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Sample records for rana pipiens populations

  1. 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.

  2. 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.

  3. 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.

  4. 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.

  5. 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...

  6. 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

  7. 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.

  8. 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.

  9. 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.

  10. 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

  11. 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.

  12. 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

  13. 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.

  14. 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.

  15. 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.

  16. 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.

  17. 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.

  18. 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

  19. 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.

  20. 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

  1. 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

  2. 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.

  3. 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

  4. 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

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

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

  6. 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...

  7. 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.

  8. 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.

  9. 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.

  10. 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.

  11. 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

  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. 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.

  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. 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.

  16. 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.

  17. 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

  18. 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.

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

    PubMed Central

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

    2016-01-01

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

  20. 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.

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

    PubMed Central

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

    2013-01-01

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

  2. 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.

  3. 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...

  4. 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.

  5. 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

  6. 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

  7. 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.

  8. 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.

  9. 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...

  10. 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...

  11. 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

  12. 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.

  13. 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

  14. 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.

  15. 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

  16. 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

  17. 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.

  18. 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

  19. 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.

  20. 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

  1. 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.

  2. 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.

  3. 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.

  4. 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.

  5. 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

  6. Multiple duplications of the rare ace-1 mutation F290V in Culex pipiens natural populations.

    PubMed

    Alout, Haoués; Labbé, Pierrick; Berthomieu, Arnaud; Pasteur, Nicole; Weill, Mylène

    2009-12-01

    Two amino acid substitutions in acetylcholinesterase 1 (AChE1), G119S and F290V, are responsible for resistance to organophosphate and carbamate insecticides in Culex pipiens mosquitoes. These mutations generate very different levels of insensitivity to insecticide inhibitors. We described here a biochemical method that rapidly identifies AChE1 variants (susceptible, G119S and F290V, named S, R and V, respectively) present in individual mosquitoes. We investigated the frequency of AChE1 phenotypes in 41 field samples collected around the Mediterranean Sea. F290V substitution was found only in 15 samples and at low frequency, whereas G119S was highly spread in all samples. However, seven V distinct alleles were identified whereas only one R allele was present. The [V] enzymatic phenotype was never observed alone, and the V allele was always found associated with the susceptible and/or G119S AChE1 ([VS], [VR] or [VRS] phenotypes). Furthermore, we showed the presence of duplicated alleles, associating a susceptible and a V copy of the ace-1 gene, in most individuals analyzed for its presence. Evolutionary forces driving the large number of F290V ace-1 alleles and their low frequency in Mediterranean countries are discussed.

  7. 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.

  8. 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...

  9. 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.

  10. 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.

  11. 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

  12. 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.

  13. 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

  14. 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.

  15. 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.

  16. 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.

  17. 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.

  18. 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...

  19. 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.

  20. 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.

  1. 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

  2. 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.

  3. 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

  4. 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

  5. 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

  6. 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.

  7. 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...

  8. 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

  9. 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

  10. 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.

  11. 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.

  12. 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.

  13. 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.

  14. 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

  15. 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

  16. 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

  17. 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

  18. 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...

  19. 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

  20. 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.

  1. 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.

  2. 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.

  3. 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

  4. Diversification of Wolbachia endosymbiont in the Culex pipiens mosquito.

    PubMed

    Atyame, Célestine M; Delsuc, Frédéric; Pasteur, Nicole; Weill, Mylène; Duron, Olivier

    2011-10-01

    The α-proteobacteria Wolbachia are among the most common intracellular bacteria and have recently emerged as important drivers of arthropod biology. Wolbachia commonly act as reproductive parasites in arthropods by inducing cytoplasmic incompatibility (CI), a type of conditional sterility between hosts harboring incompatible infections. In this study, we examined the evolutionary histories of Wolbachia infections, known as wPip, in the common house mosquito Culex pipiens, which exhibits the greatest variation in CI crossing patterns observed in any insect. We first investigated a panel of 20 wPip strains for their genetic diversity through a multilocus scheme combining 13 Wolbachia genes. Because Wolbachia depend primarily on maternal transmission for spreading within arthropod populations, we also studied the variability in the coinherited Cx. pipiens mitochondria. In total, we identified 14 wPip haplotypes, which all share a monophyletic origin and clearly cluster into five distinct wPip groups. The diversity of Cx. pipiens mitochondria was extremely reduced, which is likely a consequence of cytoplasmic hitchhiking driven by a unique and recent Wolbachia invasion. Phylogenetic evidence indicates that wPip infections and mitochondrial DNA have codiverged through stable cotransmission within the cytoplasm and shows that a rapid diversification of wPip has occurred. The observed pattern demonstrates that a considerable degree of Wolbachia diversity can evolve within a single host species over short evolutionary periods. In addition, multiple signatures of recombination were found in most wPip genomic regions, leading us to conclude that the mosaic nature of wPip genomes may play a key role in their evolution.

  5. 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...

  6. 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.

  7. 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.

  8. 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.

  9. 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...

  10. 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.

  11. 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.

  12. 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

  13. 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.

  14. 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.

  15. 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...

  16. 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.

  17. 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.

  18. 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

  19. 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.

  20. 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.

  1. 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

  2. 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.

  3. [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

  4. 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.

  5. 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

  6. 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.

  7. 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.

  8. 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.

  9. 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.

  10. 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 ...

  11. 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

  12. 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

  13. 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

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

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

  15. 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.

  16. 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.

  17. 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

  18. 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.

  19. 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

  20. 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

  1. Do Frogs Still Get Their Kicks On Route 66? A Transcontinental Transect For Amphibian Chytrid Fungus (Batrachochytrium Dendrobatidis) Infection On U.S. Department Of Defense Installations

    DTIC Science & Technology

    2011-01-04

    samples during the spring/early summer sampling period (due to cold and snow ), when the majority of positive samples at other bases were collected (see... Leopard Frog (Rana pipiens) populations suggest intraspecies differences in resistance to pathogens. Develop Comp Immunol33: 1247-1257. Vredenburg VT...Woodhams DC, Hyatt AD, Boyle DG, Rollins-Smith LA (2007a) The Northern Leopard Frog Rana pipiens is a widespread reservoir species harboring

  2. 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

  3. 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.

  4. 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

  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. 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.

  7. 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?|

  8. 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.

  9. 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.

  10. 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.

  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. 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.

  13. 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.

  14. 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.

  15. 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.

  16. 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

  17. 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.

  18. 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.

  19. 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

  20. 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.

  1. 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...

  2. 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

  3. 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.

  4. 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...

  5. 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

  6. 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.

  7. 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

  8. 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

  9. 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.

  10. 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.

  11. 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.

  12. 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

  13. 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.

  14. 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.

  15. 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.

  16. 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

  17. 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.

  18. 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.

  19. High incidence of ace-1 duplicated haplotypes in resistant Culex pipiens mosquitoes from Algeria.

    PubMed

    Alout, Haoues; Labbé, Pierrick; Pasteur, Nicole; Weill, Mylène

    2011-01-01

    The status of genes conferring resistance to organophosphate and carbamate insecticides has been examined in Culex pipiens pipiens mosquitoes sampled in Algeria. Presence of overproduced esterases was sporadic, but acetylcholinesterase-1 resistant alleles were observed in almost all samples. We focused our study on the AChE1 G119S substitution characterized in almost all samples, mostly at the heterozygous state. A genetic test revealed the presence of ace-1 duplication associating a susceptible and a resistant ace-1 copy. Molecular characterization showed a high occurrence of ace-1 duplication with six distinct duplicated alleles out of four samples. The inferred frequency of duplicated allele suggests that it is replacing the single resistant G119S allele. Finally, we discuss the mechanism at the origin of these duplicated haplotypes and their consequences on the management of insecticide resistance.

  20. 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

  1. 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.

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

    PubMed Central

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

    2016-01-01

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

  3. 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.

  4. 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.

  5. 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.

  6. 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.

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

    PubMed

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

    2011-12-01

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

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

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

  9. [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.

  10. [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.

  11. 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.

  12. 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.

  13. 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.

  14. 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.

  15. 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.

  16. 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.

  17. 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.

  18. 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.

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

    PubMed Central

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

    2014-01-01

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

  20. 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.

  1. 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.

  2. 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.

  3. 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.

  4. 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.

  5. 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.

  6. 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.

  7. 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.

  8. 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.

  9. 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.

  10. 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.

  11. 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.

  12. 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.

  13. 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.

  14. 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%.

  15. 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

  16. 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.

  17. 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.

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

    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...

  19. 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

  20. 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.

  1. 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.

  2. 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

  3. 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.

  4. 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...

  5. 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.

  6. 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.

  7. 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.

  8. 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.

  9. 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.

  10. 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

  11. [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.

  12. 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.

  13. 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.

  14. 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.

  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. 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.

  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. [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.

  19. 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

  20. 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

  1. 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.

  2. 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.

  3. 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 problems created by the rapid growth of population, The International Planned Parenthood Federation has prepared this booklet with the aim of assisting the study of the history and future trends of population growth and its impact on individual and family welfare, national,…

  4. 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.

  5. 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.

  6. 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/♀.

  7. 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 ...

  8. 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

  9. 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.

  10. 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.

  11. 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

  12. 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.

  13. 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

  14. 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.

  15. 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.

  16. 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

  17. 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

  18. 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

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

    PubMed

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

    2016-06-01

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

  20. Sodium arsenite induced changes in survival, growth, metamorphosis and genotoxicity in the Indian cricket frog (Rana limnocharis).

    PubMed

    Singha, Utsab; Pandey, Neelam; Boro, Freeman; Giri, Sarbani; Giri, Anirudha; Biswas, Somava

    2014-10-01

    Arsenic contamination of the environment is a matter of great concern. Understanding the effects of arsenic on aquatic life will act as biological early warning system to assess how arsenic could shape the biodiversity in the affected areas. Rapid decline in amphibian population in recent decades is a cause of major concern. Over the years, amphibians have been recognized as excellent bio-indicators of environmental related stress. In the present study, we examined the toxic and genotoxic effects of sodium arsenite in the tadpoles of the Indian cricket frog (Rana limnocharis). Sodium arsenite at different concentrations (0, 50, 100, 200 and 400 μg L(-1)) neither induced lethality nor significantly altered body weight at metamorphosis. However, it accelerated the rate of metamorphosis at higher concentrations, reduced body size (snout-vent length) and induced developmental deformities such as loss of limbs. Besides, at concentration ranges between 100 and 400 μg L(-1), sodium arsenite induced statistically significant genotoxicity at 24, 48, 72 and 96 h of the exposure in a concentration-dependent manner. However, it did not show time effects as the highest frequency was found between 48 and 72 h which remained steady subsequently. The genotoxicity was confirmed by comet assay in the whole blood cells. These findings suggest that arsenic at environmentally relevant concentrations has significant sub-lethal effects on R.limnocharis, which may have long-term fitness consequence to the species and may have similar implications in other aquatic life too.

  1. 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

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

    SciTech Connect

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

    2008-09-15

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

  3. 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.

  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. 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.

  7. 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.

  8. 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.

  9. Solid-state NMR reveals differential carbohydrate utilization in diapausing Culex pipiens

    NASA Astrophysics Data System (ADS)

    Chang, James; Singh, Jugeshwar; Kim, Sungshil; Hockaday, William C.; Sim, Cheolho; Kim, Sung Joon

    2016-11-01

    Culex pipiens is the mosquito that vectors West Nile Virus and other human-pathogenic flavivruses in North America. In response to shortened day length and lower temperatures, female Cx. pipiense prepares for the diapause by actively feeding on carbohydrates to increase the biosynthesis of glycogen and lipid to store energy for overwintering. The effect of feeding different carbohydrates on glycogen and lipid biosynthesis in diapausing mosquitoes was investigated in vivo using 13C solid-state NMR. Diapause-destined adult females and nondiapausing counterparts after adult eclosion were fed with three different carbohydrate sources for 7 days: 1) 10% sucrose, 2) 10% D-[13C6]glucose, and 3) 1% D-[13C6]glucose co-provisioned with 10% sucrose. NMR measurements show that sucrose and glucose are metabolized differently in diapausing mosquitoes. Mosquitoes fed on sucrose primarily accumulate glycogen with increased branching structures, but less of lipids. In contrast, mosquitoes fed exclusively on glucose show accumulation of both glycogen and lipid with increased aliphatic chain length. Glucose is exclusively metabolized for the biosynthesis of triacylglyceride when mosquitoes were co-fed with sucrose. Our findings provide novel insights into the insect carbohydrate metabolism that governs glycogen and lipid biosynthesis during diapause, which is fundamental for the insect survival during inimical environments.

  10. Attract-and-kill strategy. Laboratory studies on hatched larvae of Culex pipiens.

    PubMed

    Michaelakis, Antonios; Mihou, Anastasia P; Koliopoulos, George; Couladouros, Elias A

    2007-10-01

    The attract-and-kill strategy is a new pest management technique that presupposes the intelligent combination of an attracting agent (e.g. pheromone) and a killing agent (e.g. insecticide). In the present study, the potential combination of the microencapsulated synthetic oviposition pheromone 6-acetoxy-5-hexadecanolide with an insecticide has been tested. Initially, polyurea microcapsules containing 6-acetoxy-5-hexadecanolide, the synthetic mixture of diastereomers of the oviposition pheromone of the mosquito species Culex quinquefasciatus Say (Diptera: Culicidae), were studied. Laboratory bioassays were performed to confirm the bioactivity of the microencapsulated pheromone on the oviposition activity of Culex pipiens L. biotype molestus Førskal (Diptera: Culicidae) with the aim of determining the optimum dose for oviposition response. Its effect was dose dependent, revealing an optimum dose of 300 mg of dried microcapsules. Attractancy over time was also studied. The microencapsulated pheromone was found to be sufficiently attractive to gravid female mosquitoes for a period of 40 days. Finally, the combination of the synthetic pheromone with the control agent temephos showed both an acceptable oviposition activity and sufficient larvicidal effect.

  11. Effect of chlorfenapyr on cypermethrin-resistant Culex pipiens pallens Coq mosquitoes.

    PubMed

    Yuan, J Z; Li, Q F; Huang, J B; Gao, J F

    2015-03-01

    Chlorfenapyr is a promising pyrrole insecticide with a unique mechanism of action that does not confer cross-resistance to neurotoxic insecticides. The effect of chlorfenapyr on pyrethorid-resistant Culex pipiens pallens Coq (Diptera: Culicidae) has not been fully investigated under laboratory conditions. In this study, cypermethrin-resistant C. p. pallens exhibited 376.79-fold and 395.40-fold increase in resistance to cypermethrin compared with susceptible strains after exposure for 24 and 48h, respectively. Larvae and adults were tested for susceptibility using dipping, topical, and impregnated paper methods as recommended by the WHO. No cross-resistance to chlorfenapyr was found. Increased mortality was apparent between 48 and 72h, indicating a slow rate of toxic activity. Synergism experiments with piperonyl butoxide (PBO) showed an antagonistic effect on chlorfenapyr toxicity. Mixtures of chlorfenapyr and cypermethrin could therefore provide additional benefits over either insecticide used alone. Mixtures of 5ng/ml chlorfenapyr and 500ng/ml cypermethrin exhibited a slight synergistic effect on cypermethrin-resistant mosquitoes (3.33, 6.84 and 2.34% after 24, 48 and 72h exposure, respectively. This activity was lost when the chlorfenapyr concentration was increased to 10 or 20ng/ml. Chlorfenapyr showed quite good results for pyrethroid-resistant C. p. pallens, and could improve public health by reducing the occurrence of mosquito bites and subsequently protecting against transmission of lymphatic filariasis and Japanese encephalitis.

  12. Larvicidal activity of extracts of Ginkgo biloba exocarp for three different strains of Culex pipiens pallens.

    PubMed

    Sun, Lixin; Dong, Huiqin; Guo, Chongxia; Qian, Jin; Sun, Jing; Ma, Lei; Zhu, Changliang

    2006-03-01

    Ethanolic extracts from the Ginkgo biloba L. exocarp from the Chinese ginkgo were assayed against larvae of three strains of Culex pipiens pallens Coquillett. The chemical compositions were detected using a Hewlett-Packard 6890/5973 mass spectrometric detector. The larvicidal bioassay was carried out according to the recommendations of the World Health Organization. The analysis of the essential oil of ginkgo exocarp showed that its major components are ginkgo acid (85.3%) and ginkgo phenolic (5.69%). The larvicidal bioassay showed that extracts of ginkgo exocarp have LC50 of 18.6, 12.7, and 25.0 mg/liter for deltamethrin-susceptible, deltamethrin-resistant, and field strains, respectively. The acute toxicity concentrations of the ginkgo extracts that killed 50% (LD50) of Wistar rats within 2 wk and young carp within 96 h were 4947.2 mg/kg and 557.9 mg/liter, respectively. These results are promising in creating new, effective, and affordable approaches to mosquito control.

  13. Solid-state NMR reveals differential carbohydrate utilization in diapausing Culex pipiens

    PubMed Central

    Chang, James; Singh, Jugeshwar; Kim, Sungshil; Hockaday, William C.; Sim, Cheolho; Kim, Sung Joon

    2016-01-01

    Culex pipiens is the mosquito that vectors West Nile Virus and other human-pathogenic flavivruses in North America. In response to shortened day length and lower temperatures, female Cx. pipiense prepares for the diapause by actively feeding on carbohydrates to increase the biosynthesis of glycogen and lipid to store energy for overwintering. The effect of feeding different carbohydrates on glycogen and lipid biosynthesis in diapausing mosquitoes was investigated in vivo using 13C solid-state NMR. Diapause-destined adult females and nondiapausing counterparts after adult eclosion were fed with three different carbohydrate sources for 7 days: 1) 10% sucrose, 2) 10% D-[13C6]glucose, and 3) 1% D-[13C6]glucose co-provisioned with 10% sucrose. NMR measurements show that sucrose and glucose are metabolized differently in diapausing mosquitoes. Mosquitoes fed on sucrose primarily accumulate glycogen with increased branching structures, but less of lipids. In contrast, mosquitoes fed exclusively on glucose show accumulation of both glycogen and lipid with increased aliphatic chain length. Glucose is exclusively metabolized for the biosynthesis of triacylglyceride when mosquitoes were co-fed with sucrose. Our findings provide novel insights into the insect carbohydrate metabolism that governs glycogen and lipid biosynthesis during diapause, which is fundamental for the insect survival during inimical environments. PMID:27853281

  14. West Nile Virus Infection Alters Midgut Gene Expression in Culex pipiens quinquefasciatus Say (Diptera: Culicidae)

    PubMed Central

    Smartt, Chelsea T.; Richards, Stephanie L.; Anderson, Sheri L.; Erickson, Jennifer S.

    2009-01-01

    Alterations in gene expression in the midgut of female Culex pipiens quinquefasciatus exposed to blood meals containing 6.8 logs plaque-forming units/mL of West Nile virus (WNV) were studied by fluorescent differential display. Twenty-six different cDNAs exhibited reproducible differences after feeding on infected blood. Of these, 21 cDNAs showed an increase in expression, and 5 showed a decrease in expression as a result of WNV presence in the blood meal. GenBank database searches showed that one clone with increased expression, CQ G12A2, shares 94% identity with a leucine-rich repeat-containing protein from Cx. p. quinquefasciatus and 32% identity to Toll-like receptors from Aedes aegypti. We present the first cDNA clone isolated from female Cx. p. quinquefasciatus midgut tissue whose expression changes on exposure to WNV. This cDNA represents a mosquito gene that is an excellent candidate for interacting with WNV in Cx. p. quinquefasciatus and may play a role in disease transmission. PMID:19635880

  15. The duration of egg, larval and pupal stages of Culex pipiens fatigans in Rangoon, Burma*

    PubMed Central

    de Meillon, Botha; Sebastian, Anthony; Khan, Z. H.

    1967-01-01

    Laboratory experiments to determine the duration of the immature stages of Culex pipiens fatigans were carried out because such information is important from the point of view of control by larvicides. At a temperature of 25.1°C±0.7°C the mean incubation period is 27.11±0.57 hours. Females spend a longer time in the pupal stage than males (34.16±0.74 hours and 32.95±0.75 hours, respectively, at 28.6°C±0.8°C; there is no 24-hour pupating or emerging rhythm. The duration of larval life is longer for the female (135.3±4.4 hours) than for the male (118.4±2.4 hours). Larvae that take a long time to pupate also take a long time to emerge. Withholding of food for a few hours from first-stage larvae increases the duration of larval life but does not affect that of pupal life. These observations on the differences between the sexes in the duration of larval and pupal life are in agreement with observations made on Aedes aegypti in Uganda. PMID:4227199

  16. Insecticide resistance in Culex pipiens quinquefasciatus and Aedes albopictus mosquitoes from La Réunion Island.

    PubMed

    Tantely, Michaël Luciano; Tortosa, Pablo; Alout, Haoues; Berticat, Claire; Berthomieu, Arnaud; Rutee, Abdoul; Dehecq, Jean-Sébastien; Makoundou, Patrick; Labbé, Pierrick; Pasteur, Nicole; Weill, Mylène

    2010-04-01

    Resistance to insecticides was monitored on Culex pipiens quinquefasciatus mosquitoes collected in twelve localities of La Réunion, a geographically isolated island of the Indian Ocean. This mosquito is of medical concern in the region as a known vector for filariasis and a potential vector for West Nile and Rift Valley Fever viruses. Our bioassays indicated the presence of resistance to all tested insecticides, i.e. organochlorides, organophosphates and pyrethroids. A molecular investigation revealed a higher frequency of resistance genes in the coastal areas compared to elevated rural sites, probably reflecting the different nature of insecticide pressures together with the genetic cost of resistance alleles. A simple molecular test was developed to detect Rdl allele, encoding a gamma-aminobutyric acid (GABA) receptor resistant to dieldrin. Unexpectedly high Rdl frequencies were recorded over the whole island, despite this insecticide having been banned for over 15 years. This resistant allele was also detected for the first time in two samples of Aedes albopictus, a species recently involved in severe Chikungunya epidemics on the island. Rdl selection in these two mosquito species discloses current insecticide pressures in urban areas, from unknown origins, that should be taken into account to develop vector control strategies.

  17. Syritta pipiens (Diptera: Syrphidae), a new species associated with human cadavers.

    PubMed

    Magni, Paola A; Pérez-Bañón, Celeste; Borrini, Matteo; Dadour, Ian R

    2013-09-10

    The analyses of necrophagous insects feeding on a corpse can be successfully used to estimate the minimum time since death. A minimum time frame is sometimes an underestimate, but it is actually the only method that can provide such information when decomposed remains are found at a crime scene. Many insects are known to be colonisers of a corpse, but because there is an endless spectrum of crime scene environments, the development data bases for necrophagous insects is incomplete. The two cases detailed in this paper show different entomological patterns due to the different environments (well and burial) and locations (south and central Italy) where the two cadavers were found. Common to both of these cases' was the discovery of the corpse in the same period of the year (January) and the presence of Syritta pipiens (Diptera: Syrphidae), a species that has never been associated with deceased humans. The ecological information concerning this insect was used in combination with the more typical entomofauna found on the corpse to provide a minimum post mortem interval.

  18. Studies on the Mechanism of DDT Resistance in Culex pipiens fatigans

    PubMed Central

    Kalra, R. L.

    1970-01-01

    The mechanism of DDT resistance in Culex pipiens fatigans is poorly understood. Earlier studies indicated that the dehydrochlorination of DDT does not explain resistance in this species. Studies on the role of lipids as a mechanism of resistance included the estimation of lipid content and the determination of the proportions of different classes of lipids in the larvae of susceptible and resistant strains. There was no evidence of any correlation between the lipid content and DDT resistance in this species and the proportions of neutral lipids, phospholipids and fatty acids of different strains did not indicate any consistent correlation with DDT resistance. Within one strain, the larvae containing the higher amount of lipids were able to resist better the toxic effect of DDT. Analysis of fatty acids of the larvae that survived and died as a result of treatment with DDT did not reveal any difference. Neither p,p′-DDT nor o,p′-DDT at sublethal concentration affected the lipids of the larvae of susceptible and resistant strains. PMID:5310957

  19. Effects of sublethal doses of DDT on the reproduction and susceptibility of Culex pipiens L.

    PubMed

    Zaghloul, T M; Brown, A W

    1968-01-01

    The effect of non-selecting doses of DDT on the induction of resistance was investigated in the house mosquito, Culex pipiens. Sublethal doses of DDT were applied to the adults in each generation, at levels which just fell short of causing mortality in these adults. A susceptible strain, a DDT-resistant strain, and a slightly DDT-tolerant strain were so treated for 6-7 generations.It was found that the treatment initially caused approximately 25% of the ovaries to degenerate, and reduced the proportion of females that fed and oviposited. This reduction in biotic potential became aggravated in successive generations of the DDT-resistant and DDT-tolerant strains, which failed to show any material increase in resistance level. In the susceptible strain, however, the biotic potential became enhanced, and considerable resistance was developed. It was concluded that the increase of resistance in this strain was due to hidden selection, of eggs in the ovary, and of females which failed to oviposit.

  20. Diurnal resting behavior of adult Culex pipiens in an arid habitat in Israel and possible control measurements with toxic sugar baits.

    PubMed

    Schlein, Yosef; Müller, Günter C

    2012-10-01

    The distribution of resting Culex pipiens s.l., L. in vegetation at the margins of breeding sites and the effects of a narrow, surrounding, sprayed belt of sugar, food dye and toxin on adult mosquitoes were studied near two pairs of control and experimental sewage ponds close to human habitation in the Judean hills. Control belts were without toxin. A sprayed belt of sugar and toxin 0.5 m from the water gradually reduced the population to an average of 38.3 mosquitoes per trap, 7.6% of the highest catch, which was 504.6 mosquitoes per trap in the control site. In the second experiment, in which bait belts were 5 m from the water, the toxic bait spraying was followed by a rise in catches from 207.9 to 274.9 mosquitoes. This was 41% of the 670.2 mosquitoes per trap in the parallel control site. In areas without toxin treatment, diurnal catches by net amounted to 20,705 mosquitoes. Of these, 86.1% (17,825) were caught within 1m of the water while only 8.2% (1701) were caught at a distance of 3 m. The remainders were caught up to 20 m away. Parity status was determined for female samples caught by net. In areas without toxin, parous females accounted for 37% of the catch and 13.2% were young, meconium containing specimens. The population diminished following spraying of toxic bait 1m from the water and included 13% parous females and 17.6% had meconium in the gut.

  1. Effect of mercuric chloride on fertilization and larval development in the River Frog, Rana heckscheri (Wright) (Anura: Ranidae)

    SciTech Connect

    Punzo, F. )

    1993-10-01

    Previous investigations have indicated that heavy metals such as copper, cadmium, lead and mercury can act as systemic toxicants in many species of wildlife. Although numerous studies have emphasized the effects of metals and pesticides on metabolism, growth, survivorship, neural processes and reproduction in a number of taxa, little information is available on the effects of sublethal concentrations of metals on the reproductive physiology of amphibians. Industrial processes and mining activities can release substantial concentrations of heavy metals such as mercury into aquatic habitats. Since most amphibians have obligate aquatic larval stages, they are exposed to pollutants discharged into the aquatic environment. Amphibians can act as accumulators of heavy metals and their larval stages are useful indicators of pollution levels in the field. What little data are available, indicate that metals can significantly reduce viability in amphibians through their actions on metabolism, development and gametogenesis. The recent concerns over worldwide declines in amphibian populations and the susceptibility of amphibian populations to environmental toxicants, led me to assess the effect of mercuric chloride, one of the most common and persistent toxicants in aquatic environments, on fertilization and larval development in the river frog, Rana heckscheri (Wright). Although there is some information on fish, very little data are available on the effects of mercury on fertilization in amphibians generally, and no published data exist for R. heckscheri. This species is a conspicuous component of the aquatic fauna of parts of the southeastern United States where mercury levels have increased significantly over the last two decades. 22 refs., 2 tabs.

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

    USGS Publications Warehouse

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

    2004-01-01

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

  3. Influence of Nitrate and Nitrite on Thyroid Hormone Responsive and Stress-Associated Gene Expression in Cultured Rana catesbeiana Tadpole Tail Fin Tissue

    PubMed Central

    Hinther, Ashley; Edwards, Thea M.; Guillette, Louis J.; Helbing, Caren C.

    2012-01-01

    Nitrate and nitrite are common aqueous pollutants that are known to disrupt the thyroid axis. In amphibians, thyroid hormone (TH)-dependent metamorphosis is affected, although whether the effect is acceleration or deceleration of this developmental process varies from study to study. One mechanism of action of these nitrogenous compounds is through alteration of TH synthesis. However, direct target tissue effects on TH signaling are hypothesized. The present study uses the recently developed cultured tail fin biopsy (C-fin) assay to study possible direct tissue effects of nitrate and nitrite. Tail biopsies obtained from premetamorphic Rana catesbeiana tadpoles were exposed to 5 and 50 mg/L nitrate (NO3–N) and 0.5 and 5 mg/L nitrite (NO2–N) in the absence and presence of 10 nM T3. Thyroid hormone receptor β (TRβ) and Rana larval keratin type I (RLKI), both of which are TH-responsive gene transcripts, were measured using quantitative real time polymerase chain reaction. To assess cellular stress which could affect TH signaling and metamorphosis, heat shock protein 30, and catalase (CAT) transcript levels were also measured. We found that nitrate and nitrite did not significantly change the level of any of the four transcripts tested. However, nitrate exposure significantly increased the heteroscedasticity in response of TRβ and RLKI transcripts to T3. Alteration in population variation in such a way could contribute to the previously observed alterations of metamorphosis in frog tadpoles, but may not represent a major mechanism of action. PMID:22493607

  4. Comparative Transcriptomics Reveals Key Gene Expression Differences between Diapausing and Non-Diapausing Adults of Culex pipiens

    PubMed Central

    Kang, David S.; Denlinger, David L.; Sim, Cheolho

    2016-01-01

    Diapause is a critical eco-physiological adaptation for winter survival in the West Nile Virus vector, Culex pipiens, but little is known about the molecular mechanisms that distinguish diapause from non-diapause in this important mosquito species. We used Illumina RNA-seq to simultaneously identify and quantify relative transcript levels in diapausing and non-diapausing adult females. Among 65,623,095 read pairs, we identified 41 genes with significantly different transcript abundances between these two groups. Transcriptome divergences between these two phenotypes include genes related to juvenile hormone synthesis, anaerobic metabolism, innate immunity and cold tolerance. PMID:27128578

  5. Complete mitochondrial genome of the Seoul frog Rana chosenica (Amphibia, Ranidae): comparison of R. chosenica and R. plancyi.

    PubMed

    Ryu, Shi Hyun; Hwang, Ui Wook

    2011-06-01

    Here, we have sequenced the complete mitochondrial genome of the Seoul frog Rana chosenica (Amphibia, Ranidae), which is known as a Korean endemic species. It is listed as a vulnerable species by IUCN Red List and also an endangered species in South Korea. The complete mitochondrial genome of R. chosenica consists of 18,357 bp. Its gene arrangement pattern was identical with those of other Rana frogs. We compared the mitochondrial genome of R. chosenica with that of the Peking frog Rana plancyi that has been known closely related to R. chosenica. Nucleotide sequence similarity between the two whole mitochondrial genomes was 95.7%, and the relatively low similarity seems to indicate that the two species are distinctly separated on the species level. The information of mitochondrial genome comparison of the two species was discussed in detail.

  6. 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. (R825867)

    EPA Science Inventory

    The perspectives, information and conclusions conveyed in research project abstracts, progress reports, final reports, journal abstracts and journal publications convey the viewpoints of the principal investigator and may not represent the views and policies of ORD and EPA. Concl...

  7. MiR-278-3p regulates pyrethroid resistance in Culex pipiens pallens

    PubMed Central

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

    2014-01-01

    MicroRNAs (miRNAs) regulate gene expression and biological processes including embryonic development, innate immunity and infection in many species. Emerging evidence indicates that miRNAs are involved in drug resistance. However, little is known about the relationship between the miRNAs and insecticide resistance in mosquitos. Here, we reported that conserved miR-278-3p and its target gene are critical for pyrethroid resistance in Culex pipiens pallens. We found that CYP6AG11 is the target of miR-278-3p, through bioinformatic analysis and experimental verification. The expression level of miR-278-3p was lower, whereas the level of CYP6AG11 was higher in deltamethrin-resistant strain, which were detected using qRT-PCR. We also found that CYP6AG11 was regulated by miR-278-3p via a specific target site with the 3′UTR by luciferase reporter assay. In addition, overexpression of CYP6AG11 in the mosquito C6/36 cells showed better prolification than the cells with empty vector when treated by deltamethrin at different concentrations. Moreover, the overexpression of miR-278-3p through microinjection led to a significant reduction in the survival rate, and the level of CYP6AG11 was simultaneously reduced. These results indicated that miR-278-3p could regulate the pyrethroid resistance through CYP6AG11. PMID:25420996

  8. High chlorpyrifos resistance in Culex pipiens mosquitoes: strong synergy between resistance genes

    PubMed Central

    Alout, H; Labbé, P; Berthomieu, A; Makoundou, P; Fort, P; Pasteur, N; Weill, M

    2016-01-01

    We investigated the genetic determinism of high chlorpyrifos resistance (HCR), a phenotype first described in 1999 in Culex pipiens mosquitoes surviving chlorpyrifos doses ⩾1 mg l−1 and more recently found in field samples from Tunisia, Israel or Indian Ocean islands. Through chlorpyrifos selection, we selected several HCR strains that displayed over 10 000-fold resistance. All strains were homozygous for resistant alleles at two main loci: the ace-1 gene, with the resistant ace-1R allele expressing the insensitive G119S acetylcholinesterase, and a resistant allele of an unknown gene (named T) linked to the sex and ace-2 genes. We constructed a strain carrying only the T-resistant allele and studied its resistance characteristics. By crossing this strain with strains harboring different alleles at the ace-1 locus, we showed that the resistant ace-1R and the T alleles act in strong synergy, as they elicited a resistance 100 times higher than expected from a simple multiplicative effect. This effect was specific to chlorpyrifos and parathion and was not affected by synergists. We also examined how HCR was expressed in strains carrying other ace-1-resistant alleles, such as ace-1V or the duplicated ace-1D allele, currently spreading worldwide. We identified two major parameters that influenced the level of resistance: the number and the nature of the ace-1-resistant alleles and the number of T alleles. Our data fit a model that predicts that the T allele acts by decreasing chlorpyrifos concentration in the compartment targeted in insects. PMID:26463842

  9. "Singing in the Tube"--audiovisual assay of plant oil repellent activity against mosquitoes (Culex pipiens).

    PubMed

    Adams, Temitope F; Wongchai, Chatchawal; Chaidee, Anchalee; Pfeiffer, Wolfgang

    2016-01-01

    Plant essential oils have been suggested as a promising alternative to the established mosquito repellent DEET (N,N-diethyl-meta-toluamide). Searching for an assay with generally available equipment, we designed a new audiovisual assay of repellent activity against mosquitoes "Singing in the Tube," testing single mosquitoes in Drosophila cultivation tubes. Statistics with regression analysis should compensate for limitations of simple hardware. The assay was established with female Culex pipiens mosquitoes in 60 experiments, 120-h audio recording, and 2580 estimations of the distance between mosquito sitting position and the chemical. Correlations between parameters of sitting position, flight activity pattern, and flight tone spectrum were analyzed. Regression analysis of psycho-acoustic data of audio files (dB[A]) used a squared and modified sinus function determining wing beat frequency WBF ± SD (357 ± 47 Hz). Application of logistic regression defined the repelling velocity constant. The repelling velocity constant showed a decreasing order of efficiency of plant essential oils: rosemary (Rosmarinus officinalis), eucalyptus (Eucalyptus globulus), lavender (Lavandula angustifolia), citronella (Cymbopogon nardus), tea tree (Melaleuca alternifolia), clove (Syzygium aromaticum), lemon (Citrus limon), patchouli (Pogostemon cablin), DEET, cedar wood (Cedrus atlantica). In conclusion, we suggest (1) disease vector control (e.g., impregnation of bed nets) by eight plant essential oils with repelling velocity superior to DEET, (2) simple mosquito repellency testing in Drosophila cultivation tubes, (3) automated approaches and room surveillance by generally available audio equipment (dB[A]: ISO standard 226), and (4) quantification of repellent activity by parameters of the audiovisual assay defined by correlation and regression analyses.

  10. High chlorpyrifos resistance in Culex pipiens mosquitoes: strong synergy between resistance genes.

    PubMed

    Alout, H; Labbé, P; Berthomieu, A; Makoundou, P; Fort, P; Pasteur, N; Weill, M

    2016-02-01

    We investigated the genetic determinism of high chlorpyrifos resistance (HCR), a phenotype first described in 1999 in Culex pipiens mosquitoes surviving chlorpyrifos doses ⩾1 mg l(-1) and more recently found in field samples from Tunisia, Israel or Indian Ocean islands. Through chlorpyrifos selection, we selected several HCR strains that displayed over 10 000-fold resistance. All strains were homozygous for resistant alleles at two main loci: the ace-1 gene, with the resistant ace-1(R) allele expressing the insensitive G119S acetylcholinesterase, and a resistant allele of an unknown gene (named T) linked to the sex and ace-2 genes. We constructed a strain carrying only the T-resistant allele and studied its resistance characteristics. By crossing this strain with strains harboring different alleles at the ace-1 locus, we showed that the resistant ace-1(R) and the T alleles act in strong synergy, as they elicited a resistance 100 times higher than expected from a simple multiplicative effect. This effect was specific to chlorpyrifos and parathion and was not affected by synergists. We also examined how HCR was expressed in strains carrying other ace-1-resistant alleles, such as ace-1(V) or the duplicated ace-1(D) allele, currently spreading worldwide. We identified two major parameters that influenced the level of resistance: the number and the nature of the ace-1-resistant alleles and the number of T alleles. Our data fit a model that predicts that the T allele acts by decreasing chlorpyrifos concentration in the compartment targeted in insects.

  11. [Role of the hypothalamus in the regulation of primary sleep in the frog Rana temporaria].

    PubMed

    Shilling, N V

    1980-01-01

    It has been demonstrated that in the frog Rana temporaria the anterior hypothalamus is involved into regulation of the depth of two forms of rest--one with plastic, the other with decreased muscle tone. Resting state with catatonic muscle activity is associated with activation of the posterior hypothalamus. Participation of the anterior hypothalamus in regulation of the resting state with the decreased tone of skeletal muscles may be taken as one of the indications that this form of rest plays the role of sleep in amphibians, being transformed during evolution of vertebrates into the sleep of poikilotherms.

  12. Effects of nitrate and ammonium on larvae of Rana temporaria from the Pyrenees.

    PubMed

    Oromí, Neus; Sanuy, Delfí; Vilches, Marcel

    2009-05-01

    In order to investigate the effects of nitrate and ammonium on the amphibians in a pasture zone of the Catalonian Pyrenees, larvae of Rana temporaria from several ponds were exposed to different concentrations of nitrate (0-500 mg/L) and ammonium (0-1.2 mg/L). High concentrations of nitrate in the water caused mortality and reduced larval size of R. temporaria, whereas no effects on larvae were observed in ammonium conditions. The results suggest that, if the levels of nitrate reach about 100 mg/L, the possibility of survival of R. temporaria larvae may be reduced.

  13. Effects of adenosine perfusion on the metabolism and contractile activity of Rana ridibunda heart.

    PubMed

    Lazou, A; Beis, I

    1987-01-01

    The effects of adenosine were examined on the isolated perfused heart of the frog Rana ridibunda. Adenosine produced negative chronotropic and inotropic effects on frog ventricle in a concentration-dependent manner. The effects of adenosine on cardiac metabolism were also investigated by measuring the tissue content of adenine nucleotides, lactate, pyruvate, adenosine and inorganic phosphate, during adenosine perfusion. Adenosine had no effect on the tissue content of metabolites. No net synthesis of adenine nucleotides was observed during perfusion with increasing concentrations of adenosine. Lactate output from the heart decreased significantly with adenosine perfusion. Correlation of adenosine effects on cardiac muscle with the effects of hypoxia are discussed.

  14. Preparation of Ecofriendly Formulations Containing Biologically Active Monoterpenes with Their Fumigant and Residual Toxicities against Adults of Culex pipiens

    PubMed Central

    Taktak, Nehad E. M.; Awad, Osama M.; Elfiki, Souraya A.; Abou El-Ela, Nadia E.

    2016-01-01

    Different mixtures of monoterpenes (ketone, alcohol, and alkene) were loaded on paper discs and wax and their knockdown activities were evaluated against Culex pipiens adults. Some individual monoterpenes were also evaluated by residual toxicity technique. Citronella oil as a reference was also loaded separately or in combination with monoterpenes on paper discs and wax. The ketone monoterpenes mixture (camphor, menthone, carvone, and fenchone) on paper discs was the most active (KT50 = 17.20 min) followed by ketone monoterpenes with citronella oil (KT50 = 20.79 min) and citronella oil alone (KT50 = 28.72 min). Wax formulations proved that the ketone and alcohol (geraniol, thymol, and menthol) monoterpenes gave the most activity as knockdown (KT50 = 31.79 and 43.39 min, resp.). Alcohol monoterpenes formulation recorded KT50 = 43.39 min. Residual activity of tested individual monoterpenes reported that the menthol was more toxic than camphor and camphene. Generally, this study suggests that the monoterpenes have the properties, which make them used as eco-friendly compounds in the control programs of Cx. pipiens adult. The use of paper discs is more applicable than wax in the adulticidal formulations. PMID:27891154

  15. Functional circadian clock genes are essential for the overwintering diapause of the Northern house mosquito, Culex pipiens

    PubMed Central

    Meuti, Megan E.; Stone, Mary; Ikeno, Tomoko; Denlinger, David L.

    2015-01-01

    The short day lengths of late summer are used to program the overwintering adult diapause (dormancy) of the Northern house mosquito, Culex pipiens. Here, we investigated the role of clock genes in initiating this diapause and asked whether the circadian cycling of clock gene expression persists during diapause. We provide evidence that the major circadian clock genes continue to cycle throughout diapause and after diapause has been terminated. RNA interference (RNAi) was used to knock down the core circadian clock genes and to then assess the impact of the various clock genes on the ability of females to enter diapause. RNAi directed against negative circadian regulators (period, timeless and cryptochrome2) caused females that were reared under diapause-inducing, short day conditions to avert diapause. In contrast, knocking down the circadian-associated gene pigment dispersing factor caused females that were reared under diapause-averting, long day conditions to enter a diapause-like state. Our results implicate the circadian clock in the initiation of diapause in C. pipiens. PMID:25653422

  16. Temporal occurrence and community structure of helminth parasites in southern leopard frogs, Rana sphenocephala, from north central Oklahoma.

    PubMed

    Vhora, M Suhail; Bolek, Matthew G

    2015-03-01

    Currently, little information is available about the temporal recruitment of helminth communities in amphibian hosts. We examined the helminth community structure and temporal recruitment of helminth parasites in southern leopard frogs, Rana sphenocephala. Specifically, we were interested in how host life history such as habitat, age and/or size, diet, sex, and temporal variation in abiotic factors (precipitation and temperature) were important in determining monthly infection patterns of helminth populations and communities in southern leopard frogs. From May to September 2011, 74 southern leopard frogs were collected from Teal Ridge in Stillwater Payne County, OK, USA. Sixty-nine (93 %) of 74 frogs were infected with 1 or more helminth species. During our collecting period, the average monthly temperature was lowest in May and highest in July, and monthly precipitation was highest in May and lowest during the first week of September. The component community consisted of 11 species of helminth, including 1 larval and 1 adult cestode, 2 larval and 3 adult trematodes, and 1 juvenile and 3 adult nematodes. Of the 1790 helminths recovered, 51 % (911) were nematodes, 47 % (842) were cestodes, and 2 % (37) were trematodes. There were significant differences in the total abundance and mean species richness of helminths acquired by skin contact or through frog diet in monthly component communities of southern leopard frogs. A positive correlation existed for percentage of all helminths acquired by skin contact and monthly precipitation (r = 0.94, P < 0.01). Conversely, a negative correlation existed for monthly precipitation and percentage of helminths acquired by diet (r = -0.94, P < 0.01). Our results indicate that abiotic conditions such as precipitation have a major influence on the avenues for and constraints on the transmission of helminths with life cycles associated with water/moisture or terrestrial intermediate/paratenic hosts and are important in structuring

  17. Upregulation of two actin genes and redistribution of actin during diapause and cold stress in the northern house mosquito, Culex pipiens

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Two actin genes cloned from Culex pipiens L. are upregulated during adult diapause. Though actins 1 and 2 were expressed throughout diapause, both genes were most highly expressed early in diapause. These changes in gene expression were accompanied by a conspicuous redistribution of polymerized acti...

  18. Host-feeding habits of Culex pipiens and Aedes albopictus (Diptera: Culicidae) collected at the urban and suburban residential areas of Japan.

    PubMed

    Sawabe, Kyoko; Isawa, Haruhiko; Hoshino, Keita; Sasaki, Toshinori; Roychoudhury, Sudipta; Higa, Yukiko; Kasai, Shinji; Tsuda, Yoshio; Nishiumi, Isao; Hisai, Nobuo; Hamao, Shoji; Kobayashi, Mutsuo

    2010-05-01

    To evaluate the vectorial capacity of mosquitoes for viruses in Japan, the host-feeding habits of the mosquitoes were analyzed by sequencing polymerase chain reaction-amplified fragments of the cytochrome b and 16S ribosomal RNA regions of the mitochondrial DNA of 516 mosquitoes of 15 species from seven genera that were collected from residential areas during 2003-2006. Culex pipiens L. and Aedes albopictus Skuse were the most commonly collected species in urban and suburban residential areas. Anautogenous Culex pipiens pallens Coquillett was distinguished from the autogenous Cx. pipiens form molestus Forskal using a polymerase chain reaction-based identification method. Both Cx. p. pallens and Cx. p. form molestus exhibited similar host-feeding habits, broadly preferring avian (50.0 and 42.5% of avian, respectively) and mammalian (38.6 and 45.0% of avian, respectively) hosts, such as tree sparrows, ducks, and humans. Conversely, Ae. albopictus exhibited a highly mammalophilic and anthropophilic feeding pattern, with 84.2% feeding on mammalian hosts and 68.5% of these on humans. We concluded that in Japan, Cx. pipiens might play a significant role in the avian-to-mammal transmission of viruses, such as West Nile virus, whereas Ae. albopictus might play a role in the human-human transmission of dengue and Chikungunya viruses.

  19. Rangewide phylogeography and landscape genetics of the Western U.S. endemic frog Rana boylii (Ranidae): Implications for the conservation of frogs and rivers

    USGS Publications Warehouse

    Lind, A.J.; Spinks, P.Q.; Fellers, G.M.; Shaffer, H.B.

    2011-01-01

    Genetic data are increasingly being used in conservation planning for declining species. We sampled both the ecological and distributional limits of the foothill yellow-legged frog, Rana boylii to characterize mitochondrial DNA (mtDNA) variation in this declining, riverine amphibian. We evaluated 1525 base pairs (bp) of cytochrome b and ND2 fragments for 77 individuals from 34 localities using phylogenetic and population genetic analyses. We constructed gene trees using maximum likelihood and Bayesian inference, and quantified genetic variance (using AMOVA and partial Mantel tests) within and among hydrologic regions and river basins. Several moderately supported, geographically-cohesive mtDNA clades were recovered for R. boylii. While genetic variation was low among populations in the largest, most inclusive clade, samples from localities at the edges of the geographic range demonstrated substantial genetic divergence from each other and from more central populations. Hydrologic regions and river basins, which represent likely dispersal corridors for R. boylii, accounted for significant levels of genetic variation. These results suggest that both rivers and larger hydrologic and geographic regions should be used in conservation planning for R. boylii. ?? 2010 US Government.

  20. Comparison of diet, reproductive biology, and growth of the pig frog (Rana grylio) from harvested and protected areas of the Florida Everglades

    USGS Publications Warehouse

    Ugarte, C.A.; Rice, K.G.; Donnelly, M.A.

    2007-01-01

    Distinct differences in body size exist among three Rana grylio populations in areas of the Florida Everglades that differ in frog harvest pressure and hydroperiod. Frogs from two populations are harvested regularly throughout the year, while those in the third are protected from harvest. We compared seasonal and sex differences in diet, reproduction, and growth across these populations to examine life-history patterns. By volume, crayfish and anurans were the most abundant prey items for all adults across sites. Frogs from drier sites consumed more crayfish than frogs from the wettest site. Anurans were abundant in the diet during the wet season, while crayfish and fish were abundant during the dry season. More frogs with empty stomachs were captured during the wet season than the dry season. Feeding, growth, and fat deposition were greatest during the dry season across all sites. Although females were found in all reproductive stages throughout the year, the highest percentage of females had mature ova during the late dry season and spent ovaries during the early wet season. Individual patterns of growth were similar across all sites and matched historical growth data from the 1950s. Differences in body size among sites were most likely attributable to differential mortality (i.e., harvest pressure, predation) rather than to differences in food access or growth. ?? 2007 by the American Society of Ichthyologists and Herpetologists.

  1. Helminth infracommunities of Rana vaillanti brocchi (Anura: Ranidae) in Los Tuxtlas, Veracruz, Mexico.

    PubMed

    Paredes-Calderón, Laura; León-Règagnon, Virginia; García-Prieto, Luis

    2004-08-01

    A total of 76 adult individuals of Rana vaillanti were collected in Laguna Escondida, Los Tuxtlas, Veracruz, Mexico, and their helminth infracommunity structure was determined. Among the 21 helminth taxa collected (10 digeneans, 8 nematodes, and 3 acanthocephalans), the digenean Langeronia macrocirra reached the highest prevalence (64.4%), mean abundance (6.6), and mean intensity (10.4), as well as the highest total number of individuals (499). Only 2 frogs were uninfected, the remainder harbored between 1 and 7 helminth species and 1-102 individuals; mean species richness and abundance were 3.49 +/- 0.22 and 16.1 +/- 16.3, respectively. Langeronia macrocirra dominated in 50.6% of the infracommunities, with relatively low Berger-Parker index values (0.56); for this reason, the evenness was high (0.70 +/- 0.31), and consequently, diversity values are the highest recorded to date in species of Rana. However, patterns of helminth infracommunity richness and diversity were similar to those previously observed in amphibians. This structure is attributed to the feeding habits (between 66.7 and 81% of helminth species parasitizing R. vaillanti enter using the food web dynamics) and low vagility (the remainder species infect by host penetration).

  2. Highly complex mitochondrial DNA genealogy in an endemic Japanese subterranean breeding brown frog Rana tagoi (Amphibia, Anura, Ranidae).

    PubMed

    Eto, Koshiro; Matsui, Masafumi; Sugahara, Takahiro; Tanaka-Ueno, Tomoko

    2012-10-01

    The endemic Japanese frog Rana tagoi is unique among Holarctic brown frogs in that it breeds in small subterranean streams. Using mitochondrial 16S ribosomal RNA and NADH dehydrogenase subunit 1 genes, we investigated genealogical relationships among geographic samples of this species together with its relative R. sakuraii, which is also a unique stream breeder. These two species together form a monophyletic group, within which both are reciprocally paraphyletic. Rana tagoi is divided into two major clades (Clade A and B) that are composed of 14 genetic groups. Rana sakuraii is included in Clade A and split into two genetic groups, one of which forms a clade (Subclade A-2) with sympatric R. tagoi. This species-level paraphyly appears to be caused by incomplete taxonomy, in addition to introgressive hybridization and/or incomplete lineage sorting. Rana tagoi strongly differs from other Japanese anurans in its geographic pattern of genetic differentiation, most probably in relation to its unique reproductive habits. Taxonomically, R. tagoi surely includes many cryptic species.

  3. Behavioural adaptations of Rana temporaria to cold climates.

    PubMed

    Ludwig, Gerda; Sinsch, Ulrich; Pelster, Bernd

    2015-01-01

    Environmental conditions at the edge of a species' ecological optimum can exert great ecological or evolutionary pressure at local populations. For ectotherms like amphibians temperature is one of the most important abiotic factors of their environment as it influences directly their metabolism and sets limits to their distribution. Amphibians have evolved three ways to cope with sub-zero temperatures: freeze tolerance, freeze protection, freeze avoidance. The aim of this study was to assess which strategy common frogs at mid and high elevation use to survive and thrive in cold climates. In particular we (1) tested for the presence of physiological freeze protection, (2) evaluated autumnal activity and overwintering behaviour with respect to freeze avoidance and (3) assessed the importance of different high-elevation microhabitats for behavioural thermoregulation. Common frogs did not exhibit any signs of freeze protection when experiencing temperatures around 0 °C. Instead they retreated to open water for protection and overwintering. High elevation common frogs remained active for around the same period of time than their conspecifics at lower elevation. Our results suggest that at mid and high elevation common frogs use freeze avoidance alone to survive temperatures below 0 °C. The availability of warm microhabitats, such as rock or pasture, provides high elevation frogs with the opportunity of behavioural thermoregulation and thus allows them to remain active at temperatures at which common frogs at lower elevation cease activity.

  4. Terrestrial activity and conservation of adult California red-legged frogs Rana aurora draytonii in coastal forests and grasslands

    USGS Publications Warehouse

    Bulger, J.B.; Scott, N.J.; Seymour, R.B.

    2003-01-01

    The federally threatened California red-legged frog Rana aurora draytonii occupies both aquatic and terrestrial habitats in its adult life stage. The terrestrial activities of this species are not well known and require documentation to assist in the development of appropriate levels of protection under the US Endangered Species Act. We studied the terrestrial activities of radio-tagged red-legged frogs (n = 8-26) inhabiting a coastal watershed in Santa Cruz County, California, during 1997-1998. In particular, we investigated (1) the use of terrestrial habitats by non-migrating adults in relation to season, breeding chronology, and precipitation, and (2) adult migration behavior, including seasonal timing, duration, distances traveled, and the use of corridors. Non-migrating red-legged frogs occupied terrestrial habitats briefly (median = 4-6 days) following infrequent summer rains, but resided nearly continuously on land (median = 20-30 days) from the onset of the winter wet-season until breeding activities commenced 1-2 months later. All of the non-migrating frogs remained within 130 m of their aquatic site of residence (median <25 m). Intervals spent on land were again brief during mid/late winter (median = 1-4 days), despite frequent and copious rainfall. Adult migration to and from breeding sites occurred from late October through mid-May (wet season). We monitored 25 migration events between aquatic sites that were 200-2800 m apart. Short distance movements ( <300 m) were completed in 1-3 days, longer movements required up to 2 months. Most migrating frogs moved overland in approximately straight lines to target sites without apparent regard to vegetation type or topography. Riparian corridors were neither essential nor preferred as migration routes. Frogs traveling overland occurred in upland habitats as far as 500 m from water. Approximately 11-22% of the adult population was estimated to migrate to and from breeding sites annually, whereas the bulk of the

  5. Host-seeking heights, host-seeking activity patterns, and West Nile virus infection rates for members of the Culex pipiens complex at different habitat types within the hybrid zone, Shelby County, TN, 2002 (Diptera: Culicidae).

    PubMed

    Savage, Harry M; Anderson, Michael; Gordon, Emily; McMillen, Larry; Colton, Leah; Delorey, Mark; Sutherland, Genevieve; Aspen, Stephen; Charnetzky, Dawn; Burkhalter, Kristen; Godsey, Marvin

    2008-03-01

    Host-seeking heights, host-seeking activity patterns, and West Nile virus (family Flaviviridae, genus Flavivirus, WNV) infection rates were assessed for members of the Culex pipiens complex from July to December 2002, by using chicken-baited can traps (CT) at four ecologically different sites in Shelby County, TN. Host-seeking height was assessed by CT placed at elevations of 3.1, 4.6, and 7.6 m during one 24-h period per month. Host-seeking activity was assessed by paired CT placed at an elevation of 4.6 m. Can traps were sampled at one 10-h daytime interval and at seven 2-h intervals during the evening, night, and morning. Cx. pipiens complex mosquitoes accounted for 87.1% of collected mosquitoes. Culex (Melanoconion) erraticus (Dyar & Knab) accounted for 11.9% of specimens. The average number of Cx. pipiens complex mosquitoes collected per 24-h CT period from July to September was lowest at a rural middle income site (1.7), intermediate at an urban middle income site (11.3), and highest at an urban low income site (47.4). Can traps at the forested site failed to collect Cx. pipiens complex mosquitoes. From July to September at urban sites, Culex pipiens pipiens L. was the rarest of the three complex members accounting for 11.1-25.6% of specimens. At the rural site, Culex pipiens quinquefasciatus Say was the rarest member of the complex. Cx. p. pipiens was not collected after September. Mean abundance of Cx. pipiens complex mosquitoes was higher in traps at 7.6 m than in traps at 4.6 m. Abundances at 3.1 m were intermediate and not significantly different from abundances at the other heights. Initiation of host-seeking activity was associated with the end of civil twilight and activity occurred over an extended nighttime period lasting 8-10 h. All 11 WNV-positive mosquitoes were Cx. pipiens complex mosquitoes collected from urban sites in traps placed at elevations of 4.6 and 7.6 m. Infection rates were marginally nonsignificant by height. Infection rates, host

  6. A sex-linked Ace gene, not linked to insensitive acetylcholinesterase-mediated insecticide resistance in Culex pipiens.

    PubMed

    Malcolm, C A; Bourguet, D; Ascolillo, A; Rooker, S J; Garvey, C F; Hall, L M; Pasteur, N; Raymond, M

    1998-05-01

    An acetylcholinesterase (AChE) gene, Ace.x, showing 93% identity of deduced amino acid sequence to Anopheles stephensi Ace has been cloned from a Culex pipiens strain homozygous for insensitive AChE (iAChE) mediated insecticide resistance. DNA sequence of genomic DNA clones identified exons 2-5. RFLP of six clones indicated four possible alleles. Linkage analysis located Ace.x to chromosome I, less than 0.8 centimorgans from the sex locus, whereas the locus conferring resistance was 2.0 centimorgans from plum-eye on chromosome II. Ace.1 coding for AChE1, which is associated with resistance, is therefore autosomal. We propose that Ace.x is the recently postulated Ace.2 coding for the biochemically distinct AChE2, which is not associated with resistance.

  7. Effect of incubation at overwintering temperatures on the replication of West Nile Virus in New York Culex pipiens (Diptera: Culicidae).

    PubMed

    Dohm, D J; Turell, M J

    2001-05-01

    We examined the effect of simulated overwintering temperatures on West Nile (WN) virus replication in Culex pipiens L. derived from mosquitoes collected during the autumn 1999 WN epizootic in New York. The WN virus was a strain isolated from a dead crow also collected during this outbreak. Virus was recovered from most mosquitoes held exclusively at 26 degres C. In contrast, none of the mosquitoes held exclusively at the lower temperatures had detectable infections. When mosquitoes were transferred to 26 degrees C after being held at 10 degrees C for 21-42 d, infection and dissemination rates increased with increased incubation at 26 degrees C. Future studies involving the attempted isolation of WN virus from overwintering mosquitoes may benefit from holding the mosquitoes at 26 degrees C before testing for infectious virus.

  8. Effects of host species and life stage on the helminth communities of sympatric northern leopard frogs (Lithobates pipiens) and wood frogs (Lithobates sylvaticus) in the Sheyenne National Grasslands, North Dakota.

    PubMed

    Gustafson, Kyle D; Newman, Robert A; Tkach, Vasyl V

    2013-08-01

    We studied helminth communities in sympatric populations of leopard frogs (Lithobates pipiens) and wood frogs (Lithobates sylvaticus) and assessed the effects of host species and life stage on helminth community composition and helminth species richness. We examined 328 amphibians including 218 northern leopard frogs and 110 wood frogs collected between April and August of 2009 and 2010 in the Sheyenne National Grasslands of southeastern North Dakota. Echinostomatid metacercariae were the most common helminths found, with the highest prevalence in metamorphic wood frogs. Host species significantly influenced helminth community composition, and host life stage significantly influenced the component community composition of leopard frogs. In these sympatric populations, leopard frogs were common hosts for adult trematodes whereas wood frogs exhibited a higher prevalence of nematodes with direct life cycles. Metamorphic frogs were commonly infected with echinostomatid metacercariae and other larval trematodes whereas juvenile and adult frogs were most-frequently infected with directly transmitted nematodes and trophically transmitted trematodes. Accordingly, helminth species richness increased with the developmental life stage of the host.

  9. Larvicidal activity of selected plant hydrodistillate extracts against the house mosquito, Culex pipiens, a West Nile virus vector.

    PubMed

    Cetin, Huseyin; Yanikoglu, Atila; Cilek, James E

    2011-04-01

    The larvicidal activity of hydrodistillate extracts from Chrysanthemum coronarium L., Hypericum scabrum L., Pistacia terebinthus L. subsp. palaestina (Boiss.) Engler, and Vitex agnus castus L. was investigated against the West Nile vector, Culex pipiens L. (Diptera: Culicidae). Yield and identification of the major essential oils from each distillation was determined by GC-MS analyses. The major essential oil component for each plant species was as follows: α-pinene for P. terebinthus palaestina, and H. scabrum (45.3% and 42.3%, respectively), trans-β-caryophyllene for V. agnus castus (22.1%), and borneol for C. coronarium (20.9%). A series of distillate concentrations from these plants (that ranged from 1 ppm to 500 ppm, depending on plant species) were assessed against late third to early fourth C. pipiens larvae at 1, 6, and 24 h posttreatment. In general, larval mortality to water treated with a distillate increased as concentration and exposure time increased. H. scabrum and P. terebinthus palaestina were most effective against the mosquito larvae and both produced 100% mortality at 250 ppm at 24-h continuous exposure compared with the other plant species. Larval toxicity of the distillates at 24 h (LC(50) from most toxic to less toxic) was as follows: P. terebinthus palaestina (59.2 ppm) > H. scabrum (82.2 ppm) > V. agnus castus (83.3 ppm) > C. coronarium (311.2 ppm). But when LC(90) values were compared, relative toxicity ranking changed as follows: H. scabrum (185.9 ppm) > V. agnus castus (220.7 ppm) > P. terebinthus palaestina (260.7 ppm) > C. coronarium (496.3 ppm). Extracts of native Turkish plants continue to provide a wealth of potential sources for biologically active agents that may be applied against arthropod pests of man and animals.

  10. Airborne Pesticides as an Unlikely Cause for Population Declines of Alpine Frogs in the Sierra Nevada, California

    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...

  11. Predation by Oregon spotted frogs (Rana pretiosa) on Western toads (Bufo boreas) in Oregon, USA

    USGS Publications Warehouse

    Pearl, Christopher A.; Hayes, M.P.

    2002-01-01

    Toads of the genus Bufo co-occur with true frogs (family Ranidae) throughout their North American ranges. Yet, Bufo are rarely reported as prey for ranid frogs, perhaps due to dermal toxins that afford them protection from some predators. We report field observations from four different localities demonstrating that Oregon spotted frogs (Rana pretiosa) readily consume juvenile western toads (Bufo boreas) at breeding sites in Oregon. Unpalatability thought to deter predators of selected taxa and feeding mode may not protect juvenile stages of western toads from adult Oregon spotted frogs. Activity of juvenile western toads can elicit ambush behavior by Oregon spotted frog adults. Our review of published literature suggests that regular consumption of toadlets sets Oregon spotted frogs apart from most North American ranid frogs. Importance of the trophic context of juvenile western toads as a seasonally important resource to Oregon spotted frogs needs critical investigation.

  12. Workplace safety in Bangladesh ready-made garment sector: 3 years after the Rana Plaza collapse.

    PubMed

    Barua, Uttama; Ansary, Mehedi Ahmed

    2016-11-18

    Workplace safety is one of the most important issues in industries worldwide, and is endangered by industrial accidents. Different industrial disasters have resulted in several initiatives worldwide to protect human life and reduce material damage, both nationally and internationally. In Bangladesh, the ready-made garment (RMG) industry is one of the most important export-oriented business sectors, which is facing challenges to ensure workplace safety. The Rana Plaza collapse in Bangladesh is the consequence of such non-compliance. The accident resulted in different local and global initiatives to address the challenges. This article reviews progress and achievement of the initiatives to reduce vulnerability in the Bangladesh RMG industry within 3 years after the deadly accident. In the long run, the challenge is to maintain momentum already created for achieving sustainability in the RMG sector in Bangladesh and maintaining compliance even after the end of support from external partners.

  13. Hemodynamic consequences of delayed ventriculoconal conduction in the frog Rana catesbeiana.

    PubMed

    Liberthson, R R; Szidon, J P; Bharati, S; Lev, M; Fishman, A P

    1975-10-01

    We investigated the function of the conus arteriosus in the bullfrog Rana catesbeiana using a combination of anatomical and physiological techniques. Although there is a normal delay in ventriculoconal conduction and we could induce a spectrum of ventriculoconal conduction disturbances by manipulating the region of the ventriculoconal junction, we found no histological evidence of specialized conducting myocardial tissue in this region. The performance of the conus arteriosus was explored during various disturbances of ventriculoconal conduction and also during hemodynamic disturbances produced by hemorrhage and afterloading. The conus was found to contribute little to forward flow under ordinary circumstances, but its contribution increased greatly during bleeding or partial occlusion of the truncus. In contrast to the conclusion of others, no evidence could be adduced to support the idea that the conus serves as a depulsating chamber. Disparities in previous reports concerning the operation of the conus as a booster pump are attributed to special experimental circumstances.

  14. Fat body involvement in vitellogenin fate in the green frog, Rana esculenta.

    PubMed

    Varriale, B; Di Matteo, L; Minucci, S; Pierantoni, R; Chieffi, G

    1988-01-01

    1. Since, in Rana esculenta, fat bodies contain vitellogenin, the present study was performed in order to determine whether or not fat bodies are involved in the fate of vitellogenin. 2. The experiment of November shows that fat body excision provokes plasma vitellogenin increase even in animals treated with estradion-17 beta + pituitary crude homogenate (as compared with relative control). The same picture has been shown in the April experiment. 3. The result on protein-bound phosphate in ovaries from the April experiment has shown that fat body extirpation causes a decrease of protein-bound phosphate in the ovary. 4. This results indicates that fat bodies play an important role in sequestrating circulating vitellogenin by the ovary.

  15. The isolated and perfused working heart of the frog, Rana esculenta: an improved preparation.

    PubMed

    Acierno, R; Gattuso, A; Cerra, M C; Pellegrino, D; Agnisola, C; Tota, B

    1994-05-01

    1. An in vitro preparation of the intact heart of the frog Rana esculenta was set up. 2. The isolated heart, perfused at constant pressure, was spontaneously beating and able to generate physiological values of output pressure, cardiac output, ventricle work and power. It showed the typical phenomenon of the "hypodynamic state" after a relatively constant time from the onset of the perfusion. 3. Perfusion with air-saturated saline and 99.5% oxygen-saturated saline did not show significant differences in the recorded parameters. 4. This experimental model represents a useful tool for physiological and pharmacological studies, especially when the direct analysis of the effects of hormones, mediators or drugs requires an intact heart preparation.

  16. UBPy/MSJ-1 system during male germ cell progression in the frog, Rana esculenta.

    PubMed

    Meccariello, Rosaria; Chianese, Rosanna; Scarpa, Donatella; Berruti, Giovanna; Cobellis, Gilda; Pierantoni, Riccardo; Fasano, Silvia

    2007-01-01

    mUBPy (mouse ubiquitin specific processing protease) is a de-ubiquitinating enzyme expressed in mouse testis and brain. In testis, it interacts with the DnaJ protein MSJ-1 (mouse sperm cell specific DnaJ first homologue), a molecular chaperone expressed in spermatids and spermatozoa. Since MSJ-1 is conserved among vertebrates, to demonstrate an evolutionarily conserved function of UBPy/MSJ-1 system, we assayed mUBPy presence in the anuran amphibian, the frog, Rana esculenta, during the annual sexual cycle. By Western blot we have detected a specific signal of 126kDa in testis and isolated spermatozoa. During the annual sexual cycle, the signal gradually increases as soon as spermatogenesis resumes after the winter stasis. Using immunocytochemistry, we have localized the protein in spermatids and spermatozoa. In conclusion, UBPy/MSJ-1 system is available in R. esculenta testis suggesting a conserved fundamental function in spermatogenesis and sperm formation.

  17. Peptidomics and genomics analysis of novel antimicrobial peptides from the frog, Rana nigrovittata.

    PubMed

    Ma, Yufang; Liu, Cunbao; Liu, Xiuhong; Wu, Jing; Yang, Hailong; Wang, Yipeng; Li, Jianxu; Yu, Haining; Lai, Ren

    2010-01-01

    Much attention has been paid on amphibian peptides for their wide-ranging pharmacological properties, clinical potential, and gene-encoded origin. More than 300 antimicrobial peptides (AMPs) from amphibians have been studied. Peptidomics and genomics analysis combined with functional test including microorganism killing, histamine-releasing, and mast cell degranulation was used to investigate antimicrobial peptide diversity. Thirty-four novel AMPs from skin secretions of Rana nigrovittata were identified in current work, and they belong to 9 families, including 6 novel families. Other three families are classified into rugosin, gaegurin, and temporin family of amphibian AMP, respectively. These AMPs share highly conserved preproregions including signal peptides and spacer acidic peptides, while greatly diversified on mature peptides structures. In this work, peptidomics combined with genomics analysis was confirmed to be an effective way to identify amphibian AMPs, especially novel families. Some AMPs reported here will provide leading molecules for designing novel antimicrobial agents.

  18. Evaluation of the skin peptide defenses of the Oregon spotted frog Rana pretiosa against infection by the chytrid fungus Batrachochytrium dendrobatidis.

    PubMed

    Conlon, J Michael; Reinert, Laura K; Mechkarska, Milena; Prajeep, Manju; Meetani, Mohammed A; Coquet, Laurent; Jouenne, Thierry; Hayes, Marc P; Padgett-Flohr, Gretchen; Rollins-Smith, Louise A

    2013-06-01

    Population declines due to amphibian chytridiomycosis among selected species of ranid frogs from western North America have been severe, but there is evidence that the Oregon spotted frog, Rana pretiosa Baird and Girard, 1853, displays resistance to the disease. Norepinephrine-stimulated skin secretions were collected from a non-declining population of R. pretiosa that had been exposed to the causative agent Batrachochytrium dendrobatidis. Peptidomic analysis led to identification and isolation, in pure form, of a total of 18 host-defense peptides that were characterized structurally. Brevinin-1PRa, -1PRb, -1PRc, and -1PRd, esculentin-2PRa and -PRb, ranatuerin-2PRa, -2PRb, -2PRc, and -2PRe, temporin-PRb and -PRc were identified in an earlier study of skin secretions of frogs from a different population of R. pretiosa known to be declining. Ranatuerin-2PRf, -2PRg, -2PRh, temporin-PRd, -PRe, and -PRf were not identified in skin secretions from frogs from the declining population, whereas temporin-PRa and ranatuerin-2PRd, present in skin secretions from the declining population, were not detected in the current study. All purified peptides inhibited the growth of B. dendrobatidis zoospores. Peptides of the brevinin-1 and esculentin-2 families displayed the highest potency (minimum inhibitory concentration = 6.25-12.5 μM). The study provides support for the hypothesis that the multiplicity and diversity of the antimicrobial peptide repertoire in R. pretiosa and the high growth-inhibitory potency of certain peptides against B. dendrobatidis are important in conferring a measure of resistance to fatal chytridiomycosis.

  19. Comparative microhabitat characteristics at oviposition sites of the California red-legged frog (Rana draytonii)

    USGS Publications Warehouse

    Alvarez, Jeff A.; Cook, David G.; Yee, Julie L.; van Hattem, Michael G.; Fong, Darren R.; Fisher, Robert N.

    2013-01-01

    We studied the microhabitat characteristics of 747 egg masses of the federally-threatened Rana draytonii (California red-legged frog) at eight sites in California. our study showed that a broad range of aquatic habitats are utilized by ovipositing R. draytonii, including sites with perennial and ephemeral water sources, natural and constructed wetlands, lentic and lotic hydrology, and sites surrounded by protected lands and nested within modified urban areas. We recorded 45 different egg mass attachment types, although the use of only a few types was common at each site. These attachment types ranged from branches and roots of riparian trees, emergent and submergent wetland vegetation, flooded upland grassland/ruderal vegetation, and debris. eggs were deposited in relatively shallow water (mean 39.7 cm) when compared to maximum site depths. We found that most frogs in artificial pond, natural creek, and artificial channel habitats deposited egg masses within one meter of the shore, while egg masses in a seasonal marsh averaged 27.3 m from the shore due to extensive emergent vegetation. Rana draytonii appeared to delay breeding in lotic habitats and in more inland sites compared to lentic habitats and coastal sites. eggs occurred as early as mid-december at a coastal artificial pond and as late as mid-April in an inland natural creek. We speculate that this delay in breeding may represent a method of avoiding high-flow events and/or freezing temperatures. Understanding the factors related to the reproductive needs of this species can contribute to creating, managing, or preserving appropriate habitat, and promoting species recovery.

  20. The effect of shade on the container index and pupal productivity of the mosquitoes Aedes aegypti and Culex pipiens breeding in artificial containers.

    PubMed

    Vezzani, D; Albicócco, A P

    2009-03-01

    The aim of this study was to assess whether certain attributes of larval breeding sites are correlated with pupal productivity (i.e. numbers of pupae collected per sampling period), so that these could be used as the focus for control measures to enhance control efficiency. Therefore, the objectives were to identify the months of highest pupal productivity of Aedes aegypti (L.) and Culex pipiens L. (Diptera: Culicidae) in an urban temperate cemetery in Argentina where artificial containers of < 6 L (flower vases) were the predominant breeding habitats, to compare various measures of the productivity of sunlit and shaded containers and to determine whether the composition of the containers affected pupal productivity. Over a period of 9 months, 200 randomly chosen water-filled containers (100 sunlit and 100 shaded), out of approximately 3738 containers present (approximately 54% in shade), were examined each month within a cemetery (5 ha) in Buenos Aires (October 2006 to June 2007). In total, 3440 immatures of Cx pipiens and 1974 of Ae. aegypti were collected. The larvae : pupae ratio was 10 times greater for the former, indicating that larval mortality was greater for Cx pipiens. Both mosquito species showed a higher container index (CI) in shaded than in sunlit containers (Ae. aegypti: 12.8% vs. 6.9% [chi(2) = 17.6, P < 0.001]; Cx pipiens: 6.3% vs. 1.8% [chi(2) = 24, P < 0.001]). However, the number and the density of immatures per infested container and the number of pupae per pupa-positive container did not differ significantly between sunlit and shaded containers for either species. Therefore, the overall relative productivity of pupae per ha of Ae. aegypti and Cx pipiens was 2.3 and 1.8 times greater, respectively, in shaded than in sunlit areas as a result of the greater CIs of containers in shaded areas. Neither the CI nor the number of immatures per infested container differed significantly among container types of different materials in either lighting

  1. A Hierarchical Approach Embedding Hydrologic and Population Modeling for a West Nile Virus Vector Prediction

    NASA Astrophysics Data System (ADS)

    Jian, Y.; Silvestri, S.; Marani, M.; Saltarin, A.; Chillemi, G.

    2012-12-01

    We applied a hierarchical state space model to predict the abundance of Cx.pipiens (a West Nile Virus vector) in the Po River Delta Region, Northeastern Italy. The study area has large mosquito abundance, due to a favorable environment and climate as well as dense human population. Mosquito data were collected on a weekly basis at more than 20 sites from May to September in 2010 and 2011. Cx.pipiens was the dominant species in our samples, accounting for about 90% of the more than 300,000 total captures. The hydrological component of the model accounted for evapotranspiration, infiltration and deep percolation to infer, in a 0D context, the local dynamics of soil moisture as a direct exogenous forcing of mosquito dynamics. The population model had a Gompertz structure, which included exogenous meteorological forcings and delayed internal dynamics. The models were coupled within a hierarchical statistical structure to overcome the relatively short length of the samples by exploiting the large number of concurrent observations available. The results indicated that Cx.pipiens abundance had significant density dependence at 1 week lag, which approximately matched its development time from larvae to adult. Among the exogenous controls, temperature, daylight hours, and soil moisture explained most of the dynamics. Longer daylight hours and lower soil moisture values resulted in higher abundance. The negative correlation of soil moisture and mosquito population can be explained with the abundance of water in the region (e.g. due to irrigation) and the preference for eutrophic habitats by Cx.pipien. Variations among sites were explained by land use factors as represented by distance to the nearest rice field and NDVI values: the carrying capacity decreased with increased distance to the nearest rice filed, while the maximum growth rate was positively related with NDVI. The model shows a satisfactory performance in predicting (potentially one week in advance) mosquito

  2. Culex pipiens s.l. and Culex torrentium (Culicidae) in Wrocław area (Poland): occurrence and breeding site preferences of mosquito vectors.

    PubMed

    Weitzel, Thomas; Jawień, Piotr; Rydzanicz, Katarzyna; Lonc, Elzbieta; Becker, Norbert

    2015-01-01

    Both ornithophilic mosquito species, Culex pipiens s.l. (L.) and Culex torrentium (Martini, 1925), occur sympatric in temperate Europe. They are presumed to be primary vectors of West Nile and Sindbis viruses. Differentiation of these morphologically similar Culex species is essential for evaluation of different vector roles, for mosquito surveillance and integrated control strategies. Cx. torrentium has been neglected or erroneously determined as Cx. pipiens s.l. in some previous studies, because only males of both species can be diagnosed reliably by morphology. Thus, knowledge about species abundance, geographical distribution, breeding site preferences and the zoonotic risk assessment is incomplete also in Poland. In Wrocław area (Silesian Lowland), besides typical urban breeding sites, huge sewage irrigation fields provide suitable breeding conditions for Culex species. They are also inhabited by 180 resident and migratory bird species serving as potential virus reservoirs. In this study, morphology of larvae and males as well as species diagnostic enzyme markers, namely adenylate kinase (AK) and 2-hydroxybutyrate dehydrogenase (HBDH), were used to discriminate Cx. pipiens s.l. and Cx. torrentium. In a total of 650 Culex larvae from 24 natural and artificial breeding sites, Cx. pipiens s.l. had a proportion of 94.0% and Cx. torrentium only 6.0%. It could be shown that both species are well adapted to various breeding site types like ditches, catch basins, flower pots and buckets with diverse water quality. Cx. torrentium preferred more artificial water containers in urban surrounding (12% species proportion), whereas in semi-natural breeding sites, Cx. torrentium was rare (3%). In 12 of 24 breeding sites, larvae of both species have been found associated.

  3. Effects of chronic aluminum and copper exposure on growth and development of wood frog (Rana sylvatica) larvae.

    PubMed

    Peles, John D

    2013-09-15

    Wood frogs (Rana sylvatica) were exposed to aluminum (Al; 10, 100, 500, 1000, or 2000 μgL(-1)) or copper (Cu; 1, 10, 50, 100, 200 μgL(-1)) at a pH of 4.70 from the beginning of the larval period through the completion of metamorphosis (range=43-102 days). Observations on mortality, malformation, time to reach specific developmental stages, body mass at these stages, and metamorphic success were made throughout the larval developmental period. Only one case of malformation was observed and mortality was ≤ 10% at all concentrations except the highest Cu concentration where the rate was 33%. All larvae that survived the experiment successfully completed metamorphosis, but significant effects on growth and development occurred for both metals and these were most prominent for Cu. At the highest Al concentration (2000 μgL(-1)), body mass of larvae was significantly lower (reduced by 17% compared to the control) at 20 days post hatching (DPH) and the time to reach the hind-limb (HL), front-limb (FL), and tail resorption (TR) stages was significantly increased (9-10 days longer than the control). Body mass of larvae exposed to the three highest concentrations of Cu (50, 100, 200 μgL(-1)) was reduced by 30-34% at 20 DPH. Exposure to these concentrations also resulted in increased time to reach the HL, FL, and TR stages with larvae in the highest concentration taking 21-29 days longer to reach these stages. Larvae exposed to 10 μgL(-1) Cu also took longer to reach the FL and TR stages of development, and exposure to all Cu concentrations increased tail resorption time by more than two days compared to the control. Although the only observed effects of Al were for a concentration that is probably not ecologically relevant, results demonstrate that environmentally-realistic levels of Cu may have significant biological effects that could influence individual fitness and population-level processes.

  4. Effects of α-cypermethrin enantiomers on the growth, biochemical parameters and bioaccumulation in Rana nigromaculata tadpoles of the anuran amphibians.

    PubMed

    Xu, Peng; Huang, Ledan

    2017-02-13

    Populations of many amphibian species are declining worldwide in part because of pesticide contamination. As a surface water contaminant, α-cypermethrin may have severe ecological impacts on amphibians. Here, we examined the acute toxicity of α-cypermethrin enantiomers to dark-spotted frog Rana nigromaculata tadpoles at 24, 48, 72 and 96h, finding that the tadpoles were indeed sensitive to α-cypermethrin. The (S)-(1R, 3R)-enantiomer was approximately 29 times more toxic than the (R)-(1S, 3S)-enantiomer at 96h. A significant delayed growth in R. nigromaculata tadpoles after exposure to 0.5µgL(-1) of S-(1R, 3R)-cypermethrin was observed. Additionally, increased superoxide dismutase (SOD), catalase (CAT), glutathione-S-transferase (GST) and malondialdehyde (MDA) levels indicate the presence of oxidative stress in the tadpoles. Further, tadpoles exposed to sublethal concentrations of α-cypermethrin enantiomers exhibited enantioselective growth and oxidative damage. Bioaccumulation experiments showed that the tadpoles could rapidly accumulate α-cypermethrin. The (R)-(1S, 3S)-enantiomer was preferentially accumulated over the (S)-(1R, 3R)-enantiomer, and it was also eliminated more quickly, as evidenced in the subsequent depuration experiments.

  5. River islands, refugia and genetic structuring in the endemic brown frog Rana kukunoris (Anura, Ranidae) of the Qinghai-Tibetan Plateau.

    PubMed

    Zhou, Weiwei; Yan, Fang; Fu, Jinzhong; Wu, Shifang; Murphy, Robert W; Che, Jing; Zhang, Yaping

    2013-01-01

    Frequently, Pleistocene climatic cycling has been found to be the diver of genetic structuring in populations, even in areas that did not have continental ice sheets, such as on the Qinghai-Tibetan Plateau (QTP). Typically, species distributed on the plateau have been hypothesized to re-treat to south-eastern refugia, especially during the Last Glacial Maximum (LGM). We evaluated sequence variation in the mitochondrial DNA gene Cytb and the nuclear DNA gene RAG-1 in Rana kukunoris, a species endemic to the QTP. Two major lineages, N and S, were identified, and lineage N was further subdivided into N1 and N2. The geographical distribution and genealogical divergences supported the hypothesis of multiple refugia. However, major lineages and sublineages diverged prior to the LGM. Demographical expansion was detected only in lineage S and sublineage N2. Sublineage N1 might have survived several glacial cycles in situ and did not expand after the LGM because of the absence of suitable habitat; it survived in river islands. Genetic analysis and environment modelling suggested that the north-eastern edge of QTP contained a major refugium for R. kukunoris. From here, lineage S dispersed southwards after the LGM. Two microrefugia in northern Qilian Mountains greatly contributed to current level of intraspecific genetic diversity. These results were found to have important implications for the habitat conservation in Northwest China.

  6. Observations of Interspecific amplexus between western North American ranid frogs and the introduced American bullfrog (Rana catesbeiana) and an hypothesis concerning breeding interference

    USGS Publications Warehouse

    Pearl, Christopher A.; Hayes, M.P.; Haycock, Russ; Engler, Joseph D.; Bowerman, Jay

    2005-01-01

    Introduced American bullfrogs (Rana catesbeiana) come in contact with native amphibians on four continents and are well established in lowlands of western North America. To date, research on the effects of introduced bullfrogs on native frogs has focused on competition and predation, and is based largely on larval interactions. We present observations of interspecific amplexus between bullfrogs and two native ranid frogs (R. aurora and R. pretiosa) from six sites across the Pacific Northwest that imply that this interaction is more widespread than currently recognized. Our observations indicate that R. catesbeiana juveniles and subadults in this region are of appropriate size to elicit marked amplectic responses from males of both native species. Our literature review suggests that greater opportunity may exist for pairings between R. catesbeiana and native R. aurora or R. pretiosa than among syntopic native ranids in western North America. We hypothesize that interspecific amplexus with introduced R. catesbeiana could result in reproductive interference with negative demographic consequences in native ranid populations that have been reduced or altered by other stressors.

  7. [Role of acetylcholine in the Ca2+-dependent regulation of functional activity of myocardium of frog Rana temporaria].

    PubMed

    Shemarova, I V; Kuznetsov, S V; Demina, I N; Nesterov, V P

    2008-01-01

    To study role of ACh in the Ca2+-dependent regulation of rhythm and strength of cardiac contractions in frog Rana temporaria, the ACh chrono- and inotropic effects have been studied in parallel experiments on the background of blockers of potential-controlled Ca2+-channels, ryanodine and muscarine receptors. The obtained results indicate participation of acetylcholine in the Ca2+-dependent regulation of rhythm and strength of frog cardiac contractions.

  8. Preliminary Evidence that American Bullfrogs (Rana catesbeiana) Are Suitable Hosts for Escherichia coli O157:H7▿

    PubMed Central

    Gray, Matthew J.; Rajeev, Sreekumari; Miller, Debra L.; Schmutzer, A. Chandler; Burton, Elizabeth C.; Rogers, Emily D.; Hickling, Graham J.

    2007-01-01

    We orally inoculated Rana catesbeiana tadpoles (n = 23) and metamorphs (n = 24) to test their suitability as hosts for Escherichia coli O157:H7. Tadpoles were housed in flowthrough aquaria and did not become infected. Metamorphs were housed in stagnant aquaria, and 54% tested positive through 14 days postinoculation, suggesting that they are suitable hosts for E. coli O157:H7. PMID:17449685

  9. Bioaccumulation of macro- and trace elements by European frogbit (Hydrocharis morsus-ranae L.) in relation to environmental pollution.

    PubMed

    Polechońska, Ludmiła; Samecka-Cymerman, Aleksandra

    2016-02-01

    The aim of present study was to investigate the level of trace metals and macroelements in Hydrocharis morsus-ranae collected from regions differing in the degree and type of pollution. Concentrations of 17 macro- and microelements were determined in roots and shoots of European frogbit as well as in water and bottom sediments from 30 study sites. Plants differed in concentrations of elements and bioaccumulation capacity depending on the characteristics of dominant anthropogenic activities in the vicinity of the sampling site. Shoots of H. morsus-ranae growing in the vicinity of organic chemistry plants and automotive industry contained particularly high levels of Cd, Co, and S. Plants from area close to heat and power plant, former ferrochrome industry and new highway, were distinguished by the highest concentrations of Cr, Cu, and Pb. European frogbit from both these regions contained more Fe, Hg, Mn, Ni, and Zn than plants from agricultural and recreational areas. The concentrations of alkali metals and Co, Fe, and N in H. morsus-ranae were elevated in relation to the natural content in macrophytes irrespectively to their content in the environment. Based on the values of Bioaccumulation and Translocation Factors, European frogbit is an accumulator for Co, Cr, Cu, Fe, K, Mn, Ni, Pb, and Zn and a good candidate for phytoremediation of water polluted with Co, Cu, Hg, K, Mn, and Ni. The amount of Co and Mn removed from water and accumulated in the plant biomass during the vegetation season was considerably high.

  10. Diet of introduced bullfrogs (Rana catesbeiana): Predation on and diet overlap with native frogs on Daishan Island, China

    USGS Publications Warehouse

    Wu, Zhengjun; Li, Y.; Wang, Y.; Adams, Michael J.

    2005-01-01

    We examined diet of introduced Bullfrogs (Rana catesbeiana) and three native frog species (Rana limnocharis, Rana nigromaculata, and Bufo bufo gargarizans) co-occurring at a group of ponds on Daishan Island, east of China, to gain insight into the nature of potential interactions between Bullfrogs and native frog species. For postmetamorphic Bullfrogs, aquatic prey items dominated volumetrically. Prey size, diet volume and volumetric percentage of native frogs in diet increased with Bullfrog body size. The number and volumetric percentage of native frogs in the diet were not different for female and male Bullfrogs, and both were higher for adults than for juveniles. Diet overlap between males and juveniles was higher than that between males and females and between females and juveniles. Diet overlap with each native frog species of male Bullfrogs was lower than that of female Bullfrogs and juvenile Bullfrogs. We did not exam effects of Bullfrogs on native frogs but our results suggest that the primary threat posed by juvenile Bullfrogs to native frogs on Daishan Island is competition for food, whereas the primary threat posed by male Bullfrogs is direct predation. Female Bullfrogs may threaten native frogs by both competition and predation. These differences among Bullfrog groups may be attributed to differences in body size and microhabitat use.

  11. Cloning, Characterization, and Expression Analysis of MyD88 in Rana dybowskii.

    PubMed

    Niu, Shudong; Shi, Xuecan; Zhang, Jingyu; Chai, Longhui; Xiao, Xianghong

    2016-05-01

    The myeloid differentiation factor 88 (MyD88) is the most common adaptor protein in toll-like receptor (TLR) signaling pathways and plays an important role in the innate immune system. In this report, we conducted rapid amplification of complementary DNA (cDNA) ends (RACE), multiple sequence alignment, conserved domain search, phylogenetic tree construction, and quantitative real-time PCR to obtain and analyze the full-length cDNA sequence, the amino acid sequential structures, and the expression patterns of Rana dybowskii (Rd) MyD88. The full-length cDNA of RdMyD88 is 1472 bp, with an open reading frame of 855 bp, encoding a protein of 285 amino acid residues. The RdMyD88 amino acid sequence contains a death domain (DD) and a Toll/interleukin-1 receptor (TIR) domain. RdMyD88 was calculated as a hydrophilic protein with predicted molecular mass and pI of 32.79 kDa and 6.00, respectively. Eighteen possible phosphorylation sites including eight serine residues, six tyrosine residues, and four threonine residues are predicted. Analysis of multiple sequence alignment and phylogenetic tree revealed that the predicted RdMyD88 protein is closest to its Xenopus counterparts. The PCR result showed that RdMyD88 is expressed in various tissues of R. dybowskii. Quantitative real-time PCR (qPCR) was used to examine the expression of RdMyD88 in the heart, liver, and kidney. After Rana grylio virus (RGV) exposure, the expression of RdMyD88 in the heart, liver, and kidney were significantly upregulated and reached peak levels at 48, 48, and 72 h post-infection (hpi), respectively. Meanwhile, in response to Aeromonas hydrophila (AH) infection, clear upregulation of RdMyD88 was observed in the heart, liver, and kidney and reached its peak at 48, 6, and 12 hpi, respectively. The highest levels of induction were found in the kidney after both RGV and AH infections. These findings indicate that RdMyD88 has a conserved structure and is probably an important component of the innate

  12. Characterization of a new Aedes albopictus (Diptera: Culicidae)-Wolbachia pipientis (Rickettsiales: Rickettsiaceae) symbiotic association generated by artificial transfer of the wPip strain from Culex pipiens (Diptera: Culicidae).

    PubMed

    Calvitti, Maurizio; Moretti, Riccardo; Lampazzi, Elena; Bellini, Romeo; Dobson, Stephen L

    2010-03-01

    Wolbachia is a maternally inherited endosymbiont inducing various effects in insects and other invertebrate hosts that facilitate the invasion of naive host populations. One of the effects is a form of sterility known as cytoplasmic incompatibility (CI) through which females are effectively sterilized when they mate with males harboring a different Wolbachia strain. The repeated mass release of cytoplasmically incompatible males can be a tool to suppress insect populations. Here, we attempt to infect an Aedes albopictus (Skuse) (Diptera: Culicidae) strain, artificially deprived of the natural Wolbachia infection, with a new Wolbachia strain from Culex pipiens (L.) (Diptera: Culicidae). Further experiments were designed to study the effects of the new infection on Ae. albopictus fitness and evaluate key parameters that affect infection dynamics, including CI level and maternal inheritance. Using embryonic microinjection, the new Wolbachia strain was successfully established in Ae. albopictus. Crosses demonstrated a pattern of bidirectional CI between naturally infected and transinfected individuals. Specifically, egg hatch was essentially absent (i.e., CI was very high) in all crosses between the transinfected males and females with a different infection status. Furthermore, naturally infected Ae. albopictus males were incompatible with the transinfected females. Maternal inheritance was close to 100%. Moreover, the new infection did not affect immature and adult survivorship, but it significantly reduced female fecundity and egg hatch rate. The results are discussed in relation to the potential use of the new Ae. albopictus-Wolbachia symbiotic association as a suitable system for the study and development of CI-based strategies for suppressing populations of this important pest and disease vector.

  13. REPELLENT EFFECT OF OCIMUM BASILICUM AND GLYCYRRHIZA GLABRA EXTRACTS AGAINST THE MOSQUITO VECTOR, CULEX PIPIENS (DIPTERA: CULICIDAE).

    PubMed

    Hassan, Mostafa I; Hammad, Kotb M; Saeed, Saeed M

    2015-08-01

    Essential or volatile oils of plants have been variously reported to have many medicinal applications. Methanol, acetone and petroleum ether extracts of Ocimum basilicum and Glycyrrhiza glabra were screened for their repellency effect against Culex pipiens mosquito. The repellent action of the present plants extracts were varied depending on the solvent used and dose of extract. Methanol extract of O. basilicum exhibited the lowest repellent activity as it recorded 77.4% at 6.7mg/cm2. The petroleum ether and acetone extract of 0. basilicum showed repellency of 98.1 & 84.6% respectively, at dose of 6.7mg/cm2, while methanolic extract of G. glabra recorded 73.8 & 50.3% at dose of 6.7 &1.7mg/cm2 respectively, the petroleum ether and acetone extract of G. glabra showed repellency of 76.3 & 81.6%, respectively at dose of 6.7mg/cm2, compared with the commercial formulation, N.N. diethyl toulamide (DEET) which exhibited 100% repellent action at dose of 1.8mg/cm2, respectively. The results 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 meaure against mosquito bites and thus to control diseases caused by mosquito-borne pathogens.

  14. The diapause program impacts renal excretion and molecular expression of aquaporins in the northern house mosquito, Culex pipiens.

    PubMed

    Yang, Liu; Denlinger, David L; Piermarini, Peter M

    2016-12-27

    Adult females of the mosquito Culex pipiens entering diapause increase sugar water ingestion and reduce evaporative water loss, but how these attributes of the diapause program impact activity of the renal excretory system remains unknown. Here we compared the renal excretory capacity of diapausing and non-diapausing females, as well as the molecular expression of aquaporin (AQP) genes that encode channels involved in transporting water and/or small metabolites. Baseline urine excretion rates in diapausing mosquitoes were higher than in those of their non-diapausing counterparts, possibly a consequence of the intense sugar feeding associated with diapause. But, diapausing mosquitoes exhibited a much lower capacity for diuresis than non-diapausing mosquitoes. The suppressed diuretic capacity likely reflects reduced investment in the energetically-expensive post-prandial diuresis, an event not observed in diapausing mosquitoes. The mRNA expression levels of two genes encoding AQPs, Eglp1 and Aqp12L, in diapausing mosquitoes were down-regulated (on day 14) and up-regulated (on both days 3 and 14), respectively, in whole body samples. These changes were not evident in the excretory system (i.e., Malpighian tubules and hindgut), which showed no differential expression of AQPs as a function of diapause. Several AQP mRNAs were, however, differentially expressed in the midgut, ovaries, and abdominal body wall of diapausing mosquitoes, suggesting that AQPs in these tissues may be playing important non-excretory roles that are unique to diapause physiology.

  15. The role of miR-2∼13∼71 cluster in resistance to deltamethrin in Culex pipiens pallens.

    PubMed

    Guo, Qin; Huang, Yun; Zou, Feifei; Liu, Bingqian; Tian, Mengmeng; Ye, Wenyun; Guo, Juxin; Sun, Xueli; Zhou, Dan; Sun, Yan; Ma, Lei; Shen, Bo; Zhu, Changliang

    2017-03-22

    Excessive and continuous application of deltamethrin has resulted in the development of deltamethrin resistance among mosquitoes, which becomes a major obstacle for mosquito control. In a previous study, differentially expressed miRNAs between deltamethrin-susceptible (DS) strain and deltamethrin-resistant (DR) strain using illumina sequencing in Culex pipiens pallens were identified. In this study, we applied RNAi and the Centers for Disease Control and Prevention (CDC) bottle bioassay to investigate the relationship between miR-2∼13∼71 cluster (miR-2, miR-13 and miR-71) and deltamethrin resistance. We used quantitative real-time PCR (qRT-PCR) to measure expression levels of miR-2∼13∼71 clusters. MiR-2∼13∼71 cluster was down regulated in adult female mosquitoes from the DR strain and played important roles in deltamethrin resistance through regulating target genes, CYP9J35 and CYP325BG3. Knocking down CYP9J35 and CYP325BG3 resulted in decreased mortality of DR mosquitoes. This study provides the first evidence that miRNA clusters are associated with deltamethrin resistance in mosquitoes. Moreover, we investigated the regulatory networks formed between miR-2∼13∼71 cluster and its target genes, which provide a better understanding of the mechanism involved in deltamethrin resistance.

  16. Preoviposition activation of cathepsin-like proteinases in degenerating ovarian follicles of the mosquito Culex pipiens pallens.

    PubMed

    Uchida, K; Ohmori, D; Ueno, T; Nishizuka, M; Eshita, Y; Fukunaga, A; Kominami, E

    2001-09-01

    Within developing ovaries of many insects, some developing follicles or oocytes usually degenerate (follicular atresia or oosorption), while the others may continue to grow to maturity, thus maintaining the balance between the number of eggs and reproductive circumstances such as available nutrients. To help clarify the phenomenon of follicular atresia during ovarian development, we examined cysteine proteinases stored in mosquito Culex pipiens pallens ovaries. First, analysis using synthesized substrates showed that cathepsin B- and L-like proteinases gradually accumulated in the developing ovaries after a blood meal, which required more than 10 min of preincubation under acidic conditions to reach their maximum activities. However, homogenates of degenerating follicles 3 days after feeding showed proteolytic activities without acid treatment, suggesting that the proteinases had already been activated, while the extract of normally developing follicles collected from the same ovaries required more than 10 min of acid preincubation to reach the optimum activities, suggesting that the enzymes remained as inactive forms. Chemical and immunohistochemical analyses showed that more proteinases are located in the cytoplasm, rather than being associated with yolk granules. Ovarian proteinases, which are believed to become activated at the onset of embryogenesis, should also be activated during oogenesis, presumably to enhance oosorption.

  17. Evaluation of temephos and chlorpyrifos-methyl against Culex pipiens (Diptera: Culicidae) larvae in septic tanks in Antalya, Turkey.

    PubMed

    Cetin, H; Yanikoglu, A; Kocak, O; Cilek, J E

    2006-11-01

    The larvicidal activity of chlorpyrifos-methyl and temephos was evaluated against Culex pipiens L. (Diptera: Culicidae) in septic tanks in Antalya, Turkey. Chlorpyrifos-methyl (Pyrifos MT 25 emulsifiable concentrate [EC] ) was evaluated at application rates of 0.04, 0.08, and 0.12 mg active ingredient (AI)/liter, and temephos (Temeguard 50 EC) was evaluated at 0.02, 0.04, and 0.06 mg (AI)/liter during a 21-d study. Generally, overall larval reduction in septic tanks from single- and multifamily dwellings treated with either larvicide was significantly greater than pretreatment levels and control tanks for the duration of the study. At 14 d posttreatment, duration of control was greatest in multifamily tanks treated with chlorpyrifos-methyl at the highest application rate with similar levels of control through 21 d for single-family dwellings (range 97-100%). Septic tanks from both types of family dwellings treated at the highest application rate of temephos resulted in >90% reduction through day 21 (range 91-100%). Laboratory bioassays of septic tank water treated at field application rates, without daily dilution, revealed that complete larval mortality was achieved for 21 d at each application rate and formulation. It is thought that daily addition of water and organic matter to the septic tanks in the single and multifamily dwellings influenced the duration of effectiveness of the larvicides.

  18. Fecundity reduction in the second gonotrophic cycle of Culex pipiens infected with the apicomplexan blood parasite, Hepatozoon sipedon.

    PubMed

    Ferguson, Laura V; Smith, Todd G

    2014-08-01

    Fecundity reduction is a well-recognized phenomenon of parasite infection in insects. Reduced production of eggs might increase longevity of a host and release nutrients to both host and parasite that would otherwise be used for oogenesis. The objective of this study was to assess effects on fecundity caused by Hepatozoon sipedon, an apicomplexan blood parasite of snakes, in its invertebrate host, the mosquito Culex pipiens. In the first gonotrophic cycle, the mean number of eggs laid by mosquitoes infected with H. sipedon did not differ significantly from those laid by uninfected mosquitoes. However, in the second gonotrophic cycle infected mosquitoes laid significantly fewer eggs than did uninfected mosquitoes, and fecundity was reduced by 100% in mosquitoes with parasite burdens of more than 60 oocysts. There was a significant negative correlation between parasite burden, or the number of oocysts, and the number of eggs produced in the second gonotrophic cycle. Significantly fewer viable larvae hatched from eggs laid by infected compared to uninfected mosquitoes in the second gonotrophic cycle. These data indicate that fecundity reduction occurs in this system, although the physiological mechanisms driving this phenotype are not yet known.

  19. [Morpho-functional changes in small intestine epithelium of frog Rana temporaria during hibernation].

    PubMed

    Seliverstova, E V; Prutskova, N P

    2012-01-01

    Structure and function of small intestinal epithelium were studied in overwintering frogs Rana temporaria at various stages of hibernation. In the process of testing of absorption of arginine vasotocin (AVT) in experiments in vitro it is established that at the period of hibernation there is preserved the capability of the epithelium for absorption of this nonapeptide without hydrolysis. However, as compared with October-December, in January-February and later, a decrease of the AVT absorption takes place, which is the most pronounced in March-April. Changes in epithelial structures appear by the middle of winter and are progressing by spring. In April-May, as compared with the beginning of hibernation, the height of enterocytes, the length of microvilli, and the number of microvilli decrease by 33 %, 40 %, and 57 %, respectively. The absence of features of destruction indicates an adaptive character of the observed changes. Dynamics of the studied parameters indicates morphological plasticity of the small intestine epithelium of R. temporaria at the period of hibernation.

  20. Rana computatrix to human language: towards a computational neuroethology of language evolution.

    PubMed

    Arbib, Michael A

    2003-10-15

    Walter's Machina speculatrix inspired the name Rana computatrix for a family of models of visuomotor coordination in the frog, which contributed to the development of computational neuroethology. We offer here an 'evolutionary' perspective on models in the same tradition for rat, monkey and human. For rat, we show how the frog-like taxon affordance model provides a basis for the spatial navigation mechanisms that involve the hippocampus and other brain regions. For monkey, we recall two models of neural mechanisms for visuomotor coordination. The first, for saccades, shows how interactions between the parietal and frontal cortex augment superior colliculus seen as the homologue of frog tectum. The second, for grasping, continues the theme of parieto-frontal interactions, linking parietal affordances to motor schemas in premotor cortex. It further emphasizes the mirror system for grasping, in which neurons are active both when the monkey executes a specific grasp and when it observes a similar grasp executed by others. The model of human-brain mechanisms is based on the mirror-system hypothesis of the evolution of the language-ready brain, which sees the human Broca's area as an evolved extension of the mirror system for grasping.

  1. Effect of constant and fluctuating temperature on daily melatonin production by eyecups from Rana perezi.

    PubMed

    Valenciano, A I; Alonso-Gómez, A L; Alonso-Bedate, M; Delgado, M J

    1997-04-01

    We analysed the effect of daily temperature cycles in relation to constant temperature on day/night melatonin synthesis in frog eyecups in culture. Eyecups were cultured for 24 h under 12L:12D photoperiod and two thermal regimes, constant temperature (25, 15 and 5 degrees C) and thermoperiod (WL/CD, thermophase coinciding with photophase and cryophase coinciding with scotophase; and CL/WD, cryophase coinciding with photophase and thermophase coinciding with scotophase). A negative correlation between ocular serotonin N-acetyltransferase activity and culture temperature for both diurnal and nocturnal activities has been observed. This effect of increased ocular activity at low temperature is more pronounced than the well-known stimulatory effect of darkness, and it does not depend on the photoperiod phase. The lack of interactions between the phase of photoperiod and culture temperature indicates that the effects of both factors are independent. Night-time temperature is the key factor in determining the amplitude of the melatonin rhythm in the Rana perezi retina. However, daytime temperature can not counteract the inhibitory effect of light on ocular melatonin synthesis.

  2. Inhibitor and temperature effect on catalase in the liver of adult diploid and haploid Rana rugosa.

    PubMed

    Kashiwagi, A; Kashiwagi, K; Takase, M; Hanada, H; Yamashita, M; Naitoh, T; Nakamura, M

    1998-01-01

    The authors succeeded in raising a single mature haploid Rana rugosa female to the age of 2 years from an egg artificially fertilized with ultraviolet-irradiated sperm. In order to discover why this particular haploid individual should survive so long, hydrogen peroxide detoxifying catalase in the liver of this individual and age-matched diploids was examined and compared for total activity, temperature stability, and chemical inhibition. Total activity was found to be significantly higher in the haploid frog than in the diploids, suggesting that this particular haploid had a unique system for hydrogen peroxide detoxification which protected the liver against cell death, preventing hepatic failure, and leading to a prolonged survival. Liver catalase from the haploid proved to be more labile to aminotriazole and urea, losing 60-70% of its original activity after 30 min treatment, whereas diploid catalase lost only 40% under the same conditions. Haploid and diploid catalase responded similarly to heat, however. It seems likely that inhibitor-binding sites differ considerably between the catalase of normal diploids and the catalase of this particular haploid, while overall structure is generally similar.

  3. Physiological evidence for β3-adrenoceptor in frog (Rana esculenta) heart.

    PubMed

    Mazza, Rosa; Angelone, Tommaso; Pasqua, Teresa; Gattuso, Alfonsina

    2010-11-01

    β3-Adrenergic receptors (ARs) have been recently identified in mammalian hearts where, unlike β1- and β2-ARs, induce cardio-suppressive effects. The aim of this study was to describe β3-AR role in the frog (Rana esculenta) heart and to examine its signal transduction pathway. The presence of β3-AR, by using Western blotting analysis, has been also identified. BRL(37344), a selective β3-AR agonist, induced a dose-dependent negative inotropic effect at concentrations from 10(-12) to 10(-6)M. This effect was not modified by nadolol (β1/β2-AR antagonist) and by phentolamine (α-AR antagonist), but it was suppressed by the β3-AR-specific antagonist SR(59230) and by exposure to the Gi/o proteins inhibitor Pertussis Toxin. In addition, the involvement of EE-NOS-cGMP-PKG/PDE2 pathway in the negative inotropism of BRL(37344) has been assessed. BRL(37344) treatment induced eNOS and Akt phosphorylation as well as an increase of cGMP levels. β3-ARs activation induce a non-competitive antagonism against ISO stimulation which disappeared in presence of PKG and PDE2 inhibition. Taken together our findings provide, for the first time in the frog, a role for β3-ARs in the cardiac performance modulation which involves Gi/o protein and occurs via an EE-NO-cGMP-PKG/PDE2 cascade.

  4. Molecular cloning of natriuretic peptide receptor A from bullfrog (Rana catesbeiana) brain and its functional expression.

    PubMed

    Sekiguchi, T; Miyamoto, K; Mizutani, T; Yamada, K; Yazawa, T; Yoshino, M; Minegishi, T; Takei, Y; Kangawa, K; Minamino, N; Saito, Y; Kojima, M

    2001-08-08

    A comparative study of natriuretic peptide receptor (NPR) was performed by cloning the NPR-A receptor subtype from the bullfrog (Rana catesbeiana) brain and analyzing its functional expression. Like other mammalian NPR-A receptors, the bullfrog NPR-A receptor consists of an extracellular ligand binding domain, a hydrophobic transmembrane domain, a kinase-like domain and a guanylate cyclase domain. Sequence comparison among the bullfrog and mammalian receptors revealed a relatively low ( approximately 45%) similarity in the extracellular domain compared to a very high similarity ( approximately 92%) in the cytoplasmic regulatory and catalytic domains. Expression of NPR-A mRNA was detected in various bullfrog tissues including the brain, heart, lung, kidney and liver; highest levels were observed in lung. Functional expression of the receptor in COS-7 cells revealed that frog atrial natriuretic peptide (ANP) and brain natriuretic peptide (BNP) elicited cyclic guanosine 3'5'-monophosphate production by stimulating the receptor in a dose-dependent manner from 10(-10) M concentrations. Rat ANP was also effective in stimulating the frog receptor whereas rat BNP and porcine BNP were less responsive to the receptor. On the other hand, frog C-type natriuretic peptide (CNP) as well as porcine CNP stimulated the receptor only at high concentrations (10(-7) M). This clearly indicates that the bullfrog receptor is a counterpart of mammalian NPR-A, and is specific for ANP or BNP but not for CNP.

  5. A Threshold Dosage of Testosterone for Female-to-Male Sex Reversal in Rana rugosa Frogs.

    PubMed

    Oike, Akira; Kodama, Maho; Nakamura, Yoriko; Nakamura, Masahisa

    2016-10-01

    Androgens play a critical role in testicular differentiation in many species of vertebrates. While female-to-male sex reversal can be induced by testosterone (T) in some species of amphibians, the mechanism still remains largely unknown even at the histological level. In this study, we determined a threshold dosage of T to induce female-to-male sex reversal in the Japanese frog Rana (R.) rugosa. Tadpoles were allowed to metamorphose into frogs with T present in the rearing water. At 0.2 ng/mL T, female frogs formed tissue comprising a mixture of ovary and testis, the so-called ovotestis, the size of which was significantly smaller than the wild-type ovary. Histological changes occurring in the oocytes of T-treated ovaries induced oocyte degeneration in the masculinizing ovaries leading to their final disappearance. In parallel, many germ cells emerged in the cortex of the ovotestis and, later, in the medulla as well. RT-PCR analysis revealed upregulated expression of CYP17 and Dmrt1 but not 17βHSD in the ovotestis, and downregulation of Pat1a expression. Furthermore, immunohistology revealed CYP17-positive signals in the cortex of the masculinizing ovary, spreading throughout the whole area as the testis developed. These results indicate that oocytes are sensitive to T in the ovary of R. rugosa and that male-type germ cells expand in the masculinizing gonad (testis) contemporaneous with oocyte disappearance.

  6. Recent Emergence of a Chytrid Fungal Pathogen in California Cascades Frogs (Rana cascadae).

    PubMed

    De León, Marina E; Vredenburg, Vance T; Piovia-Scott, Jonah

    2016-12-12

    The pathogenic fungus Batrachochytrium dendrobatidis (Bd) has been associated with global amphibian declines, but it is often difficult to discern the relative importance of Bd as a causal agent in declines that have already occurred. Retrospective analyses of museum specimens have allowed researchers to associate the timing of Bd arrival with the timing of past amphibian declines. Cascades frogs (Rana cascadae) have experienced dramatic declines in northern California, but it is not clear whether the onset of these declines corresponds to the arrival of Bd. We used quantitative real-time PCR assays of samples collected from museum specimens to determine historical Bd prevalence in the northern California range of Cascades frogs. We detected Bd in 13 of 364 (3.5%) Cascades frog specimens collected between 1907 and 2003, with the first positive result from 1978. A Bayesian analysis suggested that Bd arrived in the region between 1973 and 1978, which corresponds well with the first observations of declines in the 1980s.

  7. Characterization and antioxidant activity in vitro and in vivo of polysaccharide purified from Rana chensinensis skin.

    PubMed

    Wang, Zhanyong; Zhao, Yuanyuan; Su, Tingting; Zhang, Jing; Wang, Fei

    2015-08-01

    Preliminary characterization and antioxidant activity in vitro and in vivo investigation of the polysaccharide fraction named as RCSP II, which was extracted from Rana chensinensis skin, were performed. Results indicated that RCSP II comprised glucose, galactose, and mannose in a molar ratio of 87.82:2.77:1.54 with a molecular weight of 12.8 kDa. Antioxidant activity assay in vitro showed that RCSP II exhibited 75.2% scavenging activity against 2,2'-azino-bis(3-ethylbenz-thiazoline-6-sulfonic acid) radicals at the concentration of 2500 mg/L and 85.1% against chelated ferrous ion at 4000 mg/L. Antioxidant activity assay in vivo further showed that RCSP II increased the activities of antioxidant enzymes, decreased the levels of malondialodehyde, and enhanced total antioxidant capabilities in livers and sera of d-galactose induced mice. These results suggested that RCSP II could have potential antioxidant applications as medicine or functional food.

  8. Differentiations of 5-HT and GAS cells in the digestive canals of Rana chensinensis tadpoles

    PubMed Central

    LI, Xin-Yi; LI, Qian; ZHANG, Yu-Hui

    2014-01-01

    In the current study, 5-nydroxytryptamine (5-HT) and gastrin (GAS) cells in the digestive canals of Rana chensinensis tadpoles at different developmental stages were investigated by immunohistochemistry. Results showed that the 5-HT cells were only detected in the duodenum before metamorphosis began, and were extensively distributed in the stomach, duodenum, small intestine, and rectum thereafter, with the highest counts found in the duodenum and rectum when metamorphosis was completed. The GAS cells were only distributed in the stomach and duodenum, and only rarely detected in the duodenum before metamorphosis began, but increased in the stomach during metamorphosis and showed zonal distribution in the gastric mucosa when metamorphosis was completed. Metamorphosis is a critical period for amphibians, during which structural and functional physiological adaptations are required to transition from aquatic to terrestrial environments. During metamorphosis, the differentiations of 5-HT cells in the gastrointestinal canals of tadpoles could facilitate mucus secretion regulation, improve digestive canal lubrication, and help watershortage food digestion in terrestrial environments. Conversely, GAS cell differentiations during metamorphosis might contribute to the digestive and absorptive function transition from herbivore to omnivore. PMID:25017753

  9. Growth and development of larval green frogs (Rana clamitans) exposed to multiple doses of an insecticide

    USGS Publications Warehouse

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

    2001-01-01

    Our objective was to determine how green frogs (Rana clamitans) are affected by multiple exposures to a sublethal level of the carbamate insecticide, carbaryl, in outdoor ponds. Tadpoles were added to 1,000-1 ponds at a low or high density which were exposed to carbaryl 0, 1, 2, or 3 times. Length of the larval period, mass, developmental stage, tadpole survival, and proportion metamorphosed were used to determine treatment effects. The frequency of dosing affected the proportion of green frogs that reached metamorphosis and the developmental stage of tadpoles. Generally, exposure to carbaryl increased rates of metamorphosis and development. The effect of the frequency of carbaryl exposure on development varied with the density treatment; the majority of metamorphs and the most developed tadpoles came from high-density ponds exposed to carbaryl 3 times. This interaction suggests that exposure to carbaryl later in the larval period stimulated metamorphosis, directly or indirectly, under high-density conditions. Our study indicates that exposure to a contaminant can lead to early initiation of metamorphosis and that natural biotic factors can mediate the effects of a contaminant in the environment.

  10. Effects of nonylphenol on rates of tail resorption and metamorphosis in Rana catesbeiana tadpoles.

    PubMed

    Christensen, Jennie R; Richardson, John S; Bishop, Christine A; Pauli, Bruce; Elliott, John

    2005-04-09

    Nonylphenol (NP) is a persistent, lipophilic, and toxic chemical that can be endocrine disrupting (estrogenic) at sublethal concentrations. Since amphibian metamorphosis is a hormone-driven process and a delicate balance of hormone levels is required for successful metamorphosis, exposure of larval amphibians to NP might disrupt metamorphic processes. This study tested whether NP exposure influenced rate of metamorphic progression and tail resorption in bullfrog (Rana catesbeiana) tadpoles. Premetamorphic bullfrog tadpoles were exposed for 7 d to one of 3 nominal concentrations of NP (234 microg/L, 468 microg/L, or 936 microg/L) with or without the addition of exogenous 3,3',5-triiodothyronine (T3). In the absence of exogenous T3, NP significantly increased the rate of tail growth (as measured by tail length) at 936 microg/L. There was no significant effect of NP alone on tail width, limb development, or the process of cranial transformation. When T3 was added to the treatments, increasing NP concentrations were associated with a significant decrease in the rate of cranial transformation, and at the highest dose, the rate of tail resorption was significantly lower than in the controls. Overall, NP had an inhibitory effect on the rate of bullfrog tadpole metamorphic progression and tail resorption.

  11. Effects of fluoride on development and growth of Rana chensinensis embryos and larvae.

    PubMed

    Chai, Lihong; Dong, Suiming; Zhao, Hongfeng; Deng, Hongzhang; Wang, Hongyuan

    2016-04-01

    The present study examined the adverse effects of fluoride exposure on embryos and larvae of Rana chensinensis. Survival, morphological abnormalities, growth and development, time to metamorphosis and size at metamorphic climax of R. chensinensis were examined. Our results showed that embryos malformation occurred in all fluoride treatments. Morphological abnormalities of embryos are characterized by axial flexures, the extrusion of fin axis, edema, and ruffled dorsal and ventral fin. Additionally, 4.1mg F(-)/L and above could significantly inhibit embryos growth and development. On day 15, total length and weight of tadpole were significantly lower in 19.6 and 42.4 mg F(-)/L treatments compared to control. However, significant reductions in total length and weight were observed only at 42.4 mg F(-)/L on day 30. Moreover, significant metamorphic delay and decrease in the size at metamorphic climax were found in larvae exposed to 42.4 mg F(-)/L. Taken together, embryos of R. chensinensis are more vulnerable to fluoride exposure than their tadpoles. Our results suggested that the presence of high concentrations fluoride might increase mortality risk and a reduction in juvenile recruitment in the field by increasing embryos malformation, delaying metamorphosis and decreasing size at metamorphosis.

  12. Mobile phone mast effects on common frog (Rana temporaria) tadpoles: the city turned into a laboratory.

    PubMed

    Balmori, Alfonso

    2010-06-01

    An experiment has been made exposing eggs and tadpoles of the common frog (Rana temporaria) to electromagnetic radiation from several mobile (cell) phone antennae located at a distance of 140 meters. The experiment lasted two months, from the egg phase until an advanced phase of tadpole prior to metamorphosis. Measurements of electric field intensity (radiofrequencies and microwaves) in V/m obtained with three different devices were 1.8 to 3.5 V/m. In the exposed group (n = 70), low coordination of movements, an asynchronous growth, resulting in both big and small tadpoles, and a high mortality (90%) was observed. Regarding the control group (n = 70) under the same conditions but inside a Faraday cage, the coordination of movements was normal, the development was synchronous, and a mortality of 4.2% was obtained. These results indicate that radiation emitted by phone masts in a real situation may affect the development and may cause an increase in mortality of exposed tadpoles. This research may have huge implications for the natural world, which is now exposed to high microwave radiation levels from a multitude of phone masts.

  13. Novel family of antimicrobial peptides from the skin of Rana shuchinae.

    PubMed

    Zheng, Ruiqiang; Yao, Bin; Yu, Haining; Wang, Hanjin; Bian, Jianmin; Feng, Feifei

    2010-09-01

    So far numerous antimicrobial peptides have been characterized from amphibians. In this work, a new family of antimicrobial peptides, named shuchin, was purified and characterized from skin secretions of the frog, Rana shuchinae that lives in freezing mountains. Totally two members of shuchin (shuchin 1 and 2) were identified with the amino acid sequence of NALSMPRNKCNRALMCFG and NALSSPRNKCDRASSCFG, respectively. cDNAs encoding shuchins were cloned from the skin cDNA library of R. shuchinae. The precursors of shuchin are composed of 62 amino acid residues including the conserved signal peptides, acidic propieces, and mature antimicrobial peptides. Synthetic shuchins showed strong and broad antimicrobial activities against Gram-positive bacteria (Staphylococcus aureus, and Bacillus cereus; MICs<12.5 microg/ml), Gram-negative bacteria (Escherichia coli, Bacillus dysenteriae, Pseudomonas aeruginosa; most MICs from 3.1 to 12.5 microg/ml), and yeast (Candida albicans; MICs of 6.25 microg/ml), but no hemolytic activity under the effective concentration, thereby provide more leading templates for designing novel anti-infection agents.

  14. Rana grylio Virus (RGV) 50L Is Associated with Viral Matrix and Exhibited Two Distribution Patterns

    PubMed Central

    Lei, Xiao-Ying; Ou, Tong; Zhang, Qi-Ya

    2012-01-01

    Background The complete genome of Rana grylio virus (RGV) was sequenced and analyzed recently, which revealed that RGV 50L had homologues in many iridoviruses with different identities; however, the characteristics and functions of 50L have not been studied yet. Methodology/Principal Findings We cloned and characterized RGV50L, and revealed 50L functions in virus assembly and gene regulation. 50L encoded a 499-amino acid structural protein of about 85 kDa in molecular weight and contained a nuclear localization signal (NLS) and a helix- extension-helix motif. Drug inhibition assay demonstrated that 50L was an immediate-early (IE) gene. Immuno-fluorescence assay revealed that 50L appeared early and persisted in RGV-infected cells following two distribution patterns. One pattern was that 50L exhibited a cytoplasm-nucleus- viromatrix distribution pattern, and mutagenesis of the NLS motif revealed that localization of 50L in the nucleus was NLS-dependent; the other was that 50L co-localized with viral matrix which plays important roles in virus assembly and the life circle of viruses. Conclusions/Significance RGV 50L is a novel iridovirus IE gene encoded structural protein which plays important roles in virus assembly. PMID:22912781

  15. Lead concentrations in bullfrog Rana catesbeiana and green frog R. clamitans tadpoles inhabiting highway drainages

    USGS Publications Warehouse

    Birdsall, C.W.; Grue, C.E.; Anderson, A.

    1986-01-01

    Lead concentrations were determined in sediment and tadpoles of bullfrogs Rana catesbeiana and green frogs R. clamitans from drainages along highways with different daily average traffic volumes (range, 4272 to I08,800 vehicles day-I) and from ponds >0.4 km from the nearest highway. Lead concentrations (mg kg--I dry weight) in sediment (7-8 to 940) were usually greater (4-5 times) than those in the tadpoles (bullfrog, 0,07 to 270; green frog, 0,90 to 240 mg kg-I). Lead concentrations in sediment (r =0.63) and in both species of tadpoles (bullfrog, r = 0.69; green frog, r = 0.57) were positively correlated with average daily traffic volume. Lead concentrations in both species of tadpoles (bullfrog, r = (). 76: green frog, r = 0.75) were also positively correlated with lead concentrations in sediment. At sites where both bullfrog and green frog tadpoles were collected. lead concentrations in the two species were closely related (r = 0.84). Lead concentrations in tadpoles living near highways may contribute to the elevated lead levels reported in wildlife that are potential tadpole predators. Dietary lead concentrations similar to those in our tadpoles have been associated with physiological and reproductive effects in some species of birds and mammals. However, additional data are needed to determine the hazards to predators of lead concentrations in tadpoles.

  16. Non-specific immune response of bullfrog Rana catesbeiana to intraperitoneal injection of bacterium Aeromonas hydrophila

    NASA Astrophysics Data System (ADS)

    Zhang, Junjie; Zou, Wenzheng; Yan, Qingpi

    2008-08-01

    Non-specific immune response of bullfrog Rana catesbeiana to pathogenic Aeromonas hydrophila was studied to 60 individuals in two groups. Each bullfrog in bacterium-injected group was injected intraperitoneally (i.p.) with 0.2 ml bacterial suspension at a density of 5.2 × 106 CFU/ml, while each one in control group injected i.p. with 0.2 ml sterile saline solution (0.85%, w/v). Three bullfrogs in both groups were sampled at 0, 1, 3, 7, 11, 15 and 20 days post-injection (dpi) for the evaluation of non-specific immune parameters. It was observed that intraperitoneal injection of A. hydrophila significantly increased the number of leucocytes and that of NBT-positive cells in peripheral blood. Significant increases in serum bactericidal activity and serum acid phosphatase activity were also observed in the bacterium-injected frogs when compared with those in the control group. However, a significant reduction was detected in vitro in phagocytosis activity of peripheral blood phagocytes. No significant difference in changes in the number of peripheral erythrocytes, serum superoxide dismutase (SOD) activity, and lysozyme activity was detected between the two groups. It is suggested that bullfrogs may produce a series of non-specific immune reactions in response to the A. hydrophila infection.

  17. [Reabsorption of yellow fluorescent protein in the Rana temporaria kidney by receptor-mediated endocytosis].

    PubMed

    Seliverstova, E V; Prutskova, N P

    2014-01-01

    The absorption of yellow fluorescent protein (YFP) and the expression of the endocytic receptors, megalin and cubilin, were investigated in the renal proximal tubules (PT) in frogs Rana temporaria after parenteral YFP injections. The methods of confocal microscopy and immunohistochemistry were used. The dynamics of YFP absorption was analyzed 2 h after injection. The logarithmic time dependence of the accumulation of YFP-containing endocytic vesicles in PT cells and the completion of absorption process 90-120 min after injection were shown. Unlike substantial megalin and cubilin expression 15-30 min after YFP introduction, immunolabeled endocytic receptors were not detected in PT cells after 2 h. The re-injection of YFP led to the appearance of apical endocytic vesicles containing megalin or cubilin colocalized with YFP. At the same time, the decrease of YFP uptake associated with reduction in the number of receptor-containing vesicles was demonstrated, suggesting a failure of megalin and cubilin expression. The decrease of absorption capacity of PT cells after YFP re-injection was similar to that found previously under conditions of the competitive absorption of green fluorescent protein (GFP) and YFP injected in different sequences. The data are the further demonstration of the proposed mechanism limiting the tubular protein absorption in the frog kidney and suggest the involvement of megalin and cubilin in uptake and vesicular transport of YFP.

  18. [Reinnervation of a mixed muscle in the frog Rana temporaria with a regenerating homogeneous nerve].

    PubMed

    Radziukevich, T L

    1995-01-01

    Mixed muscle m. iliofibularis from the frog Rana temporaria, consisting of monosynaptically innervated phasic and polysynaptically innervated postural muscle fibers, was reinnervated by homogeneous tailor's nerve having no axons of tonic motor system in its composition. Within 2-7 months after nerves binding treatment phasic muscle fibers were easily identified by subneural apparatus structure revealed at coloration of synaptic acetylcholinesterase. Presynaptic part of neuromuscular apparatus of these fibers after the impregnation by protargol was presented by immature nervous terminals. The identification of tonic Muscular fibers was difficult especially at the late stages of reinnervation as subneural apparatus structure typical for tonic fibers was not revealed. Nonmyelinizated nerve fibers without features of terminal branch were observed in individual regions of nonidentified muscle fibers. The results obtained show that subneural apparatus of tonic muscle fibers depends to a great extent on the influence of inherent tonic motor system. Axons of phasic motor system even at distant reinnervation periods and in the absence of competitive influences of tonic motor system do not form typical "phasic" terminal picture of innervation under the contact with tonic muscle fibers.

  19. Efficacy of potential chemical control compounds for removing invasive American bullfrogs (Rana catesbeiana).

    PubMed

    Witmer, Gary W; Snow, Nathan P; Moulton, Rachael S

    2015-01-01

    Invasive American bullfrogs [Rana catesbeiana (Lithobates catesbeianus)] are outcompeting and predating on native biota and contributing to reductions in biodiversity worldwide. Current methods for controlling American bullfrogs are incapable of stopping their expansion, thus more cost-effective and broadly applicable methods are needed. Although chemical control compounds have been identified as effective for removing other invasive amphibians, none have been tested for American bullfrogs. Our objective was to expand on previous research and test the efficacy of 10 potential chemical control compounds for removing invasive American bullfrogs. After a dermal spray-application of 4 ml, we found 3 compounds (i.e., chloroxylenol, rotenone with permethrin, and caffeine) at 5-10 % concentrations in water were 100 % lethal for adult American bullfrogs. Chloroxylenol and rotenone with permethrin were fast acting with time-to-death <2 h. This research presents a first-step toward incorporating chemical control as part of integrated pest management strategy for controlling invasive American bullfrogs. Follow-up studies on delivery systems and reducing non-target hazards should ensue with these compounds to confirm their effectiveness and safety for removing invasive American bullfrogs.

  20. Antimicrobial peptides with atypical structural features from the skin of the Japanese brown frog Rana japonica.

    PubMed

    Isaacson, Todd; Soto, AnaMaria; Iwamuro, Shawichi; Knoop, Floyd C; Conlon, J Michael

    2002-03-01

    Japonicin-1 (FFPIGVFCKIFKTC) and japonicin-2 (FGLPMLSILPKALCILLKRKC), two peptides with differential growth-inhibitory activity against the Gram-negative bacterium, Escherichia coli and the Gram-positive bacterium Staphylococcus aureus, were isolated from an extract of the skin of the Japanese brown frog Rana japonica. Both peptides show little amino acid sequence similarity to previously characterized antimicrobial peptides isolated from the skins of Ranid frogs. Circular dichroism studies, however, demonstrate that japonicin-2 adopts an alpha-helical conformation in 50% trifluoroethanol in common with many other cationic antimicrobial peptides synthesized in amphibian skin. Peptides belonging to the brevinin-1, brevinin-2, and tigerinin families, previously identified in the skins of Asian Ranid frogs, were not detected but a temporin-related peptide (ILPLVGNLLNDLL.NH(2); temporin-1Ja), that atypically bears no net positive charge, was isolated from the extract. The minimum inhibitory concentrations (MICs) of the peptides against E. coli were japonicin-1, 30 microM; japonicin-2, 12 microM; and temporin-1Ja > 100 microM. The MICs against S. aureus were japonicin-1, > 100 microM; japonicin-2, 20 microM; and temporin-1Ja, > 100 microM.

  1. Evidence for Directional Selection at a Novel Major Histocompatibility Class I Marker in Wild Common Frogs (Rana temporaria) Exposed to a Viral Pathogen (Ranavirus)

    PubMed Central

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

    2009-01-01

    Whilst the Major Histocompatibility Complex (MHC) is well characterized in the anuran Xenopus, this region has not previously been studied in another popular model species, the common frog (Rana temporaria). Nor, to date, have there been any studies of MHC in wild amphibian host-pathogen systems. We characterise an MHC class I locus in the common frog, and present primers to amplify both the whole region, and specifically the antigen binding region. As no more than two expressed haplotypes were found in over 400 clones from 66 individuals, it is likely that there is a single class I locus in this species. This finding is consistent with the single class I locus in Xenopus, but contrasts with the multiple loci identified in axolotls, providing evidence that the diversification of MHC class I into multiple loci likely occurred after the Caudata/Anura divergence (approximately 350 million years ago) but before the Ranidae/Pipidae divergence (approximately 230 mya). We use this locus to compare wild populations of common frogs that have been infected with a viral pathogen (Ranavirus) with those that have no history of infection. We demonstrate that certain MHC supertypes are associated with infection status (even after accounting for shared ancestry), and that the diseased populations have more similar supertype frequencies (lower FST) than the uninfected. These patterns were not seen in a suite of putatively neutral microsatellite loci. We interpret this pattern at the MHC locus to indicate that the disease has imposed selection for particular haplotypes, and hence that common frogs may be adapting to the presence of Ranavirus, which currently kills tens of thousands of amphibians in the UK each year. PMID:19240796

  2. Polypeptides from the Skin of Rana chensinensis Exert the Antioxidant and Antiapoptotic Activities on HaCaT Cells.

    PubMed

    Zhang, Xin; Cheng, Yunyun; Yang, Yang; Liu, Songcai; Shi, Hui; Lu, Chao; Li, Siming; Nie, Linyan; Su, Dan; Deng, Xuming; Ding, Kexiang; Hao, Linlin

    2017-01-02

    Studies have shown that frog skin secretes many types of peptides that are good for human skin. In this study, acid and enzymatic extracts of Rana skin peptides (acid/enzymatic Rana skin peptides, ARPs/ERPs) were obtained. The chemical and physical properties of the ARPs and ERPs were identified through UV scanning, HGLC, FRIT, and MS. MTS and flow cytometry were used to test the proproliferative and antiapoptotic effects of the ARPs and ERPs on human immortalized keratinocytes (HaCaT cells). To elucidate the antiapoptotic mechanisms, the mRNA and protein levels of EGF (epidermal growth factor, which enhances stimulation of cellular proliferation in both cells and epithelial tissues) and caspase-3 were evaluated using quantitative RT-PCR. The results indicated that the ARPs and ERPs were extracted from the Rana skin with yields of 0.65% and 0.52%, respectively. Treatment with ARPs (1.6 g/L) and ERPs (0.8 g/L) showed a 1.66-fold (p < 0.001) and 2.1-fold (p < 0.001) enhancement in the proliferation rates of HaCaT cells. The rate of apoptosis decreased by 2.6 fold (p < 0.01) and 3.4 fold (p < 0.01) under the UVB stimulation, respectively, at the same time, the up-regulation of EGF and down-regulation of caspase-3 were found. These results suggested that we can dig into the potential value of ARPs/ERPs in a new field.

  3. Total salivary gland proteins of female Culex pipiens and Aedes caspius (Diptera: Culicidae) and their fractionation during adult development and after blood sucking.

    PubMed

    Soliman, M A; Abdel-Hamid, M E; Mansour, M M; Seif, A I; Kamel, K I; el Hamshary, E M

    1999-08-01

    Salivary glands of Culex pipiens and Aedes caspius were analyzed to determine total protein content and its fractionation during adult female development and after blood sucking. In both species, the molecular weight of proteins ranged between 26.000 and 84.000 Daltons. These proteins were not identical in the two species. In Cx. pipiens, the total protein level increased during the first 3 days of adult development from 4.94 +/- 0.84 to 6.6 +/- 0.37 micrograms/gland. During this period, the salivary gland proteins were separated into 35, 34 and 37 fractions respectively. Cx. pipiens released in the human host 64% of the total proteins while taking a blood meal compared to unfed females. This decrease in protein level was proportional to protein fractions. Over the next 6 days, the protein level increased again to attain values comparable to those obtained prior to blood sucking. In Ae. caspius, the total protein level of the salivary glands did not change during the first 4 days of adult development (range between 3.13 +/- 0.27 and 3.91 +/- 0.36 micrograms gland), but on the fifth day, 2-fold increase was observed. The total salivary gland protein increased during the next 3 days after blood sucking to reach 15.5 +/- 0.98 micrograms/gland. During this period, a tremendous change in protein patterns was observed. After oviposition, on the fourth day, a significant reduction in the total protein level was observed (4.13 +/- 0.56 micrograms/gland), but over the next 3 days the level increased again (range between 4.13 +/- 0.66 and 7.13 +/- 0.66 micrograms/gland).

  4. Evaluation of halofenozide against prey mosquito larvae Culex pipiens and the predator fish Gambusia affinis: impact on growth and enzymatic activities.

    PubMed

    Soltani, N; Chouahda, S; Smagghe, G

    2008-01-01

    Culex pipiens (Diptera: Culicidae) is the most widely distributed mosquito species in Algeria and many other countries in the world. Mosquitoes are generally controlled by conventional insecticides but these may pose strong secondary effects on the environment. In this context, the insect growth regulators (IGRs) have shown promise in controlling pest insects. Halofenozide (23% EC) is a novel IGRs belonging to the class of non-steroidal ecdysone agonists, and it was found toxic for larvae of C. pipiens. In addition biological methods constitute an alternative to chemical control. Several fish species have been tested against mosquitoes, and Gambusia affinis was found very efficient. In the present study we evaluated the impact of this new potent insecticide (halofenozide) on growth and metric indexes in the larvivorous fish G. affinis under laboratory conditions. In addition, the effects were evaluated on the enzymatic activities of acetyl cholinesterase (AChE) and glutathione S-transferase (GST). The insecticide was added in water at two concentrations (12.6 and 28.6 microg/L) corresponding to the LC50 and LC90 obtained against fourth instar larvae of C. pipiens, and adult females of G. affinis were exposed to halofenozide for 30 days. At different exposure times we measured the length and weight of fishes, the index of condition (K), the gonado-somatic ratio (GSR) and the hepato-somatic ratio (HSR). The results showed that halofenozide had no significant (p>0.05) effects on growth, metric indexes and AChE activities. However, treatment caused a significant induction (p<0.05) in GST activities at days 15 and 30 with the highest dose. Our results indicate that this ecdysteroid agonist presented only minor secondary effects on the non-target fish species, and so it has potential for controlling of mosquitoes in an integrated manner.

  5. Operational Evaluation Of Vectomax® WSP (Bacillus thuringiensis Subsp. israelensis+Bacillus sphaericus) Against Larval Culex pipiens in Septic Tanks (1).

    PubMed

    Cetin, Huseyin; Oz, Emre; Yanikoglu, Atila; Cilek, James E

    2015-06-01

    The residual effectiveness of VectoMax® WSP (a water-soluble pouch formulation containing a combination of Bacillus thuringiensis subsp. israelensis strain AM65-52 and B. sphaericus strain ABTS 1743) when applied to septic tanks against 3rd- and 4th-stage larvae of Culex pipiens L. was evaluated in this study. This formulation was evaluated at operational application rates of 1 pouch (10 g) and 2 pouches (20 g) per septic tank. Both application rates resulted in >96% control of larvae for 24 days. Operationally, VectoMax WSP has proven to be a useful tool for the nonchemical control of Culex species in septic tank environments.

  6. The effects of laboratory Hepatozoon gracilis infection on the fecundity, mortality and longevity of Culex (Culex) pipiens Linneaus (Diptera: Culicidae) in Egypt.

    PubMed

    Adham, Fatma K; Gabre, Refaat M; Ayaad, Tahany H; Galal, Fatma H

    2003-08-01

    Laboratory observations on the effect of Hepatozoon gracilis on the egg production of the mosquito Cx. (Cx.) pipiens Linneaus under laboratory conditions revealed that H. gracilis infected mosquitoes produced significantly fewer eggs than uninfected ones. The egg production decreased as parasite burdens increased. Reduction in blood meal size in infected females did not reduce fecundity. No size differences was detected between oocyst-infected and uninfected females although sporozoite positive females were significantly large. Preoviposition period was affected significantly, while incubation period and percentage of egg hatching showed no significant changes. The longevity of female infected mosquitoes decreased insignificantly than in uninfected ones.

  7. Digestive enzyme as benchmark for insecticide resistance development in Culex pipiens larvae to chemical and bacteriologic insecticides.

    PubMed

    Kamel, Nashwa H; Bahgat, Iman M; El Kady, Gamal A

    2013-04-01

    This work monitored changes in some digestive enzymes (trypsin and aminopeptidase) associated with the building up of resistance in Cx. pipiens larvae to two chemical insecticides (methomyl and/or malathion) and one biological insecticide (Bacillus thuringiensis-H14 or B.t H 14). The LC50 value of methomyl for both field- and the 12th generation (F12) of the selected strain was 1.789 ppm and 8.925 ppm respectively. The LC50 value of malathion for both field and the F12 of the selected strain was 0.082 ppm and 0.156 ppm respectively, and those of B.t H14 of field strain and the F12 was 2.550ppm & 2.395ppm respectively. The specific activity of trypsin enzyme in control susceptible colony was 20.806 +/- 0.452micromol/min/mg protein; but at F4 and F8 for malathion and methomyl treated larvae were 10.810 +/- 0.860 & 15.616+/-0.408 micromol/min/mg protein, respectively. Trypsin activity of F12 in treated larvae with B.t.H14 was 2.097 +/- 0.587 microiol/min/mg protein. Aminopeptidase specific activity for susceptible control larvae was 173.05 +/- 1.3111 micromol/min/mg protein. This activity decreased to 145.15 +/- 4.12, 152.497 +/- 6.775 & 102.04 +/- 3.58a micromol/min/mg protein after larval (F 12) treatment with methomyl, malathion and B.t H 14 respectively.

  8. Structure and expression strategy of the genome of Culex pipiens densovirus, a mosquito densovirus with an ambisense organization.

    PubMed

    Baquerizo-Audiot, Elizabeth; Abd-Alla, Adly; Jousset, Françoise-Xavière; Cousserans, François; Tijssen, Peter; Bergoin, Max

    2009-07-01

    The genome of all densoviruses (DNVs) so far isolated from mosquitoes or mosquito cell lines consists of a 4-kb single-stranded DNA molecule with a monosense organization (genus Brevidensovirus, subfamily Densovirinae). We previously reported the isolation of a Culex pipiens DNV (CpDNV) that differs significantly from brevidensoviruses by (i) having a approximately 6-kb genome, (ii) lacking sequence homology, and (iii) lacking antigenic cross-reactivity with Brevidensovirus capsid polypeptides. We report here the sequence organization and transcription map of this virus. The cloned genome of CpDNV is 5,759 nucleotides (nt) long, and it possesses an inverted terminal repeat (ITR) of 285 nt and an ambisense organization of its genes. The nonstructural (NS) proteins NS-1, NS-2, and NS-3 are located in the 5' half of one strand and are organized into five open reading frames (ORFs) due to the split of both NS-1 and NS-2 into two ORFs. The ORF encoding capsid polypeptides is located in the 5' half of the complementary strand. The expression of NS proteins is controlled by two promoters, P7 and P17, driving the transcription of a 2.4-kb mRNA encoding NS-3 and of a 1.8-kb mRNA encoding NS-1 and NS-2, respectively. The two NS mRNAs species are spliced off a 53-nt sequence. Capsid proteins are translated from an unspliced 2.3-kb mRNA driven by the P88 promoter. CpDNV thus appears as a new type of mosquito DNV, and based on the overall organization and expression modalities of its genome, it may represent the prototype of a new genus of DNV.

  9. Female Choice for Males with Greater Fertilization Success in the Swedish Moor Frog, Rana arvalis

    PubMed Central

    Sherman, Craig D. H.; Sagvik, Jörgen; Olsson, Mats

    2010-01-01

    Background Studies of mate choice in anuran amphibians have shown female preference for a wide range of male traits despite females gaining no direct resources from males (i.e. non-resource based mating system). Nevertheless, theoretical and empirical studies have shown that females may still gain indirect genetic benefits from choosing males of higher genetic quality and thereby increase their reproductive success. Methodology/Principal Findings We investigated two components of sexual selection in the Moor frog (Rana arvalis), pre-copulatory female choice between two males of different size (‘large’ vs. ‘small’), and their fertilization success in sperm competition and in isolation. Females' showed no significant preference for male size (13 small and six large male preferences) but associated preferentially with the male that subsequently was the most successful at fertilizing her eggs in isolation. Siring success of males in competitive fertilizations was unrelated to genetic similarity with the female and we detected no effect of sperm viability on fertilization success. There was, however, a strong positive association between a male's innate fertilization ability with a female and his siring success in sperm competition. We also detected a strong negative effect of a male's thumb length on his competitive siring success. Conclusions/Significance Our results show that females show no preference for male size but are still able to choose males which have greater fertilization success. Genetic similarity and differences in the proportion of viable sperm within a males ejaculate do not appear to affect siring success. These results could be explained through pre- and/or postcopulatory choice for genetic benefits and suggest that females are able to perceive the genetic quality of males, possibly basing their choice on multiple phenotypic male traits. PMID:21049015

  10. Hormonal induction of spermatozoa from amphibians with Rana temporaria and Bufo bufo as anuran models.

    PubMed

    Uteshev, V K; Shishova, N V; Kaurova, S A; Browne, R K; Gakhova, E N

    2012-01-01

    The use of hormonally induced spermatozoa expressed in urine (HISu) is a valuable component of reproduction technologies for amphibians. Five protocols for sampling HISu from the European common frog (Rana temporaria) were compared: (1) pituitary extracts, (2) 0.12 µg g⁻¹ luteinising hormone-releasing hormone analogue (LHRHa), (3) 1.20 µg g⁻¹ LHRHa, (4) 11.7 IU g⁻¹ human chorionic gonadotrophin (hCG) and (5) 23.4 IU g⁻¹ hCG (g⁻¹ = per gram bodyweight). From 1 to 24h after administration we assessed the number and concentration of spermatozoa in spermic urine and in holding water, and in urine the percentage of motile spermatozoa and their progressive motility. The protocol using 1.20 µg g⁻¹ LHRHa gave the highest total sperm numbers (650 × 10⁶) and the highest percentage (40%) of samples with sperm concentrations above 200 × 10⁶ mL⁻¹. The percentage motility and progressive motility was similar from all protocols. Considerable amounts of spermatozoa were expressed by R. temporaria into their holding water. We tested hormonal priming and spermiation in the common toad (Bufo bufo) using 0.13 µg g⁻¹ LHRHa administered 24h before a final spermiating dose of 12.8 IU g⁻¹ hCG. No spermatozoa were expressed in holding water. Priming resulted in 35% more spermatozoa than without; however, there were no differences in sperm concentrations. Primed B. bufo produced spermatozoa with significantly higher percentage motility, but not progressive motility, membrane integrity, or abnormal spermatozoa than unprimed males.

  11. Metabolism of C26 bile alcohols in the bullfrog, Rana catesbeiana

    SciTech Connect

    Noma, Y.; Kihira, K.; Kuramoto, T.; Hoshita, T.

    1988-03-01

    Metabolism of C26 bile alcohols in the bullfrog, Rana catesbeiana, was studied. (24-14C)-24-Dehydro-26-deoxy-5 beta-ranol (3 alpha,7 alpha,12 alpha-trihydroxy-27-nor-5 beta-cholestan-24-one) was chemically synthesized from (24-14C)cholic acid and incubated with bullfrog liver homogenate fortified with NADPH. 24-Dehydro-26-deoxy-5 beta-ranol was shown to be converted into both 26-deoxy-5 beta-ranol and 24-epi-26-deoxy-5 beta-ranol ((24S)- and (24R)-27-nor-5 beta-cholestane-3 alpha,7 alpha,12 alpha,24-tetrols) in addition to 5 beta-ranol ((24R)-27-nor-5 beta-cholestane-3 alpha,7 alpha,12 alpha,24,26-pentol), which is the major bile alcohol of the bullfrog. (24-3H)-26-Deoxy-5 beta-ranol and (24-3H)-24-epi-26-deoxy-5 beta-ranol were prepared from 24-dehydro-26-deoxy-5 beta-ranol by reduction with sodium (3H) borohydride and administered respectively to two each of four bullfrogs by intraperitoneal injection. After 24 h, labeled 5 beta-ranol was isolated from the bile of the bullfrogs that received (24-3H)-26-deoxy-5 beta-ranol. In contrast little if any radioactivity could be detected in 5 beta-ranol or its 24-epimer after administration of (24-3H)-24-epi-26-deoxy-5 beta-ranol.

  12. Gonadal differentiation in frogs, Rana japonica and R. brevipoda, raised from UV irradiated eggs

    SciTech Connect

    Shirane, T.

    1982-10-10

    The gonadal differentiation of anurans, Rana japonica and R. brevipoda, was examined in animals raised from eggs which had been irradiated at the vegetal hemisphere with UV (9300 erg/mm2) at the 2-cell stage. In R. japonica about 70% of the larvae at stage I from the pressed and UV-irradiated eggs were germ cell free, but at a stage immediately after metamorphosis all animals had at least some germ cells, although their gonads often were extremely small and poorly differentiated. When male animals matured sexually, many of them had abnormal gonads. However, all of them were shown by artificial means to be capable of fertilization. In the nonpressed and irradiated group, no larvae were germ cell free and the animals immediately after metamorphosis showed nearly normal gonadal differentiation except for the presence of a few degenerate oocytes in the ovaries. The results in R. brevipoda were basically similar to those in R. japonica. In both species, sex ratios were determined at two stages, the first immediately after metamorphosis and the other when the animals matured, as based on gonad morphology and histology and on external sexually dimorphic characters as well. Sex ratios at these two stages in frogs from the pressed and irradiated eggs differed markedly in R. brevipoda. The ratio was normal at metamorphosis but high M/F ratios occurred when animals became mature. That sex reversal took place in this species as well as in R. japonica (in which sex-ratio deviation was not statistically significant) was supported by the sex ratios of the progenies of these supernumerary males.

  13. Motor planning modulates sensory-motor control of collision avoidance behavior in the bullfrog, Rana catesbeiana

    PubMed Central

    Nakagawa, Hideki; Nishida, Yuuya

    2012-01-01

    Summary In this study, we examined the collision avoidance behavior of the frog, Rana catesbeiana to an approaching object in the upper visual field. The angular velocity of the frog's escape turn showed a significant positive correlation with the turn angle (r2 = 0.5741, P<0.05). A similar mechanism of velocity control has been known in head movements of the owl and in human saccades. By analogy, this suggests that the frog planned its escape velocity in advance of executing the turn, to make the duration of the escape behavior relatively constant. For escape turns less than 60°, the positive correlation was very strong (r2 = 0.7097, P<0.05). Thus, the frog controlled the angular velocity of small escape turns very accurately and completed the behavior within a constant time. On the other hand, for escape turns greater than 60°, the same correlation was not significant (r2 = 0.065, P>0.05). Thus, the frog was not able to control the velocity of the large escape turns accurately and did not complete the behavior within a constant time. In the latter case, there was a small but significant positive correlation between the threshold angular size and the angular velocity (r2 = 0.1459, P<0.05). This suggests that the threshold is controlled to compensate for the insufficient escape velocity achieved during large turn angles, and could explain a significant negative correlation between the turn angle and the threshold angular size (r2 = 0.1145, P<0.05). Thus, it is likely that the threshold angular size is also controlled by the turn angle and is modulated by motor planning. PMID:23213389

  14. Toxic effects of endrin and toxaphene on the southern leopard frog Rana sphenocephala

    USGS Publications Warehouse

    Hall, R.J.; Swineford, D.

    1980-01-01

    Eggs, larvae and sub-adults of the southern leopard frog Rana sphenocephala were exposed to endrin and toxaphene. Exposure was in water by a continuous-flow technique, following standards that have been used successfully in the study of fish and invertebrates. R. sphenocephala is more sensitive to both pesticides than are higher vertebrates but is slightly less sensitive than fish. Eggs seem to be resistant to the effects of both pesticides and are probably poor indicators of environmental hazard. The toxic level of endrin is about equal in larvae and transformed frogs (LC50, 0?005-0?015 ppm). Toxaphene is less toxic to sub-adults (LC50, 0?37-0?790 ppm) than to larvae (LC50, 0?032-0?054 ppm). Delayed mortality, behavioural aberrations and effects on growth have been seen in toxaphene-dosed larvae observed over 30-day periods. Behavioural effects are more severe than those reported in other groups of animals. Effects on growth resulting from a 96-h exposure begin in the 0?013-0?018 ppm range. The maximum accumulation of residues observed for each chemical represented bioconcentration factors of about 100. Endrin residues are apparently lost more readily than toxaphene residues; relative depuration rates correlate well with the time course of toxic action in each chemical. Although less sensitive to these pesticides than fish, amphibians may not be protected in their natural habitats. Future studies of the effects of toxicants on amphibians should employ larvae if only one stage can be tested, should expose subjects for at least 96 h and should continue observations for a total of at least 30 days.

  15. The osteochondral ligament: a fibrous attachment between bone and articular cartilage in Rana catesbeiana.

    PubMed

    Felisbino, S L; Carvalho, H F

    2000-12-01

    The anuran epiphyseal cartilage shows a lateral expansion that covers the external surface of the bone, besides other features that distinguish it from the corresponding avian and mammalian structures. The fibrous structure that attaches the lateral cartilage to the bone was characterized in this work. It was designated osteochondral ligament (OCL) and presented two main areas. There was an inner area that was closer to the periosteal bone and contained a layer of osteoblasts and elongated cells aligned to and interspersed with thin collagen fibers. The thin processes of the cells in this area showed strong alkaline phosphatase activity. The outer area, which was closer to the cartilage, was rich in blood vessels and contained a few cells amongst thick collagen fibers. TRITC-phaloidin staining showed the cells of the inner area to be rich in F-actin, and were observed to form a net around the cell nucleus and to fill the cell processes which extended between the collagen fibers. Cells of the outer area were poor in actin cytoskeleton, while those associated with the blood vessels showed intense staining. Tubulin-staining was weak, regardless of the OCL region. The main fibers of the extracellular matrix in the OCL extended obliquely upwards from the cartilage to the bone. The collagen fibers inserted into the bone matrix as Sharpey's fibers and became progressively thicker as they made their way through the outer area to the cartilage. Immunocytochemistry showed the presence of type I and type III collagen. Microfibrils were found around the cells and amongst the collagen fibrils. These microfibrils were composed of either type VI collagen or fibrilin, as shown by immunocytochemistry. The results presented in this paper show that the osteochondral ligament of Rana catesbeiana is a complex and specialized fibrous attachment which guarantees a strong and flexible anchorage of the lateral articular cartilage to the periosteal bone shaft, besides playing a role in bone

  16. Molecular cloning and characterization of oocyte-specific Pat1a in Rana rugosa frogs.

    PubMed

    Nakamura, Yoriko; Iwasaki, Takehiro; Umei, Yosuke; Saotome, Kazuhiro; Nakajima, Yukiko; Kitahara, Shoichi; Uno, Yoshinobu; Matsuda, Yoichi; Oike, Akira; Kodama, Maho; Nakamura, Masahisa

    2015-10-01

    The Pat1 gene is expressed in the immature oocytes of Xenopus, and is reportedly involved in regulating the translation of maternal mRNAs required for oocyte-maturation. However, it is still unknown when Pat1a first appears in the differentiating ovary of amphibians. To address this issue, we isolated the full-length Pat1a cDNA from the frog Rana rugosa and examined its expression in the differentiating ovary of this frog. Among eight different tissues examined, the Pat1a mRNA was detectable in only the ovary. When frozen sections from the ovaries of tadpoles at various stages of development were immunostained for Vasa-a germ cell-specific protein-and Pat1a, Vasa-immunopositive signals were observed in all of the germ cells, whereas Pat1a signals were confined to the growing oocytes (50-200 μm in diameter), and absent from small germ cells (<50 μm in diameter). Forty days after testosterone injection into tadpoles to induce female-to-male sex-reversal, Pat1a-immunoreactive oocytes had disappeared completely from the sex-reversed gonad, but Vasa-positive small germ cells persisted. Thus, Pat1a would be a good marker for identifying the sexual status of the sex-reversing gonad in amphibians. In addition, fluorescence in situ hybridization analysis showed Pat1a to have an autosomal locus, suggesting that Pat1a transcription is probably regulated by a tissue-specific transcription factor in R. rugosa.

  17. Regulation of SMAD transcription factors during freezing in the freeze tolerant wood frog, Rana sylvatica.

    PubMed

    Aguilar, Oscar A; Hadj-Moussa, Hanane; Storey, Kenneth B

    2016-11-01

    The wood frog, Rana sylvatica, survives sub-zero winter temperatures by undergoing full body freezing for weeks at a time, during which it displays no measurable brain activity, no breathing, and a flat-lined heart. Freezing is a hypometabolic state characterized by a global suppression of gene expression that is elicited in part by transcription factors that coordinate the activation of vital pro-survival pathways. Smad transcription factors respond to TGF-β signalling and are involved in numerous cellular functions from development to stress. Given the identity of genes they regulate, we hypothesized that they may be involved in coordinating gene expression during freezing. Protein expression of Smad1/2/3/4/5 in response to freezing was examined in 24h frozen and 8h thawed wood frog tissues using western immunoblotting, with the determination of subcellular localization in muscle and liver tissues. Transcript levels of smad2, smad4 and downstream genes (serpine1, myostatin, and tsc22d3) were measured by RT-PCR. Tissue-specific responses were observed during freezing where brain, heart, and liver had elevated levels of pSmad3, and skeletal muscle and kidneys had increased levels of pSmad1/5 and pSmad2 during freeze/thaw cycle, while protein and transcript levels remained constant. There were increases in nuclear levels of pSmad2 in muscle and pSmad3 in liver. Transcript levels of serpine1 were induced in heart, muscle, and liver, myostatin in muscle, and tsc22d3 in heart, and liver during freezing. These results suggest a novel freeze-responsive activation of Smad proteins that may play an important role in coordinating pro-survival gene networks necessary for freeze tolerance.

  18. Effect of acidic precipitation on amphibian breeding in temporary ponds in Pennsylvania. [Rana sylvatica; Ambystoma jeffersonianum

    SciTech Connect

    Freda, J.; Dunson, W.A.

    1985-11-01

    This study assessed the impacts of acid deposition on amphibians breeding in temporary ponds in Pennsylvania by investigating the lowest pH's at which embryos could hatch, the physiological effects of low pH on amphibian larvae, pond chemistry and the influence of rainfall on pond pH, and the effect of pond pH on embryonic survival and local distribution of Ambystoma jeffersonianum and Rana sylvatica. At very low pH's, embryos stopped development soon after exposure. At higher but still lethal pH's, embryos became curled and failed to hatch. Embryos of Ambystoma were able to hatch even though they were curled, but R. sylvatica became trapped and died. Acute exposure to low pH's depressed sodium influx and accelerated sodium efflux, with a net loss of 50% of body sodium resulting in death. Increasing the external calcium concentration extended survival time by slowing the loss of sodium. Chronic exposure to low pH's resulted in reduction in body sodium, but to a lesser degree. R sylvatica tadpoles from a low pH pond had lower body sodium than tadpoles from a nearby high pH pond. Tadpoles from both ponds placed in a low pH pond underwent higher sodium efflux than when placed in the high pH pond. In studying the effect of low environmental pH, A. jeffersonianum was intolerant of low pH and was absent from most acidic ponds. R. sylvatica was tolerant and was found in ponds with the lowest pH. 73 refs., 14 figs., 21 tabs.

  19. D-Asp: a new player in reproductive endocrinology of the amphibian Rana esculenta.

    PubMed

    Raucci, Franca; Di Fiore, Maria Maddalena

    2011-11-01

    We investigated the involvement of D-Aspartic acid (D-Asp) on ovarian and testicular morphology of the green frog, Rana esculenta, and its effect on the testosterone production. The study has been performed throughout the reproductive cycle. In both ovary and testis a substantial amount of D-Asp is endogenously present and its concentration varies as function of reproduction. In the frog, D-Asp content is differently correlated with gonadal and plasmatic levels of testosterone, depending on the sex. In fact, the amount of the D-Asp is inversely linked with that of the testosterone in the ovary, while this correlation directly matched in the testis. In vivo short-term experiments, consisting of a single intra-peritoneal injection of D-Asp (2.0 μmol/g body weight), demonstrated that the enantiomer is significantly accumulated by both the ovary and testis, reaching after 3 h the highest uptake and thereafter decreasing to baseline values within 24 h. Furthermore, D-Asp influences the synthesis and/or the release of testosterone, causing a decrease of its level in the female, and an increase in the male, respectively. In vivo long-term experiments, D-Asp, chronically administered to the frogs of both sexes, enhances the maturation of both gonads, determining in the oocytes an higher accumulation of carbohydrate yolk plates in the ooplasm, and stimulating the spermatogenesis in the testis. Taken altogether, our results show that D-Asp operates differently in female and male frog gonads, indicating that it has different targets in the reproductive machinery depending on the sex.

  20. Motor planning modulates sensory-motor control of collision avoidance behavior in the bullfrog, Rana catesbeiana.

    PubMed

    Nakagawa, Hideki; Nishida, Yuuya

    2012-11-15

    In this study, we examined the collision avoidance behavior of the frog, Rana catesbeiana to an approaching object in the upper visual field. The angular velocity of the frog's escape turn showed a significant positive correlation with the turn angle (r(2) = 0.5741, P<0.05). A similar mechanism of velocity control has been known in head movements of the owl and in human saccades. By analogy, this suggests that the frog planned its escape velocity in advance of executing the turn, to make the duration of the escape behavior relatively constant. For escape turns less than 60°, the positive correlation was very strong (r(2) = 0.7097, P<0.05). Thus, the frog controlled the angular velocity of small escape turns very accurately and completed the behavior within a constant time. On the other hand, for escape turns greater than 60°, the same correlation was not significant (r(2) = 0.065, P>0.05). Thus, the frog was not able to control the velocity of the large escape turns accurately and did not complete the behavior within a constant time. In the latter case, there was a small but significant positive correlation between the threshold angular size and the angular velocity (r(2) = 0.1459, P<0.05). This suggests that the threshold is controlled to compensate for the insufficient escape velocity achieved during large turn angles, and could explain a significant negative correlation between the turn angle and the threshold angular size (r(2) = 0.1145, P<0.05). Thus, it is likely that the threshold angular size is also controlled by the turn angle and is modulated by motor planning.

  1. Gonadal differentiation in frogs, Rana japonica and R. brevipoda, raised from UV irradiated eggs.

    PubMed

    Shirane, T

    1982-10-10

    The gonadal differentiation of anurans, Rana japonica and R. brevipoda, was examined in animals raised from eggs which had been irradiated at the vegetal hemisphere with UV (9300 erg/mm2) at the 2-cell stage. In R. japonica about 70% of the larvae at stage I from the pressed and UV-irradiated eggs were germ cell free, but at a stage immediately after metamorphosis all animals had at least some germ cells, although their gonads often were extremely small and poorly differentiated. When male animals matured sexually, many of them had abnormal gonads. However, all of them were shown by artificial means to be capable of fertilization. In the nonpressed and irradiated group, no larvae were germ cell free and the animals immediately after metamorphosis showed nearly normal gonadal differentiation except for the presence of a few degenerate oocytes in the ovaries. The results in R. brevipoda were basically similar to those in R. japonica. In both species, sex ratios were determined at two stages, the first immediately after metamorphosis and the other when the animals matured, as based on gonad morphology and histology and on external sexually dimorphic characters as well. Sex ratios at these two stages in frogs from the pressed and irradiated eggs differed markedly in R. brevipoda. The ratio was normal at metamorphosis but high M/F ratios occurred when animals became mature. That sex reversal took place in this species as well as in R. japonica (in which sex-ratio deviation was not statistically significant) was supported by the sex ratios of the progenies of these supernumerary males.

  2. Plasticity of auditory medullary-midbrain connectivity across metamorphic development in the bullfrog, Rana catesbeiana.

    PubMed

    Horowitz, Seth S; Chapman, Judith A; Simmons, Andrea Megela

    2007-01-01

    On the basis of patterns of anterograde, retrograde, and bi-directional transport of tracers from both the superior olivary nucleus (SON) and the torus semicircularis (TS), we report anatomical changes in brainstem connectivity across metamorphic development in the bullfrog, Rana catesbeiana. In early and late stages of larval development (Gosner stages 25-37), anterograde or bi-directional tracers injected into the SON produce terminal/fiber label in the contralateral SON and in the ipsilateral TS. Between stages 38-41 (deaf period), only sparse or no terminal/fiber label is visible in these target nuclei. During metamorphic climax (stages 42-46), terminal/fiber label reappears in both the contralateral SON and in the ipsilateral TS, and now also in the contralateral TS. Injections of retrograde tracers into the SON fail to label cell bodies in the ipsilateral TS in deaf period animals, mirroring the previously-reported failure of retrograde transport from the TS to the ipsilateral SON during this developmental time. Bilateral cell body label emerges in the dorsal medullary nucleus and the lateral vestibular nucleus bilaterally as a result of SON transport during the late larval period, while cell body label in the contralateral TS emerges during climax. At all larval stages, injections into the SON produce anterograde and retrograde label in the medial vestibular nucleus bilaterally. These data show anatomical stability in some pathways and plasticity in others during larval development, with the most dramatic changes occurring during the deaf period and metamorphic climax. Animals in metamorphic climax show patterns of connectivity similar to that of froglets and adults, indicating the maturation during climax of central anatomical substrates for hearing in air.

  3. Shading as a Control Method for Invasive European Frogbit (Hydrocharis morsus-ranae L.)

    PubMed Central

    Zhu, Bin; Ellis, Michael S.; Fancher, Kelly L.; Rudstam, Lars G.

    2014-01-01

    Invasive European frogbit (Hydrocharis morsus-ranae L.) has negative environmental and economic impacts in North American water bodies. It is therefore important to develop effective management tools to control this invasive species. This study investigated shading as a control method for European frogbit in both greenhouse and lake mesocosm experiments. A series of shade treatments (0%, 50%, 60%, 70%, 80%, and 100%) were tested in the greenhouse for three weeks. Results showed that the 100% shade was most effective at controlling European frogbit, and other shade treatments greater than 50% were less effective, reducing frogbit biomass up to 38.2%. There were no differences found in temperature between treatments, but dissolved oxygen decreased as shading increased. A lake mesocosm experiment utilizing 0% shade, 70% shade, and 100% shade treatments was performed in a sheltered inlet of Oneida Lake in New York State for over one month. Resulting European frogbit biomass was significantly (25 times) less in areas treated with the 70% shade and nearly zero with the 100% shade. Shading did not affect temperature but improved DO conditions. Results on the shading effects on submerged macrophytes were not conclusive: no significant differences in changes in species richness and abundance between the three groups at the end of studied period suggested no shading effects; significant differences between the beginning and end communities in the 70% shade and the 100% shade but not in the control group indicated significant impacts of shading. This study is the first one to investigate shading as a control method for European frogbit and it is concluded that a moderately high density shade can effective remove European frogbit likely with minor impacts on the environment. More experiments with larger scales and longer time periods are recommended for further investigation. PMID:24886916

  4. Cardiovascular responses to catecholamines at 12 degrees C in the American bullfrog (Rana catesbeiana).

    PubMed

    Herman, C A; Robleto, D O; Mata, P L; Heller, R S

    1986-10-01

    The effects of epinephrine, norepinephrine, phenylephrine, and isoproterenol on blood pressure and heart rate were studied in cannulated American bullfrogs, Rana catesbeiana. The bullfrogs were chronically cannulated with a T cannula in the right sciatic artery. In warm-acclimated (22 degrees C) bullfrogs, preinjection mean systemic arterial pressure (SAP) prior to experimental treatment was 13.1 +/- 0.7 mm Hg. Preinjection heart rate was 34.8 +/- 1.8 beats per minute. These parameters were lower in cold-acclimated (12 degrees C) bullfrogs. Cold-acclimated animals had mean SAP values of 8.2 +/- 0.3 mm Hg, and heart rate was 11.1 +/- 1.1 beats per minute. Epinephrine, norepinephrine, and phenylephrine increased blood pressure to an equivalent degree in warm- and cold-acclimated animals. Dose-related decreases in heart rate in response to these catecholamines were observed in warm- but not in cold-acclimated bullfrogs. Warm-acclimated animals were more responsive to isoproterenol from 0.03 micrograms/kg body weight (bw) to 10 micrograms/kg bw than were cold-acclimated animals. The response to isoproterenol was effectively blocked by propranolol (5 mg/kg bw) in both warm- and cold-acclimated animals. Propranolol alone decreased mean SAP in both warm- and cold-acclimated animals, suggesting blockade of endogenous sympathetic activity. Beta receptor response thus appears diminished, but not absent at 12 degrees C. However, the alpha receptors responsible for elevation of blood pressure equally responsive at 12 degrees and 22 degrees C.

  5. Clinical signs, pathology and dose-dependent survival of adult wood frogs, Rana sylvatica, inoculated orally with frog virus 3 Ranavirus sp., Iridoviridae.

    PubMed

    Forzn, Mara J; Jones, Kathleen M; Vanderstichel, Raphal V; Wood, John; Kibenge, Frederick S B; Kuiken, Thijs; Wirth, Wytamma; Ariel, Ellen; Daoust, Pierre-Yves

    2015-05-01

    Amphibian populations suffer massive mortalities from infection with frog virus 3 FV3, genus Ranavirus, family Iridoviridae, a pathogen also involved in mortalities of fish and reptiles. Experimental oral infection with FV3 in captive-raised adult wood frogs, Rana sylvatica Lithobates sylvaticus, was performed as the first step in establishing a native North American animal model of ranaviral disease to study pathogenesis and host response. Oral dosing was successful LD50 was 10(2.93 2.423.44) p.f.u. for frogs averaging 35mm in length. Onset of clinical signs occurred 614days post-infection p.i. median 11 days p.i. and time to death was 1014 days p.i. median 12 days p.i.. Each tenfold increase in virus dose increased the odds of dying by 23-fold and accelerated onset of clinical signs and death by approximately 15. Ranavirus DNA was demonstrated in skin and liver of all frogs that died or were euthanized because of severe clinical signs. Shedding of virus occurred in faeces 710 days p.i. 34.5days before death and skin sheds 10 days p.i. 01.5days before death of some frogs dead from infection. Most common lesions were dermal erosion and haemorrhages haematopoietic necrosis in bone marrow, kidney, spleen and liver and necrosis in renal glomeruli, tongue, gastrointestinal tract and urinary bladder mucosa. Presence of ranavirus in lesions was confirmed by immunohistochemistry. Intracytoplasmic inclusion bodies probably viral were present in the bone marrow and the epithelia of the oral cavity, gastrointestinal tract, renal tubules and urinary bladder. Our work describes a ranaviruswood frog model and provides estimates that can be incorporated into ranavirus disease ecology models.

  6. Mass mortality associated with a frog virus 3-like Ranavirus infection in farmed tadpoles Rana catesbeiana from Brazil

    PubMed Central

    Mazzoni, Rolando; de Mesquita, Albenones José; Fleury, Luiz Fernando F.; de Brito, Wilia Marta Elsner Diederichsen; Nunes, Iolanda A.; Robert, Jacques; Morales, Heidi; Coelho, Alexandre Siqueira Guedes; Barthasson, Denise Leão; Galli, Leonardo; Catroxo, Marcia H. B.

    2010-01-01

    Ranviruses (Iridoviridae) are increasingly associated with mortality events in amphibians, fish, and reptiles. They have been recently associated with mass mortality events in Brazilian farmed tadpoles of the American bullfrog Rana catesbeiana Shaw. 1802. The objectives of the present study were to further characterize the virus isolated from sick R. catesbeiana tadpoles and confirm the etiology in these outbreaks. Sick tadpoles were collected in 3 farms located in Goiás State, Brazil, from 2003 to 2005 and processed for virus isolation and characterization, microbiology, histopathology, and parasitology. The phylogenetic relationships of Rana catesbeiana ranavirus (RCV-BR) with other genus members was investigated by PCR with primers specific for the major capsid protein gene (MCP) and the RNA polymerase DNA-dependent gene (Pol II). Sequence analysis and multiple alignments for MCP products showed >99% amino acid identity with other ranaviruses, while Pol II products showed 100% identity. Further diagnostics of the pathology including histology and transmission electron microscopy confirmed the viral etiology of these mass deaths. As for as we know, this is the first report of a ranaviral infection affecting aquatic organisms in Brazil. Additionally, our results suggest that American bullfrogs may have served as a vector of transmission of this virus, which highlights the potential threat of amphibian translocation in the world distribution of pathogens. PMID:20066953

  7. Response to pinealectomy and blinding in vitellogenic female frogs (Rana perezi) subjected to high temperature in autumn.

    PubMed

    Alonso-Gómez, A L; Tejera, M; Alonso-Bedate, M; Delgado, M J

    1990-01-01

    The present experiments were carried out to investigate the effects of pinealectomy and bilateral enucleation on the ovarian activity in Rana perezi frogs maintained in 12-h light--12-h dark photoperiod and 20 +/- 1 degrees C during the vitellogenetic growth in late autumn. These environmental conditions, mainly temperature, induce a gonadal and metabolic response similar to that observed in the natural habitat in summer: a marked ovarian follicular regression, a depletion of the energetic resources from fat bodies and liver, and a minimum in oestradiol circulating levels. This response is partially blocked by pinealectomy and blinding. Protein phosphorus, as an index of vitellogenic proteins, and total ovary lipid content were significantly higher in pinealectomized and blinded frogs with respect to sham-operated animals. Likewise, oestradiol concentrations showed a significant increase during the dark phase of the daily photocycle in pinealectomized and blinded animals. From our results, we can suggest that the arrest of vitellogenesis, the depletion of energetic resources, and the regulation of oestradiol levels induced by the high temperature in Rana perezi frogs can be influenced, at least in part, by the pineal complex and lateral eyes.

  8. Structural analysis of oligosaccharide-alditols released by reductive beta-elimination from oviducal mucins of Rana dalmatina.

    PubMed

    Morelle, W; Guyétant, R; Strecker, G

    1998-01-01

    The O-linked oligosaccharides of the jelly coat surrounding the eggs of Rana dalmantina were released by alkaline borohydride treatment. Low-molecular-mass, monosialyl oligosaccharide-alditols were isolated by anion-exchange chromatography and fractionated by consecutive normal-phase high-performance liquid chromatography on a silica-based alkylamine column. The structures of the oligosaccharide-alditols were determined by 400-MHz 1H-NMR spectroscopy in combination with matrix assisted laser desorption ionization-time of flight analysis. The five structures were identified range in size from trisaccharides to hexasaccharides, possessing a core consisting of Gal(beta 1-3)GalNAc-ol (core type 1). Novel oligosaccharide-alditols are: [formula: see text] The carbohydrate chains isolated from Rana dalmatina are different from those found in other amphibian species, in which the presence of species-specific material has been characterized. Since the role of carbohydrates appears more and more apparent during the fertilization process, the biodiversity of the O-linked oligosaccharides could support such a biological role.

  9. Speciation in the Rana chensinensis species complex and its relationship to the uplift of the Qinghai-Tibetan Plateau.

    PubMed

    Zhou, Wei-Wei; Wen, Yang; Fu, Jinzhong; Xu, Yong-Biao; Jin, Jie-Qiong; Ding, Li; Min, Mi-Sook; Che, Jing; Zhang, Ya-Ping

    2012-02-01

    Speciation remains a fundamental issue in biology. Herein, we report an investigation into speciation in the Rana chensinensis species complex using DNA sequence data from one mitochondrial and five nuclear genes. A phylogenetic analysis of the data revealed four major clades in the complex, and each of them was found to likely represent a species, including one cryptic species. Ecological niche models were generated from 19 climatic variables for three of the four major clades, which were represented by widespread sampling, including R. chensinensis, Rana kukunoris and the potential cryptic species. Each clade is associated with a unique ecological unit, and this indicates that ecological divergence probably drove speciation. Ecological divergence is likely related to the late Cenozoic orogenesis of the Qinghai-Tibetan Plateau. In addition, gene flow between species was detected but only in peripheral portions of the ranges of the four major clades, thus likely had little influence on the speciation processes. Discordances between mitochondrial and nuclear genes were also found; the nominal species, R. chensinensis, contains multiple maternal clades, suggesting potential mitochondrial introgression between R. chensinensis and R. kukunoris.

  10. Expression of P450arom and Estrogen Receptor Alpha in the Oviduct of Chinese Brown Frog (Rana dybowskii) during Prehibernation

    PubMed Central

    Weng, Ji; Liu, Yuning; Xu, Ying; Hu, Ruiqi; Zhang, Haolin; Sheng, Xia; Watanabe, Gen; Taya, Kazuyoshi; Weng, Qiang; Xu, Meiyu

    2015-01-01

    One specific physiological phenomenon of Chinese brown frog (Rana dybowskii) is that its oviduct expands prior to hibernation instead of expanding during the breeding period. In this study, we investigated the expression of P450arom and estrogen receptors α and β (ERα and ERβ) in the oviduct of Rana dybowskii during the breeding period and prehibernation. The results of the present study showed that there were significant differences in both oviductal weight and size with values markedly higher in prehibernation than in the breeding period. P450arom was observed in stromal tissue in both the breeding period and prehibernation. ERα was expressed in stromal tissue and epithelial cells in both periods, whereas ERβ could not be detected. The mean protein and mRNA levels of P450arom and ERα were significantly higher in prehibernation as compared to the breeding period. Besides, oviductal content of 17β-estradiol was also higher in prehibernation than in the breeding period. These results suggested that estrogen may play autocrine/paracrine roles mediated by ERα in regulating the oviductal hypertrophy during prehibernation. PMID:25802518

  11. Essential oils of indigenous in Greece six Juniperus taxa: chemical composition and larvicidal activity against the West Nile virus vector Culex pipiens.

    PubMed

    Vourlioti-Arapi, F; Michaelakis, A; Evergetis, E; Koliopoulos, G; Haroutounian, S A

    2012-05-01

    The chemical composition of 14 essential oils (EOs), obtained from various parts (leaves, fruits, wood) of the six indigenous in Greece Juniperus family taxa, was determined by GC and GC/MS analysis. The insecticidal properties of these EOs were evaluated against Culex pipiens L. larvae of 3rd and early 4th instars, in order to delineate the relationship between the phytochemical content of the EOs and their larvicidal activities. The analytical data indicated that the EOs mainly consisted of monoterpenes, mostly cyclic and only occasionally aliphatic, and to a lesser percent, of diterpenes. The larvicidal bioassays against C. pipiens larvae revealed that the most active EO was derived from the wood of Juniperus drupacea and contains mainly non-oxygenated monoterpenes and a significant amount of diterpenes, displaying the highest chemodiversity. Its initial LC(50) value was 26.47 mg L(-1). On the contrary, the EO isolated from J. phoenicea berries, which consisted of monoterpenes (non-oxygenated, cyclic), was the less active displaying an LC(50) value of 96.69 mg L(-1). In respect to the contained phytochemicals, myrcene was assayed as the most toxic, displaying an LC(50) value of 33.83 mg L(-1), while the four isomers of pinene abundant in all EOs were less active exhibiting LC(50) values ranging from 70.40 to 94.88 mg L(-1). Results herein reveal that the EOs isolated from the studied Juniperus family taxa represent an inexpensive source of natural mosquito control mixtures.

  12. Modeling the impact of variable climatic factors on the crossover of Culex restauns and Culex pipiens (Diptera: culicidae), vectors of West Nile virus in Illinois.

    PubMed

    Kunkel, Kenneth E; Novak, Robert J; Lampman, Richard L; Gu, Weidong

    2006-01-01

    The aim of this study was to model the impact of temperature on the timing of the seasonal shift in relative proportion of Culex restuans Theobald and Culex pipiens L. in Illinois. The temporal pattern of West Nile virus (WNV) and St. Louis encephalitis virus transmission in the midwest exhibits a late summer to early fall peak in activity, which parallels the temporal increase in the abundance of Cx. pipiens. The daily number of egg rafts oviposited by each species has been monitored at multiple surveillance sites in Urbana-Champaign in central Illinois for more than 13 years. The time when the two Culex species are in equal abundance (crossover) varies considerably from year to year. Our investigation of several thermal measures indicated that this variation was related in large part to climatic conditions with warmer (cooler) temperatures correlated to earlier (later) crossover dates. Models based on degree days and the number of days in which the daily maximum temperature exceeded an upper temperature threshold explained more than 60% of the variance in crossover dates. In contrast, models based on the number of days in which the daily minimum temperature exceeded a lower temperature threshold explained no more than 52% of the variance. An evaluation of these models demonstrated that they provide relatively simple and accurate estimates of crossover date from daily temperature data, a necessary component for developing an overall climatic index for the risk of WNV transmission in Illinois.

  13. Attraction of Culex pipiens/restuans (Diptera: Culicidae) mosquitoes to bird uropygial gland odors at two elevations in the Niagara region of Ontario.

    PubMed

    Russell, Curtis B; Hunter, Fiona F

    2005-05-01

    In an effort to determine whether female Culex pipiens L. and Culex restuans Theobald mosquitoes (Diptera: Culicidae) are attracted to crow, Corvus brachyrhynchus, uropygial gland secretions, CDC miniature light traps (baited with CO2 but with the lights removed) were placed at approximately 1.5- and 5-m elevations, in 10 trees in awoodlot near Niagara Falls, Canada. These traps were assigned either a bird odor or a blank control. Bird odors were created by attaching cotton swabs coated with crow uropygial gland secretions to the trap intake. A significantly greater number of Cx. pipiens/ restuans were found in the 5-m traps compared with 1.5-m traps, with a significant number attracted to the bird odor over the no odor traps at the 5-m elevation, but not at 1.5 m. We also found more Aedes vexans (Meigen) in the 1.5-m traps than the 5-m traps; however, presence or absence of bird odor did not influence the distribution of Ae. vexans.

  14. Regulation of 5'-adenosine monophosphate deaminase in the freeze tolerant wood frog, Rana sylvatica

    PubMed Central

    Dieni, Christopher A; Storey, Kenneth B

    2008-01-01

    Background The wood frog, Rana sylvatica, is one of a few vertebrate species that have developed natural freeze tolerance, surviving days or weeks with 65–70% of its total body water frozen in extracellular ice masses. Frozen frogs exhibit no vital signs and their organs must endure multiple stresses, particularly long term anoxia and ischemia. Maintenance of cellular energy supply is critical to viability in the frozen state and in skeletal muscle, AMP deaminase (AMPD) plays a key role in stabilizing cellular energetics. The present study investigated AMPD control in wood frog muscle. Results Wood frog AMPD was subject to multiple regulatory controls: binding to subcellular structures, protein phosphorylation, and effects of allosteric effectors, cryoprotectants and temperature. The percentage of bound AMPD activity increased from 20 to 35% with the transition to the frozen state. Bound AMPD showed altered kinetic parameters compared with the free enzyme (S0.5 AMP was reduced, Hill coefficient fell to ~1.0) and the transition to the frozen state led to a 3-fold increase in S0.5 AMP of the bound enzyme. AMPD was a target of protein phosphorylation. Bound AMPD from control frogs proved to be a low phosphate form with a low S0.5 AMP and was phosphorylated in incubations that stimulated PKA, PKC, CaMK, or AMPK. Bound AMPD from frozen frogs was a high phosphate form with a high S0.5 AMP that was reduced under incubation conditions that stimulated protein phosphatases. Frog muscle AMPD was activated by Mg·ATP and Mg·ADP and inhibited by Mg·GTP, KCl, NaCl and NH4Cl. The enzyme product, IMP, uniquely inhibited only the bound (phosphorylated) enzyme from muscle of frozen frogs. Activators and inhibitors differentially affected the free versus bound enzyme. S0.5 AMP of bound AMPD was also differentially affected by high versus low assay temperature (25 vs 5°C) and by the presence/absence of the natural cryoprotectant (250 mM glucose) that accumulates during freezing

  15. Evaluation of the effects of titanium dioxide nanoparticles on cultured Rana catesbeiana tailfin tissue.

    PubMed

    Hammond, S Austin; Carew, Amanda C; Helbing, Caren C

    2013-01-01

    Nanoparticles (NPs), materials that have one dimension less than 100 nm, are used in manufacturing, health, and food products, and consumer products including cosmetics, clothing, and household appliances. Their utility to industry is derived from their high surface-area-to-volume ratios and physico-chemical properties distinct from their bulk counterparts, but the near-certainty that NPs will be released into the environment raises the possibility that they could present health risks to humans and wildlife. The thyroid hormones (THs), thyroxine, and 3,3',5-triiodothyronine (T3), are involved in development and metabolism in vertebrates including humans and frogs. Many of the processes of anuran metamorphosis are analogous to human post-embryonic development and disruption of TH action can have drastic effects. These shared features make the metamorphosis of anurans an excellent model for screening for endocrine disrupting chemicals (EDCs). We used the cultured tailfin (C-fin) assay to examine the exposure effects of 0.1-10 nM (~8-800 ng/L) of three types of ~20 nm TiO2 NPs (P25, M212, M262) and micron-sized TiO2 (μ TiO2) ±10 nM T3. The actual Ti levels were 40.9-64.7% of the nominal value. Real-time quantitative polymerase chain reaction (QPCR) was used to measure the relative amounts of mRNA transcripts encoding TH-responsive THs receptors (thra and thrb) and Rana larval keratin type I (rlk1), as well as the cellular stress-responsive heat shock protein 30 kDa (hsp30), superoxide dismutase (sod), and catalase (cat). The levels of the TH-responsive transcripts were largely unaffected by any form of TiO2. Some significant effects on stress-related transcripts were observed upon exposure to micron-sized TiO2, P25, and M212 while no effect was observed with M262 exposure. Therefore, the risk of adversely affecting amphibian tissue by disrupting TH-signaling or inducing cellular stress is low for these compounds relative to other previously-tested NPs.

  16. Sound and vibration sensitivity of VIIIth nerve fibers in the grassfrog, Rana temporaria.

    PubMed

    Christensen-Dalsgaard, J; Jørgensen, M B

    1996-10-01

    We have studied the sound and vibration sensitivity of 164 amphibian papilla fibers in the VIIIth nerve of the grassfrog, Rana temporaria. The VIIIth nerve was exposed using a dorsal approach. The frogs were placed in a natural sitting posture and stimulated by free-field sound. Furthermore, the animals were stimulated with dorso-ventral vibrations, and the sound-induced vertical vibrations in the setup could be canceled by emitting vibrations in antiphase from the vibration exciter. All low-frequency fibers responded to both sound and vibration with sound thresholds from 23 dB SPL and vibration thresholds from 0.02 cm/s2. The sound and vibration sensitivity was compared for each fiber using the offset between the rate-level curves for sound and vibration stimulation as a measure of relative vibration sensitivity. When measured in this way relative vibration sensitivity decreases with frequency from 42 dB at 100 Hz to 25 dB at 400 Hz. Since sound thresholds decrease from 72 dB SPL at 100 Hz to 50 dB SPL at 400 Hz the decrease in relative vibration sensitivity reflects an increase in sound sensitivity with frequency, probably due to enhanced tympanic sensitivity at higher frequencies. In contrast, absolute vibration sensitivity is constant in most of the frequency range studied. Only small effects result from the cancellation of sound-induced vibrations. The reason for this probably is that the maximal induced vibrations in the present setup are 6-10 dB below the fibers' vibration threshold at the threshold for sound. However, these results are only valid for the present physical configuration of the setup and the high vibration-sensitivities of the fibers warrant caution whenever the auditory fibers are stimulated with free-field sound. Thus, the experiments suggest that the low-frequency sound sensitivity is not caused by sound-induced vertical vibrations. Instead, the low-frequency sound sensitivity is either tympanic or mediated through bone conduction or sound

  17. Evaluation of the effects of titanium dioxide nanoparticles on cultured Rana catesbeiana tailfin tissue

    PubMed Central

    Hammond, S. Austin; Carew, Amanda C.; Helbing, Caren C.

    2013-01-01

    Nanoparticles (NPs), materials that have one dimension less than 100 nm, are used in manufacturing, health, and food products, and consumer products including cosmetics, clothing, and household appliances. Their utility to industry is derived from their high surface-area-to-volume ratios and physico-chemical properties distinct from their bulk counterparts, but the near-certainty that NPs will be released into the environment raises the possibility that they could present health risks to humans and wildlife. The thyroid hormones (THs), thyroxine, and 3,3′,5-triiodothyronine (T3), are involved in development and metabolism in vertebrates including humans and frogs. Many of the processes of anuran metamorphosis are analogous to human post-embryonic development and disruption of TH action can have drastic effects. These shared features make the metamorphosis of anurans an excellent model for screening for endocrine disrupting chemicals (EDCs). We used the cultured tailfin (C-fin) assay to examine the exposure effects of 0.1–10 nM (~8–800 ng/L) of three types of ~20 nm TiO2 NPs (P25, M212, M262) and micron-sized TiO2 (μ TiO2) ±10 nM T3. The actual Ti levels were 40.9–64.7% of the nominal value. Real-time quantitative polymerase chain reaction (QPCR) was used to measure the relative amounts of mRNA transcripts encoding TH-responsive THs receptors (thra and thrb) and Rana larval keratin type I (rlk1), as well as the cellular stress-responsive heat shock protein 30 kDa (hsp30), superoxide dismutase (sod), and catalase (cat). The levels of the TH-responsive transcripts were largely unaffected by any form of TiO2. Some significant effects on stress-related transcripts were observed upon exposure to micron-sized TiO2, P25, and M212 while no effect was observed with M262 exposure. Therefore, the risk of adversely affecting amphibian tissue by disrupting TH-signaling or inducing cellular stress is low for these compounds relative to other previously-tested NPs. PMID

  18. [Morphological and physiological characterization of fiber types in the iliofibular muscle of Rana esculenta].

    PubMed

    Dauber, W

    1977-01-01

    In both longitudinal and cross sections of the M. iliofibularis of Rana esculenta three types of muscle fibres are identified by means of light and electron microscopy. These fibretypes called A-, B- and C-fibres are according to the fibres of m. rectus abdominis of the frog. They can be compared with the fibres of the m. rectus abdominis of rat and mouse. But there is another distribution of the fibretypes A, B and C in the m. iliofibularis and in the m. rectus abdominis. The m. iliofibularis is divided into two parts called "Tonusbündel" and "nichttonischer Teil" by means of their reaction to acetylcholine. The surface of the "Tonusbündel" consists of A-, B- and C-fibres while its inside is onlyformed by A- and B-fibres. They continue the "Tonusbündel" in the "nichttonischer Teil". This part chiefly consists of A-fibres. In cross sections their myofibrils are larger in their extent than the A-fibres known before. Therefore the A-fibretype has to be distinguished into two A-fibres: A1 and A2. The new one is called A2-fibre. A1-fibre is described in the "Tonusbündel" and in further investigations. The difference between the two fibres can be understood as a greater manifestation of power of the A1-fibre. The surface of the "nichttonischer Teil" of the m. iliofibularis consists of A2-fibres which easily could be found opposite the "Tonusbündel". At this point in contrary to the "Tonusbündel" could be found a defined morphological substrate for physiological investigations. The different reactions of "Tonusbündel" and "nichttonischer Teil" to acetylcholine could only be explained by the sum of reactions of all fibretypes in each bundle in correspondence with the reaction of the fibres in the neighbour bundle. But their different behaviour by summer- and winterfrogs is unknown. Therefore it is to discuss whether it is allowed to refer generally the results to "muscle" or "musclefibre" got from frogs living in cooled rooms. It is known in literature that not all

  19. Endocrine effects of environmental pollution on Xenopus laevis and Rana temporaria.

    PubMed

    Bögi, C; Schwaiger, J; Ferling, H; Mallow, U; Steineck, C; Sinowatz, F; Kalbfus, W; Negele, R D; Lutz, I; Kloas, W

    2003-10-01

    To determine the capacity of sewage treatment work effluents to disrupt the endocrine system under semifield conditions, two amphibian species, Xenopus laevis and Rana temporaria, were exposed to the effluent of a regional sewage treatment plant in South Bavaria during larval development until completion of metamorphosis. Exposure was carried out in river water (Würm) as a reference, and a 1:12-mixture sewage effluent representing the real situation on the spot, and in a higher concentration of sewage using a 1:2 mixture. An accidental impact of industrial wastewater into the reference and dilution medium, Würm, which was caused by a spate in the respective area during the sensitive period of sex differentiation of amphibian larvae, is assumed to be responsible for the relatively high percentage of females observed by histological analysis in all treatment groups. All of these values were higher than those determined in controls exposed to artificial tap water in laboratory experiments conducted in a comparable study design. Sex ratios between species, revealed by the semifield study with decreasing portions of females from control to 1:12 to 1:2, were strongly correlated. Determination of biomarker-mRNA-levels in Xenopus liver using semiquantitative RT-PCR at the end of the experimental phase, when exposure regime has turned into the initially expected situation with the highest load of potential estrogens in the effluent, followed by 1:2 and 1:12 mixture, resulted in a significant increase of Vitellogenin-mRNA in female juveniles exposed to the highest portion of sewage, whereas expression of both androgen and estrogen receptor-mRNA showed no clear differences. The results concerning the induction of estrogenic biomarkers are in accordance with our findings for estrogen receptor binding of sample extracts from the Würm and sewage taken in parallel at the end of the experiment, when sewage extracts possessed a much higher ability to displace [3H]estradiol from

  20. Identification and characterisation of a novel antimicrobial polypeptide from the skin secretion of a Chinese frog (Rana chensinensis).

    PubMed

    Jin, Li L; Song, Shu S; Li, Qiang; Chen, Yu H; Wang, Qiu Y; Hou, Sheng T

    2009-06-01

    Amphibians secrete small antimicrobial polypeptides from their skin that have been explored as alternatives to conventional antibiotics. In this study, mass spectrometry was used to identify and characterise protein secretions from the skin of a Chinese frog, Rana chensinensis. The skin of this kind of frog has been used in traditional Chinese medicine for centuries as a remedy against inflammation. A novel antimicrobial peptide was identified and the characteristics of this peptide were analysed using far-ultraviolet circular dichroism. When dissolved in aqueous solution, the peptide displayed a high level of random coil structure, in contrast to a more ordered alpha-helical structure when dissolved in 50% trifluoroethanol. Functional studies showed that this peptide has potent antimicrobial activity both against Gram-positive and Gram-negative bacteria and has extremely low haemolytic activity to human red blood cells. Taken together, these studies suggest that this novel peptide can be further developed as an antimicrobial agent.

  1. The function of fat bodies in relation to the hypothalamo-hypophyseal-gonadal axis in the frog, Rana esculenta.

    PubMed

    Chieffi, G; Rastogi, R K; Iela, L; Milone, M

    1975-08-18

    In this study the authors have tried to furnish experimental support for the importance of fat bodies in the normal functioning of the hypothalamo-hypophyseal-gonadal system of the male frog, Rana esculenta. These experiments have shown a hypothalamo-hypophyseal control of the mobilization of fat body contents, directly involved in the control of testicular activity. Furthermore it is proposed that the fat body contents are released into the testis through direct vascular contacts between the two organs. We suggest that the A1 cells (lactotrophs) and/or B2 cells (FSH-gonadotrops) of the pars distalis gonadotropins are incapable of stimulating the testis in the absence of fat bodies. In the light of these results a scheme has been put forward showing the position of fat bodies in the hypothalamo-hypophyseal-gonadal axis of the frog.

  2. Influence of sex and breeding condition on microhabitat selection and diet in the pig frog Rana grylio

    SciTech Connect

    Lamb, T.

    1984-04-01

    A 14-month study was conducted on the pig frog (Rana grylio) in SW Georgia. This species has a prolonged breeding season as males call from late March to September. Mature spermatozoa were present in the testes year-round, though seasonal testicular changes were detectable with spermatogenesis reaching a peak in June. Females contained mature ova from April through July and development of the following year's ova began in August. Stomachs of 122 postlarval specimens contained mainly anthropods. Coleoptera, Decopoda (Procambarus) and Odonata accounted for the majority of individual prey items, constituting 24.3, l9.8 and 11.9%, respectively. Intersexual dietary differences were apparent among adult frogs during the breeding season; variation in diet was strongly influenced by behavioral and habitat differences at this time.

  3. Short-term occupancy and abundance dynamics of the Oregon spotted frog (Rana pretiosa) across its core range

    USGS Publications Warehouse

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

    2014-01-01

    The Oregon spotted frog (Rana pretiosa) occupies only a fraction of its original range and is listed as Threatened under the Endangered Species Act. We surveyed 93 sites in a rotating frame design (2010–13) in the Klamath and Deschutes Basins, Oregon, which encompass most of the species’ core extant range. Oregon spotted frogs are declining in abundance and probability of site occupancy. We did not find an association between the probability that Oregon spotted frogs disappear from a site (local extinction) and any of the variables hypothesized to affect Oregon spotted frog occupancy. This 4-year study provides baseline data, but the 4-year period was too short to draw firm conclusions. Further study is essential to understand how habitat changes and management practices relate to the status and trends of this species.

  4. Endogenous peroxidase activity in brush cell-like cells in the large intestine of the bullfrog tadpole, Rana catesbeiana.

    PubMed

    Sugimoto, K; Ichikawa, Y; Nakamura, I

    1983-01-01

    A special cell type was identified in the mucosal epithelium of the large intestine of the tadpole of the bullfrog, Rana catesbeiana. It is a slender, columnar cell, with a dark, basally situated nucleus. By electron microscopy the cell displays prominent bundles of filaments emerging from each microvillus and extending deep into the cytoplasm without ending in the terminal web. It has longer and more crowded microvilli than the absorptive cell. The specialized cell is also characterized by the presence of many apical vesicles and numerous subapical dense bodies. These cytological features suggest that it may be a brush cell (Rhodin and Dalhamn 1956). These cells displayed endogenous peroxidase activity in smooth and rough endoplasmic reticulum, in the well-developed Golgi apparatus and in apical vesicles. Furthermore, peroxidase reaction product was frequently observed on their luminal surface membrane. These findings suggest that the brush cell in the large intestine of the bullfrog tadpole may be a secretory cell.

  5. Effects of bilateral and unilateral ophthalmectomy on plasma melatonin in Rana tadpoles and froglets under various experimental conditions.

    PubMed

    Wright, Mary L; Francisco, Lucy L; Scott, Jessica L; Richardson, Shaun E; Carr, James A; King, Amy B; Noyes, Arielle G; Visconti, Rachael F

    2006-06-01

    The effect of ophthalmectomy (enucleation) on plasma melatonin in Rana tadpoles and froglets was studied under various experimental conditions to determine if ocular melatonin is released into the circulation from the eyes and to study the factors which might affect this process. Where operations occurred in early or mid-photophase on a 12 light:12 dark (12L:12D) cycle (light onset at 08:00 h), sampling in mid-light and mid-dark revealed that scotophase plasma melatonin was reduced at all developmental stages, with the more significant effects occurring before metamorphic climax. Experiments sampling prometamorphic tadpoles six times in a 24h period on 18L:6D, 12L:12D, or 6L:18D five days after enucleation also showed a significant lowering of plasma melatonin in the dark, so that the scotophase peak was virtually eliminated on all the LD cycles. These findings indicated that the reduction in plasma melatonin after bilateral eye removal was independent of the LD cycle and the metamorphic stage, and that it abolished the diel melatonin rhythm at the expense of the scotophase peak. Experiments carried out for 5 weeks suggested that compensatory secretion of melatonin by other organs after eye removal might partially restore the plasma melatonin level over time. Unilateral ophthalmectomy tended to reduce, but not eliminate, the night peak of plasma melatonin, and did not result in a compensatory increase in ocular melatonin in the remaining eye. Ophthalmectomized tadpoles exhibited darkening of the skin after the operation, which was not associated with a significant change in pituitary alpha-melanotropin. The findings overall indicate that the eyes in Rana tadpoles and froglets contribute up to somewhat over one-half of the circulating melatonin, particularly during the scotophase, and provide experimental evidence for ocular secretion into the blood for the first time in the Amphibia.

  6. A new species of leopard frog (Anura: Ranidae) from the urban northeastern US

    PubMed Central

    Newman, Catherine E.; Feinberg, Jeremy A.; Rissler, Leslie J.; Burger, Joanna; Shaffer, H. Bradley

    2014-01-01

    Past confusion about leopard frog (genus Rana) species composition in the Tri-State area of the US that includes New York (NY), New Jersey (NJ), and Connecticut (CT) has hindered conservation and management efforts, especially where populations are declining or imperiled. We use nuclear and mitochondrial genetic data to clarify the identification and distribution of leopard frog species in this region. We focus on four problematic frog populations of uncertain species affiliation in northern NJ, southeastern mainland NY, and Staten Island to test the following hypotheses: (1) they are conspecific with Rana sphenocephala or R. pipiens, (2) they are hybrids between R. sphenocephala and R. pipiens, or (3) they represent one or more previously undescribed cryptic taxa. Bayesian phylogenetic and cluster analyses revealed that the four unknown populations collectively form a novel genetic lineage, which represents a previously undescribed cryptic leopard frog species, Rana sp. nov. Statistical support for R. sp. nov. was strong in both the Bayesian (pp = 1.0) and maximum-likelihood (bootstrap = 99) phylogenetic analyses as well as the Structure cluster analyses. While our data support recognition of R. sp. nov. as a novel species, we recommend further study including fine-scaled sampling and ecological, behavioral, call, and morphological analyses before it is formally described. PMID:22321689

  7. A new species of leopard frog (Anura: Ranidae) from the urban northeastern US.

    PubMed

    Newman, Catherine E; Feinberg, Jeremy A; Rissler, Leslie J; Burger, Joanna; Shaffer, H Bradley

    2012-05-01

    Past confusion about leopard frog (genus Rana) species composition in the Tri-State area of the US that includes New York (NY), New Jersey (NJ), and Connecticut (CT) has hindered conservation and management efforts, especially where populations are declining or imperiled. We use nuclear and mitochondrial genetic data to clarify the identification and distribution of leopard frog species in this region. We focus on four problematic frog populations of uncertain species affiliation in northern NJ, southeastern mainland NY, and Staten Island to test the following hypotheses: (1) they are conspecific with Rana sphenocephala or R. pipiens, (2) they are hybrids between R. sphenocephala and R. pipiens, or (3) they represent one or more previously undescribed cryptic taxa. Bayesian phylogenetic and cluster analyses revealed that the four unknown populations collectively form a novel genetic lineage, which represents a previously undescribed cryptic leopard frog species, Rana sp. nov. Statistical support for R. sp. nov. was strong in both the Bayesian (pp=1.0) and maximum-likelihood (bootstrap=99) phylogenetic analyses as well as the Structure cluster analyses. While our data support recognition of R. sp. nov. as a novel species, we recommend further study including fine-scaled sampling and ecological, behavioral, call, and morphological analyses before it is formally described.

  8. Larvicidal, Biological and Genotoxic Effects, and Temperature-Toxicity Relationship of Some Leaf Extracts of Nerium oleander (Apocynaceae) on Culex pipiens (Diptera: Culicidae)

    PubMed Central

    El-Sayed, Shaurub H; El-Bassiony, Ghada M

    2016-01-01

    Background: The present study was undertaken to study the larvicidal activity of different extracts of Nerium oleander leaves, and post-treatment temperature- toxicity relationship of these extracts against Culex pipiens. Further, the most potent extract was used to evaluate its biological and genotoxic activities. Methods: Crude extracts of N. oleander leaves were prepared using water, chloroform, acetone and diethyl ether as solvents. Extraction was carried out using soxhlet apparatus. Bioassay test was carried out on the larvae, and the LC50 of each extract was determined. Thus, newly hatched first instar larvae were treated, and the mortality count was recorded daily till pupation (accumulated mortality). The LC50 of diethyl ether extract, as the most potent extract, was used for the further biological and genotoxic studies. Results: The results obtained indicated that diethyl ether extract of N. oleander leaves was the most potent extract, with LC50 of 10500 mg/l. The toxicity of the four extracts, using the LC50, at 10 °C was higher than that at 35 °C. The LC50 of diethyl ether extract significantly decreased the larval duration, pupal duration, percentage of pupation, percentage of adult emergence, longevity of females, fecundity, and oviposition activity index, whereas the growth index and the percentage of development per day of larvae and pupae were significantly increased compared to non-treated insects. Moreover, treatment with this extract induced significant dominant lethality in both male and female adults. Conclusion: It appears that diethyl ether extract of N. oleander leaves is potential control agent to Cx. pipiens. PMID:27047967

  9. New species of Oswaldocruzia (Nematoda: Molineoidae), new species of Rhabdias (Nematoda: Rhabdiasidae), and other helminths in Rana cf. forreri (Anura: Ranidae) from Costa Rica.

    PubMed

    Bursey, Charles R; Goldberg, Stephen R

    2005-06-01

    Oswaldocruzia costaricensis n. sp. (Strongylida: Molineidae) from the intestines and Rhabdias savagei n. sp. (Rhabditida: Rhabdiasidae) from the lungs of Rana cf. forreri (Anura: Ranidae) are described and illustrated. Oswaldocruzia costaricensis represents the 77th species assigned to the genus and differs from the other Neotropical species in the genus by possessing a Type II bursa and long cervical alae. Rhabdias savagei represents the 47th species assigned to the genus and differs from other Neotropical species in the genus by possession of 4 lips and a postequatorial vulva. Rana cf. forreri was also found to harbor the trematodes, Haematoloechus parcivitellarius and Megalodiscus temperatus, the nematodes, Aplectana incerta, Aplectana itzocanensis, Cosmocerca podicipinus, Foleyellides striatus, Subulascaris falcaustriformis, and a larva of the nematode Brevimulticaecum sp. Cosmocerca panamaensis is considered to be a synonym of Cosmocerca podicipinus.

  10. Residues of polybrominated diphenyl ethers in frogs (Rana limnocharis) from a contaminated site, South China: tissue distribution, biomagnification, and maternal transfer.

    PubMed

    Wu, Jiang-Ping; Luo, Xiao-Jun; Zhang, Ying; Chen, She-Jun; Mai, Bi-Xian; Guan, Yun-Tao; Yang, Zhong-Yi

    2009-07-15

    Environmental pollutants are suspected to be a cause of global declines in amphibian populations, but few data are available on the bioaccumulation of polybrominated diphenyl ethers (PBDEs) in amphibians. To examine the tissue distribution, biomagnification potential, and maternal transfer of PBDEs in frogs, eighteen PBDE congeners were measured in the muscle, liver, and egg tissues of rice frogs (Rana limnocharis) and insects collected from an electronic waste (e-waste) recycling site in South China. PBDE levels in the frogs ranged from 0.63 to 11.6, 4.57 to 56.2, and 10.7 to 125 ng/g wet wt in the muscles, livers, and eggs, respectively. The frogs exhibited a unique congener profile, compared to those in aquatic and terrestrial species, with BDEs 99, 153, 183, 209, and 47 as the dominant congeners, intermediating between aquatic and terrestrial species. Most of the PBDE congeners in general showed higher affinity to liver than to muscle tissue. Except for BDEs 28, 47, 66, 138, and 206, the average biomagnification factors (BMFs) for all PBDE congeners were greater than 1.0, providing clear evidence of their biomagnification from insects to frogs. A parabolic relationship between log BMFs and bromine atom numbers or log Kow of PBDEs was observed, with the maximum BMF values for PBDEs with 6 bromine atoms (or at a log K(ow) of approximately 8.0). Relatively higher levels of 3-MeO-BDE 47 were found in male frogs, suggesting that male frogs in the present study might have higher metabolic capacity for PBDEs compared to female frogs. The ratio of levels in egg/female liver, indicating mother-to-egg transfer capacity, increased with increasing bromine atom numbers up to 7 and then declined as the bromine atom numbers rose. This indicated that the physicochemical properties of the congeners (e.g., K(ow), molecular sizes, and structures), resulting in different affinities to transport proteins, might impact their maternal transfer in frogs.

  11. Multiple invasions of the Ryukyu Archipelago by Oriental frogs of the subgenus Odorrana with phylogenetic reassessment of the related subgenera of the genus Rana.

    PubMed

    Matsui, Masafumi; Shimada, Tomohiko; Ota, Hidetoshi; Tanaka-Ueno, Tomoko

    2005-12-01

    The genus Rana, notably diversified in Oriental regions from China to Southeast Asia, includes a group of cascade frogs assigned to subgenera Odorrana and Eburana. Among them, R. ishikawae and the R. narina complex represent the northernmost members occurring from Taiwan to the Ryukyu Archipelago of Japan. Relationships of these frogs with the continental members, as well as the history of their invasions to islands, have been unclear. The taxonomic status of Odorrana and related genera varies among authors and no phylogenetic reassessment has been done. Using partial sequences of mitochondrial 12S and 16S rRNA genes, we estimated phylogenetic relationships among 17 species of the section Hylarana including Odorrana and Eburana, and related species from the Ryukyus, Taiwan, China, Thailand, Malaysia, and Indonesia. We estimate that (1) Odorrana is monophyletic and encompasses species of Eburana and R. hosii, which is now placed in Chalcorana, (2) the ancestor of R. ishikawae separated from other Rana in the middle to late Miocene prior to its entry to the Ryukyu Archipelago, (3) the ancestor of the R. narina complex later diversified in continental Asia, and invaded the Ryukyu Archipelago through Taiwan, (4) the R. narina complex attained its current distribution within the Ryukyus through niche segregations, and (5) vicariance of R. hosii between Malay Peninsula and Borneo occurred much later than the divergence events in the R. narina complex. Current subgeneric classification of Rana, at least of Southeast Asian members, requires full reassessment in the light of phylogenetic relationships.

  12. Cryopreservation of hormonally induced sperm for the conservation of threatened amphibians with Rana temporaria as a model research species.

    PubMed

    Shishova, N R; Uteshev, V K; Kaurova, S A; Browne, R K; Gakhova, E N

    2011-01-15

    The survival of hundreds of threatened amphibian species is increasingly dependent on conservation breeding programs (CBPs). However, there is an ongoing loss of genetic variation in CBPs for most amphibians, reptiles, birds, and mammals. Low genetic variation results in the failure of CBPs to provide genetically competent individuals for release in supplementation or rehabitation programs. In contrast, in the aquaculture of fish the perpetuation of genetic variation and the production of large numbers of genetically competent individuals for release is accomplished through the cryopreservation of sperm. Successful protocols for the cryopreservation of amphibian sperm from excised testes, and the use of motile frozen then thawed sperm for fertilisation, have been adapted from those used with fish. However, there have been no protocols published for the cryopreservation of amphibian hormonally induced sperm (HIS) that have achieved fertility. We investigated protocols for the cryopreservation of amphibian HIS with the European common frog (Rana temporaria) as a model research species. We induced spermiation in R. temporaria through the intraperitoneal administration of 50 μg LHRHa and sampled HIS through expression in spermic urine. Highly motile HIS at a concentration of 200 × 10(6)/mL was then mixed 1:1 with cryodiluents to form cryosuspensions. Initial studies showed that; 1) concentrations of ∼15 × 10(6)/mL of HIS achieve maximum fertilisation, 2) TRIS buffer in cryodiluents did not improve the recovery of sperm after cryopreservation, and 3) high concentrations of DMSO (dimethylsulphoxide) cryoprotectant reduce egg and larval survival. We then compared four optimised cryopreservation protocols for HIS with the final concentrations of cryodiluents in cryosuspensions of; 1) DMSO, (½ Ringer Solution (RS), 10% sucrose, 12% DMSO); 2) DMSO/egg yolk, (½ RS, 10% sucrose, 12% DMSO, 10% egg yolk), 3) DMFA, (½ RS, 10% sucrose, 12% dimethylformamide (DMFA)), and 4

  13. Sex-chromosome differentiation and ‘sex races’ in the common frog (Rana temporaria)

    PubMed Central

    Rodrigues, Nicolas; Vuille, Yvan; Loman, Jon; Perrin, Nicolas

    2015-01-01

    Sex-chromosome differentiation was recently shown to vary among common frog populations in Fennoscandia, suggesting a trend of increased differentiation with latitude. By rearing families from two contrasted populations (respectively, from northern and southern Sweden), we show this disparity to stem from differences in sex-determination mechanisms rather than in XY-recombination patterns. Offspring from the northern population display equal sex ratios at metamorphosis, with phenotypic sexes that correlate strongly with paternal LG2 haplotypes (the sex chromosome); accordingly, Y haplotypes are markedly differentiated, with male-specific alleles and depressed diversity testifying to their smaller effective population size. In the southern population, by contrast, a majority of juveniles present ovaries at metamorphosis; only later in development do sex ratios return to equilibrium. Even at these later stages, phenotypic sexes correlate only mildly with paternal LG2 haplotypes; accordingly, there are no recognizable Y haplotypes. These distinct patterns of gonadal development fit the concept of ‘sex races’ proposed in the 1930s, with our two populations assigned to the ‘differentiated’ and ‘semi-differentiated’ races, respectively. Our results support the suggestion that ‘sex races’ differ in the genetic versus epigenetic components of sex determination. Analysing populations from the ‘undifferentiated race’ with high-density genetic maps should help to further test this hypothesis. PMID:25833852

  14. Acid-precipitation studies in Colorado and Wyoming: Interim report of surveys of Montane amphibians and water chemistry. Interim report, 1986-1988

    SciTech Connect

    Corn, P.S.; Stolzenburg, W.; Bury, R.B.

    1989-06-01

    Surveys for amphibians were conducted in the Rocky Mountains of northern Colorado and southern Wyoming from 1986 to 1988. The northern leopard frog (Rana pipiens) was present at only 12% of historically known localities, and the boreal toad (Bufo boreas) was present at 17% of known localities. Chorus frogs (Pseudacris triseriata) suffered a catastrophic decline in population size in one population monitored since 1961, but regionally, this species was observed in 64% of known localities. Tiger salamanders (Ambystoma tigrinum) and wood frogs (Rana sylvatica) were present at 45% and 69% of known localities respectively. Acid neutralizing capacity, pH, specific conductivity, and cation concentrations in water at amphibian localities were negatively correlated with elevation. Survival of wood frog embryos declined when exposed to aluminum concentrations.

  15. Molecular structure of the prothoracicotropic hormone gene in the northern house mosquito, Culex pipiens, and its expression analysis in association with diapause and blood feeding.

    PubMed

    Zhang, Q; Denlinger, D L

    2011-04-01

    We cloned the gene that encodes prothoracicotropic hormone (PTTH) in the northern house mosquito, Culex pipiens, and investigated its expression profile in short-day (diapause-destined) and long-day (nondiapause-destined) individuals from the fourth-instar larval stage to 2 months of adulthood, as well as after a blood meal. The deduced C. pipiens PTTH (Cupip-PTTH) amino acid sequence contains seven cysteines with a specific spacing pattern. Sequence alignment suggests that Cupip-PTTH is 23% identical to Drosophila melanogaster PTTH, but is ≥59% identical to the PTTHs of other mosquitoes. Cupip-PTTH has structural characteristics similar to those of Bombyx mori PTTH and some vertebrate nerve growth factors with cysteine-knot motifs. PTTH transcripts exhibit a daily cycling profile during the final (fourth) larval instar, with peak abundance occurring late in the scotophase. The fourth-larval instar stage is one day longer in short-day larvae than in long-day larvae, resulting in larger larvae and adults. This additional day of larval development is associated with one extra PTTH cycle. No cycling was observed in pupae, but PTTH transcripts were slightly higher in short-day pupae than in long-day pupae throughout much of the pupal stage. PTTH expression persisted at a nearly constant level in diapausing adult females for the first month but then dropped by ∼50%, while expression decreased at the beginning of adulthood in nondiapausing females and then remained at a low level as long as the females were denied a blood meal. However, when nondiapausing females were offered a blood meal, PTTH transcripts rose approximately 7 fold in 2 h and remained elevated for 24 h. A few diapausing females (∼10%) will take a blood meal when placed in close proximity to a host, but much of the blood is ejected and such meals do not result in mature eggs. Yet, elevated PTTH mRNA expression was also observed in diapausing females that were force fed. Our results thus point to

  16. Nestedness in colonization-dominated systems: helminth infracommunities of Rana vaillanti Brocchi (Anura: Ranidae) in Los Tuxtlas, Veracruz, Mexico.

    PubMed

    Zelmer, Derek A; Paredes-Calderón, Laura; León-Règagnon, Virginia; García-Prieto, Luis

    2004-08-01

    Colonization probabilities of parasite species often are determined by the habitat preference and vagility of host individuals. Although extinction-based interpretations have been investigated for nested subset patterns of parasite infracommunities, the low relative frequency of nestedness in colonization-dominated systems makes the determination and interpretation of nested infracommunities of broad ecological importance. In these systems, ontogenetic shifts in habitat preference or diet of the host have the potential to produce nested subset patterns of parasite infracommunities. Helminth infracommunity structure was investigated for 76 Rana vaillanti individuals collected from Laguna Escondida, Los Tuxtlas, Veracruz, Mexico, in 1998. Pooled helminth infracommunities were significantly nested, as were penetrating and ingested helminth infracommunities when considered separately. Richness, diversity, and evenness of the helminth infracommunities were not correlated with host size, and did not differ between host sexes, suggesting that the structure of infracommunities simply is a product of the interaction between host individuals and their landscape mediated by individual differences in vagility. It is hypothesized that individual differences in recruitment can produce nested subset infracommunity patterns when the habitats or habitat preferences of hosts are themselves nested.

  17. Identification and characterization of a novel freezing inducible gene, li16, in the wood frog Rana sylvatica.

    PubMed

    McNally, J Dayre; Wu, Shao-Bo; Sturgeon, Christopher M; Storey, Kenneth B

    2002-06-01

    The wood frog Rana sylvatica survives for weeks during winter hibernation with up to 65% body water frozen as ice. Natural freeze tolerance includes both seasonal and freeze-induced molecular adaptations that control ice formation, deal with long-term ischemia, regulate cell volume changes, and protect macromolecules. This report identifies and characterizes a novel freeze-inducible gene, li16, that codes for a protein of 115 amino acids. Northern blot analysis showed that li16 transcript levels rose quickly during freezing to reach levels 3.7-fold higher than control values after 24 h; immunoblotting showed a parallel 2.4-fold rise in Li16 protein. Regulatory influences on gene expression were assessed. Nuclear runoff assays confirmed that freezing initiated an increase in the rate of li16 transcription, and analysis of signal transduction pathways via in vitro incubation of liver slices implicated a cGMP-mediated pathway in li16 expression. Gene and protein expression in liver was also strongly stimulated by anoxia exposure, whereas the gene was less responsive to dehydration stress. The strong response of li16 to both freezing and anoxia, and the rapid down-regulation of the gene when oxygen was reintroduced, suggest that the Li16 protein may play a role in ischemia resistance during freezing.

  18. Transcript expression of the freeze responsive gene fr10 in Rana sylvatica during freezing, anoxia, dehydration, and development.

    PubMed

    Sullivan, K J; Biggar, K K; Storey, K B

    2015-01-01

    Freeze tolerance is a critical winter survival strategy for the wood frog, Rana sylvatica. In response to freezing, a number of genes are upregulated to facilitate the survival response. This includes fr10, a novel freeze-responsive gene first identified in R. sylvatica. This study analyzes the transcriptional expression of fr10 in seven tissues in response to freezing, anoxia, and dehydration stress, and throughout the Gosner stages of tadpole development. Transcription of fr10 increased overall in response to 24 h of freezing, with significant increases in expression detected in testes, heart, brain, and lung when compared to control tissues. When exposed to anoxia; heart, lung, and kidney tissues experienced a significant increase, while the transcription of fr10 in response to 40% dehydration was found to significantly increase in both heart and brain tissues. An analysis of the transcription of fr10 throughout the development of the wood frog showed a relatively constant expression; with slightly lower transcription levels observed in two of the seven Gosner stages. Based on these results, it is predicted that fr10 has multiple roles depending on the needs and stresses experienced by the wood frog. It has conclusively been shown to act as a cryoprotectant, with possible additional roles in anoxia, dehydration, and development. In the future, it is hoped that further knowledge of the mechanism of action of FR10 will allow for increased stress tolerance in human cells and tissues.

  19. Environmental stress responsive expression of the gene li16 in Rana sylvatica, the freeze tolerant wood frog.

    PubMed

    Sullivan, Katrina J; Storey, Kenneth B

    2012-06-01

    Wood frogs (Rana sylvatica) can endure weeks of subzero temperature exposure during the winter with up to 65% of their body water frozen as extracellular ice. Associated with freezing survival is elevated expression of a number of genes/proteins including the unidentified gene, li16, first described in liver. The current study undertakes a broad analysis of li16 expression in response to freezing in 12 tissues of wood frogs as well as expression responses to anoxia and dehydration. Transcript levels of li16 increased significantly after 24h freezing (at -2.5 °C) demonstrating increases of approximately 3-fold in testes, greater than 2-fold in heart, ventral skin and lung, and over 1.5-fold in brain, liver and hind leg muscle as compared to unfrozen controls at 5 °C. Increased li16 transcript levels in brain, muscle and heart were mirrored by elevated Li16 protein in frozen frogs. Significant upregulation of li16 in response to both anoxia and dehydration (both components of freezing) was demonstrated in brain, kidney and heart. Overall, the results indicate that Li16 protein has a significant role to play in cell/organ responses to freezing in wood frogs and that its up-regulation may be linked with oxygen restriction that is a common element in the three stress conditions examined.

  20. Cocaine- and amphetamine-regulated transcript (CART) peptide as an in vivo regulator of cardiac function in Rana ridibunda frog.

    PubMed

    Ivanova, Iliyana V; Schubert, Rudolf; Duridanova, Dessislava B; Bolton, Thomas B; Lubomirov, Lubomir T; Gagov, Hristo S

    2007-11-01

    The aim of this study was to investigate the effect of CART peptide on cardiac performance and on the physiological signalling pathways involved using Rana ridibunda frog heart preparations in vivo. The CART peptide, when injected into the venous sinus, significantly and reproducibly increased the force of frog heart contractions by up to 33.0 +/- 6.4% during the first 15 min after its application but did not influence the chronotropic activity of the frog heart. The positive inotropic effect was entirely blocked by prazosin, pertussis toxin, R(p)-adenosine 3',5'-cyclic monophosphorothioate, autosauvagine 30 or metyrapone, as well as by extirpation of the pituitary gland, functional elimination of the inter-renal glands and long-lasting starvation, and was not observed on isolated heart preparations. Propranolol and double pithing were without significant effect on this phenomenon. It was concluded that: (i) CART peptide, administered to frogs in vivo, increases the force of heart contractions; (ii) this effect of the peptide is exerted via activation of the hypothalamic-pituitary-inter-renal gland axis through a corticoliberin-sensitive mechanism; (iii) CART augments the pumping function of the heart via a corticosteroid-dependent potentiation of myocardial alpha(1)-adrenoreceptors signalling; and (iv) prolonged food deprivation abolishes the positive inotropic effect of CART, suggesting the participation of endogenous CART in the physiological adaptation of the circulatory system to limitations of energy consumption.

  1. Ontogenic delays in effects of nitrite exposure on tiger salamanders (Ambystoma tigrinum tigrinum) and wood frogs (Rana sylvatica).

    PubMed

    Griffis-Kyle, Kerry L

    2005-06-01

    Under certain conditions, nitrite can be present in freshwater systems in quantities that are toxic to the fauna. I exposed wood frog (Rana sylvatica) and eastern tiger salamander (Ambystoma tigrinum tigrinum) embryos and young tadpoles and larvae to elevated concentrations of nitrite in chronic toxicity tests: 0, 0.3, 0.6, 1.2, 2.1, 4.6, and 6.1 mg/L NO2-N, exposing individuals as both embryos and larvae. Nitrite caused significant declines in wood frog hatching success (3.4 mg/L NO2-N, wood frog), and lower concentrations caused significant mortality during the early larval stages (4.6 mg/L NO2-N, salamander; 0.5 mg/L NO2-N, wood frog). Later tests exposing individuals to nitrite only after hatching showed that both wood frog and tiger salamander vulnerability to nitrite declined shortly after hatching. Hence, examining a single life-history stage, especially later in development, may miss critical toxic effects on organisms, causing the researcher potentially to underestimate seriously the ecological consequences of nitrite exposure.

  2. Acquisition of species-specific O-linked carbohydrate chains from oviducal mucins in Rana arvalis. A case study.

    PubMed

    Coppin, A; Maes, E; Flahaut, C; Coddeville, B; Strecker, G

    1999-12-01

    The extracellular matrix surrounding amphibian eggs is composed of mucin-type glycoproteins, highly O-glycosylated and plays an important role in the fertilization process. Oligosaccharide-alditols were released from the oviducal mucins of the anuran Rana arvalis by alkali-borohydride treatment in reduced conditions. Neutral and acidic oligosaccharides were fractionated by ion-exchange chromatographies and purified by HPLC. Each compound was identified by matrix assisted laser desorption ionization-time of flight (MALDI-TOF) spectrometry, NMR spectroscopy, electrospray ionization-tandem mass spectroscopy (ESI-MS/MS) and permethylation analyses. This paper reports on the structures of 19 oligosaccharide-alditols, 12 of which have novel structures. These structures range in size from disaccharide to octasaccharide. Some of them are acidic, containing either a glucuronic acid or, more frequently, a sulfate group, located either at the 6 position of GlcNAc or the 3 or 4 positions of Gal. This latter sulfation is novel and has only been characterized in the species R. arvalis. This structural analysis led to the establishment of several novel carbohydrate structures, demonstrating the structural diversity and species-specificity of amphibian glycoconjugates.

  3. Effects of testosterone on contractile properties of sexually dimorphic forelimb muscles in male bullfrogs (Rana catesbeiana, Shaw 1802)

    PubMed Central

    Kampe, Aaron R.; Peters, Susan E.

    2013-01-01

    Summary This study examined the effects of testosterone (T) on the contractile properties of two sexually dimorphic forelimb muscles and one non-dimorphic muscle in male bullfrogs (Rana catesbeiana, Shaw 1802). The dimorphic muscles in castrated males with testosterone replacement (T+) achieved higher forces and lower fatigability than did castrated males without replaced testosterone (T0 males), but the magnitude of the differences was low and many of the pair-wise comparisons of each muscle property were not statistically significant. However, when taken as a whole, the means of seven contractile properties varied in the directions expected of masculine values in T+ animals in the sexually dimorphic muscles. Moreover, these data, compared with previous data on male and female bullfrogs, show that values for T+ males are similar to normal males and are significantly different from females. The T0 males tended to be intermediate in character between T+ males and females, generally retaining masculine values. This suggests that the exposure of young males to T in their first breeding season produces a masculinizing effect on the sexually dimorphic muscles that is not reversed between breeding seasons when T levels are low. The relatively minor differences in contractile properties between T+ and T0 males may indicate that as circulating T levels rise during breeding season in normal males, contractile properties can be enhanced rapidly to maximal functional levels for breeding success. PMID:24143280

  4. Chloride conductance and mitochondria-rich cell density in isolated skin of Rana catesbeiana acclimated to various environments.

    PubMed

    Claro de Toledo, Manuel; Malheiros Lopes Sanioto, Sonia

    2002-08-01

    The Cl- conductance in isolated skin of frogs (Rana catesbeiana) acclimated to 30 mM solutions of NaCl, Na2SO4, MgCl2 and distilled water (DW) was studied. Transepithelial potential difference (PDtrans), short-circuit current (ISC) and total conductance (Gt) were measured under conditions such that there was Cl- flux in the presence and absence of Na+ transport. The Cl- content of the mucosal solution was acutely replaced with SO42- or gluconate to evaluate the effect of removal of Cl- conductance on electrophysiological parameters. Mitochondria-rich cell density (DMRC) was also measured. Skins from frogs acclimated to NaCl and Na2SO4 showed the lowest and the highest D(MRC), respectively, but no difference could be found between the skins from frogs acclimated to DW and MgCl2 indicating that DMRC is not unconditionally dependent on environmental Cl- in this species. Frogs acclimated to NaCl showed marked differences when compared to the other groups: the highest Gt, probably represented by a higher paracellular conductance; the lowest transepithelial electrical potential difference which remained invariant after replacement of mucosal Cl- with SO42- or replacement of mucosal Cl- with gluconate and an inwardly oriented positive current in the absence of bilateral Na+.

  5. Effects of octylphenol on the expression of StAR, CYP17 and CYP19 in testis of Rana chensinensis.

    PubMed

    Bai, Yao; Li, Xin-Yi; Liu, Zhi-Jun; Zhang, Yu-Hui

    2017-02-21

    It has been proposed that a decline in sperm quality is associated with exposure to environmental chemicals with estrogenic activity. Seeking possible explanations for this effect, this study investigated the effects of octylphenol (OP) on the synthesis of steroid hormones in amphibian. Rana chensinensis were exposed to 10(-8), 10(-7) and 10(-6)mol/L OP after 10, 20, 30 and 40 days. The cDNA fragments of StAR (274bp), CYP17 (303bp) and CYP19 (322bp) were cloned. In situ hybridization and immunohistochemistry revealed that positive signals of StAR, CYP17, CYP19 mRNA and proteins mainly in the Leydig cells of testes. Real-time PCR showed that up-regulation of StAR and CYP19, and down-regulation of CYP17 after exposure to 10(-8), 10(-7) and 10(-6)mol/L OP. The results suggest that OP can alter transcriptions of StAR, CYP17 and CYP19, thus disturb the expressions of StAR, P450c17 and P450arom, thereby adversely affect steroid synthesis.

  6. Different responses of biochemical markers in frogs (Rana ridibunda) from urban and rural wetlands to the effect of carbamate fungicide.

    PubMed

    Falfushinska, Halina I; Romanchuk, Liliya D; Stolyar, Oksana B

    2008-09-01

    Laboratory studies were conducted to determine the effects of carbamate fungicide TATTU (mixture of propamocarb and mancozeb, 0.091 mg L(-1)) on biochemical markers of exposure in Rana ridibunda from clean (reference) and polluted sites. The untreated animals from the polluted site had lower Cu,Zn- and Mn-superoxide dismutase (SOD) and acetylcholinesterase activity, the levels of lipid peroxidation products (TBARS) and protein carbonyls in the liver and vitellogenin-like proteins (Vtg-LP) in the serum, but higher levels of glutathione in the liver in comparison with untreated frogs from the reference site. Catalase activity, superoxide anion and metallothionein levels were the same in both groups. The animals from two sites demonstrate different response on the effect of TATTU during 14 days. In the frogs from polluted site the oxidative damage (the decrease of Mn-SOD activity, lipids and protein oxidative destruction), neurotoxicity (depletion of acetylcholinesterase activity), and endocrine disruption (increase of Vtg-LP level) were revealed. On the other hand, the part of the indices in the animals from the reference site was unchanged after the treatment and the level of metallothionein was elevated demonstrating the satisfactory ability for the adaptation to unfavourable conditions.

  7. Metabolic depression induced by urea in organs of the wood frog, Rana sylvatica: effects of season and temperature.

    PubMed

    Muir, Timothy J; Costanzo, Jon P; Lee, Richard E

    2008-03-01

    It has long been suspected that urea accumulation plays a key role in the induction or maintenance of metabolic suppression during extended dormancy in animals from diverse taxa. However, little evidence supporting that hypothesis in living systems exists. We measured aerobic metabolism of isolated organs from the wood frog (Rana sylvatica) in the presence or absence of elevated urea at various temperatures using frogs acclimatized to different seasons. The depressive effect of urea on metabolism was not consistent across organs, seasons, or temperatures. None of the organs from summer frogs, which were tested at 20 degrees C, or from winter frogs tested at 4 degrees C were affected by urea treatment. However, liver, stomach, and heart from spring frogs tested at 4 degrees C had significantly lower metabolic rates when treated with urea as compared with control samples. Additionally, when organs from winter frogs were tested at 10 degrees C, metabolism was significantly decreased in urea-treated liver and stomach by approximately 15% and in urea-treated skeletal muscle by approximately 50%. Our results suggest that the presence of urea depresses the metabolism of living organs, and thereby reduces energy expenditure, but its effect varies with temperature and seasonal acclimatization. The impact of our findings may be wide ranging owing to the number of diverse organisms that accumulate urea during dormancy.

  8. Toxic effects of octylphenol on the expression of genes in liver identified by suppression subtractive hybridization of Rana chensinensis.

    PubMed

    Li, Xin-Yi; Xiao, Ning; Zhang, Yu-Hui

    2014-01-01

    Octylphenol (OP) is the degradative product of alkylphenol ethoxylates that are widely used to produce rubber, pesticides, and paints. It is chemically stable substance and demonstrates estrogenic effects, toxicity and carcinogenic effects in the environment. The toxin accumulates rapidly in the liver where it exerts most of its damage, but the molecular mechanisms behind its toxicity remain unclear. Due to limited information concerning the effect of OP on liver, this study investigates how OP causes hepatotoxicity in liver. Here, suppression subtractive hybridization was used to identify the alterations in gene transcription of the frog (Rana chensinensis) after exposure to OP. After hybridization and cloning, the subtractive cDNA libraries were obtained. At random, 207 positive clones were selected and sequenced from the subtractive libraries, which gave a total of 75 gene fragment sequences. The screening identified numerous genes involved in apoptosis, signal transduction, cytoskeletal remodeling, innate immunity, material and energy metabolism, translation and transcription which were extensively discussed. Two sequenced genes were analyzed further using real time quantitative PCR. The two genes from the library were found to be transcriptionally up-regulated. These results confirmed the successful construction of the subtractive cDNA library that was enriched for the genes that were differentially transcribed in the amphibian liver challenged with OP, and for the first time present the basic data on toxicity effect of OP on liver.

  9. Odorous and Non-Fatal Skin Secretion of Adult Wrinkled Frog (Rana rugosa) Is Effective in Avoiding Predation by Snakes

    PubMed Central

    Yoshimura, Yuri; Kasuya, Eiiti

    2013-01-01

    The roles played by nonfatal secretions of adult anurans in the avoidance of predation remain unknown. The adult Wrinkled frog (Rana rugosa) has warty skin with the odorous mucus secretion that is not fatal to the snake Elaphe quadrivirgata. We fed R. rugosa or Fejervarya limnocharis, which resembles R. rugosa in appearance and has mucus secretion, to snakes and compared the snakes’ responses to the frogs. Compared to F. limnocharis, R. rugosa was less frequently bitten or swallowed by snakes. The snakes that bit R. rugosa spat out the frogs and showed mouth opening (gaping) behavior, while the snakes that bit F. limnocharis did not show gaping behavior. We also compared the responses of the snakes to R. rugosa and F. limnocharis secretions. We coated palatable R. japonica with secretions from R. rugosa or F. limnocharis. The frogs coated by R. rugosa secretion were less frequently bitten or swallowed than those coated by F. limnocharis secretion. We concluded that compared to different frog species of similar sizes, the adult R. rugosa was less frequently preyed upon by, and that its skin secretion was effective in avoiding predation by snakes. PMID:24278410

  10. Odorous and non-fatal skin secretion of adult wrinkled frog (Rana rugosa) is effective in avoiding predation by snakes.

    PubMed

    Yoshimura, Yuri; Kasuya, Eiiti

    2013-01-01

    The roles played by nonfatal secretions of adult anurans in the avoidance of predation remain unknown. The adult Wrinkled frog (Rana rugosa) has warty skin with the odorous mucus secretion that is not fatal to the snake Elaphe quadrivirgata. We fed R. rugosa or Fejervarya limnocharis, which resembles R. rugosa in appearance and has mucus secretion, to snakes and compared the snakes' responses to the frogs. Compared to F. limnocharis, R. rugosa was less frequently bitten or swallowed by snakes. The snakes that bit R. rugosa spat out the frogs and showed mouth opening (gaping) behavior, while the snakes that bit F. limnocharis did not show gaping behavior. We also compared the responses of the snakes to R. rugosa and F. limnocharis secretions. We coated palatable R. japonica with secretions from R. rugosa or F. limnocharis. The frogs coated by R. rugosa secretion were less frequently bitten or swallowed than those coated by F. limnocharis secretion. We concluded that compared to different frog species of similar sizes, the adult R. rugosa was less frequently preyed upon by, and that its skin secretion was effective in avoiding predation by snakes.

  11. Changes in formaldehyde-induced fluorescence of the hypothalamus and pars intermedia in the frog, Rana temporaria, following background adaptation.

    PubMed

    Prasada Rao, P D

    1982-01-01

    Adaptation of the frog, Rana temporaria, to a white background for 12 hr has resulted in an intense formaldehyde-induced fluorescence (FIF) in the neurons of the preoptic recess organ (PRO), paraventricular organ (PVO), nucleus infundibularis dorsalis (NID) and their basal processes permitting visualization of the PRO- and PVO-hypophysial tracts that extend into the median eminence (ME) and pars intermedia (PI); the FIF is reduced in all the structures by 3 days. In frogs adapted to a black background, for 12 hr and 3 days, there was a general reduction in the FIF of the PRO neurons and PRO-hypophysial tract. By 12 hr black background adaptation, the PVO/NID neurons and only their adjacent basal processes show FIF which was sharply reduced by 3 days, making the PVO-hypophysial tract undetectable. In the PI fibers the fluorescence was more intense in black-adapted frogs than in white-adapted ones at both the intervals studied. The simultaneous changes in the FIF of the hypothalamic nuclei, tracts and PI suggest that the PRO and PVO/NID neurons participate in PI control through release of neurotransmitter(s) at the axonal ends.

  12. Upregulation of two actin genes and redistribution of actin during diapause and cold stress in the northern house mosquito, Culex pipiens.

    PubMed Central

    Kim, Mijung; Robich, Rebecca M.; Rinehart, Joseph P.; Denlinger, David L.

    2007-01-01

    Two actin genes cloned from Culex pipiens L. are upregulated during adult diapause. Though actins 1 and 2 were expressed throughout diapause, both genes were most highly expressed early in diapause. These changes in gene expression were accompanied by a conspicuous redistribution of polymerized actin that was most pronounced in the midguts of diapausing mosquitoes that were exposed to low temperature. In nondiapausing mosquitoes reared at 25°C and in diapausing mosquitoes reared at 18°C, polymerized actin was clustered at high concentrations at the intersections of the muscle fibers that form the midgut musculature. When adults 7–10 days post-eclosion were exposed to low temperature (-5°C for 12h), the polymerized actin was evenly distributed along the muscle fibers in both nondiapausing and diapausing mosquitoes. Exposure of older adults (1month post-eclosion) to low temperature (−5°C for 12h) elicited an even greater distribution of polymerized actin, an effect that was especially pronounced in diapausing mosquitoes. These changes in gene expression and actin distribution suggest a role for actins in enhancing survival of diapausing adults during the low temperatures of winter by fortification of the cytoskeleton. PMID:17078965

  13. Identification and localization of gastrointestinal hormones in the skin of the bullfrog Rana catesbeiana during periods of activity and hibernation.

    PubMed

    Wang, Huan; Zhou, Naizhen; Zhang, Rui; Wu, Yuanyuan; Zhang, Ruidong; Zhang, Shengzhou

    2014-10-01

    Amphibian skin and its secretions contain a wide variety of biogenic amines and biologically active peptides, some of which are either identical or highly homologous to gastrointestinal hormones (GHs) of higher vertebrates. This study investigated the distribution density and immunoreactive (IR) intensity of 5-hydroxytryptamine (5-HT), gastrin (GAS), somatostatin (SS), pancreatic polypeptide (PP), neuropeptide Y (NPY) and glucagon (GLU) IR cells in the skin of the bullfrog Rana catesbeiana during periods of activity and hibernation. The results indicated that the six types of GHs were all present in the bullfrog skin and were most predominant in the epidermis and mucous glands. In dorsal skin, the density of the GHs-IR cells in mucous glands was higher than that in epidermis except for GAS-IR cells. In ventral skin, the density of 5-HT, PP and NPY-IR cells in mucous glands was also higher than that in the epidermis. During hibernation, the density of the six types of GHs-IR cells and the IR intensity of GAS, SS, NPY and GLU-IR cells in the epidermis of dorsal skin increased significantly. The IR intensity of SS, PP and NPY-IR cells in granular glands of ventral skin also increased significantly during hibernation. These results suggested that multiple types of GHs-IR cells present in the skin of R. catesbeiana, may play important roles in the regulation of the physiological functions of skin. Also, adaptive changes in the density and IR intensity of GHs-IR cells occurred during hibernation.

  14. Effects of feeding on metabolism, gas transport, and acid-base balance in the bullfrog Rana catesbeiana.

    PubMed

    Busk, M; Jensen, F B; Wang, T

    2000-01-01

    Massive feeding in ectothermic vertebrates causes changes in metabolism and acid-base and respiratory parameters. Most investigations have focused on only one aspect of these complex changes, and different species have been used, making comparison among studies difficult. The purpose of the present study was, therefore, to provide an integrative study of the multiple physiological changes taking place after feeding. Bullfrogs (Rana catesbeiana) partly submerged in water were fed meals (mice or rats) amounting to approximately (1)/(10) of their body weight. Oxygen consumption increased and peaked at a value three times the predigestive level 72-96 h after feeding. Arterial PO(2) decreased slightly during digestion, whereas hemoglobin-bound oxygen saturation was unaffected. Yet, arterial blood oxygen content was pronouncedly elevated because of a 60% increase in hematocrit, which appeared mediated via release of red blood cells from the spleen. Gastric acid secretion was associated with a 60% increase in plasma HCO3(-) concentration ([HCO3(-)]) 48 h after feeding. Arterial pH only increased from 7.86 to 7.94, because the metabolic alkalosis was countered by an increase in PCO(2) from 10.8 to 13.7 mm Hg. Feeding also induced a small intracellular alkalosis in the sartorius muscle. Arterial pH and HCO3(-) returned to control values 96-120 h after feeding. There was no sign of anaerobic energy production during digestion as plasma and tissue lactate levels remained low and intracellular ATP concentration stayed high. However, phosphocreatine was reduced in the sartorius muscle and ventricle 48 h after feeding.

  15. Characterization of the binding of [3H]CGP54626 to GABAB receptors in the male bullfrog (Rana catesbeiana).

    PubMed

    Asay, Matthew J; Boyd, Sunny K

    2006-06-13

    Gamma-aminobutyric acid (GABA) is the main inhibitory neurotransmitter in the vertebrate brain. GABA activates both ionotropic (GABA(A)) and metabotropic (GABA(B)) receptors in mammals. Whether non-mammalian vertebrates possess receptors with similar characteristics is not well understood. We used a mammalian GABA(B)-specific antagonist to determine the pharmacology of putative receptors in the brain of an anuran amphibian, the male bullfrog (Rana catesbeiana). Receptor binding assays with the antagonist [(3)H]CGP54626 revealed a single class of high affinity binding sites (with a K(D) of 2.97 nM and a B(max) of 2619 fmol/mg protein). Binding was time- and temperature-dependent, saturable and specific. Specific binding of [(3)H]CGP54626 was inhibited by several mammalian GABA(B) receptor agonists and antagonists. The rank order potency of agonists was: GABA = SKF97541 > (R)-Baclofen > 3-APPA. The rank order for antagonists was: CGP54626 = CGP55845 > CGP52432 > CGP35348. The GABA(A) receptor ligands muscimol and SR95531 had very low affinity for [(3)H]CGP54626 binding sites, while bicuculline compounds had no affinity. Binding of GABA was positively modulated by CGP7930. Taurine did not allosterically modulate GABA binding but did inhibit [(3)H]CGP54626 binding in a linear fashion. Bullfrog brain thus possesses binding sites with significant similarity to mammalian GABA(B) receptors. These receptors differ from mammalian receptors, however, in dissociation kinetics, ligand specificity and allosteric modulation.

  16. Amelioration of radiation-induced skin injury by HIV-TAT-mediated protein transduction of RP-1 from Rana pleurade.

    PubMed

    Zhang, Shuyu; Wang, Wenjie; Peng, Ying; Gu, Qing; Luo, Judong; Zhou, Jundong; Wu, Jinchang; Hou, Yinglong; Cao, Jianping

    2014-01-01

    Radiation-induced reactive oxygen species (ROS) can damage DNA and most other biological macromolecules in skin and radiation-induced skin injury is a serious concern for radiation therapy. Skin possesses an extremely efficient antioxidant system, which is conferred by two systems: antioxidant enzymes and small molecules that can scavenge ROS by donating electrons. Amphibian skin is a multifunctional organ, which protects against dangers of various oxidative stresses. Recently, a small peptide called RP-1 was isolated from the skin secretions of Rana pleurade, which shows strong antioxidant activity. However, this RP-1 peptide is limited because its inability to across the cell membrane. Protein transduction domains (PTDs) have demonstrated high efficiency for facilitating the internalization of both homologous and heterogeneous proteins into cells. This study aims to elucidate the protective effects of a HIV-TAT (TAT) PTD-coupled RP-1 fusion protein (TAT-RP1) on radiation-induced skin injury in vitro and in vivo. The synthesized fusion TAT-RP1 peptide can be incorporated into human keratinocyte HaCaT cells in a dose- and time-dependent manner without cytotoxicity. We then evaluated the protective role of TAT-RP1 against ionizing radiation. TAT-RP1 supplementation increased anti-superoxide anion ability of HaCaT cells and decreased HaCaT cell radiosensitivity to irradiation. Moreover, TAT-RP1 was able to penetrate the skin of rats, entering epidermis as well as the dermis of the subcutaneous layer in skin tissue. Topical spread of TAT-RP1 promoted the amelioration of radiation-induced skin damage in rats. These results suggest that TAT-RP1 has potential as a protein therapy for radiation-induced skin injury.

  17. Chilled frogs are hot: hibernation and reproduction of the Endangered mountain yellow-legged frog Rana muscosa

    USGS Publications Warehouse

    Santana, Frank E.; Swaisgood, Ronald R.; Lemm, Jeffrey M.; Fisher, Robert N.; Clark, Rulon W.

    2015-01-01

    In the face of the sixth great extinction crisis, it is imperative to establish effective breeding protocols for amphibian conservation breeding programs. Captive efforts should not proceed by trial and error, nor should they jump prematurely to assisted reproduction techniques, which can be invasive, difficult, costly, and, at times, counterproductive. Instead, conservation practitioners should first look to nature for guidance, and replicate key conditions found in nature in the captive environment, according to the ecological and behavioral requirements of the species. We tested the effect of a natural hibernation regime on reproductive behaviors and body condition in the Endangered mountain yellow-legged frog Rana muscosa. Hibernation had a clear positive effect on reproductive behavior, manifesting in vocal advertisement signaling, female receptivity, amplexus, and oviposition. These behaviors are critical components of courtship that lead to successful reproduction. Our main finding was that captive R. muscosa require a hibernation period for successful reproduction, as only hibernated females produced eggs and only hibernated males successfully fertilized eggs. Although hibernation also resulted in a reduced body condition, the reduction appeared to be minimal with no associated mortality. The importance of hibernation for reproduction is not surprising, since it is a major component of the conditions that R. muscosa experiences in the wild. Other amphibian conservation breeding programs can also benefit from a scientific approach that tests the effect of natural ecological conditions on reproduction. This will ensure that captive colonies maximize their role in providing genetic reservoirs for assurance and reintroduction efforts.

  18. Physiological features of the opercularis muscle and their effects on vibration sensitivity in the bullfrog Rana catesbeiana.

    PubMed

    Hetherington, T E

    1987-09-01

    The amphibian opercularis muscle connects a movable otic element (the operculum) to the pectoral girdle and can act in reception of ground vibrations. Various physiological parameters of the opercularis muscle of the bullfrog Rana catesbeiana were measured and compared with similar measurements on the iliofibularis muscle of the hindlimb. The opercularis muscle is a very slowly contracting muscle, with a Vmax of 1.81 muscle lengths s-1 compared to a Vmax of 6.24 muscle lengths s-1 for the iliofibularis muscle. The opercularis muscle develops tension slowly, taking about 10 s to attain maximum isometric tension when stimulated at 100 Hz. The muscle can retain high levels of tension for several minutes, and following stimulation has a time to half-relaxation of about 4-6 s. The slow velocity of contraction, slow rate of tension development, fatigue-resistance and slow rate of relaxation of the opercularis muscle support morphological evidence that it consists mostly of tonic muscle fibres. Experiments were also made to examine the effects of muscle tension on reception of ground vibrations as measured by inner ear microphonics. Severing the nerve supplying the opercularis muscle produced slight decreases of no more than 2 dB in responses to vibrations from 25 to 200 Hz. Artificial stimulation of the opercularis muscle after severing the nerve supplying the muscle increased responses to vibration across the entire frequency range. Higher tension levels produced greater increases in responses; at the highest tensions used (about 120 kN m-2) responses were increased by as much as 4.5 dB. The opercularis muscle is therefore specialized for slow but prolonged contractions, and tension is important in its sensory function. A tensed opercularis muscle appears to transmit faithfully motion of the forelimb, produced by vibrations, to the operculum such that the latter moves relative to the inner ear fluids.

  19. Anti-apoptotic response during anoxia and recovery in a freeze-tolerant wood frog (Rana sylvatica)

    PubMed Central

    Gerber, Victoria E.M.; Wijenayake, Sanoji

    2016-01-01

    The common wood frog, Rana sylvatica, utilizes freeze tolerance as a means of winter survival. Concealed beneath a layer of leaf litter and blanketed by snow, these frogs withstand subzero temperatures by allowing approximately 65–70% of total body water to freeze. Freezing is generally considered to be an ischemic event in which the blood oxygen supply is impeded and may lead to low levels of ATP production and exposure to oxidative stress. Therefore, it is as important to selectively upregulate cytoprotective mechanisms such as the heat shock protein (HSP) response and expression of antioxidants as it is to shut down majority of ATP consuming processes in the cell. The objective of this study was to investigate another probable cytoprotective mechanism, anti-apoptosis during oxygen deprivation and recovery in the anoxia tolerant wood frog. In particular, relative protein expression levels of two important apoptotic regulator proteins, Bax and p-p53 (S46), and five anti-apoptotic/pro-survival proteins, Bcl-2, p-Bcl-2 (S70), Bcl-xL, x-IAP, and c-IAP in response to normoxic, 24 Hr anoxic exposure, and 4 Hr recovery stages were assessed in the liver and skeletal muscle using western immunoblotting. The results suggest a tissue-specific regulation of the anti-apoptotic pathway in the wood frog, where both liver and skeletal muscle shows an overall decrease in apoptosis and an increase in cell survival. This type of cytoprotective mechanism could be aimed at preserving the existing cellular components during long-term anoxia and oxygen recovery phases in the wood frog. PMID:27042393

  20. Discordance between mitochondrial DNA genealogy and nuclear DNA genetic structure in the two morphotypes of Rana tagoi tagoi (Amphibia: Anura: Ranidae) in the Kinki Region, Japan.

    PubMed

    Eto, Koshiro; Matsui, Masafumi; Sugahara, Takahiro

    2013-07-01

    Two morphotypes, with a large and small body size, of a brown frog Rana t. tagoi occur sympatrically in the Kinki region, central Honshu of Japan. Previous mitochondrial (mt) DNA genealogical study recognized two main lineages (A and B) and several sublineages in R. tagoi, where the small type was placed in the group A-1b, and the large type in groups A-1a and B-2a. Using haplotype network and structure analysis of three nuclear genes, we examined the discrepancy between morphology and mitochondrial genealogy. The results showed that the small type is reproductively isolated from its co-occurring large type (A-1a or B-2a), and that unlimited gene flow occurred between parapatrically occurring two mtDNA lineages of large types (A-1a and B-2a). Discordant genetic relationships between mtDNA and nuclear DNA results may be caused by the past mitochondrial introgression, and possibly, the incomplete lineage sorting. These results also suggest a heterospecific relationship between the large (A-1a and B-2a) and small types (A-1b). The large type is identified as Rana t. tagoi as it is genetically very close to the topotypes of the nominal subspecies, while the small type remains unnamed.

  1. Predation and control efficacies of Misgurnus mizolepis (Cypriniformes: Cobitidae) toward Culex pipiens molestus (Diptera: Culicidae) and fish toxicity of temephos in laboratory and septic tank conditions.

    PubMed

    Chae, Seong Chun; Kwon, Young Hyun; Min, Kyung Il; Kim, Hyung Soo; Kim, Nam-Jin; Kim, Jun-Ran; Son, Bong Gi; Ahn, Young-Joon

    2014-07-01

    Culex pipiens molestus Forskal (Diptera: Culicidae) is the dominant mosquito species in septic tanks in South Korea. An assessment was made of the biological control potential of mud loaches, Misgurnus mizolepis Günther (Cypriniformes: Cobitidae), toward Cx. p. molestus larvae in laboratory and septic tanks. Results were compared with those of temephos 20% emulsifiable concentrate. In laboratory tests, all mud loaches survived on sedimentation chamber- and effluent chamber-collected water of aerobic septic tanks (ASTs), whereas all mud loaches died within 3-12 h after introduction into sedimentation chamber- and effluent chamber-collected water of anaerobic septic tanks, Gill hyperplasia and hemorrhages at the bases of pectoral fins were detected in all dead mud loaches. These appeared to have been caused by bacterial disease, rather than the physical and chemical characteristics of the septic tank water. A mud loach consumed an average range of 1,072-1,058 larvae of Cx. p. molestus in the AST water at 24 h. At the manufacturer's recommended rate (10 ml/ton) in the AST water, the temephos formulation did not cause fish mortality. In the AST experiment, predation of mosquito larvae by mud loaches at a release rate of one fish per 900 mosquito larvae resulted in complete mosquito control from the third day after treatment throughout the 18-wk survey period, compared with temephos 20% emulsifiable concentrate-treated AST water (reduction rate, 40% at 28 days after treatment). Reasonable mosquito control in aerobic septic tanks can be achieved by mosquito breeding season stocking of a rate of one mud loach per 900 mosquito larvae.

  2. Stable coexistence of incompatible Wolbachia along a narrow contact zone in mosquito field populations.

    PubMed

    Atyame, Célestine M; Labbé, Pierrick; Rousset, François; Beji, Marwa; Makoundou, Patrick; Duron, Olivier; Dumas, Emilie; Pasteur, Nicole; Bouattour, Ali; Fort, Philippe; Weill, Mylène

    2015-01-01

    In arthropods, the intracellular bacteria Wolbachia often induce cytoplasmic incompatibility (CI) between sperm and egg, which causes conditional embryonic death and promotes the spatial spread of Wolbachia infections into host populations. The ability of Wolbachia to spread in natural populations through CI has attracted attention for using these bacteria in vector-borne disease control. The dynamics of incompatible Wolbachia infections have been deeply investigated theoretically, whereas in natural populations, there are only few examples described, especially among incompatible infected hosts. Here, we have surveyed the distribution of two molecular Wolbachia strains (wPip11 and wPip31) infecting the mosquito Culex pipiens in Tunisia. We delineated a clear spatial structure of both infections, with a sharp contact zone separating their distribution areas. Crossing experiments with isofemale lines from different localities showed three crossing types: wPip11-infected males always sterilize wPip31-infected females; however, while most wPip31-infected males were compatible with wPip11-infected females, a few completely sterilize them. The wPip11 strain was thus expected to spread, but temporal dynamics over 7 years of monitoring shows the stability of the contact zone. We examined which factors may contribute to the observed stability, both theoretically and empirically. Population cage experiments, field samples and modelling did not support significant impacts of local adaptation or assortative mating on the stability of wPip infection structure. By contrast, low dispersal probability and metapopulation dynamics in the host Cx. pipiens probably play major roles. This study highlights the need of understanding CI dynamics in natural populations to design effective and sustainable Wolbachia-based control strategies.

  3. Effects of road de-icing salt (NaCl) on larval wood frogs (Rana sylvatica).

    PubMed

    Sanzo, Domenico; Hecnar, Stephen J

    2006-03-01

    Vast networks of roads cover the earth and have numerous environmental effects including pollution. A major component of road runoff in northern countries is salt (mostly NaCl) used as a winter de-icing agent, but few studies of effects of road salts on aquatic organisms exist. Amphibians require aquatic habitats and chemical pollution is implicated as a major factor in global population declines. We exposed wood frog tadpoles to NaCl. Tests revealed 96-h LC50 values of 2,636 and 5,109 mg/l and tadpoles experienced reduced activity, weight, and displayed physical abnormalities. A 90 d chronic experiment revealed significantly lower survivorship, decreased time to metamorphosis, reduced weight and activity, and increased physical abnormalities with increasing salt concentration (0.00, 0.39, 77.50, 1,030.00 mg/l). Road salts had toxic effects on larvae at environmentally realistic concentrations with potentially far-ranging ecological impacts. More studies on the effects of road salts are warranted.

  4. Nitric oxide changes its role as a modulator of respiratory motor activity during development in the bullfrog (Rana catesbeiana).

    PubMed

    Hedrick, Michael S; Chen, Anna K; Jessop, Kristy L

    2005-10-01

    Nitric oxide (NO) is a unique chemical messenger that has been shown to play a role in the modulation of breathing in amphibians and other vertebrates. In the post-metamorphic tadpole and adult amphibian brainstem, NO, acting via the neuronal isoform of nitric oxide synthase (nNOS), is excitatory to the generation of lung burst activity. In this study, we examine the modulation of breathing by NO during development of the amphibian brainstem. Isolated brainstem preparations from pre-metamorphic and late-stage post-metamorphic tadpoles (Rana catesbeiana) were used to determine the role of NO in modulating central respiratory neural activity. Respiratory neural activity was monitored with suction electrodes recording extracellular activity of cranial nerve rootlets that innervate respiratory musculature. Brainstems were superfused with an artificial cerebrospinal fluid (aCSF) at 20-22 degrees C containing l-nitroarginine (l-NA; 1-10 mM), a non-selective NOS inhibitor. In pre-metamorphic tadpoles, l-NA increased fictive gill ventilation frequency and amplitude, and increased lung burst frequency. By contrast, l-NA applied to the post-metamorphic tadpole brainstem had little effect on fictive buccal activity, but significantly decreased lung burst frequency and the frequency of lung burst episodes. These data indicate that early in development, NO provides a tonic inhibitory input to gill and lung burst activity, but as development progresses, NO provides an excitatory input to lung ventilation. This changing role for NO coincides with the shift in importance in the different respiratory modes during development in amphibians; that is, pre-metamorphic tadpoles rely predominantly on gill ventilation whereas post-metamorphic tadpoles have lost the gills and are obligate air-breathers primarily using lungs for gas exchange. We hypothesize that NO provides a tonic input to the respiratory CPG during development and this changing role reflects the modulatory influence of NO

  5. Experimental infections of Rana esculenta with Aeromonas hydrophila: a molecular mechanism for the control of the normal flora.

    PubMed

    Simmaco, M; Mangoni, M L; Boman, A; Barra, D; Boman, H G

    1998-10-01

    Frogs can be useful models for studying the mechanisms that may regulate their natural microbial flora. Their skin glands produce a secretion containing 20-30 different peptides, some antimicrobial some neurotrophic. As they often live in soil or silt that is rich in microbes, they can be expected to be able to prevent or eliminate infections in very short periods of time. The bacterium Aeromonas hydrophila is widely distributed in nature and is considered as part of the natural flora of frogs and many animals, including humans. From an alternative frog strain of A. hydrophila, Bo-3, we isolated a spontaneous and stable mutant (Bo-3N), resistant to nalidixic acid, here used to follow the host-microbe interactions in experimental infection of mouth and skin of Rana esculenta. The skin peptides had been previously isolated, sequenced and cloned. We showed that skin treatment with a glucocorticoid (GC) cream blocked de novo synthesis of these peptides and, simultaneously, prepropeptide mRNAs disappeared while IkappaBalpha was up-regulated. Experimental mouth infections with 20 million cells of A. hydrophila Bo-3N showed that a normal wild frog can eliminate the bacteria from the mouth within 15 min, while a frog pretreated with GC cream for 1 h could not reduce Bo-3N below 3500 colony-forming units (CFU)/5 microl 'saliva'. An in vitro comparison showed that frog blood or serum allowed bacteria to grow, while the skin secretion killed the bacteria within 10 min. Using different enzyme-linked immunosorbent assays (ELISAs) with rabbit anti-Bo-3 serum as a positive control, we were able to rule out immunoglobulin G (IgG) binding to A. hydrophila. An assay for immunoglobulin M (IgM) (or some other serum component) in frog serum showed binding to A. hydrophila only corresponding to a few per cent of the positive control. For skin infections we bathed the frogs for 10 min in an overnight culture of Bo-3N diluted to about 10(7) CFU/ml. Electrical stimulation after the bath

  6. Regulation of the respiratory central pattern generator by chloride-dependent inhibition during development in the bullfrog (Rana catesbeiana).

    PubMed

    Broch, Lise; Morales, Rey D; Sandoval, Anthony V; Hedrick, Michael S

    2002-04-01

    Isolated brainstem preparations from larval (tadpole) and adult Rana catesbeiana were used to examine inhibitory mechanisms for developmental regulation of the respiratory central pattern generator (CPG). Preparations were superfused at 20-22 degrees C with Cl(-)-free artificial cerebrospinal fluid (aCSF) or with aCSF containing agonists/antagonists of gamma-aminobutyric acid (GABA) or glycine receptors. Respiratory motor output from the CPG, measured as neural activity from cranial nerve roots, was associated with fictive gill ventilation and lung ventilation in tadpoles and with fictive lung ventilation in adults. In tadpoles, fictive lung burst frequency was 0.8+/-0.2 min(-1) and did not change significantly with Cl(-)-free aCSF superfusion; however, lung burst amplitude increased by nearly 400 % (P<0.01). Fictive gill ventilation averaged 41.6+/-3.3 min(-1) and was reversibly abolished by Cl(-)-free aCSF. Superfusion with Cl(-)-free aCSF abolished lung bursts in two of seven adult preparations, and overall lung burst frequency decreased from 3.1+/-0.7 to 0.4+/-0.03 min(-1) (P<0.01), but burst amplitude was unchanged. Low concentrations of GABA (0.5 mmol l(-1)) produced a significant increase in lung burst frequency followed by almost complete inhibition at 5.0 mmol l(-1), accompanied by the abolition of gill ventilation at 2.5-5.0 mmol l(-1). By contrast, fictive lung ventilation in adults was inhibited in a dose-dependent manner by glycine and GABA, and inhibition occurred at approximately 10-fold lower concentrations compared with tadpoles. The glycine receptor antagonist strychnine (2.5-25.0 micromol l(-1)) and the GABA(A) receptor antagonist bicuculline (1-10 micromol l(-1)) inhibited fictive gill ventilation and increased fictive lung ventilation in tadpoles. However, bicuculline and strychnine inhibited fictive lung ventilation in adults. These results suggest that lung ventilation in the tadpole brainstem may be driven by a pacemaker-like mechanism since

  7. [Ploidy and genetic structure of hybrid populations of water frogs Pelophylax esculentus (L., 1758) complex (Amphibia, Ranidae) of Ukraine].

    PubMed

    Mezhzherin, S V; Morozov-Leonov, S Iu; Rostovskaia, O V; Shabanov, D A; Sobolenko, L Iu

    2010-01-01

    The present study of green frog hybrid populations of Ukraine, including analysis of allozyme variability and planimetric analysis oferythrocytes size has confirmed that the unique region in this area is the Severski Donets basin The allopolyploid individuals there are met very frequently (5.7% of all investigated frogs). In other areas of Ukraine only two polyploid hybrids have been recorded. Beside that, one frog was defined as triploid Rana ridibundus. According to our investigations, all triploid hybrids from the Severski Donets basin are identified as P. esculentu (=lessonae)--2 ridibundus males.

  8. [Characteristics of the phase-dependent vagus effects in the heart of the frog Rana temporaria and of the cod Gadus morhua].

    PubMed

    Kopylova, G N; Sokolova, N A; Samonina, G E; Krupnova, E P

    1987-01-01

    In experiments on the heart of the cod Gadus morhua and frog Rana temporaria in situ, studies have been made of changes in the heart rate induced by stimulation of the vagal nerve by single brief bursts delivered at various intervals after P wave of the ECG. Certain differences were found in changes of the heart rate between these animals. In the cod, maximum chronotropic effect was equal to 65% of the duration of initial cardiac cycle, the latency of this effect being equal to 290 ms; in the frog, corresponding figures were 12-13% and approximately 940 ms. The duration of negative chronotropic effect in the heart of the cod was equal to 700 ms, that of the frog--to 2.700 ms. Functional role of these differences is discussed in relation to the problem of the development of parasympathetic regulation of the heart rate in phylogenesis of vertebrates.

  9. Relationship between estradiol-17 beta seasonal profile and annual vitellogenin content of liver, fat body, plasma, and ovary in the frog (Rana esculenta).

    PubMed

    Varriale, B; Pierantoni, R; Di Matteo, L; Minucci, S; Milone, M; Chieffi, G

    1988-03-01

    The seasonal plasma estradiol-17 beta (E2-17 beta) profile and annual vitellogenin content of liver, fat body, plasma, and ovary were investigated in Rana esculenta. Concomitant with the increase in E2-17 beta, vitellogenin peaked in liver, plasma, and ovary during autumn and winter, while it remained at a relatively high concentration in fat body during spring. In vitro experiments showed that E2-17 beta (10(-9) M) is ineffective in inducing vitellogenin production in fat body, but is effective in inducing vitellogenin production in liver. As fat bodies do not produce the vitellogenin they contain, we suggest that fat bodies are involved in the transfer of vitellogenin to the ovary.

  10. A de novo Assembly of the Common Frog (Rana temporaria) Transcriptome and Comparison of Transcription Following Exposure to Ranavirus and Batrachochytrium dendrobatidis

    PubMed Central

    Price, Stephen J.; Garner, Trenton W. J.; Balloux, Francois; Ruis, Chris; Paszkiewicz, Konrad H.; Moore, Karen; Griffiths, Amber G. F.

    2015-01-01

    Amphibians are experiencing global declines and extinctions, with infectious diseases representing a major factor. In this study we examined the transcriptional response of metamorphic hosts (common frog, Rana temporaria) to the two most important amphibian pathogens: Batrachochytrium dendrobatidis (Bd) and Ranavirus. We found strong up-regulation of a gene involved in the adaptive immune response (AP4S1) at four days post-exposure to both pathogens. We detected a significant transcriptional response to Bd, covering the immune response (innate and adaptive immunity, complement activation, and general inflammatory responses), but relatively little transcriptional response to Ranavirus. This may reflect the higher mortality rates found in wild common frogs infected with Ranavirus as opposed to Bd. These data provide a valuable genomic resource for the amphibians, contribute insight into gene expression changes after pathogen exposure, and suggest potential candidate genes for future host-pathogen research. PMID:26111016

  11. Bullfrog tadpole (Rana catesbeiana) and red swamp crayfish (Procambarus clarkii) predation on early life stages of endangered razorback sucker (Xyrauchen texanus)

    USGS Publications Warehouse

    Mueller, G.A.; Carpenter, J.; Thornbrugh, D.

    2006-01-01

    Bullfrog tadpoles (Rana catesbeiana) and red swamp crayfish (Procambarus clarkii) are widespread introduced taxa that are problematic throughout the western United States. Their impact on native amphibians and crustaceans is well documented, but less is known regarding their influence on native fishes. Predator-prey tank tests showed both species consumed eggs and larvae of the endangered razorback sucker (Xyrauchen texanus) in a laboratory setting. Tadpoles consumed 2.2 razorback sucker eggs/d and 1.4 razorback sucker larvae/d, while crayfish ate 6.0 eggs/d and 3.5 larvae/d. Relatively high densities of bullfrog tadpoles and crayfish in razorback sucker spawning areas suggest that these nonnative taxa might pose a threat to the recruitment success of this and other imperiled native fish.

  12. Calcium oxalate nephrolithiasis and tubular necrosis in recent metamorphs of Rana sylvatica (Lithobates sylvaticus) fed spinach during the premetamorphic (tadpole) stage.

    PubMed

    Forzán, M J; Ferguson, L V; Smith, T G

    2015-03-01

    Amphibians in the family Ranidae (true frogs) seem highly susceptible to oxalosis, particularly when fed a diet high in oxalic acid during the premetamorphic (tadpole) stage. The authors describe the mortality of 150 captive-raised wood frogs (Rana sylvatica or Lithobates sylvaticus) from oxalate nephrolithiasis and renal tubular necrosis caused by consumption of boiled spinach during tadpole development. Renal lesions were due to intraluminal transparent crystals which were birefringent under polarized light and were identified morphologically and histochemically as composed of calcium oxalate. Evidence of early fibrosis or squamous metaplasia, and a presentation at least 2 weeks after spinach consumption had ended, suggested a subacute course. Tadpole-feeding protocols should avoid plants with high oxalate content (eg, spinach and rhubarb leaves), and any episode of high mortality in captive amphibians along with nephrolithiasis should prompt an evaluation of the feed sources for material with high oxalate content.

  13. Incidence and impact of axial malformations in larval bullfrogs (Rana catesbeiana) developing in sites polluted by a coal-burning power plant

    SciTech Connect

    Hopkins, W.A.; Congdon, J.; Ray, J.K.

    2000-04-01

    Amphibian malformations have recently received much attention from the scientific community, but few studies have provided evidence linking environmental pollution to larval amphibian malformations in the field. The authors document an increased incidence of axial malformations in bullfrog larvae (Rana catesbeiana) inhabiting two sites contaminated with coal combustion wastes. In the polluted sites, 18 and 37% of larvae exhibited lateral curvatures of the spine, whereas zero and 4% of larvae from two reference sites had similar malformations. Larvae from the most heavily polluted site had significantly higher tissue concentrations of potentially toxic trace elements, including As, Cd, Se, Cu, Cr, and V, compared with conspecifics from the reference sites. In addition, malformed larvae from the cost contaminated site had decreased swimming speeds compared with those of normal larvae from the same site. The authors hypothesize that the complex mixture of contaminants produced by coal combustion is responsible for the high incidence of malformations and associated effects on swimming performance.

  14. Diverse families of antimicrobial peptides isolated from skin secretions of three species of East Asian frogs, Babina daunchina, Babina adenopleura, and Rana omeimontis (Ranidae).

    PubMed

    Hu, Yuhong; Xu, Shiqi; Hu, Yonghong; Guo, Chao; Meng, Hao; Li, Jing; Liu, Jingze; Wang, Hui

    2014-07-01

    Twenty-two novel cDNAs encoding 22 peptide precursors for 19 mature peptides including antimicrobial peptides (AMPs) were identified from East Asian frog species Babina daunchina, Babina adenopleura, and Rana omeimontis skin-derived cDNA libraries. Two atypical members of the brevinin-1 family AMPs, named brevinin-1AN1 (FLTGVLKLASKIPSVLCAVLKTC) and brevinin-1DN1(FLKGVINLASKIPSMLCAVLKTC), were purified from the skin secretions of B. adenopleura and B. daunchina, respectively. A member of the ranatuerin-2 family AMP named ranatuerin-2DN1 (GLFDSITQGLKDTAVKLLDKIKCKLSACPPA) was also purified from the skin secretion of B. daunchina. One AMP named japonicin-2OM1 (FIVPSIFLLKKAFCIALKKNC) was purified from the skin secretion of R. omeimontis. The antimicrobial tests showed that brevinin-1DN1, brevinin-1DN2, brevinin-1AN1, and japonicin-2OM1 possess higher antimicrobial activity against Gram-positive bacteria than Gram-negative bacteria.

  15. [Analysis of helminthofauna of common spaedfoot Pelobates fuscus (Laurenti, 1768) and moor frog Rana arvalis Nilsson, 1842 (Amphibia: Anura) at their joint habitation].

    PubMed

    Ruchin, A B; Chikhliaev, I V; Lukiianov, S V

    2009-01-01

    The helminths fauna of common spaedfoot Pelobates fuscus (Laurenti, 1768) and moor frog Rana arvalis Nilsson, 1842 has been studied at their joint habitation. The stuff was collected in 1998-2002, 2004-2006 years in several regions (republic Mordovia, Samara and Saratov areas). The processing of a stuff is conducted by a method of full helmintologic dissecting. The fauna of helminths considerably differs. For common spaedfoot only 13 species of helminths was detected which also parasitized moor frog (for moor frog 23 species) are detected. The index Jaccar demonstrated mean resemblance structure of helminths and varied from 0.25 till 0.69, and the index Morisite--from 44.58 of % till 74.51 of %. The communities of parasites of common spaedfoot was characterized by low values of an index of Shannon, but the high indexes of an index Simpson, whereas for moor frog tracked the return tendence.

  16. Immunofluorescence studies on gonadotropin releasing hormone (GRH) in the fore-brain and the neurohypophysis of the green frog, Rana esculenta L.

    PubMed

    Goos, H J; Ligtenberg, P J; van Oordt, P G

    1976-05-13

    Using antibodies against mammalian LH-RH, the double antibody-immunofluorecence technique has been applied to serial cross sections of the brains of adult Rana esculenta. Immunoreactive material was found in perikarya of an unpaired nucleus in front of the preoptic recess. The axons of these perikarya also contain fluorescing material. They form a single bundle which passes under the preoptic recess, than splits into two tracts, one on either side of the optic chiasm. The two tracts reunite just before entering the median eminence. The axons end near the capillaries in the outer zone of the median eminence. The possibility of two separate centres for the stimulation of gonadotropic activity in the brains of anurans is discussed.

  17. Sequencing and analysis of the internal transcribed spacers (ITSs) and coding regions in the EcoR I fragment of the ribosomal DNA of the Japanese pond frog Rana nigromaculata.

    PubMed

    Sumida, Masayuki; Kato, Yoji; Kurabayashi, Atsushi

    2004-04-01

    The rDNA of eukaryotic organisms is transcribed as the 40S-45S rRNA precursor, and this precursor contains the following segments: 5' - ETS - 18S rRNA - ITS 1 - 5.8S rRNA - ITS 2 - 28S rRNA - 3'. In amphibians, the nucleotide sequences of the rRNA precursor have been completely determined in only two species of Xenopus. In the other amphibian species investigated so far, only the short nucleotide sequences of some rDNA fragments have been reported. We obtained a genomic clone containing the rDNA precursor from the Japanese pond frog Rana nigromaculata and analyzed its nucleotide sequence. The cloned genomic fragment was 4,806 bp long and included the 3'-terminus of 18S rRNA, ITS 1, 5.8S rRNA, ITS 2, and a long portion of 28S rRNA. A comparison of nucleotide sequences among Rana, the two species of Xenopus, and human revealed the following: (1) The 3'-terminus of 18S rRNA and the complete 5.8S rRNA were highly conserved among these four taxa. (2) The regions corresponding to the stem and loop of the secondary structure in 28S rRNA were conserved between Xenopus and Rana, but the rate of substitutions in the loop was higher than that in the stem. Many of the human loop regions had large insertions not seen in amphibians. (3) Two ITS regions had highly diverged sequences that made it difficult to compare the sequences not only between human and frogs, but also between Xenopus and Rana. (4) The short tracts in the ITS regions were strictly conserved between the two Xenopus species, and there was a corresponding sequence for Rana. Our data on the nucleotide sequence of the rRNA precursor from the Japanese pond frog Rana nigromaculata were used to examine the potential usefulness of the rRNA genes and ITS regions for evolutionary studies on frogs, because the rRNA precursor contains both highly conserved regions and rapidly evolving regions.

  18. Long-term observation of amphibian populations inhabiting urban and forested areas in Yekaterinburg, Russia

    NASA Astrophysics Data System (ADS)

    Vershinin, Vladimir L.; Vershinina, Svetlana D.; Berzin, Dmitry L.; Zmeeva, Darya V.; Kinev, Alexander V.

    2015-05-01

    This article presents data derived from a 36 year-long uninterrupted observational study of amphibian populations living in the city and vicinity of Yekaterinburg, Russia. This area is inhabited by six amphibian species. Based on a degree of anthropogenic transformation, the urban territory is divided into five highly mosaic zones characterized by vegetation, temperature, and a distinctive water pollution profile. Population data is presented year-by-year for the number of animals, sex ratio, and species-specific fecundity including the number and quality of spawns for the following amphibian species: Salamandrella keyserligii, Rana arvalis, R. temporaria, Lissotriton vulgaris, and Pelophylax ridibundus. These data provide an excellent opportunity to assess an urban environment from an animal population-wide perspective, as well as revealing the forces driving animal adaptation to the anthropogenic transformation of habitats.

  19. Long-term observation of amphibian populations inhabiting urban and forested areas in Yekaterinburg, Russia

    PubMed Central

    Vershinin, Vladimir L.; Vershinina, Svetlana D.; Berzin, Dmitry L.; Zmeeva, Darya V.; Kinev, Alexander V.

    2015-01-01

    This article presents data derived from a 36 year-long uninterrupted observational study of amphibian populations living in the city and vicinity of Yekaterinburg, Russia. This area is inhabited by six amphibian species. Based on a degree of anthropogenic transformation, the urban territory is divided into five highly mosaic zones characterized by vegetation, temperature, and a distinctive water pollution profile. Population data is presented year-by-year for the number of animals, sex ratio, and species-specific fecundity including the number and quality of spawns for the following amphibian species: Salamandrella keyserligii, Rana arvalis, R. temporaria, Lissotriton vulgaris, and Pelophylax ridibundus. These data provide an excellent opportunity to assess an urban environment from an animal population-wide perspective, as well as revealing the forces driving animal adaptation to the anthropogenic transformation of habitats. PMID:25984350

  20. Decreased winter severity increases viability of a montane frog population

    PubMed Central

    McCaffery, Rebecca M.; Maxell, Bryce A.

    2010-01-01

    Many proximate causes of global amphibian declines have been well documented, but the role that climate change has played and will play in this crisis remains ambiguous for many species. Breeding phenology and disease outbreaks have been associated with warming temperatures, but, to date, few studies have evaluated effects of climate change on individual vital rates and subsequent population dynamics of amphibians. We evaluated relationships among local climate variables, annual survival and fecundity, and population growth rates from a 9-year demographic study of Columbia spotted frogs (Rana luteiventris) in the Bitterroot Mountains of Montana. We documented an increase in survival and breeding probability as severity of winter decreased. Therefore, a warming climate with less severe winters is likely to promote population viability in this montane frog population. More generally, amphibians and other ectotherms inhabiting alpine or boreal habitats at or near their thermal ecological limits may benefit from the milder winters provided by a warming climate as long as suitable habitats remain intact. A more thorough understanding of how climate change is expected to benefit or harm amphibian populations at different latitudes and elevations is essential for determining the best strategies to conserve viable populations and allow for gene flow and shifts in geographic range. PMID:20421473

  1. A generic weather-driven model to predict mosquito population dynamics applied to species of Anopheles, Culex and Aedes genera of southern France.

    PubMed

    Ezanno, P; Aubry-Kientz, M; Arnoux, S; Cailly, P; L'Ambert, G; Toty, C; Balenghien, T; Tran, A

    2015-06-01

    An accurate understanding and prediction of mosquito population dynamics are needed to identify areas where there is a high risk of mosquito-borne disease spread and persistence. Simulation tools are relevant for supporting decision-makers in the surveillance of vector populations, as models of vector population dynamics provide predictions of the greatest risk periods for vector abundance, which can be particularly helpful in areas with a highly variable environment. We present a generic weather-driven model of mosquito population dynamics, which was applied to one species of each of the genera Anopheles, Culex, and Aedes, located in the same area and thus affected by similar weather conditions. The predicted population dynamics of Anopheles hyrcanus, Culex pipiens, and Aedes caspius were not similar. An. hyrcanus was abundant in late summer. Cx. pipiens was less abundant but throughout the summer. The abundance of both species showed a single large peak with few variations between years. The population dynamics of Ae. caspius showed large intra- and inter-annual variations due to pulsed egg hatching. Predictions of the model were compared to longitudinal data on host-seeking adult females. Data were previously obtained using CDC-light traps baited with carbon dioxide dry ice in 2005 at two sites (Marais du Viguerat and Tour Carbonnière) in a favourable temperate wetland of southern France (Camargue). The observed and predicted periods of maximal abundance for An. hyrcanus and Cx. pipiens tallied very well. Pearson's coefficients for these two species were over 75% for both species. The model also reproduced the major trends in the intra-annual fluctuations of Ae. caspius population dynamics, with peaks occurring in early summer and following the autumn rainfall events. Few individuals of this species were trapped so the comparison of predicted and observed dynamics was not relevant. A global sensitivity analysis of the species-specific models enabled us to

  2. Patch Clamp on the Luminal Membrane of Exocrine Gland Acini from Frog Skin (Rana esculenta) Reveals the Presence of Cystic Fibrosis Transmembrane Conductance Regulator–like Cl− Channels Activated by Cyclic AMP

    PubMed Central

    Sørensen, Jakob Balslev; Larsen, Erik Hviid

    1998-01-01

    Chloride channels in the luminal membrane of exocrine gland acini from frog skin (Rana esculenta) constituted a single homogeneous population. In cell-attached patches, channels activated upon exposure to isoproterenol, forskolin, or dibutyryl-cAMP and isobutyl-1-methyl-xanthine rectified in the outward direction with a conductance of 10.0 ± 0.4 pS for outgoing currents. Channels in stimulated cells reversed at 0 mV applied potential, whereas channels in unstimulated cells reversed at depolarized potentials (28.1 ± 6.7 mV), indicating that Cl− was above electrochemical equilibrium in unstimulated, but not in stimulated, cells. In excised inside-out patches with 25 mM Cl− on the inside, activity of small (8-pS) linear Cl−-selective channels was dependent upon bath ATP (1.5 mM) and increased upon exposure to cAMP-dependent protein kinase. The channels displayed a single substate, located just below 2/3 of the full channel amplitude. Halide selectivity was identified as PBr > PI > PCl from the Goldman equation; however, the conductance sequence when either halide was permeating the channel was GCl > GBr >> GI. In inside-out patches, the channels were blocked reversibly by 5-nitro-2-(3-phenylpropylamino)benzoic acid, glibenclamide, and diphenylamine-2-carboxylic acid, whereas 4,4-diisothiocyanatostilbene-2,2-disulfonic acid blocked channel activity completely and irreversibly. Single-channel kinetics revealed one open state (mean lifetime = 158 ± 72 ms) and two closed states (lifetimes: 12 ± 4 and 224 ± 31 ms, respectively). Power density spectra had a double-Lorentzian form with corner frequencies 0.85 ± 0.11 and 27.9 ± 2.9 Hz, respectively. These channels are considered homologous to the cystic fibrosis transmembrane conductance regulator Cl− channel, which has been localized to the submucosal skin glands in Xenopus by immunohistochemistry (Engelhardt, J.F., S.S. Smith, E. Allen, J.R. Yankaskas, D.C. Dawson, and J.M. Wilson. 1994. Am. J. Physiol. 267

  3. Inbreeding and road effect zone in a Ranidae: the case of Agile frog, Rana dalmatina Bonaparte, 1840.

    PubMed

    Lesbarrères, David; Pagano, Alain; Lodé, Thierry

    2003-08-01

    Inbreeding has often been invoked in the extinction of local populations. In eleven western France populations of Agile frog studied, observed heterozygosity was significantly lower than expected in all cases, giving new evidence of such a depression in small populations. It especially occurred in ponds located near an highway rather than in undisturbed populations (FIS = 0.544 and 0.315, respectively). Thus, our results argue for a "road effect zone". Discussing about road distance and conservation policies, we propose that roads are directly involved in inbreeding and in local extinction. Thus, road construction ought to consider conservation management.

  4. Breeding site heterogeneity reduces variability in frog recruitment and population dynamics

    USGS Publications Warehouse

    McCaffery, Rebecca M.; Eby, Lisa A.; Maxell, Bryce A.; Corn, Paul Stephen

    2013-01-01

    Environmental stochasticity can have profound effects on the dynamics and viability of wild populations, and habitat heterogeneity provides one mechanism by which populations may be buffered against the negative effects of environmental fluctuations. Heterogeneity in breeding pond hydroperiod across the landscape may allow amphibian populations to persist despite variable interannual precipitation. We examined recruitment dynamics over 10 yr in a high-elevation Columbia spotted frog (Rana luteiventris) population that breeds in ponds with a variety of hydroperiods. We combined these data with matrix population models to quantify the consequences of heterogeneity in pond hydroperiod on net recruitment (i.e. number of metamorphs produced) and population growth rates. We compared our heterogeneous system to hypothetical homogeneous environments with only ephemeral ponds, only semi-permanent ponds, and only permanent ponds. We also examined the effects of breeding pond habitat loss on population growth rates. Most eggs were laid in permanent ponds each year, but survival to metamorphosis was highest in the semi-permanent ponds. Recruitment success varied by both year and pond type. Net recruitment and stochastic population growth rate were highest under a scenario with homogeneous semi-permanent ponds, but variability in recruitment was lowest in the scenario with the observed heterogeneity in hydroperiods. Loss of pond habitat decreased population growth rate, with greater decreases associated with loss of permanent and semi-permanent habitat. The presence of a diversity of pond hydroperiods on the landscape will influence population dynamics, including reducing variability in recruitment in an uncertain climatic future.

  5. Molecular detection of Wolbachia pipientis in natural populations of mosquito vectors of Dirofilaria immitis from continental Portugal: first detection in Culex theileri.

    PubMed

    DE Pinho Mixão, V; Mendes, A M; Maurício, I L; Calado, M M; Novo, M T; Belo, S; Almeida, A P G

    2016-09-01

    Wolbachia pipientis (Rickettsiales: Rickettsiaceae) protects mosquitoes from infections with arboviruses and parasites. However, the effect o