immune system of Rana pipiens and on its resistance to parasitic infection.

    PubMed

    Christin, Marie-Soleil; Gendron, Andrée D; Brousseau, Pauline; Ménard, Lucie; Marcogliese, David J; Cyr, Daniel; Ruby, Sylvia; Fournier, Michel

    2003-05-01

    In the past 30 years, many amphibian species have suffered population declines throughout the world. Mass mortality have been frequently reported, and in several instances, infectious diseases appear to be the cause of death. The role that contaminants could play in these die-offs through immunotoxic effects has been poorly investigated. In this study, juvenile leopard frogs (Rana pipiens) were exposed for 21 d to a mixture of six pesticides (atrazine, metribuzin, aldicarb, endosulfane, lindane, and dieldrin) and subsequently challenged with a parasitic nematode, Rhabdias ranae. Exposure to the mixture at environmentally realistic concentrations significantly reduced lymphocyte proliferation. Three weeks after the end of the exposure, lymphocyte proliferation had recovered and was stimulated in frogs challenged with parasites with the exception of those previously exposed to the highest concentration. No pesticide effects on phagocytosis and splenocyte numbers were detectable at the end of the exposure period, but these two parameters were diminished 21 d after the infection challenge in frogs previously exposed to the highest levels of pesticides. In these animals, the prevalence of lung infection by R. ranae also tended to be higher. These results suggest that agricultural pesticides can alter the immune response of frogs and affect their ability to deal with parasitic infection.

  18. 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

  19. 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.

  20. Humane Anesthesia and Pain Management in Amphibian Limb Surgery of Rana pipiens

    PubMed Central

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

    2013-01-01

    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

  1. Electrical Signs of New Membrane Production during Cleavage of Rana pipiens Eggs

    PubMed Central

    Woodward, Donald J.

    1968-01-01

    Rana pipiens eggs dividing normally in diluted Ringer's solution show an increase in transmembrane potential inside negative, a decrease in resistance, and no change in total surface membrane capacitance at the appearance of a division furrow. Furrows of eggs in solutions with the tonicity of full Ringer develop partially, then regress so that the surface is again spherical. The potential and resistance changes are greater and substantial increases in capacitance occur when furrowing is so inhibited. It is proposed that the electrical changes at division are due to the introduction of new plasma membrane, between the blastomeres, having selective permeability to K and a low resistance compared to the outer spherical membrane. A narrow gap between blastomeres limits current flow through new membrane during normal division. A direct exposure of new membrane to the bathing medium when furrowing is disrupted results in larger changes in potential and resistance and permits the capacitance of new membrane to be detected. PMID:5691712

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

    PubMed

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

    1988-09-01

    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.

  3. Cutaneous transport of Ca2+ in the frog Rana pipiens: significance and specificity.

    PubMed

    Stiffler, D F; Eskandari, S; Dejbakhsh, S

    1997-04-01

    Rana pipiens were divided into four groups: controls; hypocalcemic frogs, depleted of salts by acclimation to deionized water; hypercalcemic frogs, calcium-loaded by the introduction of 40 mumol calcium gluconate; and frogs exposed to the potential competing ions Mg2+, Sr2+, and Ba2+. All groups displayed calcium influx that was proportional to external [Ca2+]; however, the group acclimated to deionized water also displayed hypocalcemia (P < 0.025) and enhanced Ca2+ influx at higher (> 0.3 mM) external [Ca2+]. Ca2+ efflux was depressed in hypocalcemic frogs, and thus net Ca2+ flux shifted from net loss in control frogs to net uptake in hypocalcemic frogs. Hypocalcemia also resulted in increased skin Ca2+ deposits which may be related to a decreased Ca2+ (and other ions) permeability as a consequence of the acclimation to deionized water. Another group of frogs was Ca(2+)-loaded by injecting calcium gluconate: Sodium gluconate controls did not significantly alter Ca2+ fluxes. The frogs that received calcium gluconate treatments became hypercalcemic (P < 0.01) and did not display significant changes in calcium fluxes, nor did they show significant changes in skin calcium deposits. We conclude that hypocalcemia leads to regulatory responses that stimulate active Ca2+ transport in Rana pipiens skin and possibly inhibits cutaneous and renal efflux. We also conclude that hypercalcemia does not alter calcium fluxes across skin. The ions from Group IIA of the Periodic Table of Elements had little effect on Ca2+ fluxes at concentrations ranging from 0.5-4.0 mM; neither Sr2+ or Ba2+ affected Ca2+ influx. The only divalent ion tested that influenced Ca2+ was Mg2+, which significantly inhibited Ca2+ influx but only at 4.0 mM or eight times the external [Ca2+]. We conclude, therefore, that the Ca2+ transport mechanism is fairly specific for Ca2+ within Group IIA.

  4. Status of RNAs, localized in Xenopus laevis oocytes, in the frogs Rana pipiens and Eleutherodactylus coqui.

    PubMed

    Nath, Kimberly; Boorech, Jamie L; Beckham, Yvonne M; Burns, Mary M; Elinson, Richard P

    2005-01-15

    Early development in the frog model, Xenopus laevis, is governed by RNAs, localized to the vegetal cortex of the oocyte. These RNAs include Xdazl RNA, which is involved in primordial germ cell formation, and VegT RNA, which specifies the mesoderm and endoderm. In order to determine whether orthologues of these RNAs are localized and have similar functions in other frogs, we cloned RpDazl and RpVegT from Rana pipiens, a frog that is phylogenetically distant from X. laevis. RNAs from both genes are localized to the vegetal cortex of the R. pipiens oocyte, indicating that the vegetal localization is likely the basal state. The animal location of EcVegT RNA in Eleutherodactylus coqui that we found previously (Beckham et al., 2003) is then a derived state, probably due to the great increase in egg size required for direct development of this species. To answer the question of function, we injected RpVegT or EcVegT RNAs into X. laevis embryos, and assayed animal caps for gene expression. Both of these RNAs induced the expression of endodermal, mesodermal, and organizer genes, showing that the function of RpVegT and EcVegT as meso-endodermal determinants is conserved in frogs. The RNA localizations and the function of VegT orthologues in germ layer specification may be synapomorphies for anuran amphibians.

  5. Vitellogenic cycles in laboratory-maintained females of the leopard frog, Rana pipiens.

    PubMed

    Smalley, K N; Nace, G W

    1983-05-01

    As a part of studies on the reproduction of laboratory maintained frogs, wild-caught Rana pipiens were ovulated and maintained at 22-27 degrees C for up to 18 months. Vitellogenic oocytes were periodically staged and counted, and a "maturity index" was calculated to assess the progress of the vitellogenic cycle. The initial cycle was similar to that of wild frogs except that the first oocytes to reach stage 5 (mature eggs) usually began to degenerate before later starting oocytes became mature. In addition, a second cycle began before the first was completed. After more than 1 year at room temperature, abnormal cycles were common. Ovaries of such animals contained very few mature eggs. Many of their oocytes were in early stages of vitellogenesis or, if pigmented, had begun to degenerate. These deficiencies were partially corrected in females placed in 4 degrees C for 4-6 weeks. The average number of mature eggs increased 15-fold and ovary weights more than doubled. Oviduct weights almost doubled. Although the rates of cooling, photoperiod, and nutritional status could be important influences, the results imply that cold treatment alone increases estrogen secretion. We suggest that low estrogen secretion may account for the reproductive deficiencies seen in R. pipiens cultured at room temperature.

  6. Morphological changes in the skin of Rana pipiens in response to metabolic acidosis.

    PubMed

    Page, R D; Frazier, L W

    1987-04-01

    The skin of Rana pipiens excretes H+ and this excretion is increased by metabolic acidosis. The mitochondria-rich (MR) cells of the skin have been found to mediate this H+ transport. The purpose of this study was to determine if there is a change in the MR cells of the skin during metabolic acidosis and if the isolated split epithelia of frog skin maintains its capacity to excrete H+. Metabolic acidosis was induced by injecting 120 mM NH4Cl (0.025 ml/g body wt) into the dorsal lymph sac three times a day for 2 days. The frogs were sacrificed and collagenase-split skins from the abdomen of normal and metabolic acidotic frogs were mounted between 2-ml chambers. H+ fluxes into both the mucosal and serosal media were measured and reported in units of (nmol) (cm2)-1 (min)-1. An increase in H+ flux was seen on both the mucosal and serosal sides of the acidotic split skins. The isolated epithelia were fixed, postosmicated, and dehydrated in the chamber. They were then embedded in Spurr's resin and 1-micron sections were cut and stained with Paragon multiple stain. Coded slides were used to count various cell types. Sections were randomly selected and approximately 40,000 cells were counted. Four basic cell types were noted and confirmed by TEM photomicrographs; basal (B) cells, granular (G) cells, keratinized cells, and MR cells. The ratio of G + B cells:MR cells in the normal skins was 1.0:0.021. The ratio in acidotic skins was 1.0:0.34. The average percentage of cell population of MR cells in the normal skins was 2.08 + 0.18 and in acidotic skins 3.20 + 0.36 (P less than 0.005). We conclude that the split skin maintains the capacity to acidify the mucosal fluid. Additionally, during metabolic acidosis there is an increased number of MR cells in the skin and this increase may be an adaptive mechanism of the skin to excrete excess H+ during acidosis.

  7. 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.

  8. Cryptic invasion of Northern Leopard Frogs (Rana pipiens) across phylogeographic boundaries and a dilemma for conservation of a declining amphibian

    USGS Publications Warehouse

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

    2017-01-01

    Anthropogenic introduction of species is a major contributor to loss of biodiversity. Translocations within the range of a species are less frequently recognized, but have the potential for negative effects as well. Genetic mixing may lead to loss of local adaptations or further decline through outbreeding depression. These cryptic invasions may be quite difficult to recognize, but genetic tools can be used to recognize and monitor such intraspecific introductions. Conversely, translocations within species can be an important conservation tool to reduce inbreeding depression and replace lost genetic diversity. Thus, cryptic invasions can be either an aid or a hindrance to conservation efforts. We tested for the presence of non-native genotypes and assessed the extent and nature of introgression in populations of Northern Leopard Frog (Rana pipiens) in the southwestern US, where populations have declined to a few remnant populations. The most abundant and diverse complex of populations in the region contained a mitochondrial haplotype that was not native to the western US, probably resulting from the introduction of released pets, laboratory animals, or release during fish stocking. These non-native haplotypes were well integrated into a large complex of ponds and lakes, contributing to high genetic diversity in this area. Logistically, the geographic extent of non-native genetic influence within this population precludes eliminating or controlling the non-native component of this population. We recommend assessing the progress and fate of the introgression over time—along with population fitness parameters—to determine whether this introduction is beneficial or detrimental to population persistence. Meanwhile, translocations from nearby locations with similar environmental conditions have the best prospects for avoiding problems with outbreeding depression in other declining populations and will also most effectively preserve regional genetic diversity.

  9. Circulation of marker substances in the cerebrospinal fluid of an amphibian, Rana pipiens.

    PubMed

    Jones, H C

    1980-01-01

    Solutions of fluorescein-labelled dextran or Evans blue-albumin were infused into the lateral cerebral ventricle of Rana pipiens. The subsequent distribution in the cerebrospinal fluid (CSF) was investigated between 2 and 24 h after infusion by freezing and examination of the cut blocks of the head and vertebral column of the stage of a freezing microtome. These marker substances move out of the ventricles into the subarachnoid space at the caudal end of the fourth ventricle and spread rapidly along the subarachnoid space of the spinal cord. The spreading of marker substances is slower into the brain subarachnoid space, When the marker is infused into the subarachnoid space of the forebrain, it becomes distributed throughout the subarachnoid space of the brain and spinal cord but not in the ventricles. Partial clearance of markers from the ventricles takes place within 5 h and total clearance within 8 h. Clearance from the brain and cord subarachnoid space is somewhat slower and can only be detected in experiments lasting 10 h or more. Absorption of the markers from the CSF occurs via the intervertebral foramina of the spinal cord. Fluorescence microscopy of sections of the cord show that the fluorescence leaves the subarachnoid space at the point where the spinal nerves traverse the arachnoid membrane.

  10. Haematoloechus sp. infection in wild-caught northern leopard frogs (Rana pipiens).

    PubMed

    Hsu, Charlie; Carter, D Bart; Williams, Donna; Besch-Williford, Cynthia

    2004-11-01

    Three male, wild-caught northern leopard frogs (Rana pipiens) died over a 1-week period with no previous history of clinical illness or disease. Noteworthy necropsy findings in one of the three frogs included depleted fat bodies in the coelomic cavity, indicating a poor nutritional condition, and a heavy parasite burden in the lungs. The location of infection and morphologic characteristics of the parasite were consistent with infection by the common lung fluke, Haematoloechus sp. In contrast to the heavy fluke load, only minor microscopic changes were observed in the lungs. Lesions included mild hypertrophy of the bronchiolar epithelium, with few submucosal inflammatory cells consisting predominantly of lymphocytes. Subsequent review of the literature revealed little about the pathologic effects of these parasites except that small numbers are thought to cause the host little harm. Our findings suggest that even with a large number of parasites, there is minimal pathologic impact in the lungs. We conclude that heavy lung-fluke infection should not be diagnosed as the sole or major etiology of death or illness in leopard frogs.

  11. Effects of exposure to cold on metabolic characteristics in gastrocnemius muscle of frog (Rana pipiens).

    PubMed Central

    Ohira, M; Ohira, Y

    1988-01-01

    1. Responses of enzymic characteristics of gastrocnemius muscle were studied when frogs (Rana pipiens) were exposed to cold environment (4 degrees C). 2. The content of adenosine triphosphate (ATP) decreased significantly after cold exposure. This decrease was greater in starved than in fed frogs. 3. Although the glycogen content did not change, lactate levels were lower in cold-exposed than room-temperature (control) frogs. No change was observed in glycogen and lactate between fed and unfed frogs kept at 4 degrees C for 2 months. Lactate dehydrogenase activity tended to increase during chronic cold exposure, but not significantly. 4. The activities of citrate synthase, cytochrome oxidase, and beta-hydroxyacyl CoA dehydrogenase were higher in gastrocnemius of chronically cold-exposed frogs than in room-temperature controls. This increase was statistically significant only in the muscles of starved frogs; these muscles had the greatest decrease in ATP. 5. It was suggested that chronic cold exposure decreases skeletal muscle ATP content but may not affect glycolysis. The data also suggested that the decrease in ATP content stimulates mitochondrial biogenesis which increases enzyme activities. PMID:3261790

  12. Apoptosis in the digestive tract of herbivorous Rana pipiens larvae and carnivorous Ceratophrys ornata larvae: an immunohistochemical study.

    PubMed

    Kaltenbach, Jane C; Fry, Anne E; Colpitts, Katherine M; Faszewski, Ellen E

    2012-01-01

    The lifespan of herbivorous Rana pipiens larvae is ∼3 months, while that of carnivorous Ceratophrys ornata larvae is only about 2 weeks. During metamorphic climax, the larval gut shortens dramatically, especially in R. pipiens, and its luminal epithelium is replaced by adult-type epithelium. To determine when programmed cell death occurs during the metamorphic restructuring of the gut, we prepared cross-sections of the stomach, small intestine, and large intestine from representative larval stages and from juvenile frogs of both species. The sections were incubated with monoclonal antibody against active caspase-3, one of the key enzymes in the apoptotic cascade. We observed apoptosis in some luminal epithelial cells in each of the three regions of the larval gastrointestinal tract of both species. However, apoptotic cells appeared earlier in larval stages of R. pipiens than C. ornata and few were seen in juvenile frogs of either species. The results demonstrate the occurrence of apoptosis in the metamorphic remodeling of the gut of both R. pipiens larvae and C. ornata larvae. Copyright © 2011 Wiley Periodicals, Inc.

  13. Light-dependent toxicity of α-terthienyl and anthracene toward late embryonic stages ofRana pipiens.

    PubMed

    Kagan, J; Kagan, P A; Buhse, H E

    1984-07-01

    Alpha-terthienyl is toxic to late embryonic stages ofRana pipiens in the presence of sunlight. Neither α-terthienyl alone in the dark nor a previously photolyzed solution of α-terthienyl has comparable activity. The LC50 was 0.11 ppm with 30 min of exposure and 0.018 ppm with 2 hr of exposure to sunlight. Anthracene, a representative example of polycyclic aromatic hydrocarbons widely distributed in the environment, also showed similar phototoxicity, with an LC50 of 0.065 ppm after 30 min of exposure and 0.025 ppm after 5 hr.

  14. Potential endocrine disruption of sexual development in free ranging male northern leopard frogs (Rana pipiens) and green frogs (Rana clamitans) from areas of intensive row crop agriculture.

    PubMed

    McDaniel, Tana V; Martin, Pamela A; Struger, John; Sherry, Jim; Marvin, Chris H; McMaster, Mark E; Clarence, Stacey; Tetreault, Gerald

    2008-07-30

    Intensive row crop agriculture (IRCA) for corn and soybean production is predominant in eastern and central North America. IRCA relies heavily on pesticide and nutrient inputs to maximize production under conventional systems. In 2003-2005, we assessed the occurrence of a suite of potential endocrine effects in amphibians inhabiting farm ponds and agricultural drains in IRCA areas of southwestern Ontario. Effects were compared to amphibians from two agricultural reference sites as well as four non-agricultural reference sites. Pesticide and nutrient concentrations were also determined in water samples from those sites. Atrazine and metolachlor were detected in most samples, exceeding 1 microg L(-1) at some sites. Blood samples were taken from northern leopard frogs (Rana pipiens) and green frogs (Rana clamitans) for analysis of circulating sex steroids and vitellogenin-like protein (Vtg-lp), a biomarker of exposure to environmental estrogens. Gonads were histologically examined for evidence of abnormalities. Some evidence of exposure to endocrine disrupting compounds was apparent from the data. The occurrence of testicular ovarian follicles (TOFS) in male R. pipiens was significantly higher (42%; p<0.05) at agricultural sites, particularly those in Chatham county compared to frogs from reference sites (7%). There was no difference in circulating sex steroid levels between frogs from agricultural and reference sites and sex steroid levels did not correlate with pesticide concentrations in the environment. No differences were detected in the gonadosomatic indices or stage of spermatogenesis between frogs from agricultural and non-agricultural regions (p>0.05). Plasma Vtg-lp was detected in only one male R. pipiens from an agricultural site. Neither gonad size, gonad maturity nor sex steroid levels differed between normal males and those with testicular oocytes. Although the proportion of testicular oocytes did not correlate directly with atrazine concentrations, it

  15. Sensory transduction in dorsal cutaneous mechanoreceptors of the frog, Rana pipiens.

    PubMed

    Watts, R E; French, A S

    1985-11-01

    Sensory transduction was studied in dorsal skin mechanoreceptors of the frog, Rana pipiens. The skin was clamped and stretched before being stimulated with the tip of a glass rod mounted on a servo-controlled loudspeaker. Afferent activity was recorded extracellularly from a dorsal cutaneous nerve. Three groups of sensory units could be identified by the size of their recorded action potentials and their response to mechanical stimuli. Action potential amplitudes for the three groups were: less than 50 microV (group I), 50-250 microV (group II) and greater than 300 microV (group III). Group II were selected for further study because of their amplitude and their resistance to fatigue. Three types of mechanical stimuli were used to examine the dynamic properties of group II receptors, steps, sinusoids, and band-limited random displacement. In each case the responses could be well fitted by a power-law model with a fractional exponent of time or frequency. Random stimulation of a large number of group II receptors showed considerable variability in their sensitivity and in their dynamic behavior, as measured by the fractional exponent of frequency. However, the distributions of these two parameters were both unimodal and strongly clustered around the modes, suggesting that the recordings were from a single class of receptors. Varying the temperature of the receptors had little effect on their sensitivity or dynamic properties. This is in contrast to findings on other mechanoreceptors. Doubling the potassium concentration in the bathing solution affected the dynamic properties of the receptors within 5 min but several distinct patterns of change in dynamic behavior were seen.(ABSTRACT TRUNCATED AT 250 WORDS)

  16. Evaluation of metomidate hydrochloride as an anesthetic in leopard frogs (Rana pipiens).

    PubMed

    Doss, Grayson A; Nevarez, Javier G; Fowlkes, Natalie; da Cunha, Anderson F

    2014-03-01

    Metomidate hydrochloride is an imidazole-based, nonbarbiturate hypnotic drug primarily used as an immersion sedation and anesthetic agent in freshwater and marine finfish. To the authors' knowledge, there is no documentation in the literature of its use in amphibians. In this study, 7 male and 4 female leopard frogs (Rana pipiens) were induced with metomidate hydrochloride via immersion bath at a concentration of 30 mg/L for 60 min. The pH of the induction solution ranged from 7.63 to 7.75. Each frog was then removed from the induction solution, rinsed, and recovered in 26.6 degrees C amphibian Ringer's solution. After 210 min in the Ringer's solution, the frogs were transferred to moist paper towels for recovery. Heart rate, gular and abdominal respiration rates, righting reflex, superficial and deep pain withdrawal reflexes, corneal and palpebral reflexes, and escape response were monitored and recorded at defined intervals during both induction and recovery. The average time to loss of righting reflex and escape response was 17.36 min and 17.82 min, respectively. Metomidate produced clinical sedation in all frogs (n = 11). Surgical anesthesia was achieved in only 27% (3/11), with an anesthetic duration that ranged from 9 to 20 min. Recovery times were extremely prolonged and varied, with a range from 313 min to longer than 600 min. The findings of this study indicate that metomidate hydrochloride is unsuitable as a sole anesthetic agent in leopard frogs, and further research is needed to evaluate its suitability in other amphibians.

  17. 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

  18. 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

  19. 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...

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

    USDA-ARS?s Scientific Manuscript database

    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...

  1. 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...

  2. Atrazine-induced hermaphroditism at 0.1 ppb in American leopard frogs (Rana pipiens): laboratory and field evidence.

    PubMed

    Hayes, Tyrone; Haston, Kelly; Tsui, Mable; Hoang, Anhthu; Haeffele, Cathryn; Vonk, Aaron

    2003-04-01

    Atrazine is the most commonly used herbicide in the United States and probably the world. Atrazine contamination is widespread and can be present in excess of 1.0 ppb even in precipitation and in areas where it is not used. In the current study, we showed that atrazine exposure (> or = to 0.1 ppb) resulted in retarded gonadal development (gonadal dysgenesis) and testicular oogenesis (hermaphroditism) in leopard frogs (Rana pipiens). Slower developing males even experienced oocyte growth (vitellogenesis). Furthermore, we observed gonadal dysgenesis and hermaphroditism in animals collected from atrazine-contaminated sites across the United States. These coordinated laboratory and field studies revealed the potential biological impact of atrazine contamination in the environment. Combined with reported similar effects in Xenopus laevis, the current data raise concern about the effects of atrazine on amphibians in general and the potential role of atrazine and other endocrine-disrupting pesticides in amphibian declines.

  3. Atrazine-induced hermaphroditism at 0.1 ppb in American leopard frogs (Rana pipiens): laboratory and field evidence.

    PubMed Central

    Hayes, Tyrone; Haston, Kelly; Tsui, Mable; Hoang, Anhthu; Haeffele, Cathryn; Vonk, Aaron

    2003-01-01

    Atrazine is the most commonly used herbicide in the United States and probably the world. Atrazine contamination is widespread and can be present in excess of 1.0 ppb even in precipitation and in areas where it is not used. In the current study, we showed that atrazine exposure (> or = to 0.1 ppb) resulted in retarded gonadal development (gonadal dysgenesis) and testicular oogenesis (hermaphroditism) in leopard frogs (Rana pipiens). Slower developing males even experienced oocyte growth (vitellogenesis). Furthermore, we observed gonadal dysgenesis and hermaphroditism in animals collected from atrazine-contaminated sites across the United States. These coordinated laboratory and field studies revealed the potential biological impact of atrazine contamination in the environment. Combined with reported similar effects in Xenopus laevis, the current data raise concern about the effects of atrazine on amphibians in general and the potential role of atrazine and other endocrine-disrupting pesticides in amphibian declines. PMID:12676617

  4. Absorption of the cerebrospinal fluid and intracranial compliance in an amphibian, Rana pipiens.

    PubMed

    Jones, H C; Taylor, C M

    1984-08-01

    Experiments have been carried out to investigate the absorption of the cerebrospinal fluid (c.s.f.) and the intracranial compliance in an amphibian, Rana pipiens, using infusions into the c.s.f. system through glass micropipettes. Resistance to absorption of the c.s.f. was estimated by the constant rate infusion technique. Mean absorption resistance for infusions of artificial c.s.f. into the lateral ventricles and into the cerebral subarachnoid space were 15.48 and 16.52 mmH2O min microliter-1 respectively. This difference was not significant and it is concluded that the pores in the posterior tela situated in the roof of the fourth ventricle do not offer any resistance to the flow of c.s.f. out of the ventricles. The resistance to drainage of the c.s.f. in this amphibian is higher than that found for mammals. Mean resting c.s.f. pressure, estimated from the intercept of the regression line with the pressure axis at zero infusion rate was 18.0 mmH2O. Absorption resistance was measured from the cerebral subarachnoid space before and after injection of 4 microliter Indian ink solution. There was a 3-fold increase in resistance following ink injection. Two-way analysis of variance showed the difference to be significant (P less than 0.01) suggesting that the outflow sites can become partially blocked by particulate matter. During a continuous 3 h infusion of artificial c.s.f. containing [14C]dextran or [125I]-labelled human serum albumin (RISA) into the lateral ventricles, the mean percentage uptakes into the systemic circulation after the first 0.5 h of a 3 h period were 74.1 and 61.9% respectively. The difference is not significant. The rapid and high uptake into blood suggests there is a direct communication between c.s.f. and blood in amphibians. During continuous infusion of RISA into the lateral ventricles, simultaneous blood samples were taken from the femoral artery and the internal dorsal vertebral vein. Radioactivity was found to be 13.2% higher in venous

  5. 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. © 2014 SETAC.

  6. 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.

  7. The effects of four arthropod diets on the body and organ weights of the leopard frog, Rana pipiens, during vitellogenesis.

    PubMed

    Lehman, G C

    1978-12-01

    Wild-caught adult Rana pipiens females were captured in midsummer and fed diets of crickets, flies sowbugs or wax moth larvae during a three-month period of active vitellogenesis. The cricket diet supported the most extensive body weight gain during this time and promoted a prolonged period of weight increase in an additional long-term study. Synchronous growth of the oocytes occurred in all four groups, but the ovaries and oviducts of cricket-fed animals were significantly larger than those of frogs on the other three diets. The significantly higher liver weights of frogs fed wax moth larvae may have reflected an augmentation of hepatic energy stores. Fat body weights were also highest in this group of animals. Frogs fed crickets and wax moth larvae possessed larger fat bodies than did the midsummer control animals killed immediately after their arrival in the laboratory. In contrast, frogs fed flies and sowbugs had smaller fat bodies than did the initial controls, suggesting that animals on these diets had utilized fat body lipid during vitellogenesis. Gastrocnemius and final body weights were lowest in frogs fed wax moth larvae. These findings may have reflected the nutritional content of the diet or the reduction in appetite frequently noted in these animals during observations of feeding behavior.

  8. The physiological effects of hydrostatic pressure are not equivalent to those of helium pressure on Rana pipiens.

    PubMed Central

    Dodson, B A; Furmaniuk, Z W; Miller, K W

    1985-01-01

    The effects of helium pressure and hydrostatic pressure on Rana pipiens were compared. Both agents caused paralysis at pressures greater than 135 atmospheres (1 atm = 101.325 kPa), but the median pressure for hydrostatic-pressure-induced paralysis was 35 atm less than that for helium pressure. When the ability of both pressurizing agents to reverse urethane-induced anaesthesia was compared, it was found that hydrostatic pressure raised the median dose for anaesthesia 2.2-fold more per atmosphere than did helium pressure. Animals that were lightly anaesthetized by urethane at 110 atm hydrostatic pressure became more deeply anaesthetized when helium was admitted isobarically into the pressure chamber. This difference in depth of anaesthesia between hydrostatic pressure and helium pressure is consistent with helium possessing an inherent anaesthetic effect. The abilities of other gases to pressure-reverse urethane anaesthesia were also determined. The degree of attenuation of the full pressure reversal effect observed with hydrostatic pressure was proportional to the lipid solubility of the gases, increasing in the order helium, neon, hydrogen, nitrogen and argon. Our data on the difference between hydrostatic and helium pressure are consistent with the critical volume hypothesis. PMID:3874954

  9. 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.

  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. 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. © 2015 SETAC.

  12. 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

  13. 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.

  14. Small frogs get their worms first: the role of nonodonate arthropods in the recruitment of Haematoloechus coloradensis and Haematoloechus complexus in newly metamorphosed northern leopard frogs, Rana pipiens, and woodhouse's toads, Bufo woodhousii.

    PubMed

    Bolek, Matthew G; Janovy, John

    2007-04-01

    Studies on the life cycles and epizootiology of North American frog lung flukes indicate that most species utilize odonates as second intermediate hosts; adult frogs become infected by ingesting odonate intermediate hosts. Newly metamorphosed frogs are rarely infected with these parasites, predominantly because they are gape-limited predators that cannot feed on large intermediate hosts such as dragonflies. We examined the role of the frog diet and potential intermediate hosts in the recruitment of the frog lung fluke, Haematoloechus coloradensis, to metamorphosed northern leopard frogs (Rana pipiens), Woodhouse's toads (Bufo woodhousii), and bullfrogs (Rana catesbeiana) from western Nebraska. Because of the uncertain validity of H. coloradensis as a distinct species from Haematoloechus complexus, morphological characters of both species were reevaluated and the life cycles of both species were completed in the laboratory. The morphological data on H. coloradensis and H. coimplexus indicate that they differ in their oral sucker to pharynx ratio, uterine loop distribution, and placement of vitelline follicles. However, in terms of their life cycles, both species are quite similar in their use of physid snails as first intermediate hosts, a wide range of nonodonate and odonate arthropods as second intermediate hosts, and leopard frogs and toads as definitive hosts. These results indicate that H. coloradensis and H. complexus are generalists at the second intermediate host level and might be able to infect newly metamorphosed leopard frogs and toads by using small nonodonate arthropods more commonly than other frog lung fluke species. Comparisons of population structure of adult flukes in newly metamorphosed leopard frogs indicate that the generalist nature of H. coloradensis metacercariae enables it to colonize young of the year leopard frogs more commonly than other Haematoloechus spp. that only use odonates as second intermediate hosts. In this respect, the

  15. Low levels of the herbicide atrazine alter sex ratios and reduce metamorphic success in Rana pipiens tadpoles raised in outdoor mesocosms.

    PubMed

    Langlois, Valérie S; Carew, Amanda C; Pauli, Bruce D; Wade, Michael G; Cooke, Gerard M; Trudeau, Vance L

    2010-04-01

    There are conflicting reports regarding the effects of atrazine (ATZ) on amphibian development. Therefore, further studies are needed to examine the potential mechanisms of action of ATZ in amphibians. Our aim in this study was to determine whether low concentrations of ATZ affect gonadal development and metamorphosis in the Northern leopard frog, Rana pipiens. Tadpoles were exposed in outdoor mesocosms to nominal concentrations of 0.1 and 1.8 microg/L of formulated ATZ from Gosner stage 27 (G27) to metamorphic climax (G42). Exposure to 17alpha-ethinylestradiol (EE2; 1.5 microg/L) provided a positive control for induction of testicular oocytes in males. Endocrine-related gene expression and gonadal histopathology were examined at G42 and in a subset of premetamorphic G34 tadpoles that failed to metamorphose. Gonadal gross morphology revealed that the 1.8-microg/L ATZ treatment produced 20% more females compared with the control. Histologic analysis revealed that 22% of EE2-treated males had testicular oocytes, whereas none were observed in any animals from the control or either ATZ groups. ATZ increased brain estrogen receptor alpha mRNA to 2.5 times that of the control at premetamorphosis and altered liver levels of 5beta-reductase activity at metamorphosis. In contrast, brain aromatase mRNA level and activity did not change. ATZ treatments significantly reduced metamorphic success (number of animals reaching metamorphosis) without affecting body weight, snout-vent length, or age at metamorphosis. Gene expression analysis indicated that ATZ decreased the expression of deiodinase type 3 in the tail at premetamorphosis. Our study indicates that exposure to low concentrations of ATZ in experimental mesocosms alters gonadal differentiation and metamorphosis in developing R. pipiens.

  16. Isolation and characterization of viruses from the kidneys of Rana pipiens with renal adenocarcinoma before and after passage in the red eft (Triturus viridescens).

    PubMed

    Clark, H F; Brennan, J C; Zeigel, R F; Karzon, D T

    1968-06-01

    Viruses were isolated from kidneys of normal and renal tumor-bearing Vermont Rana pipiens after subinoculation into red eft newts (Triturus viridescens). Organs of efts inoculated with viable cell suspensions from four of seven tumor-bearing kidneys yielded virus (LT-1, -2, -3, -4) when inoculated into TH-1 (Terrapene heart) cell culture. One tumor-bearing kidney also yielded virus (L-4) by direct inoculation into TH-1 cells. An additional isolate (L-5) was obtained from 1 of 52 normal Vermont frog kidneys inoculated directly into TH-1 cells. LT-1 was propagated with cytopathic effect (CPE) in each of 38 cell types tested, of fish, amphibian, reptilian, avian, and mammalian origin, at 23 or 30 C. LT-1 through LT-4, L-4 and L-5, and FV-1 through FV-3 each induced similar CPE in all cells tested. LT-2, however, induced CPE that progressed at a slower rate than that caused by the other isolates and produced smaller plaques (<0.8 mm) under starch gel overlay. Each of the viruses replicated to high titer in embryonated eggs incubated at 30 C. The viruses also grew in efts and adult newts, but not in bullfrog (Rana catesbeiana) tadpoles or adult leopard frogs. Tumor induction in adult leopard frogs inoculated with LT-1 was not demonstrated. Electron microscopic observations of LT-1 and LT-2 viruses revealed cytoplasmic particles, hexagonal in cross section, approximately 120 to 140 mmu in diameter, containing a dense nucleoid. LT-1 and LT-2 viruses were indistinguishable from FV-1 and Tipula iridescent virus. LT-1 was presumed to be a deoxyribonucleic acid virus on the basis of 5-bromodeoxyuridine inhibition. The isolates were ether-sensitive. On the basis of biological, physicochemical, and antigenic similarities, LT-1 through LT-4, L-4, L-5, FV-1 through FV-3, and isolates recently recovered from the bullfrog and the newt may represent strains of the same amphibian cytoplasmic virus.

  17. Gonadotropin stimulation of steroid synthesis and metabolism in the Rana pipiens ovarian follicle: sequential changes in endogenous steroids during ovulation, fertilization and cleavage stages.

    PubMed

    Morrill, Gene A; Schatz, Frederick; Kostellow, Adele; Bloch, Eric

    2006-05-01

    Steroid synthesis and metabolism have been followed in Rana pipiens ovarian follicles, denuded oocytes and eggs during ovulation, fertilization and cleavage stages (blastula formation). Under physiological conditions, gonadotropin stimulation of the fully grown follicle leads to progesterone synthesis from [(3)H]acetate as well as formation of much smaller amounts of 17alpha-hydroxyprogesterone, androstenedione, pregnanedione and pregnanediol. Progesterone levels increase during completion of the first meiotic division, but by ovulation progesterone disappears from the egg. Plasma membrane-bound progesterone is taken up into the oocyte cortical granules and is largely metabolized to 5alpha-pregnane-3alphaol,20-one and 5beta-pregnane-3alpha,17alpha,20beta-triol coincident with internalization of 60% of the oocyte surface (and >90% of bound progesterone) by the end of the hormone-dependent period. The principal steroid in the ovulated egg is 5beta-pregnane-3alpha,17alpha,20beta-triol. There is a rapid efflux of 5beta-pregnane-3alpha,17alpha,20beta-triol into the medium immediately following fertilization and residual steroid levels remain low in the developing blastula. Dissociated blastulae cells prepared from stage 9 1/2 embryos concentrate both pregnenolone and progesterone from the medium with minimal metabolism. The results indicate that the ovarian follicle has the ability to synthesize and metabolize progesterone but that this ability disappears in the ovulated egg. The progesterone metabolites formed during meiosis are largely released at fertilization.

  18. 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.

  19. 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.

  20. Genetic diversity of Culex pipiens mosquitoes in distinct populations from Europe: contribution of Cx. quinquefasciatus in Mediterranean populations.

    PubMed

    Shaikevich, Elena V; Vinogradova, Elena B; Bouattour, Ali; Gouveia de Almeida, António Paulo

    2016-01-27

    Mosquitoes of the Culex pipiens complex are cosmopolitan, and important vectors of neglected tropical diseases, such as arbovirosis and lymphatic filariasis. Among the complex taxa, Cx. pipiens (with two forms pipiens and molestus) and Cx. quinquefasciatus are the most ubiquitous mosquitoes in temperate and tropical regions respectively. Mosquitoes of this taxa lack of morphological differences between females, but have frank behavioral and physiological differences and have different trophic preferences that influence their vectorial status. Hybridization may change the vectorial capacity of these mosquitoes, increasing vector efficiency and medical importance of resulting hybrids. Culex pipiens s.l. from 35 distinct populations were investigated by the study of mtDNA, symbiotic bacterium Wolbachia pipientis, nuclear DNA and flanking region of microsatellite CQ11 polymorphism using PCR with diagnostic primers, RFLP analysis and sequencing. Six different mitochondrial haplotypes were revealed by sequencing of the cytochrome oxidase subunit I (COI) gene and three different Wolbachia (wPip) groups were identified. A strong association was observed between COI haplotypes/groups, wPip groups and taxa; haplogroup A and infection with wPipII appear to be typical for Cx. pipiens form pipiens, haplotype D and infection with wPipIV for form molestus, while haplogroup E, characteristic of Cx. quinquefasciatus, were correlated with wPipI and found in Cx. pipiens sl. from coastal regions of Southern Europe and Mediterranean region. Analysis of microsatellite locus and nuclear DNA revealed hybrids between Cx. pipiens form pipiens and form molestus, as well as between Cx. pipiens and Cx. quinquefasciatus, in Mediterranean populations, as opposed to Northern Europe. Phylogenetic analysis of COI sequences yielded a tree topology that supported the RFLP analysis with significant bootstrap values for haplotype D and haplogroup E. Molecular identification provides the first evidence

  1. Population size, survival, growth, and movements of Rana sierrae

    USGS Publications Warehouse

    Fellers, Gary M.; Kleeman, Patrick M.; Miller, David A. W.; Halstead, Brian J.; Link, William

    2013-01-01

    Based on 2431 captures of 757 individual frogs over a 9-yr period, we found that the population of R. sierrae in one meadow–stream complex in Yosemite National Park ranged from an estimated 45 to 115 adult frogs. Rana sierrae at our relatively low elevation site (2200 m) grew at a fast rate (K = 0.73–0.78), had high overwintering survival rates (44.6–95%), lived a long time (up to 16 yr), and tended to be fairly sedentary during the summer (100% minimum convex polygon annual home ranges of 139 m2) but had low year-to-year site fidelity. Even though the amphibian chytrid fungus (Batrachochytrium dendrobatidis, Bd) has been present in the population for at least 13 yr, there was no clear downward trend as might be expected from reports of R. sierrae population declines associated with Bd or from reports of widespread population decline of R. sierrae throughout its range.

  2. Assessment of the risk of solar ultraviolet radiation to amphibians. I. Dose-dependent induction of hindlimb malformations in the northern leopard frog (Rana pipiens).

    PubMed

    Ankley, Gerald T; Diamond, Stephen A; Tietge, Joseph E; Holcombe, Gary W; Jensen, Kathleen M; Defoe, David L; Peterson, Ryan

    2002-07-01

    A number of environmental stressors have been hypothesized as responsible for recent 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 ultraviolet (UV) radiation in causing limb malformations in a species in which this phenomenon seemingly is particularly prevalent, the northern leopard frog (Rana pipiens). Frogs were exposed from early embryonic stages through complete metamorphosis to varying natural sunlight regimes, including unaltered (100%) sunlight, sunlight subjected to neutral density filtration to achieve relative intensities of 85%, 75%, 65%, 50%, and 25% of unaltered sunlight, and sunlight filtered with glass or acrylamide to attenuate, respectively, the UVB (290-320 nm) and UVB plus UVA (290-380 nm) portions of the spectrum. The experiments were conducted in a controlled setting, with continual monitoring of UVB, UVA, and visible light to support a robust exposure assessment. Full sunlight caused approximately 50% mortality of the frogs during early larval development; no significant treatment-related mortality occurred under any of the other exposure regimes, including 100% sunlight with glass or acrylamide filtration. There was a dose-dependent (p < 0.0001) induction of hindlimb malformations in the frogs, with the percentage of affected animals ranging from about 97% under unaltered sunlight to 0% in the 25% neutral density treatment. Malformations were comprised mostly of missing or truncated digits, and generally were bilateral as well as symmetrical. Filtration of sunlight with either glass or acrylamide both significantly reduced the incidence of malformed limbs. The estimated sunlight dose resulting in a 50% limb malformation rate (ED50) was 63.5%. The limb ED50 values based on measured sunlight intensities corresponded to average daily doses of 4.5 and 100 Wh x m(-2) for UVB and UVA, respectively

  3. 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

  4. Estrogenic exposure affects metamorphosis and alters sex ratios in the northern leopard frog (Rana pipiens): identifying critically vulnerable periods of development.

    PubMed

    Hogan, Natacha S; Duarte, Paula; Wade, Michael G; Lean, David R S; Trudeau, Vance L

    2008-05-01

    During the transformation from larval tadpole to juvenile frog, there are critical periods of metamorphic development and sex differentiation that may be particularly sensitive to endocrine disruption. The aim of the present study was to identify sensitive developmental periods for estrogenic endocrine disruption in the northern leopard frog (Rana pipiens) using short, targeted exposures to the synthetic estrogen, ethinylestradiol (EE2). Post-hatch tadpoles (Gosner stage 27) were exposed over five distinct periods of metamorphosis: early (stage 27-30), mid (stage 30-36), early and mid (stage 27-36), late (stage 36-42), and the entire metamorphic period (chronic; stage 27-42). For each period, animals were sampled immediately following the EE2 exposure and at metamorphic climax (stage 42). The effects of EE2 on metamorphic development and sex differentiation were assessed through measures of length, weight, developmental stage, days to metamorphosis, sex ratios and incidence of gonadal intersex. Our results show that tadpoles exposed to EE2 during mid-metamorphosis were developmentally delayed immediately following exposure and took 2 weeks longer to reach metamorphic climax. In the unexposed groups, there was low proportion (0.15) of intersex tadpoles at stage 30 and gonads appeared to be morphologically distinct (male and female) in all individuals by stage 36. Tadpoles exposed early in development displayed a strong female-biased sex ratio compared to the controls. Moreover, these effects were also seen at metamorphic climax, approximately 2-3 months after the exposure period, demonstrating that transient early life-stage exposure to estrogen can induce effects on the reproductive organs that persist into the beginning of adult life-stages.

  5. 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

  6. Modeling dynamics of culex pipiens complex populations and assessing abatement strategies for West Nile Virus.

    PubMed

    Pawelek, Kasia A; Niehaus, Patrick; Salmeron, Cristian; 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.

  7. 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...

  8. 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...

  9. 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

  10. 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

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

    2013-12-01

    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.

  11. Multiple Wolbachia determinants control the evolution of cytoplasmic incompatibilities in Culex pipiens mosquito populations.

    PubMed

    Atyame, Celestine M; Duron, Olivier; Tortosa, Pablo; Pasteur, Nicole; Fort, Philippe; Weill, Mylene

    2011-01-01

    Wolbachia are maternally inherited endosymbionts that can invade arthropod populations through manipulation of their reproduction. In mosquitoes, Wolbachia induce embryonic death, known as cytoplasmic incompatibility (CI), whenever infected males mate with females either uninfected or infected with an incompatible strain. Although genetic determinants of CI are unknown, a functional model involving the so-called mod and resc factors has been proposed. Natural populations of Culex pipiens mosquito display a complex CI relationship pattern associated with the highest Wolbachia (wPip) genetic polymorphism reported so far. We show here that C. pipiens populations from La Réunion, a geographically isolated island in the southwest of the Indian Ocean, are infected with genetically closely related wPip strains. Crossing experiments reveal that these Wolbachia are all mutually compatible. However, crosses with genetically more distant wPip strains indicate that Wolbachia strains from La Réunion belong to at least five distinct incompatibility groups (or crossing types). These incompatibility properties which are strictly independent from the nuclear background, formally establish that in C. pipiens, CI is controlled by several Wolbachia mod/resc factors.

  12. 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

  13. 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

  14. Impacts of agriculture on the parasite communities of northern leopard frogs (Rana pipiens) in southern Quebec, Canada.

    PubMed

    King, K C; McLaughlin, J D; Gendron, A D; Pauli, B D; Giroux, I; Rondeau, B; Boily, M; Juneau, P; Marcogliese, D J

    2007-12-01

    Given that numerous amphibians are suffering population declines, it is becoming increasingly important to examine the relationship between disease and environmental disturbance. Indeed, while many studies relate anthropogenic activity to changes in the parasitism of snails and fishes, little is known of the impact on the parasites of amphibians, particularly from agriculture. For 2 years, the parasite communities of metamorphic northern leopard frogs from 7 agricultural wetlands were compared with those from 2 reference wetlands to study differences in parasite community diversity and abundance of various species under pristine conditions and 3 categories of disturbance: only agricultural landscape, only pesticides, and agricultural landscape with pesticides. Agricultural (and urban) area was negatively related to species richness, and associated with the near absence of adult parasites and species that infect birds or mammals. We suggest that agriculture and urbanization may hinder parasite transmission to frogs by limiting access of other vertebrate hosts of their parasites to wetlands. The only parasite found at all localities was an unidentified echinostome infecting the kidneys. This parasite dominated communities in localities surrounded by the most agricultural land, suggesting generalist parasites may persist in disrupted habitats. Community composition was associated with dissolved organic carbon and conductivity, but few links were found with pesticides. Pollution effects may be masked by a strong impact of land use on parasite transmission.

  15. 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...

  16. 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...

  17. Characterization of Culex pipiens Complex (Diptera: Culicidae) Populations in Colorado, USA Using Microsatellites

    PubMed Central

    Kothera, Linda; Godsey, Marvin S.; Doyle, Michael S.; Savage, Harry M.

    2012-01-01

    Mosquitoes such as those in the Culex pipiens complex are important vectors of disease. This study was conducted to genetically characterize Cx. pipiens complex populations in the state of Colorado, USA, and to determine the number of genetic clusters represented by the data. Thirteen populations located among four major river basins were sampled (n = 597 individuals) using a panel of 14 microsatellites. The lowest-elevation sites had the highest Expected Heterozygosity (HE) values (range 0.54–0.65). AMOVA results indicated the presence of statistically significant amounts of variation within each level when populations were analyzed as one group or when they were grouped either by river basin or by their position on the east or west side of the Rocky Mountains. Most pairwise FST values were significant via permutation test (range 0–0.10), with the highest values from comparisons with Lamar, in southeast CO. A neighbor joining tree based on Cavalli–Sforza and Edwards’s chord distances was consistent with the geographic locations of populations, as well as with the AMOVA results. There was a significant isolation by distance effect, and the cluster analysis resolved five groups. Individuals were also assayed with an additional microsatellite marker, Cxpq78, proposed to be monomorphic in Cx. pipiens but polymorphic in the closely related but biologically distinct species Cx. quinquefasciatus. Low frequencies (≤3%) of Cx. quinquefasciatus alleles for this marker were noted, and mostly confined to populations along the Interstate 25 corridor. Pueblo was distinct in that it had 10% Cx. quinquefasciatus alleles, mostly of one allele size. The degree of population genetic structure observed in this study is in contrast with that of Cx. tarsalis, the other major vector of WNV in the western U.S., and likely reflects the two species’ different dispersal strategies. PMID:23094068

  18. Characterization of Culex pipiens complex (Diptera: Culicidae) populations in Colorado, USA using microsatellites.

    PubMed

    Kothera, Linda; Godsey, Marvin S; Doyle, Michael S; Savage, Harry M

    2012-01-01

    Mosquitoes such as those in the Culex pipiens complex are important vectors of disease. This study was conducted to genetically characterize Cx. pipiens complex populations in the state of Colorado, USA, and to determine the number of genetic clusters represented by the data. Thirteen populations located among four major river basins were sampled (n = 597 individuals) using a panel of 14 microsatellites. The lowest-elevation sites had the highest Expected Heterozygosity (H(E)) values (range 0.54-0.65). AMOVA results indicated the presence of statistically significant amounts of variation within each level when populations were analyzed as one group or when they were grouped either by river basin or by their position on the east or west side of the Rocky Mountains. Most pairwise F(ST) values were significant via permutation test (range 0-0.10), with the highest values from comparisons with Lamar, in southeast CO. A neighbor joining tree based on Cavalli-Sforza and Edwards's chord distances was consistent with the geographic locations of populations, as well as with the AMOVA results. There was a significant isolation by distance effect, and the cluster analysis resolved five groups. Individuals were also assayed with an additional microsatellite marker, Cxpq78, proposed to be monomorphic in Cx. pipiens but polymorphic in the closely related but biologically distinct species Cx. quinquefasciatus. Low frequencies (≤3%) of Cx. quinquefasciatus alleles for this marker were noted, and mostly confined to populations along the Interstate 25 corridor. Pueblo was distinct in that it had 10% Cx. quinquefasciatus alleles, mostly of one allele size. The degree of population genetic structure observed in this study is in contrast with that of Cx. tarsalis, the other major vector of WNV in the western U.S., and likely reflects the two species' different dispersal strategies.

  19. Many compatible Wolbachia strains coexist within natural populations of Culex pipiens mosquito

    PubMed Central

    Duron, O; Raymond, M; Weill, M

    2011-01-01

    Maternally inherited Wolbachia often manipulate the reproduction of arthropods to promote their transmission. In most species, Wolbachia exert a form of conditional sterility termed cytoplasmic incompatibility (CI), characterized by the death of embryos produced by the mating between individuals with incompatible Wolbachia infections. From a theoretical perspective, no stable coexistence of incompatible Wolbachia infections is expected within host populations and CI should induce the invasion of one strain or of a set of compatible strains. In this study, we investigated this prediction on CI dynamics in natural populations of the common house mosquito Culex pipiens. We surveyed the Wolbachia diversity and the expression of CI in breeding sites of the south of France between 1990 and 2005. We found that geographically close C. pipiens populations harbor considerable Wolbachia diversity, which is stably maintained over 15 years. We also observed a very low frequency of infertile clutches within each sampled site. Meanwhile, mating choice experiments conducted in laboratory conditions showed that assortative mating does not occur. Overall, this suggests that a large set of compatible Wolbachia strains are always locally dominant within mosquito populations thus, fitting with the theoretical expectations on CI dynamics. PMID:21119702

  20. 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...

  1. 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...

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

    PubMed

    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.

  3. Predicting Culex pipiens/restuans population dynamics by interval lagged weather data

    PubMed Central

    2013-01-01

    Background Culex pipiens/restuans mosquitoes are important vectors for a variety of arthropod borne viral infections. In this study, the associations between 20 years of mosquito capture data and the time lagged environmental quantities daytime length, temperature, precipitation, relative humidity and wind speed were used to generate a predictive model for the population dynamics of this vector species. Methods Mosquito population in the study area was represented by averaged time series of mosquitos counts captured at 6 sites in Cook County (Illinois, USA). Cross-correlation maps (CCMs) were compiled to investigate the association between mosquito abundances and environmental quantities. The results obtained from the CCMs were incorporated into a Poisson regression to generate a predictive model. To optimize the predictive model the time lags obtained from the CCMs were adjusted using a genetic algorithm. Results CCMs for weekly data showed a highly positive correlation of mosquito abundances with daytime length 4 to 5 weeks prior to capture (quantified by a Spearman rank order correlation of rS = 0.898) and with temperature during 2 weeks prior to capture (rS = 0.870). Maximal negative correlations were found for wind speed averaged over 3 week prior to capture (rS = −0.621). Cx. pipiens/restuans population dynamics was predicted by integrating the CCM results in Poisson regression models. They were used to simulate the average seasonal cycle of the mosquito abundance. Verification with observations resulted in a correlation of rS = 0.899 for daily and rS = 0.917 for weekly data. Applying the optimized models to the entire 20-years time series also resulted in a suitable fit with rS = 0.876 for daily and rS = 0.899 for weekly data. Conclusions The study demonstrates the application of interval lagged weather data to predict mosquito abundances with a feasible accuracy, especially when related to weekly Cx. pipiens

  4. 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

  5. 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.

  6. 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

  7. Naturally occurring incompatibilities between different Culex pipiens pallens populations as the basis of potential mosquito control measures.

    PubMed

    Chen, Lin; Zhu, Changliang; Zhang, Donghui

    2013-01-01

    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. 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. 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 supplement to other control measures.

  8. 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

  9. Effects of latitude and longitude on the population structure of Culex pipiens s.l., vectors of West Nile virus in North America.

    PubMed

    Edillo, Frances; Kiszewski, Anthony; Manjourides, Justin; Pagano, Marcello; Hutchinson, Michael; Kyle, Andrew; Arias, Jorge; Gaines, David; Lampman, Richard; Novak, Robert; Foppa, Ivo; Lubelcyzk, Charles; Smith, Robert; Moncayo, Abelardo; Spielman, Andrew

    2009-11-01

    We assessed the structure and latitudinal selection that might result in sensitivities to critical day-lengths that trigger diapause between Culex pipiens populations distributed along North-South and East-West axes in eastern North America. Strong population structure between Cx. p. pipiens and Cx. p. quinquefasciatus existed. Among Cx. p. pipiens, a 100-km increase in the latitudinal change resulted in an increased square root of F(ST) by 0.002. A 100-km increase in the longitudinal change caused an increased square root of F(ST) by 0.035. A lack of latitudinal influence on the structure between Cx. p. pipiens populations suggests a uniform signal using the 12 microsatellite markers, which might increase the risk of West Nile virus (WNV) transmission toward northern areas because of longer breeding season, extend host-seeking period, and larger population size. Northern Cx. p. pipiens may have undergone additional generations before diapause is triggered, magnifying population size when WNV amplification is peaking.

  10. Effects of Latitude and Longitude on the Population Structure of Culex pipiens s.l., Vectors of West Nile Virus in North America

    PubMed Central

    Edillo, Frances; Kiszewski, Anthony; Manjourides, Justin; Pagano, Marcello; Hutchinson, Michael; Kyle, Andrew; Arias, Jorge; Gaines, David; Lampman, Richard; Novak, Robert; Foppa, Ivo; Lubelcyzk, Charles; Smith, Robert; Moncayo, Abelardo

    2010-01-01

    We assessed the structure and latitudinal selection that might result in sensitivities to critical day-lengths that trigger diapause between Culex pipiens populations distributed along North-South and East-West axes in eastern North America. Strong population structure between Cx. p. pipiens and Cx. p. quinquefasciatus existed. Among Cx. p. pipiens, a 100-km increase in the latitudinal change resulted in an increased square root of FST by 0.002. A 100-km increase in the longitudinal change caused an increased square root of FST by 0.035. A lack of latitudinal influence on the structure between Cx. p. pipiens populations suggests a uniform signal using the 12 microsatellite markers, which might increase the risk of West Nile virus (WNV) transmission toward northern areas because of longer breeding season, extend host-seeking period, and larger population size. Northern Cx. p. pipiens may have undergone additional generations before diapause is triggered, magnifying population size when WNV amplification is peaking. PMID:19861620

  11. Breeding phenology in Rana temporaria. Local variation is due to pond temperature and population size.

    PubMed

    Loman, Jon

    2016-09-01

    Frog breeding phenology in temperate zones is usually compared to progress of spring temperatures at a regional scale. However, local populations may differ substantially in phenology. To understand this, local climate and other aspects must be studied. In this study, breeding phenology of the common frog, Rana temporaria, in a set of ponds in southern Sweden is analyzed. There was within year a variation of up to 3 weeks in start of breeding among local populations. Water temperature was measured in the ponds, and breeding tended to be earlier in warmer ponds (surprise!). Breeding was also earlier in ponds with a large breeding congregation. Alternative reasons for these patterns are suggested and discussed. There was a large residual variation. The common frog has a wide range of acceptable wintering sites, and I hypothesize that the particular choice by a local population may explain part of this residual variation.

  12. Population structure and distribution patterns of the sibling mosquito species Culex pipiens and Culex torrentium (Diptera: Culicidae) reveal different evolutionary paths.

    PubMed

    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. 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

  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. High prevalence and lineage diversity of avian malaria in wild populations of great tits (Parus major) and mosquitoes (Culex pipiens).

    PubMed

    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.

  17. 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.

  18. Population differentiation in G matrix structure due to natural selection in Rana temporaria.

    PubMed

    Cano, José Manuel; Laurila, Anssi; Pało, Jukka; Merilä, Juha

    2004-09-01

    The additive genetic variance-covariance matrix (G) is a concept central to discussions about evolutionary change over time in a suite of traits. However, at the moment we do not know how fast G itself changes as a consequence of selection or how sensitive it is to environmental influences. We investigated possible evolutionary divergence and environmental influences on G using data from a factorial common-garden experiment where common frog (Rana temporaria) tadpoles from two divergent populations were exposed to three different environmental treatments. G-matrices were estimated using an animal model approach applied to data from a NCII breeding design. Matrix comparisons using both Flury and multivariate analysis of variance methods revealed significant differences in G matrices both between populations and between treatments within populations, the former being generally larger than the latter. Comparison of levels of population differentiation in trait means using Q(ST) indices with that observed in microsatellite markers (F(ST)) revealed that the former values generally exceeded the neutral expectation set by F(ST). Hence, the results suggest that intraspecific divergence in G matrix structure has occurred mainly due to natural selection.

  19. Proximate causes of adaptive growth rates: growth efficiency variation among latitudinal populations of Rana temporaria.

    PubMed

    Lindgren, B; Laurila, A

    2005-07-01

    In ectothermic organisms, declining season length and lower temperature towards higher latitudes often select for latitudinal variation in growth and development. However, the energetic mechanisms underlying this adaptive variation are largely unknown. We investigated growth, food intake and growth efficiency of Rana temporaria tadpoles from eight populations along a 1500 km latitudinal gradient across Sweden. To gain an insight into the mechanisms of adaptation at organ level, we also examined variation in tadpole gut length. The tadpoles were raised at two temperatures (16 and 20 degrees C) in a laboratory common garden experiment. We found increased growth rate towards higher latitudes, regardless of temperature treatment. This increase in growth was not because of a higher food intake rate, but populations from higher latitudes had higher growth efficiency, i.e. they were more efficient at converting ingested food into body mass. Low temperature reduced growth efficiency most strongly in southern populations. Relative gut length increased with latitude, and tadpoles at low temperature tended to have longer guts. However, variation in gut length was not the sole adaptive explanation for increased growth efficiency as latitude and body length still explained significant amounts of variation in growth efficiency. Hence, additional energetic adaptations are probably involved in growth efficiency variation along the latitudinal gradient.

  20. 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.

  1. Within-population polymorphism of sex-determination systems in the common frog (Rana temporaria).

    PubMed

    Rodrigues, N; Betto-Colliard, C; Jourdan-Pineau, H; Perrin, N

    2013-07-01

    In sharp contrast with birds and mammals, the sex chromosomes of ectothermic vertebrates are often undifferentiated, for reasons that remain debated. A linkage map was recently published for Rana temporaria (Linnaeus, 1758) from Fennoscandia (Eastern European lineage), with a proposed sex-determining role for linkage group 2 (LG2). We analysed linkage patterns in lowland and highland populations from Switzerland (Western European lineage), with special focus on LG2. Sibship analyses showed large differences from the Fennoscandian map in terms of recombination rates and loci order, pointing to large-scale inversions or translocations. All linkage groups displayed extreme heterochiasmy (total map length was 12.2 cM in males, versus 869.8 cM in females). Sex determination was polymorphic within populations: a majority of families (with equal sex ratios) showed a strong correlation between offspring phenotypic sex and LG2 paternal haplotypes, whereas other families (some of which with female-biased sex ratios) did not show any correlation. The factors determining sex in the latter could not be identified. This coexistence of several sex-determination systems should induce frequent recombination of X and Y haplotypes, even in the absence of male recombination. Accordingly, we found no sex differences in allelic frequencies on LG2 markers among wild-caught male and female adults, except in one high-altitude population, where nonrecombinant Y haplotypes suggest sex to be entirely determined by LG2. Multifactorial sex determination certainly contributes to the lack of sex-chromosome differentiation in amphibians. © 2013 The Authors. Journal of Evolutionary Biology © 2013 European Society For Evolutionary Biology.

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

    PubMed

    Mulatti, Paolo; Ferguson, Heather M; Bonfanti, Lebana; Montarsi, Fabrizio; Capelli, Gioia; Marangon, Stefano

    2014-01-15

    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. 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 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. 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.

  3. 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

  4. Dynamics of testis-ova in a wild population of Japanese pond frogs, Rana nigromaculata.

    PubMed

    Kobayashi, Tohru; Kumakura, Masahiko; Yoshie, Sumio; Sugishima, Tomomi; Horie, Yoshifumi

    2015-02-01

    Although many studies have reported the occurrence of testis-ova in wild frog populations, the origin and trigger of testis-ova differentiation/development remain unclear. A high frequency of testis-ova has been previously reported for wild populations of the Japanese pond frog, Rana nigromaculata (cf. Iwasawa and Asai, '59). In the present study, we aimed to clarify the dynamics of testis-ova in this frog species, including the origin and artificial induction of testis-ova. Testis-ova were observed in both mature frogs and puberty-stage frogs (i.e., 0- and 1-year-old frogs). However, the early stages of testis-ova (~pachytene stage) were mostly observed in puberty-stage male frogs at the onset of spermatogenesis. The early stages of testis-ova were observed in the cysts of early secondary spermatogonia and the single cysts of the primary spermatogonium. This finding indicates that testis-ova differentiation occurs during spermatogonial proliferation and that it is correlated with the initiation of spermatogenesis. We also examined whether estrogen exposure induced testis-ova differentiation and how it is correlated with the progression of spermatogenesis. When 1-year-old frogs were exposed to estradiol-17β during spring (i.e., when spermatogenesis was initiated), testis-ova differentiation was induced in a dose-dependent manner. However, this phenomenon did not occur in 1-year-old frogs during summer, (i.e., when the transition from spermatogonia to spermatocytes mainly occurs). These results present the first evidence that testis-ova of the Japanese pond frog are derived from primary and early secondary spermatogonia, and that estrogen exposure induces testis-ova differentiation accompanied by the initiation of spermatogenesis.

  5. Transcriptomics of differential vector competence: West Nile virus infection in two populations of Culex pipiens quinquefasciatus linked to ovary development.

    PubMed

    Shin, Dongyoung; Civana, Ayse; Acevedo, Carolina; Smartt, Chelsea T

    2014-06-22

    Understanding mechanisms that contribute to viral dissemination in mosquito vectors will contribute to our ability to interfere with the transmission of viral pathogens that impact public health. The expression of genes in two Culex pipiens quinquefasciatus populations from Florida with known differences in vector competence to West Nile virus (WNV) were compared using high throughput sequencing. A total of 15,176 transcripts were combined for comparison of expression differences between the two populations and 118 transcripts were differentially expressed (p < 0.05). The fold change in expression of the differentially expressed genes ranged from -7.5 - 6.13. The more competent population for WNV (Gainesville) over expressed 77 genes and down regulated 44 genes, compared with the less competent population for WNV (Vero Beach). Also, splicing analysis identified 3 transcripts with significantly different splice forms between the two populations. The functional analysis showed that the largest proportion of transcripts was included in the catalytic activity and transporter activity groups except for those in the unknown group. Interestingly, the up- regulated gene set contained most of the catalytic activity function and the down- regulated gene set had a notable proportion of transcripts with transporter activity function. Immune response category was shown in only the down regulated gene set, although those represent a relatively small portion of the function. Several different vitellogenin genes were expressed differentially. Based on the RNAseq data analysis, ovary development was compared across the populations and following WNV infection. There were significant differences among the compared groups. This study suggests that ovary development is correlated to vector competence in two Culex populations in Florida. Both populations control energy allocations to reproduction as a response to WNV. This result provides novel insight into the defense mechanism used by

  6. The emerging amphibian pathogen Batrachochytrium dendrobatidis globally infects introduced populations of the North American bullfrog, Rana catesbeiana

    PubMed Central

    Garner, Trenton W.J; Perkins, Matthew W; Govindarajulu, Purnima; Seglie, Daniele; Walker, Susan; Cunningham, Andrew A; Fisher, Matthew C

    2006-01-01

    Batrachochytrium dendrobatidis is the chytridiomycete fungus which has been implicated in global amphibian declines and numerous species extinctions. Here, we show that introduced North American bullfrogs (Rana catesbeiana) consistently carry this emerging pathogenic fungus. We detected infections by this fungus on introduced bullfrogs from seven of eight countries using both PCR and microscopic techniques. Only native bullfrogs from eastern Canada and introduced bullfrogs from Japan showed no sign of infection. The bullfrog is the most commonly farmed amphibian, and escapes and subsequent establishment of feral populations regularly occur. These factors taken together with our study suggest that the global threat of B. dendrobatidis disease transmission posed by bullfrogs is significant. PMID:17148429

  7. Population genetic patterns suggest a behavioural change in wild common frogs (Rana temporaria) following disease outbreaks (Ranavirus).

    PubMed

    Teacher, Amber G F; Garner, Trenton W J; Nichols, Richard A

    2009-08-01

    We use 14 microsatellite loci to investigate the impact of a viral disease (Ranavirus) on the population genetic structure of wild common frogs (Rana temporaria). Populations with a history of Ranavirus mortalities (and 83% declines in the number of frogs) were compared with populations with no history of infection. Infected ponds showed significantly elevated F(IS) (homozygote excess), significantly reduced relatedness, and no detectable effect on allelic richness. We hypothesize that the elevated F(IS) and reduced relatedness are consequences of assortative mating, and that allelic richness is maintained by immigration from nearby populations. Simulations indicate that the elevated F(IS) cannot be explained by population size reductions, but can indeed be explained by assortative mating (even if a mate choice locus is unlinked to the genetic markers). While the majority of studies consider demographic outcomes following disease outbreaks, our results indicate that emerging infectious diseases could also result in behavioural changes.

  8. Recognition of a Novel Melanotic Mutant in a Field Population of Culex Pipiens Quinquefasciatus in Southern Zambia

    PubMed Central

    Kent, Rebekah J.; Mharakurwa, Sungano; Hamapumbu, Harry; Norris, Douglas E.

    2014-01-01

    Genetic mutations controlling eye color, fat body color, structural abnormalities, and insecticide resistance are common in mosquitoes. We have identified a novel color variant of Culex pipiens quinquefasciatus characterized by a heavily pigmented integument in adult specimens circulating in field populations of this species in southern Zambia. Mosquitoes were collected monthly by pyrethrum spray catch between November and May 2004–05 and 2005–06, with between 25% and 80% of the total Cx. p. quinquefasciatus collections comprising this pigmented variety. The identity of pigmented specimens was morphologically confirmed as Cx. p. quinquefasciatus by examination of orientation of the dorsal and ventral arms of the male genitalia of F1 male progeny reared in the laboratory, and molecularly verified by diagnostic polymerase chain reaction and nicotinamide adenine dinucleotide phosphate dehydrogenase subunit 4 nucleotide sequence homology. Preliminary laboratory rearings indicated that the pigmentation was heritable and not influenced by larval habitat and environmental conditions. Further investigation into the mechanism of the mutation, inheritance patterns, and potential linkage to additional markers is pending. PMID:17536371

  9. Amphibian population genetics in agricultural landscapes: does viniculture drive the population structuring of the European common frog (Rana temporaria)?

    PubMed

    Lenhardt, Patrick P; Brühl, Carsten A; Leeb, Christoph; Theissinger, Kathrin

    2017-01-01

    Amphibian populations have been declining globally over the past decades. The intensification of agriculture, habitat loss, fragmentation of populations and toxic substances in the environment are considered as driving factors for this decline. Today, about 50% of the area of Germany is used for agriculture and is inhabited by a diverse variety of 20 amphibian species. Of these, 19 are exhibiting declining populations. Due to the protection status of native amphibian species, it is important to evaluate the effect of land use and associated stressors (such as road mortality and pesticide toxicity) on the genetic population structure of amphibians in agricultural landscapes. We investigated the effects of viniculture on the genetic differentiation of European common frog (Rana temporaria) populations in Southern Palatinate (Germany). We analyzed microsatellite data of ten loci from ten breeding pond populations located within viniculture landscape and in the adjacent forest block and compared these results with a previously developed landscape permeability model. We tested for significant correlation of genetic population differentiation and landscape elements, including land use as well as roads and their associated traffic intensity, to explain the genetic structure in the study area. Genetic differentiation among forest populations was significantly lower (median pairwise FST = 0.0041 at 5.39 km to 0.0159 at 9.40 km distance) than between viniculture populations (median pairwise FST = 0.0215 at 2.34 km to 0.0987 at 2.39 km distance). Our analyses rejected isolation by distance based on roads and associated traffic intensity as the sole explanation of the genetic differentiation and suggest that the viniculture landscape has to be considered as a limiting barrier for R. temporaria migration, partially confirming the isolation of breeding ponds predicted by the landscape permeability model. Therefore, arable land may act as a sink habitat, inhibiting genetic

  10. Turning population trend monitoring into active conservation: Can we save the Cascades Frog (Rana cascadae) in the Lassen region of California?

    Treesearch

    Gary M. Fellers; Karen L. Pope; Jonathan E. Stead; Michelle S. Koo; Hartwell H. Welsh

    2007-01-01

    Monitoring the distribution, population size, and trends of declining species is necessary to evaluate their vulnerability to extinction. It is the responsibility of scientists to alert management professionals of the need for preemptive action if a species approaches imminent, regional extirpation. This is the case with Rana cascadae (Cascades Frog...

  11. 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...

  12. 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...

  13. A new species of Rhabdias from lungs of the wood frog, Rana sylvatica, in North America: the last sibling of Rhabdias ranae?

    PubMed

    Tkach, Vasyl V; Kuzmin, Yuriy; Pulis, Eric E

    2006-06-01

    Rhabdias bakeri n. sp. is described from specimens found in lungs of the wood frog, Rana sylvatica, from North Dakota. The new species has previously been mistakenly identified as Rhabdias ranae Walton, 1929, a common parasite of the leopard frog, Rana pipiens. The new species differs from R. ranae and Rhabdias joaquinensis Ingles, 1935 by the shape and size of pseudolabia, shape and size of buccal capsule, and wider esophageal bulb. Molecular analysis based on the partial sequences of nuclear 18S rDNA gene, complete sequences of internal transcribed spacer region, and partial sequences of 28S gene demonstrates clear differences between Rhabdias from Ra. sylvatica and Ra. pipiens, and supports the status of R. bakeri as a new species.

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

    PubMed Central

    KNAPP, Roland A.; BOIANO, Daniel M.; VREDENBURG, Vance T.

    2007-01-01

    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

  15. 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.

  16. Insecticide resistance alleles in wetland and residential populations of the West Nile virus vector Culex pipiens in New Jersey.

    PubMed

    Johnson, Brian J; Fonseca, Dina M

    2016-03-01

    In spite of the extensive use of insecticides to control Culex pipiens in the aftermath of West Nile virus, knowledge of the spatial distribution and frequency of insecticide resistance in this species is poorly understood in the United States. This paper reports on the occurrence of upregulated esterases that detoxify organophosphates (OPs) and mutations conferring resistance to pyrethroid insecticides in natural and developed areas of New Jersey. We report the first observations of the OP resistance alleles Ester(B1) and Ester(2) and the classical knockdown resistance (kdr) mutation L1014F in New Jersey Cx. pipiens. Upregulated Ester(B1) peaked at 23% (mean ± SE = 12 ± 2.3%) and Ester(2) at 14% (8 ± 1.8%), and both were widely distributed. L1014F, which confers strong resistance to pyrethroids when homozygous, was also widely distributed and ranged in frequency from 2 to 19% (5.1% heterozygous individuals and 1.4% homozygous). We have demonstrated that OP resistance is common and broadly distributed in New Jersey Cx. pipiens, and that homozygous individuals resistant to pyrethroids are present. Further, we have detected double mutants at Ester and kdr, a condition that may annul the purging effects of insecticide rotations. Our results therefore indicate the need for continued monitoring of insecticide resistance in order to achieve effective mosquito control. © 2015 Society of Chemical Industry.

  17. 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

  18. 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.

  19. Growth, size and age at maturity of the agile frog (Rana dalmatina) in an Iberian Peninsula population.

    PubMed

    Sarasola-Puente, Vanessa; Gosá, Alberto; Oromí, Neus; Madeira, María José; Lizana, Miguel

    2011-06-01

    The mean age of a population of agile frogs (Rana dalmatina) from the Iberian Peninsula was estimated using mark and recapture and skeletochronology. Life-history parameters, including growth rate, body length, age and size at maturity, sexual dimorphism and longevity, were studied. The regression between age and snout-vent length (SVL) was highly significant in both sexes. Males reached sexual maturity at two years of age, although sometimes they can reach it at only one year of age. The average SVL at maturity was 51.75 mm (standard error (SE)=0.71; n=45). Females reached sexual maturity at two years of age with an average SVL of 62.14 mm (SE=2.20; n=14). A subset of the female population reached sexual maturity at three years of age. Growth was rapid until sexual maturity was reached. There was an overlap of SVL between different age classes. Growth was continuous, fulfilling the conditions of Von Bertalanffy's model. The growth coefficient (K) was 0.840 in males and 0.625 in females. The maximum SVL was greater in females (73.00 mm) than in males (59.50mm). Sexual dimorphism was significantly biased towards females in all age classes. The maximum longevity observed was 6 years in females and 8 years in males. Management strategies for agile frogs should take into account factors such as these life-history characteristics.

  20. Testing the role of phenotypic plasticity for local adaptation: growth and development in time-constrained Rana temporaria populations.

    PubMed

    Lind, M I; Johansson, F

    2011-12-01

    Phenotypic plasticity can be important for local adaptation, because it enables individuals to survive in a novel environment until genetic changes have been accumulated by genetic accommodation. By analysing the relationship between development rate and growth rate, it can be determined whether plasticity in life-history traits is caused by changed physiology or behaviour. We extended this to examine whether plasticity had been aiding local adaptation, by investigating whether the plastic response had been fixed in locally adapted populations. Tadpoles from island populations of Rana temporaria, locally adapted to different pool-drying regimes, were monitored in a common garden. Individual differences in development rate were caused by different foraging efficiency. However, developmental plasticity was physiologically mediated by trading off growth against development rate. Surprisingly, plasticity has not aided local adaptation to time-stressed environments, because local adaptation was not caused by genetic assimilation but on selection on the standing genetic variation in development time. © 2011 The Authors. Journal of Evolutionary Biology © 2011 European Society For Evolutionary Biology.

  1. Discordant patterns of nuclear and mitochondrial introgression in Iberian populations of the European common frog (Rana temporaria).

    PubMed

    Veith, Michael; Baumgart, Andrea; Dubois, Alain; Ohler, Annemarie; Galán, Pedro; Vieites, David R; Nieto-Román, Sandra; Vences, Miguel

    2012-01-01

    Amphibians often show complex histories of intraspecific and interspecific genetic introgression, which might differ in mitochondrial and nuclear genes. In our study of the genetic differentiation of the European common frog, Rana temporaria (159 specimens from 23 populations were analyzed for 24 presumptive allozyme loci; 82 specimens were sequenced for a 540-bp fragment of the mitochondrial 16S rRNA gene), multilocus correspondence analysis (CA) and Bayesian assignment tests of the nuclear data were concordant in identifying 2 population groups corresponding to 1) the Pyrenees in the east and 2) the Galicia and Asturias regions in the west, the latter corresponding to the subspecies R. temporaria parvipalmata. Geographically intermediate populations were genetically intermediate in the allozyme CA and, less clearly in the Bayesian assignment, with mitochondrial haplotypes exclusively belonging to the parvipalmata group. This indicates different degrees of introgression in the mitochondrial and nuclear genomes. Although Pyrenean high-altitude populations are morphologically distinct from low-altitude populations, these 2 groups were not separate clusters in any analysis. This suggests that the morphological differences may be due to fast adaptation to elevational gradients, likely under maintenance of gene flow, and that the underlying genetic changes are not detectable by the analyzed markers. We argue that a parsimonious explanation for the observed pattern along the east-west axis in northern Spain may be competition between invading and resident populations, with no need to invoke selection. However, in order to conclusively rule out selective processes, additional and finer scale data are required to test for asymmetric mating preference/behaviour, sex-biased gene flow, or sex-biased survival of potential hybrids.

  2. 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.

  3. Insecticide resistance and cytochrome-P450 activation in unfed and blood-fed laboratory and field populations of Culex pipiens pallens.

    PubMed

    Chang, Kyu-Sik; Kim, Heung-Chul; Klein, Terry A; Ju, Young Ran

    2017-01-01

    Understanding the mechanisms of insecticide resistance to vector mosquitoes is critical for the implementation of effective control measures. A nulliparous susceptible Culex pipiens pallens (KSCP) laboratory colony and two field strains from Paju (PAJ) and Jeonju (JEO) Korea were evaluated for susceptibility to five pesticides by microapplication techniques. Unfed PAJ and JEO females demonstrated increased resistance compared to unfed KSCP females, respectively. While blood-fed KSCP females demonstrated <10-fold decreased susceptibility to pesticides compared to unfed KSCP females, blood-fed PAJ and JEO females demonstrated 25.0-50.0- and 16.0-38.6-fold increased resistance compared to unfed PAJ and JEO females, respectively. Unfed and blood-fed groups were assayed for α- and β-esterase, glutathione S-transferases, and cytochrome P-450 (P450) enzyme activity assays. P450 activity was 58.8- and 72.8-fold higher for unfed PAJ and JEO females, respectively, than unfed KSCP females. P450 enzyme activity of KSCP females assayed 1 and 7 days after a blood meal increased by 14.5- and 11.8-fold, respectively, compared to unfed KSCP females, while PAJ and JEO females demonstrated 164.9- and 148.5- and 170.7- and 160.4-fold increased activity, respectively, compared to unfed females of each population. However, other three resistance-related metabolic enzymes showed low activation at <10-fold after a blood meal. The data demonstrate that P450 acts on elevated insecticide resistance after blood meals in resistant field populations. Our findings might reveal that suppressing of the P450 protein by artificial gene mutation increases insecticidal susceptibility of Cx. pipiens and will promise effective vector mosquito control.

  4. 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

  5. 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...

  6. 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.

  7. Influence of warming tendency on Culex pipiens population abundance and on the probability of West Nile fever outbreaks (Israeli Case Study: 2001-2005).

    PubMed

    Paz, Shlomit; Albersheim, Iris

    2008-03-01

    Climate change and West Nile fever (WNV) are both subjects of global importance. Many mosquitoes and the diseases they carry, including West Nile virus (WNV), are sensitive to temperature increase. The current study analyzes the lag correlations between weather conditions (especially air temperature) and 1) Culex pipiens mosquito population abundance, and 2) WNF frequency in humans, between 2001 and 2005 in Israel. These 5 years follow a long period with a documented tendency for temperature increase in the hot season in the country. Monthly anomalies of minimum and maximum temperatures, relative seasonal rainfall contribution, mosquito samplings (hazard level), and WNF cases (hospital admission dates and patients' addresses) were analyzed. Logistic regression was calculated between the climatic data and the mosquito samples, as Spearman correlations and Pearson cross-correlations were calculated between daily temperature values (or daily precipitation amounts) and the hospital admission dates. It was found that the disease appearance reflects the population distribution, while the risk tends to escalate around the metropolis characterized by an urban heat island. Positive anomalies of the temperature during the study period appear to have facilitated the mosquito abundance and, consequently, the disease emergence in humans. An important finding is the potential influence of extreme heat in the early spring on the vector population increase and on the disease's appearance weeks later. Awareness of such situations at the beginning of the spring may help authorities to reduce the disease risk before it becomes a real danger.

  8. [Role of the mosquitoes Culex pipienis f. pipiens and Cx. pipiens f. molestus (Diptera, Culicidae) in the spread of West Nile virus in the south of Russia].

    PubMed

    Fedorova, M V; Shaĭkevich, E V

    2013-01-01

    Cx. pipiens is one of the major vectors of West Nile virus in the south of Russia and it is represented by the autogenic form, molestus, and the non-autogenic form, pipiens. The spatial distribution of its larvae and food preferences of females was investigated to assess the potential role of each form in the spread of the virus. The taxonomic status of the mosquitoes was determined from their capacity for autogenicity (543 specimens) and the type of mitochondrial DNA (348 specimens). The mosquitoes of the pipiens form were shown to develop in the open urban and rural reservoirs and the females were non-autogenic. Cx. pipiens form molestus was found only in the urban biotopes, mainly in the flooded basements and, in some years, also in the open reservoirs where it formed a mixed population with Cx. pipiens form pipiens. Both autogenic (85-95.2%) and non-autogenic specimens were identified among the females with the mitotype in form molestus. Genetic analysis of Cx. pipiens females collected under the Berezantsev bell and in the trap with a bird showed that both forms were attracted to man and the bird. The findings suggest that Cx. pipiens form pipiens can transmit West Nile virus to humans both in the town and in its suburb and Cx. pipiens form molestus can only in urban areas.

  9. Emergence of Culex pipiens from overwintering hibernacula.

    PubMed

    Ciota, Alexander T; Drummond, Cori L; Drobnack, Jason; Ruby, Meghan A; Kramer, Laura D; Ebel, Gregory D

    2011-03-01

    Overwintering populations of Culex pipiens, the principal enzootic vector of West Nile virus in the northeastern USA, were studied over 3 consecutive winters from 2006 to 2008, using mark-recapture techniques to determine when Cx. pipiens females began to disperse from overwintering hibernacula and how their survival influenced early season populations. In February of each year, Cx. pipiens were aspirated and marked using fluorescent powder; 4,067, 752, and 3,070 diapausing Cx. pipiens were marked in each successive year. Mosquitoes were then trapped from mid-April to early May of each year using 19 Centers for Disease Control and Prevention (CDC) light traps and 16 CDC gravid traps. A total of 348, 39, and 111 Culex mosquitoes were captured in the spring of 2006, 2007, and 2008, respectively. The number of mosquitoes marked in overwintering habitats is generally positively correlated with the number of mosquitoes recaptured in the early spring (linear regression, R2 = 0.79, P = 0.04), yet results also suggest that seasonal variations beyond overwintering population size are likely important in determining the success of emergent populations. A single marked Cx. pipiens was captured in both 2006 and 2008. In 2006, the mosquito was captured 0.5 km from its overwintering site while in 2008 the mosquito was captured 0.3 km from its overwintering site. In all study years, mosquitoes consistently began exiting overwintering hibernacula the 3rd week of April, yet evidence of earlier exodus was observed in 2007, when outside temperatures were significantly higher in preceding days and months.

  10. 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

  11. THE ORIGIN OF PROTEIN AND FATTY YOLK IN RANA PIPIENS

    PubMed Central

    Ward, Robert T.

    1962-01-01

    Electron microscope studies of young oocytes have demonstrated that the plate-like, hexagonally shaped yolk bodies previously observed in living cells are wholly within the substance of oocyte mitochondria and that they remain within these mitochondria while increasing in size. These bodies possess a crystalline structure consisting of what appear to be lines, with a spacing of 70 to 85 A, and appear very dense in the electron microscope. After formalin fixation such bodies give an intense positive test for protein, and when viewed in the electron microscope are only slightly less dense than after OsO4 fixation. Evidence is presented for the origin of these crystals within a single crista. The clusters of yolk globules previously studied in living cells are seen to consist of several types of bodies, but an irregular dense droplet predominates. This dense material is apparently secreted by small spherical bodies which, the evidence suggests, originate from the breaking up of filamentous mitochondria and which possess an outer double membrane and sometimes internal cristalike membranes. When thin sections of young oocytes are immersed in xylol the dense globules of the clusters are dissolved, but the hexagonal bodies are unaffected, indicating that the globules are of a predominantly fatty nature, while the hexagonal bodies are of a predominantly protein nature. Examination of mature or almost mature oocytes has revealed that the main body of the yolk platelets is crystalline in nature and is surrounded by a thick matrix which, in light microscope study, masks the fact that the face view of the main body of the platelets is often hexagonal. The spacing within the main body is found to be 70 to 85 A. The crystal laminae of this material can be resolved quite clearly into rows of particles. Dense globules of varying sizes are found in the cytoplasm between the platelets. When thin sections of these OsO4-fixed oocytes are immersed in xylol, the material of the globules is extracted and the crystalline material of the platelets remains unaffected, indicating the fatty nature of the globules and the protein nature of the platelets. The platelets of the mature egg resemble the hexagon bodies, previously described in young oocytes, in their protein nature, their crystalline spacing, and their hexagonal outline. This is given as strong evidence for the origin of the mature platelets by the growth of the intramitochondrial hexagon bodies. The biochemical implications of this study are discussed. PMID:14004944

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

    PubMed Central

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

    2016-01-01

    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

  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.

  15. Big mountains but small barriers: Population genetic structure of the Chinese wood frog (Rana chensinensis) in the Tsinling and Daba Mountain region of northern China

    PubMed Central

    Zhan, Aibin; Li, Cheng; Fu, Jinzhong

    2009-01-01

    Background Amphibians in general are poor dispersers and highly philopatric, and landscape features often have important impacts on their population genetic structure and dispersal patterns. Numerous studies have suggested that genetic differentiation among amphibian populations are particularly pronounced for populations separated by mountain ridges. The Tsinling Mountain range of northern China is a major mountain chain that forms the boundary between the Oriental and Palearctic zoogeographic realms. We studied the population structure of the Chinese wood frog (Rana chensinensis) to test whether the Tsinling Mountains and the nearby Daba Mountains impose major barriers to gene flow. Results Using 13 polymorphic microsatellite DNA loci, 523 individuals from 12 breeding sites with geographical distances ranging from 2.6 to 422.8 kilometers were examined. Substantial genetic diversity was detected at all sites with an average of 25.5 alleles per locus and an expected heterozygosity ranging from 0.504 to 0.855, and two peripheral populations revealed significantly lower genetic diversity than the central populations. In addition, the genetic differentiation among the central populations was statistically significant, with pairwise FST values ranging from 0.0175 to 0.1625 with an average of 0.0878. Furthermore, hierarchical AMOVA analysis attributed most genetic variation to the within-population component, and the between-population variation can largely be explained by isolation-by-distance. None of the putative barriers detected from genetic data coincided with the location of the Tsinling Mountains. Conclusion The Tsinling and Daba Mountains revealed no significant impact on the population genetic structure of R. chensinensis. High population connectivity and extensive juvenile dispersal may account for the significant, but moderate differentiation between populations. Chinese wood frogs are able to use streams as breeding sites at high elevations, which may

  16. Big mountains but small barriers: population genetic structure of the Chinese wood frog (Rana chensinensis) in the Tsinling and Daba Mountain region of northern China.

    PubMed

    Zhan, Aibin; Li, Cheng; Fu, Jinzhong

    2009-04-09

    Amphibians in general are poor dispersers and highly philopatric, and landscape features often have important impacts on their population genetic structure and dispersal patterns. Numerous studies have suggested that genetic differentiation among amphibian populations are particularly pronounced for populations separated by mountain ridges. The Tsinling Mountain range of northern China is a major mountain chain that forms the boundary between the Oriental and Palearctic zoogeographic realms. We studied the population structure of the Chinese wood frog (Rana chensinensis) to test whether the Tsinling Mountains and the nearby Daba Mountains impose major barriers to gene flow. Using 13 polymorphic microsatellite DNA loci, 523 individuals from 12 breeding sites with geographical distances ranging from 2.6 to 422.8 kilometers were examined. Substantial genetic diversity was detected at all sites with an average of 25.5 alleles per locus and an expected heterozygosity ranging from 0.504 to 0.855, and two peripheral populations revealed significantly lower genetic diversity than the central populations. In addition, the genetic differentiation among the central populations was statistically significant, with pairwise FST values ranging from 0.0175 to 0.1625 with an average of 0.0878. Furthermore, hierarchical AMOVA analysis attributed most genetic variation to the within-population component, and the between-population variation can largely be explained by isolation-by-distance. None of the putative barriers detected from genetic data coincided with the location of the Tsinling Mountains. The Tsinling and Daba Mountains revealed no significant impact on the population genetic structure of R. chensinensis. High population connectivity and extensive juvenile dispersal may account for the significant, but moderate differentiation between populations. Chinese wood frogs are able to use streams as breeding sites at high elevations, which may significantly contribute to the

  17. 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

  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. 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...

  20. 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

  1. Population and life-stage-specific effects of two herbicide formulations on the aquatic development of European common frogs (Rana temporaria).

    PubMed

    Wagner, Norman; Veith, Michael; Lötters, Stefan; Viertel, Bruno

    2017-01-01

    Environmental contamination is suggested to contribute to amphibian population declines. However, the effects of a contaminant on a particular amphibian species can differ among populations. The authors investigated the toxic effects of 2 herbicide formulations on different populations and on representative developmental stages of the European common frog (Rana temporaria). Larvae from forest populations were more sensitive to a commonly used glyphosate-based herbicide compared with individuals from agrarian land. Median lethal concentrations correlated with measured glyphosate levels in the breeding ponds, which may be a sign of evolved tolerances. The reverse result was observed for a less commonly used cycloxydim-based herbicide. Effects of the glyphosate-based herbicide were stronger for earlier larval stages compared with later larval stages. Hence, applications in early spring (when early larvae are present in breeding ponds) pose greater risk concerning acute toxic effects on R. temporaria. With regard to late larval stages, short exposure (96 h) of prometamorphic larvae prolonged time to metamorphosis, but only at the highest test concentration that did not significantly induce mortality. This could be due to impairment of the thyroid axis. Notably, nearly all test concentrations of the 2 herbicides provoked growth retardation. Further research on how evolved or induced tolerances are acquired, actual contamination levels of amphibian habitats, and potential endocrine effects of glyphosate-based herbicides is necessary. Environ Toxicol Chem 2017;36:190-200. © 2016 SETAC. © 2016 SETAC.

  2. 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

  3. Comparative analysis of the circadian rhythm genes period and timeless in Culex pipiens Linnaeus, 1758 (Diptera, Culicidae)

    PubMed Central

    Shaikevich, Elena V.; Karan, Ludmila S.; Fyodorova, Marina V.

    2016-01-01

    Abstract Nucleotide sequences of the circadian rhythm genes, period and timeless, were studied for the first time in mosquitoes Culex pipiens Linnaeus, 1758. In this work we evaluated variations of the studied genome fragments for the two forms of Culex pipiens (forma “pipiens” – mosquitoes common for aboveground habitats, forma “molestus” – underground mosquitoes). We compared Culex pipiens from Russia with transatlantic Culex pipiens and subtropical Culex quinquefasciatus Say, 1823. Our results show that intraspecies variability is higher for the gene period than for the gene timeless. The revealed substitutions in nucleotide sequences and especially in amino acid sequences grouped the individuals of the two forms into distinct clusters with high significance. The detected fixed amino acid substitutions may appear essential for functioning of the circadian rhythm proteins in Culex pipiens, and may be correlated with adaptations of the taxa within the group Culex pipiens. Our results suggest that natural selection favors fixed mutations and the decrease in diversity of the genes period and timeless in mosquitoes of the Culex pipiens f. “molestus” compared with the Culex pipiens f. “pipiens”, is probably correlated with adaptive features of Culex pipiens f. “molestus”. The studied genome regions may be considered as promising molecular-genetic markers for identification, population and phylogenetic analysis of similar species and forms of the Culex pipiens complex. PMID:28123673

  4. 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.

  5. 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.

  6. Effects of nutrition and density in Culex pipiens.

    PubMed

    Alto, B W; Muturi, E J; Lampman, R L

    2012-12-01

    Mosquito larvae face numerous biotic and abiotic challenges that affect their development and survivorship, as well as adult fitness. We conducted two experiments under semi-natural conditions to evaluate the effects of intraspecific competition, nutrient limitation and sub-lethal doses of malathion on individual life history traits in adult Culex pipiens (Diptera: Culicidae). In the first experiment, larvae of Cx. pipiens were reared at different intraspecific densities and exposed to sub-lethal doses of malathion. In the second experiment, different intraspecific densities of Cx. pipiens larvae were reared under conditions of low or high larval nutrients, and subsequent adults were fed on either water or 10% sucrose solution. Malathion treatment had relatively minor effects compared with density, which had significant negative effects on development rate, survivorship to adulthood, body size (wing length) and longevity. As larval density increased, a sex ratio distortion in survivorship to adulthood emerged, in which a bias towards males was apparent. Nutrient-rich larval environments alleviated, in part, the effects of increasing density and extended the lifespan of mosquitoes fed on water and 10% sucrose. Density-dependent alterations in adult longevity attributable to the larval environment are complex and show contrasting results depending on interactions with other environmental factors. This study suggests that larval resource availability and competition influence Cx. pipiens population growth correlates and have lasting effects on traits that relate to a mosquito's ability to vector pathogens.

  7. Evaluation of Culex pipiens Populations in a Residential Area with a High Density of Catch Basins in a Suburb of Chicago, Illinois.

    PubMed

    Harbison, Justin E; Henry, Marlon; Xamplas, Christopher; Dugas, Lara R

    2014-09-01

    The North Shore Mosquito Abatement District applies extended release larvicides including methoprene-based Altosid® XR Extended Residual Briquets to approximately 40,000 catch basins in the southern half of the District's operational area at the beginning of each season. Treatments begin in May and typically again 9 to 10 wk later when larvicide efficacy appears to wane. In 2013 spinosad-based Natular™ XRT tablets were applied to basins, and a subset were monitored for larvae and pupae weekly with a standard dipper. When setting the threshold for retreatment as 12 juveniles per dip sample it was observed that basins required a second application 9 wk after the initial application, a time period similar to Altosid despite utilizing a different active ingredient. Average counts of weekly larval samples appeared to be positively associated with average numbers of Culex pipiens collected the following week in a gravid trap located among catch basins, highlighting the importance of basins as sources of these mosquitoes.

  8. Structure, Spatial and Temporal Distribution of the Culex pipiens Complex in Shanghai, China

    PubMed Central

    Gao, Qiang; Xiong, Chenglong; Su, Fei; Cao, Hui; Zhou, Jianjun; Jiang, Qingwu

    2016-01-01

    Background: Culex pipiens molestus was first reported in Shanghai in 2010. The population structures and seasonal distributions of Culex pipiens subspecies C. p. molestus, Culex pipiens pallens, and Culex pipiens quinquefasciatus are not well known. Methods: From late February to November 2013, we conducted daily field surveillance of mosquitoes at eight sites at two green lands and three residential areas in downtown Shanghai. Morphological comparison and DV/D ratios (DV/D is an indicator of mosquito taxonomy) were used to identify adult mosquitoes. Results: The distribution curves of the Culex pipiens complex members indicated seasonal fluctuations. The temperature range of 20–25 °C was the most suitable for adult activity. Micro-environmental factors may differentiate the complex population structures. Hybridization between C. p. pallens and C. p. quinquefasciatus was common and neither “DV/D = 0.40” nor “DV/D = 0.50” can distinguish these subspecies and their hybrids. Conclusion: the population structure of the Culex pipiens complex is complex and characterized by significant hybridization. Measures other than DV/D ratios are needed for the discrimination of subspecies. The C. p. molestus invasion might result in the transmission of novel vector-borne diseases in Shanghai. PMID:27869687

  9. 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

  10. Latitudinal Diversity of Culex pipiens Biotypes and Hybrids in Farm, Peri-Urban, and Wetland Habitats in Europe

    PubMed Central

    Melsen, Diede; Favia, Guido; Wennergren, Uno; Koenraadt, Constantianus J. M.

    2016-01-01

    Despite the presence of Culex (Cx.) pipiens mosquitoes and circulation of West Nile virus (WNV), WNV outbreaks have so far not occurred in northern Europe. The species Cx. pipiens consists of two morphologically identical biotypes, pipiens and molestus, which can form hybrids. Until now, population dynamic studies of Cx. pipiens have not differentiated between biotypes and hybrids at the European scale, nor have they used comparative surveillance approaches. We therefore aimed to elucidate the relative abundance of Cx. pipiens biotypes and hybrids in three habitat types at different latitudes across Europe, using two different surveillance traps. BG-Sentinel and Mosquito-Magnet Liberty Plus traps were placed in three habitat types (farms, peri-urban, wetlands), in three European countries (Sweden, The Netherlands, Italy). Collected Cx. pipiens mosquitoes were identified to biotype with real-time PCR. Both trap types collected equal ratios of the biotypes and their hybrids. From northern to southern latitudes there was a significant decrease of pipiens and an increase of molestus. Habitat types influenced the relative ratios of biotypes and hybrids, but results were not consistent across latitudes. Our results emphasize the need to differentiate Cx. pipiens to the biotype level, especially for proper future WNV risk assessments for Europe. PMID:27870890

  11. The Culex pipiens complex in continental Portugal: distribution and genetic structure.

    PubMed

    Gomes, Bruno; Parreira, Ricardo; Sousa, Carla A; Novo, Maria T; Almeida, António P G; Donnelly, Martin J; Pinto, João

    2012-12-01

    Portugal is a southern European country that displays favorable ecological conditions for the establishment of West Nile virus (WNV) transmission cycles. Competent mosquito vector species are present throughout the country. Among the species with reported cases of WNV isolation in Portugal, Culex pipiens is the most ubiquitous and abundant mosquito. This species exhibits two biological forms with differences in host preferences. The molestus form has a greater tendency to feed upon humans and other mammals whereas the pipiens form prefers avian hosts. In northern latitudes, both forms are physically separated, with molestus occupying underground habitats and pipiens being found aboveground. However, the warmer climatic conditions of southern regions such as Portugal may favor the sympatric occurrence of both forms hence promoting interform hybridization. Genetic introgression between molestus and pipiens forms may result in a higher propensity for admixed populations to serve as bridge-vectors of WNV between humans and birds. Here we revise our present knowledge on the distribution, role in WNV transmission and genetic structure of the Cx. pipiens complex in continental Portugal. We focus on recent findings of sympatric molestus and pipiens populations that display considerable levels of hybridization and discuss the epidemiological repercussions of this occurrence.

  12. 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

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

    PubMed

    Diaz-Badillo, Alvaro; Bolling, Bethany G; Perez-Ramirez, Gerardo; Moore, Chester G; Martinez-Munoz, Jorge P; Padilla-Viveros, America A; Camacho-Nuez, Minerva; Diaz-Perez, Alfonso; Beaty, Barry J; Munoz, Maria de Lourdes

    2011-05-09

    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. 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. 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 collected and identified. The

  14. Summary of 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

    A symposium on the mosquitoes in the Culex pipiens Complex (Pipiens Complex), including Cx. pipiens, Cx. quinquefasciatus, and Cx. molestus, invited 17 of the world's experts on this species Complex to explore the current state of knowledge on the systematic status, speciation, ecology, biology, vector competence, population genetics, endosymbionts, and role in disease transmission of these species in various parts of the world. A Foreword, Introduction, 14 full papers, 3 abstracts and this Summary are published in this special issue of the Journal of the American Mosquito Control Association. A summary of findings, overall conclusions, and research needs will be discussed in relation to mosquito control and interrupting disease transmission caused by these species.

  15. Insecticide resistance to organophosphates in Culex pipiens complex from Lebanon

    PubMed Central

    2012-01-01

    Background Analysis of Culex pipiens mosquitoes collected from a single site in Lebanon in 2005, revealed an alarming frequency of ace-1 alleles conferring resistance to organophosphate insecticides. Following this, in 2006 the majority of municipalities switched to pyrethroids after a long history of organophosphate usage in the country; however, since then no studies have assessed the impact of changing insecticide class on the frequency of resistant ace-1 alleles in C. pipiens. Methods C. pipiens mosquitoes were captured indoors from 25 villages across the country and subjected to established methods for the analysis of gene amplification at the Ester locus and target site mutations in ace-1 gene that confer resistance to organophosphates. Results We conducted the first large-scale screen for resistance to organosphosphates in C. pipiens mosquitoes collected from Lebanon. The frequency of carboxylesterase (Ester) and ace-1 alleles conferring resistance to organophosphates were assessed among C. pipiens mosquitoes collected from 25 different villages across the country between December 2008 and December 2009. Established enzymatic assay and PCR-based molecular tests, both diagnostic of the major target site mutations in ace-1 revealed the absence of the F290V mutation among sampled mosquitoes and significant reduction in the frequency of G119S mutation compared to that previously reported for mosquitoes collected from Beirut in 2005. We also identified a new duplicated ace-1 allele, named ace-1D13, exhibiting a resistant phenotype by associating a susceptible and a resistant copy of ace-1 in a mosquito line sampled from Beirut in 2005. Fisher’s exact test on ace-1 frequencies in the new sample sites, showed that some populations exhibited a significant excess of heterozygotes, suggesting that the duplicated allele is still present. Starch gel electrophoresis indicated that resistance at the Ester locus was mainly attributed to the Ester2 allele, which exhibits

  16. 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...

  17. 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...

  18. First Nationwide Surveillance of Culex pipiens Complex and Culex torrentium Mosquitoes Demonstrated the Presence of Culex pipiens Biotype pipiens/molestus Hybrids in Germany

    PubMed Central

    Börstler, Jessica; Melaun, Christian; Jöst, Hanna; von Thien, Heidrun; Badusche, Marlis; Becker, Norbert; Schmidt-Chanasit, Jonas; Krüger, Andreas; Tannich, Egbert; Becker, Stefanie

    2013-01-01

    Mosquitoes and other arthropods may transmit medically important pathogens, in particular viruses such as West Nile virus. The presence of suitable hosts and competent vectors for those zoonotic viruses is essential for an enzootic transmission, which is a prerequisite for epidemics. To establish reliable risk projections, it is an urgent need for an exact identification of mosquito species, which is especially challenging in the case of sibling species, such as Culex. pipiens pipiens biotypes pipiens and molestus. To facilitate detection of different Culex pipiens forms and their hybrids we established a multiplex real-time PCR. Culex pipiens samples were obtained by egg raft collection and rearing until imago stage or adult sampling using CO2 baited traps and gravid traps. In total, we tested more than 16,500 samples collected all over Germany in the years 2011 and 2012. The predominant species in Germany are Culex pipiens pipiens biotype pipiens and Culex. torrentium, but we also detected Culex pipiens pipiens biotype molestus and hybrids of the two pipiens biotypes at sites where both species occur sympatrically. This report of a potentially important bridge vector for West Nile virus might have major impact in the risk projections for West Nile virus in Germany. PMID:24039724

  19. European Aedes albopictus and Culex pipiens Are Competent Vectors for Japanese Encephalitis Virus.

    PubMed

    de Wispelaere, Mélissanne; Desprès, Philippe; Choumet, Valérie

    2017-01-01

    Japanese encephalitis virus (JEV) is the causative agent of Japanese encephalitis, the leading cause of viral encephalitis in Asia. JEV transmission cycle involves mosquitoes and vertebrate hosts. The detection of JEV RNA in a pool of Culex pipiens caught in 2010 in Italy raised the concern of a putative emergence of the virus in Europe. We aimed to study the vector competence of European mosquito populations, such as Cx. pipiens and Aedes albopictus for JEV genotypes 3 and 5. After oral feeding on an infectious blood meal, mosquitoes were dissected at various times post-virus exposure. We found that the peak for JEV infection and transmission was between 11 and 13 days post-virus exposure. We observed a faster dissemination of both JEV genotypes in Ae. albopictus mosquitoes, when compared with Cx. pipiens mosquitoes. We also dissected salivary glands and collected saliva from infected mosquitoes and showed that Ae. albopictus mosquitoes transmitted JEV earlier than Cx. pipiens. The virus collected from Ae. albopictus and Cx. pipiens saliva was competent at causing pathogenesis in a mouse model for JEV infection. Using this model, we found that mosquito saliva or salivary glands did not enhance the severity of the disease. In this study, we demonstrated that European populations of Ae. albopictus and Cx. pipiens were efficient vectors for JEV transmission. Susceptible vertebrate species that develop high viremia are an obligatory part of the JEV transmission cycle. This study highlights the need to investigate the susceptibility of potential JEV reservoir hosts in Europe, notably amongst swine populations and local water birds.

  20. European Aedes albopictus and Culex pipiens Are Competent Vectors for Japanese Encephalitis Virus

    PubMed Central

    Desprès, Philippe

    2017-01-01

    Background Japanese encephalitis virus (JEV) is the causative agent of Japanese encephalitis, the leading cause of viral encephalitis in Asia. JEV transmission cycle involves mosquitoes and vertebrate hosts. The detection of JEV RNA in a pool of Culex pipiens caught in 2010 in Italy raised the concern of a putative emergence of the virus in Europe. We aimed to study the vector competence of European mosquito populations, such as Cx. pipiens and Aedes albopictus for JEV genotypes 3 and 5. Findings After oral feeding on an infectious blood meal, mosquitoes were dissected at various times post-virus exposure. We found that the peak for JEV infection and transmission was between 11 and 13 days post-virus exposure. We observed a faster dissemination of both JEV genotypes in Ae. albopictus mosquitoes, when compared with Cx. pipiens mosquitoes. We also dissected salivary glands and collected saliva from infected mosquitoes and showed that Ae. albopictus mosquitoes transmitted JEV earlier than Cx. pipiens. The virus collected from Ae. albopictus and Cx. pipiens saliva was competent at causing pathogenesis in a mouse model for JEV infection. Using this model, we found that mosquito saliva or salivary glands did not enhance the severity of the disease. Conclusions In this study, we demonstrated that European populations of Ae. albopictus and Cx. pipiens were efficient vectors for JEV transmission. Susceptible vertebrate species that develop high viremia are an obligatory part of the JEV transmission cycle. This study highlights the need to investigate the susceptibility of potential JEV reservoir hosts in Europe, notably amongst swine populations and local water birds. PMID:28085881

  1. Falcaustra lowei n. sp. and other helminths from the Tarahumara frog, Rana tarahumarae (Anura: Ranidae), from Sonora, Mexico.

    PubMed

    Bursey, C R; Goldberg, S R

    2001-04-01

    Seventy-four specimens of Falcaustra lowei n. sp. were recovered from the intestines of 9 of 42 (21%) Tarahumara frogs. Rana tarahumarae, from Sonora, Mexico. F. lowei is the 14th Nearctic species to be described and belongs to that group of species possessing a pseudosucker, namely F. catesbeianae, F. chabaudi, F. chelydrae, F. mexicana, and F. wardi. The new species can be readily differentiated from these by the arrangement of caudal papillae and length of spicules. Priority description of F. affinis is established and F. concinnae is removed from synonymy with F. affinis. In addition to F. lowei, 3 species of Digenea, Glypthelmins quieta, Haematoloechus breviplexus, Langeronia macrocirra; 1 species of Eucestoda, Ophiotaenia magna; 7 species of Nematoda, F. inglisi, Foleyellides striatus, Oswaldocruzia pipiens, Rhabdias ranae, Subulascaris falcaustriformis, Physaloptera sp. (larvae): and 1 species of Acanthocephala, an unidentified oligacanthorhynchid cystacanth, were found.

  2. Midgut Barrier Imparts Selective Resistance to Filarial Worm Infection in Culex pipiens pipiens

    PubMed Central

    Michalski, Michelle L.; Erickson, Sara M.; Bartholomay, Lyric C.; Christensen, Bruce M.

    2010-01-01

    Mosquitoes in the Culex pipiens complex thrive in temperate and tropical regions worldwide, and serve as efficient vectors of Bancroftian lymphatic filariasis (LF) caused by Wuchereria bancrofti in Asia, Africa, the West Indies, South America, and Micronesia. However, members of this mosquito complex do not act as natural vectors for Brugian LF caused by Brugia malayi, or for the cat parasite B. pahangi, despite their presence in South Asia where these parasites are endemic. Previous work with the Iowa strain of Culex pipiens pipiens demonstrates that it is equally susceptible to W. bancrofti as is the natural Cx. p. pipiens vector in the Nile Delta, however it is refractory to infection with Brugia spp. Here we report that the infectivity barrier for Brugia spp. in Cx. p. pipiens is the mosquito midgut, which inflicts internal and lethal damage to ingested microfilariae. Following per os Brugia exposures, the prevalence of infection is significantly lower in Cx. p. pipiens compared to susceptible mosquito controls, and differs between parasite species with <50% and <5% of Cx. p. pipiens becoming infected with B. pahangi and B. malayi, respectively. When Brugia spp. mf were inoculated intrathoracically to bypass the midgut, larvae developed equally well as in controls, indicating that, beyond the midgut, Cx. p. pipiens is physiologically compatible with Brugia spp. Mf isolated from Cx. p. pipiens midguts exhibited compromised motility, and unlike mf derived from blood or isolated from the midguts of Ae. aegypti, failed to develop when inoculated intrathoracically into susceptible mosquitoes. Together these data strongly support the role of the midgut as the primary infection barrier for Brugia spp. in Cx. p. pipiens. Examination of parasites recovered from the Cx. p. pipiens midgut by vital staining, and those exsheathed with papain, suggest that the damage inflicted by the midgut is subcuticular and disrupts internal tissues. Microscopic studies of these worms

  3. 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...

  4. 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...

  5. Variation in adult longevity of Culex pipiens f. pipiens, vector of the West Nile Virus.

    PubMed

    Andreadis, S S; Dimotsiou, O C; Savopoulou-Soultani, M

    2014-11-01

    The common house mosquito, Culex pipiens (Diptera: Culicidae), which is considered the primary bridge vector of West Nile Virus (WNV) to humans, is a wide spread insect pest with medical importance and consists of two distinct bioforms, Cx. pipiens f. pipiens and Cx. pipiens f. molestus. Here, we consider the adult lifespan of male and female Cx. pipiens f. pipiens under controlled conditions at five constant temperature regimes (15, 20, 25, 27.5, and 30 °C). Our results show that adult longevity was affected by temperature, as it significantly decreased with increase in temperature. At the highest tested temperature, mean adult longevity did not exceed 12 days for both sexes and thus makes impossible the risk of WNV transmission. On the other hand at the lowest temperature, longevity was extremely high with some individuals surviving up to 129 and 132 days, males and females, respectively, and thus enable them to function as potential vectors of WNV for a prolonged period of time. As far as sex is concerned, adult females displayed a 1.2-1.4-fold longer longevity compared to the male ones. However, this difference was significant only at the lowest and highest tested temperature regime. This information is useful in determining the critical temperatures which may affect the distribution of Cx. pipiens and consequently the risk of WNV transmission. Moreover, the effect of environmental temperature should be considered when evaluating the abundance of these species.

  6. Survey and assessment of amphibian populations in Rocky Mountain National Park

    USGS Publications Warehouse

    Corn, Paul Stephen; Jennings, Michael L.; Muths, Erin L.

    1997-01-01

    We conducted surveys in Rocky Mountain National Park, Colorado for amphibians in 1987-1994. Four species, Ambystoma tigrinum, Bufo boreas, Pseudacris maculata, and Rana sylvatica, were recorded. Pseudacris maculata was the most widely distributed and abundant species in the Park. Two populations of E maculata were estimated to contain 161 and 136 breeding males in 1988. There was no evidence of a decline of A. tigrinum or R. sylvatica, but these species were found at relatively few locations. We did not detect Rana pipiens, which had been known previously from 3 locations in the Park. We found 7 breeding populations of B. boreas, which has declined recently elsewhere in the southern Rocky Mountains, but all but 2 of these populations were small and may not reproduce annually. At least one of these small populations is thought to have been extirpated. Estimated numbers of males in the 2 large populations, which are 6.4 km apart in the same drainage, were stable or increasing slightly from 1992 to 1995, averaging 189 and 239 individuals. Current and known locations of amphibians did not differ in elevation, size, lake type, presence of shallow water or emergent vegetation on the north shore, or presence of trout. Water chemistry at amphibian breeding sites was variable, but pH decreased significantly with increasing elevation. Causes of declines of B. boreas and R. pipiens are not known. Populations of B. boreas in the North Fork of the Big Thompson River are critically important to the conservation of this species in the Rocky Mountains.

  7. 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

  8. High Insecticides Resistance in Culex pipiens (Diptera: Culicidae) from Tehran, Capital of Iran

    PubMed Central

    Salim-Abadi, Yaser; Oshaghi, Mohammad Ali; Enayati, Ahmad Ali; Abai, Mohammad Reza; Vatandoost, Hassan; Eshraghian, Mohammad Reza; Mirhendi, Hossein; Hanafi-Bojd, Ahmad Ali; Gorouhi, Mohammad Amin; Rafi, Fatemeh

    2016-01-01

    Background: During recent years transmission of Dirofilaria immitis (dog heart worm) by Culex pipiens and West Nile virus have been reported from Iran. The present study was preformed for evaluating the susceptibility status of Cx. pipiens collected from capital city of Tehran, Iran. Methods: Four Insecticides including: DDT 4%, Lambdacyhalothrin 0.05%, Deltamethrin 0.05% and Cyfluthrin 0.15 % according to WHO standard methods were used for evaluating the susceptibility status of Cx. pipiens from Tehran moreover For comparison susceptibility status a Laboratory strain also was used. Bioassay data were analyzed using Probit program. The lethal time for 50% and 90% mortality (LT50 and LT90) values were calculated from regression line. Results: The susceptibility status of lab strain of Cx. pipiens revealed that it is susceptible to Lambdacyhalothrin, Deltamethrin, Cyfluthrin and resistant to DDT. Moreover cyfluthrin with LT50=36 seconds and DDT with LT50=3005 seconds had the least and most LT50s. Field population was resistance to all tested insecticides and DDT yielded no mortality. Conclusion: Highly resistance level against all WHO recommended imagicides were detected in field populations. We suggest more biochemical and molecular investigations to detect resistance mechanisms in the field population for further decision of vector control. PMID:28032100

  9. Experimental host preference of diapause and non-diapause induced Culex pipiens pipiens (Diptera: Culicidae).

    PubMed

    Faraji, Ary; Gaugler, Randy

    2015-07-24

    Culex pipiens pipiens plays an important role in the transmission of several vector-borne pathogens such as West Nile virus (WNV) in North America. Laboratory and field studies suggest that this species is ornithophilic but because of genetic hybridization with sibling species during the active mosquito season, it may occasionally feed on mammals. Adult female Cx. p. pipiens undergo a facultative diapause and may serve as an overwintering mechanism for WNV. To determine the effect of diapause on the innate host preference of Cx. p. pipiens emerging from winter hibernation, we conducted host-choice experiments using bird and mammal hosts. Mosquitoes were reared under non-diapause induced (NDI), diapause induced (DI), and field collected from overwintering (OW) hibernaculae. They were released into a large mesh enclosure housing two lard can traps, and given a choice between feeding on a dove or a rat. Host seeking Cx. p. pipiens were four times more likely to feed on the dove than the rat, regardless of experimental conditions. Under NDI conditions, Cx. p. pipiens were (p < 0.001) more attracted to the bird (79.9% [75.6-84.1]) than the rat (20.1 [15.9-24.4]). Overwintering mosquitoes and those exposed to DI conditions were also significantly (p < 0.001) more attracted to birds (81.6% [75.9-87.3]) than to rats (18.5 [12.7-24.2]). We provide new information about the innate host preference of Cx. p. pipiens emerging from diapause in temperate habitats where winter survival is crucial for disease transmission cycles. Although we showed that Cx. p. pipiens prefers an avian to a mammalian host, nearly 20% of emerging mosquitoes in the spring could feed on mammals. Changes in host preferences may also contain valuable clues about transmission dynamics and subsequent timely interventions by vector control and public health practitioners.

  10. Flushing effect of rain on container-inhabiting mosquitoes Aedes aegypti and Culex pipiens (Diptera: Culicidae).

    PubMed

    Koenraadt, C J M; Harrington, L C

    2008-01-01

    We investigated the role of heavy rain on container-inhabiting mosquito (Diptera: Culicidae) populations, and how different species may have adapted to such conditions. Rains were created with a rain simulator calibrated to natural rain intensities in the habitats of two important vector species: Aedes aegypti (L.) from northern Thailand and Culex pipiens L. from New York state, USA. Immature stages of Ae. aegypti were able to resist the flushing effect of rain better than Cx. pipiens. This difference was most dramatic during the pupal stage. Fourth instars of Ae. aegypti were not affected by flushing when exposed for longer rain intervals (30 versus 60 min) or at a colder water temperature (24 versus 16 degrees C). In contrast, significantly more Cx. pipiens larvae flushed out with longer rain exposure. Warmer water temperatures also increased the proportion of Cx. pipiens flushed out, but mostly at the longest exposure time. Container position (tilted at a 7 degrees angle or level) did not affect proportions of fourth instars flushed out for both species. More accurate models of vector-borne diseases can be developed by incorporating the described effects of rain on container-breeding mosquito populations. Such models may provide more realistic assessments of disease risk and ensure optimal use of limited financial resources of mosquito control agencies.

  11. 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.

  12. Detection and widespread distribution of sodium channel alleles characteristic of insecticide resistance in Culex pipiens complex mosquitoes in China.

    PubMed

    Wang, Z-M; Li, C-X; Xing, D; Yu, Y-H; Liu, N; Xue, R-D; Dong, Y-D; Zhao, T-Y

    2012-06-01

    Culex pipiens complex mosquitoes are widely distributed throughout China and are known to be important disease vectors. Two pyrethroid resistance associated mutations have been identified in Cx. pipiens complex (Diptera: Culicidae), but there is little information on the diversity and distribution of kdr alleles in pyrethroid resistance in Cx. pipiens complex mosquitoes in China. In the present study, we report on a modified three tube allele-specific (AS)-PCR method for detecting the 1014F and 1014S alleles. The new technique was applied to identify the distribution of the two alleles in natural Cx. pipiens complex populations in China. The results confirmed that the new method is both sensitive and specific. The 1014F allele was found in all 14 of the field populations tested (frequency ranged from 6.8 to 76.2%) and the 1014S allele was found in almost two-thirds (frequency from 2.4 to 28.6%), indicating that the genotypes known to be associated with pyrethroid resistance are widespread in China. The resistance-associated alleles were more common in southern Chinese sampling sites than in northern sites. The coexistence of the two resistant mutations in individual mosquitoes was also observed in five of the field populations. Two alternative mutations within the L1014 codon were identified in Culex pipiens molestus Forskal, 1775, including a non-synonymous mutation resulting in a 1014C substitution. © 2011 The Authors. Medical and Veterinary Entomology © 2011 The Royal Entomological Society.

  13. 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...

  14. Possible mechanisms responsible for the reduced intestinal flora in hibernating leopard frogs (Rana pipiens).

    PubMed Central

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

    1988-01-01

    Mechanisms and factors that normally control the large intestinal flora were investigated to determine whether changes in these parameters could account for the decreased bacterial concentration and facultative nature of the flora found in hibernating frogs. It appeared that low temperatures and limited nutrients were the main factors responsible for the decrease in the bacterial concentration and may also have been responsible for the increase in the proportions of facultative organisms, since no change in the redox potential was seen. The hibernating frogs were extremely sluggish in the removal of India ink particles from the circulatory system by the Kupffer cells of the liver compared with nonhibernating frogs. They were unable to mount an antibody response to bovine serum albumin, but their serum did exhibit killing of Pseudomonas paucimobilis, suggesting opsonization by preformed antibody and complement. The role of these host factors in protecting the hibernating frog against this indigenous flora is discussed. PMID:3263839

  15. Pharmacology, Distribution and Development of Muscarinic Acetylcholine Receptor Subtypes in the Optic Tectum of Rana Pipiens

    PubMed Central

    Butt, C. M.; Pauly, J. R.; Wilkins, L. H.; Dwoskin, L. P.; Debski, E. A.

    2008-01-01

    Visually evoked behaviors mediated by the frog optic tectum require cholinergic activity, but the receptor subtypes through which acetylcholine acts are not yet identified. Using quantitative autoradiography and scintillation spectrometry, we examined the binding of [3H]pirenzepine and [3H]AF-DX 384 in the laminated optic tectum of the frog. In mammalian systems, these substances bind excitatory (m1 and m3 subtypes) and inhibitory (m2 and m4 subtypes) muscarinic acetylcholine receptors, respectively. Pharmacological analyses, including the use of specific muscarinic toxins, confirmed the subtype selectivity of the radioligands in the frog brain. Binding sites for [3H]pirenzepine were distinct from those for [3H]AF-DX 384. In the adult tectum, [3H]pirenzepine demonstrated specific binding in tectal layers 5–9. [3H]Pirenzepine binding was also present in tadpoles as young as stage V, but all sampled stages of tadpole tectum had significantly less binding when compared to adults. Lesioning of the optic nerve had no effect on [3H]pirenzepine binding. Specific [3H]AF-DX 384 binding was found in all layers of the adult tectum. All sampled tadpole stages exhibited binding sites for [3H]AF-DX 384, but the densities of these sites were also significantly higher in adults than they were in developing stages. Short-term lesions of the optic nerve reduced [3H]AF-DX 384 binding in all tectal layers of the deafferented lobe when compared to the afferented one. Long-term lesions decreased [3H]AF-DX 384 sites in both lobes. These results indicate that multiple muscarinic acetylcholine receptor binding sites reside in the frog optic tectum at all stages of development, and their pharmacology resembles that of mammalian m1/m3, m2 and m4 subtypes. Our data indicate that few, if any, of these receptors are likely to be located on retinal ganglion cell terminals. Furthermore, the expression of inhibitory muscarinic subtypes seems to be regulated by different mechanisms than that for excitatory subtypes. PMID:11311540

  16. Behavioral effects of morphine, levorphanol, dextrorphan and naloxone in the frog Rana pipiens.

    PubMed

    Pezalla, P D; Stevens, C W

    1984-08-01

    Systemic morphine induces explosive motor behavior and generalized muscular rigidity in frogs. Naloxone does not reverse either of these effects of morphine but at high doses causes muscular flaccidity and unresponsiveness to stimulation. Intraspinal morphine induces rigidity, but not explosive motor behavior, and this action is blocked by naloxone. Behavioral effects are seen rarely after intraspinal levorphanol (rigidity) and never after intraspinal dextrorphan or naloxone. In contrast to systemic morphine and naloxone, systemic levorphanol and dextrorphan are lethal to frogs at high doses.

  17. Effects of external calcium on potassium contractures in tonic muscle fibers of the frog (Rana pipiens).

    PubMed Central

    Huerta, M; Muñiz, J; Stefani, E

    1986-01-01

    K+ contractures of tonic bundles from cruralis muscle of the frog were studied with different K+ concentrations (10-120 mM). K+ contractures had an initial transient phase followed by a sustained tension. The amplitude of the sustained tension diminished with high K+ concentration (80-120 mM). However, in all cases, tension was maintained for several minutes. External Ca2+ reduction practically abolished the sustained phase of the K+ contractures. The initial phase was also reduced and tension spontaneously relaxed. The curve relating the peak tension with log [K+]o, showed that the threshold was not affected but the peak tension was reduced to about 70% in low-Ca2+ saline (0 Ca2+ + 3 mM-Mg2+) and 50% in Ca2+-free saline (1 mM-EGTA + 3 mM-Mg2+). The dependence of the sustained tension on external Ca2+ was further confirmed by Ca2+ withdrawal and re-establishment and/or by Ni2+ substitution for Ca2+ before or during K+ contractures. These results indicate that external Ca2+ had to be continuously present to maintain the tension during K+ contractures and that Ni2+ was not able to restore the normal temporal course of K+ contracture. The sustained phase was diminished by blocking agents of Ca2+ channels, such as nifedipine (1 microM) and diltiazem (1-10 microM). The present results can be explained by a direct control of the Ca2+ currents on K+ contracture or by specific interactions between external Ca2+ and Ca2+-binding sites in the membrane. PMID:3491902

  18. Helminths of two native frog species (Rana chiricahuensis, Rana yavapaiensis) and one introduced frog species (Rana catesbeiana) (Ranidae) from Arizona.

    PubMed

    Goldberg, S R; Bursey, C R; Cheam, H

    1998-02-01

    The gastrointestinal tracts, lungs, urinary bladders, and body cavities of Rana catesbeiana (n = 25), Rana chiricahuensis (n = 25), and Rana yavapaiensis (N = 37) from Arizona were examined for helminths. Helminths representing 9 species of trematodes: Cephalogonimus brevicirrus, Glypthelmins quieta, Gorgoderina attenuata, Haematoloechus complexus, Haematoloechus langiplexus, Megalodiscus temperatus, Alaria sp., Clinostomum sp., and an unidentified strigeid; and 4 species of nematodes: Falcaustra catesbeianae, Rhabdias ranae, Physaloptera sp., and an unidentified ascarid were found. The helminth fauna of introduced R. catesbeiana differed markedly from that of native ranids. Helminths of R. chiricahuensis and R. yavapaiensis represent new host records. Arizona is a new locality record for C. brevicirrus, G. attenuata, H. complexus, H. longiplexus, M. temperatus, and R. ranae.

  19. 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.

  20. Expression of frequenin at the frog (Rana) neuromuscular junction, muscle spindle and nerve.

    PubMed

    Werle, M J; Roder, J; Jeromin, A

    2000-04-21

    Frequenin is a calcium binding protein previously implicated in the regulation of neurotransmission in Drosophila and Xenopus. We have used the frog (Rana pipiens) to study the localization and regulation of expression of frequenin-like molecules in the vertebrate peripheral nervous system. Affinity purified antibodies to frequenin recognize molecules in the neuromuscular junction, axons in the peripheral nerve, and neuronal processes in muscle spindles. Western blots of endplate regions, peripheral nerve, and brain, resulted in the labelling of a single 24 kDa band, which is the expected size for frequenin. These results suggest that frequenin expression is high in the frog peripheral nervous system, and may reflect a function for frequenin in synaptic transmission in vertebrates.

  1. 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.

  2. 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. © 2016 Wiley Periodicals, Inc.

  3. 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

  4. Complete mitochondrial genomes of two brown frogs, Rana dybowskii and Rana cf. chensinensis (Anura: Ranidae).

    PubMed

    Li, Jiao; Lei, Guangchun; Fu, Cuizhang

    2016-01-01

    We first determined complete mitochondrial genomes of Rana dybowskii and Rana cf. chensinensis (Anura: Ranidae). The mitogenomic lengths of R. dybowskii and R. cf. chensinensis were 18,864 and 18,808 bp, respectively. The two mitogenomes have similar gene compositions including 13 protein-coding genes, 22 tRNA genes, 2 rRNA genes and a control region. Rana dybowskii and R. cf. chensinensis mitogenomes displayed same gene order arrangements and similar base compositions with an A + T bias. Mitogenomic data of the two species contributed to provide molecular marker for their conservative genetics and clarified their phylogenetic position under mitogenome-based phylogeny of the genus Rana.

  5. Cane-sugar feeding in Culex pipiens fatigans.

    PubMed

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

    1967-01-01

    The relatively poor results that have been obtained in controlling or eradicating Culex pipiens fatigans, the urban vector of Wuchereria bancrofti, have made it necessary to obtain as much information as possible about its biology. In this paper the unexpected finding of the influence of cane sugar in delaying oviposition in the gravid female is reported. This is an important finding for those who are investigating the biology of this mosquito in the laboratory and who make use of the age-old practice of keeping adult mosquitos alive by feeding them cane sugar. It is also reported that under certain conditions extensive cane-sugar feeding takes place in nature and it seems possible that this habit may have unsuspected repercussions on behaviour and physiology beyond the confines of the laboratory. One possible development would be the isolation of the attractant present in unrefined sugars and its use in traps in order to assess mosquito populations in the field.

  6. 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...

  7. 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...

  8. Hypervariable prophage WO sequences describe an unexpected high number of Wolbachia variants in the mosquito Culex pipiens.

    PubMed

    Duron, Olivier; Fort, Philippe; Weill, Mylène

    2006-02-22

    Wolbachia are maternally inherited endosymbiotic bacteria that infect many arthropod species and may induce cytoplasmic incompatibility (CI) resulting in abortive embryonic development. Among all the described host species, mosquitoes of the Culex pipiens complex display the highest variability of CI crossing types. Paradoxically, searches for polymorphism in Wolbachia infecting strains and field populations hitherto failed or produced very few markers. Here, we show that an abundant source of the long-sought polymorphism lies in WO prophage sequences present in multiple copies dispersed in the genome of Wolbachia infecting C. pipiens (wPip). We identified up to 66 different Wolbachia variants in C. pipiens strains and field populations and no occurrence of superinfection was observed. At least 49 different Wolbachia occurred in Southern Europe C. pipiens populations, and up to 10 different Wolbachia were even detected in a single population. This is in sharp contrast with North African and Cretan samples, which exhibited only six variants. The WO polymorphism appeared stable over time, and was exclusively transferred maternally. Interestingly, we found that the CI pattern previously described correlates with the variability of Gp15, a prophage protein similar to a bacterial virulence protein. WO prophage sequences thus represent variable markers that now open routes for approaching the molecular basis of CI, the host effects, the structure and dynamics of Wolbachia populations.

  9. Hypervariable prophage WO sequences describe an unexpected high number of Wolbachia variants in the mosquito Culex pipiens

    PubMed Central

    Duron, Olivier; Fort, Philippe; Weill, Mylène

    2005-01-01

    Wolbachia are maternally inherited endosymbiotic bacteria that infect many arthropod species and may induce cytoplasmic incompatibility (CI) resulting in abortive embryonic development. Among all the described host species, mosquitoes of the Culex pipiens complex display the highest variability of CI crossing types. Paradoxically, searches for polymorphism in Wolbachia infecting strains and field populations hitherto failed or produced very few markers. Here, we show that an abundant source of the long-sought polymorphism lies in WO prophage sequences present in multiple copies dispersed in the genome of Wolbachia infecting C. pipiens (wPip). We identified up to 66 different Wolbachia variants in C. pipiens strains and field populations and no occurrence of superinfection was observed. At least 49 different Wolbachia occurred in Southern Europe C. pipiens populations, and up to 10 different Wolbachia were even detected in a single population. This is in sharp contrast with North African and Cretan samples, which exhibited only six variants. The WO polymorphism appeared stable over time, and was exclusively transferred maternally. Interestingly, we found that the CI pattern previously described correlates with the variability of Gp15, a prophage protein similar to a bacterial virulence protein. WO prophage sequences thus represent variable markers that now open routes for approaching the molecular basis of CI, the host effects, the structure and dynamics of Wolbachia populations. PMID:16615218

  10. Composite linkage map and enhanced genome map for Culex pipiens complex mosquitoes.

    PubMed

    Hickner, Paul V; Mori, Akio; Chadee, Dave D; Severson, David W

    2013-01-01

    We report here the development of 65 novel microsatellite loci and construction of a composite genetic linkage map for Culex pipiens complex mosquitoes. Microsatellites were identified by in silico screening of the Culex quinquefasciatus genome assembly. Cross-species utility of 73 microsatellites for population studies in C. pipiens sensu stricto and C. quinquefasciatus was evaluated by genotyping a subset of samples collected in Indiana, United States, and Point Fortin, Trinidad. Allele frequencies of 67 microsatellites were within Hardy-Weinberg expectations in both population subsets. A composite linkage map was constructed based on restriction fragment length polymorphism and microsatellite polymorphisms in 12 independent F1 intercross mapping populations. The composite map consists of 61 marker loci totaling 183.9 cM distributed across the 3 linkage groups. These loci cover 29.5, 88.8, and 65.6 cM on chromosomes I-III, respectively, and allow for assignment of 10.4% of the genome assembly and 13.5% of the protein coding genes to chromosome position. Our results suggest that these microsatellites will be useful for mapping and population studies of 2 pervasive species in the C. pipiens complex. Moreover, the composite map presented here will serve as a basis for the construction of high-resolution genetic and physical maps, as well as detection of quantitative trait loci to aid in the investigation of complex genetic traits influencing phenotypes of interest.

  11. Identification of individual foothill yellow-legged frogs (Rana boylii) using chin pattern photographs: a non-invasive and effective method for small population studies

    Treesearch

    K.R. Marlow; K.D. Wiseman; Clara Wheeler; J.E.  Drennan; R.E.  Jackman

    2016-01-01

    The ability to identify individual animals is a valuable tool in the study of amphibian population dynamics, movement ecology, social behavior, and habitat use. Numerous methods of marking amphibians have been employed including the use of passive integrated transponder (PIT) tags, radio-transmitters, elastomers, branding, and mutilation techniques such as toe...

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

    PubMed

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

    2015-07-01

    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. © The Authors 2015. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  13. Microsatellite primers for Culex pipiens quinquefasciatus, the vector of avian malaria in Hawaii

    USGS Publications Warehouse

    Fonseca, Dina M.; Atkinson, Carter T.; Fleischer, Robert C.

    1998-01-01

    The southern house mosquito, Culex pipiens quinquefasciatus (Diptera: Culicidae), was introduced accidentally to Hawaii in 1826 (van Riper et al. 1986). There it eventually became the vector of avian malaria, Plasmodium relictum, a disease that severely limits the size and distribution of endemic forest bird populations in Hawaii (Atkinson et al. 1995). Cx.p. quinquefasciatus has a circumtropical distribution and is also the vector for human diseases such as lymphatic filariasis and several encephalitis.

  14. Exposure of leopard frogs to a pesticide mixture affects life history characteristics of the lungworm Rhabdias ranae.

    PubMed

    Gendron, A D; Marcogliese, D J; Barbeau, S; Christin, M-S; Brousseau, P; Ruby, S; Cyr, D; Fournier, M

    2003-05-01

    We tested the hypothesis that exposure of leopard frogs ( Rana pipiens) to agricultural pesticides can affect the infection dynamics of a common parasite of ranid frogs, the lungworm Rhabdias ranae. After a 21-day exposure to sublethal concentrations of a pesticide mixture composed of atrazine, metribuzin, aldicarb, endosulfan, lindane and dieldrin, or to control solutions (water, dimethyl sulfoxide), parasite-free juvenile frogs were challenged with 30 infective larvae of R. ranae. Approximately 75% of the larvae penetrated the skin and survived in both exposed and control animals, suggesting that pesticides did not influence host recognition or penetration components of the transmission process. Rather, we found that the migration of R. ranae was significantly accelerated in hosts exposed to the highest concentrations of pesticides, leading to the establishment of twice as many adult worms in the lungs of frogs 21 days post-infection. Pesticide treatment did not influence the growth of lungworms but our results indicate that they matured and reproduced earlier in pesticide-exposed frogs compared to control animals. Such alterations in life history characteristics that enhance parasite transmission may lead to an increase in virulence. Supporting evidence shows that certain components of the frog immune response were significantly suppressed after exposure to the pesticide mixture. This suggests that the immune system of anurans exerts a control over lungworm migration and maturation and that agricultural contaminants can interfere with these control mechanisms. Our results also contribute to the ongoing debate regarding the role that anthropogenic factors could play in the perplexing disease-related die-offs of amphibians observed in several parts of the world.

  15. High Degree of Single Nucleotide Polymorphisms in California Culex pipiens (Diptera: Culicidae) sensu lato

    PubMed Central

    LEE, YOOSOOK; SEIFERT, STEPHANIE N.; NIEMAN, CATELYN C.; McABEE, RORY D.; GOODELL, PARKER; FRYXELL, REBECCA TROUT; LANZARO, GREGORY C.; CORNEL, ANTHONY J.

    2013-01-01

    Resolution of systematic relationships among members of the Culex pipiens (L.) complex has important implications for public health as well as for studies on the evolution of sibling species. Currently held views contend that in California considerable genetic introgression occurs between Cx. pipiens and Cx. quinquefasciatus Say, and as such, these taxa behave as if they are a single species. Development of high throughput SNP genotyping tools for the analysis of Cx. pipiens complex population structure is therefore desirable. As a first step toward this goal, we sequenced 12 gene fragments from specimens collected in Marin and Fresno counties. On average, we found a higher single nucleotide polymorphism (SNP) density than any other mosquito species reported thus far. Coding regions contained significantly higher GC content (median 54.7%) than noncoding regions (42.4%; Wilcoxon rank sum test, P = 5.29 × 10−5). Differences in SNP allele frequencies observed between mosquitoes from Marin and Fresno counties indicated significant genetic divergence and suggest that SNP markers will be useful for future detailed population genetic studies of this group. The high density of SNPs highlights the difficulty in identifying species within the complex and may be associated with the large degree of phenotypic variation observed in this group of mosquitoes. PMID:22493847

  16. Insecticide-resistance in Culex pipiens fatigans*

    PubMed Central

    Busvine, J. R.

    1967-01-01

    The three main subspecies of the Culex pipiens complex—pipiens, fatigans and molestus—have similar “normal” levels of susceptibility to insecticides. C. p. fatigans larvae resemble those of anopheline mosquitos in sensitivity to dieldrin, but are more resistant to DDT; the adults are unusually resistant to chlorinated insecticides, but do not differ greatly from other species of mosquito in susceptibility to organophosphorus compounds. There is evidence of conversion of DDT to DDE in C. p. fatigans and other mosquitos, but this may well not be the only cause of resistance. Dieldrin-resistance in C. p. fatigans involves cross-resistance to HCH, but at a considerably lower level; the mechanism of resistance is still obscure. Resistance to organophosphorus insecticides has been observed; the cross-resistance to diazinon indicates that the mechanism is not solely one of carboxy-esterase detoxication. Mechanisms of inheritance of resistance have been investigated. The most recent results indicate the monofactorial inheritance of resistance by a single pair of nearly dominant genes on chromosome 2 for DDT-resistance and by a single pair of genes on chromosome 3 with intermediate dominance for HCH dieldrin-resistance. PMID:5300068

  17. Indirect effects of introduced trout on Cascades frogs (Rana cascadae) via shared aquatic prey

    Treesearch

    Maxwell B. Joseph; Jonah Piovia-Scott; Sharon P. Lawler; Karen L. Pope

    2010-01-01

    1. The introduction of trout to montane lakes has negatively affected amphibian populations across the western United States. In northern California’s Klamath–Siskiyou Mountains, introduced trout have diminished the distribution and abundance of a native ranid frog, Rana (=Lithobates)

  18. Ribose-phosphate pyrophosphokinase 1 (PRPS1) associated with deltamethrin resistance in Culex pipiens pallens

    PubMed Central

    Hong, Shanchao; Zhou, Dan; Chen, Chen; Wang, Weijie; Lv, Yuan; Ye, Yvting; Zou, Ping; Yv, Jing; Chang, Xuelian; Yv, Xinyou; Shi, Linna; Ma, Lei; Sun, Yan; Zhang, Donghui

    2013-01-01

    Ribose-phosphate pyrophosphokinase 1 (PRPS1) was identified and isolated as a differentially expressed gene between deltamethrin-susceptible (DS) and deltamethrin-resistant (DR) Culex pipiens pallens and Aedes albopictus C6/36 cell line through microarray and 2D-Gel. An open reading frame of PRPS1 cloned from C. pipiens pallens has 1,011 bp and encodes for a 336 amino acids protein which shares high homology with Culex quinquefasciatus. Real-time polymerase chain reaction was used to determine the transcript expression level of PRPS1 in DS and DR strains. The expression levels of PRPS1 were higher in DR laboratory strains and natural population JXZ-DR, JXZ-LDR. PRPS1 was also detected and expressed at all developmental stages of C. pipiens pallens and increased expression level in DR3 strain than DS strain in the third and fourth instar larvae, female and male stages. In addition, to further investigate the role of PRPS1 in deltamethrin resistance, PRPS1 was transiently expressed in A. albopictus C6/36 cells and detected by western blotting. Cells transfected with PRPS1 had an increased resistance to deltamethrin compared with control cells. These results suggested that the increased expression level of PRPS1 may play roles in the regulation of deltamethrin resistance. PMID:23250545

  19. 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

  20. 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

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

    PubMed

    Zou, Feifei; Chen, Chen; 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.

  2. 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

  3. 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

    2015-07-07

    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.

  4. 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.

  5. Naive and conditioned responses of Culex pipiens pipiens biotype molestus (Diptera: Culicidae) to flower odors.

    PubMed

    Jhumur, Umma Salma; Dötterl, Stefan; Jürgens, Andreas

    2006-11-01

    Flower odors are important signals for chemical communication between plants and flower visitors. Here, we studied the naive responses of Culex pipiens pipiens biotype molestus Forskal 1775 (Diptera: Culicidae) to typical flower odors and assessed the learning capacity of mosquitoes to floral volatiles. The odor compounds used in the bioassay, phenyl acetaldehyde, veratrole, and 2-methoxyphenol, are typically found in the floral odor of Silene otites (L.) Wibel, a plant that is pollinated by nectar-drinking mosquitoes and moths, and/or in other closely related Silene species. Wind tunnel bioassays with a mixture of these compounds revealed that attraction of mosquitoes to odors was positively correlated with time passed since the last feeding. In single component bioassays, mosquitoes showed strong innate responses to phenyl acetaldehyde and only moderate or weak responses to veratrole and 2-methoxyphenol. Furthermore, in comparison with naive mosquitoes, conditioned mosquitoes were significantly more attracted to the mixture and single volatiles. These results indicate that naive mosquitoes are effectively attracted by appropriate floral scent compounds and that learning can increase the attractiveness of these compounds.

  6. Transmission Incompetence of Culex quinquefasciatus and Culex pipiens pipiens from North America for Zika Virus

    PubMed Central

    Kenney, Joan L.; Romo, Hannah; Duggal, Nisha K.; Tzeng, Wen-Pin; Burkhalter, Kristen L.; Brault, Aaron C.; Savage, Harry M.

    2017-01-01

    In late 2014, Zika virus (ZIKV; Flaviviridae, Flavivirus) emerged as a significant arboviral disease threat in the Western hemisphere. Aedes aegypti and Aedes albopictus have been considered the principal vectors of ZIKV in the New World due to viral isolation frequency and vector competence assessments. Limited reports of Culex transmission potential have highlighted the need for additional vector competence assessments of North American Culex species. Accordingly, North American Culex pipiens and Culex quinquefasciatus were orally exposed and intrathoracically inoculated with the African prototype ZIKV strain and currently circulating Asian lineage ZIKV strains to assess infection, dissemination, and transmission potential. Results indicated that these two North American Culex mosquito species were highly refractory to oral infection with no dissemination or transmission observed with any ZIKV strains assessed. Furthermore, both Culex mosquito species intrathoracically inoculated with either Asian or African lineage ZIKVs failed to expectorate virus in saliva. These in vivo results were further supported by the observation that multiple mosquito cell lines of Culex species origin demonstrated significant growth restriction of ZIKV strains compared with Aedes-derived cell lines. In summation, no evidence for the potential of Cx. pipiens or Cx. quinquefasciatus to serve as a competent vector for ZIKV transmission in North America was observed. PMID:28500817

  7. The cuticle proteins: a putative role for deltamethrin resistance in Culex pipiens pallens.

    PubMed

    Fang, Fujin; Wang, Weijie; Zhang, Donghui; Lv, Yuan; Zhou, Dan; Ma, Lei; Shen, Bo; Sun, Yan; Zhu, Changliang

    2015-12-01

    Insecticide resistance has been a major public health challenge. It is impendent to study the mechanism on insecticide resistance. In our previous study, 14 differentially accumulated insect cuticle proteins (ICPs) based on insecticide resistance proteomes and transcriptomes were found in the deltamethrin-resistant (DR) and -susceptible (DS) strains of Culex pipiens pallens. To investigate if these ICPs are associated with deltamethrin resistance, different transcriptional levels of the 14 ICPs were detected in the DS and DR strains from laboratory and field populations by using quantitative real-time polymerase chain reaction (qRT-PCR). The expression levels of the 14 ICPs were also measured after short-term exposure of the DS strain to deltamethrin. The full-length complementary DNA (cDNA) of CpCPLCG5 gene, which encodes one of the 14 ICPs, was cloned from Cx. pipiens pallens. Homology analysis and phylogenetic analysis were carried out with some other insects. Furthermore, small interfering RNA (siRNA) was used to knockdown the expression level of CpCPLCG5 gene for characterizing its contribution to deltamethrin resistance. The results showed that the expression level of CpCPLCG5 gene was higher in DR strain than in DS strain both in laboratory and field populations while the other 13 ICPs were downregulated. The full-length cDNA of CpCPLCG5 gene was 732 bp, with the ORF of 390 bp and deduced 129 amino acids (GenBank/KF723314,2013). Knockdown of CpCPLCG5 gene increased the susceptibility of the DR strain while the expression level of the other 13 ICPs elevated. Our findings indicate that the cuticle proteins are associated with deltamethrin resistance in Cx. pipiens pallens.

  8. 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.

  9. Evidence for facilitation of Culex pipiens (Diptera: Culicidae) life history traits by the nonnative invasive shrub Amur honeysuckle (Lonicera maackii).

    PubMed

    Shewhart, Lauren; McEwan, Ryan W; Benbow, M Eric

    2014-12-01

    Mosquitoes are one of the most globally important insect pests and vectors of human pathogens, and their populations may be facilitated or inhibited by anthropogenic environmental change. Invasive plant species are an important management concern and environmental modifier in many ecosystems; these plant invasions have the potential to exacerbate or diminish mosquito populations. The purpose of this study was to assess potential effects of a highly invasive plant, Lonicera maackii, on a common mosquito species Culex pipiens L., which is an important pathogen vector in the United States. Three microcosm assays were conducted to determine the responses of C. pipiens life history attributes of larval survivorship, growth, and pupation when subjected to leachate from two native plant leaves (Platanus occidentalis and Acer saccharum) and both the leaves and flowers of L. maackii. Only C. pipiens larvae exposed to L. maackii leachate pupated and emerged as adults. However, in all three assays there were statistically significant differences in survivorship and body size change among treatments, and in each assay the highest survivorship and maximum larval size was found in the L. maackii leachate treatments, suggesting positive effects on certain life history traits. This study is one of the first to demonstrate the potential facilitative effect of this invasive plant species on an insect vector and suggests that plant invasion could have positive feedbacks into mosquito population dynamics and, ultimately, human disease.

  10. The complete mitochondrial genome of the Rana huanrensis (Anura: Ranidae).

    PubMed

    Dong, Bingjun; Zhou, Yu; Yang, Baotian

    2016-11-01

    We first determined complete mitochondrial genomes of R. huanrensis (Anura: Ranidae). The complete mtDNA sequence is 19 253 bp in length, including 13 protein-coding genes, 2 rRNA genes, 22 tRNA genes, and one displacement loop. The start/stop codons of protein-coding genes are similar to which of R. chensinensis. D-loop region of R. huanrensis is 3448 bp in size, contains many tandem repeat units. The phylogenetic trees of 18 species from Ranidae were reconstructed by BI and ML analyses. The result indicated that R. huanrensis is the most closely related species with other Rana species. The molecular data are expected to provide a useful tool for population genetics studies of this species and further phylogenetic analyses of Ranidae.

  11. Temporal changes in mosquito abundance (Culex pipiens), avian malaria prevalence and lineage composition

    PubMed Central

    2013-01-01

    Background Knowledge on the temporal dynamics of host/vector/parasite interactions is a pre-requisite to further address relevant questions in the fields of epidemiology and evolutionary ecology of infectious diseases. In studies of avian malaria, the natural history of Plasmodium parasites with their natural mosquito vectors, however, is mostly unknown. Methods Using artificial water containers placed in the field, we monitored the relative abundance of parous females of Culex pipiens mosquitoes during two years (2010–2011), in a population in western Switzerland. Additionally, we used molecular tools to examine changes in avian malaria prevalence and Plasmodium lineage composition in female C. pipiens caught throughout one field season (April-August) in 2011. Results C. pipiens relative abundance varied both between years and months, and was associated with temperature fluctuations. Total Plasmodium prevalence was high and increased from spring to summer months (13.1-20.3%). The Plasmodium community was composed of seven different lineages including P. relictum (SGS1, GRW11 and PADOM02 lineages), P. vaughani (lineage SYAT05) and other Plasmodium spp. (AFTRU5, PADOM1, COLL1). The most prevalent lineages, P. vaughani (lineage SYAT05) and P. relictum (lineage SGS1), were consistently found between years, although they had antagonistic dominance patterns during the season survey. Conclusions Our results suggest that the time window of analysis is critical in evaluating changes in the community of avian malaria lineages infecting mosquitoes. The potential determinants of the observed changes as well as their implications for future prospects on avian malaria are discussed. PMID:24499594

  12. Immunohistochemical colocalization of 7B2 and 5HT in the neuroepithelial bodies of the lung of Rana temporaria.

    PubMed

    Bodegas, M E; Montuenga, L M; Polak, J M; Sesma, P

    1993-07-01

    The neuroendocrine cell population of the lung of Rana temporaria has been studied by means of immunocytochemistry. Serotonin (5HT)- and polypeptide 7B2-immunoreactive neuroepithelial bodies have been observed in the epithelial lining of the lung. 5HT- but not 7B2-immunoreactive isolated endocrine cells have also been observed.

  13. Postbreeding movements of the dark gopher frog, Rana sevosa goin and netting: Implications for conservation and management

    Treesearch

    Stephen C. Richter; Jeanne E. Young; Richard A. Seigal; Glen N. Johnson

    2001-01-01

    Conservation plans for amphibians often focus on activities at the breeding site, but for species that use temstrial habitats for much of the year, an understanding of nonbreeding habitat use is also essential. We used radio telemetry to study the postbreeding movements of individuals of the only known population of dark gopher frogs, Rana sevosa,...

  14. 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

  15. [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

  16. Energetic cost of insecticide resistance in Culex pipiens mosquitoes.

    PubMed

    Rivero, A; Magaud, A; Nicot, A; Vézilier, J

    2011-05-01

    The extensive use of insecticides to control vector populations has lead to the widespread development of different mechanisms of insecticide resistance. Mutations that confer insecticide resistance are often associated to fitness costs that prevent them from spreading to fixation. In vectors, such fitness costs include reductions in preimaginal survival, adult size, longevity, and fecundity. The most commonly invoked explanation for the nature of such pleiotropic effects of insecticide resistance is the existence of resource-based trade-offs. According to this hypothesis, insecticide resistance would deplete the energetic stores of vectors, reducing the energy available for other biological functions and generating trade-offs between insecticide resistance and key life history traits. Here we test this hypothesis by quantifying the energetic resources (lipids, glycogen, and glucose) of larvae and adult females of the mosquito Culex pipiens L. resistant to insecticides through two different mechanisms: esterase overproduction and acetylcholinesterase modification. We find that, as expected from trade-off theory, insecticide resistant mosquitoes through the overproduction of esterases contain on average 30% less energetic reserves than their susceptible counterparts. Acetylcholinesterase-modified mosquitoes, however, also showed a significant reduction in energetic resources (20% less). We suggest that, in acetylcholinesterase-modified mosquitoes, resource depletion may not be the result of resource-based trade-offs but a consequence of the hyperactivation of the nervous system. We argue that these results not only provide a mechanistic explanation for the negative pleiotropic effects of insecticide resistance on mosquito life history traits but also can have a direct effect on the development of parasites that depend on the vector's energetic reserves to fulfil their own metabolic needs.

  17. Long-term in vivo modulation of synaptic efficacy at the neuromuscular junction of Rana pipiens frogs

    PubMed Central

    Bélair, Eve-Lyne; Vallée, Joanne; Robitaille, Richard

    2005-01-01

    Prolonged changes in motor neurone activity can result in long-term changes in synaptic transmission. We investigated whether mechanisms commonly thought to be involved in determining synaptic efficacy of vertebrate motor neurones are involved in these long-term changes. The nerve supplying the cutaneus pectoris muscle was chronically stimulated via skin surface electrodes in freely moving frogs for 5–7 days. Chronic stimulation induced a 50% reduction in evoked endplate potential (EPP) amplitude at stimulated neuromuscular junctions (NMJs). These changes appear to be presynaptic since miniature EPP (mEPP) amplitude was unchanged while mEPP frequency was decreased by 46% and paired-pulse facilitation was increased by 26%. High frequency facilitation (40 Hz, 2 s) was also increased by 89%. Moreover, stimulated NMJs presented a 92% decrease in synaptic depression (40 Hz, 2 s). An increase in mitochondrial metabolism was observed as indicated by a more pronounced labelling of active mitochondria (Mitotracker) in stimulated nerve terminals, which could account for their greater resistance to synaptic depression. NMJ length visualized by α-bungarotoxin staining of nAChRs was not affected. Presynaptic calcium signals measured with Calcium Green-1 were larger in stimulated NMJs at low frequency (0.2 Hz) and not different from control NMJs at higher frequency (40 Hz, 2 s and 30 s). These results suggest that some mechanisms downstream of calcium entry are responsible for the determination of synaptic output, such as a down-regulation of some calcium-binding proteins, which could explain the observed results. The possibility of a change in frequenin expression, a calcium-binding protein that is more prominently expressed in phasic synapses, was, however, refuted by our results. PMID:16166159

  18. Regeneration of lumbar dorsal root axons into the spinal cord of adult frogs (Rana pipiens), an HRP study.

    PubMed

    Liuzzi, F J; Lasek, R J

    1985-02-22

    Lumbar dorsal roots of adult frogs were crushed or cut and reanastomosed. Following survival times of up to 75 days, the regenerating dorsal roots were recut and anterogradely injury-filled with horseradish peroxidase. This revealed that in the adult frog, regenerating axons re-enter the spinal cord. Comparison of the distribution of these axons with that of normal dorsal root axons showed that there is a partial restoration of the segmental distribution in the gray matter. However, the long ascending sensory tract of the dorsal funiculus was not restored. The dorsal funiculus was markedly gliotic and had relatively few labelled, regenerated axons. The labelled axons that were seen in the dorsal funiculus either extended longitudinally for a distance just beneath the pia, apparently in association with the glia limitans, or traversed the region to enter the dorsal gray matter. Most of the large and small diameter axons that entered the gray matter did so by passing through the region of the dorsolateral fasciculus. Within the gray matter, small diameter, regenerated axons arborized in the region of the dorsal terminal field, a region that has been shown in the normal frog to receive cutaneous afferents only. Many large diameter axons, presumably muscle afferents, arborized in the ventral terminal field, a region shown in the normal frog to receive muscle afferents exclusively. However, many of these large diameter axons had arborizations that extended to both terminal fields, thus suggesting that some abberant connections are made during dorsal root regeneration in the adult frog.

  19. 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 ...

  20. 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 ...

  1. Superimposed maps of the monocular visual fields in the caudolateral optic tectum in the frog, Rana pipiens.

    PubMed

    Winkowski, Daniel E; Gruberg, Edward R

    2005-01-01

    The superficial layers of the frog optic tectum receive a projection from the contralateral eye that forms a point-to-point map of the visual field. The monocular part of the visual field of the contralateral eye is represented in the caudolateral region of the tectum while the binocular part of the visual field is represented in the rostromedial tectum. Within the representation of the binocular field (rostromedial tectum), the maps of visual space from each eye are aligned. The tectal representation of the binocular visual field of the ipsilateral eye is mediated through a crossed projection from the midbrain nucleus isthmi. This isthmotectal projection also terminates in the caudolateral region of the optic tectum, yet there has been no indication that it forms a functional connection. By extracellular recording in intermediate layer 7 of the caudolateral tectum, we have discovered electrical activity driven by visual stimulation in the monocular visual field of the ipsilateral eye. The units driven from the ipsilateral eye burst upon initial presentation of the stimulus. At individual layer 7 recording sites in the caudolateral tectum, the multiunit receptive field evoked from the ipsilateral eye is located at the mirror image spatial location to the multiunit receptive field driven by the contralateral eye. Thus, as revealed electrophysiologically, there are superimposed topographic maps of the monocular visual fields in the caudolateral tectum. The ipsilateral eye monocular visual field representation can be abolished by electrolytic ablation of contralateral nucleus isthmi.

  2. 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

  3. Avian host-selection by Culex pipiens in experimental trials.

    PubMed

    Simpson, Jennifer E; Folsom-O'Keefe, Corrine M; Childs, James E; Simons, Leah E; Andreadis, Theodore G; Diuk-Wasser, Maria A

    2009-11-17

    Evidence from field studies suggests that Culex pipiens, the primary mosquito vector of West Nile virus (WNV) in the northeastern and north central United States, feeds preferentially on American robins (Turdus migratorius). To determine the contribution of innate preferences to observed preference patterns in the field, we conducted host preference trials with a known number of adult female C. pipiens in outdoor cages comparing the relative attractiveness of American robins with two common sympatric bird species, European starling, Sternus vulgaris and house sparrow, Passer domesticus. Host seeking C. pipiens were three times more likely to enter robin-baited traps when with the alternate host was a European starling (n = 4 trials; OR = 3.06; CI [1.42-6.46]) and almost twice more likely when the alternative was a house sparrow (n = 8 trials; OR = 1.80; CI = [1.22-2.90]). There was no difference in the probability of trap entry when two robins were offered (n = 8 trials). Logistic regression analysis determined that the age, sex and weight of the birds, the date of the trial, starting-time, temperature, humidity, wind-speed and age of the mosquitoes had no effect on the probability of a choosing a robin over an alternate bird. Findings indicate that preferential feeding by C. pipiens mosquitoes on certain avian hosts is likely to be inherent, and we discuss the implications innate host preferences may have on enzootic WNV transmission.

  4. Diversity and phylogeography of Northeast Asian brown frogs allied to Rana dybowskii (Anura, Ranidae).

    PubMed

    Yang, Bao-Tian; Zhou, Yu; Min, Mi-Sook; Matsui, Masafumi; Dong, Bing-Jun; Li, Pi-Peng; Fong, Jonathan J

    2017-07-01

    We investigated the species diversity and phylogeography of the Northeast Asian brown frogs allied to Rana dybowskii (the R. dybowskii species complex: R. dybowskii, R. pirica, and R. uenoi) using four mitochondrial and three nuclear loci. Phylogenetic analyses confirmed the existence of three distinct species in this complex; using extensive molecular data, we confirm the validity of Rana uenoi recognized as a distinct species, and infer R. dybowskii and R. pirica to be sister species. Also, we included populations from previously unsampled regions in Northeast China, and identified them to be R. dybowskii. While many species in Northeast Asia diverged due to Pleistocene glaciation, divergence-dating analyses inferred older, Miocene speciation in the R. dybowskii species complex. Ancestral area reconstruction identified the orogenic movement of the Changbai Mountain Range and the opening of the Sea of Japan/East Sea being major events influencing allopatric speciation. Copyright © 2017 Elsevier Inc. All rights reserved.

  5. Topological analysis of the brain stem of the frogs Rana esculenta and Rana catesbeiana.

    PubMed

    Opdam, R; Kemali, M; Nieuwenhuys, R

    1976-02-01

    The ventricular sulcal pattern and the cytoarchitectonic organization of the brain stem of the frogs Rana esculenta and Rana catesbeiana have been studied in transversely cut, Nissl stained serial sections. Four longitudinal sulci, the sulcus medianus inferior, the sulcus intermedius ventralis, the sulcus limitans and the sulcus medianus superior could be distinguished in both species. A fifth longitudinal groove, the sulcus intermedius dorsalis, was found only in Rana esculenta. With the aid of the usual cytoarchitectonic criteria 25 cell masses have been delineated in Rana esculenta and 27 in Rana catesbeiana. These cell masses can be distributed over the following categories (numbers added in brackets for Rana catesbeiana, if different from those in Rana esculenta): primary efferent or motor, 8; primary afferent or sensory, 4(6); "relay" centers, 7. Contrary to statements in the literature the reticular formation can be divided into six separate cell groups. The majority of the nuclei form part of the central gray, which constitutes a rather wide zone in anurans; three reticular nuclei lie partly within the stratum griseum and partly within the stratum album; six nuclei are entirely embedded in the stratum album. The morphological pattern of the cell masses and their relationship to the ventricular sulci were studied with the aid of a graphical reconstruction procedure termed topological analysis (cf. Nieuwenhuys, '74 and figs. 15, 16). This analysis yielded the following results: The sulcus limitans extends throughout the rhombencephalon, dividing this brain part into a basal plate and an alar plate. The cell masses in the basal plate fit into two longitudinal zones, a medial area ventralis and a lateral area intermedioventralis. The area ventralis contains three somatic motor nuclei (IV, VI and XII) and the rhombencephalic medial reticular zone. The latter may be primarily considered as a somatic motor coordinating center. The area intermedioventralis contains

  6. 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.

  7. Transcriptional Regulation of Culex pipiens Mosquitoes by Wolbachia Influences Cytoplasmic Incompatibility

    PubMed Central

    Harris, Simon; Kambris, Zakaria; Sutton, Elizabeth R.; Bonsall, Michael B.; Parkhill, Julian; Sinkins, Steven P.

    2013-01-01

    Cytoplasmic incompatibility (CI) induced by the endosymbiont Wolbachia pipientis causes complex patterns of crossing sterility between populations of the Culex pipiens group of mosquitoes. The molecular basis of the phenotype is yet to be defined. In order to investigate what host changes may underlie CI at the molecular level, we examined the transcription of a homolog of the Drosophila melanogaster gene grauzone that encodes a zinc finger protein and acts as a regulator of female meiosis, in which mutations can cause sterility. Upregulation was observed in Wolbachia-infected C. pipiens group individuals relative to Wolbachia-cured lines and the level of upregulation differed between lines that were reproductively incompatible. Knockdown analysis of this gene using RNAi showed an effect on hatch rates in a Wolbachia infected Culex molestus line. Furthermore, in later stages of development an effect on developmental progression in CI embryos occurs in bidirectionally incompatible crosses. The genome of a wPip Wolbachia strain variant from Culex molestus was sequenced and compared with the genome of a wPip variant with which it was incompatible. Three genes in inserted or deleted regions were newly identified in the C. molestus wPip genome, one of which is a transcriptional regulator labelled wtrM. When this gene was transfected into adult Culex mosquitoes, upregulation of the grauzone homolog was observed. These data suggest that Wolbachia-mediated regulation of host gene expression is a component of the mechanism of cytoplasmic incompatibility. PMID:24204251

  8. The Arrival of the Northern House Mosquito Culex pipiens (Diptera: Culicidae) on Newfoundland's Avalon Peninsula.

    PubMed

    Chaulk, Andrew C; Carson, Kate P; Whitney, Hugh G; Fonseca, Dina M; Chapman, Thomas W

    2016-11-01

    Culex pipiens L., the northern house mosquito, is the primary vector of West Nile virus to humans along the east coast of North America and thus the focus of much study. This species is an urban container-breeding mosquito whose close contact with humans and flexibility in host choice has led to its classification as a "bridge vector"; that is, it is thought to move zoonotic diseases to humans from vertebrate reservoirs. While this invasive species is now well documented in its established range, which expanded in 2001 to include Canada, the existence of populations of this species along the fringes of its range are less well known. Here we report, using morphological and genetic techniques, the existence of two locations where Cx. pipiens exists in Newfoundland in both expected and unexpected sites based on projected habitat suitability on the island. © The Authors 2016. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  9. Culex pipiens forms and urbanization: effects on blood feeding sources and transmission of avian Plasmodium.

    PubMed

    Martínez-de la Puente, Josué; Ferraguti, Martina; Ruiz, Santiago; Roiz, David; Soriguer, Ramón C; Figuerola, Jordi

    2016-12-08

    The wide spread mosquito Culex pipiens pipiens have two forms molestus and pipiens which frequently hybridize. The two forms have behavioural and physiological differences affecting habitat requirements and host selection, which may affect the transmission dynamic of Cx. p. pipiens-borne diseases. During 2013, blood engorged Cx. p. pipiens mosquitoes were captured in urban, rural and natural areas from Southern Spain. In 120 mosquitoes, we identified the blood meal origin at vertebrate species/genus level and the mosquito form. The presence and molecular lineage identity of avian malaria parasites in the head-thorax of each mosquito was also analysed. Mosquitoes of the form pipiens were more frequently found in natural than in urban areas. The proportion of Cx. pipiens form molestus and hybrids of the two forms did not differ between habitat categories. Any significant difference in the proportion of blood meals on birds between forms was found. Birds were the most common feeding source for the two forms and their hybrids. Among mammals, dogs and humans were the most common hosts. Two Plasmodium and one Haemoproteus lineages were found in mosquitoes, with non-significant differences between forms. This study supports a differential distribution of Cx. p. pipiens form pipiens between urban and natural areas. Probably due to the similar feeding sources of both mosquito forms and their hybrids here, all of them may frequently interact with avian malaria parasites playing a role in the transmission of Plasmodium.

  10. Ecological and genetic analyses of the complete genomes of Culex flavivirus strains isolated from Culex tritaeniorhynchus and Culex pipiens (Diptera: Culicidae) group mosquitoes.

    PubMed

    Obara-Nagoya, M; Yamauchi, T; Watanabe, M; Hasegawa, S; Iwai-Itamochi, M; Horimoto, E; Takizawa, T; Takashima, I; Kariwa, H

    2013-03-01

    Culex flavivirus (CxFV) is an insect-specific flavivirus that was first reported in 2007 in Japan. CxFV strains were isolated from Culex tritaeniorhynchus Giles and Culex pipiens L. group mosquitoes and genetically characterized in Toyama Prefecture, Japan, from 2004 to 2009, to reveal host specificity, mode of transmission, and seasonal and geographical distribution. The minimum infection rate (MIR) of CxFV within Cx. tritaeniorhynchus populations was 0.3 and much lower than that within Cx. pipiens group (17.9). The complete genome sequences of 11 CxFV isolates (four from Cx. tritaeniorhynchus and seven from Cx. pipiens group) consisted of 10,835-10,837 nucleotides. When these 11 isolates and five reference strains (NIID-21-2 and Tokyo strains from Japan, Iowa07 and HOU24518 strains from the United States, H0901 strain from China) were compared, there were 95.2-99.2% nucleotide and 98.1-99.8% amino acid identities. Phylogenetic analysis showed that the 11 isolates were divided into four clusters. One cluster consisted of five isolates from Cx. pipiens group and Cx. tritaeniorhynchus from one site and their nucleotide sequences almost completely matched. One cluster consisted of an isolate with a unique sequence from a Cx. pipiens group mosquito captured in an aircraft from Taiwan, suggesting that it was introduced from abroad. CxFV strains were divided into several groups according to countries when nucleotide sequences of CxFV available in GenBank and 11 Toyama isolates were compared. These results suggest that CxFV is maintained in nature among Culex mosquitoes in a mosquito habitat-specific but not a species-specific manner.

  11. Avian Host-Selection by Culex pipiens in Experimental Trials

    PubMed Central

    Simpson, Jennifer E.; Folsom-O'Keefe, Corrine M.; Childs, James E.; Simons, Leah E.; Andreadis, Theodore G.; Diuk-Wasser, Maria A.

    2009-01-01

    Evidence from field studies suggests that Culex pipiens, the primary mosquito vector of West Nile virus (WNV) in the northeastern and north central United States, feeds preferentially on American robins (Turdus migratorius). To determine the contribution of innate preferences to observed preference patterns in the field, we conducted host preference trials with a known number of adult female C. pipiens in outdoor cages comparing the relative attractiveness of American robins with two common sympatric bird species, European starling, Sternus vulgaris and house sparrow, Passer domesticus. Host seeking C. pipiens were three times more likely to enter robin-baited traps when with the alternate host was a European starling (n = 4 trials; OR = 3.06; CI [1.42–6.46]) and almost twice more likely when the alternative was a house sparrow (n = 8 trials; OR = 1.80; CI = [1.22–2.90]). There was no difference in the probability of trap entry when two robins were offered (n = 8 trials). Logistic regression analysis determined that the age, sex and weight of the birds, the date of the trial, starting-time, temperature, humidity, wind-speed and age of the mosquitoes had no effect on the probability of a choosing a robin over an alternate bird. Findings indicate that preferential feeding by C. pipiens mosquitoes on certain avian hosts is likely to be inherent, and we discuss the implications innate host preferences may have on enzootic WNV transmission. PMID:19924251

  12. 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.

  13. High genetic diversity in the Culex pipiens complex from a West Nile Virus epidemic area in Southern Europe.

    PubMed

    Simonato, Mauro; Martinez-Sañudo, Isabel; Cavaletto, Giacomo; Santoiemma, Giacomo; Saltarin, Andrea; Mazzon, Luca

    2016-03-15

    The Culex pipiens complex includes the most widespread mosquito species in the world. Cx. pipiens is the primary vector of the West Nile Virus (WNV) in Europe and North America. Cases of WNV have been recorded in Italy since 1998. In particular, wet areas along the Po River are considered some of the most WNV affected areas in Italy. Here, we analyzed the genetic structure of ten Cx. pipiens populations collected in the last part of the Po River including the Delta area. We assessed the genetic variability of two mitochondrial markers, cytochrome oxidase 1 (COI) and 2 (COII), for a total of 1200 bp, and one nuclear marker, a fragment of acetylcholinesterase-2 (ace-2), 502 bp long. The effect of the landscape features was evaluated comparing haplotype and nucleotide diversity with the landscape composition. The analysis showed a high genetic diversity in both COI and COII gene fragments mainly shared by the populations in the Delta area. The COI-COII network showed that the set of haplotypes found was grouped into three main supported lineages with the higher genetic variability gathered in two of the three lineages. By contrast, ace-2 fragment did not show the same differentiation, displaying alleles grouped in a single clade. Finally, a positive correlation between mitochondrial diversity and natural wetland areas was found. The high mitochondrial genetic diversity found in Cx. pipiens populations from the Po River Delta contrasts with the low variability of inland populations. The different patterns of genetic diversity found comparing mitochondrial and nuclear markers could be explained by factors such as differences in effective population size between markers, sex biased dispersal or lower fitness of dispersing females. Moreover, the correlation between genetic diversity and wetland areas is consistent with ecosystem stability and lack of insecticide pressure characteristic of this habitat. The mtDNA polymorphism found in the Po River Delta is even more

  14. 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

  15. 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.

  16. Sequence analysis of mtDNA COI barcode region revealed three haplotypes within Culex pipiens assemblage.

    PubMed

    Koosha, Mona; Oshaghi, Mohammad Ali; Sedaghat, Mohammad Mehdi; Vatandoost, Hassan; Azari-Hamidian, Shahyad; Abai, Mohammad Reza; Hanafi-Bojd, Ahmad Ali; Mohtarami, Fatemeh

    2017-10-01

    Members of the Culex (Culex) pipiens assemblage are known vectors of deadly encephalitides, periodic filariasis, and West Nile virus throughout the world. However, members of this assemblage are morphologically indistinguishable or hard to distinguish and play distinct roles in transmission of the diseases. The current study aimed to provide further evidence on utility of the two most popular nuclear (ITS2-rDNA) and mitochondrial (COI barcode region) genetic markers to identify members of the assemblage. Culex pipiens assemblage specimens from different climate zones of Iran were collected and identified to species level based on morphological characteristics. Nucleotide sequences of the loci for the specimens plus available data in the GenBank were analyzed to find species specific genetic structures useful for diagnosis purposes. ITS2 region was highly divergent within species or populations suggesting lack of consistency as a reliable molecular marker. In contrast, sequence analysis of 710 bp of COI gene revealed three fixed haplotypes named here "C, T, H" within the assemblage which can be distinguished by HaeIII and AluI enzymes. There were a correlation between the haplotypes and the world climate regions, where the haplotypes H/T and C are present mainly in temperate and tropical regions of the world, respectively. In the New world, Australia, and Japan only haplotype H is found. In conjunction between tropical and temperate regions such Iran, China, and Turkey, a mix of C/H or C/H/T are present. Although, the haplotypes are not strictly species-specific, however, Cx. quinquefasciatus was mainly of haplotype C. Due to the lack of mating barrier and questionable taxonomic situation of the complex members, the mentioned haplotypes in combination with other morphological and molecular characters might be used to address the genetic structure of the studied populations. Copyright © 2017 Elsevier Inc. All rights reserved.

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

    PubMed Central

    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

    2015-01-01

    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 1,826 significantly differentially expressed genes (DEGs). Among them, 1,078 genes were up-regulated and 748 genes were down-regulated in the deltamethrin-resistant strain compared to -susceptible strain. These DEGs contained cytochrome P450s, cuticle proteins, UDP-glucuronosyltransferases, lipases, serine proteases, heat shock proteins, esterases and others. Among the 1,826 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

  18. Helminths of the two mountain frogs, banded frog, Rana camerani Boulenger, 1886 and Uludağ frog Rana macrocnemis Boulenger, 1885 (Anura: Ranidae), collected from the Antalya province.

    PubMed

    Düşen, Serdar

    2007-01-01

    In this study, two mountain frogs (Rana camerani and Rana macrocnemis) were collected in the Antalya Province in south-western Turkey during 2001 and 2002 and were examined for helminths. Out of 15 Rana camerani, 10 (66.7%) were infected with 1 or more helminths and out of 20 Rana macrocnemis, 17 (85%) were infected with 1 or more helminths. The helminth fauna of Rana camerani included 4 species of which were 3 trematode species (Haplometra cylindracea, Pleurogenoides medians, Opisthioglyphe rastellus), and 1 nematode species (Cosmocerca ornata). The helminth fauna of Rana macrocnemis included 3 species with 1 trematode species (H. cylindracea), 1 nematode species (C. ornata), and 1 acanthocephalan species (Acanthocephalus ranae). H. cylindracea and C. ornata were observed in both of the mountain frogs.

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

    USDA-ARS?s Scientific Manuscript database

    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...

  20. Asymmetrical Competition and Patterns of Abundance of Aedes albopictus and Culex pipiens (Diptera: Culicidae)

    PubMed Central

    COSTANZO, KATIE S.; MORMANN, KIMBERLY; JULIANO, STEVEN A.

    2007-01-01

    We tested for competitive advantage among larvae of Aedes albopictus (Skuse) and Culex pipiens L. in a laboratory experiment and determined the frequency and spatial and temporal patterns of co-occurrence in the field in East St. Louis, IL. In a laboratory competition experiment at multiple combined densities of Ae. albopictus and Cx. pipiens larvae, Ae. albopictus survivorship and developmental times were significantly affected by conspecific densities but not by Cx. pipiens densities. In contrast, Cx. pipiens survivorship and developmental times were significantly affected by both conspecific and Ae. albopictus densities. Per capita rate of increase (r′) for Ae. albopictus cohorts declined significantly due to density of conspecifics, but not density of Cx. pipiens. Interspecific competition between Ae. albopictus and Cx. pipiens under these laboratory conditions was strong and asymmetrical, with the effect of Ae. albopictus on Cx. pipiens much stronger than the reverse. In monthly samples from tire sites in East St. Louis, Ae. albopictus was highly seasonal, occurring in relatively low abundance from early May to July and increasing in abundance in August and September. Co-occurrence corresponded to the seasonality of Ae. albopictus, with Cx. pipiens encountering Ae. albopictus in more tires and at higher numbers within a tire, in August and September. Abundance of both species was high in residential areas and was unrelated to overstory cover, total nitrogen, and total phosphorus. Abundance of Ae. albopictus, but not of Cx. pipiens, was positively associated with conductivity. We expect Cx. pipiens to suffer from the effects of interspecific competition in tires in which it encounters Ae. albopictus. Interspecific competition between these species may be of both ecological and medical importance. PMID:16119544

  1. Eight new tetranucleotide microsatellite loci for the agile frog (Rana dalmatina).

    PubMed

    Hauswaldt, J S; Fuessel, J; Guenther, J; Steinfartz, S

    2008-11-01

    We describe eight new polymorphic tetranucleotide microsatellite loci isolated from the agile frog (Rana dalmatina). In 25 individuals from the Nature Reserve Lüneburger Heide (Lower Saxony, Germany), the number of alleles per locus ranged from four to nine and average observed heterozygosities from 69.1% to 80.7%. No evidence for linkage disequilibrium was found and none of the loci showed significant deviation from Hardy-Weinberg expectations. These microsatellite DNA markers are suitable tools for addressing population genetics issues in this endangered species.

  2. Bionomics and control of Culex pipiens fatigans Wied. in Ceylon

    PubMed Central

    Chow, C. Y.; Thevasagayam, E. S.

    1957-01-01

    The climatic and housing conditions which favour the breeding of Culex pipiens fatigans in Ceylon, and its life-cycle and resting-habits, are described. The results of the treatment of breeding-places (mainly catch-pits) and the interior of houses with various insecticides of specific action on larvae or adults, in different preparations and concentrations, are given, and suggestions are made for the control of the mosquito on this island. The fundamental measure for successful control is efficient sanitation, but until this can be achieved, application of larvicides seems to be the method of choice. PMID:13472415

  3. Evidence from the primary structures of dermal antimicrobial peptides that Rana tagoi okiensis and Rana tagoi tagoi (Ranidae) are not conspecific subspecies.

    PubMed

    Conlon, J Michael; Coquet, Laurent; Jouenne, Thierry; Leprince, Jérôme; Vaudry, Hubert; Iwamuro, Shawichi

    2010-01-01

    Morphological evidence and data from comparisons of nucleotide sequences of mitochondrial genes demonstrate considerable intraspecies variation among populations of the Japanese brown frog Rana tagoi Okada 1928 (Tago's brown frog). Five peptides with antimicrobial activity were isolated from an extract of the skins of specimens of Rana tagoi okiensis collected on the Oki Islands, Japan. Determination of their primary structures demonstrated that two peptides belong to the ranatuerin-2 family, two peptides to the temporin family, and one peptide to the brevinin-1 family. Ranatuerin-2 peptides were not previously identified in the skin of specimens of R. t. tagoi collected in Chiba Prefecture, Japan and the structures of the temporin peptides from R. t. okiensis (temporin-TOa: FLPILGKLLSGFL.NH(2) and temporin-TOb: FLPILGKLLSGLL.NH(2)) are different from temporin-TGa (FLPILGKLLSGIL.NH(2)) isolated from R. t. tagoi. Similarly, the acyclic C-terminally alpha-amidated brevinin-1 peptide from R. t. okiensis (Brevinin-1TOa, GIGSILGVIAKGLPTLISWIKNR.NH(2)) shows three amino acid substitutions (Gly(1)-->Ala, Val(8)-->Ala, Ile(9)-->Leu) compared to the ortholog from R. t. tagoi. In addition, bradykinin, identical to the mammalian peptide, is present in high concentration in the skin of R. t. okiensis but not R. t. tagoi. The data provide evidence to support the proposal that R. t. tagoi and R. t. okiensis should be regarded as separate species (R. tagoi and R. okiensis) rather than conspecific subspecies. Copyright 2009 Elsevier Ltd. All rights reserved.

  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. Culex pipiens, an experimental efficient vector of West Nile and Rift Valley fever viruses in the Maghreb region.

    PubMed

    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 10(7.8) and 10(8.5) 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.

  6. The effects of forced-egg retention on the blood-feeding behavior and reproductive potential of Culex pipiens (Diptera: Culicidae).

    PubMed

    Johnson, Brian J; Fonseca, Dina M

    2014-07-01

    High rates of West Nile virus (WNV) transmission to humans are associated with exceptionally hot and dry summers. This is paradoxical since the eggs of Culex vectors of WNV depend on the persistence of containers with water, which decline during droughts. We examined the effects of forced-egg retention on the reproductive success of female Culex pipiens as well as behavioral responses, such as likelihood of secondary blood meals. As controls we examined the effects of female age and delayed mating. We found that early mating is essential to achieve reproductive success and, consistent with an "all-or-none" ovipositing strategy, C. pipiens females are able to retain considerable reproductive potential while searching for oviposition sites. Specifically, although forced-egg retention resulted in significant decreases in fitness, the decline was moderate for 5 weeks and most can be accounted for by increases in female age. Consequently, no females took blood more than once per gonotrophic cycle, which eliminates the possibility that heightened vectorial capacity due to multiple blood-feedings increases WNV transmission during periods of drought. Instead, our findings suggest that during droughts populations of C. pipiens have time to locate the remaining water holes, which are associated with human populations and WNV-competent bird species.

  7. Plasmodium relictum (lineages pSGS1 and pGRW11): complete synchronous sporogony in mosquitoes Culex pipiens pipiens.

    PubMed

    Kazlauskienė, Rita; Bernotienė, Rasa; Palinauskas, Vaidas; Iezhova, Tatjana A; Valkiūnas, Gediminas

    2013-04-01

    Plasmodium relictum is a widespread invasive agent of avian malaria, responsible for acute, chronic and debilitating diseases in many species of birds. Recent PCR-based studies revealed astonishing genetic diversity of avian malaria parasites (genus Plasmodium), with numerous genetic lineages deposited in GenBank. Many studies addressed distribution and evolutionary relationships of avian Plasmodium lineages, but information about patterns of development of different lineages in mosquito vectors remains insufficient. Here we present data on sporogonic development of 2 widespread mitochondrial cytochrome b lineages (cyt b) of P. relictum (pSGS1 and pGRW11) in mosquito Culex pipiens pipiens. Genetic distance between these lineages is 0.2%; they fall in a well-supported clade in the phylogenetic tree. Three P. relictum strains were isolated from common crossbill (Loxia curvirostra, lineage pSGS1), domestic canary (Serinus canaria domestica, pSGS1) and house sparrow (Passer domesticus, pGRW11). These strains were multiplied in domestic canaries and used as donors of malarial gametocytes to infect C. p. pipiens. Mosquitoes were allowed to take blood meal on infected canaries and then dissected on intervals to study development of sporogonic stages. All 3 strains developed synchronously and completed sporogony in this vector, with infective sporozoites reported in the salivary glands on the day 14 after infection. Ookinetes, oocysts and sporozoites of all strains were indistinguishable morphologically. This study shows that patterns of sporogonic development of the closely related lineages pSGS1 and pGRW11 and different strains of the lineage pSGS1 of P. relictum are similar indicating that phylogenetic trees based on the cyt b gene likely can be used for predicting sporogonic development of genetically similar avian malaria lineages in mosquito vectors. Copyright © 2013 Elsevier Inc. All rights reserved.

  8. 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

    Conley, Amy K; Fuller, Douglas O; Haddad, Nabil; Hassan, Ali N; Gad, Adel M; Beier, John C

    2014-06-24

    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. 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. 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 independent surveys or previous disease outbreaks. Cx

  9. 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

  10. The molecular and morphological variations of Culex pipiens complex (Diptera: Culicidae) in Iran.

    PubMed

    Dehghan, H; Sadraei, J; Moosa-Kazemi, S H; Baniani, N Akbari; Nowruzi, F

    2013-01-01

    Taxonomic status of Culex pipiens is well-known as many years with such a wide variety of morphological and biological characteristics. These changes have been the subject of extensive investigation by many researchers. There are a little information about the morphology and molecular data of Cx. pipiens complex in Iran. The taxonomic status of the complex is very important because of medical and veterinary importance and wide distribution in the country. This study was carried out in 11 areas in Iran using dipping technique from April 2009 to October 2010. Molecular study was carried out using primers F1457 as forward and B1256 as reverse, which amplified Ace.2 gene and performed PCR-RFLP using ScaI restriction enzyme. Culex quinquefasciatus found in south to central areas of Iran and reported as sympatric with Cx. pipiens in the central regions. Culex pipiens distributed in many areas of the country. Sequencing alignment of Ace.2 gene of Cx. quinquefasciatus and Cx. pipiens showed 6.5% variation in 46bp, especially in intron locus of gene. Culex pipiens complex from Iran are located in two separate clades with sister branches using phylogenetic sequencing tree. The male genitalia found as the most reliable diagnostic characters for identification of Cx. pipiens complex in Iran that confirmed by amplify the Ace.2 gene in the samples but we recommended the use of sequencing PCR products of microsatellite loci and COI gene in future study.

  11. Lectin Activity in Gut Extract of Culex pipiens

    PubMed Central

    Koosha, Mona; Abai, Mohammad Reza; Abolhasani, Mandan; Charedar, Soroor; Basseri, Hamid Reza

    2013-01-01

    Background: The role of lectins is important in interaction between pathogens and mosquito vectors. This study was performed to identify agglutinin activities of protein molecules on the midgut of Culex pipiens. Methods: Culex pipiens was reared in insectray condition and the midguts of males and females (blood fed and unfed) were dissected separately in Tris-HCl buffer. The extracts of midguts were applied for hemagglutinin assay against red blood cells of rabbit, mouse, rat, dog, horse, sheep, guinea pig, cow, human (A, B, AB, O groups). Then, the RBCs with relatively high agglutinin activity were chosen for carbohydrate inhibition assay. D (+) glucose, D (+) galactose, D (+) mannose, D (−) fructose, D (−) arabinose, L (−) fucose, lactose, N-acetyl-D-glucosamine, N-acetyl-D-galactosamine, sialic acid were used to specify carbohydrate binding lectin. Results: The highest agglutinin activities were found against sheep and rabbits RBCs. Sexual diversity of agglutinin activities was observed among midgut extraction of males and females. In addition, variation in agglutinin activity of blood fed and unfed female mosquitoes were detected. The lectin activity was inhibited highly with glucose, galactose, fucose and fructose but less inhibitor activities was observed by arabinose, N-acetyl-D-galactosamine, n-acetyl-d-glucosamine, lactose and mannose. Conclusion: The secretion of hemagglutinins (lectins or lectin-like molecules) in the digestive system depends on the type of food in the gut. This suggests that emptying of the gut in preparation for protein rich food probably starts the secretion of hemagglutinins. PMID:23785692

  12. 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.

  13. 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.

  14. 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?|

  15. Description of a new brown frog from Tsushima Island, Japan (Anura: Ranidae: Rana).

    PubMed

    Matsui, Masafumi

    2014-09-01

    Because all available evidence from allozymes, mtDNA sequences, and artificial hybridization suggests presence of high genetic differentiation between populations of East Asian brown frogs currently assigned to Rana dybowskii Günther, 1876, I compared morphological characters between specimens from Tsushima Island of Japan and Maritime territory of Russia. The population from Tsushima is slightly, but significantly different from R. dybowskii from Russia, including the holotype. I therefore consider the Tsushima population to be specifically distinct, and describe it as a new species R. uenoi. The new species also occurs in the Korean Peninsula and adjacent islands, but the distributional relationships with R. dybowskii are unclear, as detailed distribution in northern Korea is lacking.

  16. [Presumptive role of Wolbachia in deltamethrin resistance of Culex pipiens pallens].

    PubMed

    Hong, Shan-Chao; Lv, Yuan; Fang, Fu-Jin; Zhu, Chang-Liang

    2014-12-01

    To investigate the relationship between Wolbachia and deltamethrin resistance in Culex pipiens pallens. PCR was used to detect Wolbachia in Culex pipiens pallens and qRT-PCR was performed to determine and compare the expression of Wolbachia between deltamethrin- resistant and - susceptible strains of Culex pipiens pallens. Wolbachia was detected in Culex pipiens pallens in the laboratory. The expression of Wolbachia was 18.42, 3.69, 4.43, 3.96, 6.31, 1.55 and 3.76 folds higher in the deltamethrin-resistant strain than in susceptible strain in the egg, 1st, 2nd, 3rd, 4th stages, and male and female adults, but there was no statistical difference in the pupae stage. The expression of Wolbachia was 2.64 folds higher in deltamethrin-resistant females than in susceptible females which were caught in Jiangxinzhou of Nanjing. Wolbachia is associated with deltamethrin resistance in Culex pipines pallens.

  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.

  18. [The correlation of the ecological niches of the common (Rana temporaria L.) and of the moor (Rana arvalis Nilss.) frogs (Anura, Amphibia)].

    PubMed

    Severtsov, A S; Liapkov, S M; Surova, G S

    1998-01-01

    During 25 years ecology and population dynamic of two brown frog species (Rana temporaria and R. arvalis) were studied in Moscow region, Solovki island and South Ural. We compared life cycles characteristics, namely biotope preferences, diet, migration, enemies, hibernation places using own and available literature data. Then we analyse how these parameters are changed among the species area and ecological niches were compared. We found that these two species do not compete in any stage of life cycle. Ecological niches are very closed and differences are determined generally by abiotic factors. So, R. temporaria prefers more wet biotope and more sensitive to acidity (low pH value). Differences in spawning time do not associate with interference in spawning places. We conclude that interspecific competition did not take place neither in the past nor in present and the reason of differences in ecological niches are determined by separate ways of evolutionary development of these species.

  19. Patterns of infection by lungworms, Rhabdias ranae and Haematoloechus spp., in northern leopard frogs: a relationship between sex and parasitism.

    PubMed

    Dare, Oluwayemisi K; Forbes, Mark R

    2009-04-01

    We examined a population of northern leopard frogs to determine whether sex biases in investment in immunity, previously reported for this host species under controlled exposures to lung nematodes, is predictive of patterns of parasitism in nature. We examined Rhabdias ranae and Haematoloechus spp. infections in 74 breeding adult, 28 non-breeding adult, and 53 juvenile frogs. Contrary to our predictions, R. ranae prevalence and mean abundance were higher in breeding female frogs (prevalence: 39.4%, abundance: 3.05 +/- 0.85) than on breeding males (prevalence: 26.0%, abundance: 1.17 +/- 0.52), although no sex bias was observed among non-breeding adults or juvenile frogs. Female frogs also carried larger R. ranae worms, on average, than did males (females: 6407.38 microm +/- 153.80; males: 5198 microm +/- 131.09), regardless of age or breeding condition. We observed no sex-linked patterns of parasitism by Haematoloechus spp. worms in either adult or juvenile frogs. Alternative hypotheses, such as differences among sexes in the selection of thermal clines for hibernation, may explain the observed female bias in parasitism by nematode lungworms in nature and, thus, need to be considered.

  20. 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.

  1. 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.

  2. Dieldrin-resistance in Culex pipiens fatigans in Malaya

    PubMed Central

    Wharton, R. H.

    1958-01-01

    Resistance to insecticides in Culex pipiens fatigans has already been reported from two areas in Malaya. In Penang two years' use of BHC as a larvicide resulted in the development of a strain which was found to have acquired a tenfold resistance to BHC, and also to dieldrin to which it had not been exposed. In Singapore, when larval control became unsatisfactory after 6 months' use of a dieldrin emulsion, laboratory experiments confirmed that active resistance to dieldrin had developed. The present observations report the finding of two further dieldrin-BHC resistant strains of C. p. fatigans in Malaya, but differ from the previous reports in that resistance, in one strain at least, was developed as a result of house-spraying with dieldrin against adult mosquitos. In this strain resistance to dieldrin was about 100 times in both adults and larvae, resistance to gamma-BHC in larvae was about 20 times, while resistance to DDT was slight. PMID:13536808

  3. The susceptibility of Culex pipiens fatigans to residual insecticides*

    PubMed Central

    Smith, A.; Bransby-Williams, W. R.

    1962-01-01

    Observations have been made on Culex pipiens fatigans in the Taveta-Pare area in East Africa from 1954 to 1961, during which time dieldrin was applied to local houses over a 3½-year period as part of an experiment in malaria control. At the end of the period of residual spraying the numbers of C. p. fatigans had been reduced by two-thirds in the Taveta area, but in the South Pare area, only some 160 km away, this mosquito was twice as numerous as before spraying. The results of susceptibility tests carried out in untreated and dieldrin-treated areas showed that the susceptibility of C. p. fatigans in East Africa to chlorinated hydrocarbon insecticides was low compared with that of Anopheles gambiae, and varied by at least 20 times in some areas relatively short distances apart. C. p. fatigans from all areas sampled were, however, susceptible to fenthion and malathion. PMID:13993106

  4. Identification of proteins associated with pyrethroid resistance by iTRAQ-based quantitative proteomic analysis in Culex pipiens pallens.

    PubMed

    Wang, Weijie; Lv, Yuan; Fang, Fujin; Hong, Shanchao; Guo, Qin; Hu, Shengli; Zou, Feifei; Shi, Linna; Lei, Zhentao; Ma, Kai; Zhou, Dan; Zhang, Donghui; Sun, Yan; Ma, Lei; Shen, Bo; Zhu, Changliang

    2015-02-10

    Mosquito control based on chemical insecticides is considered as an important element in the current global strategies for the control of mosquito-borne diseases. Unfortunately, the development of pyrethroid resistance in important vector mosquito species jeopardizes the effectiveness of insecticide-based mosquito control. To date, the mechanisms of pyrethroid resistance are still unclear. Recent advances in proteomic techniques can facilitate to identify pyrethroid resistance-associated proteins at a large-scale for improving our understanding of resistance mechanisms, and more importantly, for seeking some genetic markers used for monitoring and predicting the development of resistance. We performed a quantitative proteomic analysis between a deltamethrin-susceptible strain and a deltamethrin-resistant strain of laboratory population of Culex pipiens pallens using isobaric tags for relative and absolute quantitation (iTRAQ) analysis. Gene Ontology (GO) analysis was used to find the relative processes that these differentially expressed proteins were involved in. One differentially expressed protein was chosen to confirm by Western blot in the laboratory and field populations of Cx. pipiens pallens. We identified 30 differentially expressed proteins assigned into 10 different categories, including oxidoreductase activity, transporter activity, catalytic activity, structural constituent of cuticle and hypothetical proteins. GO analysis revealed that 25 proteins were sub-categorized into 35 hierarchically-structured GO classifications. Western blot results showed that CYP6AA9 as one of the up-regulated proteins was confirmed to be overexpressed in the deltamethrin-resistant strains compared with the deltamethrin-susceptible strains both in the laboratory and field populations. This is the first study to use modern proteomic tools for identifying pyrethroid resistance-related proteins in Cx. pipiens. The present study brought to light many proteins that were not

  5. 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

  6. Interruption of chemical mosquito control and evolution of insecticide resistance genes in Culex pipiens (Diptera: Culicidae).

    PubMed

    Eritja, R; Chevillon, C

    1999-01-01

    Within the Llobregat Delta (Barcelona, Spain), Culex pipiens L. has been the target of organophosphate insecticide (OP) control for 10 yr (1982-1992). Between 1991 and 1992, OPs were replaced by Bacillus-based toxins in all the mosquito control programs within > 150 km of this area. The distribution of several OP-resistance genes was surveyed within the Llobregat Delta and neighboring populations (< 25 km) during the 2 yr following this regional pesticide change to investigate how the change in selection pressure affected the dynamics of OP-resistance genes. The immigration failure of the A2-B2 resistant esterases and the observed difference in OP-resistance dynamics between isolated and nonisolated populations may indicate fitness disadvantages associated with OP-resistance genes, hence a tendency for a decrease in OP-resistance. In contrast, one OP-resistance gene further increased in frequency, whereas the frequencies of some others were maintained. These unexpected results question the importance of pesticides from sources other than mosquito control, and the variability of pleiotropic fitness costs among pesticide resistance genes.

  7. Geographic distribution and relative abundance of the sibling vector species Culex torrentium and Culex pipiens in Sweden.

    PubMed

    Hesson, Jenny C; Ostman, Orjan; Schäfer, Martina; Lundström, Jan O

    2011-10-01

    Culex torrentium and Culex pipiens are sibling species and potential viral vectors that coexist in Europe. Larvae and females of the two species are morphologically almost identical, and reliable identification can only be done on males. To investigate the distribution and relative abundance of the two species in Sweden, we collected Culex larvae from sites spread over the country, identified them as Culex pipiens/torrentium based on morphology, and identified them to species using a recently developed restriction enzyme method. Cx. torrentium was the dominant species (89%, n=1012) and it occurred in 48 of the 49 sites investigated, and also dominated in most of the study sites. The proportion of Cx. pipiens larvae in relation to Cx. torrentium collected at each site decreased with both increasing latitude and altitude, and the presence of Cx. pipiens decreased with latitude. In addition, Cx. pipiens/torrentium females were sampled with Centres for Disease Control light traps baited with carbon dioxide. The overall country mean was 4.0 Cx. pipiens/torrentium caught per trap night, with decreasing numbers of Cx. pipiens/torrentium caught per trap night with increasing latitude. Thus, the abundance of Cx. pipiens/torrentium decreased, but the proportion Cx. torrentium increased, with increasing latitude. This is the first study that shows the vast dominance of Cx. torrentium over Cx. pipiens in Sweden. The unexpected dominance of Cx. torrentium highlights the importance of distinguishing between the two species in studies of Culex-borne arboviruses in Europe.

  8. 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.

  9. Cloning and characterization of prophenoloxidase A3 (proPOA3) from Culex pipiens pallens

    PubMed Central

    Zhou, Dan; Sun, Yan; Hao, Shunhua; Chen, Lin; Xiong, Chunrong; Ma, Lei; Zhang, Donghui; Hong, Shanchao; Shi, Linna; Gong, Maoqing; Zhou, Huayun; Yu, Xinyou; Shen, Bo; Zhu, Changliang

    2012-01-01

    The prophenoloxidase subunit A3 (proPOA3) gene was cloned from Culex pipiens pallens, which had an open reading frame of 2,061 bp encoding a putative 686 amino acid protein. The deduced amino acid sequence shares 98% with proPOA3 from Cx. quinquefasciatus. ProPOA3 is expressed at all developmental stages of Cx. pipiens pallens. Significant negative correlation was observed between proPOA3 expression and deltamethrin resistance in resistant Cx. pipiens pallens. Furthermore, proPOA3 expression levels were significantly lower in deltamethrin-resistant mosquitoes than in susceptible mosquitoes collected at four locations in Eastern China. However, we did not find any substantial change in proPOA3 expression in field-collected resistant Anopheles mosquitoes. Moreover, overexpressing proPOA3 in C6/36 cells led to more sensitivity to deltamethrin treatment. In laboratory and field-collected resistant Cx. pipiens pallens, a valine to isoleucine mutation (769G>A) and two synonymous mutations (1116G>C and 1116G>A) were identified in proPOA3. In addition, the mutation frequency of 769G>A and 1116G>C increased gradually, which corresponded with raised deltamethrin resistance levels. Taken together, our study provides the first evidence that proPOA3 may play a role in the regulation of deltamethrin-resistance in Cx. pipiens pallens. PMID:22561195

  10. Culex pipiens and Culex restuans mosquitoes harbor distinct microbiota dominated by few bacterial taxa.

    PubMed

    Muturi, Ephantus J; Kim, Chang-Hyun; Bara, Jeffrey; Bach, Elizabeth M; Siddappaji, Madhura H

    2016-01-13

    Mosquitoes host diverse microbial communities that influence many aspects of their biology including reproduction, digestion, and ability to transmit pathogens. Unraveling the composition, structure, and function of these microbiota can provide new opportunities for exploiting microbial function for mosquito-borne disease control. MiSeq® sequencing of 16S rRNA gene amplicons was used to characterize the microbiota of adult females of Culex pipiens L. and Cx. restuans Theobald collected from nine study sites in central Illinois. Out of 195 bacterial OTUs that were identified, 86 were shared between the two mosquito species while 16 and 93 OTUs were unique to Cx. pipiens and Cx. restuans, respectively. The composition and structure of microbial communities differed significantly between the two mosquito species with Cx. restuans hosting a more diverse bacterial community compared to Cx. pipiens. Wolbachia (OTU836919) was the dominant bacterial species in Cx. pipiens accounting for 91% of total microbiota while Sphingomonas (OTU817982) was the dominant bacterial species in Cx. restuans accounting for 31% of total microbiota. Only 3 and 6 OTUs occurred in over 60% of individuals in Cx. pipiens and Cx. restuans, respectively. There was little effect of study site on bacterial community structure of either mosquito species. These results suggest that the two mosquito species support distinct microbial communities that are sparsely distributed between individuals. These findings will allow investigations of the role of identified microbiota on the spatial and temporal heterogeneity in WNV transmission and their potential application in disease control.

  11. 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

  12. Neuroendocrine diffuse system of the respiratory tract of Rana temporaria: an immunocytochemical study.

    PubMed

    Bodegas, M E; Montuenga, L M; Sesma, P

    1995-11-01

    The neuroendocrine cell population of the respiratory system of Rana temporaria has been studied by means of immunocytochemical methods at the light-microscopic level. Isolated or clustered endocrine cells have been found in the epithelium of the buccal cavity, glottis, larynx, and lung. Nine different types of endocrine isolated cell types can be distinguished according to their immunoreactivity to several regulatory peptides [calcitonin, substance P, bombesin, peptide histidine isoleucine (PHI), cholecystokinin (CCK), and endothelin 1] and neuroendocrine markers (7B2, chromogranin, and serotonin). Neuroepithelial bodies are innervated clusters of cells simultaneously immunoreactive for serotonin and 7B2. Nerves and/or neurons have been detected in different regions of the respiratory system using antibodies against protein gene product 9.5, serotonin, calcitonin gene-related peptide (CGRP), substance P, PHI, helodermin, and CCK.

  13. 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.

  14. Maternal control of sex ratio in Rana rugosa: evidence from DNA sexing.

    PubMed

    Sakisaka, Y; Yahara, T; Miura, I; Kasuya, E

    2000-11-01

    Parental control of primary sex ratio has been reported in a mammal (red deer), some birds, and a snake. However, it remains uncertain whether other vertebrates including Amphibia can control sex ratio. In this paper, we examined the possibility in a wild population of the Japanese frog Rana rugosa which has female heterogamety. Sex ratios of their eggs were determined using DNA markers. The eggs were sampled in the field from May to August in 1998. Each egg was then sexed by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) using a sex-specific DNA marker. The result showed a male bias early in the season which changed to a female bias later, suggesting that females of R. rugosa can control the primary sex ratio.

  15. Repellent effect of Lagenaria siceraria extracts against Culex pipiens.

    PubMed

    Hassan, Mostafa I; Fouda, Mohamad A; Hammad, Kotb M; Tanani, Mohamad A; Shehata, Ahmed Z

    2014-04-01

    Ethanolic, acetone and petroleum ether extracts from leaves and stems of Lagenaria siceraria (Cucurbitaceae) were screened for their repellency effect against Culex pipiens L. mosquito. The repellent action of the present plant extracts were varied depending on the plant parts and the dose of extract. The petroleum ether extract of leaves showed the same repellency percent (100%) of commercial formulation, N. N.diethyl toulamide (DEET) at the higher dose (3.33 mg/cm2), while petroleum ether extract from stems exhibiting the repellent action (89.6%) at the same dose, respectively. Ethanolic extracts of leaves and stems exhibited the lowest repellent activity as it recorded (81.3% and 69.1%) at (6.67 mg/cm2), respectively. Results of this study may contribute to design an alternative way to control mosquitoes currently based on applications of synthetic insecticides. These extracts could be developed commercially as an effective personal protection measure against mosquito bites and thus to control diseases caused by mosquito-borne pathogens.

  16. Tracking factors modulating cytoplasmic incompatibilities in the mosquito Culex pipiens.

    PubMed

    Duron, Olivier; Bernard, Clotilde; Unal, Sandra; Berthomieu, Arnaud; Berticat, Claire; Weill, Mylène

    2006-09-01

    Wolbachia are maternally inherited endosymbiotic bacteria that infect many arthropod species and may induce cytoplasmic incompatibility (CI), resulting in abortive embryonic development. One Wolbachia host, Culex pipiens complex mosquitoes, displays high levels of variability in both CI crossing types (cytotypes) and DNA markers. We report here an analysis of 14 mosquito strains, containing 13 Wolbachia variants, and with 13 different cytotypes. Cytotypes were Wolbachia-dependent, as antibiotic treatment rendered all strains tested compatible. Cytotype distributions were independent of geographical distance between sampling sites and host subspecies, suggesting that Wolbachia does not promote a reproductive isolation depending on these parameters. Backcross analysis demonstrated a mild restoring effect of the nuclear genome, indicating that CI is mostly cytoplasmically determined for some crosses. No correlation was found between the phenotypic and genotypic variability of 16 WO prophage and transposon markers, except for the WO prophage Gp15 gene, which encodes a protein similar to a bacterial virulence factor. However, Gp15 is partially correlated with CI expression, suggesting that it could be just linked to a CI gene.

  17. 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

    2016-09-07

    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.

  18. Geographic variation in the response of Culex pipiens life history traits to temperature.

    PubMed

    Ruybal, Jordan E; Kramer, Laura D; Kilpatrick, A Marm

    2016-02-29

    Climate change is predicted to alter the transmission of many vector-borne pathogens. The quantitative impact of climate change is usually estimated by measuring the temperature-performance relationships for a single population of vectors, and then mapping this relationship across a range of temperatures or locations. However, life history traits of different populations often differ significantly. Specifically, performance across a range of temperatures is likely to vary due to local adaptation to temperature and other factors. This variation can cause spatial variation in pathogen transmission and will influence the impact of climate change on the transmission of vector-borne pathogens. We quantified variation in life history traits for four populations of Culex pipiens (Linnaeus) mosquitoes. The populations were distributed along altitudinal and latitudinal gradients in the eastern United States that spanned ~3 °C in mean summer temperature, which is similar to the magnitude of global warming expected in the next 3-5 decades. We measured larval and adult survival, development rate, and biting rate at six temperatures between 16 and 35 °C, in a common garden experiment. Temperature had strong and consistent non-linear effects on all four life history traits for all four populations. Adult female development time decreased monotonically with increasing temperature, with the largest decrease at cold temperatures. Daily juvenile and adult female survival also decreased with increasing temperature, but the largest decrease occurred at higher temperatures. There was significant among-population variation in the thermal response curves for the four life history traits across the four populations, with larval survival, adult survival, and development rate varying up to 45, 79, and 84 % among populations, respectively. However, variation was not correlated with local temperatures and thus did not support the local thermal adaptation hypothesis. These results suggest

  19. “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

  20. 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.

  1. 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

  2. An approach to mosquito control: using the dominant attraction of flowering Tamarix jordanis trees against Culex pipiens.

    PubMed

    Schlein, Yosef; Müller, Gunter C

    2008-05-01

    In this study, we identified blossoms that attract Culex pipiens L. s.l. in a Mediterranean habitat by using branches of 26 common plant species as baits for traps. The highest catch, 60.5% of the total, by flowers of Tamarix jordanis Boiss., was approximately 6 times greater than the 10.7% caught by flowering Polygonum equisetiforme Sm., and 10 times higher than the 6.6% caught by flowers of Acacia saligna (Lindle) H. L. Wendl. The catch elicited by the other plants ranged between 4.0 and 0.1%. Plant attraction also was evaluated in a field situation. Experimental and control sites were similar strips of vegetation along water channels with T. jordanis trees in the center. In the experimental site, these trees were sprayed with sucrose solution, food dye, and oral insecticide (Spinosad). Concurrently, patches of plant species and trees in the control site were sprayed with solutions of sucrose and different food dye markers. Cx. pipiens populations in both sites were monitored. The highest proportion (65.2%) of the marked mosquitoes in the control site carried the dye of flowering T. jordanis. The dye of flowering P. equisetiforme and that of A. saligna were found, respectively, in 8.1 and 3.5% of the labeled mosquitoes. The marker of reed groups (Phragmites australis [Cav.] Steudel) above the water was found in 19.4% of mosquitoes, whereas the different marker of dry land reeds was found in only 0.4% of the labeled mosquitoes. In the experimental site, after treatment, the mosquitoes decreased from approximately 255 per trap to approximately 24 mosquitoes per trap, whereas the catch in the control site reached approximately 400 mosquitoes per trap.

  3. ASYMMETRICAL EFFECTS OF INTRODUCED BULLFROGS (RANA CATESBEIANA) ON NATIVE RANID FROGS IN OREGON

    EPA Science Inventory

    Introduced American Bullfrogs (Rana catesbeiana) have become widely established in the Pacific Northwest over the last century and are throught to be an important predator of native amphibians throughout the western United States. The Northern Red-Legged Frog (Rana aurora aurora...

  4. 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.

  5. 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.

  6. Experimental alteration of the relationship between the external calcium concentration and the contractile force generated by auricular trabeculae isolated from the heart of the frog, Rana pipiens.

    PubMed

    Chapman, R A

    1971-10-01

    1. The contractile strength generated by isolated frog auricular trabeculae has been determined by perfusion with high-K Ringer over a range of [Ca](o).2. Experiments are described in which the cubic relationship between the contracture tension and [Ca](o) has been changed to a square or a linear relationship.3. These results have been interpreted by proposing that three Ca compounds, whose concentrations are proportional to [Ca](o), act co-operatively at some stage of the process leading to the generation of tension.4. The change in contractile strength, determined by regular electrically evoked twitches, has been investigated at different temperatures and the results have been explained by assuming that the concentrations of the three hypothetical activating compounds vary at different rates when [Ca](o) is altered.5. The staircase response is supposed to develop as the consequence of an increase in the concentrations of the two activating Ca compounds with the slowest time constants.6. The possible physical representations of the hypothetical activating compounds are discussed.

  7. 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...

  8. 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...

  9. 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

  10. Disentangling genetic vs. environmental causes of sex determination in the common frog, Rana temporaria.

    PubMed

    Matsuba, Chikako; Miura, Ikuo; Merilä, Juha

    2008-01-08

    Understanding of sex ratio dynamics in a given species requires understanding its sex determination system, as well as access for reliable tools for sex identification at different life stages. As in the case of many other amphibians, the common frogs (Rana temporaria) do not have well differentiated sex chromosomes, and an identification of individuals' genetic sex may be complicated by sex reversals. Here, we report results of studies shedding light on the sex determination system and sex ratio variation in this species. A microsatellite locus RtSB03 was found to be sex-linked in four geographically disparate populations, suggesting male heterogamy in common frogs. However, in three other populations examined, no or little evidence for sex-linkage was detected suggesting either ongoing/recent recombination events, and/or frequent sex-reversals. Comparison of inheritance patterns of alleles in RtSB03 and phenotypic sex within sibships revealed a mixed evidence for sex-linkage: all individuals with male phenotype carried a male specific allele in one population, whereas results were more mixed in another population. These results make sense only if we assume that the RtSB03 locus is linked to male sex determination factor in some, but not in all common frog populations, and if phenotypic sex-reversals - for which there is earlier evidence from this species - are frequently occurring.

  11. Disentangling genetic vs. environmental causes of sex determination in the common frog, Rana temporaria

    PubMed Central

    Matsuba, Chikako; Miura, Ikuo; Merilä, Juha

    2008-01-01

    Background Understanding of sex ratio dynamics in a given species requires understanding its sex determination system, as well as access for reliable tools for sex identification at different life stages. As in the case of many other amphibians, the common frogs (Rana temporaria) do not have well differentiated sex chromosomes, and an identification of individuals' genetic sex may be complicated by sex reversals. Here, we report results of studies shedding light on the sex determination system and sex ratio variation in this species. Results A microsatellite locus RtSB03 was found to be sex-linked in four geographically disparate populations, suggesting male heterogamy in common frogs. However, in three other populations examined, no or little evidence for sex-linkage was detected suggesting either ongoing/recent recombination events, and/or frequent sex-reversals. Comparison of inheritance patterns of alleles in RtSB03 and phenotypic sex within sibships revealed a mixed evidence for sex-linkage: all individuals with male phenotype carried a male specific allele in one population, whereas results were more mixed in another population. Conclusion These results make sense only if we assume that the RtSB03 locus is linked to male sex determination factor in some, but not in all common frog populations, and if phenotypic sex-reversals – for which there is earlier evidence from this species – are frequently occurring. PMID:18182101

  12. Scanning electron microscopic studies on egg surface morphology and morphometrics of Culex pipiens pipiens (Lin.) (northern house mosquito) and Culex quinquefasciatus (Say) (southern house mosquito) (Diptera: Culicidae).

    PubMed

    Adham, Fatma K; Yaman, Abeer S; Mehlhorn, Heinz

    2012-08-01

    During this study the surface morphology and morphometric of Culex pipiens pipiens (Lin.) and Culex quinquefasciatus (Say) eggs were described and compared using scanning electron microscopy for the first time. The results indicate that eggs of both species appear to be similar to great extent in surface morphology. However, morphometric provide 71.621% demarking attributes out of 44 attributes at various significant levels (P < 0.05-0.001), i.e., egg length, width and ratio of length/width, attributes of micropylar apparatus including corolla, disc, mound, tubercles size, and length of tubercular rows in micropylar region, size and density of tubercles, exochorionic pore in conical-shaped regions of eggs, and size of tubercular wheel units. Structurally, the additional presence of large tubercles strengthens the micropylar region to bear various collapsing forces in these species.

  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. Hindlimb muscle fiber types in two frogs (Rana catesbeiana and Litoria caerulea) with different locomotor behaviors: histochemical and enzymatic comparison.

    PubMed

    Crockett, Christy J; Peters, Susan E

    2008-03-01

    To test how differences in locomotor behaviors may be reflected in muscle fiber-type diversity within anurans, a comparison of hindlimb muscles between the powerful terrestrial hopper, Rana catesbeiana, and the tree frog, Litoria caerulea, was done. One postural muscle (tibialis posticus, TP) and one primary hopping muscle (plantaris longus, PL), were characterized to identify muscle fiber types using standard histochemical methods. In addition, spectophotometric analysis of activity levels of the oxidative enzyme citrate synthase (CS) and the glycolytic enzyme lactate dehydrogenase (LDH) were done in each muscle. In spite of presumed differences in behavior between the species, we found no significant differences in the proportions of the identified fiber types when the muscles were compared across species. In addition, there were no significant differences in the proportions of the different fiber types between the postural versus phasic muscles within species. Within Rana, the postural muscle (TP) had greater oxidative capacity (as measured by CS activity) than did the phasic muscle (PL). Both muscles had equivalent LDH activities. Within Litoria, PL and TP did not differ in either LDH or CS activities. Both PL and TP of Litoria had less LDH activity and greater CS activity than their homologs in Rana. Thus, in spite of the uniform populations of fiber types between muscles and species, the metabolic diversity based on enzyme activity is consistent with behavioral differences between the species. These results suggest that the range of functional diversity within fiber types may be very broad in anurans, and histochemical fiber typing alone is not a clear indicator of their metabolic or functional properties.

  15. Molecular cloning and preliminary function study of iron responsive element binding protein 1 gene from cypermethrin-resistant Culex pipiens pallens

    PubMed Central

    2011-01-01

    Background Insecticide resistance jeopardizes the control of mosquito populations and mosquito-borne disease control, which creates a major public health concern. Two-dimensional electrophoresis identified one protein segment with high sequence homology to part of Aedes aegypti iron-responsive element binding protein (IRE-BP). Method RT-PCR and RACE (rapid amplification of cDNA end) were used to clone a cDNA encoding full length IRE-BP 1. Real-time quantitative RT-PCR was used to evaluate the transcriptional level changes in the Cr-IRE strain Aedes aegypti compared to the susceptible strain of Cx. pipiens pallens. The expression profile of the gene was established in the mosquito life cycle. Methyl tritiated thymidine (3H-TdR) was used to observe the cypermethrin resistance changes in C6/36 cells containing the stably transfected IRE-BP 1 gene of Cx. pipiens pallens. Results The complete sequence of iron responsive element binding protein 1 (IRE-BP 1) has been cloned from the cypermethrin-resistant strain of Culex pipiens pallens (Cr-IRE strain). Quantitative RT-PCR analysis indicated that the IRE-BP 1 transcription level was 6.7 times higher in the Cr-IRE strain than in the susceptible strain of 4th instar larvae. The IRE-BP 1 expression was also found to be consistently higher throughout the life cycle of the Cr-IRE strain. A protein of predicted size 109.4 kDa has been detected by Western blotting in IRE-BP 1-transfected mosquito C6/36 cells. These IRE-BP 1-transfected cells also showed enhanced cypermethrin resistance compared to null-transfected or plasmid vector-transfected cells as determined by 3H-TdR incorporation. Conclusion IRE-BP 1 is expressed at higher levels in the Cr-IRE strain, and may confer some insecticide resistance in Cx. pipiens pallens. PMID:22075242

  16. Vector competence of Culex pipiens quinquefasciatus (Diptera: Culicidae) for West Nile virus isolates from Florida

    PubMed Central

    Richards, Stephanie L.; Anderson, Sheri L.; Lord, Cynthia C.

    2014-01-01

    OBJECTIVES To assess vector competence (infection, dissemination and transmission) of Culex pipiens quinquefasciatus for Florida (FL) West Nile virus (WNV) isolates. METHODS West Nile virus isolates (WN-FL-03: NY99 genotype; WN-FL-05-558, WN-FL-05-2186, WN-FL-05-510: WN02 genotype) collected from different regions of FL were used for vector competence experiments in Cx. p. quinquefasciatus from Alachua County and Indian River County in FL. Mosquitoes from both colonies were fed blood containing 7.9 ± 0.2 log10 plaque-forming units WNV/ml ± SE and incubated at 28 °C for 14 days. Vector competence, including rates of infection, dissemination, and transmission, was compared between colonies for WN-FL-03 using chi-squared. Virus titres in bodies, legs and saliva were compared using ANOVA. Daily measurements of in vitro replication of WNV isolates were evaluated in Vero cells so that a standardised virus dose for each isolate could be delivered to mosquitoes. RESULTS Infection and dissemination rates were high (≥95%) and not affected by isolate or colony (infection, P = 0.679; dissemination, P = 0.799). Transmission rates were low (≤20%), detected in one colony and affected by isolate (P = 0.008). Body and leg titres differed between isolates (body titre, P = 0.031; leg titre, P = 0.044) and colonies (body titre, P = 0.001; leg titre, P = 0.013) while saliva titre did not differ between isolates (P = 0.462). CONCLUSIONS Variation in vector competence of mosquito populations may be attributed, in part, to exposures to WNV with genetic differences leading to different rates of replication in mosquitoes. Evaluation of vector competence for different WNV isolates may help us understand vector–virus interactions and, hence, the role of vectors in complex virus transmission cycles in nature. PMID:24898274

  17. Effect of melatonin on the spermatogenesis of Rana hexadactyla (Lesson).

    PubMed

    Kasinathan, S; Gregalatchoumi, S

    1988-04-01

    It has been reported that melatonin produces either progonadal or antigonadal effects in mammals, depending on the time and mode of administration. Information on the melatonin-effect on the testis in toads indicated varied changes during the breeding and hibernating seasons. The present study in Rana hexadactyla (Lesson), a continuous breeder revealed that administration of melatonin at a dosage of 50 micrograms/frog/day either in the morning or evening for a week inhibited spermatogenesis; however, when melatonin was administered for a longer period, this inhibitory effect was lost. Moreover, treatment with melatonin both in the morning and evening had no net effect on the testes.

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

    PubMed Central

    Guillemaud, T; Pasteur, N; Rousset, F

    1997-01-01

    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

  19. 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

  20. The Ecological Aspects of Culex pipiens (Diptera: Culicidae) in Central Iran

    PubMed Central

    Dehghan, Hossein; Moosa-Kazemi, Seyed Hassan; Sadraei, Javid; Soleimani, Hassan

    2014-01-01

    Background The aim was to survey the specific factors, which cause to decrease blood feeding of mosquitoes important to succeed vector control. Methods: Larval collection was carried out from fixed and variable breeding places of Yazd County, central Iran in 2009. Autogeny-Anautogeny, Stenogamy-Eurygamy, and blood preference of Culex pipiens were studied using standard mosquito cages blood meal source for Cx. pipiens females considered as the chickens and human and fed females were kept in insectary condition (16:8 L: D, 27±3 °C and 70±10% RH). The data were analyzed using SPSS Ver. 11.5 soft ware. Results: Totally, 96 females’ mosquitoes were tested for Stenogamy versus Eurygamy and 122 for blood preference assay. In the small cages (20 × 20 × 20cm) and large cage (60 × 40 × 60cm), the ability of mating and insemination rates were 60.0 and 67.0%, respectively. In spite of Cx. pipiens fed from sucrose 5%, none of them laying eggs in 60 × 40 × 60 cages during the study. This finding indicated the Anautogeny behavior of this species. This species was found of low tendency to human blood and almost 4 fold fed on chicken. Conclusion: The occurrence of Steno-Eurygamy, Anautogeny, and Ornithophilic behaviors of Cx. Pipiens was noted. More studies need to be carried out about the bionomics of this species to gain more data about the ecophysiological and behavioral characteristics in other parts of Iran. PMID:25629063

  1. 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.

  2. The impact of weather conditions on Culex pipiens and Culex restuans (Diptera: Culicidae) abundance: a case study in Peel Region.

    PubMed

    Wang, Jiafeng; Ogden, Nick H; Zhu, Huaiping

    2011-03-01

    Mosquito populations are sensitive to long-term variations in climate and short-term variations in weather. Mosquito abundance is a key determinant of outbreaks of mosquito-borne diseases, such as West Nile virus (WNV). In this work, the short-term impact of weather conditions (temperature and precipitation) on Culex pipiens L.-Culex restuans Theobald mosquito abundance in Peel Region, Ontario, Canada, was investigated using the 2002-2009 mosquito data collected from the WNV surveillance program managed by Ontario Ministry of Health and Long-Term Care and a gamma-generalized linear model. There was a clear association between weather conditions (temperature and precipitation) and mosquito abundance, which allowed the definition of threshold criteria for temperature and precipitation conditions for mosquito population growth. A predictive statistical model for mosquito population based on weather conditions was calibrated using real weather and mosquito surveillance data, and validated using a subset of surveillance data. Results showed that WNV vector abundance on any one day could be predicted with reasonable accuracy from relationships with mean degree-days >9 degrees C over the 11 preceding days, and precipitation 35 d previously. This finding provides optimism for the development of weather-generated forecasting for WNV risk that could be used in decision support systems for interventions such as mosquito control.

  3. 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.

  4. Peptide defenses of the Cascades frog Rana cascadae: implications for the evolutionary history of frogs of the Amerana species group.

    PubMed

    Conlon, J Michael; Al-Dhaheri, Ahmed; Al-Mutawa, Eissa; Al-Kharrge, Rokaya; Ahmed, Eman; Kolodziejek, Jolanta; Nowotny, Norbert; Nielsen, Per F; Davidson, Carlos

    2007-06-01

    The Cascades frog Rana cascadae belongs to the Amerana (or Rana boylii) group that includes six additional species from western North America (R. aurora, R. boylii, R. draytonii, R. luteiventris, R. muscosa, and R. pretiosa). R. cascadae is particularly susceptible to pathogenic microorganisms in the environment and populations have declined precipitously in parts of its range so that the protection afforded by dermal antimicrobial peptides may be crucial to survival of the species. Peptidomic analysis of norepinephrine-stimulated skin secretions led to the identification of six peptides with differential cytolytic activities that were present in high abundance. Structural characterization showed that they belonged to the ranatuerin-2 (one peptide), brevinin-1 (one peptide), and temporin (four peptides) families. Ranatuerin-2CSa (GILSSFKGVAKGVAKDLAGKLLETLKCKITGC) and brevinin-1CSa (FLPILAGLAAKIVPKLFCLATKKC) showed broad spectrum antibacterial activity (MIC

  5. Host defense peptides in skin secretions of the Oregon spotted frog Rana pretiosa: implications for species resistance to chytridiomycosis.

    PubMed

    Conlon, J Michael; Mechkarska, Milena; Ahmed, Eman; Coquet, Laurent; Jouenne, Thierry; Leprince, Jérôme; Vaudry, Hubert; Hayes, Marc P; Padgett-Flohr, Gretchen

    2011-06-01

    Population declines due to chytridiomycosis among frogs belonging to the Amerana (Rana boylii) species group from western North America have been particularly severe. Norepinephrine-stimulated skin secretions from the Oregon spotted frog Rana pretiosa Baird and Girard, 1853 were collected from individuals that had been previously infected with the causative agent Batrachochytrium dendrobatidis but had proved resistant to developing chytridiomycosis. These secretions contained a more diverse array of antimicrobial peptides than found in other species from the Amerana group and 14 peptides were isolated in pure form. Determination of their primary structures identified the peptides as esculentin-2PRa and -2PRb; ranatuerin-2PRa, -2PRb, -2PRc, -2PRd, and -2PRe; brevinin-1PRa, -1PRb, -1PRc, and -1PRd; and temporin-PRa, -PRb, and -PRc. The strongly cationic ranatuerin-2PRd and the esculentin-2 peptides, which have not been identified in the secretions of other Amerana species except for the closely related R. luteiventris, showed the highest growth inhibitory potency against microorganisms. The strongly hydrophobic brevinin-1PRd was the most cytotoxic to erythrocytes. Although no clear correlation exists between production of dermal antimicrobial peptides by a species and its resistance to fatal chytridiomycosis, the diversity of these peptides in R. pretiosa may be pivotal in defending the species against environmental pathogens such as B. dendrobatidis. Copyright © 2011 Elsevier Ltd. All rights reserved.

  6. 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.

  7. "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. Copyright © 2011 Elsevier B.V. All rights reserved.

  8. 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.

  9. The forms of the Culex pipiens complex in East Asia, with ecological thoughts on their origin and interrelation.

    PubMed

    Mogi, Motoyoshi

    2012-12-01

    In East Asia, 4 forms of the Culex pipiens complex have been confirmed. A form pipiens s. s. (anautogeous pipiens) has been confirmed only in westernmost China. A temperate form pallens and a subtropical and tropical form quinquefasciatus are connected with intermediates in morphology and many aspects of ecology, but their difference is rather clear in climatic adaptation traits. The distribution of a form molestus overlaps with pallens and, in Taiwan, quinquefasciatus, and, in East Asia, this form requires artificial underground habitats for its persistence. The origin and interrelation of 3 forms other than pipiens s. s. are considered from ecological aspects, especially climatic adaptation. A hypothesis is presented that molestus was originally a form having adapted to the Mediterranean climate in the western Palaearctic, secondarily colonized artificial underground habitats, and reached East Asia in the early 20th century by ships from North America. At present, it is difficult to assign pallens with certainty to either the old or new groups of the Manchurian mosquito fauna. Three hypotheses about the interrelation between pallens and quinquefasciatus are compared, and the strengths and problems of each hypothesis are indicated. Finally, a map to show the distribution of the forms of the Culex pipiens complex before it was extensively changed by humans is presented as an initial trial. The Culex pipiens problems now troubling humans are largely human-made problems.

  10. 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.

  11. Preparation of specific antiserum against Rana esculenta pre-alpha lens crystallin.

    PubMed

    Brahma, S K; van Doorenmaalen, W J

    1976-01-01

    Specific antiserum against Rana esculenta lens pre-alpha crystallin was prepared in a rabbit by injecting antigen-antibody precipitate of this crystallin obtained from immunoelectrophoresis of esculenta total soluble lens proteins against homologous antiserum.

  12. 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.

  13. The Effect of West Nile Virus Infection on the Midgut Gene Expression of Culex pipiens quinquefasciatus Say (Diptera: Culicidae)

    PubMed Central

    Smartt, Chelsea T.; Shin, Dongyoung; Anderson, Sheri L.

    2016-01-01

    The interaction of the mosquito and the invading virus is complex and can result in physiological and gene expression alterations in the insect. The association of West Nile virus (WNV) and Culex pipiens quinquefasciatus mosquitoes results in measurable changes in gene expression; 22 gene products were shown previously to have altered expression. Sequence analysis of one product, CQ G1A1, revealed 100% amino acid identity to gram negative bacteria binding proteins (CPQGBP) in Cx. p. quinquefasciatus, Aedes aegypti (70%) and Anopheles gambiae (63%) that function in pathogen recognition. CQ G1A1 also was differentially expressed following WNV infection in two populations of Cx. p. quinquefasciatus colonized from Florida with known differences in vector competence for WNV and showed spatial and temporal gene expression differences in midgut, thorax, and carcass tissues. These data suggest gene expression of CQ G1A1 is influenced by WNV infection and the WNV infection-controlled expression differs between populations and tissues. PMID:27999244

  14. Genetic Characterization of Esterase Activity Variants Associated with an Esterase Gene Amplification in a Strain of Culex pipiens from California.

    PubMed

    Ferrari, James A

    2015-03-01

    In the Culex pipiens complex, a common mechanism of insecticide resistance is amplification of esterase genes leading to overproduction of detoxifying esterase enzymes. A number of electrophoretic esterase alleles have been identified, and in field populations individuals with the same esterase electromorph can exhibit a wide range of esterase enzyme activities. We isolated and characterized esterase activity variants associated with the esterase B1 electromorph from a field strain. A mating scheme was used to isolate chromosomes with esterase genes from the strain into 45 families. Twenty-six of the families received esterase genes from the field strain that conferred elevated esterase activity. Mean esterase activities in these families ranged from 43 to 695 nmoles α-naphthyl acetate hydrolyzed/min/mg protein. Variance components indicated that genetic variance (i.e., genetic differences among families) accounted for 77% of the total variation in esterase activity. A comparison of mean esterase activities indicated that there were at least 11 different esterase activity variants contributing to the observed genetic variation in esterase activity among the 26 families. The relevance of these results to understanding the dynamics of amplified esterase genes in populations is discussed.

  15. Polychlorinated biphenyls, sex steroid hormones and liver retinoids in adult male European common frogs Rana temporaria.

    PubMed

    Mikkelsen, Mattis; Jenssen, Bjørn Munro

    2006-05-01

    Declines in amphibian populations and species biodiversity during the last decades has called for an assessment of possible threats to these animals. Polychlorinated biphenyls (PCBs) are known endocrine disrupting contaminants and are found in high levels in some populations of wild living amphibians. To evaluate the endocrine disrupting potential of PCBs in adult frogs, Aroclor 1,254 were subcutaneously injected into male European common frogs Rana temporaria. The injected doses ranged from 0.01 to 100 mg/kg body mass, resulting in liver concentrations between 74 and 133,619 microg/kg ww. After 14 days, serum testosterone (T), estradiol (E) and hepatic retinol (R) and retinylpalmitate (RP) were easured. No dose dependent effects were found on levels of hormones or retinoids. However, a significantly higher within-group variation in the E-T ratio in the exposed groups may indicate that the sex-hormone homeostasis of male R. temporaria is affected by PCBs shortly after arousal from hibernation, but that the effects are subtle and that several different mechanisms are involved. The lack of direct effect on T, E, R and RP may be due to the timing of exposure (shortly after arousal from hibernation), or due to a relatively short exposure time to Aroclor 1,254. Based on the results, we propose that future research should focus on effects of PCBs in relation to the different physiological phases frogs experience throughout the year (hibernation, reproduction etc.).

  16. Batrachochytrium dendrobatidis infection dynamics in the Columbia spotted frog Rana luteiventris in north Idaho, USA.

    PubMed

    Russell, Danelle M; Goldberg, Caren S; Waits, Lisette P; Rosenblum, Erica Bree

    2010-11-01

    The pathogenic chytrid fungus Batrachochytrium dendrobatidis (Bd) is contributing to amphibian declines worldwide. Temperature plays an important role in both pathogen growth and host immune function, but little is known about seasonal dynamics of Bd infection in north temperate regions. Our objective was to increase understanding of Bd disease ecology by investigating patterns of Bd infection of Columbia spotted frogs Rana luteiventris across seasons, age classes, and sexes in north Idaho, USA. We collected skin swabs from 223 R. luteiventris in spring, summer, and fall 2009 at 7 ponds in the Palouse region and quantified Bd zoospores for each sample using quantitative PCR. Across seasons, Bd prevalence of adults was higher in summer than in spring or fall, suggesting that individuals may be clearing low-level infections over the summer. Among age classes, all but one late stage tadpole (Gosner stage 43-45) tested negative for Bd. Conversely, 100% of metamorphs tested positive for Bd and had the highest Bd loads of all age classes, suggesting they may be the most vulnerable age class. Adult R. luteiventris had high infection prevalence (> 60%) in all seasons, indicating that Bd infection is maintained within populations and that adults likely serve as disease reservoirs across seasons. Among adults, we also found weak evidence for females having higher infection prevalence than males. Further laboratory and field studies are needed to determine whether there are individual and population impacts from Bd on R. luteiventris and other amphibians in north Idaho.

  17. Determination of biogeographical range: an application of molecular phylogeography to the European pool frog Rana lessonae.

    PubMed Central

    Zeisset, I.; Beebee, T. J.

    2001-01-01

    Understanding how species are constrained within their biogeographical ranges is a central problem in evolutionary ecology. Essential prerequisites for addressing this question include accurate determinations of range borders and of the genetic structures of component populations. Human translocation of organisms to sites outside their natural range is one factor that increasingly complicates this issue. In areas not far beyond presumed natural range margins it can be particularly difficult to determine whether a species is native or has been introduced. The pool frog (Rana lessonae) in Britain is a specific example of this dilemma . We used variation at six polymorphic microsatellite loci for investigating the phylogeography of R. lessonae and establishing the affinities of specimens from British populations. The existence and distribution of a distinct northern clade of this species in Norway, Sweden and England infer that it is probably a long-standing native of Britain, which should therefore be included within its natural range. This conclusion was further supported by posterior probability estimates using Bayesian clustering. The phylogeographical analysis revealed unexpected patterns of genetic differentiation across the range of R. lessonae that highlighted the importance of historical colonization events in range structuring. PMID:11370966

  18. 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

    Rosà, Roberto; Marini, Giovanni; Bolzoni, Luca; Neteler, Markus; Metz, Markus; Delucchi, Luca; Chadwick, Elizabeth A; Balbo, Luca; Mosca, Andrea; Giacobini, Mario; Bertolotti, Luigi; Rizzoli, Annapaola

    2014-06-12

    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. 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. 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). Climate data collected early in the year, in conjunction with local land

  19. 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

  20. Pulmonary diffusing capacity of the bullfrog (Rana catesbeiana).

    PubMed

    Glass, M L; Burggren, W W; Johansen, K

    1981-12-01

    Carbon monoxide diffusing capacity of the lungs (DLCO) was measured in bullfrogs, Rana catesbeiana (mean body weight 260 g) along with oxygen uptake, pulmonary perfusion and lung volume. The measurements were all performed by methods depending on mass-spectrometry. Pulmonary oxygen uptake, DLCO and perfusion all increased with body temperature. At 20 degrees C O2-uptake was 0.49 ml STPD . kg-1 . min-1 at rest and DLCO was 0.022 ml STPD . kg-1 . min-1 . Torr-1. At 30 degrees C the values for O2-uptake and DLCO approximately doubled. Lung volume was large (90 ml . kg-1) and independent of body temperature. Oxygen uptake and DLCO of the bullfrog were small compared to values for a similar-sized mammal but the ratios of DLCO to O2-uptake quite similar. Analysis of available data on DLCO and O2-uptake in ectotherms also suggests a close correlation between O2-uptake and DLCO.

  1. Multiple paternity in Rana dalmatina, a monogamous territorial breeding anuran.

    PubMed

    Lodé, Thierry; Lesbarrères, David

    2004-01-01

    Polyandry and sperm competition in anurans have rarely been documented. We investigated the genetic paternity inferred from allozyme variations in 650 tadpoles from four natural ponds in a territorial breeding anuran, Rana dalmatina. Multiple paternity was demonstrated, although R. dalmatina is regarded as a monogamous species. Polyandrous mating was not a common event, occurring only in 17.9% of clutches, with no significant differences among clutches. The proportions of tadpoles fathered by a second male did not significantly differ among ponds, showing that multipaternity was not restricted to a single site. Such a polyandry may result from synchronous multiple amplexus and should reduce the heterozygote deficit related to the breeding-pond fidelity usually exhibited by most anurans.

  2. Bacterial pathogens isolated from cultured bullfrogs (Rana castesbeiana).

    PubMed

    Mauel, Michael J; Miller, Debra L; Frazier, Kendall S; Hines, Murray E

    2002-09-01

    A commercial bullfrog (Rana castesbeiana) operation in south Georgia had multiple epizootics of systemic bacterial infections over a 3-year period, 1998-2000. A number of potential pathogens (Aeromonas hydrophila, Chryseobacterium (Flavobacterium) meningosepticum, Chryseobacterium (Flavobacterium) indolgenes, Edwardsiella tarda, Citrobacterfreundii, Pseudomonas spp., and (Streptococcus iniae) were isolated from various tissues. Clinically, frogs demonstrated acute onset of torticolis, stupor, and indifference to stimuli. Cutaneous hyperemia, subcutaneous and muscular hemorrhage, and peripheral edema were consistent gross findings. Histologically, clusters of lymphocytes, monocytes, and occasional acidophiles with scattered granulomas occurred in liver, kidney, and spleen. This is the first report of S. inae and C. meningosepticum as potential disease agents in R. castesbeiana. These findings suggest that a variety of bacteria may be associated with redleg and that culture results must be obtained for accurate diagnosis.

  3. Interdigitating cells in the thymus of the frog, Rana temporaria.

    PubMed

    Bigaj, J; Płytycz, B

    1987-01-01

    Interdigitating cells (IDC) of the thymic medulla of the frog, Rana temporaria, collected in the summer, were examined by electron microscopy. The most characteristic cytological features of IDC are voluminous electron-lucent cytoplasm and widespread interdigitations and invaginations of the cell membrane. IDC possess an excentrically located nucleus with pronounced nucleoli and a thin rim of a dense chromatine as well as a perinuclear area with characteristic tubulo-vesicular complex. In our material Birbeck granules were absent. Some IDC contain phagocytized material. A few transitional forms between monocytes and IDC were observed. On the basis of these observations it is highly probable that the amphibian IDC belong to the mononuclear phagocyte system.

  4. The problem of the Culex pipiens complex in the South Pacific (including Australia)*

    PubMed Central

    Dobrotworsky, N. V.

    1967-01-01

    There are three representatives of the Culex pipiens complex in the South Pacific. C. p. fatigans is the most common and most widely distributed subspecies; it is closely associated with man. The males can be readily distinguished by the structure of the phallosome of the terminalia. C. p. molestus is spread over the southern part of Australia and in Tasmania; it also is a domestic mosquito. Throughout its extensive range in Australia, it exhibits all the biological traits that distinguish it from C. p. pipiens. C. p. australicus is widely distributed over the mainland of Australia and in Tasmania. It is superficially similar to C. p. fatigans but can be distinguished from C. p. pallens by the structure of the phallosome. It is primarily a rural non-man-biting mosquito. C. p. australicus is probably a relatively ancient member of the Australian fauna that may have evolved in the southern temperate zone. ImagesFIG. 3FIG. 4FIG. 2 PMID:5300062

  5. Cloning and characterization of 40S ribosomal protein S4 gene from Culex pipiens pallens.

    PubMed

    Hu, Xiaobang; Wang, Weijie; Zhang, Donghui; Jiao, Jianhua; Tan, Wenbin; Sun, Yan; Ma, Lei; Zhu, Changliang

    2007-02-01

    The 40S ribosomal protein S4 gene (RPS4) has been cloned from Culex pipiens pallens. An open reading frame (ORF) of 789 bp was found to encode a putative 262 amino acid protein. The deduced amino acid sequence shares 96% and 91% identity with RPS4 genes from Aedes and Anopheles respectively. Transcript expression of RPS4 was determined by real-time PCR in all life stages of deltamethrin-susceptible and -resistant strains. The results demonstrated that this gene is expressed at all developmental stages. Meanwhile, in pupae and adults, RPS4 is overexpressed in deltamethrin-resistant strain than in -susceptible strain. Our data for the first time suggests that increased expression of the RPS4 gene may play some role in the development of deltamethrin resistance in C. pipiens pallens.

  6. Decreased Flight Activity in Culex pipiens (Diptera: Culicidae) Naturally Infected With Culex flavivirus.

    PubMed

    Newman, Christina M; Anderson, Tavis K; Goldberg, Tony L

    2016-01-01

    Insect-specific flaviviruses (ISFVs) commonly infect vectors of mosquito-borne arboviruses. To investigate whether infection with an ISFV might affect mosquito flight behavior, we quantified flight behavior in Culex pipiens L. naturally infected with Culex flavivirus (CxFV). We observed a significant reduction in the scotophase (dark hours) flight activity of CxFV-positive mosquitoes relative to CxFV-negative mosquitoes, but only a marginal reduction in photophase (light hours) flight activity, and no change in the circadian pattern of flight activity. These results suggest that CxFV infection alters the flight activity of naturally infected Cx. pipiens most dramatically when these vectors are likely to be host seeking and may therefore affect the transmission of medically important arboviruses. © The Authors 2015. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  7. 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.

  8. Laboratory Evaluation of Temephos against Anopheles stephensi and Culex pipiens Larvae in Iran

    PubMed Central

    Abai, Mohammad Reza; Hanafi-Bojd, Ahmad Ali; Vatandoost, Hassan

    2016-01-01

    Background: Malaria is still a health problem in Iran. There are several vector control activities, including Indoor Residual spraying, using insecticide treated nets and larviciding including Temephos. In addition nuisance mosquitos are prevalent in the urban areas. So that evaluation of this species to larvicide will provide a clue for management of vector control activities. Methods: Two mosquito species were used in this study: Anopheles stephensi were collected from Kazeroun and Culex pipiens from Tehran, capital of Iran. All the tests were carried out according to the WHO method. All the test kis was provided by WHO. Results: Results showed a LC50= 0.0523 and LC90=0.3822 mg/l for An. stephensi. The figure for Cx. pipiens was 0.1838 and 0.8505 mg/l respectively. Conclusion: monitoring of insecticide resistance to Temephos should be evaluated regularly for management of vector control. PMID:28032103

  9. 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. © The Author 2015. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.

  10. 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

  11. Habitat use and home range of the endangered gold-spotted pond frog (Rana chosenica).

    PubMed

    Ra, Nam-Yong; Sung, Ha-Cheol; Cheong, Seokwan; Lee, Jung-Hyun; Eom, Junho; Park, Daesik

    2008-09-01

    Because of their complex life styles, amphibians and reptiles living in wetlands require both aquatic and terrestrial buffer zones in their protected conservation areas. Due to steep declines in wild populations, the gold-spotted pond frog (Rana chosenica) is listed as vulnerable by the IUCN. However, lack of data about its movements and use of habitat prevents effective conservation planning. To determine the habitat use and home range of this species, we radio-tracked 44 adult frogs for 37 days between 10 July and 4 Nov. 2007 to observe three different populations in the breeding season, non-breeding season, and late fall. The gold-spotted pond frog was very sedentary; its daily average movement was 9.8 m. Frogs stayed close to breeding ponds (within 6.6 m), and did not leave damp areas surrounding these ponds, except for dormancy migration to terrestrial sites such as dried crop fields. The average distance of dormancy migration of seven frogs from the edge of their breeding ponds was 32.0 m. The average size of an individual's home range was 713.8 m(2) (0.07 ha). The year-round population home range, which accounts for the home ranges of a population of frogs, was determined for two populations to be 8,765.0 m(2) (0.88 ha) and 3,700.9 m(2) (0.37 ha). Our results showed that to conserve this endangered species, appropriately sized wetlands and extended terrestrial buffer areas surrounding the wetlands (at least 1.33 ha, diameter 130 m) should be protected.

  12. Isolation and identification of α-proteobacteria from Culex pipiens (Diptera Culicidae) larvae.

    PubMed

    Tranchida, M C; Riccillo, P M; Rodriguero, M S; García, J J; Micieli, M V

    2012-01-01

    A survey of drainage ditches in suburban areas of La Plata, Buenos Aires province, Argentina for pathogens of Culex pipiens larvae was conducted from 2003 to 2006. C. pipiens larvae of opaque, white color were found in several of those field collections. When the white larvae were dissected and observed by phase-contrast microscopy in wet-mount preparations, the presence of bacteria, located in the hemocoel, was recorded. Laboratory experiments were performed to elucidate the pathway for transmission of this pathogen. Although approaches involving traditional culturing had failed to reveal the identity of the new microorganism present, molecular techniques to identify the pathogen in the studies reported here were successful. The partial sequence of the 16S-rRNA gene constitutes a powerful tool for the detection of new isolates from the hemocoele of C. pipiens larvae. These bacteria were characterized as belonging to the genus Novispirillum. In spite of the genus's wide distribution in different aquatic environments, information related to the parasitic relationship of Novispirillum spp. to aquatic insects is scarce, and this association has not been described in other mosquito species. This report constitutes the first documentation of Novispirillum spp. as a pathogen for mosquito larvae. Copyright © 2011 Elsevier Inc. All rights reserved.

  13. 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

  14. Efficacy of fipronil-(S)-methoprene, metaflumizone combined with amitraz, and pyriprole commercial spot-on products in preventing Culex pipiens pipiens from feeding on dogs.

    PubMed

    Bouhsira, E; Fysikopoulos, A; Franc, M

    2009-08-01

    A controlled clinical trial was carried out to assess the effectiveness of pyriprole, metaflumizone combined with amitraz, and fipronil-(S)-methoprene commercial spot-on products in preventing adult female Culex pipiens pipiens from feeding on dogs. Twenty-four beagle dogs were tested for their attractiveness to the mosquitoes and ranked accordingly to produce four groups of equivalent sensitivity to mosquitoes; six were treated with the pyriprole spot-on, six with the metaflumizone combined with amitraz spot-on, six with the fipronil-(S)-methoprene spot-on, and six were left untreated. All the dogs were challenged with 50 unfed adult female Culex in cages for one hour seven days before the treatment, and one and seven days after it. The mosquitoes were checked for blood feeding after one hour and for mortality 24 hours after they had been released into the cages. There was a significant reduction in the number of engorged mosquitoes only with the fipronil-(S)-methoprene product and there were significantly more dead mosquitoes with this product and the pyriprole product, but their effects were too small for them to be recommended for use in a dirofilariosis prevention programme.

  15. Enhanced toxicity of binary mixtures of larvicidal constituents from Asarum heterotropoides root to Culex pipiens pallens (Diptera: Culicidae).

    PubMed

    Perumalsamy, Haribalan; Kim, Jun-Ran; Kim, Soon-Il; Kwon, Hyung Wook; Ahn, Young-Joon

    2012-01-01

    The toxicity of pellitorine alone or in combination with (-)-asarinin, alpha-asarone, methyleugenol, or pentadecane (1:1, 1:2, 1:3, 2:1, and 3:1 ratios) to third instars from an insecticide-susceptible KS-CP strain and -resistant DJ-CP colony of Culex pipiens pallens Coquillett was evaluated using a direct-contact mortality bioassay. The binary mixture of pellitorine and (-)-asarinin (3:1 ratio) was significantly more toxic against KS-CP larvae (0.95 mg/liter) and DJ-CP larvae (1.07 mg/liter) than either pellitorine (2.08 mg/liter for KS-CP and 2.33 mg/liter for DJ-CP) or (-)-asarinin (11.45 and 12.61 mg/liter) alone. The toxicity of the other binary mixtures (1:1, 1:2, 1:3, and 2:1 ratios) and pellitorine did not differ significantly from each other. Based on the co-toxicity coefficient (CC) and synergistic factor (SF), the three binary mixtures (1:3, 2:1, and 3:1) operated synergistically (CC, 250-390 and SF, 1.4-2.2 for KS-CP; CC, 257-279 and SF, 1.1-2.1 for DJ-CP). The binary mixtures of pellitorine and (-)-asarinin merit further study as potential larvicides for the control of insecticide-resistant mosquito populations.

  16. Effects of combined diethylcarbamazine and albendazole treatment of bancroftian filariasis on parasite uptake and development in Culex pipiens L.

    PubMed

    Farid, Hoda A; Hammad, Ragaa E; Hassan, Marah M; Ramzy, Reda M R; El Setouhy, Maged; Weil, Gary J

    2005-07-01

    We studied effects of combined diethylcarbamazine (DEC) and albendazole (ALB) treatment on Wuchereria bancrofti microfilaria (MF) uptake and development of infective larvae (L3) in Culex pipiens. Consenting Egyptian adults with microfilaremia (MF > 300/mL) were treated with one or seven daily doses of DEC/ALB. Laboratory-reared mosquitoes were fed on subjects before and after treatment. MF uptake and infectivity (assessed by mosquito dissection) were reduced by 89.6% and 82.9%, respectively, 12 months after single-dose treatment and by 96.2% and 99.7%, respectively, after multi-dose treatment. The L3:mosquito ratio decreased by 88% to 0.082 after single-dose treatment and by 99.8% to 0.001 after multi-dose treatment. If high coverage rates can be achieved for several annual cycles, mass drug administration (MDA) with DEC/ALB has the potential to decrease transmission to unsustainable levels and eliminate filariasis in populations. Multi-dose MDA (especially in the first year) might interrupt transmission with fewer cycles than single-dose treatment.

  17. [Resident and circulating mast cells in propulsative organs of the frog Rana temporaria].

    PubMed

    Krylova, M I

    2009-01-01

    Mast cells (MCs) of the "blood" and lymph hearts of the adult frog Rana temporaria were investigated at histochemical and ultrastructural levels. Two populations of MCs were revealed in these propulsative organs: population of resident MCs and population of circulating MCs. It has been shown that the resident cardiac MCs have an oval or elongated form and are located between atrial or ventricular myocytes and under endocardial endothelium. The resident cardiac MCs are situated in connective tissue of epicardium, too. Avascular myocardium of the frog ventricle consists of a spongy network of muscle trabeculae. We revealed circulating MCs in intertrabecular spaces and clefts of the spongy myocardium and in the blood of the main central cavity. Circulating MCs are round in shape and contain a large central nucleus enriched with condensed chromatin. They resemble the lymphocytes, but show cytoplasm filled with granules. These granules ultrastructure is much like that of the granules of the cardiac resident MCs. In the lymph heart, oval and somewhat elongated resident MCs are located in the interstitial space among cross-striated muscle fibers and among smooth muscle cells of tubular (afferent and efferent) valves. Sometimes lymphocyte-like circulating MCs are revealed in the cavity of lymph heart. Circulating MCs are also present in the lymphatics located adjacent to the lymph hearts. In certain parts of the lymphatic walls MCs are in close adhesion to the mesothelial cells lining the lymphatic cavity. Our histochemical investigation revealed that both the resident and circulating MCs of the propulsative organs give a strongly positive reaction with alcian blue, but weakly red with safranin and weakly metachromatic with toluidine blue. The presence of population of circulating MCs in the frog suggests that there are differences in biology of MCs between lower and higher vertebrates.

  18. Cytonuclear discordance and historical demography of two brown frogs, Rana tagoi and R. sakuraii (Amphibia: Ranidae).

    PubMed

    Eto, Koshiro; Matsui, Masafumi

    2014-10-01

    Prior studies of mitochondrial genomic variation reveal that the Japanese brown frog Rana tagoi comprises a complex of cryptic species lineages, and that R. sakuraii arose from within this complex. Neither species forms a monophyletic group on the mitochondrial haplotype tree, precluding a simple explanation for the evolutionary origins of R. sakuraii. We present a more complete sampling of mitochondrial haplotypic variation (from the ND1 and 16S genes) plus DNA sequence variation for five nuclear loci (from the genes encoding NCX1, NFIA, POMC, SLC8A3, and TYR) to resolve the evolutionary histories of these species. We test hypotheses of population assignment (STRUCTURE) and isolation-with-migration (IM) using the more slowly evolving nuclear markers. These demographic analyses of nuclear genetic variation confirm species-level distinctness and integrity of R. sakuraii despite its apparent polyphyly on the mitochondrial haplotype tree. Divergence-time estimates from both the mitochondrial haplotypes and nuclear genomic markers suggest that R. sakuraii originated approximately one million years ago, and that incomplete sorting of mitochondrial haplotype lineages best explains non-monophyly of R. sakuraii mitochondrial haplotypes. Cytonuclear discordance elsewhere in R. tagoi reveals a case of mitochondrial introgression between two species lineages on Honshu. The earliest phylogenetic divergence within this species group occurred approximately four million years ago, followed by cladogenetic events in the Pliocene and early Pleistocene yielding 10-13 extant species lineages, including R. sakuraii as one of the youngest.

  19. 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.

  20. 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.

  1. Asymmetric competition in larval amphibian communities: conservation implications for the northern crawfish frog, Rana areolata circulosa.

    PubMed

    Parris, Matthew J; Semlitsch, Raymond D

    1998-08-01

    Asymmetric competition in larval amphibians can influence population dynamics and community structure. This density-dependent regulatory mechanism may be of particular importance for rare or endangered species such as the northern crawfish frog, Rana areolata circulosa. Interspecific competition of R. areolata with two congenerics, R. blairi and R. sphenocephala, was examined in artificial ponds. Analysis of covariance (differential mortality covariate) indicated that interspecific competition increased larval period length and decreased metamorphic body mass of R. areolata. The number of metamorphs produced was lower for R. blairi ponds when reared with R. areolata at high density. Body mass at metamorphosis was larger for R. sphenocephala when reared with R. areolata, suggesting that R. areolata facilitates larval growth in R. sphenocephala. These results indicate that the larval performance of R. areolata was reduced in the presence of interspecific competitors. Although many conservation efforts emphasize the preservation of critical habitat or particular rare species, interactive effects of biotic components in the focal community may also be important demographic regulators.

  2. 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.

  3. Behavioural response of female Culex pipiens pallens to common host plant volatiles and synthetic blends.

    PubMed

    Yu, Bao-Ting; Ding, Yan-Mei; Mo, Jian-Chu

    2015-11-17

    Most mosquito species need to obtain sugar from host plants. Little is known about the chemical cues that Culex pipiens pallens use during their orientation to nectar host plants. In this study, we investigated the behavioural responses of female Cx. pipiens pallens to common floral scent compounds and their blends. Behavioural responses of female Cx. pipiens pallens to 18 individual compounds at different concentrations were determined in the olfactometer bioassays. A synthetic blend composed of behaviourally active compounds was formulated, and its attractiveness to mosquitoes was tested. Several most attractive compounds constituted a reduced blend, and its attractiveness was tested against the solvent and the full blend, respectively. Mosquito response in the olfactometer was analyzed by comparing the percentages of mosquitoes caught in the two arms by χ(2) test (observed versus expected). Fifteen of the 18 compounds were attractive to female Cx. pipiens pallens in the dose-dependent bioassays, with the exception of β-pinene, acetophenone and nonanal. (68.00 ± 2.49) % mosquitoes responded to the full blend composed of these 15 compounds on their optimal doses when tested against the solvent, with the preference index at 46.11 ± 3.57. Six individual compounds whose preference indices were over 40 constituted the reduced blend, and it attracted (68.00 ± 1.33) % mosquitoes when tested against the solvent while its preference index was 42.00 ± 3.54. When tested against the full blend simultaneously in the olfactometer, the reduced blend could attract (45.00 ± 2.69) % of released mosquitoes, which was as attractive as the full blend. Our results demonstrate that female Cx. pipiens pallens is differentially attracted by a variety of compounds at different concentrations. Alteration of the concentration strongly affects the attractiveness of the synthetic blend. Several floral scent volatiles might be the universal olfactory cues for various

  4. 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.

  5. 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. Copyright © 2014 Elsevier Ltd. All rights reserved.

  6. An immunocytochemical and ultrastructural study of the larval anterior intestine of the frog Rana temporaria, with especial reference to endocrine cells.

    PubMed

    Bodegas, M E; Villaro, A C; Burrell, M A; Rovira, J; Valverde, E; Ortiz De Zárate, A; Sesma, P

    1997-10-01

    Endocrine cells of the larval intestine of Rana temporaria tadpoles have been identified by argyrophilic, immunocytochemical and electron-microscopical techniques. Scarce endocrine cells have been found in both the short non-absorptive zone immediately following the stomach, and in the rest of the anterior intestine. Endocrine cells are frequently seen to extend a cytoplasmic process towards the lumen. Immunoreactivity for serotonin, somatostatin, bombesin and cholecystokinin-8 has been detected. According to the ultrastructural traits of the endocrine granules, three larval intestinal endocrine populations have been differentiated.

  7. Cytoplasmic Incompatibility as a Means of Controlling Culex pipiens quinquefasciatus Mosquito in the Islands of the South-Western Indian Ocean

    PubMed Central

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

    2011-01-01

    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

  8. COMPETITION AMONG TADPOLES OF COEXISTING HEMICLONES OF HYBRIDOGENETIC RANA ESCULENTA: SUPPORT FOR THE FROZEN NICHE VARIATION MODEL.

    PubMed

    Semlitsch, Raymond D; Hotz, Hansjürg; Guex, Gaston-Denis

    1997-08-01

    Vertebrate animals reproducing without genetic recombination typically are hybrids, which have large ranges, are locally abundant, and live in disturbed or harsh habitats. This holds for the hemiclonal hybridogenetic frog Rana esculenta: it is widespread in Europe and commonly is found in disturbed habitats such as gravel pits. We hypothesize that its widespread occurrence may either be the result of natural selection for a single hemiclone acting as a broadly adapted "general-purpose" genotype, or of interclonal selection, which maintains multiple hemiclones that each are relatively narrowly adapted and perform differently across environments, that is, the Frozen Niche Variation model. We tested these competing hypotheses using 1000-L outdoor artificial ponds to rear tadpoles of the parental species (Rana lessonae [LL] and Rana ridibunda [RR]) alone, and each of three hemiclones of Rana esculenta (GUT1, GUT2, GUT3) alone, and in mixed hemiclonal populations from hatching to metamorphosis. Tadpoles of three coexisting hemiclones from a single natural population (near Gütighausen, Switzerland) were reared in both two- and three-way mixtures in equal total numbers at high and low density. For each species and hemiclone, the proportion of tadpoles metamorphosing decreased as the density of tadpoles increased, with the three hemiclones spanning the range of values exhibited by the two parental species. LL and GUT1 tadpoles produced the highest proportion of metamorphs, whereas tadpoles of RR produced the fewest metamorphs at both densities. GUT1 tadpoles also produced the largest metamorphs at low density, GUT2 and GUT3 tadpoles produced smaller metamorphs than did GUT1 tadpoles at the low density, but the three hemiclones did not differ from each other at high density. The parental species (LL and RR) were intermediate in metamorphic size to the hemiclones at low density, but all genotypes converged on a similar size at high density. Length of the larval period also

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

    USDA-ARS?s Scientific Manuscript database

    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...

  10. [Sexual receptivity of the female and insemination power of the male Culex pipiens autogenicus (Diptera, Culicidae). The cases of strict monogamy and occasional polygamy].

    PubMed

    Thanh-Xuan-Nguyen; Cabes, J L

    1977-03-14

    Females of Culex pipiens autogenicus generally have two refractory periods: post-emergence refractory period (during the first 48 hrs. after emergence) and post-insemination refractory period. Males of Culex pipiens autogenicus can inseminate 2 to 3 females. After the 4th insemination he can no longer completely fulfill his reproductory role. Therefore the female can inseminated by another male.

  11. 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.

  12. Independent degeneration of W and Y sex chromosomes in frog Rana rugosa.

    PubMed

    Miura, Ikuo; Ohtani, Hiromi; Ogata, Mitsuaki

    2012-01-01

    The frog Rana rugosa uniquely possesses two different sex-determining systems of XX/XY and ZZ/ZW, separately in the geographic populations. The sex chromosomes of both types share the same origin at chromosome 7, and the structural differences between X and Y or Z and W were evolved through two inversions. In order to ascertain the mechanisms of degeneration of W and Y chromosomes, we gynogenetically produced homozygous diploids WW and YY and examined their viability. Tadpoles from geographic group N (W(N)W(N)) containing three populations died of edema at an early developmental stage within 10 days after hatching, while tadpoles from the geographic group K (W(K)W(K)) that contained two populations died of underdeveloped growth at a much later stage, 40-50 days after fertilization. On the contrary, W(N)W(K) and W(K)W(N) hybrid embryos were viable, successfully passed the two lethal stages, and survived till the attainment of adulthood. The observed survival implies that the lethal genes of the W chromosomes are not shared by the two groups and thus demonstrates their independent degeneration histories between the local groups. In sharp contrast, a sex-linked gene of androgen receptor gene (AR) from the W chromosome was down-regulated in expression in both the groups, suggesting that inactivation of the W-AR allele preceded divergence of the two groups and appearance of the lethal genes. Besides, the YY embryos died of cardiac edema immediately after hatching. The symptom of lethality and the stage of developmental arrest differed from those for either of WW lethal embryos. We therefore conclude that the W and Y chromosomes involve no evolutionary common scenario for degeneration.

  13. Sex reversal and primary sex ratios in the common frog (Rana temporaria).

    PubMed

    Alho, Jussi S; Matsuba, Chikako; Merilä, Juha

    2010-05-01

    Sex reversal has been suggested to have profound implications for the evolution of sex chromosomes and population dynamics in ectotherms. Occasional sex reversal of genetic males has been hypothesized to prevent the evolutionary decay of nonrecombining Y chromosomes caused by the accumulation of deleterious mutations. At the same time, sex reversals can have a negative effect on population growth rate. Here, we studied phenotypic and genotypic sex in the common frog (Rana temporaria) in a subarctic environment, where strongly female-biased sex ratios have raised the possibility of frequent sex reversals. We developed two novel sex-linked microsatellite markers for the species and used them with a third, existing marker and a Bayesian modelling approach to study the occurrence of sex reversal and to determine primary sex ratios in egg clutches. Our results show that a significant proportion (0.09, 95% credible interval: 0.04-0.18) of adults that were genetically female expressed the male phenotype, but there was no evidence of sex reversal of genetic males that is required for counteracting the degeneration of Y chromosome. The primary sex ratios were mostly equal, but three clutches consisted only of genetic females and three others had a significant female bias. Reproduction of the sex-reversed genetic females appears to create all-female clutches potentially skewing the population level adult sex-ratio consistent with field observations. However, based on a simulation model, such a bias is expected to be small and transient and thus does not fully explain the observed female-bias in the field.

  14. Structural study of the frog Rana temporaria larval stomach.

    PubMed

    Rovira, J; Villaro, A C; Bodegas, M E; Valverde, E; Sesma, P

    1993-10-01

    The gastric wall of Rana temporaria tadpoles consists of a well-developed mucosa and thin muscular and serosa layers. Three cellular types--mucous, ciliated and endocrine cells--make up the lining epithelium. Different types of endocrine cells exist. Argyrophylic endocrine cells can be recognized in semithin sections of plastic-embedded material while non-argyrophylic endocrine cells can only be identified under the electron microscope. Glands are composed mainly of well-differentiated oxyntic cells and, occasionally, scarce endocrine cells. Oxyntic cells show abundant mitochondria and smooth endoplasmic reticulum, but do not contain zymogen granules as do those present in adults. Secretory canaliculi with microvilli are also well-developed. The lamina propria contains numerous vascular sinuses and nerve bundles which innervate the endothelium and some endocrine cells. The neuroendocrine regulation of frog gastric functions seems therefore to have developed in young tadpoles. Nerve fibers also innervate the muscular propria, which is composed of a single layer of smooth muscle cells. Underlying the muscle, connective fibers and a flattened layer of mesothelial cells make up the serosa. In summary, the structure of the frog larval stomach shows a well-differentiated histological pattern, especially referring to surface epithelium and glands. Some of the histological traits will also be present in adult frogs while others are characteristic of the tadpole's stage.

  15. Liver lesions produced by aflatoxins in Rana catesbeiana (bullfrog).

    PubMed

    Grassi, Tony Fernando; Pires, Paulo Wagner; Barbisan, Luis Fernando; Pai-Silva, Maeli Dal; Said, Roueda Abou; de Camargo, João Lauro Viana

    2007-09-01

    This study describes alterations induced in Rana catesbeiana (bullfrog) liver after extended dietary exposure to aflatoxins (AFs). Bullfrogs of both sexes were fed for 120 days a commercial chow blended with a rice bran-based mixture of AFs containing 667.0, 11.65, 141.74, and 3.53 mg/kg of AFs B1, B2, G1, and G2, respectively. Animals were sacrificed on study days 45, 90, and 120. Severe and progressive liver lesions with structural collapse, increased hepatocyte and biliary duct cell proliferation, appearance of basophilic hepatocytes, and diffuse scarring, were observed at all time points. There were no quantitative alterations in the liver melanomacrophage centers of the AFs-exposed animals. Increased amounts of lipid hydroperoxides, indicative of ongoing oxidative stress, were more evident in the Addutor magnum muscle than in the AFs-damaged livers. No tumors were found in the R. catesbeiana livers after 120 days of exposure to relatively high doses of AFs.

  16. Serological and bacteriological investigation of Mycobacterium ranae (fortuitum).

    PubMed

    Stanford, J L; Gunthorpe, W J

    1969-05-01

    Twenty strains of a species of fast growing Mycobacterium, some named M. fortuitum and others said to be similar to the strain of M. peregrinum, in the possession of the National Collection of Type Cultures, were examined serologically and bacteriologically. They were all found to be of the same species, but they were divisible into four serotypes (I to IV), on a basis of three variable antigens, which do not exactly correspond to the subspecies described in previous reports. The strains varied in the number of years that they had been kept in culture and were of widely divergent origin. With the exception of serotype II strains, which all came from sputa from Welsh miners, the serotypes were independent of years in culture or site of origin. Based on bacteriological and serological identity and chronological priority, we consider that M. ranae (Kuster) Bergey et al. should be reinstated in place of M. fortuitum Cruz. A modified scheme for identification of the species is proposed, which would not exclude any of the four serotypes.

  17. Ultrastructure of venom glands in the frog (Rana esculenta).

    PubMed

    De Perez, G; Hindelang, C

    1985-01-01

    Electron microscopic study of skin venom glands in the frog, Rana esculenta, revealed the syncytial structure of the inner (secretory) wall which presents two distinct zones: a basal (juxtamuscular) one, which contains nuclei and major cytoplasmic organelles, and an apical one where large electron-dense granules form and accumulate. Granules are seen to arise inside clusters of tightly packed smooth endoplasmic reticulum (SER) elements, which suggests that the SER system is mainly involved in synthesis of this material. A high glutaraldehyde concentration (5%) also reveals a poorly defined material filling the intergranular cytoplasm. No apical limits to the syncytium could be traced, which suggests massive holocrine secretion. Nerves insinuate between the muscle cells and occur all along the internal face of the muscular layer, sometimes in close contact with the syncytium. The gland duct, the wall of which consists of epidermal cells, is blocked, in contact with the gland, by an epidermal bud linked externally to the muscle layer surrounding the gland. Thus, only strong muscle tension such as to expel all or part of the epidermal bud can trigger granule release. This phenomenon can be induced by the subcutaneous injection of epinephrine, but the high and distressing dose needed to provoke appreciable changes in venom glands renders unlikely any natural intense venom release triggered by epinephrine in the frog.

  18. Seasonal cycles in testicular activity in the frog, Rana perezi.

    PubMed

    Delgado, M J; Gutiérrez, P; Alonso-Bedate, M

    1989-01-01

    Studies of seasonal testicular cycle based on spermatogenetic activity and direct measurement of plasma testosterone were made in male frog Rana perezi obtained from its natural biotope in the Iberian Peninsula. Testosterone plasma level was determined by radioimmunoassay and exhibited notable differences according to season: plasma testosterone was lowest (less than 0.5 ng/ml) in summer and then increased progressively to reach a peak in spring (3-4 ng/ml), coincident with mating. After spermiation, when an increase in temperature and photoperiod in the natural habitat occurs, levels decline. Fat bodies also show a pronounced seasonal cycle with total regression following breeding and maximal development in winter. However, testicular weight was independent of seasons, and no significant change was observed throughout the year. Histological evidence indicates that although cell nests of different types are present every month of the year, the most important spermatogenetic activity is initiated in summer. The possible relationship between spermatogenetic activity and testosterone production and the importance of environmental factors as synchronizers of seasonal reproduction are discussed.

  19. Serological and Bacteriological Investigation of Mycobacterium ranae (fortuitum)

    PubMed Central

    Stanford, J. L.; Gunthorpe, W. J.

    1969-01-01

    Twenty strains of a species of fast growing Mycobacterium, some named M. fortuitum and others said to be similar to the strain of M. peregrinum, in the possession of the National Collection of Type Cultures, were examined serologically and bacteriologically. They were all found to be of the same species, but they were divisible into four serotypes (I to IV), on a basis of three variable antigens, which do not exactly correspond to the subspecies described in previous reports. The strains varied in the number of years that they had been kept in culture and were of widely divergent origin. With the exception of serotype II strains, which all came from sputa from Welsh miners, the serotypes were independent of years in culture or site of origin. Based on bacteriological and serological identity and chronological priority, we consider that M. ranae (Kuster) Bergey et al. should be reinstated in place of M. fortuitum Cruz. A modified scheme for identification of the species is proposed, which would not exclude any of the four serotypes. Images PMID:4977473

  20. Synthesis and characterization of silver nanoparticles using Bacillus amyloliquefaciens and Bacillus subtilis to control filarial vector Culex pipiens pallens and its antimicrobial activity.

    PubMed

    Fouad, Hatem; Hongjie, Li; Yanmei, Ding; Baoting, Yu; El-Shakh, Ahmed; Abbas, Ghulam; Jianchu, Mo

    2017-11-01

    Culex pipiens pallens are the most common mosquito's vector in Asia. In order to protect the people from diseases, the anti-mosquito population is necessary that uses safe and new bio-pesticides such as bacteria-AgNPs. In our report, we used two kinds of bacteria to synthesize silver nanoparticles to examine the toxic effect on the larvae and pupae of Cx. pipiens pallens and also used as antimicrobial activity. The biosynthesis of AgNPs and its characterization was carried out by UV-Vis spectrophotometry, FTIR, TEM, SEM, and EDX. The larvicidal and pupicidal assays revealed that the lethal concentration LC50 values of Bacillus amyloliquefaciens-AgNPs were 0.72 ppm (I), 0.73 ppm (II), 0.69 ppm (III), 1.16 ppm (IV), and 4.18 (Pupae), while LC50 values of Bacillus subtilis-AgNPs were 0.60 ppm (I), 0.62 ppm (II), 0.21 ppm (III), 0.28 ppm (IV), and 3.46 ppm (Pupae) after 72 h of exposure. Antibacterial activity test of AgNPs reveals better results against rice pathogenic bacteria than bacteria alone. Thus, bacteria-mediated silver nanoparticles have a rapid effect on vector mosquito and microbial pathogen suggesting savings of energy and resources. Hence, bacteria-AgNPs may be used in the future as an effective weapon to control vector mosquito and harmful bacteria.

  1. Haematophageous vector monitoring in Djibouti city from 2008 to 2009: first records of Culex pipiens ssp. torridus (IGLISCH), and Anopheles sergentii (theobald).

    PubMed

    Faulde, Michael K; Ahmed, Ammar A

    2010-08-01

    The Horn of Africa represents a region formerly known to be highly susceptible to mosquito-borne infectious diseases. In order to monitor and analyze the current presence and threat of vector mosquitoes, continuous and standardized trapping using CDC light traps without an additional CO2-generator has been carried out at six selected monitoring sites located in Djibouti City, from August 2008 until December 2009. An overall of 620 haematophageous Diptera were trapped, 603 (97.3%) were mosquitoes, 10 (1.6%) were sand flies, and 7 (1.1%) were biting midges, respectively. Genus distribution of mosquitoes revealed that 600 (99.5%) were Culex spp., 2 (0.3%) were Anopheles sergentii, and 1 (0.2%) was Aedes aegypti. Culex species were represented by Cx. quinquefasciatus (78.5%), and Cx. pipiens ssp. torridus (21.5%). The later species was first detected focally in early December 2009 showing a strongly increasing population density resulting in a maximum trap rate of 25 mosquitoes per trap night. Sand flies were all Sergentomyia antennata, and biting midges of the genus Culicoides were represented by C. nubeculosus (71.4%) and C. vexans (28.6 %). The findings included the first records for Cx. pipiens ssp. torridus and An. sergentii in Djibouti. However, none of the captured female Culex spp, the known vector for West Nile Virus, showed positive results for viral nucleic acids using WNV RT-real time PCR system. Also, females An. sergentii were Plasmodium falciparum and P. vivax circumsporozoite protein negative.

  2. Seasonal changes in the feeding pattern of Culex pipiens pallens govern the transmission dynamics of multiple lineages of avian malaria parasites in Japanese wild bird community.

    PubMed

    Kim, K S; Tsuda, Y

    2010-12-01

    Heterogeneity in the transmission of mosquito-borne pathogens is determined largely by distribution patterns of mosquito bites among wild animal populations. Although mosquitoes are crucial for transmission of avian malaria parasites, little is known about the ecology of natural vectors. We examined bloodmeal and parasite incidence in Culex pipiens pallens by a polymerase chain reaction (PCR)-based procedure to determine how the feeding pattern of mosquitoes govern transmission dynamics of avian malaria parasites in Japanese wild birds. We collected 881 unfed and 486 blood-fed Cx. pipiens pallens resting on vegetation in a park in Tokyo. The mosquitoes were separated into abdomen and thorax prior to PCR screening. Abdomens of unfed mosquitoes were combined into 95 pools. From these, we amplified Plasmodium DNA in 32 (33.7%) pools. Among blood-fed mosquitoes, 371 individuals were screened for blood-sources and Plasmodium parasites. Plasmodium DNA was amplified from mosquitoes fed on 6 of 13 avian species identified as blood-sources. Ten Plasmodium lineages were identified on the basis of 478 bp of the cytochrome b gene, with 0.2-10% sequence divergence. The three commonest Plasmodium lineages (CXPIP09, SGS1 and PADOM02) were detected in both the abdomens and thoraxes of mosquitoes, strongly suggesting transmission of these lineages. Jungle crow (Corvus macrorhynchos) served as a natural host for the three commonest Plasmodium lineages and made up 63.8% of blood-sources. As a significant increase in feeding of vector mosquitoes on jungle crows coincided with their breeding season, jungle crows were considered to be the primary reservoir of Plasmodium transmission in this study.

  3. 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.

  4. 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.

  5. 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.

  6. Unmasking Rana okinavana Boettger, 1895 from the Ryukyus, Japan (Amphibia: Anura: Ranidae).

    PubMed

    Matsui, Masafumi

    2007-02-01

    Examination of the lectotype and a paralectotype of Rana okinavana Boettger, 1895 revealed that the species is not a brown frog of the subgenus Rana, occurring in the middle group of the Ryukyu Archipelago, but is identical with a frog of the subgenus Nidirana from the southern group of the Archipelago and Taiwan, now called R. psaltes Kuramoto, 1985. The type locality of R. okinavana given in the original description, Okinawa of the middle Ryukyus, is highly doubtful and should be somewhere in the Yaeyama Islands of the southern Ryukyus. The name R. psaltes is relegated to a subjective junior synonym of R. okinavana Boettger, 1895, while the brown frog of the subgenus Rana from the northern Ryukyus requires a replacement name.

  7. Ontogenetic changes in the epiphyseal cartilage of Rana (Pelophylax) caralitana (Anura: Ranidae).

    PubMed

    Erismis, Ugur Cengiz; Chinsamy, Anusuya

    2010-11-01

    We document histological changes through ontogeny in the epiphyseal cartilage of the third phalanx of Rana caralitana from Turkey and provide an assessment of the maturation of the epiphysis from newly metamorphosed froglets to 10-year-old individuals. The epiphysis of R. caralitana is compared to other Rana taxa previously studied, and we report on novel histological data pertaining to later stages of epiphyseal growth in this taxon. In addition, we document the development of endochondral ossification in late stages of ontogeny in R. caralitana. Our results suggest a correlation between the long lifespan of R. caralitana and the developmental changes and maturation of the epiphyseal cartilage in this taxon. This study also provides a quantitative assessment of the different regions of the epiphyseal cartilage in the epiphysis of Rana through ontogeny, and has therefore permitted quantifiable deductions about the relative maturation and differentiation of the chondrocytes of the epiphysis through time.

  8. 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.

  9. Natural vertical transmission of ndumu virus in Culex pipiens (Diptera: Culicidae) mosquitoes collected as larvae.

    PubMed

    Lutomiah, Joel; Ongus, Juliette; Linthicum, Kenneth J; Sang, Rosemary

    2014-09-01

    Ndumu virus (NDUV) is a member of the family Togaviridae and genus Alphavirus. In Kenya, the virus has been isolated from a range of mosquito species but has not been associated with human or animal morbidity. Little is know about the transmission dynamics or vertebrate reservoirs of this virus. NDUV was isolated from two pools of female Culex pipiens mosquitoes, IJR37 (n = 18) and IJR73 (n = 3), which were collected as larvae on 15 April 2013 from two dambos near the village of Marey, Ijara District, Garissa County, Kenya, and reared to adults and identified to species. These results represent the first field evidence of vertical transmission of NDUV among mosquitoes.

  10. Larvicidal activity of the extract of seaweed, Caulerpa scalpelliformis, against Culex pipiens.

    PubMed

    Cetin, Huseyin; Gokoglu, Mehmet; Oz, Emre

    2010-12-01

    The larvicidal efficacy of the acetone extract of the thalli of Caulerpa scalpelliformis var. denticulata was determined against late 2nd to early 3rd instars of Culex pipiens at concentrations ranging from 100 to 2,000 parts per million (ppm). At 1,200 ppm, the extract caused >70% larval mortality at 24-, 48-, and 72-h exposure. The LC50 (lethal concentration) and LC90 values of C. scalpelliformis were 338.91 and 1,891.31 ppm, respectively. Our data showed that this species of seaweed contains components with larvicidal properties against mosquitoes.

  11. 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.

  12. Changes in growth rate and macroelement and trace element accumulation in Hydrocharis morsus-ranae L. during the growing season in relation to environmental contamination.

    PubMed

    Polechońska, Ludmiła; Samecka-Cymerman, Aleksandra; Dambiec, Małgorzata

    2017-02-01

    The temporal variations in plant chemistry connected with its life cycle may affect the cycling of elements in an ecosystem as well as determine the usefulness of the species in phytoremediation and bioindication. In this context, there is a gap in knowledge on the role of floating plants for elements cycling in aquatic reservoirs. The aim of the study was to determine if there are variations in Hydrocharis morsus-ranae (European frog-bit) bioaccumulation capacity and the growth rate of its population during the growing season and to test the impact of environmental pollution on these features. The content of macroelements (Ca, K, Mg, N, Na, P, S) and trace metals (Cd, Co, Cu, Cr, Hg, Fe, Mn, Ni, Pb, Zn) was determined in H. morsus-ranae collected monthly from June to October from habitats differing in environmental contamination. The results showed that the highest content of most trace metals (Co, Cr, Cu, Hg, Mn, Ni, Zn) and some nutrients (N, P) in plants as well as the greatest bioaccumulation efficiency occurred simultaneously in the beginning of the growing season. In the following months, a dilution effect (manifested by a decrease in content) related to the rapid growth was observed. Co, Mn, and Ni content in plant tissues reflected the level of environmental contamination throughout the growing season which makes H. morsus-ranae a potential biomonitor of pollution for these metals. Considering the great bioaccumulation ability, high sensitivity to contamination, and low biomass of European frog-bit in polluted systems, further investigation is required to assess the real phytoremediation capability of the species.

  13. Complete mitochondrial genome of a brown frog, Rana kunyuensis (Anura: Ranidae).

    PubMed

    Li, Jiao; Yin, Wei; Xia, Rong; Lei, Guangchun; Fu, Cuizhang

    2016-01-01

    The first complete mitochondrial genome (mitogenome) of Rana sensu stricto (sensu Frost, 2013) was determined using Rana kunyuensis as a representative species. The mitogenome was 22,255 bp in length, including 13 protein-coding genes, 22 transfer RNA genes, 2 ribosomal RNA genes and duplicated control regions. The mitogenome of R. kunyuensis showed novel gene order arrangement with a translocation of tRNA(Leu)((CUN)) and ND5 in comparison with published anuran mitogenomes to date. This mitogenome should contribute to understand the evolution of anuran mitochondrial gene order arrangements.

  14. Reproductive and lipid cycles in the male frog Rana ridibunda in northern Greece.

    PubMed

    Loumbourdis, N S; Kyriakopoulou-Sklavounou, P

    1991-01-01

    1. Reproductive and lipid cycles in the male frog Rana ridibunda were studied. 2. The spermatogenesis of Rana ridibunda is of the potentially continuous type. 3. During prehibernating season (September-November) a part of lipid is mobilized from fat bodies to other body sites or is transformed to other metabolites. 4. During wintering this frog consumes mainly glycogen. 5. In February the lipid is accumulated in the fat bodies and the liver mass shows a second peak, probably as a result of glycogen accumulation. 6. The greatest decrease of metabolites was observed during the breeding season and this is the result of the intensive activities related to the reproduction and maintenance.

  15. 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.

  16. 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

  17. 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.

  18. The mosquito fauna of the western region of Spain with emphasis on ecological factors and the characterization of Culex pipiens forms.

    PubMed

    Bravo-Barriga, Daniel; Gomes, Bruno; Almeida, Antonio P G; Serrano-Aguilera, Francisco J; Pérez-Martín, Juan E; Calero-Bernal, Rafael; Reina, David; Frontera, Eva; Pinto, João

    2017-06-01

    This study updates the diversity, distribution, and seasonal trends of mosquitoes in a western region of Spain, assesses ecological determinants of Culex pipiens s.l., and determines form composition of Cx. pipiens s.s. A total of 1,495 mosquitoes of 16 species was collected during 2012-2013, of which Cx. pipiens s.l. and Cx. theileri were the most abundant. Five new records for An. maculipennis s.s., Orthopodomyia pulcripalpis, Aedes (Ochlerotatus) punctor, Cx. europaeus, and Cx. modestus were found for this region. Cx. pipiens density varied across weather and habitat patterns, correlating positively with high temperatures and with a preference for urbanized areas and rural areas within a proximity of ovine farms. Moreover, molecular identification by CQ11FL was performed in 467 Cx. pipiens s.s., detecting both pipiens (66%) and molestus (8.4%) forms coexisting in different habitats (urban, peri-urban and rural) aboveground with a high degree of hybridization (25.7%). The abundance of Cx. pipiens in urban areas and farms, with the presence of hybrids, may increase their capacity to act as bridge vectors for the transmission of arboviral infections. These data will be helpful for further implementation of entomological programs focused on risk assessment for arboviruses or other mosquito-borne pathogens. © 2017 The Society for Vector Ecology.

  19. Behavioural consistency and life history of Rana dalmatina tadpoles.

    PubMed

    Urszán, Tamás János; Török, János; Hettyey, Attila; Garamszegi, László Zsolt; Herczeg, Gábor

    2015-05-01

    The focus of evolutionary behavioural ecologists has recently turned towards understanding the causes and consequences of behavioural consistency, manifesting either as animal personality (consistency in a single behaviour) or behavioural syndrome (consistency across more behaviours). Behavioural type (mean individual behaviour) has been linked to life-history strategies, leading to the emergence of the integrated pace-of-life syndrome (POLS) theory. Using Rana dalmatina tadpoles as models, we tested if behavioural consistency and POLS could be detected during the early ontogenesis of this amphibian. We targeted two ontogenetic stages and measured activity, exploration and risk-taking in a common garden experiment, assessing both individual behavioural type and intra-individual behavioural variation. We observed that activity was consistent in all tadpoles, exploration only became consistent with advancing age and risk-taking only became consistent in tadpoles that had been tested, and thus disturbed, earlier. Only previously tested tadpoles showed trends indicative of behavioural syndromes. We found an activity-age at metamorphosis POLS in the previously untested tadpoles irrespective of age. Relative growth rate correlated positively with the intra-individual variation of activity of the previously untested older tadpoles. In previously tested older tadpoles, intra-individual variation of exploration correlated negatively and intra-individual variation of risk-taking correlated positively with relative growth rate. We provide evidence for behavioural consistency and POLS in predator- and conspecific-naive tadpoles. Intra-individual behavioural variation was also correlated to life history, suggesting its relevance for the POLS theory. The strong effect of moderate disturbance related to standard behavioural testing on later behaviour draws attention to the pitfalls embedded in repeated testing.

  20. Effects of imidacloprid on Rana catesbeiana immune and nervous system.

    PubMed

    Rios, Francesca M; Wilcoxen, Travis E; Zimmerman, Laura M

    2017-08-30

    Imidacloprid (IMD), a neonicotinoid, is generally considered to be of low toxicity in vertebrates. However, the inhibition of acetylcholine (ACh) receptors can have a profound effect on both the immune and nervous system due to the anti-inflammatory effects of ACh. Vertebrates, such as amphibians, might be affected by IMD because they breed in wetlands where the concentration of IMD is high. In our study, we experimentally exposed Rana catesbeiana tadpoles to environmentally relevant IMD and then quantified the ACh and antibody to non-replicating antigens. We hypothesized that IMD exposure would result in higher AChE and antibody levels. We completed a factorial experiment in which tadpoles were divided into four groups, two of which were exposed to 100 ng/L of IMD. After five weeks, two groups were injected with the novel antigen keyhole limpet hemocyanin (KLH) and two injected with a control. Three weeks later, tadpoles were euthanized and blood samples collected. At 100 ng/L, IMD exposure did not cause a significant difference in AChE levels or KLH-specific IgY antibodies. However, tadpoles injected with KLH had slightly higher levels of AChE. In addition, we saw a trend in total IgM with higher levels in tadpoles exposed to IMD. While we found no effect of IMD at 100 ng/L on antibody response to a novel, non-replicating antigen nor on ACh production, further research is needed to determine if higher concentrations of IMD or parasite infection can influence development of R.catesbeiana. Copyright © 2017 Elsevier Ltd. All rights reserved.

  1. 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.

  2. Fat body of the frog Rana esculenta: an ultrastructural study.

    PubMed

    Zancanaro, C; Merigo, F; Digito, M; Pelosi, G

    1996-03-01

    In the frog, the fat body is the largest body lipid deposit and is associated with the gonad. The aim of the present work was to investigate the fine structure of the fat body at different periods of the annual cycle and during prolonged starvation. Results indicate that fat body cells of Rana esculenta caught in autumn and after winter hibernation resemble mammalian adipocytes of white adipose tissue and contain markers of adipose tissue, such as S-100 protein and lipoproteinlipase. However, unlike mammalian adipocytes, fat body adipocytes consistently show small lipid droplets associated with their single, large lipid deposits, a lack of a definite external lamina, and the presence of cellular prolongations and spicula at their surfaces. Transmission and scanning electron microscopy in association with lanthanum tracer experiments suggest that in fat body adipocytes a vesicular-tubular system connects the cytoplasm and the interstitial space. In June (i.e., during the reproductive period), fat body adipocytes appear to have lost much of their lipid deposit and adjacent adipocytes show interdigitation of their plasma membranes and prominent Golgi complexes. In starved frogs, fat body cells can be almost devoid of lipid and in regression to a near-mesenchymal state. Nevertheless, these fat bodies still contain lipoproteinlipase activity (approximately 45% of that found in lipid-filled ones), indicating persistent adipose differentiation of the cells therein. Results presented here show that the R. esculenta fat body is an adipose organ undergoing reversible extreme changes in adipocyte fat content, which are associated with definite ultrastructural features. The fat body represents a suitable model for studying adipose tissue under different and extreme physiological conditions.

  3. Point Mutations Associated with Organophosphate and Carbamate Resistance in Chinese Strains of Culex pipiens quinquefasciatus (Diptera: Culicidae)

    PubMed Central

    Zhao, Minghui; Dong, Yande; Ran, Xin; Wu, Zhiming; Guo, Xiaoxia; Zhang, Yingmei; Xing, Dan; Yan, Ting; Wang, Gang; Zhu, Xiaojuan; Zhang, Hengduan; Li, Chunxiao; Zhao, Tongyan

    2014-01-01

    Acetylcholinesterase resistance has been well documented in many insects, including several mosquito species. We tested the resistance of five wild, Chinese strains of the mosquito Culex pipiens quinquefasciatus to two kinds of pesticides, dichlorvos and propoxur. An acetylcholinesterase gene (ace1) was cloned and sequenced from a pooled sample of mosquitoes from these five strains and the amino acids of five positions were found to vary (V185M, G247S, A328S, A391T, and T682A). Analysis of the correlation between mutation frequencies and resistance levels (LC50) suggests that two point mutations, G247S (r2 = 0.732, P = 0.065) and A328S (r2 = 0.891, P = 0.016), are associated with resistance to propoxur but not to dichlorvos. Although the V185M mutation was not associated with either dichlorvos or propoxur resistance, its RS genotype frequency was correlated with propoxur resistance (r2 = 0.815, P = 0.036). And the HWE test showed the A328S mutation is linked with V185M, also with G247S mutation. This suggested that these three mutations may contribute synergistically to propoxur resistance. The T682A mutation was negatively correlated with propoxur (r2 = 0.788, P = 0.045) resistance. Knowledge of these mutations may help design strategies for managing pesticide resistance in wild mosquito populations. PMID:24788312

  4. Laboratory and field trials of four repellents with Culex pipiens (Diptera: Culicidae).

    PubMed

    Coleman, R E; Richards, A L; Magnon, G J; Maxwell, C S; Debboun, M; Klein, T A; Wirtz, R A

    1994-01-01

    During Operation "Desert Shield," 16 volunteers field-tested four insect repellents (deet, the lactone CIC-4, and the piperidine compounds AI3-37220 and AI3-35765) against biting mosquitoes at King Fahd Airport, Eastern Province, Saudi Arabia. CIC-4 and AI3-37220 (25% wt/vol) provided effective (> 90%) protection against bites for 4 h. Deet and AI3-35765 protected for only 2 h. The compounds subsequently were evaluated for repellency against laboratory-reared Culex pipiens L. CIC-4 was more effective than deet, AI3-37220, or AI3-35765 at the ED50 but not at the ED95 level in initial sensitivity tests using human volunteers. At the ED95 level, deet provided significantly better protection than either piperidine compound. In laboratory duration tests, AI3-37220 provided 8 h of effective (> 90%) protection against Cx. pipiens bites, deet and AI3-35765 7 h of protection, and CIC-4 2 h of protection.

  5. 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. Copyright © 2016 Elsevier B.V. All rights reserved.

  6. 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.

  7. Amblyospora sp. (Microspora, Amblyosporidae) infecting nerve ganglia of Culex pipiens (Diptera, Culicidae) from Egypt.

    PubMed

    Darwish, A; Canning, E U

    1991-09-01

    A species of Amblyospora-infecting neurones of Culex pipiens is described. Diplokaryotic meronts, which divided by binary fission, were distinguished at the electron microscope level by their unthickened plasma membranes. Sporonts with an electron-dense surface coat gave rise to eight uninucleate sporoblasts within a sporophorous vesicle, cytoplasmic division occurring at the quadrinucleate or octonucleate stages. Indications that nuclear fusion and chromosome reorganization occurred in merogony and sporogony were obtained by light microscopy but meiosis was not detected at the ultrastructural level. Spores were typical of Amblyospora, being ovoid when fresh, truncate when stained, and having an exospore of two membranous layers subtended by a thick amorphous layer, an electron-lucent endospore, an anisofilar polar filament, and a polaroplast comprised of an anterior region of close-packed lamellae and a posterior region of expanded sacs. The metabolic products in the sporophorous vesicle took the form of large globules, small globules with electron-dense borders, and fine granules. These were depleted in mature sporophorous vesicles, though a surface layer of fine granules on the spores may have been derived from them. Many stages were degenerate and it is suggested that C. pipiens may be an accidental host in which the parasite could develop suboptimally in nervous tissue only. Infections in larvae hatched from eggs in the laboratory indicate that vertical transmission occurs.

  8. The use of morphometric wing characters to discriminate female Culex pipiens and Culex torrentium.

    PubMed

    Börstler, Jessica; Lühken, Renke; Rudolf, Martin; Steinke, Sonja; Melaun, Christian; Becker, Stefanie; Garms, Rolf; Krüger, Andreas

    2014-06-01

    The reliability of the length of wing radial vein r(2/3) as a character for the morphological discrimination of the two potential arbovirus vectors Culex pipiens s.s. and Cx. torrentium from Germany was reassessed, after this character had been neglected for more than 40 years. Additionally, multivariate morphometric analyses were applied to evaluate wing shape variation between both species. Although high-throughput molecular tools are now available to differentiate the two species, a simple, low-cost routine alternative may be useful in the absence of a molecular laboratory, such as under semi-field conditions. A thin-plate splines transformation confirmed that primarily the shrinkage of vein r(2/3) is responsible for the wing differences between the two species. In the bivariate analysis, the r(2/3)/r3 indices of Cx. pipiens s.s. and Cx. torrentium were 0.185 and 0.289, respectively, resulting in a correct classification of more than 91% of all tested specimens. Using the absolute length of vein r(2/3) alone still allowed for more than 90% accurate discrimination. Furthermore, classification accuracy of linear discriminant analysis exceeded 97%.

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

    PubMed

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

    2015-08-20

    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.

  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. Elevated couch potato transcripts associated with adult diapause in the mosquito Culex pipiens.

    PubMed

    Zhang, Qirui; Denlinger, David L

    2011-05-01

    The couch potato (cpo) cDNA that we cloned from the northern house mosquito, Culex pipiens, encodes the C-terminus containing a highly conserved RNA recognition motif (RRM). Protein structure prediction indicates a canonical RRM structure with a βαββαβ topological structure. Northern blots indicate a single mRNA band over 9.49 kb, and Southern blot analysis suggests that the cpo gene contains large introns. Highest expression was noted in first instar larvae and pupae. A comparison of nondiapausing (long daylength) and diapausing (short daylength) adult females showed no difference immediately at adult eclosion, but by day 7 and thereafter, expression of cpo was much higher in diapausing adults. When 2-month old diapausing females were transferred from short daylength to diapausing-terminating conditions of long daylength and high temperature, expression of cpo declined. Similarly, when a topical application of JH III was used to terminate diapause abundance of the cpo transcript declined. Consistent with observations in Drosophila melanogaster and several other species levels of cpo in C. pipiens are influenced by the diapause program, although the direction of change is not the same in all species.

  12. Sugar feeding improves survival of nondiapausing cold-stored Culex pipiens.

    PubMed

    Rinehart, Joseph P; Yocum, George D; Robich, Rebecca M

    2012-11-01

    The continuous culture of mosquitoes is a costly endeavor for vector biology laboratories. In addition to the resources that must be committed to colony maintenance, biological costs, including genetic drift and accidental colony loss, also can occur. Although alternatives do exist, their application to mosquitoes is limited. Mosquito cryopreservation remains elusive, and many important species lack a well-defined diapause. Previously, we demonstrated that cold storing nondiapausing mated adult females of the northern house mosquito, Culex pipiens L. resulted in a nearly four-fold increase in longevity when measured at the LT50, allowing for cold storage for up to 10 wk. In the current study, we used sugar feeding during cold storage to significantly improve cold storage longevity. At 6 degrees C, the LT50 of cold stored females was 23 wk, and 100% mortality was not realized until 43 wk. Cold-stored females did exhibit reduced fecundity, but egg production returned to normal levels within two generations. These results suggest that cold storage without diapause induction is a viable option for Cx. pipiens, and with the addition of sugar feeding, a colony could be maintained with less than two generations per year.

  13. Pathogenesis of Frog Virus 3 ( Ranavirus, Iridoviridae) Infection in Wood Frogs ( Rana sylvatica).

    PubMed

    Forzán, M J; Jones, K M; Ariel, E; Whittington, R J; Wood, J; Markham, R J Frederick; Daoust, P-Y

    2017-01-01

    Wood frogs ( Rana sylvatica) are highly susceptible to infection with Frog virus 3 (FV3, Ranavirus, Iridoviridae), a cause of mass mortality in wild populations. To elucidate the pathogenesis of FV3 infection in wood frogs, 40 wild-caught adults were acclimated to captivity, inoculated orally with a fatal dose of 10(4.43) pfu/frog, and euthanized at 0.25, 0.5, 1, 2, 4, 9, and 14 days postinfection (dpi). Mild lesions occurred sporadically in the skin (petechiae) and bone marrow (necrosis) during the first 2 dpi. Severe lesions occurred 1 to 2 weeks postinfection and consisted of necrosis of medullary and extramedullary hematopoietic tissue, lymphoid tissue in spleen and throughout the body, and epithelium of skin, mucosae, and renal tubules. Viral DNA was first detected (polymerase chain reaction) in liver at 4 dpi; by dpi 9 and 14, all viscera tested (liver, kidney, and spleen), skin, and feces were positive. Immunohistochemistry (IHC) first detected viral antigen in small areas devoid of histologic lesions in the oral mucosa, lung, and colon at 4 dpi; by 9 and 14 dpi, IHC labeling of viral antigen associated with necrosis was found in multiple tissues. Based on IHC staining intensity and lesion severity, the skin, oral, and gastrointestinal epithelium and renal tubular epithelium were important sites of viral replication and shedding, suggesting that direct contact (skin) and fecal-oral contamination are effective routes of transmission and that skin tissue, oral, and cloacal swabs may be appropriate antemortem diagnostic samples in late stages of disease (>1 week postinfection) but poor samples to detect infection in clinically healthy frogs.

  14. 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

    From 2001 to 2005, we studied the demography and seasonal movement of Oregon spotted frogs (Rana pretiosa) translocated into created ponds in Dilman Meadow in central Oregon. Our objectives were to inform future monitoring and management at the site, and to elucidate poorly known aspects of the species’ population ecology. Movement rates revealed complementary use of sites seasonally, with one small spring being preferred during winter that was rarely used during the rest of the year. Growth rates were significantly higher in ponds that were not used for breeding, and larger size resulted in significantly higher survival. When variation in survival by size was accounted for there was little variation among ponds in survival. Seasonal estimates of survival were lowest for males during the breeding/post-breeding redistribution period, suggesting a high cost of breeding for males. Overwintering survival for both genders was relatively high. Our study supports others in suggesting Oregon spotted frogs are specific in their overwintering habitat requirements, and that predator-free springs may be of particular value. We suggest that any future monitoring include measures of the rate of pond succession. Demographic monitoring should include metrics of both frog reproduction and survival: counts of egg masses at all ponds during spring, and capture-recapture study of survival in mid and late summer when capture rates are highest. Additional study of early life stages would be particularly useful to broaden our understanding of the species’ ecology. Specifically, adding intensive capture and marking effort after larval transformation in fall would enable a full understanding of the annual life cycle. Complete study of the annual life cycle is needed to isolate the life stages and mechanisms through which Oregon spotted frogs are affected by stressors such as nonnative predators. Dilman Meadow, which lacks many hypothesized stressors, is an important reference for

  15. Cadmium pollution and amphibians--Studies in tadpoles of Rana limnocharis.

    PubMed

    Patar, Arabinda; Giri, Anirudha; Boro, Freeman; Bhuyan, Krishna; Singha, Utsab; Giri, Sarbani

    2016-02-01

    Cadmium is released into the environment in increasing amounts from different natural and anthropogenic activities contaminating the aquatic habitats. Amphibian tadpoles develop in water and hence are likely to be adversely affected by cadmium present in the aquatic environment. We have studied the toxic and genotoxic effects of CdCl2 on the tadpoles of Rana limnocharis. CdCl2 in the concentration range between 0.1 and 0.4 mg/L induced significant mortality in R. limnocharis tadpoles in a dose and time dependent manner. The 10-day LC50 which has more ecological relevance was far less than the 24-h LC50. Tadpoles exposed to CdCl2 metamorphosed at an early age possibly as a survival strategy to move out of the stressful environment. The body weight of the CdCl2 exposed animals at metamorphosis was lower compared to the control individuals which may affect survival and reproductive fitness in adult life. Besides, the average body length of the metamorphosed individuals in the CdCl2 exposed group was higher than the control group. CdCl2 was found to be genotoxic in micronucleus test and comet assay. The ambient concentration of Cd could reach up to 60 μg/L or more. Exposure to 18.5 μg/L of CdCl2 (1% of 24-h LC50) induced significant increase in DNA strand breaks as compared to the control. The present findings demonstrate that presence of cadmium in the aquatic environment can significantly alter the life history traits and cause DNA damage in amphibians and hence, could contribute towards their population decline.

  16. Triiodothyronine (T3) action on aquatic locomotor behavior during metamorphosis of the bullfrog Rana catesbeiana

    PubMed Central

    Fernández-Mongil, Marisabel; Venza, Celia J.; Rivera, Amelia; Lasalde-Dominicci, José A.; Burggren, Warren; Rojas, Legier V.

    2010-01-01

    Thyroid hormones - particularly triiodothyronine, T3 - play a critical role in the morphological transformations comprising metamorphosis in larval bullfrogs (Rana catesbeiana). Traditional staging criteria for anuran larvae incompletely distinguish physiological and behavioral changes during growth. We therefore first developed a new parameter to describe larval growth, the developmental index (DI), which is simply the ratio between the tail length of the larva and its head diameter. Using the DI we were able to identify two distinct populations classifying the larvae during growth along a continuous linear scale with a cutoff value of DI at 2.8. Classification based on the DI, used in this study, proved an effective complement to existing classifications based on developmental staging into pre- or pro-metamorphic stages. Exposure to T3 in the water induced a rapid (beginning within 5 min) and significant decrease (~20–40%) in locomotor activity, measured as total distance traversed and velocity. The largest decrease occurred in more developed larvae (DI< 2.8). To determine correlated changes in the neuromuscular junctions during metamorphosis and apoptotic tail loss, miniature endplate currents from tail muscle were recorded during acute exposure to a hypertonic solution, which simulates an apoptotic volume decrease. Our results support a role for T3 in regulating larval locomotor activity during development, and suggest an enhanced response to volume depletion at the neuromuscular junction of older larvae (DI<2.8) compared to younger animals (DI≥2.8). We discuss the significance of the possible role of an apoptotic volume decrease at the level of the neuromuscular junction. PMID:19123131

  17. Enzymatic regulation of seasonal glycogen cycling in the freeze-tolerant wood frog, Rana sylvatica.

    PubMed

    do Amaral, M Clara F; Lee, Richard E; Costanzo, Jon P

    2016-12-01

    Liver glycogen is an important energy store in vertebrates, and in the freeze-tolerant wood frog, Rana sylvatica, this carbohydrate also serves as a major source of the cryoprotectant glucose. We investigated how variation in the levels of the catalytic subunit of protein kinase A (PKAc), glycogen phosphorylase (GP), and glycogen synthase (GS) relates to seasonal glycogen cycling in a temperate (Ohioan) and subarctic (Alaskan) populations of this species. In spring, Ohioan frogs had reduced potential for glycogen synthesis, as evidenced by low GS activity and high PKAc protein levels. In addition, glycogen levels in spring were the lowest of four seasonal samples, as energy input was likely directed towards metabolism and somatic growth during this period. Near-maximal glycogen levels were reached by mid-summer, and remained unchanged in fall and winter, suggesting that glycogenesis was curtailed during this period. Ohioan frogs had a high potential for glycogenolysis and glycogenesis in winter, as evidenced by large glycogen reserves, high levels of GP and GS proteins, and high GS activity, which likely allows for rapid mobilization of cryoprotectant during freezing and replenishing of glycogen reserves during thawing. Alaskan frogs also achieved a near-maximal liver glycogen concentration by summer and displayed high glycogenic and glycogenolytic potential in winter, but, unlike Ohioan frogs, started replenishing their energy reserves early in spring. We conclude that variation in levels of both glycogenolytic and glycogenic enzymes likely happens in response to seasonal changes in energetic strategies and demands, with winter survival being a key component to understanding the regulation of glycogen cycling in this species.

  18. Ranavirus in wood frogs (Rana sylvatica): potential sources of transmission within and between ponds.

    PubMed

    Harp, Elizabeth M; Petranka, James W

    2006-04-01

    Members of the genus Ranavirus (family Iridoviridae) can cause catastrophic mortality of pond-breeding amphibians and are associated with an emerging infectious disease that may be contributing to amphibian declines. We conducted three experiments to examine factors that may affect transmission both within and between local breeding populations of the wood frog (Rana sylvatica). In a laboratory study, when exposed to moribund tadpoles collected during a local ranaviral die-off, uninfected tadpoles died as soon as 4 days after exposure. The onset of death was accelerated when tadpoles were allowed to scavenge on carcasses of infected tadpoles. In a mesocosm experiment that was conducted in outdoor wading pools, die-offs of tadpoles began approximately 19 days after infected tadpoles were added to pools containing uninfected tadpoles. Mass die-offs with greater than 98% mortality occurred in all pools, regardless of the initial tadpole density. In a second mesocosm experiment, the addition of water and bottom sediments that were collected from a pond during a ranaviral die-off did not result in lower tadpole survival or growth relative to controls. Only a small percentage of tadpoles appeared to be sick, and most tadpoles survived until the first individuals began metamorphosing within a pool. However, tests for ranavirus using polymerase chain reaction were positive for most pools that received contaminated sediment, suggesting that some infections were sublethal. Our results indicate that transmission within ponds is enhanced by scavenging and that spread between local ponds could occur via the transport of contaminated sediment by animals or humans.

  19. 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.

  20. 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.

  1. [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.

  2. Hybridization and mitochondrial genome introgression between Rana chensinensis and R. kukunoris.

    PubMed

    Qi, Yin; Lu, Bin; Gao, Haiyan; Hu, Ping; Fu, Jinzhong

    2014-11-01

    Mitochondrial genome (mito-genome) introgression among metazoans is commonplace, and several biological processes may promote such introgression. We examined two proposed processes for the mito-genome introgression between Rana chensinensis and R. kukunoris: natural hybridization and sex-biased dispersal. We sampled 477 individuals from 28 sites in the potential hybrid zone in the western Tsinling Mountains. Mitochondrial gene (cyt-b) trees were used to examine the introgression events. Microsatellite DNA loci, cyt-b and morphological data were used to identify hybrids and to examine the extent of natural hybridization. We detected rampant bidirectional introgressions, both ancient and recent, between the two species. Furthermore, we found a wide hybrid zone, and frequent and asymmetric hybridization. The hybrid zone cline analysis revealed a clear mitochondrial-nuclear discordance; while most nuclear markers displayed similar and steep clines, cyt-b had a displaced cline centre and a more gradual and wider cline. We also detected strong and asymmetric historical maternal gene flow across the hybrid zone. This widespread hybridization and detected low mito-nuclear conflicts may, at least partially, explain the high frequency of introgression. Lastly, microsatellite data and population genetic methods were used to assess sex-biased dispersal. A weak pattern of female-biased dispersal was detected in both species, suggesting it may not play an important role in the observed introgression. Our data are consistent with the hybridization hypothesis, but support for the sex-biased dispersal hypothesis is weak. We further suggest that selective advantages of the R. kukunoris-type mito-genome in thermal adaptation may also contribute to the introgression between the two species. © 2014 John Wiley & Sons Ltd.

  3. 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.

  4. 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. Copyright © 2015 Elsevier Inc. All rights reserved.

  5. Recent amplification of miniature inverted-repeat transposable elements in the vector mosquito Culex pipiens: characterization of the Mimo family.

    PubMed

    Feschotte, C; Mouchès, C

    2000-05-30

    We describe a new family of repetitive elements, named Mimo, from the mosquito Culex pipiens. Structural characteristics of these elements fit well with those of miniature inverted-repeat transposable elements (MITEs), which are ubiquitous and highly abundant in plant genomes. The occurrence of Mimo in C. pipiens provides new evidence that MITEs are not restricted to plant genomes, but may be widespread in arthropods as well. The copy number of Mimo elements in C. pipiens ( approximately 1000 copies in a 540Mb genome) supports the hypothesis that there is a positive correlation between genome size and the magnitude of MITE proliferation. In contrast to most MITE families described so far, members of the Mimo family share a high sequence conservation, which may reflect a recent amplification history in this species. In addition, we found that Mimo elements are a frequent nest for other MITE-like elements, suggesting that multiple and successive MITE transposition events have occurred very recently in the C. pipiens genome. Despite evidence for recent mobility of these MITEs, no element has been found to encode a protein; therefore, we do not know how they have transposed and have spread in the genome. However, some sequence similarities in terminal inverted-repeats suggest a possible filiation of some of these mosquito MITEs with pogo-like DNA transposons.

  6. Diapause-specific gene expression in the northern house mosquito, Culex pipiens L., identified by suppressive subtractive hybridization

    USDA-ARS?s Scientific Manuscript database

    In this study we probe the molecular events underpinning diapause observed in overwintering females of Culex pipiens. Using suppressive subtractive hybridization (SSH) we have identified 40 genes that are either upregulated or downregulated during this seasonal period of dormancy. Northern blot hybr...

  7. Insecticidal and repellent activities of insecticide-sucrose solutions to Culex pipiens molestus (Diptera: Culicidae) under laboratory and field conditions.

    PubMed

    Shin, Ehyun; Park, Chan; Ahn, Young-Joon; Lee, Dong-Kyu; Chang, Kyu-Sik

    2011-06-01

    Culex pipiens molestus Forskal has been reported as a dominant species in underground structures of urban areas in the Republic of Korea (ROK) during all seasons and becomes bothersome to humans in late autumn and winter. Most Cx. pipiens molestus in septic tanks are controlled in the ROK using larvicides such as Bt and IGR. However, there are a number of problems associated with larvicides, such as high cost and requirement for frequent use. In the present work, a new control method for Cx. pipiens molestus in septic tanks by using mixtures of sucrose solution with insecticides was investigated. The insecticidal and repellent activities of ten insecticides were evaluated for best control of Cx. pipiens molestus in septic tanks. Firstly, differences in susceptibilities to insecticides were evaluated in topical assays by forced direct contact bioassay and in a screened wire cage by free direct contact bioassay. The difference in insecticide susceptibility in the mosquitoes was the result of repellency by the insecticides. In three septic tanks, the density of Culex mosquitoes was sharply reduced by a deltamethrin-sucrose solution kit. The results demonstrated the potential for mosquito control by deltamethrin-sucrose solution, and the study offers basic information related to mosquito control in septic tanks. Copyright © 2011 Society of Chemical Industry.

  8. Distribution of the members of the Pipiens Assemblage in the sympatric area from Argentina: which is where and when?

    PubMed

    Cardo, María V; Rubio, Alejandra; Junges, Melania; Vezzani, Darío; Carbajo, Aníbal E

    2016-11-01

    Given their medical and veterinary relevance, the members of the Pipiens Assemblage are a worldwide target of ecological research. The distribution of Culex pipiens s.s. and Cx. quinquefasciatus converge in Buenos Aires, Argentina, where hybrids have been detected. Each member of the assemblage exhibits a distinct eco-physiological behaviour that can affect its efficiency in pathogen transmission. Our aim was to identify the environmental drivers for the spatio-temporal distribution of each member, focusing on latitudinal and urbanisation gradients. Immatures of mosquitoes were surveyed in artificial containers found within 11 public cemeteries, raised up to the adult stage and identified by their male genitalia. The distribution of each member was associated with the environment in a Generalized Linear Model. The variable accounting for most of the heterogeneity was latitude; Cx. quinquefasciatus was collected more frequently at northern cemeteries, whereas Cx. pipiens and hybrids were more likely at the southern extreme. The urbanisation gradient was also associated with the occurrence of Cx. quinquefasciatus and hybrids at the high and low end, respectively. Other relevant variables were cemetery total area, the proportion with graves and the presence of plastic flowers in the containers. The spatial distribution of the members of the Pipiens Assemblage within the sympatric region in South America is driven by environmental features. The information presented herein provides essential baseline data for surveillance programs and control activities.

  9. Rift Valley fever virus and European mosquitoes: vector competence of Culex pipiens and Stegomyia albopicta (= Aedes albopictus).

    PubMed

    Brustolin, M; Talavera, S; Nuñez, A; Santamaría, C; Rivas, R; Pujol, N; Valle, M; Verdún, M; Brun, A; Pagès, N; Busquets, N

    2017-08-07

    Rift Valley fever (RVF) is a mosquito-borne disease caused by the Rift Valley fever virus (RVFV). Rift Valley fever affects a large number of species, including human, and has severe impact on public health and the economy, especially in African countries. The present study examined the vector competence of three different European mosquito species, Culex pipiens (Linnaeus, 1758) form molestus (Diptera: Culicidae), Culex pipiens hybrid form and Stegomyia albopicta (= Aedes albopictus) (Skuse, 1894) (Diptera: Culicidae). Mosquitoes were artificially fed with blood containing RVFV. Infection, disseminated infection and transmission efficiency were evaluated. This is the first study to assess the transmission efficiency of European mosquito species using a virulent RVFV strain. The virus disseminated in Cx. pipiens hybrid form and in S. albopicta. Moreover, infectious viral particles were isolated from saliva of both species, showing their RVFV transmission capacity. The presence of competent Cx. pipiens and S. albopicta in Spain indicates that an autochthonous outbreak of RVF may occur if the virus is introduced. These findings provide information that will help health authorities to set up efficient entomological surveillance and RVFV vector control programmes. © 2017 The Authors. Medical and Veterinary Entomology published by John Wiley & Sons Ltd on behalf of Royal Entomological Society.

  10. Distribution of the members of the Pipiens Assemblage in the sympatric area from Argentina: which is where and when?

    PubMed Central

    Cardo, María V; Rubio, Alejandra; Junges, Melania; Vezzani, Darío; Carbajo, Aníbal E

    2016-01-01

    Given their medical and veterinary relevance, the members of the Pipiens Assemblage are a worldwide target of ecological research. The distribution of Culex pipiens s.s. and Cx. quinquefasciatus converge in Buenos Aires, Argentina, where hybrids have been detected. Each member of the assemblage exhibits a distinct eco-physiological behaviour that can affect its efficiency in pathogen transmission. Our aim was to identify the environmental drivers for the spatio-temporal distribution of each member, focusing on latitudinal and urbanisation gradients. Immatures of mosquitoes were surveyed in artificial containers found within 11 public cemeteries, raised up to the adult stage and identified by their male genitalia. The distribution of each member was associated with the environment in a Generalized Linear Model. The variable accounting for most of the heterogeneity was latitude; Cx. quinquefasciatus was collected more frequently at northern cemeteries, whereas Cx. pipiens and hybrids were more likely at the southern extreme. The urbanisation gradient was also associated with the occurrence of Cx. quinquefasciatus and hybrids at the high and low end, respectively. Other relevant variables were cemetery total area, the proportion with graves and the presence of plastic flowers in the containers. The spatial distribution of the members of the Pipiens Assemblage within the sympatric region in South America is driven by environmental features. The information presented herein provides essential baseline data for surveillance programs and control activities. PMID:27783720

  11. 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...

  12. 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...

  13. 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

  14. Effects of the Chytrid fungus on the Tarahumara frog (Rana tarahumarae) in Arizona and Sonora, Mexico

    Treesearch

    Stephen F. Hale; Philip C. Rosen; James L. Jarchow; Gregory A. Bradley

    2005-01-01

    We conducted histological analyses on museum specimens collected 1975-1999 from 10 sites in Arizona and Sonora to test for the pathogenic chytrid fungus (Batrachochytrium dendrobatidis) in ranid frogs, focusing on the Tarahumara frog (Rana tarahumarae). During 1981-2000, frogs displaying disease signs were found in the field, and...

  15. Responses of foothill yellow-legged frog (Rana boylii) larvae to an introduced predator

    Treesearch

    David J. Paoletti; Deanna H. Olson; Andrew R. Blaustein

    2011-01-01

    The consequences of species introductions into non-native habitats are a major cause for concern in the U.S. Of particular interest are the effects of predation by introduced fishes on native amphibian communities. We sought to determine whether Foothill Yellow-legged Frog (Rana boylii) larvae could recognize non-native Small mouth Bass (...

  16. Influence of anuran prey on the condition and distribution of Rana muscosa in the Sierra Nevada

    Treesearch

    K.L. Pope; K.R. Matthews

    2002-01-01

    Mountain yellow-legged frogs (Rana muscosa) at high elevations of the Sierra Nevada must obtain enough food during summer to survive 7–9 winter months when their aquatic habitats are frozen and food is presumably unavailable. Adults of R. muscosa prey on a variety of organisms, including aquatic and terrestrial invertebrates and...

  17. Foothill yellow-legged frog (Rana boylii) oviposition site choice at multiple spatial scales

    Treesearch

    Amy J. Lind; Hartwell H. Welsh; Clara A. Wheeler

    2016-01-01

    Studies of resource selection at multiple scales are critical to understanding ecological and evolutionary attributes of a species. We analyzed relative abundance, habitat use, and oviposition site selection of Foothill Yellow-Legged Frogs (Rana boylii) at 11 localities across two geographic regions in California (northern Coast Range and Sierra...

  18. Site fidelity of the declining amphibian Rana sierrae (Sierra Nevada yellow-legged frog)

    Treesearch

    Kathleen Matthews; Haiganoush Preisler

    2010-01-01

    From 1997 to 2006, we used mark–recapture models to estimate the site fidelity of 1250 Sierra Nevada yellow-legged frogs (Rana sierrae) in Kings Canyon National Park, California, USA, during their three main activity periods of overwintering, breeding, and feeding. To quantify site fidelity, the tendency to return to and reuse previously occupied...

  19. Expression of leptin receptor in the oviduct of Chinese brown frog (Rana dybowskii).

    PubMed

    Xi, Liqin; Liu, Yuning; Tang, Zeqi; Sheng, Xia; Zhang, Haolin; Weng, Qiang; Xu, Meiyu

    2017-06-01

    The oviduct of Chinese brown frog (Rana dybowskii) expands specifically during prehibernation instead of in the breeding period. In this study, we investigated the expression of leptin receptor (Ob-Rb) in Rana dybowskii oviduct during the breeding period and prehibernation. Histologically, the oviduct of Rana dybowskii consists of glandular cells, tubule lumen, and epithelial cells. The oviductal weight and pipe diameter also revealed significant differences, which were higher in prehibernation than that of the breeding period. Ob-Rb was observed in stromal cells of oviductal tissue in both the breeding period and prehibernation. The mean protein and mRNA levels of the Ob-Rb were significantly higher in prehibernation as compared with the breeding period. In addition, oviductal content of leptin was also higher in prehibernation than that of the breeding period. These results suggested that oviduct of Rana dybowskii might be a target organ of leptin, and leptin may play an autocrine/paracrine role mediated by Ob-Rb in regulating the oviductal hypertrophy during prehibernation. Copyright © 2017 the American Physiological Society.

  20. Mating strategy and breeding patterns of the foothill yellow-legged frog (Rana boylii)

    Treesearch

    Clara A. Wheeler; Hartwell H. Welsh Jr.

    2008-01-01

    The Foothill Yellow-legged Frog (Rana boylii) has declined across much of its native range in California. Improper stream management may lower egg mass survival and reduce the availability of suitable breeding habitats. We collected data during six breeding-seasons (2002-2007) along an unregulated stream in northwestern California. We monitored...

  1. Oregon spotted frog (Rana pretiosa) monitoring at Jack Creek 2015-2016

    USGS Publications Warehouse

    Adams, Michael J.; Pearl, Christopher A.; Mccreary, Brome; Galvan, Stephanie; Rowe, Jennifer

    2017-01-01

    This dataset contains information from mark-recapture and egg mass surveys conducted 2015-2016 by USGS as part of an ongoing Oregon spotted frog (Rana pretiosa) monitoring effort at Jack Creek, Klamath County, Oregon. Data consist of spotted frog counts aggregated by date, location, life stage, and sex, as well as data on environmental conditions at the time each survey.

  2. 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.

  3. Oviposition preferences of Culex restuans and Culex pipiens (Diptera: Culicidae) for selected infusions in oviposition traps and gravid traps.

    PubMed

    Jackson, Bryan T; Paulson, Sally L; Youngman, Roger R; Scheffel, Sabra L; Hawkins, Belinda

    2005-12-01

    Field studies were conducted in southwestern Virginia to determine the ovipositional preferences of Culex restuans and Culex pipiens by using ovitraps and gravid traps baited with selected infusions. For the ovitrap collections, 4 different infusions (manure, hay, grass, and rabbit chow) were used. Significant differences among infusions were detected on most sample dates for both species. For 3 of the first 4 wk of collections, the manure infusion collected significantly more Cx. restuans than all the other infusions. The hay and grass infusions collected the majority of the egg rafts during weeks 5-9. Cx. pipiens egg rafts were absent from the first 3 wk of collections. Of the remaining 6 wk, 4 showed significant differences in attractiveness of infusions, with the hay and grass infusions preferred by Cx. pipiens. Two infusions, manure and hay, were used for the gravid trap experiment and both Cx. restuans and Cx. pipiens data were combined for analysis. Only the first 2 wk showed significance, with manure being preferred over hay in both weeks. In later collections, the relative attractiveness of the hay infusion increased. A seasonal shift in infusion preference may be related to incubation temperature during preparation of the infusions. New infusions were prepared each week and incubation was done outside. Increased attractiveness of the hay infusion coincided with higher average temperatures in July and August. Hay infusion was very effective for trapping both Cx. pipiens and Cx. restuans in southwestern Virginia and is more convenient to use than manure. However, cool outside temperatures in the early season may interfere with the fermentation process and thus incubation should be done for a longer time or brought indoors.

  4. Molecular expression of aquaporin mRNAs in the northern house mosquito, Culex pipiens.

    PubMed

    Yang, Liu; Piermarini, Peter M

    2017-01-01

    Aquaporins (AQPs) are a family of membrane-bound proteins, originally described as water-channels, that broadly exist in prokaryotic and eukaryotic cells. However, some AQPs can transport small molecules (e.g. urea, glycerol) along with or preferentially to water. Previous work in the mosquitoes Aedes aegypti and Anopheles gambiae have characterized the molecular expression of one or more AQP genes and shown that they are involved in water homeostasis after blood feeding, tolerance to dehydration and heat stresses, and development of Plasmodium falciparum in the mosquito. However, the molecular expression of AQPs in the northern house mosquito Culex pipiens, the primary vector of West Nile virus, is presently unknown. Here we characterize the relative mRNA expression of six AQP genes in C. pipiens. We used quantitative RT-PCR (qPCR) to compare AQP mRNA levels 1) among different life stages and sexes, 2) between blood fed and non-blood fed adult females, 3) between adult females injected with phosphate-buffered saline (PBS) and non-injected females, and 4) between adult females exposed to a low vs. high relative humidity. We found that: 1) four AQP mRNAs (Drip, Bib, Eglp1, Aqp12L) were differentially expressed between larval and adult life stages, but not between sexes; 2) after a blood meal, the expression of two AQP mRNAs (Drip and Aqp12L) were down-regulated and one AQP mRNA (Eglp2) was up-regulated; 3) the expression of two AQP mRNAs (Drip and Prip) were down-regulated and one AQP mRNA (Eglp1) was up regulated after injecting the hemolymph of mosquitoes with PBS; and 4) the expression of Prip and Eglp2 mRNAs were both down-regulated in response to a low relative humidity. Our study is the first to characterize the molecular expression patterns of AQPs in C. pipiens, thus providing a foundation for future research elucidating the physiological and functional roles of AQPs in this important disease vector. Copyright © 2016 Elsevier Ltd. All rights reserved.

  5. 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

  6. High-density linkage maps fail to detect any genetic component to sex determination in a Rana temporaria family.

    PubMed

    Brelsford, A; Rodrigues, N; Perrin, N

    2016-01-01

    Sex chromosome differentiation in Rana temporaria varies strikingly among populations or families: whereas some males display well-differentiated Y haplotypes at microsatellite markers on linkage group 2 (LG2), others are genetically undistinguishable from females. We analysed with RADseq markers one family from a Swiss lowland population with no differentiated sex chromosomes, and where sibship analyses had failed to detect any association between the phenotypic sex of progeny and parental haplotypes. Offspring were reared in a common tank in outdoor conditions and sexed at the froglet stage. We could map a total of 2177 SNPs (1123 in the mother, 1054 in the father), recovering in both adults 13 linkage groups (= chromosome pairs) that were strongly syntenic to Xenopus tropicalis despite > 200 My divergence. Sexes differed strikingly in the localization of crossovers, which were uniformly distributed in the female but limited to chromosome ends in the male. None of the 2177 markers showed significant association with offspring sex. Considering the very high power of our analysis, we conclude that sex determination was not genetic in this family; which factors determined sex remain to be investigated.

  7. 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.

  8. Environmental and Biological Factors Influencing Culex pipiens quinquefasciatus (Diptera: Culicidae) Vector Competence for West Nile Virus

    PubMed Central

    Richards, Stephanie L.; Lord, Cynthia C.; Pesko, Kendra N.; Tabachnick, Walter J.

    2010-01-01

    Interactions between environmental and biological factors affect the vector competence of Culex pipiens quinquefasciatus for West Nile virus. Three age cohorts from two Cx. p. quinquefasciatus colonies were fed blood containing a low- or high-virus dose, and each group was held at two different extrinsic incubation temperatures (EIT) for 13 days. The colonies differed in the way that they responded to the effects of the environment on vector competence. The effects of mosquito age on aspects of vector competence were dependent on the EIT and dose, and they changed depending on the colony. Complex interactions must be considered in laboratory studies of vector competence, because the extent of the genetic and environmental variation controlling vector competence in nature is largely unknown. Differences in the environmental (EIT and dose) and biological (mosquito age and colony) effects from previous studies of Cx. p. quinquefasciatus vector competence for St. Louis encephalitis virus are discussed. PMID:20595491

  9. The susceptibility of Culex pipiens fatigans larvae to insecticides in Rangoon, Burma

    PubMed Central

    Rosen, Philip

    1967-01-01

    Since April 1963, the susceptibility to insecticides of larvae of Culex pipiens fatigans has been extensively studied by the WHO Filariasis Research Unit in Rangoon, Burma, as part of a programme of research aimed at establishing which compound or group of compounds could be used to control the vector of filariasis. Larvae from various localities in Rangoon showed wide variations in susceptibility to chlorinated hydrocarbons. There were also seasonal variations and it was difficult to standardize test conditions. Nevertheless, resistance to this group of compounds, especially DDT, sufficient to prevent control has been clearly demonstrated. As C. p. fatigans has never been subjected to intensive control by insecticides in Rangoon, the results confirm the findings made elsewhere that this mosquito is naturally resistant to chlorinated hydrocarbons. The larvae are, however, susceptible to organophosphorus compounds, especially fenthion and parathion. PMID:4230023

  10. 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.

  11. 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.

  12. 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.

  13. Population

    EPA Science Inventory

    Population growth influences many stressors on Narragansett Bay and its Watershed, including all landscape and chemical stressors discussed in other chapters of this report. In numerous ways, population growth affects the condition of the Bay ecosystem, Watershed ecosystem, and h...

  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. 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 ...

  16. [Population].

    PubMed

    1979-01-01

    Data on the population of Venezuela between 1975 and 1977 are presented in descriptive tables and graphs. Information is included on the employed population according to category, sex, and type of economic activity, and by sex, age, and area on the employment rate and the total, the economically active, and the unemployed population.

  17. 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…

  18. 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.

  19. Insecticidal activity of isobutylamides derived from Piper nigrum against adult of two mosquito species, Culex pipiens pallens and Aedes aegypti.

    PubMed

    Park, Il-Kwon

    2012-01-01

    The insecticidal activity of Piper nigrum fruit-derived piperidine alkaloid (piperine) and N-isobutylamide alkaloids (pellitorine, guineensine, pipercide and retrofractamide A) against female adults of Culex pipiens pallens and Aedes aegypti was examined. On the basis of 24-h LD(50) values, the compound most toxic to female C. pipiens pallens was pellitorine (0.4 µg/♀) followed by guineensine (1.9 µg/♀), retrofractamide A (2.4 µg/♀) and pipercide (3.2 µg/♀). LD(50) value of chlorpyrifos was 0.03 µg/♀. Against female A. aegypti, the insecticidal activity was more pronounced in pellitorine (0.17 µg/♀) than in retrofractamide A (1.5 µg/♀), guineensine (1.7 µg/♀), and pipercide (2.0 µg/♀). LD(50) value of chlorpyrifos was 0.0014 µg/♀.

  20. Surveys for presence of Oregon spotted frog (Rana pretiosa): background information and field methods

    USGS Publications Warehouse

    Pearl, Christopher A.; Clayton, David; Turner, Lauri

    2010-01-01

    The Oregon spotted frog (Rana pretiosa) is the most aquatic of the native frogs in the Pacific Northwest. The common name derives from the pattern of black, ragged-edged spots set against a brown or red ground color on the dorsum of adult frogs. Oregon spotted frogs are generally associated with wetland complexes that have several aquatic habitat types and sizeable coverage of emergent vegetation. Like other ranid frogs native to the Northwest, Oregon spotted frogs breed in spring, larvae transform in summer of their breeding year, and adults tend to be relatively short lived (3-5 yrs). Each life stage (egg, tadpole, juvenile and adult) has characteristics that present challenges for detection. Breeding can be explosive and completed within 1-2 weeks. Egg masses are laid in aggregations, often in a few locations in large areas of potential habitat. Egg masses can develop, hatch, and disintegrate in <2 weeks during warm weather. Tadpoles can be difficult to identify, have low survival, and spend most of their 3-4 months hidden in vegetation or flocculant substrates. Juveniles and adults are often difficult to capture and can spend summers away from breeding areas. Moreover, a substantial portion of extant populations are of limited size (<100 breeding adults), and field densities of all life stages are often low. An understanding of the biology of the species and use of multiple visits are thus important for assessing presence of Oregon spotted frogs. This report is meant to be a resource for USDA Region 6 Forest Service (FS) and OR/WA Bureau of Land Management (BLM) personnel tasked with surveying for the presence of Oregon spotted frogs. Our objective was to summarize information to improve the efficiency of field surveys and increase chances of detection if frogs are present. We include overviews of historical and extant ranges of Oregon spotted frog. We briefly summarize what is known of Oregon spotted frog habitat associations and review aspects of behavior and

  1. 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

  2. The effects of Nosema algerae Vavra and undeen (Microsporida: Nosematidae) pathogen of Culex pipiens L.(Diptera: Culicidae) from Egypt on fecundity and longevity of the host.

    PubMed

    Seif, A I

    2000-12-01

    A microsporidian, Nosema algerae Vavra and undeen, was found parasitizing larvae and adults of a laboratory colony of Culex pipiens L. originated from Gharbia Governorate. A detailed examination of the developmental stages of the pathogen under light and electron microscopes showed that they are typical of the original characteristics of N. algerae. Other observations on the infected individuals revealed that spores of the pathogen were found in all organs of the infected mosquitoes except the nervous system. The most heavily infected organ was the alimentary canal, particularly the mid gut. The susceptibility of the different larval instars of a laboratory maintained Cx. pipiens colony to infection by N. algerae was determined. Using 24 hours exposure to a range of doses between 40 and 5 x 10(5) spores per cm, larvae of the 1st and 2nd instars were more susceptible to infection than 3rd and 4th instar larvae as indicated by the differences in the estimated IC50 dosages. The dosages of N. algerae which produced 50% mortality (LC50) in each group of treated larvae was approximately 25-30 times higher than the doses that caused 50% infection. When a sublethal dose of 1.2 x 10(3) spores per cm2 of N. algerae was used to induce infection in each of the different larval instars infection rates of 100% were obtained in all exposed larval instars and in adults developed from them. Females from larvae infected with N. algerae had significantly reduced fecundity, fertility, and longevity regardless the larval instar in which infection was initiated. However, the reduction was obviously high when 1st and 2nd instars were exposed. The accumulative effects of reduced survival and fecundity on the reproductive potential of the infected females derived from larvae that were infected as 1st and 2nd instars probably serve to limit the natural increase of mosquito populations. Further studies are necessary, however, to determine the efficacy of this pathogen on target species in

  3. Subtle effects of environmental stress observed in the early life stages of the Common frog, Rana temporaria

    PubMed Central

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

    2017-01-01

    Worldwide amphibian populations are declining due to habitat loss, disease and pollution. Vulnerability to environmental contaminants such as pesticides will be dependent on the species, the sensitivity of the ontogenic life stage and hence the timing of exposure and the exposure pathway. Herein we investigated the biochemical tissue ‘fingerprint’ in spawn and early-stage tadpoles of the Common frog, Rana temporaria, using attenuated total reflection-Fourier-transform infrared (ATR-FTIR) spectroscopy with the objective of observing differences in the biochemical constituents of the respective amphibian tissues due to varying water quality in urban and agricultural ponds. Our results demonstrate that levels of stress (marked by biochemical constituents such as glycogen that are involved in compensatory metabolic mechanisms) can be observed in tadpoles present in the pond most impacted by pollution (nutrients and pesticides), but large annual variability masked any inter-site differences in the frog spawn. ATR-FTIR spectroscopy is capable of detecting differences in tadpoles that are present in selected ponds with different levels of environmental perturbation and thus serves as a rapid and cost effective tool in assessing stress-related effects of pollution in a vulnerable class of organism. PMID:28317844

  4. DDTs in rice frogs (Rana limnocharis) from an agricultural site, South China: tissue distribution, biomagnification, and potential toxic effects assessment.

    PubMed

    Wu, Jiang-Ping; Zhang, Ying; Luo, Xiao-Jun; Chen, She-Jun; Mai, Bi-Xian

    2012-04-01

    Contamination with agricultural pesticides such as dichlorodiphenyltrichloroethane (DDT) and its metabolites, dichlorodiphenyldichloroethylene (DDE) and dichlorodiphenyldichloroethane (DDD), is among several proposed stressors contributing to the global declines in amphibian populations and species biodiversity. These chemicals were examined in insects and in the muscle, liver, and eggs of rice frogs (Rana limnocharis) from the paddy fields of an agricultural site in South China. The ΣDDT (sum of DDT, DDE, and DDD) concentrations ranged from 154 to 915, 195 to 1,400, and 165 to 1,930 ng/g lipid weight in the muscle, liver, and eggs, respectively. All the DDTs (DDT, DDE, and DDD) showed higher affinity for the liver relative to muscle tissue and can be maternally transferred to eggs in female frogs. The average biomagnification factors for DDTs ranged from 1.6 to 1.9 and 1.5 to 2.9 in female and male frogs, respectively, providing clear evidence of their biomagnification from insects to frogs. Compared with the reported DDT levels demonstrated to have toxic effects on frogs, DDTs in the present frogs are unlikely to constitute an immediate health risk. However, the adverse impacts of high DDT residues in eggs on the hatching success and their potential toxicity to the newly metamorphosed larval frogs should be assessed further.

  5. Endocrine-disrupting effects and reproductive toxicity of low dose MCLR on male frogs (Rana nigromaculata) in vivo.

    PubMed

    Jia, Xiuying; Cai, Chenchen; Wang, Jia; Gao, Nana; Zhang, Hangjun

    2014-10-01

    Toxic cyanobacterial blooms are potential global threats to aquatic ecosystems and human health. The World Health Organization has set a provisional guideline limit of 1 μg/L microcystin-LR (MCLR) in freshwater. However, MCLR concentrations in several water bodies have exceeded this level. Despite this recommended human safety standard, MCLR-induced endocrine-disrupting effects and reproductive toxicity on male frog (Rana nigromaculata) were demonstrated in this study. Results showed that sperm motility and sperm count were significantly and negatively correlated with exposure time and concentration. By contrast, abnormal sperm rate was positively correlated with both parameters. Ultrastructural observation results revealed abnormal sperm morphologies, vacuoles in spermatogenic cells, cell dispersion, incomplete cell structures, and deformed nucleoli. These results indicated that MCLR could induce toxic effects on the reproductive system of frogs, significantly decrease testosterone content, and rapidly increase estradiol content. Prolonged exposure and increased concentration enhanced the relative expression levels of P450 aromatase and steroidogenic factor 1; thus, endocrine function in frogs was disrupted. This study is the first to demonstrate in vivo MCLR toxicity in the reproductive system of male R. nigromaculata. This study provided a scientific basis of the global decline in amphibian populations.

  6. 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.

  7. 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.

  8. Disease associated with integumentary and cloacal parasites in tadpoles of northern red-legged frog Rana aurora aurora.

    PubMed

    Nieto, Nathan C; Camann, Michael A; Foley, Janet E; Reiss, John O

    2007-10-31

    A total of 6830 northern red-legged frog Rana aurora aurora tadpoles were examined under a dissecting microscope for oral disc, integumentary, and cloacal abnormalities in 13 ponds in and near Redwood National Park in northern California. Of these, 163 tadpoles were collected for histopathological investigation, including 115 randomly collected individuals, 38 collected with oral disc abnormalities, and 10 collected due to severe morbidity of unknown etiology. The tadpoles were infected with 8 parasites, including Batrachochytrium dendrobatidis (the amphibian chytrid), trematodes, leeches, and protozoa. Chytridiomycosis was detected at an overall prevalence of 6.4%, but prevalence was higher in tadpoles with oral disc lesions than in those with normal oral discs (43.5% versus 6.1%). Interestingly, infection was associated with some environmental and co-infection risk factors. Individual tadpoles possessed 0 to 5 species of parasites in varying intensities. Apiosoma sp. was the most prevalent (66%) and widespread. Tadpoles infected with B. dendrobatidis had a lower diversity of oral parasites than those uninfected. During the field portion of the study, a large number (approximately 500) of moribund and dead tadpoles was seen occurring at multiple locations within and surrounding Redwood National Park. Ten animals were collected for histological examination and a diverse protozoal infection was discovered, including some known pathogens of fish. This study is the first reporting parasitism and disease in natural populations of northern red-legged frogs.

  9. 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

  10. Evaluation of Some Plant Fruit Extracts for the Control of West Nile Virus Vector Culex pipiens (Diptera: Culicidae).

    PubMed

    Koc, Samed; Evren, Ozay Hasan; Cetin, Huseyin

    2016-12-01

    The extracts of different parts of plants were found very effective against various pests. The aim of this research was to determine the insecticidal activity of fruit methanol extracts obtained from Melia azedarach (Meliaceae), Phoenix theophrasti (Arecaceae), Styphnolobium japonicum (Fabaceae) and Pyracantha coccinea (Rosaceae) against the larvae of Culex pipiens (Diptera: Culicidae). The fruits of test plants were collected from the Campus of Akdeniz University, Antalya, Turkey in 2013. A series of concentrations of the extracts ranging from 62.5-1000 ppm were tested against second instar larvae. Only the extracts of Me. azedarach and Ph. theoprasti showed significant larvicidal activity against Cx. pipiens and the LC50 values of these extracts were found to be 169.48 and 220.60 ppm, respectively. This is the first research investigating the insecticidal or larvicidal activity of Ph. theophrasti, St. japonicum and Py. coccinea extracts on mosquitoes. The methanol extract of fruits of Me. azedarach and Ph. theophrasti showed significantly higher larvicidal activity against Cx. pipiens.

  11. Rainfall Influences Survival of Culex pipiens (Diptera: Culicidae) in a Residential Neighborhood in the mid-Atlantic USA

    PubMed Central

    Jones, Christy E.; Lounibos, L. Philip; Marra, Peter P.; Kilpatrick, A. Marm

    2012-01-01

    Measurement of the survival and dispersal rates of mosquito vectors is an important step in designing and implementing control strategies. Vector survival plays a key role in determining the intensity of pathogen transmission, and vector movement determines the spatial scale on which control efforts must operate to be effective. We provide the first estimates of field survival and dispersal rates for Culex pipiens in North America, an important enzootic and bridge vector for West Nile virus (WNV). We conducted mark-release-recapture studies in a residential area near Washington DC in two consecutive years and fit nonlinear regression models to the recapture data that incorporate weather information into survival and recapture probabilities. We found that daily survival rates were not significantly different between the two years but were negatively affected by rainfall. The daily survival rate was 0.904 ± 0.037 (SE), which implies an average longevity of 10.4 days. As with other vector-borne pathogens, the measured survival rate suggests that at our site the majority of WNV-infected Cx. pipiens mosquitoes may perish before becoming infectious (being able to transmit WNV to hosts). We found relatively little evidence of dispersal following the initial night after release. Our results suggest that transmission of WNV and other pathogens transmitted by Cx. pipiens may be highly local and they highlight the importance of factors that influence survival of mosquito vectors. PMID:22679852

  12. 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

  13. 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.

  14. Evaluation of Some Plant Fruit Extracts for the Control of West Nile Virus Vector Culex pipiens (Diptera: Culicidae)

    PubMed Central

    Koc, Samed; Evren, Ozay Hasan; Cetin, Huseyin

    2016-01-01

    Background: The extracts of different parts of plants were found very effective against various pests. The aim of this research was to determine the insecticidal activity of fruit methanol extracts obtained from Melia azedarach (Meliaceae), Phoenix theophrasti (Arecaceae), Styphnolobium japonicum (Fabaceae) and Pyracantha coccinea (Rosaceae) against the larvae of Culex pipiens (Diptera: Culicidae). Methods: The fruits of test plants were collected from the Campus of Akdeniz University, Antalya, Turkey in 2013. A series of concentrations of the extracts ranging from 62.5–1000 ppm were tested against second instar larvae. Results: Only the extracts of Me. azedarach and Ph. theoprasti showed significant larvicidal activity against Cx. pipiens and the LC50 values of these extracts were found to be 169.48 and 220.60 ppm, respectively. This is the first research investigating the insecticidal or larvicidal activity of Ph. theophrasti, St. japonicum and Py. coccinea extracts on mosquitoes. Conclusion: The methanol extract of fruits of Me. azedarach and Ph. theophrasti showed significantly higher larvicidal activity against Cx. pipiens. PMID:28032112

  15. 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.

  16. Endohelminth fauna of the marsh frog Rana ridibunda from Lake Hazar, Turkey.

    PubMed

    Saglam, Naim; Arikan, Hatice

    2006-10-27

    In this study, 236 marsh frogs Rana ridibunda collected from Lake Hazar (Elazig, Turkey) at 15 d intervals between March 2001 and February 2002 were examined for endohelminths; of these, 148 (62.71%) frogs were found to be infected with helminths. In total, 9 helminth species (3 trematodes, 5 nematodes and 1 acanthocephalan) were identified. We observed Gorgoderina vitelliloba (prevalence 2.97%) in the urinary bladder, Haematoloechus variegatus (4.66%) and Rhabdias bufonis (8.90%) in the lung, Pleurogenoides medians (1.69%), Oswaldocruzia filiformis (3.81 %) and Acanthocephalus ranae (26.27 %) in the small intestine, Neoxysomatium brevicaudatum (16.95%) and Cosmocercoides sp. (3.39%) in the large intestine, and Eustrongylides excisus (14.41%) in the body cavity and on,the stomach. No helminth was found in the spleen, kidney, gall bladder, liver, heart or muscle. Of the 9 helminth species identified, Acanthocephalus ranae (26.27 %) had the highest prevalence and abundance and Oswaldocruzia filiformis (8.33+/-4.09) had the highest mean intensity.

  17. 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. Copyright © 2016 Elsevier Masson SAS. All rights reserved.

  18. 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.

  19. 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.

  20. Host-feeding patterns of Culex pipiens and other potential mosquito vectors (Diptera: Culicidae) of West Nile virus (Flaviviridae) collected in Portugal.

    PubMed

    Osório, Hugo Costa; Zé-Zé, Líbia; Alves, Maria João

    2012-05-01

    The host blood-feeding patterns of mosquito vectors affects the likelihood of human exposure to zoonotic pathogens, including West Nile Virus (family Flaviviridae, genus Flavivirus, WNV). In Portugal, data are unavailable regarding the blood-feeding habits of common mosquito species, including Culex pipiens L., considered the primary vector of WNV to humans. The sources of bloodmeals in 203 blood-fed mosquitoes of nine species collected from June 2007 to November 2010 in 34 Portuguese counties were analyzed by sequencing cytochrome-b partial fragments. Cx. pipiens was the most common species collected and successfully analyzed (n = 135/78). In addition, blood-fed females of the following species were analyzed: Ochlerotatus caspius Pallas (n = 20), Culex theileri Theobald (n = 16), Anopheles maculipennis s.l. Meigen (n = 10), Culiseta longiareolata Macquart (n = 7), Aedes aegypti L. (n = 6), Culex perexiguus Theobald (n = 3), Culiseta annulata Schrank (n = 3), and Ochlerotatus detritus Haliday (n = 3). The Cx. pipiens mosquitoes fed predominantly on birds (n = 55/78, 70.5%), with a high diversity of avian species used as hosts, although human blood was identified in 18 specimens (18/78, 23.1%). No significant differences were found between the host-feeding patterns of blood-fed Cx. pipiens collected in residential and nonresidential habitats. The occurrence of human derived blood meals and the presence of a mix avian-human bloodmeal accordingly suggest this species as a potential vector of WNV. Therefore, in Portugal, Cx. pipiens may play a role both in the avian-to-avian enzootic WNV cycle and in the avian-to-mammal transmission. In this context, the identity of Cx. pipiens (considering the forms molestus and pipiens) and the potential consequence on feeding behavior and WNV transmission are discussed.

  1. Population.

    ERIC Educational Resources Information Center

    International Planned Parenthood Federation, London (England).

    In an effort to help meet the growing interest and concern about the prob