Occurrence of diploid ova in Rana pipiens
DOE Office of Scientific and Technical Information (OSTI.GOV)
Richards, C.M.; Nace, G.W.
All 12 Rana pipiens females tested from three populations produced diploid ova. These were identified by the development of from 0.15 to 35 percent per clutch of normal diploid-like embryos among large numbers of haploid embryos following activation of R. pipiens eggs with irradiated R. clamitans sperm. Their diploid nuclear constitution was demonstrated by diploid cell size at Shumway Stage 19, and by the diploid number of chromosomes both as embryos and as mature frogs, and was confirmed by the occurrence of triploid embryos among normally fertilized progeny from the same female parents. Although the precise cytogenetic events leading tomore » the origin of these diploid ova were not directly determined, we were led to conclude that, although diploid ova may result from polynucleate oocytes, the diploidy reported here was an expression of abnormal meiosis that occurred under genetic control. Such anomalies, at the orders of frequency we observed in R. pipiens, have important consequences for all studies using Anuran model systems.« less
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.
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.
A survey of blood and other tissue parasites of leopard frogs Rana pipiens in the United States.
Levine, N D; Nye, R R
1977-01-01
In a survey of blood and other tissue parasites from 137 leopard frogs, Rana pipiens complex, purchased from 13 commercial vendors in 8 states in the United States, Trypanosoma pipientis was found in 2 R. p. berlandieri, Toxoplasma ranae in 1 R. pipiens, Isospora lieberkuehni in 1 leopard frog, Haemogregarina magna in 44, Lankesterella minima in 3, Leptotheca ohlmacheri in 3 and microfilariae of Foleyella sp. in 6. The report of I. lieberkuehni is presumably a new host record. Haemogregarina temporariae (Nöller,, 1920) nov. comb. is established as a new combination for Nematopsis temporariae.
Laboratory studies bearing on pigment pattern polymorphisms in wild populations of Rana pipiens.
Merrell, D J
1972-01-01
Data are presented for 2,393 progeny from a number of crosses related to a study in ecological genetics of the Burnsi and Kandiyohi polymorphisms in natural populations of the leopard frog, Rana pipiens. No significant differences in viability were found between wild-type homozygotes (+/+) and Burnsi heterozygotes (B/+) or homozygotes (B/B). Similarly, no difference in viability was found between wild-type (+/+) and Kandiyohi heterozygotes (K/+) and homozygotes (K/K). However, there appears to be slight reduction in viability of the double dominant heterozygote (B/+; K/+) in comparison with (+/+), (B/+), and (K/+) progeny from the same cross.-The Kandiyohi heterozygotes (K/+) appeared to have a more rapid rate of development from fertilization through metamorphosis than wild-type (+/+) or Burnsi (B+/) or Burnsi-Kandiyohi heterozygotes (B+K/+). Since Kandiyohi is associated primarily with the prairie habitat (Merrell 1965), this finding suggests that the adaptive advantage of Kandiyohi lies in the more rapid rate of development of frogs carrying this gene, enabling them to complete metamorphosis before the prairie breeding ponds dry up.-Data are presented from crosses involving dorsal spot number. The results suggest that heredity plays a role in the determination of dorsal spot number but that non-genetic influences are also of considerable importance.-The results of these crosses are discussed with respect to their bearing on the formation of pigment patterns in Rana pipiens. From the available data it is clear that the pigment pattern in Rana pipiens is a complex trait influenced by major gene loci, by modifying genes, and by environmental effects. The relative importance of these factors varies depending on the particular combination of genetic and environmental conditions.
Hogan, Natacha S; Lean, David R S; Trudeau, Vance L
2006-08-01
Endocrine disrupting compounds (EDCs) are often detected in the aquatic environment and can negatively affect the health of wildlife populations. However, little is known about the sensitivity of native amphibians to EDCs. Wood frogs (Rana sylvatica) and Northern leopard frogs (Rana pipiens) were exposed to three estrogenic EDCs: estradiol (E2), ethinylestradiol (EE2), and 4-tert-octylphenol (OP). In addition, R. pipiens were exposed during two developmental stages (Gosner stages 26 and 36) to examine life-stage differences in sensitivity. Tadpoles were exposed for 2 wk to 8 nominal concentrations (0.25 microM-10 microM) of each compound. Individual mortality was recorded during the exposure period, while body weight was measured at the end of 2 wk. LC50 values were calculated, and differences in body weight between vehicle control and exposed groups were assessed. Rank order toxicity of the compounds for both R. pipiens stages and both species was OP > EE2 > E2. Gosner stage 26 tadpoles were more sensitive (LC50: E2 [5.57 microM], EE2 [3.01 microM], OP [1.36 microM]) to all three compounds when compared to stage 36 tadpoles (LC50: E2 [>10 microM], EE2 [4.17 microM], OP [2.80 microM]). Interspecies comparisons revealed R. sylvatica tadpoles (LC50: E2 [2.50 microM], EE2 [1.89 microM], OP [0.74 microM]) as being more sensitive to the three compounds than R. pipiens (LC50: E2 [4.56 microM], EE2 [2.75 microM], OP [1.42 microM]). Xenoestrogen exposure also affected tadpole body weight which may have long-term adverse effects on the rate of metamorphosis. These results provide toxicological data needed for assessing sublethal effects of estrogenic compounds on amphibian development and suggest that environmental levels of OP may pose a serious risk to the health of amphibian populations.
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...
Leney, Jocelyn L; Drouillard, Ken G; Haffner, G Douglas
2006-11-01
Polychlorinated biphenyl (PCB) elimination rates were used to determine whether green frogs (Rana clamitans) and leopard frogs (Rana pipiens) at three life stages (tadpole, metamorph, and adult) were capable of metabolic biotransformation of PCBs. Polychlorinated biphenyls were grouped according to their structural properties to determine whether congeners with specific structures were eliminated more rapidly than others, which would suggest the occurrence of metabolic biotransformation. For both species and at all life stages studied, PCBs with adjacent unsubstituted meta-para sites (group-2 PCBs) were eliminated significantly faster than PCBs with adjacent unsubstituted ortho-meta sites (group-3 PCBs). Because these groups of PCBs span similar hydrophobicity ranges, it was concluded that green and leopard frogs can selectively metabolize group-2 PCBs. Metabolic activity was more pronounced in the metamorph stage than in any other life stage. In green frog metamorphs, the average elimination rate of the group-2 PCBs was 4.9-fold greater than that of the group-3 PCBs. By contrast, in green frog tadpoles, group-2 PCBs were eliminated 1.9-fold faster than group-3 PCBs, and in green frog adults, group-2 PCBs were eliminated 1.5-fold faster than group-3 PDBs.
Photoinduced toxicity of fluoranthene to northern leopard frogs (Rana pipiens)
DOE Office of Scientific and Technical Information (OSTI.GOV)
Monson, P.D.; Call, D.J.; Cox, D.A.
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 {+-} 0.05more » {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.« less
DOE Office of Scientific and Technical Information (OSTI.GOV)
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 126more » 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.« less
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.
Ankley, Gerald T; Kuehl, Douglas W; Kahl, Michael D; Jensen, Kathleen M; Butterworth, Brian C; Nichols, John W
2004-11-01
A number of recent monitoring studies have demonstrated elevated concentrations of perfluorooctanesulfonate (PFOS) in humans and wildlife throughout the world. Although no longer manufactured in the United States, the global distribution and relative persistence of PFOS indicates a need to understand its potential ecological effects. Presently, little is known concerning toxicity of PFOS in chronic exposures with aquatic species. Therefore, we evaluated the effects of PFOS on survival and development of the northern leopard frog (Rana pipiens) from early embryogenesis through complete metamorphosis. Exposures were conducted via water at measured PFOS concentrations ranging from 0.03 to 10 mg/L. Animals exposed to 10 mg/L began dying within approximately two weeks of test initiation. Survival was not affected by PFOS at lower concentrations; however, time to metamorphosis was delayed and growth reduced in the 3-mg/L treatment group. Tadpoles readily accumulated PFOS directly from water. Using a one-compartment bioaccumulation model, growth was shown to have a modest impact on steady-state PFOS concentrations. Variability in observed growth rates and the possible contribution of a size-dependent decrease in PFOS elimination rate contributed uncertainty to modeling efforts. Nevertheless, fitted uptake and elimination rate constants were comparable to those determined in earlier studies with juvenile rainbow trout. Overall, our studies suggest that R. pipiens is not exceptionally sensitive to PFOS in terms of either direct toxicity or bioconcentration potential of the chemical.
DOE Office of Scientific and Technical Information (OSTI.GOV)
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 themore » 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.« less
Stanley, Jacob K; Lotufo, Guilherme R; Biedenbach, James M; Chappell, Pornsawan; Gust, Kurt A
2015-04-01
An initiative within the US military is targeting the replacement of traditional munitions constituents with insensitive munitions to reduce risk of accidental detonation. The purpose of the present study was to comparatively assess toxicity of the traditional munitions constituents 2,4,6-trinitrotoluene (TNT) and 1,3,5-trinitroperhydro-1,3,5-triazine (RDX) with the new insensitive munitions constituents 2,4-dinitroanisole (DNAN) and 3-nitro-1,2,4-triazol-5-one (NTO). The following exposure durations were performed with Rana pipiens (leopard frog) tadpoles: TNT and DNAN, 96 h and 28 d; RDX, 10 d and 28 d; NTO, 28 d. The 96-h 50% lethal concentration (LC50) values and 95% confidence intervals for TNT and DNAN were 4.4 mg/L (4.2 mg/L, 4. 7 mg/L) and 24.3 mg/L (21.3 mg/L, 27.6 mg/L), respectively. No significant impacts on survival were observed in the 10-d exposure to RDX up to 25.3 mg/L. Effects on tadpole swimming distance were observed with a lowest-observed-effect concentration (LOEC) of 5.9 mg/L RDX. In the 28-d exposures, the LOECs for survival for TNT, DNAN, and NTO were 0.003 mg/L, 2.4 mg/L, and 5.0 mg/L, respectively. No significant mortality was observed in the RDX chronic 28-d exposure up to the highest treatment level tested of 28.0 mg/L. Neither tadpole developmental stage nor growth was significantly affected in any of the 28-d exposures. Rana pipiens were very sensitive to chronic TNT exposure, with an LOEC 3 orders of magnitude lower than those for insensitive munitions constituents DNAN and NTO. © 2015 SETAC.
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 3133) 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 (47.597.5%), 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.
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.
Hernández, C J; Ortíz, T; Rosa, C; Foster, K; Tyagi, M; Lugo, N; Albrecht, R; Chinapen, S
2007-04-01
Mucociliary activity is an important clearance mechanism in the respiratory system of air breathing vertebrates. Substance P (SP) and acetylcholine play a key role in the stimulation of the mucociliary transport in the frog palate. In this study, retrograde neuronal tracing was combined with immunocytochemistry for SP and choline acetyl transferase (ChAT) in the trigeminal ganglion and for neurokinin-1 receptor (NK1R) in the palate of Rana pipiens. The cells of origin of the palatine nerve were identified in the trigeminal ganglion using the retrograde tracer Fluorogold (FG). Optimal labeling of FG cells in the trigeminal ganglion was obtained at 96 h of exposure. Immunoflorescent shows that SP and acetylcholine are co-localized in 92% of the cells labeled with FG in the trigeminal ganglion. NK1 receptors were found in the membrane of epithelial and goblet cells of the palate. Ultrastructural study of the palate showed axonal-like endings with vesicles in connection with epithelial and goblet cells. These results further support the concerted action of both neurotransmitters in the regulation of mucociliary activity in the frog palate.
Clark, H. Fred; Brennan, James C.; Zeigel, Robert F.; Karzon, David T.
1968-01-01
Viruses were isolated from kidneys of normal and renal tumor-bearing Vermont Rana pipiens after subinoculation into red eft newts (Triturus viridescens). Organs of efts inoculated with viable cell suspensions from four of seven tumor-bearing kidneys yielded virus (LT-1, -2, -3, -4) when inoculated into TH-1 (Terrapene heart) cell culture. One tumor-bearing kidney also yielded virus (L-4) by direct inoculation into TH-1 cells. An additional isolate (L-5) was obtained from 1 of 52 normal Vermont frog kidneys inoculated directly into TH-1 cells. LT-1 was propagated with cytopathic effect (CPE) in each of 38 cell types tested, of fish, amphibian, reptilian, avian, and mammalian origin, at 23 or 30 C. LT-1 through LT-4, L-4 and L-5, and FV-1 through FV-3 each induced similar CPE in all cells tested. LT-2, however, induced CPE that progressed at a slower rate than that caused by the other isolates and produced smaller plaques (<0.8 mm) under starch gel overlay. Each of the viruses replicated to high titer in embryonated eggs incubated at 30 C. The viruses also grew in efts and adult newts, but not in bullfrog (Rana catesbeiana) tadpoles or adult leopard frogs. Tumor induction in adult leopard frogs inoculated with LT-1 was not demonstrated. Electron microscopic observations of LT-1 and LT-2 viruses revealed cytoplasmic particles, hexagonal in cross section, approximately 120 to 140 mμ in diameter, containing a dense nucleoid. LT-1 and LT-2 viruses were indistinguishable from FV-1 and Tipula iridescent virus. LT-1 was presumed to be a deoxyribonucleic acid virus on the basis of 5-bromodeoxyuridine inhibition. The isolates were ether-sensitive. On the basis of biological, physicochemical, and antigenic similarities, LT-1 through LT-4, L-4, L-5, FV-1 through FV-3, and isolates recently recovered from the bullfrog and the newt may represent strains of the same amphibian cytoplasmic virus. Images PMID:4972302
Effects of atrazine on embryos, larvae, and adults of anuran amphibians.
Allran, J W; Karasov, W H
2001-04-01
We examined the effects of atrazine (0-20 mg/L) on embryos, larvae, and adult anuran amphibian species in the laboratory. Atrazine treatments did not affect hatchability of embryos or 96-h posthatch mortality of larvae of Rana pipiens, Rana sylvatica, or Bufo americanus. Furthermore, atrazine had no effect on swimming speed (measured for R. pipiens only). However, there was a dose-dependent increase in deformed larvae of all three species with increasing atrazine concentration. In adult R. pipiens, atrazine increased buccal and thoracic ventilation, indicating respiratory distress. However, because atrazine had no affect on hemoglobin, this respiratory distress was probably not indicative of reduced oxygen-carrying capacity of the blood. Frogs exposed to the highest atrazine concentration stopped eating immediately after treatment began and did not eat during the 14-d experiment. However, no decreases in mass were measured even for frogs that were not eating, probably because of compensatory fluid gain from edema. Atrazine concentrations found to be deleterious to amphibian embryos and adults are considerably higher than concentrations currently found in surface waters in North America. Therefore, direct toxicity of atrazine is probably not a significant factor in recent amphibian declines.
Oxidative stress induced in PCB 126-exposed northern leopard frogs, Rana pipiens
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
Oxidative stress induced in PCB 126-exposed northern leopard frogs, Rana pipiens.
Huang, Yue-wern; Hoffman, David J; Karasov, William H
2007-04-15
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.
Hernández, C. J.; Ortíz, T.; Foster, C. Rosa K.; Tyagi, M; Lugo, N.; Albrecht, R.; Chinapen, S.
2007-01-01
Mucociliary activity is an important clearance mechanism in the respiratory system of air breathing vertebrates. Substance P (SP) and acetylcholine play a key role in the stimulation of the mucociliary transport in the frog palate. In this study, retrograde neuronal tracing was combined with immunocytochemistry for SP and choline acetyl transferase (ChAT) in the trigeminal ganglion and for neurokinin-1 receptor (NK1R) in the palate of Rana pipiens. The cells of origin of the palatine nerve were identified in the trigeminal ganglion using the retrograde tracer Fluorogold (FG). Optimal labeling of FG cells in the trigeminal ganglion was obtained at 96 h of exposure. Immunoflorescent shows that SP and acetylcholine are co-localized in 92% of the cells labeled with FG in the trigeminal ganglion. NK1 receptors were found in the membrane of epithelial and goblet cells of the palate. Ultrastructural study of the palate showed axonal-like endings with vesicles in connection with epithelial and goblet cells. These results further support the concerted action of both neurotransmitters in the regulation of mucociliary activity in the frog palate. PMID:17276713
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
AMBIENT SOLAR UV RADIATION CAUSES MORTALITY IN LARVAE OF THREE SPECIES OF RANA
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...
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.
Crump, Douglas; Lean, David; Trudeau, Vance L
2002-03-01
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
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
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.
Langlois, Valérie S; Carew, Amanda C; Pauli, Bruce D; Wade, Michael G; Cooke, Gerard M; Trudeau, Vance L
2010-04-01
There are conflicting reports regarding the effects of atrazine (ATZ) on amphibian development. Therefore, further studies are needed to examine the potential mechanisms of action of ATZ in amphibians. Our aim in this study was to determine whether low concentrations of ATZ affect gonadal development and metamorphosis in the Northern leopard frog, Rana pipiens. Tadpoles were exposed in outdoor mesocosms to nominal concentrations of 0.1 and 1.8 microg/L of formulated ATZ from Gosner stage 27 (G27) to metamorphic climax (G42). Exposure to 17alpha-ethinylestradiol (EE2; 1.5 microg/L) provided a positive control for induction of testicular oocytes in males. Endocrine-related gene expression and gonadal histopathology were examined at G42 and in a subset of premetamorphic G34 tadpoles that failed to metamorphose. Gonadal gross morphology revealed that the 1.8-microg/L ATZ treatment produced 20% more females compared with the control. Histologic analysis revealed that 22% of EE2-treated males had testicular oocytes, whereas none were observed in any animals from the control or either ATZ groups. ATZ increased brain estrogen receptor alpha mRNA to 2.5 times that of the control at premetamorphosis and altered liver levels of 5beta-reductase activity at metamorphosis. In contrast, brain aromatase mRNA level and activity did not change. ATZ treatments significantly reduced metamorphic success (number of animals reaching metamorphosis) without affecting body weight, snout-vent length, or age at metamorphosis. Gene expression analysis indicated that ATZ decreased the expression of deiodinase type 3 in the tail at premetamorphosis. Our study indicates that exposure to low concentrations of ATZ in experimental mesocosms alters gonadal differentiation and metamorphosis in developing R. pipiens.
Hogan, Natacha S; Duarte, Paula; Wade, Michael G; Lean, David R S; Trudeau, Vance L
2008-05-01
During the transformation from larval tadpole to juvenile frog, there are critical periods of metamorphic development and sex differentiation that may be particularly sensitive to endocrine disruption. The aim of the present study was to identify sensitive developmental periods for estrogenic endocrine disruption in the northern leopard frog (Rana pipiens) using short, targeted exposures to the synthetic estrogen, ethinylestradiol (EE2). Post-hatch tadpoles (Gosner stage 27) were exposed over five distinct periods of metamorphosis: early (stage 27-30), mid (stage 30-36), early and mid (stage 27-36), late (stage 36-42), and the entire metamorphic period (chronic; stage 27-42). For each period, animals were sampled immediately following the EE2 exposure and at metamorphic climax (stage 42). The effects of EE2 on metamorphic development and sex differentiation were assessed through measures of length, weight, developmental stage, days to metamorphosis, sex ratios and incidence of gonadal intersex. Our results show that tadpoles exposed to EE2 during mid-metamorphosis were developmentally delayed immediately following exposure and took 2 weeks longer to reach metamorphic climax. In the unexposed groups, there was low proportion (0.15) of intersex tadpoles at stage 30 and gonads appeared to be morphologically distinct (male and female) in all individuals by stage 36. Tadpoles exposed early in development displayed a strong female-biased sex ratio compared to the controls. Moreover, these effects were also seen at metamorphic climax, approximately 2-3 months after the exposure period, demonstrating that transient early life-stage exposure to estrogen can induce effects on the reproductive organs that persist into the beginning of adult life-stages.
Bioefficacy of Some Egyptian Aromatic Plants on Culex pipiens (Diptera: Culicidae) Adults and Larvae
El Zayyat, Elham A; Soliman, Mohammed I; Elleboudy, Noha A; Ofaa, Shaimaa E
2017-01-01
Background: Protecting the environment from chemical hazards of synthetic insecticides along with offering of new breeding areas for vectors by urbanization indicate the trial of natural insecticides. Methods: The acetone extracts of Anethum graveolens, Ocimum basilicum and Thymus vulgaris were tested for their insecticidal effect on Culex pipiens adults and larvae in different concentrations depending on the technique used. Results: The extracts were significantly effective in all models used with basil being the best in all tested three techniques (LC50= 0.064) in larval feeding, (LC50= 0.330) in CDC bottle assay and (LC50= 13.148) in adults feeding (P< 0.05). Conclusion: The results recommend the eco-friendly studied extracts as candidates for controlling Cx. pipiens the lymphatic filariasis vector. PMID:29026862
Parasites of the mink frog (rana septentrionalis) from Minnesota, U.S.A.
Schotthoefer, Anna M.; Bolek, M.G.; Cole, Rebecca A.; Beasley, Val 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.
Pseudacris triseriata (western chorus frog) and Rana sylvatica (wood frog) chytridiomycosis
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.
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.
DOE Office of Scientific and Technical Information (OSTI.GOV)
Huang, Y.; Jung, R.E.; Karasov, W.H.
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,more » 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.« less
Cakir, Yavuz; Strauch, Stephen M
2005-01-01
Tricaine (MS-222) is used commonly for sedation, immobilization, and anesthesia of poikilothermic animals. The anesthetic efficacy of different concentrations of MS-222 was compared to benzocaine and pentobarbital on the physiological changes, heart rate and ECG (electrocardiogram) parameters in the leopard frog, Rana pipiens. Loss of righting reflex (RR), loss of pain response (NR = nociceptor response) and recovery time were measured. Heart rate and ECG parameters were also tested before and during anesthesia. The time to loss of RR and NR decreased while recovery time markedly increased with the increasing concentration of MS-222. Benzocaine at 200 mg/l induced a rapid anesthesia, but all frogs needed resuscitation. Pentobarbital at 300 mg/l induced a slow anesthesia, however, all of the frogs also needed resuscitation. All anesthetics at the mentioned concentrations decreased heart rate significantly as well as altered the ECG parameters. All anesthetics prolonged the Q-T interval, and MS-222 at 800 mg/l and benzocaine at 200 mg/l were the most effective anesthetic concentrations in increasing the Q-T interval. Frogs anesthetized by benzocaine and pentobarbital and high concentrations of MS-222 required resuscitation due to hypoxia. Pentobarbital and benzocaine seem to be very effective compounds, but their safety margins are narrow because of ventilatory failure. Therefore, MS-222 at a concentration of 200 mg/l or less is highly recommended for leopard frogs because prolonged recovery, high mortality rate and significant ECG changes are observed with higher concentrations of MS-222.
Lotufo, Guilherme R; Biedenbach, James M; Sims, Jerre G; Chappell, Pornsawan; Stanley, Jacob K; Gust, Kurt A
2015-04-01
The manufacturing of explosives and their loading, assembling, and packing into munitions for use in testing on training sites or battlefields has resulted in contamination of terrestrial and aquatic sites that may pose risk to populations of sensitive species. The bioaccumulative potential of the conventional explosives 2,4,6-trinitrotoluene (TNT) and hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) and of the insensitive munitions (i.e., less shock sensitive) compound 2,4-dinitroanisole (DNAN) were assessed using the Northern leopard frog, Rana pipiens. Trinitrotoluene entering the organism was readily biotransformed to aminodinitrotoluenes, whereas no transformation products were measured for RDX or DNAN. Uptake clearance rates were relatively slow and similar among compounds (1.32-2.19 L kg(-1) h(-1) ). Upon transfer to uncontaminated water, elimination rate was very fast, resulting in the prediction of fast time to approach steady state (5 h or less) and short elimination half-lives (1.2 h or less). A preliminary bioconcentration factor of 0.25 L kg(-1) was determined for the insensitive munitions compound 3-nitro-1,2,4-trizole-5-one (NTO) indicating negligible bioaccumulative potential. Because of the rapid elimination rate for explosives, tadpoles inhabiting contaminated areas are expected to experience harmful effects only if under constant exposure conditions given that body burdens can rapidly depurate preventing tissue concentrations from persisting at levels that may cause detrimental biological effects. © 2014 SETAC.
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
Diaz-Badillo, Alvaro; Bolling, Bethany G; Perez-Ramirez, Gerardo; Moore, Chester G; Martinez-Munoz, Jorge P; Padilla-Viveros, America A; Camacho-Nuez, Minerva; Diaz-Perez, Alfonso; Beaty, Barry J; Munoz, Maria de Lourdes
2011-05-09
Culex spp. mosquitoes are considered to be the most important vectors of West Nile virus (WNV) detected in at least 34 species of mosquitoes in the United States. In North America, Culex pipiens pipiens, Culex pipiens quinquefasciatus, and Culex tarsalis are all competent vectors of WNV, which is considered to be enzootic in the United States and has also been detected in equines and birds in many states of Mexico and in humans in Nuevo Leon. There is potential for WNV to be introduced into Mexico City by various means including infected mosquitoes on airplanes, migrating birds, ground transportation and infected humans. Little is known of the geographic distribution of Culex pipiens complex mosquitoes and hybrids in Mexico City. Culex pipiens pipiens preferentially feed on avian hosts; Culex pipiens quinquefasciatus have historically been considered to prefer mammalian hosts; and hybrids of these two species could theoretically serve as bridge vectors to transmit WNV from avian hosts to humans and other mammalian hosts. In order to address the potential of WNV being introduced into Mexico City, we have determined the identity and spatial distribution of Culex pipiens complex mosquitoes and their hybrids. Mosquito larvae collected from 103 sites throughout Mexico City during 2004-2005 were identified as Culex, Culiseta or Ochlerotatus by morphological analysis. Within the genus Culex, specimens were further identified as Culex tarsalis or as belonging to the Culex pipiens complex. Members of the Culex pipiens complex were separated by measuring the ratio of the dorsal and ventral arms (DV/D ratio) of the male genitalia and also by using diagnostic primers designed for the Ace.2 gene. Culex pipiens quinquefasciatus was the most abundant form collected. Important WNV vectors species, Cx. p. pipiens, Cx. p. quinquefasciatus and Cx. tarsalis, are all present in Mexico City. Hybrids of Cx. p. pipiens and Cx. p. quinquefasciatus were also collected and identified. The
Ankley, Gerald T; Diamond, Stephen A; Tietge, Joseph E; Holcombe, Gary W; Jensen, Kathleen M; Defoe, David L; Peterson, Ryan
2002-07-01
A number of environmental stressors have been hypothesized as responsible for recent increases in limb malformations in several species of North American amphibians. The purpose of this study was to generate dose-response data suitable for assessing the potential role of solar ultraviolet (UV) radiation in causing limb malformations in a species in which this phenomenon seemingly is particularly prevalent, the northern leopard frog (Rana pipiens). Frogs were exposed from early embryonic stages through complete metamorphosis to varying natural sunlight regimes, including unaltered (100%) sunlight, sunlight subjected to neutral density filtration to achieve relative intensities of 85%, 75%, 65%, 50%, and 25% of unaltered sunlight, and sunlight filtered with glass or acrylamide to attenuate, respectively, the UVB (290-320 nm) and UVB plus UVA (290-380 nm) portions of the spectrum. The experiments were conducted in a controlled setting, with continual monitoring of UVB, UVA, and visible light to support a robust exposure assessment. Full sunlight caused approximately 50% mortality of the frogs during early larval development; no significant treatment-related mortality occurred under any of the other exposure regimes, including 100% sunlight with glass or acrylamide filtration. There was a dose-dependent (p < 0.0001) induction of hindlimb malformations in the frogs, with the percentage of affected animals ranging from about 97% under unaltered sunlight to 0% in the 25% neutral density treatment. Malformations were comprised mostly of missing or truncated digits, and generally were bilateral as well as symmetrical. Filtration of sunlight with either glass or acrylamide both significantly reduced the incidence of malformed limbs. The estimated sunlight dose resulting in a 50% limb malformation rate (ED50) was 63.5%. The limb ED50 values based on measured sunlight intensities corresponded to average daily doses of 4.5 and 100 Wh x m(-2) for UVB and UVA, respectively
First report of L1014F-kdr mutation in Culex pipiens complex from Morocco.
Bkhache, Meriem; Tmimi, Fatim-Zohra; Charafeddine, Omar; Faraj, Chafika; Failloux, Anna-Bella; Sarih, M'hammed
2016-12-16
Mosquitoes of the Culex pipiens complex, competent vectors for West Nile virus (WNV) and Rift Valley fever virus (RVFV) are widely targeted by insecticide treatments. The intensive application of chemical insecticides led to the development of resistance in many insects including Culex pipiens mosquitoes. The absence of data on resistance mechanisms in Morocco allow us to assess the levels of lambda-cyhalothrin resistance and the frequency of the mutated gene L1014F kdr in different forms of Cx. pipiens complex from three regions of Morocco. Mosquito adults were reared from immature stages collected in three different regions in Morocco (Tangier, Casablanca and Marrakech). Standard WHO insecticide susceptibility tests were conducted on adults emerged from collected larvae. Specimens were identified as belonging to the Culex pipiens complex using a multiplex PCR assay with diagnostic primers designed from the flanking region of microsatellite CQ11. Identified mosquitoes were then tested for the presence of the L1014F kdr mutation using PCR assay. Our results showed that 21% of the tested population has a resistance to lambda-cyhalothrin. The molecular identification of survivors shows that 43% belonged to the Cx. pipiens pipiens and only 9.5% to the Cx. pipiens molestus form. On the other hand, 416 specimens were screened for the L1014F kdr mutation. L1014F mutation was detected in different forms of Cx. pipiens in different sites. The frequency of L1014F mutation was similar between the Cx. pipiens pipiens form and hybrid form, while it was lower in the Cx. pipiens molestus form. The presence of the L1014F kdr allele was significantly associated with resistance to lambda-cyhalothrin in Cx. pipiens pipiens (P < 0.0001) and hybrid form (P < 0.0001). Resistance to lambda-cyhalothrin of Cx. pipiens populations appears to be largely due to the L1014F kdr mutation. To our knowledge, the frequencies of L1014F kdr mutation are examined for the first time in
Structure, Spatial and Temporal Distribution of the Culex pipiens Complex in Shanghai, China
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
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.
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...
Oviposition Activity Patterns of Culex pipiens and Culex restuans in Pennsylvania.
Stough, Jennifer E; Wallace, John R
2016-06-01
Culex pipiens and Cx. restuans are the main vectors of West Nile virus and the primary target species of surveillance and control programs in Pennsylvania. Performing adult control, specifically ultra-low volume (ULV) applications, at night during peak oviposition activity time(s) is necessary to control these species. In July and August of 2009, collections were made at 15-min intervals starting at sunset and continuing until 3 h after sunset to establish a more accurate timeline of Cx. pipiens and Cx. restuans oviposition flight activity. The highest numbers of Cx. pipiens and Cx. restuans were collected during the 15-30, 30-45, and 45-60 min postsunset time intervals (P < 0.05). Oviposition activity began to decrease after 60 min postsunset. These observations have identified a smaller oviposition activity period for Cx. pipiens and Cx. restuans than noted from other studies, thus potentially improving the timing of ULV operations to control these 2 vector species.
RANA CATESBEIANA (AMERICAN BULLFROG) DIET
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...
Gomes, Bruno; Sousa, Carla A; Novo, Maria T; Freitas, Ferdinando B; Alves, Ricardo; Côrte-Real, Ana R; Salgueiro, Patrícia; Donnelly, Martin J; Almeida, António P G; Pinto, João
2009-11-06
Culex pipiens L. is the most widespread mosquito vector in temperate regions. This species consists of two forms, denoted molestus and pipiens, that exhibit important behavioural and physiological differences. The evolutionary relationships and taxonomic status of these forms remain unclear. In northern European latitudes molestus and pipiens populations occupy different habitats (underground vs. aboveground), a separation that most likely promotes genetic isolation between forms. However, the same does not hold in southern Europe where both forms occur aboveground in sympatry. In these southern habitats, the extent of hybridisation and its impact on the extent of genetic divergence between forms under sympatric conditions has not been clarified. For this purpose, we have used phenotypic and genetic data to characterise Cx. pipiens collected aboveground in Portugal. Our aims were to determine levels of genetic differentiation and the degree of hybridisation between forms occurring in sympatry, and to relate these with both evolutionary and epidemiological tenets of this biological group. Autogeny and stenogamy was evaluated in the F1 progeny of 145 individual Cx. pipiens females. Bayesian clustering analysis based on the genotypes of 13 microsatellites revealed two distinct genetic clusters that were highly correlated with the alternative traits that define pipiens and molestus. Admixture analysis yielded hybrid rate estimates of 8-10%. Higher proportions of admixture were observed in pipiens individuals suggesting that more molestus genes are being introgressed into the pipiens form than the opposite. Both physiological/behavioural and genetic data provide evidence for the sympatric occurrence of molestus and pipiens forms of Cx. pipiens in the study area. In spite of the significant genetic differentiation between forms, hybridisation occurs at considerable levels. The observed pattern of asymmetric introgression probably relates to the different mating
2009-01-01
Background Culex pipiens L. is the most widespread mosquito vector in temperate regions. This species consists of two forms, denoted molestus and pipiens, that exhibit important behavioural and physiological differences. The evolutionary relationships and taxonomic status of these forms remain unclear. In northern European latitudes molestus and pipiens populations occupy different habitats (underground vs. aboveground), a separation that most likely promotes genetic isolation between forms. However, the same does not hold in southern Europe where both forms occur aboveground in sympatry. In these southern habitats, the extent of hybridisation and its impact on the extent of genetic divergence between forms under sympatric conditions has not been clarified. For this purpose, we have used phenotypic and genetic data to characterise Cx. pipiens collected aboveground in Portugal. Our aims were to determine levels of genetic differentiation and the degree of hybridisation between forms occurring in sympatry, and to relate these with both evolutionary and epidemiological tenets of this biological group. Results Autogeny and stenogamy was evaluated in the F1 progeny of 145 individual Cx. pipiens females. Bayesian clustering analysis based on the genotypes of 13 microsatellites revealed two distinct genetic clusters that were highly correlated with the alternative traits that define pipiens and molestus. Admixture analysis yielded hybrid rate estimates of 8-10%. Higher proportions of admixture were observed in pipiens individuals suggesting that more molestus genes are being introgressed into the pipiens form than the opposite. Conclusion Both physiological/behavioural and genetic data provide evidence for the sympatric occurrence of molestus and pipiens forms of Cx. pipiens in the study area. In spite of the significant genetic differentiation between forms, hybridisation occurs at considerable levels. The observed pattern of asymmetric introgression probably relates to
Trudeau, Vance L; Schueler, Frederick W; Navarro-Martin, Laia; Hamilton, Christine K; Bulaeva, Elizabeth; Bennett, Amanda; Fletcher, William; Taylor, Lisa
2013-02-25
Amphibian declines are now recognized globally. It is also well known that many anurans do not reproduce easily in captivity, especially when held over long periods, or if they require hibernation before breeding. A simple method to induce spawning and subsequent development of large numbers of healthy tadpoles is therefore required to meet research and conservation goals. The method is based on simultaneous injection of both female and male leopard frogs, Lithobates pipiens (formerly called Rana pipiens) with a cocktail of a gonadotropin-releasing hormone agonist (GnRH-A) and a dopamine antagonist. We call this the AMPHIPLEX method, which is derived from the combination of the words amphibian and amplexus. Following injection, the animals are thereby induced, and perform amplexus and natural fertilization under captive conditions. We tested combinations of a GnRH agonist with 2 different dopamine antagonists in L. pipiens in the breeding season. The combination of des-Gly(10), D-Ala(6), Pro-NHEt(9)-GnRH (0.4 micrograms/g body weight; GnRH-A) with metoclopramide hydrochloride (10 micrograms/g body weight; MET) or domperidone (DOM) were equally effective, producing 89% and 88% successful spawning, respectively. This yielded more than 44,000 eggs for the 16/18 females that ovulated in the GnRH-A+MET group, and more than 39,000 eggs for the 15/17 females that ovulated in the GnRH-A+DOM group. We further tested the GnRH-A+MET in frogs collected in the wild in late autumn and hibernated for a short period under laboratory conditions, and report a low spawning success (43%). However, GnRH-A priming 24 hours prior to injections of the GnRH-A+MET cocktail in animals hibernated for 5-6 weeks produced out-of-season spawning (89%) and fertilization (85%) comparable to those we observed for in-season spawning. Assessment of age and weight at metamorphosis indicated that L. pipiens tadpoles resulting from out-of-season spawning grew normally and metamorphosed successfully. We
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/♀.
2009-07-01
sediment protocol, respectively). The soil protocol evaluates impacts to adult salamanders and the sediment protocol evaluates impacts to larval tadpoles ...the sediment protocol evaluates impacts to larval tadpoles . When selecting appropriate receptors to derive ERA-based remedial goals, amphibians...Sediment tests are conducted with recently hatched tadpoles (i.e., Rana pipiens; Gosner Stages 17-20). Young tadpoles are placed in beakers
Culex pipiens Development Is Greatly Influenced by Native Bacteria and Exogenous Yeast
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
Repellent activity of five essential oils against Culex pipiens.
Erler, F; Ulug, I; Yalcinkaya, B
2006-12-01
Essential oils extracted from the seeds of anise (Pimpinella anisum), dried fruits of eucalyptus (Eucalyptus camaldulensis), dried foliage of mint (Mentha piperita) and basil (Ocimum basilicum) and fresh foliage of laurel (Laurus nobilis) were tested for their repellency against the adult females of Culex pipiens. All essential oils showed repellency in varying degrees, eucalyptus, basil and anise being the most active.
Bouhsira, E; Fysikopoulos, A; Franc, M
2009-08-01
A controlled clinical trial was carried out to assess the effectiveness of pyriprole, metaflumizone combined with amitraz, and fipronil-(S)-methoprene commercial spot-on products in preventing adult female Culex pipiens pipiens from feeding on dogs. Twenty-four beagle dogs were tested for their attractiveness to the mosquitoes and ranked accordingly to produce four groups of equivalent sensitivity to mosquitoes; six were treated with the pyriprole spot-on, six with the metaflumizone combined with amitraz spot-on, six with the fipronil-(S)-methoprene spot-on, and six were left untreated. All the dogs were challenged with 50 unfed adult female Culex in cages for one hour seven days before the treatment, and one and seven days after it. The mosquitoes were checked for blood feeding after one hour and for mortality 24 hours after they had been released into the cages. There was a significant reduction in the number of engorged mosquitoes only with the fipronil-(S)-methoprene product and there were significantly more dead mosquitoes with this product and the pyriprole product, but their effects were too small for them to be recommended for use in a dirofilariosis prevention programme.
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...
Beji, Marwa; Rhim, Adel; Roiz, David; Bouattour, Ali
2017-07-10
The Culex pipiens complex (Diptera: Culicidae) includes the most widespread mosquito species in the world. Members of this complex are the primary enzootic and epidemic vectors of the West Nile virus (genus Flavivirus) in several countries. The two recognized forms of Cx. pipiens (Linnaeus, 1758) - pipiens and molestus - exhibit behavioral and physiological differences. Natural populations of Cx. pipiens were investigated in several sites in Tunisia to evaluate the ecophysiological and molecular characteristics of their forms. The analysis showed the sympatric presence of Cx. pipiens forms and hybrids in all studied sites. Of all the tested larvae of Cx. pipiens, 33.5% were identified as pipiens, 30.8% were identified as molestus, and 35.6% were identified as hybrids. The molestus and hybrid forms were positively correlated with urban habitats and belowground sites while the pipiens form was positively correlated with rural habitats and aboveground sites. Autogeny was expressed in all types of habitats and breeding sites. By contrast with the microsatellite CQ11, the two molecular markers, ace-2 and cytb, did not allow differentiation between the Cx. pipiens forms. Our study shows the ubiquitous distribution and the plasticity of the different forms of Cx. pipiens in a wide range of ecological conditions. It suggests that the behavioral traits assigned to the forms of Cx. pipiens seem to be more flexible than previously assumed. Our analysis also proves that the microsatellite CQ11 remains an efficient tool for distinguishing between Cx. pipiens forms.
Cui, Jianxin; Li, Shujuan; Zhao, Ping; Zou, Fumin
2013-09-01
Culex pipiens pallens (L.) is the most common mosquito in houses of central and northern China. It is the primary vector of lymphatic filariasis and Japanese encephalitis. The flight range of mosquitoes is an important factor predicting the risk area of transmission of mosquito-borne pathogens to vertebrate hosts. The flight performance of Cx. pipiens pallens was measured with a 26-channel computer-monitored flight-mill system. We found that females had longer flight capability than males for total flight distance (TFD) and total flight duration (TFDr), and females flew faster than males based on mean flight velocity. No significant difference in flight capability was found between different age-groups in males. However, certain age-groups of females showed significant differences in TFDr and TFD. Specifically, TFD and TFDr tended to be shortest for 5- and 6-d-old females. These significant differences in flight capability between ages and genders provide insights to determine the size of operational area to achieve effective control of Cx. pipiens pallens and minimize the risk of the related mosquito-borne epidemic diseases of lymphatic filariasis and Japanese encephalitis.
Vogels, C B F; Göertz, G P; Pijlman, G P; Koenraadt, C J M
2017-12-01
In Europe, West Nile virus (WNV) outbreaks have been limited to southern and central European countries. However, competent mosquito vectors and susceptible bird hosts are present in northern Europe. Differences in temperature and vector competence of mosquito populations may explain the absence of WNV outbreaks in northern Europe. The aim of the present study was to directly compare vector competence of northern and southern European Culex pipiens (Cx. p.) pipiens mosquitoes for WNV across a gradient of temperatures. WNV infection and transmission rates were determined for two Cx. p. pipiens populations originating from The Netherlands and Italy, respectively. Mosquitoes were orally exposed by providing an infectious bloodmeal, or by injecting WNV (lineage 2) in the thorax, followed by 14-day incubation at 18, 23, or 28 °C. No differences in infection or transmission rates were found between the Cx. p. pipiens populations with both infection methods, but WNV transmission rates were significantly higher at temperatures above 18 °C. The absence of WNV outbreaks in northern Europe cannot be explained by differences in vector competence between Cx. p. pipiens populations originating from northern and southern Europe. This study suggests that low temperature is a key limiting factor for WNV transmission. © 2017 The Authors. Medical and Veterinary Entomology published by John Wiley & Sons Ltd on behalf of Royal Entomological Society.
Linthicum, Kenneth J
2012-12-01
Mosquitoes in the Culex pipiens Complex, including Culex pipiens, Cx. quinquefasciatus, and Cx. molestus, are important pest species and vectors of human and animal diseases throughout the world's tropical, temperate, and Holarctic regions. Diseases transmitted by member of the Pipiens Complex include: St. Louis encephalitis and West Nile virus in North America, West Nile virus on several other continents, Rift Valley fever in Africa, lymphatic filariasis caused by Wuchereria bancrofti in the tropics, and Dirofilaria immitis globally. Here and in the following 14 papers, 3 abstracts and a summary paper are the proceedings of a symposium that gathered many of the world's experts on the Pipiens Complex to explore the current state of knowledge of the taxa. Information presented at the symposium will improve our knowledge of important members of the complex and enhance our ability to conduct efficient surveillance and efficacious control strategies. A background on previous discussions on the Pipiens Complex, and a brief description of current symposium contributors and their topics are discussed.
Cetin, H; Yanikoglu, A; Cilek, J E
2006-06-01
The mosquito Culex pipiens L. is an important pest in urban and suburban areas in many parts of the world. Septic tanks are the most important habitats supporting the production of this species in the city of Antalya, southwestern Turkey. Diflubenzuron, in a 25% wettable powder (Du-dim 25 WP), and a 4% granular formulation (Du-dim 4 G) was evaluated against late 2nd to early 3rd instars of Cx. pipiens in single-family septic tanks. Both formulations were tested at 0.01, 0.02, and 0.03 mg (AI)/liter. The results indicated that both formulations applied at the rate of 0.02 and 0.03 mg (AI)/liter achieved 100% adult inhibition, at intervals of 7, 14, 21, and 28 days after treatment. Septic tanks treated with 0.01 mg (AI)/liter WP formulation resulted in complete (100%) adult inhibition through 14 days, whereas the G formulation gave the same effect through 21 days posttreatment at this rate.
Controlling Culex pipiens: antagonists are more efficient than a neonicotinoid insecticide.
Meyabeme Elono, Alvine Larissa; Foit, Kaarina; Duquesne, Sabine; Liess, Matthias
2018-06-01
Species vulnerability to pesticides depends on physiological sensitivity, the potential to recover, and the ecological context. We assessed the vulnerability of the mosquito Culex pipiens to a repeated treatment with thiacloprid in outdoor microcosms with and without antagonists (competitive and predatory invertebrates). Microcosms were treated repeatedly (three times) with thiacloprid at a concentration of 0.1, 1, or 10 µg/liter. In microcosms without antagonists, the abundance of Cx. pipiens larvae decreased moderately after the second and the third exposures to 10 µg/liter thiacloprid. In microcosms with antagonists, the abundance of Cx. pipiens larvae declined to approximately zero in the control group and the low concentration treatments during the five weeks of observation. By contrast, the abundance of Cx. pipiens larvae temporarily increased at 10 µg/liter thiacloprid after the second and third contamination. We explained this positive effect on the development of Cx. pipiens because of the decrease in competition due to the elimination of sensitive antagonists combined with the high recovery potential of Cx. pipiens. Based on these results, natural antagonists must be supported for the sustainable control of mosquitoes. © 2018 The Society for Vector Ecology.
A Pictorial Key for Culex pipiens Complex (Diptera: Culicidae) In Iran.
Dehghan, Hossein; Sadraei, Javid; Moosa-Kazemi, Seyed Hassan; Abolghasemi, Esmail; Solimani, Hassan; Jaffari Nodoshan, Ahmad; Najafi, Mohammad Hassan
2016-09-01
The aim of this study was to design pictorial key and taxonomic literature of Culex pipiens complex in Iran. 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. 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. It is necessary more studies on the behavior and genetic variations of Cx. pipiens complex in Iran.
COSTANZO, KATIE S.; MORMANN, KIMBERLY; JULIANO, STEVEN A.
2007-01-01
We tested for competitive advantage among larvae of Aedes albopictus (Skuse) and Culex pipiens L. in a laboratory experiment and determined the frequency and spatial and temporal patterns of co-occurrence in the field in East St. Louis, IL. In a laboratory competition experiment at multiple combined densities of Ae. albopictus and Cx. pipiens larvae, Ae. albopictus survivorship and developmental times were significantly affected by conspecific densities but not by Cx. pipiens densities. In contrast, Cx. pipiens survivorship and developmental times were significantly affected by both conspecific and Ae. albopictus densities. Per capita rate of increase (r′) for Ae. albopictus cohorts declined significantly due to density of conspecifics, but not density of Cx. pipiens. Interspecific competition between Ae. albopictus and Cx. pipiens under these laboratory conditions was strong and asymmetrical, with the effect of Ae. albopictus on Cx. pipiens much stronger than the reverse. In monthly samples from tire sites in East St. Louis, Ae. albopictus was highly seasonal, occurring in relatively low abundance from early May to July and increasing in abundance in August and September. Co-occurrence corresponded to the seasonality of Ae. albopictus, with Cx. pipiens encountering Ae. albopictus in more tires and at higher numbers within a tire, in August and September. Abundance of both species was high in residential areas and was unrelated to overstory cover, total nitrogen, and total phosphorus. Abundance of Ae. albopictus, but not of Cx. pipiens, was positively associated with conductivity. We expect Cx. pipiens to suffer from the effects of interspecific competition in tires in which it encounters Ae. albopictus. Interspecific competition between these species may be of both ecological and medical importance. PMID:16119544
Fritz, M L; Walker, E D; Miller, J R; Severson, D W; Dworkin, I
2015-06-01
Culex pipiens form pipiens and Cx. pipiens form molestus (Diptera: Culicidae) belong to a cosmopolitan taxonomic group known as the Pipiens Assemblage. Hybridization between these forms is thought to contribute to human transmission of West Nile virus (WNV) in North America. Complementary choice and no-choice landing assays were developed to examine host acceptance by North American Cx. pipiens in the laboratory. Populations collected from above- and below-ground sites in suburban Chicago were identified as forms pipiens and molestus using a polymerase chain reaction-based assay. Avian and human host acceptance was then quantified for the two populations, as well as for their hybrid and backcross offspring. No-choice tests were used to demonstrate that both the pipiens and molestus forms were capable of feeding on human and avian hosts. Choice tests were used to demonstrate that form pipiens females were strongly avian-seeking; an individual's probability of accepting the chick host was 85%. Form molestus females were more likely to accept the human host (87%). Rates of host acceptance by F1 and backcross progeny were intermediate to those of their parents. The results suggest that host preferences in Cx. pipiens are genetically determined, and that ongoing hybridization between above- and below-ground populations is an important contributor to epizootic transmission of WNV in North America. © 2015 The Royal Entomological Society.
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
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
Marini, Giovanni; Guzzetta, Giorgio; Baldacchino, Frederic; Arnoldi, Daniele; Montarsi, Fabrizio; Capelli, Gioia; Rizzoli, Annapaola; Merler, Stefano; Rosà, Roberto
2017-02-23
Aedes albopictus and Culex pipiens larvae reared in the same breeding site compete for resources, with an asymmetrical outcome that disadvantages only the latter species. The impact of these interactions on the overall ecology of these two species has not yet been assessed in the natural environment. In the present study, the temporal patterns of adult female mosquitoes from both species were analysed in north-eastern Italy, and substantial temporal shifts between abundance curves of Cx. pipiens and Ae. albopictus were observed in several sites. To understand which factors can drive the observed temporal shifts, we developed a mechanistic model that takes explicitly into account the effect of temperature on the development and survival of all mosquito stages. We also included into the model the effect of asymmetric interspecific competition, by adding a mortality term for Cx. pipiens larvae proportional to the larval abundance of Ae. albopictus within the same breeding site. Model calibration was performed through a Markov Chain Monte Carlo approach using weekly capture data collected in our study sites during 2014 and 2015. In almost half of observation sites, temporal shifts were due to competition, with an early decline of Cx. pipiens caused by the concurrent rise in abundance of its competitor, and this effect was enhanced by higher abundance of both species. We estimate that competition may reduce Cx. pipiens abundance in some sites by up to about 70%. However, in some cases temporal shifts can also be explained in the absence of competition between species resulting from a "temporal niche" effect, when the optimal fitness to environmental conditions for the two species are reached at different times of the year. Our findings demonstrate the importance of considering ecological interactions and, in particular, competition between mosquito species in temperate climates, with important implications for risk assessment of mosquito transmitted pathogens, as well
Calzolari, M; Bonilauri, P; Bellini, R; Becker, S; Dottori, M
2016-12-01
The Culex pipiens complex includes species with reported differences in vector competence for arthropod-borne viruses, many of which are of significant importance to human health such as the West Nile virus and the Sindbis virus. This group of mosquitoes is difficult to distinguish morphologically; particularly as adult females. In Europe, the two species of the complex, Culex pipiens Linnaeus 1758 and Culex torrentium Martini 1925, are often found sympatrically. With the aim to characterize the presence and spread of both species in the Emilia-Romagna region, Northern Italy, mosquitoes of the complex - collected during the West Nile virus surveillance plans - were tested by multiplex real-time PCR for the detection of the two species Cx. pipiens and Cx. torrentium. A total of 24 165 mosquitoes, collected between 2012 and 2014 from 105 sites, and sorted in 204 pools, were tested. All tested pools were found to be composed of Cx. pipiens, whereas Cx. torrentium was not detected. These results indicate a likely absence of Cx. torrentium mosquitoes within the surveyed territory, whereas Cx. pipiens is widely distributed in the area mentioned. This is in line with previous reports, which describe a pre-alpine distribution of Cx. torrentium in Italy. © 2016 The Royal Entomological Society.
Morphology of the caudal spinal cord in Rana (Ranidae) and Xenopus (Pipidae) tadpoles.
Nishikawa, K; Wassersug, R
1988-03-08
Using a variety of neuroanatomical and histological techniques, we compare the spinal cord and peripheral nerve distribution in the tails of larvae from Xenopus laevis and three species of Rana. The relatively large, postsacral spinal cord of Xenopus contains abundant motoneurons and their axons. Spinal nerves exit from the spinal cord in a regular array, one nerve per myotome, from the cervical region to near the end of the tail. Somata of motoneurons innervating caudal myotomes are found along the entire length of the tail. In contrast, the caudal cord of Rana is reduced to a filum terminale consisting of little more than an ependymal tube; spinal nerves to all caudal myotomes leave the cord in the sacral region and reach their motor targets via a cauda equina and caudal plexus. Motoneuron cell bodies innervating caudal myotomes are found only in the sacral region. The Rana larval pattern is similar to that of adult frogs and mammals, whereas the Xenopus larval pattern is more like that of salamanders and reptiles. These gross neuroanatomical differences are not due to differences in the size or developmental stage of the tadpoles, but instead are associated with differences in the swimming behavior of the larvae. The presence of motoneurons in the caudal spinal cord of Xenopus may provide local intermyotomal control within the tail; the elongated topography of the cord appears to permit finer, rostral-to-caudal regulation of neuromuscular activity. The Rana spinal cord, on the other hand--with motoneurons clustered anteriorly--may produce concurrent firing of adjacent ipsilateral myotomes, but at the expense of fine intermyotomal regulation. The fact that nerves in the tail of Xenopus enter and exit from the spinal cord locally, as opposed to far anteriorly as in Rana, means that for tadpoles of the same size, reflex arc lengths are many times shorter in Xenopus.
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
Bionomics and control of Culex pipiens fatigans Wied. in Ceylon
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
Vinogradova, E B
2003-01-01
Seasonal patterns of oviposition in a synanthropic unautogenous population of Culex pipiens pipiens mosquitoes from 69 suburban settlements (60 degrees 30' N, 30 degrees E') of the Leningrad province were studied during the period 1998-2002 years. The numbers of egg rafts laid in six artificial pools (barrels) were counted every day; altogether 692 rafts were collected. The general patterns of oviposition activity were similar for all studied years, in spite of their differences in the summer temperature regimes. The first peak of oviposition was observed during the second decade of July, the second peak--during the third decade of July--the first decade of August. The first and second summer generations of mosquitoes were responsible for these oviposition peaks, whereas the third generation completely entered the reproductive diapause. Thus, the oviposition activity was successfully used for populational monitoring of C. p. pipiens, that was for the first time recommended by Madder and co-authors (1980), taking into consideration the simplicity and economy of this technique. The differences in the attractiveness of distinct reservoirs for oviposition were recorded. The females preferred the barrels located on open sun-lit space and waters with organic pollution settled by mosquito larvae. Windly and rainy weather and also low (below 10 degrees C) night temperatures suppressed mosquito oviposition.
Zittra, Carina; Flechl, Eva; Kothmayer, Michael; Vitecek, Simon; Rossiter, Heidemarie; Zechmeister, Thomas; Fuehrer, Hans-Peter
2016-04-11
Culex pipiens complex taxa differ in behaviour, ecophysiology and epidemiologic importance. Despite their epidemiologic significance, information on genetic diversity, occurrence and seasonal and spatial distribution patterns of the Cx. pipiens complex is still insufficient. Assessment of seasonal and spatial distribution patterns of Culex pipiens forms and their congener Cx. torrentium is crucial for the understanding of their vector-pathogen dynamics. Female mosquitoes were trapped from April-October 2014 twice a month for a 24-h time period with BG-sentinel traps at 24 sampling sites in eastern Austria, using carbon dioxide as attractant. Ecological forms of Cx. pipiens s.l. and their hybrids were differentiated using the CQ11 locus, and Cx. pipiens forms and their congener Cx. torrentium using the ACE-2 gene. Differential exploitation of ecological niches by Cx. pipiens forms and Cx. torrentium was analysed using likelihood ratio tests. Possible effects of environmental parameters on these taxa were tested using PERMANOVA based on distance matrices and, if significant, were modelled in nMDS ordination space to estimate non-linear relationships. For this study, 1476 Culex spp. were sampled. Culex pipiens f. pipiens representing 87.33 % of the total catch was most abundant, followed by hybrids of both forms (5.62 %), Cx. torrentium (3.79 %) and Cx. pipiens f. molestus (3.25 %). Differences in proportional abundances were found between land cover classes. Ecological parameters affecting seasonal and spatial distribution of these taxa in eastern Austria are precipitation duration, air temperature, sunlight and the interaction term of precipitation amount and the Danube water level, which can be interpreted as a proxy for breeding habitat availability. The Cx. pipiens complex of eastern Austria comprises both ecologically different forms, the mainly ornithophilic form pipiens and the mainly mammalophilic and anthropophilic form molestus. Heterogeneous agricultural
Asymmetrical effects of introduced Rana catesbeiana on native ranid frogs in Oregon, USA
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.
Asymmetrical Effects of Introduced Bullfrogs (Rana catesbeiana) on Native Ranid Frogs in Oregon
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.
A regional survey of malformed frogs in Minnesota (USA) (Minnesota malformed frogs).
Vandenlangenberg, Susan M; Canfield, Jeffrey T; Magner, Joseph A
2003-02-01
In late 1995, school children discovered malformed frogs in a south central Minnesota pond. Press coverage resulted in numerous citizen reports of frog malformation across Minnesota in 1996. After some initial site investigation, 3 affected frog sites and 4 nearby reference sites were selected for more detailed evaluation. Field biologists made 89 visits to study sites beginning spring 1997 through fall 1999 to examine the number and type of frog malformations. Over 5,100 Leopard frogs (Rana pipiens) were captured and examined at all study sites. Water elevations and associated littoral inundation were recorded from 1997-2000. Results indicate that malformation occurred at all study sites above historical background levels. Rana pipiens malformation across all sites over three seasons averaged 7.9% and ranged from 0 to 7% at reference sites and 4 to 23% at affected sites. At one northern Minnesota site, mink frog (Rana septentrionalis) malformation was 75% in 1998. A site characteristic common to the most affected sites was an elastic zone of littoral inundation. Climate driven hydrologic variation likely influenced water depth and associated breeding locations.
The influence of clinostat rotation on the fertilized amphibian egg.
NASA Technical Reports Server (NTRS)
Tremor, J. W.; Souza, K. A.
1972-01-01
Study in which unrestrained, fertilized eggs of Rana pipiens and Xenopus laevis were rotated in a plane parallel to the normal gravity vector. In R. pipiens rotation at 1/4 rpm for five days at 18 C produced a significantly increased number of commonly occurring abnormalities. Rotation at 1/15, 1/8, 1, 2, 5 and 10 rpm did not significantly affect normal development. X. laevis eggs reacted similarly. R. pipiens eggs were most sensitive to rotation at 1/4 rpm when exposure was initiated before first cleavage. Mixing of intracellular constituents apparently occurred only at 1/4 rpm in R. pipiens (of the clinostat speeds studied), and may have been the cause of the increased abnormality observed at this rate.
Di Luca, Marco; Toma, Luciano; Boccolini, Daniela; Severini, Francesco; La Rosa, Giuseppe; Minelli, Giada; Bongiorno, Gioia; Montarsi, Fabrizio; Arnoldi, Daniele; Capelli, Gioia; Rizzoli, Annapaola; Romi, Roberto
2016-01-01
Mosquitoes in the Culex pipiens complex are considered to be involved in the transmission of a range of pathogens, including West Nile virus (WNV). Although its taxonomic status is still debated, the complex includes species, both globally distributed or with a more limited distribution, morphologically similar and characterised by different physiological and behavioural traits, which affect their ability as vectors. In many European countries, Cx. pipiens and its sibling species Culex torrentium occur in sympatry, exhibiting similar bionomic and morphological characters, but only Cx. pipiens appears to play a vector role in WNV transmission. This species consists of two biotypes, pipiens and molestus, which can interbreed when in sympatry, and their hybrids can act as WNV-bridge vectors, due to intermediate ecological features. Considering the yearly WNV outbreaks since 2008 and given the morphological difficulties in recognising species and biotypes, our aim was to molecularly identify and characterised Cx. pipiens and Cx. torrentium in Italy, using recently developed molecular assays. Culex torrentium was not detected; as in other European countries, the pipiens and molestus biotypes were widely found in sympatry with hybrids in most environments. The UPGMA cluster analysis applied to CQ11 genotypic frequencies mainly revealed two groups of Cx. pipiens populations that differed in ecological features. The high propensity of the molestus biotype to exist in hypogean environments, where the habitat's physical characteristics hinder and preclude the gene flow, was shown. These results confirmed the CQ11 assay as a reliable diagnostic method, consistent with the ecological and physiological aspects of the populations analysed. Since the assessment of the actual role of three biotypes in the WNV circulation remains a crucial point to be elucidated, this extensive molecular screening of Cx. pipiens populations can provide new insights into the ecology of the species
2005-04-01
Mountain juniper (Juniperus scopulorum), laurel leaf willow (Salix pentandra), Japanese lilac tree (Syringa reticulata), Amur chokecherry (Prunus maackii...Western painted turtle (Chrysemys picta belli), Northern leopard frog (Rana pipiens), Western chorus frog (Pseudacris triseriata), American toad (Bufo
Gardner, Allison M; Allan, Brian F; Frisbie, Lauren A; Muturi, Ephantus J
2015-06-16
Exotic invasive plants alter the structure and function of native ecosystems and may influence the distribution and abundance of arthropod disease vectors by modifying habitat quality. This study investigated how invasive plants alter the ecology of Culex pipiens, an important vector of West Nile virus (WNV) in northeastern and midwestern regions of the United States. Field and laboratory experiments were conducted to test the hypothesis that three native leaf species (Rubus allegheniensis, blackberry; Sambucus canadensis, elderberry; and Amelanchier laevis, serviceberry), and three exotic invasive leaf species (Lonicera maackii, Amur honeysuckle; Elaeagnus umbellata, autumn olive; and Rosa multiflora, multiflora rose) alter Cx. pipiens oviposition site selection, emergence rates, development time, and adult body size. The relative abundance of seven bacterial phyla in infusions of the six leaf species also was determined using quantitative real-time polymerase chain reaction to test the hypothesis that variation in emergence, development, and oviposition site selection is correlated to differences in the diversity and abundance of bacteria associated with different leaf species, important determinants of nutrient quality and availability for mosquito larvae. Leaf detritus from invasive honeysuckle and autumn olive yielded significantly higher adult emergence rates compared to detritus from the remaining leaf species and honeysuckle alleviated the negative effects of intraspecific competition on adult emergence. Conversely, leaves of native blackberry acted as an ecological trap, generating high oviposition but low emergence rates. Variation in bacterial flora associated with different leaf species may explain this asymmetrical production of mosquitoes: emergence rates and oviposition rates were positively correlated to bacterial abundance and diversity, respectively. We conclude that the displacement of native understory plant species by certain invasive shrubs
Ding, Yan-Mei; Hu, Yin; Yu, Bao-Ting; Mo, Xiao-Chang; Mo, Jian-Chu
2016-11-01
Mosquito adults usually need to obtain sugar from floral nectaries and damaged fruits/seed pods to replenish their energy reserves. The newly developed attractive toxic sugar baits have been successfully applied in controlling various mosquito species outdoors. However, the attraction of Culex pipiens pallens to different fruit-based sugar baits remains unknown. In the present study, we selected nine common fruit species, prepared the fruit-based sugar solutions, and investigated the attractiveness of different sugar baits to newly emerged Cx. pipiens pallens in the laboratory. The results showed that when tested against the 5% brown sugar solution, all the sugar baits were significantly attractive to both females and males. When tested together in the mesh-covered cage, there was a significant difference on the attractiveness between different fruit-based sugar baits. The most attractive fruit species included Broussonetia papyrifera, Cucumis melo, C. melo var. saccharinus, Amygdalus persica and Pyrus bretschneideri, and their seed pods could be potentially used as ingredients in ATSB for controlling mosquitoes outdoors. Copyright © 2016 Elsevier B.V. All rights reserved.
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
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.
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. This article is copyright of The Authors, 2016.
Cardo, M V; Rubio, A; Junges, M T; Vezzani, D; Carbajo, A E
2018-02-01
The mosquitoes of the Culex pipiens complex, hereafter referred to as the Pipiens Assemblage, are vectors of arbovirus of worldwide concern including West Nile and St. Louis encephalitis. Given their distinct eco-physiology and vectorial capacity, accurate specimen identification and insight in the environmental drivers of their distribution are essential for the understanding of disease transmission patterns. Using a PCR-based identification protocol, we characterized the spatial distribution of Cx. pipiens, Cx. quinquefasciatus and their hybrids developing in used tyres located within the overlapping region in South America as a function of different estimators of the urbanisation gradient. Out of 84 samples collected from tyre piles of 20 sites, we identified 369 larvae which corresponded predominantly to Cx. quinquefasciatus (76.4% of immatures) all along the gradient but more frequent at the urban end. Cx. pipiens (21.4%) was more conspicuous at the low urbanised end but was also present in highly urbanised sites, whereas hybrids were collected in very low numbers (2.2%). The urbanisation estimator best associated with the heterogeneous occurrence of the Pipiens Assemblage members was the proportion of impervious surface 1km around each tyre pile, which explained 41.7% of the variability in the data, followed closely by the distance to the Capital City (38.3%). Cumulative annual precipitation, population number in a 1km radius around each pile and distance to the de la Plata River were significantly associated with the distribution of the Pipiens Assemblage at lower explanation percentages (20-23%). A thorough understanding of the ecological basis and environmental associations of the distribution of Pipiens Assemblage members will enable forecasting population trends in changing environments to develop effective control measures for mosquitoes and the diseases they transmit. Copyright © 2017 Elsevier B.V. All rights reserved.
Krida, G; Rhim, A; Daaboub, J; Failloux, A-B; Bouattour, A
2015-06-01
Physiological and molecular characteristics of natural populations of Culex pipiens Linnaeus, 1758 (Diptera: Culicidae) were investigated to elucidate how this species is potentially involved in the transmission of West Nile virus in Tunisia. A total of 215 Cx. pipiens females from 11 breeding habitats were analysed in the laboratory to estimate autogeny and stenogamy rates. They were tested individually for the locus CQ11 to distinguish between the two Cx. pipiens forms, pipiens and molestus. All tested Cx. pipiens populations were stenogamous. Females from underground breeding sites were all autogeneous, whereas females from above-ground habitats were mostly anautogeneous. Of all the females tested, 59.7% were identified as pipiens, 22.4% as molestus, and 17.9% as hybrid pipiens/molestus. Furthermore, both Cx. pipiens forms and their hybrids were found to co-occur in sympatry in all sites. The results of this study represent the first evidence that both Cx. pipiens forms and their hybrids are present in Tunisia. Because hybrids able to act as bridge vectors are present in all studied habitats, Tunisia can be considered to have a high degree of receptivity for the establishment of West Nile virus zoonotic cycles. © 2015 The Royal Entomological Society.
Amara Korba, Raouf; Alayat, Moufida Saoucen; Bouiba, Lazhari; Boudrissa, Abdelkarim; Bouslama, Zihad; Boukraa, Slimane; Francis, Frederic; Failloux, Anna-Bella; Boubidi, Saïd Chaouki
2016-08-17
We investigated the ecological differentiation of two members of the Culex pipiens complex, Cx. p. pipiens form pipiens and Cx. p. pipiens form molestus in three sites, El-Kala, M'Sila and Tinerkouk in Algeria. These two forms are the most widespread mosquito vectors in temperate regions exhibiting important behavioural and physiological differences. Nevertheless, this group of potential vectors has been poorly studied, particularly in North Africa. Ten larval populations of Cx. p. pipiens were sampled from various above- and underground habitats in three zones representing the three bioclimatic regions in Algeria. The reproduction characteristics were also investigated in the laboratory to define the rates of autogeny and stenogamy. Identification of Cx. p. pipiens members present in Algeria was achieved using a molecular analysis with the microsatellite CQ11 locus. We detected larvae of Cx. p. pipiens in all areas suggesting that the species is a ubiquitous mosquito well adapted to various environments. To our knowledge, this study provides the first molecular evidence of the presence of the Cx. p. pipiens form molestus and hybrids (molestus/pipiens) in Algeria with a high proportion of molestus form (48.3 %) in comparison with hybrids (36.8 %) and pipiens form (14.9 %). Some unexpected correlations between the proportion of forms pipiens, molestus and hybrids, and mosquito biological characteristics were observed suggesting some epigenetic effects controlling Cx. p. pipiens mating and reproduction. Consequences for pathogen transmission are discussed.
Di Luca, Marco; Toma, Luciano; Boccolini, Daniela; Severini, Francesco; La Rosa, Giuseppe; Minelli, Giada; Bongiorno, Gioia; Montarsi, Fabrizio; Arnoldi, Daniele; Capelli, Gioia; Rizzoli, Annapaola; Romi, Roberto
2016-01-01
Mosquitoes in the Culex pipiens complex are considered to be involved in the transmission of a range of pathogens, including West Nile virus (WNV). Although its taxonomic status is still debated, the complex includes species, both globally distributed or with a more limited distribution, morphologically similar and characterised by different physiological and behavioural traits, which affect their ability as vectors. In many European countries, Cx. pipiens and its sibling species Culex torrentium occur in sympatry, exhibiting similar bionomic and morphological characters, but only Cx. pipiens appears to play a vector role in WNV transmission. This species consists of two biotypes, pipiens and molestus, which can interbreed when in sympatry, and their hybrids can act as WNV-bridge vectors, due to intermediate ecological features. Considering the yearly WNV outbreaks since 2008 and given the morphological difficulties in recognising species and biotypes, our aim was to molecularly identify and characterised Cx. pipiens and Cx. torrentium in Italy, using recently developed molecular assays. Culex torrentium was not detected; as in other European countries, the pipiens and molestus biotypes were widely found in sympatry with hybrids in most environments. The UPGMA cluster analysis applied to CQ11 genotypic frequencies mainly revealed two groups of Cx. pipiens populations that differed in ecological features. The high propensity of the molestus biotype to exist in hypogean environments, where the habitat’s physical characteristics hinder and preclude the gene flow, was shown. These results confirmed the CQ11 assay as a reliable diagnostic method, consistent with the ecological and physiological aspects of the populations analysed. Since the assessment of the actual role of three biotypes in the WNV circulation remains a crucial point to be elucidated, this extensive molecular screening of Cx. pipiens populations can provide new insights into the ecology of the species
Influence of anuran prey on the condition and distribution of Rana muscosa in the Sierra Nevada
K.L. Pope; K.R. Matthews
2002-01-01
Mountain yellow-legged frogs (Rana muscosa) at high elevations of the Sierra Nevada must obtain enough food during summer to survive 7â9 winter months when their aquatic habitats are frozen and food is presumably unavailable. Adults of R. muscosa prey on a variety of organisms, including aquatic and terrestrial invertebrates and...
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.
A 3-D QSAR-BASED IDENTIFICATION ALGORITHM FOR POTENTIAL ESTROGEN RECEPTOR LIGANDS
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...
Farajollahi, Ary; Fonseca, Dina M; Kramer, Laura D; Marm Kilpatrick, A
2011-10-01
The transmission of vector-borne pathogens is greatly influenced by the ecology of their vector, which is in turn shaped by genetic ancestry, the environment, and the hosts that are fed on. One group of vectors, the mosquitoes in the Culex pipiens complex, play key roles in the transmission of a range of pathogens including several viruses such as West Nile and St. Louis encephalitis viruses, avian malaria (Plasmodium spp.), and filarial worms. The Cx. pipiens complex includes Culex pipiens pipiens with two forms, pipiens and molestus, Culex pipiens pallens, Culex quinquefasciatus, Culex australicus, and Culex globocoxitus. While several members of the complex have limited geographic distributions, Cx. pipienspipiens and Cx. quinquefasciatus are found in all known urban and sub-urban temperate and tropical regions, respectively, across the world, where they are often principal disease vectors. In addition, hybrids are common in areas of overlap. Although gaps in our knowledge still remain, the advent of genetic tools has greatly enhanced our understanding of the history of speciation, domestication, dispersal, and hybridization. We review the taxonomy, genetics, evolution, behavior, and ecology of members of the Cx. pipiens complex and their role in the transmission of medically important pathogens. The adaptation of Cx. pipiens complex mosquitoes to human-altered environments led to their global distribution through dispersal via humans and, combined with their mixed feeding patterns on birds and mammals (including humans), increased the transmission of several avian pathogens to humans. We highlight several unanswered questions that will increase our ability to control diseases transmitted by these mosquitoes. Copyright © 2011 Elsevier B.V. All rights reserved.
Tabari, Mohaddeseh Abouhosseini; Youssefi, Mohammad Reza; Esfandiari, Aryan; Benelli, Giovanni
2017-10-01
Insect vectors are responsible for spreading devastating parasites and pathogens. A large number of botanicals have been suggested for eco-friendly control programs against mosquito vectors, and some of them are aromatic plants. Pelargonium roseum, a species belonging to the Geraniaceae family, due to its pleasant rose-like odor may represent a suitable candidate as mosquito repellent and/or larvicide. In this research, we evaluated the toxicity of the essential oil from P. roseum and its major constituents against the West Nile and filariasis vector Culex pipiens. The chemical composition of P. roseum essential oil was analyzed by gas chromatography-mass spectroscopy. Major constituents were citronellol (35.9%), geraniol (18.5%), and linalool (5.72%). The bioactivity of P. roseum essential oil and its three major compounds on larvae and egg rafts of Cx. pipiens was evaluated. The essential oil had a significant toxic effect on larvae and egg rafts of Cx. pipiens, with 50% lethal concentration (LC 50 ) values of 5.49 and 0.45μg/mL, respectively. Major constituents, geraniol, citronellol and linalool resulted in LC 50 values of 6.86, 7.64 and 14.87μg/mL on larvae, and 0.8, 0.67 and 1.27μg/mL on egg rafts. Essential oil and two of its constituents, citronellol and geraniol showed moderate knock-down on Cx. pipiens adults. Overall, the present investigation revealed that the major components of P. roseum and specially the whole essential oil could be helpful in developing novel and safe mosquito control tools and also offer an environmentally safe and cheap tool for reducing Cx. pipiens mosquito populations. Copyright © 2017. Published by Elsevier Ltd.
Fonseca, Dina M.; Smith, Julie L.; Kim, Heung-Chol; Mogi, Motoyoshi
2009-01-01
The Culex pipiens complex in Asia includes a temperate subspecies, Culex pipiens pallens, of uncertain taxonomic status. The shape of the male genitalia suggests it is a hybrid between Cx. pipiens and Cx. quinquefasciatus. We studied populations of Cx. p. pallens in Japan, Korea, and China and compared them to local populations of Cx. quinquefasciatus and Cx. p. pipiens. We examined variation in a nuclear intron in the acetylcholinesterase-2 gene [ACE] and eight microsatellite loci. We found a distinct microsatellite signature for Cx. p. pallens indicating restricted gene flow between Eastern and Western populations of Cx. pipiens, supporting the existence of two subspecies. Furthermore, a multilocus genotype analysis revealed current hybridization between Cx. p. pallens and Cx. quinquefasciatus in southern Japan, Republic of Korea, and China but not in Hokkaido, in northern Japan. Surprisingly, however, we found that the sex-linked ACE locus in chromosome I has introgressed asymmetrically through the males such that all male Cx. p. pallens have a copy of the Cx. quinquefasciatus ACE locus. This result highlights some of the potential consequences of hybridization between local and introduced species to disease transmission worldwide. PMID:19584006
Cetin, Huseyin; Yanikoglu, Atila; Cilek, James E
2005-06-01
The naturally-derived insecticide spinosad (Conserve SC) was evaluated against larval Culex pipiens L. (Diptera: Culicidae) under laboratory and field conditions in Antalya, Turkey. Laboratory bioassays showed that the 24 h LC50 and LC90 against late 3rd and early 4th instars were estimated at 0.027 and 0.111 parts per million, respectively, while adult emergence was eliminated at concentrations above 0.06 ppm. Larval mortality from septic tanks that were treated with spinosad at rates of 25, 50, 100, and 200 g ai/ha ranged between 22 to 78% 1 day after application. At 7 days post-treatment, larval mortality ranged from 2 to 50% and at 14 days mortality was <10% for all treatments. Larval bioassays of the water from those septic tanks treated at 100 and 200 g ai/ha resulted in an elimination of Cx. pipiens larvae 7 days after treatment. After this time, larval reduction declined to 79 and 83%, respectively, 14 days after treatment. Larval reduction in septic tanks treated at the two lowest rates (i.e. 25 and 50 g ai/ha) ranged from 14 to 74% during the 14-day study. These results indicated that spinosad can be considered an effective larvicide for treatment of septic tanks against Cx. pipiens.
Lõhmus, Mare; Lindström, Anders; Björklund, Mats
2012-01-01
Species in the Culex pipiens complex are common almost all over the world and represent important vectors for many serious zoonotic diseases. Even if, at the moment, many of the pathogens potentially transmitted by Cx. pipiens are not a problem in northern Europe, they may, with increasing temperatures and changing ecosystems caused by climate change, move northward in the future. Therefore, the question whether or not the Cx. pipiens populations in northern Europe will be competent vectors for them is of high importance. One way to estimate the similarity and the rate of contact between European Cx. pipiens populations is to look at the gene exchange between these populations. To test the genetic diversity and degree of differentiation between European Cx. pipiens populations, we used eight microsatellite markers in 10 mosquito populations originating from northern, central, and southern Europe. We found that three of the analyzed populations were very different from the rest of the populations and they also greatly differed from each other. When these three populations were removed, the variance among the rest of the populations was low, suggesting an extensive historic gene flow between many European Cx. pipiens populations. This suggests that infectious diseases spread by this species may not be associated with a certain vector genotype but rather with suitable environmental conditions. Consequently, we would expect these pathogens to disperse northward with favorable climatic parameters.
Toxicity of some plant extracts against vector of lymphatic filariasis, Culex pipiens.
Hasaballah, Ahmed I
2015-04-01
Many insecticides are generally used as larvicides to control Culex pipiens, vector of lymphatic filariasis. This study was undertaken to evaluate the larvicidal activity of some potential larvicidal plants extracts against C. pipiens larvae. The toxic effects of both ethanolic and petroleum ether plant extracts were evaluated under laboratory conditions against 3rd instar larvae of C. pipiens. Forty ethanolic and petroleum ether extracts of 10 plants namely Echinochloa stagninum, Phragmites australis, Eichhornia crassipes, Rhizophora mucronata, Cichorium intybus, Ocimum basilicum, Origanum majorana, Azadirachta indica, Rosmarinus officinalis and Nigella sativa. On the basis of LC50, the toxic effect of the plant extracts tested varied depending on the plant species, part, solvent used in extraction and the extract concentrations. The petroleum ether extraction was more effective against mosquito as compared with ethanolic extraction. The most effective plant extract was A. indica followed by Ph. australis, N. sativa, C. intybus, R. officinalis, O. basilicum, O. majorana, E. stagninum, Rh. Mucronata and E. crassipes.
Huang, Shaoming; Hamer, Gabriel L; Molaei, Goudarz; Walker, Edward D; Goldberg, Tony L; Kitron, Uriel D; Andreadis, Theodore G
2009-12-01
Mosquitoes of the Culex pipiens complex are important vectors of West Nile virus in the United States. We examined the genetic variations of Cx. pipiens mosquitoes from Chicago, Illinois that were determined to be principally ornithophilic but exhibited a relatively higher inclination for mammalian hosts including humans. Microsatellite analysis of 10 polymorphic markers was performed on 346 engorged Cx. pipiens specimens with identified avian or mammalian blood meals. Our results indicated that there were no significant differences in allelic richness, the pattern of conformity to Hardy-Weinberg equilibrium, and linkage disequilibrium, nor was there overall genetic differentiation between specimens with avian- and mammalian-derived blood meals. However, Cx. pipiens form pipiens with mammalian- (including human-) derived blood meals had significantly higher ancestry (p < 0.001) and proportion of hybrids (p < 0.01) from the Cx. pipiens form molestus (population from New York City) than did those with avian-derived blood meals. By contrast, there were no significant differences in the ancestry (p > 0.05) and the proportion of hybrids (p > 0.05) from Cx. quinquefasciatus (population from Harris Country, Texas). No temporal genetic variation was detected in accordance with the observation that there was no shift in blood feeding from birds to mammals. The results of this study in conjunction with regional host-feeding behavior suggest that the probability of genetic ancestry from Cx. pipiens f. molestus may predispose mosquitoes to feed more readily on mammals; however, the genetic mechanisms are unknown.
Wolbachia diversity and cytoplasmic incompatibility patterns in Culex pipiens populations in Turkey.
Altinli, Mine; Gunay, Filiz; Alten, Bulent; Weill, Mylene; Sicard, Mathieu
2018-03-20
Wolbachia are maternally transmitted bacteria that can manipulate their hosts' reproduction causing cytoplasmic incompatibility (CI). CI is a sperm-egg incompatibility resulting in embryonic death. Due to this sterilising effect on mosquitoes, Wolbachia are considered for vector control strategies. Important vectors for arboviruses, filarial nematodes and avian malaria, mosquitoes of Culex pipiens complex are suitable for Wolbachia-based vector control. They are infected with Wolbachia wPip strains belonging to five genetically distinct groups (wPip-I to V) within the Wolbachia B supergroup. CI properties of wPip strongly correlate with this genetic diversity: mosquitoes infected with wPip strains from a different wPip group are more likely to be incompatible with each other. Turkey is a critical spot for vector-borne diseases due to its unique geographical position as a natural bridge between Asia, Europe and Africa. However, general wPip diversity, distribution and CI patterns in natural Cx. pipiens (s.l.) populations in the region are unknown. In this study, we first identified wPip diversity in Turkish Cx. pipiens (s.l.) populations, by assigning them to one of the five groups within wPip (wPip-Ito V). We further investigated CI properties between different wPip strains from this region. We showed a wPip fixation in Cx. pipiens (s.l.) populations in Turkey by analysing 753 samples from 59 sampling sites. Three wPip groups were detected in the region: wPip-I, wPip-II and wPip-IV. The most dominant group was wPip-II. While wPip-IV was restricted to only two locations, wPip-I and wPip-II had wider distributions. Individuals infected with wPip-II were found co-existing with individuals infected with wPip-I or wPip-IV in some sampling sites. Two mosquito isofemale lines harbouring either a wPip-I or a wPip-II strain were established from a population in northwestern Turkey. Reciprocal crosses between these lines showed that they were fully compatible with each
ASYMMETRICAL EFFECTS OF INTRODUCED BULLFROGS (RANA CATESBEIANA) ON NATIVE RANID FROGS IN OREGON
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...
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.
Ferguson, Laura V; Kirk Hillier, N; Smith, Todd G
2013-12-01
Hepatozoon species are heteroxenous parasites that commonly infect the blood of vertebrates and various organs of arthropods. Despite their ubiquity, little is known about how these parasites affect host phenotype, including whether or not these parasites induce changes in hosts to increase transmission success. The objectives of this research were to investigate influences of the frog blood parasite Hepatozoon clamatae and the snake blood parasite Hepatozoon sipedon on host-seeking and host-choice behaviour of the mosquitoes Culex territans and Culex pipiens, respectively. During development of H. sipedon in C. pipiens, significantly fewer infected mosquitoes fed on uninfected snakes compared to uninfected mosquitoes. When H. sipedon was mature in C. pipiens, the number of infected and uninfected C. pipiens that fed on snakes was not significantly different. Higher numbers of mosquitoes fed on naturally infected snakes and frogs compared to laboratory-reared, uninfected control animals. However, experiments using only laboratory-raised frogs revealed that infection did not significantly affect host choice by C. territans. Behaviour of C. pipiens in the presence of H. sipedon may increase transmission success of the parasite and provide the first evidence of phenotypic changes in the invertebrate host of Hepatozoon parasites.
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
1993-01-01
Cut fibers from Rana temporaria and Rana pipiens (striation spacing, 3.9-4.2 microns) were mounted in a double Vaseline-gap chamber and studied at 14 degrees C. The Ca indicator purpurate-3,3' diacetic acid (PDAA) was introduced into the end pools and allowed to diffuse into the optical recording site. When the concentration at the site exceeded 2 mM, step depolarizations to 10 mV were applied and the [Ca] transient measured with PDAA was used to estimate Ca release from the sarcoplasmic reticulum (SR) (Baylor, S. M., W. K. Chandler, and M. W. Marshall. 1983. Journal of Physiology. 344:625-666). With depolarization, the rate of SR Ca release increased to an early peak and then rapidly decreased several-fold to a quasi-steady level. The total amount of Ca released from the SR at the time of peak rate of release appeared to be independent of SR Ca content, consistent with the idea that a single activated channel might pass, on average, a fixed number of ions, independent of the magnitude of the single channel flux. A possible explanation of this property is given in terms of locally induced Ca inactivation of Ca release. The solution in the end pools was then changed to one with PDAA plus fura-2. SR Ca release was estimated from the [Ca] transient, as before, and from the delta [Cafura-2] signal. On average, 2-3 mM fura-2 increased the quasi-steady level of the rate of SR Ca release by factors of 6.6 and 3.8, respectively, in three fibers from Rana temporaria and three fibers from Rana pipiens. The peak rate of release was increased in five of the six fibers but to a lesser extent than the quasi-steady level. In all fibers, the amplitude of the free [Ca] transient was markedly reduced. These increases in the rate of SR Ca release are consistent with the idea that Ca inactivation of Ca release develops during a step depolarization to 10 mV and that 2-3 mM fura-2 is able to reduce this inactivation by complexing Ca and thereby reducing free [Ca]. Once the concentration
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
Mavridis, Konstantinos; Fotakis, Emmanouil A; Kioulos, Ilias; Mpellou, Spiridoula; Konstantas, Spiros; Varela, Evangelia; Gewehr, Sandra; Diamantopoulos, Vasilis; Vontas, John
2018-06-01
During July-October 2017 a WNV outbreak took place in the Peloponnese, Southern Greece with five confirmed deaths. During routine monitoring survey in the Peloponnese, supported by the local Prefecture, we have confirmed the presence of all three Culex pipiens biotypes in the region, with a high percentage of Culex pipiens/molestus hybrids (37.0%) which are considered a highly competent vector of WNV. Kdr mutations related to pyrethroid resistance were found at relatively low levels (14.3% homozygosity) while no mosquitoes harboring the recently identified chitin synthase diflubenzuron-resistance mutations were detected in the region. As an immediate action, following the disease outbreak (within days), we collected a large number of mosquitoes using CO 2 CDC traps from the villages in the Argolis area of the Peloponnese, where high incidence of WNV human infections were reported. WNV lineage 2 was detected in 3 out of 47 Cx. pipiens mosquito pools (detection rate = 6.38%). The virus was not detected in any other mosquito species, such as Aedes albopictus, sampled from the region at the time of the disease outbreak. Our results show that detection of WNV lineage 2 in Cx. pipiens pools is spatially and chronologically associated with human clinical cases, thus implicating Cx. pipiens mosquitoes as the most likely WNV vector. The absence of diflubenzuron resistance mutations and the low frequency of pyrethroid (kdr) resistance mutations indicates the suitability of these insecticides for Cx. pipiens control, in the format of larvicides and/or residual spraying applications respectively, which was indeed the main (evidence based) response, following the disease outbreak. Copyright © 2018 Elsevier B.V. All rights reserved.
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.
The Role of Culex pipiens L. (Diptera: Culicidae) in Virus Transmission in Europe
Hernández-Triana, Luis M.; Medlock, Jolyon M.; Fooks, Anthony R.; Carpenter, Simon; Johnson, Nicholas
2018-01-01
Over the past three decades, a range of mosquito-borne viruses that threaten public and veterinary health have emerged or re-emerged in Europe. Mosquito surveillance activities have highlighted the Culex pipiens species complex as being critical for the maintenance of a number of these viruses. This species complex contains morphologically similar forms that exhibit variation in phenotypes that can influence the probability of virus transmission. Critical amongst these is the choice of host on which to feed, with different forms showing different feeding preferences. This influences the ability of the mosquito to vector viruses and facilitate transmission of viruses to humans and domestic animals. Biases towards blood-feeding on avian or mammalian hosts have been demonstrated for different Cx. pipiens ecoforms and emerging evidence of hybrid populations across Europe adds another level of complexity to virus transmission. A range of molecular methods based on DNA have been developed to enable discrimination between morphologically indistinguishable forms, although this remains an active area of research. This review provides a comprehensive overview of developments in the understanding of the ecology, behaviour and genetics of Cx. pipiens in Europe, and how this influences arbovirus transmission. PMID:29473903
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
Groen, Thomas A; L'Ambert, Gregory; Bellini, Romeo; Chaskopoulou, Alexandra; Petric, Dusan; Zgomba, Marija; Marrama, Laurence; Bicout, Dominique J
2017-10-26
Culex pipiens is the major vector of West Nile virus in Europe, and is causing frequent outbreaks throughout the southern part of the continent. Proper empirical modelling of the population dynamics of this species can help in understanding West Nile virus epidemiology, optimizing vector surveillance and mosquito control efforts. But modelling results may differ from place to place. In this study we look at which type of models and weather variables can be consistently used across different locations. Weekly mosquito trap collections from eight functional units located in France, Greece, Italy and Serbia for several years were combined. Additionally, rainfall, relative humidity and temperature were recorded. Correlations between lagged weather conditions and Cx. pipiens dynamics were analysed. Also seasonal autoregressive integrated moving-average (SARIMA) models were fitted to describe the temporal dynamics of Cx. pipiens and to check whether the weather variables could improve these models. Correlations were strongest between mean temperatures at short time lags, followed by relative humidity, most likely due to collinearity. Precipitation alone had weak correlations and inconsistent patterns across sites. SARIMA models could also make reasonable predictions, especially when longer time series of Cx. pipiens observations are available. Average temperature was a consistently good predictor across sites. When only short time series (~ < 4 years) of observations are available, average temperature can therefore be used to model Cx. pipiens dynamics. When longer time series (~ > 4 years) are available, SARIMAs can provide better statistical descriptions of Cx. pipiens dynamics, without the need for further weather variables. This suggests that density dependence is also an important determinant of Cx. pipiens dynamics.
Distortion product otoacoustic emissions provide clues to hearing mechanisms in the frog ear
NASA Astrophysics Data System (ADS)
Vassilakis, Pantelis N.; Meenderink, Sebastiaan W. F.; Narins, Peter M.
2004-12-01
2 f1-f2 and 2 f2-f1 distortion product otoacoustic emissions (DPOAEs) were recorded from both ears of male and female Rana pipiens pipiens and Rana catesbeiana. The input-output (I/O) curves obtained from the amphibian papilla (AP) of both frog species are analogous to I/O curves recorded from mammals suggesting that, similarly to the mammalian cochlea, there may be an amplification process present in the frog AP. DPOAE level dependence on L1-L2 is different from that in mammals and consistent with intermodulation distortion expectations. Therefore, if a mechanical structure in the frog inner ear is functioning analogously to the mammalian basilar membrane, it must be more broadly tuned. DPOAE audiograms were obtained for primary frequencies spanning the animals' hearing range and selected stimulus levels. The results confirm that DPOAEs are produced in both papillae, with R. catesbeiana producing stronger emissions than R. p. pipiens. Consistent with previously reported sexual dimorphism in the mammalian and anuran auditory systems, females of both species produce stronger emissions than males. Moreover, it appears that 2 f1-f2 in the frog is generated primarily at the DPOAE frequency place, while 2 f2-f1 is generated primarily at a frequency place around the primaries. Regardless of generation place, both emissions within the AP may be subject to the same filtering mechanism, possibly the tectorial membrane..
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...
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.
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
Brustolin, M; Talavera, S; Nuñez, A; Santamaría, C; Rivas, R; Pujol, N; Valle, M; Verdún, M; Brun, A; Pagès, N; Busquets, N
2017-12-01
Rift Valley fever (RVF) is a mosquito-borne disease caused by the Rift Valley fever virus (RVFV). Rift Valley fever affects a large number of species, including human, and has severe impact on public health and the economy, especially in African countries. The present study examined the vector competence of three different European mosquito species, Culex pipiens (Linnaeus, 1758) form molestus (Diptera: Culicidae), Culex pipiens hybrid form and Stegomyia albopicta (= Aedes albopictus) (Skuse, 1894) (Diptera: Culicidae). Mosquitoes were artificially fed with blood containing RVFV. Infection, disseminated infection and transmission efficiency were evaluated. This is the first study to assess the transmission efficiency of European mosquito species using a virulent RVFV strain. The virus disseminated in Cx. pipiens hybrid form and in S. albopicta. Moreover, infectious viral particles were isolated from saliva of both species, showing their RVFV transmission capacity. The presence of competent Cx. pipiens and S. albopicta in Spain indicates that an autochthonous outbreak of RVF may occur if the virus is introduced. These findings provide information that will help health authorities to set up efficient entomological surveillance and RVFV vector control programmes. © 2017 The Authors. Medical and Veterinary Entomology published by John Wiley & Sons Ltd on behalf of Royal Entomological Society.
Hybridization and population structure of the Culex pipiens complex in the islands of Macaronesia
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
Population size, survival, growth, and movements of Rana sierrae
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.
Shin, Ehyun; Park, Chan; Ahn, Young-Joon; Lee, Dong-Kyu; Chang, Kyu-Sik
2011-06-01
Culex pipiens molestus Forskal has been reported as a dominant species in underground structures of urban areas in the Republic of Korea (ROK) during all seasons and becomes bothersome to humans in late autumn and winter. Most Cx. pipiens molestus in septic tanks are controlled in the ROK using larvicides such as Bt and IGR. However, there are a number of problems associated with larvicides, such as high cost and requirement for frequent use. In the present work, a new control method for Cx. pipiens molestus in septic tanks by using mixtures of sucrose solution with insecticides was investigated. The insecticidal and repellent activities of ten insecticides were evaluated for best control of Cx. pipiens molestus in septic tanks. Firstly, differences in susceptibilities to insecticides were evaluated in topical assays by forced direct contact bioassay and in a screened wire cage by free direct contact bioassay. The difference in insecticide susceptibility in the mosquitoes was the result of repellency by the insecticides. In three septic tanks, the density of Culex mosquitoes was sharply reduced by a deltamethrin-sucrose solution kit. The results demonstrated the potential for mosquito control by deltamethrin-sucrose solution, and the study offers basic information related to mosquito control in septic tanks. Copyright © 2011 Society of Chemical Industry.
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.
Honnen, Ann-Christin; Monaghan, Michael T; Sharakhov, Igor
2017-09-01
Mosquitoes (Diptera, Culicidae) occur in natural, urban, and peri-urban areas throughout the globe. Although the characteristics of urban and peri-urban habitats differ from those of natural habitats in many ways (e.g., fragmentation, pollution, noise, and light), few studies have examined the population connectivity of mosquitoes in urban areas. To obtain an overview of the species composition, we sampled mosquitoes from 23 sites in and around the city of Berlin, Germany. Of 23 species, five occurred in urban, 10 in peri-urban, and 20 in rural areas. Culex pipiens Linnaeus (Diptera: Culicidae) was the most common species collected (75% of all individuals) and occurred in all habitats. Hence this species was selected to be analysed at 10 microsatellite markers. There was no significant differentiation (FST = 0.016, P = 0.9) or isolation by distance (P = 0.06) among Cx. pipiens populations along an urban-rural gradient. The only significant differences detected were between Cx. pipiens and a laboratory population of Cx. pipiens f. molestus (pairwise FST = 0.114-0.148, P ≤ 0.001 in all comparisons), suggesting that the markers chosen were suitable for the identification of population differentiation. Our results indicate that Cx. pipiens gene flow is widespread within and among urban, peri-urban, and rural areas and that urban habitat does not necessarily impede or enhance gene flow among these populations. © The Authors 2017. Published by Oxford University Press on behalf of Entomological Society of America.
Predation by Oregon spotted frogs (Rana pretiosa) on Western toads (Bufo boreas) in Oregon, USA
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.
Newman, Christina M; Anderson, Tavis K; Goldberg, Tony L
2016-01-01
Insect-specific flaviviruses (ISFVs) commonly infect vectors of mosquito-borne arboviruses. To investigate whether infection with an ISFV might affect mosquito flight behavior, we quantified flight behavior in Culex pipiens L. naturally infected with Culex flavivirus (CxFV). We observed a significant reduction in the scotophase (dark hours) flight activity of CxFV-positive mosquitoes relative to CxFV-negative mosquitoes, but only a marginal reduction in photophase (light hours) flight activity, and no change in the circadian pattern of flight activity. These results suggest that CxFV infection alters the flight activity of naturally infected Cx. pipiens most dramatically when these vectors are likely to be host seeking and may therefore affect the transmission of medically important arboviruses. © The Authors 2015. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.
USDA-ARS?s Scientific Manuscript database
Mosquitoes in the Culex pipiens complex, Cx. p. pipiens L. (Cpp) and Cx. p. quinquefasciatus Say (Cpq), are morphologically similar and important vectors of the West Nile and St. Louis Encephalitis viruses in the US. They hybridize when found in sympatry, which could facilitate the transfer of adva...
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.
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
A new species of Rana from the Dabie Mountains in eastern China (Anura, Ranidae)
Wang, Chencheng; Qian, Lifu; Zhang, Chenling; Guo, Weibo; Pan, Tao; Wu, Jun; Wang, Hui; Zhang, Baowei
2017-01-01
Abstract A new species Rana dabieshanensis sp. n. is described from the Dabie Mountains in Anhui Province, China, based on morphological character differences and molecular analyses. The new species can be distinguished from its congeners by a combination of diagnostic characters. The results of phylogenetic analyses (based on 12s rRNA, 16s rRNA, ND2, Cyt b, RAG1, BDNF and Tyr) and genetic distances (based on Cyt b) indicate that the new species belongs to the Rana longicrus group, and is placed as the sister taxon to R. hanluica. PMID:29362536
The problem of the Culex pipiens complex in the South Pacific (including Australia)*
Dobrotworsky, N. V.
1967-01-01
There are three representatives of the Culex pipiens complex in the South Pacific. C. p. fatigans is the most common and most widely distributed subspecies; it is closely associated with man. The males can be readily distinguished by the structure of the phallosome of the terminalia. C. p. molestus is spread over the southern part of Australia and in Tasmania; it also is a domestic mosquito. Throughout its extensive range in Australia, it exhibits all the biological traits that distinguish it from C. p. pipiens. C. p. australicus is widely distributed over the mainland of Australia and in Tasmania. It is superficially similar to C. p. fatigans but can be distinguished from C. p. pallens by the structure of the phallosome. It is primarily a rural non-man-biting mosquito. C. p. australicus is probably a relatively ancient member of the Australian fauna that may have evolved in the southern temperate zone. ImagesFIG. 3FIG. 4FIG. 2 PMID:5300062
Meenderink, Sebastiaan W F; van Dijk, Pim
2004-06-01
The inner ear of frogs holds two papillae specialized in detecting airborne sound, the amphibian papilla (AP) and the basilar papilla (BP). We measured input-output (I/O) curves of distortion product otoacoustic emissions (DPOAEs) from both papillae, and compared their properties. As in other vertebrates, DPOAE I/O curves showed two distinct segments, separated by a notch or kneepoint. The slope of the low-level segment was conspicuously different between the AP and the BP. For DPOAE I/O curves from the AP, slopes were < or = 1 dB/dB, similar to what is found in mammals, birds and some lizards. For DPOAE I/O curves from the BP these slopes were much steeper (approximately 2 dB/dB). Slopes found at high stimulus levels were similar in the AP and the BP (approximately 2 dB/dB). This quantitative difference between the low-level slopes for DPOAEs from the AP and the BP may signify the involvement of different mechanisms in low-level DPOAE generation for the two papillae, respectively.
Fankhauser, Becky; Dumont, Pascal; Hunter, James S; McCall, John W; Kaufmann, Christian; Mathis, Alexander; Young, David R; Carroll, Scott P; McCall, Scott; Chester, S Theodore; Soll, Mark D
2015-01-30
Three laboratory studies were conducted to assess the repellent and insecticidal efficacy of a combination of fipronil and permethrin (Frontline Tri- Act/Frontect) against three mosquito species (Aedes albopictus, Aedes aegypti and Culex pipiens) on dogs. In each study, 16 healthy adult dogs were allocated to two groups. Eight dogs were treated with the new topical spot-on combination of fipronil and permethrin on Day 0 and the other eight dogs served as untreated controls. Each dog was exposed to mosquitoes on Days 1, 7, 14, 21 and 28 (and also on Day 35 in the A. aegypti study). After a 1-h exposure period, all mosquitoes were counted and categorized as live or dead and fed or non-fed. Live mosquitoes were kept in an insectary and observed for mortality counts 4, 24 and 48 h post-exposure (PE) for Aedes spp. and 24 and 48 h PE for C. pipiens. Repellency and insecticidal efficacies were defined as the percent reduction in the number of fed and live mosquitoes, respectively, in the treated group as compared to the untreated control group. Repellency against A. albopictus was ≥93.4% through Day 21 and 86.9% on Day 28. It was ≥91.0% through Day 35 against A. aegypti and ≥90.4% through Day 28 against C. pipiens. Insecticidal efficacy against A. albopictus was ≥97.1% at 24 h PE from Day 7 to Day 28. It was ≥98.0% for the first 3 weeks and still 75.7% on Day 35 against A. aegypti at 24 h PE. For C. pipiens, insecticidal efficacy ranged from 93.8% (Day 7) to 30.9% (Day 28) at 48 h PE. A single topical administration of the combination of fipronil and permethrin provides repellency against mosquitoes on dogs for at least 4 weeks. The product may therefore significantly reduce the potential for the transmission of vector-borne pathogens through the inhibition of mosquito feeding, as well as the discomfort associated with mosquito bites. Moreover, mosquito mortality was induced by contact with the treated dogs, which could aid in the control of mosquitoes, and
Predicting Culex pipiens/restuans population dynamics by interval lagged weather data
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
Transmission of Rift Valley fever virus from European-breed lambs to Culex pipiens mosquitoes.
Vloet, Rianka P M; Vogels, Chantal B F; Koenraadt, Constantianus J M; Pijlman, Gorben P; Eiden, Martin; Gonzales, Jose L; van Keulen, Lucien J M; Wichgers Schreur, Paul J; Kortekaas, Jeroen
2017-12-01
Rift Valley fever virus (RVFV) is a mosquito-borne bunyavirus of the genus Phlebovirus that is highly pathogenic to ruminants and humans. The disease is currently confined to Africa and the Arabian Peninsula, but globalization and climate change may facilitate introductions of the virus into currently unaffected areas via infected animals or mosquitoes. The consequences of such an introduction will depend on environmental factors, the availability of susceptible ruminants and the capacity of local mosquitoes to transmit the virus. We have previously demonstrated that lambs native to the Netherlands are highly susceptible to RVFV and we here report the vector competence of Culex (Cx.) pipiens, the most abundant and widespread mosquito species in the country. Vector competence was first determined after artificial blood feeding of laboratory-reared mosquitoes using the attenuated Clone 13 strain. Subsequently, experiments with wild-type RVFV and mosquitoes hatched from field-collected eggs were performed. Finally, the transmission of RVFV from viremic lambs to mosquitoes was studied. Artificial feeding experiments using Clone 13 demonstrated that indigenous, laboratory-reared Cx. pipiens mosquitoes are susceptible to RVFV and that the virus can be transmitted via their saliva. Experiments with wild-type RVFV and mosquitoes hatched from field-collected eggs confirmed the vector competence of Cx. pipiens mosquitoes from the Netherlands. To subsequently investigate transmission of the virus under more natural conditions, mosquitoes were allowed to feed on RVFV-infected lambs during the viremic period. We found that RVFV is efficiently transmitted from lambs to mosquitoes, although transmission was restricted to peak viremia. Interestingly, in the mosquito-exposed skin samples, replication of RVFV was detected in previously unrecognized target cells. We here report the vector competence of Cx. pipiens mosquitoes from the Netherlands for RVFV. Both laboratory
Fortuna, Claudia; Remoli, Maria Elena; Di Luca, Marco; Severini, Francesco; Toma, Luciano; Benedetti, Eleonora; Bucci, Paola; Montarsi, Fabrizio; Minelli, Giada; Boccolini, Daniela; Romi, Roberto; Ciufolini, Maria Grazia
2015-09-17
West Nile virus (WNV) is a vector-borne disease responsible for causing epidemics in many areas of the world. The virus is maintained in nature by an enzootic bird-mosquito-bird cycle and occasionally transmitted to other hosts, such as equines and humans. Culex species, in particular the ubiquitous species Culex pipiens is thought to play a major vector role both in enzootic and epizootic maintenance and transmission of WNV. Introduced in Europe in recent years, since 2008 WNV has been stably circulating mainly in the Northeastern regions of Italy, although sporadic equine and/or human cases, as well as WNV infected Cx. pipiens pools, have been recorded in other Italian areas. The scope of our study was to evaluate the potential competence of some Italian populations of Cx. pipiens to transmit WNV and to assess their ability for vertical transmission of the virus. For this purpose four Italian populations, from different areas, were experimentally infected. After the infectious blood meal, fed females were monitored for 32 days to determine the length of viral extrinsic incubation period. WNV titre of infected mosquitoes was evaluated both by quantitative Real Time PCR and viral titration by Plaque Forming Units/ml (PFU/mL) in VERO cells. Potential Infection, Dissemination, Transmission rates (IR, DR, TR) were assessed by detection of the virus in body, legs plus wings and saliva of the fed females, respectively. All tested populations were susceptible to the WNV infection. The viral presence in legs and wings demonstrated the ability of WNV to disseminate in the mosquitoes. Viral RNA was detected in the saliva of tested populations. No significant differences in TR values were observed among the four studied populations. The offspring of the Cx. pipiens infected females were WNV negative. Our study addressed an important issue in the knowledge on the complex WNV-vector relationships in Italy, indicating that all Italian Cx. pipiens populations tested exhibited
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
DOE Office of Scientific and Technical Information (OSTI.GOV)
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,more » and cation concentrations in water at amphibian localities were negatively correlated with elevation. Survival of wood frog embryos declined when exposed to aluminum concentrations.« less
Kothera, Linda; Godsey, Marvin S.; Doyle, Michael S.; Savage, Harry M.
2012-01-01
Mosquitoes such as those in the Culex pipiens complex are important vectors of disease. This study was conducted to genetically characterize Cx. pipiens complex populations in the state of Colorado, USA, and to determine the number of genetic clusters represented by the data. Thirteen populations located among four major river basins were sampled (n = 597 individuals) using a panel of 14 microsatellites. The lowest-elevation sites had the highest Expected Heterozygosity (HE) values (range 0.54–0.65). AMOVA results indicated the presence of statistically significant amounts of variation within each level when populations were analyzed as one group or when they were grouped either by river basin or by their position on the east or west side of the Rocky Mountains. Most pairwise FST values were significant via permutation test (range 0–0.10), with the highest values from comparisons with Lamar, in southeast CO. A neighbor joining tree based on Cavalli–Sforza and Edwards’s chord distances was consistent with the geographic locations of populations, as well as with the AMOVA results. There was a significant isolation by distance effect, and the cluster analysis resolved five groups. Individuals were also assayed with an additional microsatellite marker, Cxpq78, proposed to be monomorphic in Cx. pipiens but polymorphic in the closely related but biologically distinct species Cx. quinquefasciatus. Low frequencies (≤3%) of Cx. quinquefasciatus alleles for this marker were noted, and mostly confined to populations along the Interstate 25 corridor. Pueblo was distinct in that it had 10% Cx. quinquefasciatus alleles, mostly of one allele size. The degree of population genetic structure observed in this study is in contrast with that of Cx. tarsalis, the other major vector of WNV in the western U.S., and likely reflects the two species’ different dispersal strategies. PMID:23094068
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.
The phenotypic variability in Rana temporaria decreases in response to drying habitats.
Miramontes-Sequeiros, Luz Calia; Palanca-Castán, Nicolás; Caamaño-Chinchilla, Laura; Palanca-Soler, Antonio
2018-01-15
In this study, we evaluated the diversity of skin coloration as a proxy for phenotypic diversity. The European common frog (Rana temporaria) populations from the Southern slope of central Pyrenees lie at the limit of the species distribution in latitude and altitude. We analysed the relationship of skin color typology with different environmental variables and found a large decrease in skin type variety in frogs developing in temporary water bodies when compared to those developing in permanent water bodies. Our results show that our method can be used as a non-invasive way to study phenotypic diversity and suggest that adaptation to an early metamorphosis in a rapidly-drying habitat can have negative effects on adult phenotypic diversity. In light of these results, we argue that access to permanent water bodies is important to prevent loss of diversity in anuran populations and reduce their vulnerability to environmental impacts as well as pathogens. Copyright © 2017 Elsevier B.V. All rights reserved.
[Adaptive features of life history for space experiments seen in Rana tagoi tadpoles].
Yamashita, Masamichi; Kobayashi, Ken-ichiro; Kashiwagi, Akihiko; Shigenari, Makiko; Wassersug, Richard J; Naitoh, Tomio
2002-11-01
We propose to use the physiological responses of tadpoles in space to study the effects of gravity at the organismal level. The tadpoles we propose for study, Rana tagoi, have naturally transparent abdominal walls, permitting viscera, such as heart and intestine, to be easily observed. Rana tagoi is a mountainous species that lays eggs in water, typically at crevasses in rocky substrates. Its eggs are rich in yolk, and tadpoles can grow without being fed (except own feces). However, they will eat when presented food. Coprophagy in these tadpoles can be clearly seen when small amount of food is fed. Abdominal wall remains transparent until metamorphosis. Intestinal and cardiac activity are clearly visible and can be documented by video without any invasive procedures. Because of the limited requirements on the digestive system in this species, their intestine does not elongate much when the tadpoles grow up. Even in this species, periodical contractions occur in the intestine throughout the larval period. Space experiments using R. tagoi tadpoles are promising in terms of scientific merit and technical feasibility. With this species it is possible to study the effects of gravity on visceral physiology and metamorphosis. Rana tagoi tadpoles would be a good model for space experiments where only the basic minimum capability of life support function (e.g., temperature and oxygen control), could be provided.
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.
Kim, Soon-Il; Ahn, Young-Joon
2017-05-04
The yellow fever mosquito, Aedes aegypti, and the common house mosquito, Culex pipiens pallens, transmit dengue fever and West Nile virus diseases, respectively. This study was conducted to determine the toxicity of the three lignans (-)-asarinin, sesamin and (+)-xanthoxylol-γ,γ-dimethylallylether (XDA), and the alkaloid pellitorine from Zanthoxylum piperitum (Rutaceae) bark to third-instar larvae from insecticide-susceptible C. pipiens pallens and Ae. aegypti as well as wild C. pipiens pallens resistant to deltamethrin, cyfluthrin, fenthion, and temephos. The toxicities of all isolates were compared with those of mosquito larvicide temephos. LC 50 values for each species and their treatments were significantly different from one another when their 95% confidence intervals did not overlap. XDA was isolated from Z. piperitum as a new larvicidal principle. XDA (LC 50 , 0.27 and 0.24 mg/l) was 4, 53, and 144 times and 4, 100, and 117 times more toxic than pellitorine, sesamin, and asarinin toward larvae from susceptible C. pipiens pallens and Ae. aegypti, respectively. Overall, all the isolates were less toxic than temephos (LC 50 , 0.006 and 0.009 mg/l). These constituents did not differ in toxicity to larvae from the two Culex strains. The present finding indicates that the lignans and alkaloid and the insecticides do not share a common mode of larvicidal action or elicit cross-resistance. Naturally occurring Z. piperitum bark-derived compounds, particularly XDA, merit further study as potential mosquito larval control agents or as lead compounds for the control of insecticide-resistant mosquito populations.
2011-01-01
Background Insecticide resistance jeopardizes the control of mosquito populations and mosquito-borne disease control, which creates a major public health concern. Two-dimensional electrophoresis identified one protein segment with high sequence homology to part of Aedes aegypti iron-responsive element binding protein (IRE-BP). Method RT-PCR and RACE (rapid amplification of cDNA end) were used to clone a cDNA encoding full length IRE-BP 1. Real-time quantitative RT-PCR was used to evaluate the transcriptional level changes in the Cr-IRE strain Aedes aegypti compared to the susceptible strain of Cx. pipiens pallens. The expression profile of the gene was established in the mosquito life cycle. Methyl tritiated thymidine (3H-TdR) was used to observe the cypermethrin resistance changes in C6/36 cells containing the stably transfected IRE-BP 1 gene of Cx. pipiens pallens. Results The complete sequence of iron responsive element binding protein 1 (IRE-BP 1) has been cloned from the cypermethrin-resistant strain of Culex pipiens pallens (Cr-IRE strain). Quantitative RT-PCR analysis indicated that the IRE-BP 1 transcription level was 6.7 times higher in the Cr-IRE strain than in the susceptible strain of 4th instar larvae. The IRE-BP 1 expression was also found to be consistently higher throughout the life cycle of the Cr-IRE strain. A protein of predicted size 109.4 kDa has been detected by Western blotting in IRE-BP 1-transfected mosquito C6/36 cells. These IRE-BP 1-transfected cells also showed enhanced cypermethrin resistance compared to null-transfected or plasmid vector-transfected cells as determined by 3H-TdR incorporation. Conclusion IRE-BP 1 is expressed at higher levels in the Cr-IRE strain, and may confer some insecticide resistance in Cx. pipiens pallens. PMID:22075242
Hemoparasites in Oregon spotted frogs (Rana pretiosa) from central Oregon, USA.
Stenberg, Patricia L; Bowerman, William J
2008-04-01
Between 2001 and 2003, we screened blood smears of 156 Oregon spotted frogs (Rana pretiosa) from three populations in central Oregon for blood parasites. A Lankesterella sp. and a Trypanosoma sp. were detected in 31% and 35% of the frogs, respectively. Parasite loads were generally light, with Lankesterella sporozoites in 1-2% of erythrocytes, and extracellular trypanosomes were seen at rates of about one parasite per 200 fields of view at 1,000x. Little work has been published on hemoparasites of ranids in the western USA in the past 30 yr. Because of the recent taxonomic division of the Rana pretiosa complex, this may be the first published report of blood parasites for R. pretiosa sensu stricto. Both parasites reported here differed in morphologic features and morphometric comparisons from previous descriptions of anuran hemoparasites. Much work remains to sort out the taxonomy of hemoparasites among western USA ranids and to determine the ecological significance of these parasites; both tasks are important steps in understanding and managing these, and related, sensitive and threatened species.
Pillard, David A; Eck, William S; Johnson, Mark S; Packard, Stephanie
2017-11-01
New explosive formulations are being developed to be less sensitive to impact and inadvertent explosion, increasing safety for the warfighter. Since testing and training make environmental releases imminent, the toxicity of 3-nitro-1,2,4-triazol-5-one (NTO), a component of Insensitive Munitions eXplosive (IMX) formulations, was assessed in a one-generation study to the northern leopard frog (Lithobates ( = Rana) pipiens). Because NTO in water creates acidic conditions, acute studies were conducted with non-pH-adjusted NTO, while a long-term (70-d) study was conducted with neutralized NTO. In the acute study, 48-h and 7-d LC 50 s were ~250 mg NTO/L. In the long-term study, tadpoles were dead by day 2 in 11,350 mg/L NTO, and by day 63 in 8382 mg/L. The 70-d LC 50 was 3670 mg (neutralized) NTO/L. The number of organisms reaching complete metamorphosis was reduced by NTO; the lowest IC 25 was 1999 mg NTO/L for the Number Completing Metamorphosis. The NOECs for Time to Front Limb Eruption or Time to Metamorphosis were the same at 1346 mg/L. Histopathology did not significantly distinguish between NTO-exposed and unexposed animals, although possible effects on the density of spermatogonia in NTO-exposed males was suggested. The test data indicate that acute toxicity to ambient NTO can be attributed primarily to its acidic nature; relatively low chronic toxicity of neutralized NTO is due to delays in metamorphosis. The consequence from this latter observation may be ecologically significant as delays of even a few days could increase mortality through predation and/or loss of the aquatic medium in temporary water bodies.
Bigot, Diane; Atyame, Célestine M; Weill, Mylène; Justy, Fabienne
2018-01-01
Abstract In the global context of arboviral emergence, deep sequencing unlocks the discovery of new mosquito-borne viruses. Mosquitoes of the species Culex pipiens, C. torrentium, and C. hortensis were sampled from 22 locations worldwide for transcriptomic analyses. A virus discovery pipeline was used to analyze the dataset of 0.7 billion reads comprising 22 individual transcriptomes. Two closely related 6.8 kb viral genomes were identified in C. pipiens and named as Culex pipiens associated tunisia virus (CpATV) strains Ayed and Jedaida. The CpATV genome contained four ORFs. ORF1 possessed helicase and RNA-dependent RNA polymerase (RdRp) domains related to new viral sequences recently found mainly in dipterans. ORF2 and 4 contained a capsid protein domain showing strong homology with Virgaviridae plant viruses. ORF3 displayed similarities with eukaryotic Rhoptry domain and a merozoite surface protein (MSP7) domain only found in mosquito-transmitted Plasmodium, suggesting possible interactions between CpATV and vertebrate cells. Estimation of a strong purifying selection exerted on each ORFs and the presence of a polymorphism maintained in the coding region of ORF3 suggested that both CpATV sequences are genuine functional viruses. CpATV is part of an entirely new and highly diversified group of viruses recently found in insects, and that bears the genomic hallmarks of a new viral family. PMID:29340209
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
Russell, Danelle M; Goldberg, Caren S; Waits, Lisette P; Rosenblum, Erica Bree
2010-11-01
The pathogenic chytrid fungus Batrachochytrium dendrobatidis (Bd) is contributing to amphibian declines worldwide. Temperature plays an important role in both pathogen growth and host immune function, but little is known about seasonal dynamics of Bd infection in north temperate regions. Our objective was to increase understanding of Bd disease ecology by investigating patterns of Bd infection of Columbia spotted frogs Rana luteiventris across seasons, age classes, and sexes in north Idaho, USA. We collected skin swabs from 223 R. luteiventris in spring, summer, and fall 2009 at 7 ponds in the Palouse region and quantified Bd zoospores for each sample using quantitative PCR. Across seasons, Bd prevalence of adults was higher in summer than in spring or fall, suggesting that individuals may be clearing low-level infections over the summer. Among age classes, all but one late stage tadpole (Gosner stage 43-45) tested negative for Bd. Conversely, 100% of metamorphs tested positive for Bd and had the highest Bd loads of all age classes, suggesting they may be the most vulnerable age class. Adult R. luteiventris had high infection prevalence (> 60%) in all seasons, indicating that Bd infection is maintained within populations and that adults likely serve as disease reservoirs across seasons. Among adults, we also found weak evidence for females having higher infection prevalence than males. Further laboratory and field studies are needed to determine whether there are individual and population impacts from Bd on R. luteiventris and other amphibians in north Idaho.
Microsatellite primers for Culex pipiens quinquefasciatus, the vector of avian malaria in Hawaii
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.
Geographic variation in the response of Culex pipiens life history traits to temperature.
Ruybal, Jordan E; Kramer, Laura D; Kilpatrick, A Marm
2016-02-29
Climate change is predicted to alter the transmission of many vector-borne pathogens. The quantitative impact of climate change is usually estimated by measuring the temperature-performance relationships for a single population of vectors, and then mapping this relationship across a range of temperatures or locations. However, life history traits of different populations often differ significantly. Specifically, performance across a range of temperatures is likely to vary due to local adaptation to temperature and other factors. This variation can cause spatial variation in pathogen transmission and will influence the impact of climate change on the transmission of vector-borne pathogens. We quantified variation in life history traits for four populations of Culex pipiens (Linnaeus) mosquitoes. The populations were distributed along altitudinal and latitudinal gradients in the eastern United States that spanned ~3 °C in mean summer temperature, which is similar to the magnitude of global warming expected in the next 3-5 decades. We measured larval and adult survival, development rate, and biting rate at six temperatures between 16 and 35 °C, in a common garden experiment. Temperature had strong and consistent non-linear effects on all four life history traits for all four populations. Adult female development time decreased monotonically with increasing temperature, with the largest decrease at cold temperatures. Daily juvenile and adult female survival also decreased with increasing temperature, but the largest decrease occurred at higher temperatures. There was significant among-population variation in the thermal response curves for the four life history traits across the four populations, with larval survival, adult survival, and development rate varying up to 45, 79, and 84 % among populations, respectively. However, variation was not correlated with local temperatures and thus did not support the local thermal adaptation hypothesis. These results suggest
Boothe, Emily; Medeiros, Matthew C I; Kitron, Uriel D; Brawn, Jeffrey D; Ruiz, Marilyn O; Goldberg, Tony L; Walker, Edward D; Hamer, Gabriel L
2015-05-01
Multiple mosquito-borne parasites cocirculate in nature and potentially interact. To understand the community of parasites cocirculating with West Nile virus (WNV), we screened the bloodmeal content of Culex pipiens L. mosquitoes for three common types of hemoparasites. Blood-fed Cx. pipiens were collected from a WNV-epidemic area in suburban Chicago, IL, from May to September 2005 through 2010. DNA was extracted from dissected abdomens and subject to PCR and direct sequencing to identify the vertebrate host. RNA was extracted from the head or thorax and screened for WNV using quantitative reverse transcriptase PCR. Seventy-nine engorged females with avian host origin were screened using PCR and amplicon sequencing for filarioid nematodes, Haemosporida, and trypanosomatids. Filarioid nematodes were identified in 3.8% of the blooded abdomens, Plasmodium sp. in 8.9%, Haemoproteus in 31.6%, and Trypanosoma sp. in 6.3%. The sequences from these hemoparasite lineages were highly similar to sequences from birds in prior studies in suburban Chicago. Overall, 50.6% of blood-fed Culex pipiens contained hemoparasite DNA in their abdomen, presumably from current or prior bloodmeals. Additionally, we detected hemoparasite DNA in the blooded abdomen of three of 10 Cx. pipiens infected with WNV. © The Authors 2015. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.
Indirect effects of introduced trout on Cascades frogs (Rana cascadae) via shared aquatic prey
Maxwell B. Joseph; Jonah Piovia-Scott; Sharon P. Lawler; Karen L. Pope
2010-01-01
1. The introduction of trout to montane lakes has negatively affected amphibian populations across the western United States. In northern California’s Klamath–Siskiyou Mountains, introduced trout have diminished the distribution and abundance of a native ranid frog, Rana (=Lithobates)
Theoretical Series Elastic Element Length in Rana pipiens Sartorius Muscles
Matsumoto, Yorimi
1967-01-01
Assuming a two component system for the muscle, a series elastic element and a contractile component, the analyses of the isotonic and isometric data points were related to obtain the series elastic stiffness, dP/dls, from the relation, See PDF for Equation From the isometric data, dP/dt was obtained and shortening velocity, v, was a result of the isotonic experiments. Substituting (P 0 - P)/T for dP/dt and (P 0 - P)/(P + a) times b for v, dP/dls = (P + a) /bT, where P < P 0, and a, b are constants for any lengths l ≤ l 0 (Matsumoto, 1965). If the isometric tension and the shortening velocity are recorded for a given muscle length, l 0, although the series elastic, ls, and the contractile component, lc, are changing, the total muscle length, l 0 remains fixed and therefore the time constant, T. Integrating, See PDF for Equation the stress-strain relation for the series elastic element, See PDF for Equation is obtained; l sc0 - ls + l c0where l co equals the contractile component length for a muscle exerting a tension of P 0. For a given P/P 0, ls is uniquely determined and must be the same whether on the isotonic or isometric length-tension-time curve. In fact, a locus on one surface curve can be associated with the corresponding locus on the other. PMID:6033578
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
Foothill yellow-legged frog (Rana boylii) oviposition site choice at multiple spatial scales
Amy J. Lind; Hartwell H. Welsh; Clara A. Wheeler
2016-01-01
Studies of resource selection at multiple scales are critical to understanding ecological and evolutionary attributes of a species. We analyzed relative abundance, habitat use, and oviposition site selection of Foothill Yellow-Legged Frogs (Rana boylii) at 11 localities across two geographic regions in California (northern Coast Range and Sierra...
Mating strategy and breeding patterns of the foothill yellow-legged frog (Rana boylii)
Clara A. Wheeler; Hartwell H. Welsh Jr.
2008-01-01
The Foothill Yellow-legged Frog (Rana boylii) has declined across much of its native range in California. Improper stream management may lower egg mass survival and reduce the availability of suitable breeding habitats. We collected data during six breeding-seasons (2002-2007) along an unregulated stream in northwestern California. We monitored...
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
Bolling, Bethany G.; Olea-Popelka, Francisco J.; Eisen, Lars; Moore, Chester G.; Blair, Carol D.
2012-01-01
We established a laboratory colony of Culex pipiens mosquitoes from eggs collected in Colorado and discovered that mosquitoes in the colony are naturally infected with Culex flavivirus (CxFV), an insect-specific flavivirus. In this study we examined transmission dynamics of CxFV and effects of persistent CxFV infection on vector competence for West Nile virus (WNV). We found that vertical transmission is the primary mechanism for persistence of CxFV in Cx. pipiens, with venereal transmission potentially playing a minor role. Vector competence experiments indicated possible early suppression of WNV replication by persistent CxFV infection in Cx. pipiens. This is the first description of insect-specific flavivirus transmission dynamics in a naturally infected mosquito colony and the observation of delayed dissemination of superinfecting WNV suggests that the presence of CxFV may impact the intensity of enzootic transmission of WNV and the risk of human exposure to this important pathogen. PMID:22425062
Bolling, Bethany G; Olea-Popelka, Francisco J; Eisen, Lars; Moore, Chester G; Blair, Carol D
2012-06-05
We established a laboratory colony of Culex pipiens mosquitoes from eggs collected in Colorado and discovered that mosquitoes in the colony are naturally infected with Culex flavivirus (CxFV), an insect-specific flavivirus. In this study we examined transmission dynamics of CxFV and effects of persistent CxFV infection on vector competence for West Nile virus (WNV). We found that vertical transmission is the primary mechanism for persistence of CxFV in Cx. pipiens, with venereal transmission potentially playing a minor role. Vector competence experiments indicated possible early suppression of WNV replication by persistent CxFV infection in Cx. pipiens. This is the first description of insect-specific flavivirus transmission dynamics in a naturally infected mosquito colony and the observation of delayed dissemination of superinfecting WNV suggests that the presence of CxFV may impact the intensity of enzootic transmission of WNV and the risk of human exposure to this important pathogen. Copyright © 2012 Elsevier Inc. All rights reserved.
Responses of foothill yellow-legged frog (Rana boylii) larvae to an introduced predator
David J. Paoletti; Deanna H. Olson; Andrew R. Blaustein
2011-01-01
The consequences of species introductions into non-native habitats are a major cause for concern in the U.S. Of particular interest are the effects of predation by introduced fishes on native amphibian communities. We sought to determine whether Foothill Yellow-legged Frog (Rana boylii) larvae could recognize non-native Small mouth Bass (...
Yang, Bao-Tian; Zhou, Yu; Min, Mi-Sook; Matsui, Masafumi; Dong, Bing-Jun; Li, Pi-Peng; Fong, Jonathan J
2017-07-01
We investigated the species diversity and phylogeography of the Northeast Asian brown frogs allied to Rana dybowskii (the R. dybowskii species complex: R. dybowskii, R. pirica, and R. uenoi) using four mitochondrial and three nuclear loci. Phylogenetic analyses confirmed the existence of three distinct species in this complex; using extensive molecular data, we confirm the validity of Rana uenoi recognized as a distinct species, and infer R. dybowskii and R. pirica to be sister species. Also, we included populations from previously unsampled regions in Northeast China, and identified them to be R. dybowskii. While many species in Northeast Asia diverged due to Pleistocene glaciation, divergence-dating analyses inferred older, Miocene speciation in the R. dybowskii species complex. Ancestral area reconstruction identified the orogenic movement of the Changbai Mountain Range and the opening of the Sea of Japan/East Sea being major events influencing allopatric speciation. Copyright © 2017 Elsevier Inc. All rights reserved.
Changes in breeding phenology of eastern Ontario frogs over four decades.
Klaus, Samantha P; Lougheed, Stephen C
2013-04-01
Global climate change has been implicated in phenological shifts for a variety of taxa. Amphibian species in particular are sensitive to changes in their environment due to their biphasic life history and restricted reproductive requirements. Previous research has shown that not all temperate amphibian species respond similarly to the same suite of climatic or environmental cues, nor are individual species necessarily uniform in their responses across their range. We examined both the timing of spring emergence and calling phenology of eight anuran species in southeastern Ontario, Canada, using an approximately 40-year dataset of historical records of amphibian activity. Rana pipiens was the only species out of eight considered to emerge significantly earlier, by an estimated 22 days over four decades. Both R. pipiens and Bufo americanus have advanced initiation of calling over a four-decade span significantly earlier by an estimated 37.2 and 19.2 days, respectively. Rana sylvatica showed a trend toward earlier emergence by 19 days, whereas we did not detect changes in emergence phenology for the remaining five species. This significant shift in breeding behavior for two species correlates to significant regional increases in spring temperatures of an estimated 2.7-2.8°C overall over four decades. Our study suggests that local temperature increases have affected the timing of emergence and the onset of calling activity in some Ontario anuran species. Global decline or range shifts ultimately may be related to changes in reproductive behavior and timing mediated by shifting climate.
POPULATION STATUS AND DISTRIBUTION OF A DECIMATED AMPHIBIAN, THE RELICT LEOPARD FROG (RANA ONCA)
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...
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.
Site fidelity of the declining amphibian Rana sierrae (Sierra Nevada yellow-legged frog)
Kathleen Matthews; Haiganoush Preisler
2010-01-01
From 1997 to 2006, we used markârecapture models to estimate the site fidelity of 1250 Sierra Nevada yellow-legged frogs (Rana sierrae) in Kings Canyon National Park, California, USA, during their three main activity periods of overwintering, breeding, and feeding. To quantify site fidelity, the tendency to return to and reuse previously occupied...
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.
Toma, Luciano; Menegon, Michela; Romi, Roberto; De Matthaeis, Elvira; Montanari, Mario; Severini, Carlo
2011-01-01
Heavy and constant use of organophosphorus (OP) larvicides selected Culex pipiens L. resistant populations through two main mechanisms of genetic resistance, the increased activity of detoxifying esterase and the production of alterate acetylcholinesterase-1 (AChE1) by G119S mutation. The aim of this study was the assessment of the distribution of Cx. pipiens populations resistant to temephos and chlorpyrifos in the north-eastern regions of Italy and the occurrence of the insensitive AChE in these populations. Data describe the situation in the last years before European legislation prohibited the use of OP larvicides in mosquito control, up until 2007. For the first time a high level of OP resistance in the samples from Ravenna (182-fold, 80% A4/B4 or A5/B5 esterases and 38.3% Ester(5)), Emilia Romagna region, was detected; therefore, new data from the Veneto and Friuli Venezia Giulia regions were obtained and reinforced existing knowledge about resistance previously studied along the Adriatic coast. Nearby, in the Villa Verucchio locality, the highest (87.5%) AChE1R was found. Cx. pipiens resistance esterases A5/B5 and A4/B4 spread southward along the Adriatic coastal plain while OPs were being used in mosquito control, as confirmed by the first molecular screening of the AChE1 gene in these populations. Copyright © 2010 Society of Chemical Industry.
Liu, Hong-Mei; Cheng, Peng; Huang, Xiaodan; Dai, Yu-Hua; Wang, Hai-Fang; Liu, Li-Juan; Zhao, Yu-Qiang; Wang, Huai-Wei; Gong, Mao-Qing
2013-02-01
The present study aimed to investigate deltamethrin resistance in Culex pipiens pallens (C. pipiens pallens) mosquitoes and its correlation with knockdown resistance (kdr) mutations. In addition, mosquito‑resistance testing methods were analyzed. Using specific primers in polymerase chain reaction (PCR) and allele-specific (AS)-PCR, kdr gene sequences isolated from wild C. pipiens pallens mosquitoes were sequenced. Linear regression analysis was used to determine the correlation between the mutations and deltamethrin resistance. A kdr allelic gene was cloned and sequenced. Analysis of the DNA sequences revealed the presence of two point mutations at the L1014 residue in the IIS6 transmembrane segment of the voltage‑gated sodium channel (VGSC): L1014F, TTA→TTT, replacing a leucine (L) with a phenylalanine (F); L1014S, TTA→TCA, replacing leucine (L) with serine (S). Two alternative kdr-like mutations, L1014F and L1014S, were identified to be positively correlated with the deltamethrin-resistant phenotype. In addition a novel mutation, TCT, was identified in the VGSC of C. pipiens pallens. PCR and AS-PCR yielded consistent results with respect to mosquito resistance. However, the detection rate of PCR was higher than that of AS-PCR. Further studies are required to determine the specific resistance mechanism. PCR and AS-PCR demonstrated suitability for mosquito resistance field tests, however, the former method may be superior to the latter.
Liang, Yaping; Li, Xiuwei; Gu, Zumin; Qin, Peiwen; Ji, Mingshan
2015-02-16
The larvicidal activity of the crude petroleum ether, ethyl acetate, acetone, chloroform and ethanol extracts of Amorpha fruticosa seeds was individually assayed for toxicity against the early fourth-instar larva of the mosquito, Culex pipiens pallens after 24 h exposure. Of the tested extracts, the ethanol one exhibited the highest larvicidal activity (LC50 = 22.69 mg/L). Amorphigenin (8'-hydroxyrotenone), a rotenoid compound which exhibits a strong larvicidal activity with LC50 and LC90 values of 4.29 and 11.27 mg/L, respectively, was isolated from the ethanol extract by column chromatograpy. Its structure was elucidated by 1H-NMR, UV and IR spectral data. Furthermore, investigation of amorphigenin's effects on mitochondrial complex I activity and protein synthesis in C. pipiens pallens larvae reveals that amorphigenin decreases mitochondrial complex I activities to 65.73% at 10.45 μmol/L, compared to the control, when NADH were used as the substrate. Meanwhile, amorphigenin at 10.45 μmol/L also caused a 1.98-fold decrease in protein content, compared to the control larvae treated with acetone only.
Richards, Christopher T
2010-02-15
This study aimed to compare the swimming kinematics and hydrodynamics within and among aquatic and semi-aquatic/terrestrial frogs. High-speed video was used to obtain kinematics of the leg joints and feet as animals swam freely across their natural range of speeds. Blade element analysis was then used to model the hydrodynamic thrust as a function of foot kinematics. Two purely aquatic frogs, Xenopus laevis and Hymenochirus boettgeri, were compared with two semi-aquatic/terrestrial frogs, Rana pipiens and Bufo americanus. The four species performed similarly. Among swimming strokes, peak stroke velocity ranged from 3.3+/-1.1 to 20.9+/-2.5, from 6.8+/-2.1 to 28.6+/-3.7 and from 4.9+/-0.5 to 20.9+/-4.1 body lengths per second (BL s(-1)) in X. laevis, H. boettgeri and R. pipiens, respectively (means +/- s.d.; N=4 frogs for each). B. americanus swam much more slowly at 3.1+/-0.3 to 7.0+/-2.0 BL s(-1) (N=3 frogs). Time-varying joint kinematics patterns were superficially similar among species. Because foot kinematics result from the cumulative motion of joints proximal to the feet, small differences in time-varying joint kinematics among species resulted in species-specific foot kinematics (therefore hydrodynamics) patterns. To obtain a simple measure of the hydrodynamically useful motion of the foot, this study uses 'effective foot velocity' (EFV): a measure of the component of foot velocity along the axis of swimming. Resolving EFV into translational and rotational components allows predictions of species-specific propulsion strategies. Additionally, a novel kinematic analysis is presented here that enables the partitioning of translational and rotational foot velocity into velocity components contributed by extension at each individual limb joint. Data from the kinematics analysis show that R. pipiens and B. americanus translated their feet faster than their body moved forward, resulting in positive net translational EFV. Conversely, translational EFV was slower than
Pachka, Hammami; Annelise, Tran; Alan, Kemp; Power, Tshikae; Patrick, Kgori; Véronique, Chevalier; Janusz, Paweska; Ferran, Jori
2016-08-08
In Northern Botswana, rural communities, livestock, wildlife and large numbers of mosquitoes cohabitate around permanent waters of the Okavango Delta. As in other regions of sub-Saharan Africa, Rift Valley Fever (RVF) virus is known to circulate in that area among wild and domestic animals. However, the diversity and composition of potential RVF mosquito vectors in that area are unknown as well as the climatic and ecological drivers susceptible to affect their population dynamics. Using net traps baited with carbon dioxide, monthly mosquito catches were implemented over four sites surrounding cattle corrals at the northwestern border of the Okavango Delta between 2011 and 2012. The collected mosquito species were identified and analysed for the presence of RVF virus by molecular methods. In addition, a mechanistic model was developed to assess the qualitative influence of meteorological and environmental factors such as temperature, rainfall and flooding levels, on the population dynamics of the most abundant species detected (Culex pipiens). More than 25,000 mosquitoes from 32 different species were captured with an overabundance of Cx. pipiens (69,39 %), followed by Mansonia uniformis (20,67 %) and a very low detection of Aedes spp. (0.51 %). No RVF virus was detected in our mosquito pooled samples. The model fitted well the Cx. pipiens catching results (ρ = 0.94, P = 0.017). The spatial distribution of its abundance was well represented when using local rainfall and flooding measures (ρ = 1, P = 0.083). The global population dynamics were mainly influenced by temperature, but both rainfall and flooding presented a significant influence. The best and worst suitable periods for mosquito abundance were around March to May and June to October, respectively. Our study provides the first available data on the presence of potential RVF vectors that could contribute to the maintenance and dissemination of RVF virus in the Okavango Delta. Our model allowed us to
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
Morçiçek, Burçin; Taskin, Belgin Gocmen; Doğaç, Ersin; Doğaroğlu, Taylan; Taskin, Vatan
2018-06-01
Establishing reliable risk projection information about the distribution pattern of members of the Culex pipiens complex is of particular interest, as these mosquitoes are competent vectors for certain disease-causing pathogens. Wolbachia, a maternally inherited bacterial symbiont, are distributed in various arthropod species and can induce cytoplasmic incompatibility, i.e., reduced egg hatch, in certain crosses. It is being considered as a tool for population control of mosquito disease vectors. The Aegean region is characterized by highly populated, rural, and agricultural areas and is also on the route of the migratory birds. In this study, a fragment of the 658 bp of the mitochondrial cytochrome c oxidase subunit 1 (COI) gene, which includes the barcode region, was employed to differentiate Cx. pipiens complex species found in this region. Moreover, for the first time, the prevalence of Wolbachia endobacteria in these natural populations was examined using PCR amplification of a specific wsp gene. Our results revealed a widespread (more than 90%, n=121) presence of the highly efficient West Nile virus vector Cx. quinquefasciatus in the region. We also found that Wolbachia infection is widespread; the average prevalence was 62% in populations throughout the region. This study provided valuable information about the composition of Cx. pipiens complex mosquitoes and the prevalence of Wolbachia infection in these populations in the Aegean region. This information will be helpful in tracking mosquito-borne diseases and designing and implementing Wolbachia-based control strategies in the region. © 2018 The Society for Vector Ecology.
Al-Mekhlafi, Fahd A; Abutaha, Nael; Mashaly, Ashraf M A; Nasr, Fahd A; Ibrahim, Khalid E; Wadaan, Mohamed A
2017-05-01
Effects of methanol extracts of Xanthium strumarium on different cancer cell lines and on the mortality rates of Aedes caspius, Culex pipiens (Diptera: Culicidae) were investigated. Among the cell lines tested, the Jurkat cell line was the most sensitive to the methanol extract and ethyl acetate fraction, with reported LC 50 values of 50.18 and 48.73 μg/ml respectively. Conversely, methanol extracts were not that toxic to the A549 cell line though the toxicity increased on further purification. The percentage of growth inhibition was dose dependent for the methanol extract and ethyl acetate fraction. The ethyl acetate fraction showed higher toxicity to all cell lines tested when compared to the methanol extract. The results showed that methanol extracts of plant seeds caused 100% mortality of mosquito larvae at a concentration of 1000 μg/ml after 24 h of treatment. The LC 50 and LC 90 values of X. strumarium were found to be 531.07 and 905.95 μg/ml against Ae. caspius and 502.32 and 867.63 μg/ml against Cx. Pipiens, respectively. From the investigations, it was concluded that the crude extract of X. strumarium showed a weak potential for controlling the larval instars of Ae. caspius and Cx. pipiens . However, on further purification the extract lost the larvicidal activity. The ethyl acetate fraction showed higher toxicity to all cell lines tested when compared to the methanol extract. The ethyl acetate fraction investigated in this study appears to have a weak larvicidal activity but a promising cytotoxic activity. Future studies will include purification and investigation in further detail of the action of X. strumarium on Cancer Cell Lines and mosquitoes.
Bruner, M A; Shipman, P A; Rao, M; Bantle, J A
2002-09-01
This study assessed the effects of ambient UV light on the development of two native species of anurans, Rana blairi and Hyla chrysoscelis, during their normal breeding season in Oklahoma. Additionally, the effects of ambient UV light and water contaminated with landfill leachate in Rana blairi were examined. Embryos were collected from the field and distributed equally among replicates of four filter treatments of ambient UV light in experimental tubs filled with either FETAX solution or landfill leachate diluted to 25, 10, and 5% concentrations. Three endpoints (mortality, teratogenesis, and growth) were compared between filter treatments. By itself, UV-B caused no significant effects. Leachate at 10 and 25% concentrations caused 100% mortality across all filter treatments. There was a significant interaction between filter treatment and water toxicity at leachate concentrations of 5% for both malformation and growth. Increased UV-B exposure decreased the malformation rate and increased growth in the leachate treatments.
K.R. Matthews; K.L. Pope
1999-01-01
ABSTRACTS.âIn a high-elevation (3470 m) lake basin (upper Dusy Basin) in Kings Canyon National Park, California, we used radio transmitters on 24 mountain yellow-legged frogs (Rana muscosa) to gather basic information on their movement patterns. Rana muscosa have declined throughout their range in the Sierra Nevada and restoration plans require information on their...
Myofiber turnover is used to retrofit frog jaw muscles during metamorphosis.
Alley, K E
1989-01-01
Metamorphic reorganization of the head in anuran amphibians entails abrupt restructuring of the jaw complex as larval feeding structures are transformed into their adult configurations. In this morphometric study, light microscopy wa used to analyze the larval maturation and metamorphic transfiguration of the adductor jaw muscles in the leopard frog (Rana pipiens). Larval jaw muscles, first established during embryogenesis, continue to grow by fiber addition until prometamorphosis, stage XII. Thereafter, fiber number remains stable but additional muscle growth continues by hypertrophy of the individual fibers until metamorphic climax. During metamorphic stages XIX-XXIII, a complete involution of all larval myofibers occurs. Simultaneously, within the same muscle beds, a second wave of myogenesis produces myoblasts which are the precursors of adult jaw myofibers. New muscle fibers continue to be added to these muscles well after the completion of metamorphosis; however, the total duration of the postmetamorphic myogenic period has not been defined. These observations provide clear evidence that the entir population of primary myofibers used in larval oral activity disappears from the adductor muscle beds and is replaced by a second wave of myogenesis commencing during climax. These findings indicate that the adductor jaw muscles are prepared for adult feeding by a complicated cellular process that retrofits existing muscle beds with a completely new complement of myofibers.
Stephen F. Hale; Philip C. Rosen; James L. Jarchow; Gregory A. Bradley
2005-01-01
We conducted histological analyses on museum specimens collected 1975-1999 from 10 sites in Arizona and Sonora to test for the pathogenic chytrid fungus (Batrachochytrium dendrobatidis) in ranid frogs, focusing on the Tarahumara frog (Rana tarahumarae). During 1981-2000, frogs displaying disease signs were found in the field, and...
ELECTRIC IMPEDANCE OF THE FROG EGG
Cole, Kenneth S.; Guttman, Rita M.
1942-01-01
Electrical impedance measurements were made upon unfertilized and fertilized eggs of the leopard frog, Rana pipiens, over a frequency range of 0.05 to 10 kc. Average values of 170 ohm cm.2 were obtained for the plasma membrane resistance of the egg, 2.0 µf/cm.2 for the plasma membrane capacity, 86° for the phase angle of the membrane, and 570 ohm cm. for the specific resistance of the interior. These values did not change upon fertilization. No spontaneous rhythmical impedance changes such as have been found by Hubbard and Rothschild in the trout egg were found in frog eggs. PMID:19873312
Samgina, Tatyana Yu; Gorshkov, Vladimir A; Artemenko, Konstantin A; Vorontsov, Egor A; Klykov, Oleg V; Ogourtsov, Sergey V; Zubarev, Roman A; Lebedev, Albert T
2012-04-01
Identification of species constituting Rana esculenta complex represents a certain problem as two parental species Rana ridibunda and Rana lessonae form their hybrid R. esculenta, while external signs and sizes of the members of this complex are intersected. However the composition of skin secretion consisting mainly of peptides is different for the species of the complex. LC-MS/MS is an ideal analytical tool for the quantitative and qualitative analysis of these peptides. The results covering elemental composition of these peptides, their levels in the secretion, as well as their belonging to a certain family of peptides may be visualized by means of 2D mass maps. The proposed approach proved itself to be a perspective tool for the reliable identification of all 3 species constituting R. esculenta complex. Easy distinguishing between the species may be achieved using 2D maps as fingerprints. Besides this approach may be used to study hybridogenesis and mechanisms of hemiclonal transfer of genetic information, when rapid and reliable identification of species involved in the process is required. Copyright © 2012 Elsevier Inc. All rights reserved.
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. © 2015 The Authors.
Development of tectal connectivity across metamorphosis in the bullfrog (Rana catesbeiana).
Horowitz, Seth S; Simmons, Andrea Megela
2010-01-01
In the bullfrog (Rana catesbeiana), the process of metamorphosis culminates in the appearance of new visual and visuomotor behaviors reflective of the emergence of binocular vision and visually-guided prey capture behaviors as the animal transitions to life on land. Using several different neuroanatomical tracers, we examined the substrates that may underlie these behavioral changes by tracing the afferent and efferent connectivity of the midbrain optic tectum across metamorphic development. Intratectal, tectotoral, tectotegmental, tectobulbar, and tecto-thalamic tracts exhibit similar trajectories of neurobiotin fiber label across the developmental span from early larval tadpoles to adults. Developmental variability was apparent primarily in intensity and distribution of cell and puncta label in target nuclei. Combined injections of cholera toxin subunit β and Phaseolus vulgaris leucoagglutinin consistently label cell bodies, puncta, or fiber segments bilaterally in midbrain targets including the pretectal gray, laminar nucleus of the torus semicircularis, and the nucleus of the medial longitudinal fasciculus. Developmentally stable label was observed bilaterally in medullary targets including the medial vestibular nucleus, lateral vestibular nucleus, and reticular gray, and in forebrain targets including the posterior and ventromedial nuclei of the thalamus. The nucleus isthmi, cerebellum, lateral line nuclei, medial septum, ventral striatum, and medial pallium show more developmentally variable patterns of connectivity. Our results suggest that even during larval development, the optic tectum contains substrates for integration of visual with auditory, vestibular, and somatosensory cues, as well as for guidance of motivated behaviors. Copyright © 2011 S. Karger AG, Basel.
Toxic effects of endrin and toxaphene on the southern leopard frog Rana sphenocephala
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.
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
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.
First report of Hepatozoon sp. in the Oregon spotted frog, Rana pretiosa.
Stenberg, Patricia L; Bowerman, William J
2010-07-01
From 2005 through 2008, we screened 650 Oregon spotted frogs (Rana pretiosa) from three populations in central Oregon, USA, for hemoparasites. A Hepatozoon sp., not previously reported in R. pretiosa, was found in one population of frogs, mostly as intracellular gamonts at a prevalence of 10.5% with parasitemias ranging from 0.02% to greater than 42% of erythrocytes within individual frogs. Intra-and extracellular merozoites were present, but rare. A potential vector, the mosquito Culex boharti, was common throughout the habitat of the population carrying Hepatozoon sp.
Bkhache, Meriem; Tmimi, Fatim-Zohra; Charafeddine, Omar; Benabdelkrim Filali, Oumama; Lemrani, Meryem; Labbé, Pierrick; Sarih, M'hammed
2018-06-09
Arboviruses are controlled through insecticide control of their mosquito vector. However, inconsiderate use of insecticides often results in the selection of resistance in treated populations, so that monitoring is required to optimize their usage. Here, Culex pipiens (West Nile and Rift Valley Fever virus vector) specimens were collected from four Moroccan cities. Levels of susceptibility to the organophosphate (OP) insecticide malathion were assessed using WHO-recommended bioassays. Individual mosquitoes were tested for the presence of the G119S mutation in the ace-1 gene, the main OP-target resistance mutation. Bioassays showed that mosquitoes from Mohammedia were significantly more resistant to malathion than those from Marrakech. Analyzing the ace-1 genotypes in dead and surviving individuals suggested that other resistance mechanisms may be present in Mohammedia. The ace-1 resistance allele frequencies were relatively moderate (<0.4). Their analyses in three Moroccan cities (Tangier, Casablanca and Marrakech) however showed disparities between two coexisting Cx. pipiens forms and revealed that the G119S mutation tends to be more frequent in urban than in rural collections sites. These findings provide a reference assessment of OP resistance in Morocco and should help the health authorities to develop informed and sustainable vector control programs. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.
Interdigitating cells in the thymus of the frog, Rana temporaria.
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.
Morningstar, Rebecca J; Hamer, Gabriel L; Goldberg, Tony L; Huang, Shaoming; Andreadis, Theodore G; Walker, Edward D
2012-05-01
Analysis of molecular genetic diversity in nine marker regions of five genes within the bacteriophage WO genomic region revealed high diversity of the Wolbachia pipentis strain wPip in a population of Culex pipiens L. sampled in metropolitan Chicago, IL. From 166 blood fed females, 50 distinct genetic profiles of wPip were identified. Rarefaction analysis suggested a maximum of 110 profiles out of a possible 512 predicted by combinations of the nine markers. A rank-abundance curve showed that few strains were common and most were rare. Multiple regression showed that markers associated with gene Gp2d, encoding a partial putative capsid protein, were significantly associated with ancestry of individuals either to form molestus or form pipiens, as determined by prior microsatellite allele frequency analysis. None of the other eight markers was associated with ancestry to either form, nor to ancestry to Cx. quinquefasciatus Say. Logistic regression of host choice (mammal vs. avian) as determined by bloodmeal analysis revealed that significantly fewer individuals that had fed on mammals had the Gp9a genetic marker (58.5%) compared with avian-fed individuals (88.1%). These data suggest that certain wPip molecular genetic types are associated with genetic admixturing in the Cx. pipiens complex of metropolitan Chicago, IL, and that the association extends to phenotypic variation related to host preference.
Mosquitoes established in Lhasa city, Tibet, China
2013-01-01
Background In 2009, residents of Lhasa city, Tibet Autonomous Region (TAR), China reported large numbers of mosquitoes and bites from these insects. It is unclear whether this was a new phenomenon, which species were involved, and whether these mosquitoes had established themselves in the local circumstances. Methods The present study was undertaken in six urban sites of Chengguan district Lhasa city, Tibet. Adult mosquitoes were collected by bed net trap, labor hour method and light trap in August 2009 and August 2012. The trapped adult mosquitoes were initially counted and identified according to morphological criteria, and a proportion of mosquitoes were examined more closely using a multiplex PCR assay. Results 907 mosquitoes of the Culex pipiens complex were collected in this study. Among them, 595 were females and 312 were males. There was no significant difference in mosquito density monitored by bed net trap and labor hour method in 2009 and 2012. Of 105 mosquitoes identified by multiplex PCR, 36 were pure mosquitoes (34.29%) while 69 were hybrids (65.71%). The same subspecies of Culex pipiens complex were observed by bed net trap, labor hour method and light trap in 2009 and 2012. Conclusion The local Culex pipiens complex comprises the subspecies Cx. pipiens pipiens, Cx. pipiens pallens, Cx. pipiens quinquefasciatus and its hybrids. Mosquitoes in the Cx. pipiens complex, known to be, potentially, vectors of periodic filariasis and encephalitis, are now present from one season to the next, and appear to be established in Lhasa City, TAR. PMID:24060238
Effects of photoinduced toxicity of fluoranthene on amphibian embryos and larvae
DOE Office of Scientific and Technical Information (OSTI.GOV)
Hatch, A.C.; Burton, G.A. Jr.
1998-09-01
Embryos and newly hatched larvae of three amphibian species, the spotted salamander (Ambystoma maculatum), the northern leopard frog (Rana pipiens), and the African clawed frog (Xenopus laevis), were exposed to fluoranthene and ultraviolet (UV) light in two scenarios. Embryos were exposed in a laboratory setting from an early developmental stage through hatching under artificial UV light, and newly hatched larvae were exposed outdoors in varying sunlight intensity levels. Outdoor exposures indicated greater sensitivity in the toxic response than did laboratory exposures. In the laboratory, mortality and malformation of X. laevis were the most sensitive indicators of exposure. Xenopus laevis wasmore » also the most sensitive species tested to the effects of UV light alone. Hatching success of R. pipiens was monitored outdoors and was not a useful predictive endpoint in the determination of photoinduced toxicity; however, newly hatched larvae were sensitive to the effects of photoinduced toxicity. Amybstoma maculatum and X. laevis larvae were affected by low ({micro}g/L) concentrations of fluoranthene in sunlight. These findings suggest that low levels of polycyclic aromatic hydrocarbons could be acting synergistically with environmental factors such as UV light to place young amphibians at risk.« less
Neural correlates of frog calling: production by two semi-independent generators.
Schmidt, R S
1992-09-28
The anterior preoptic nuclei of the isolated brainstem of male, Northern leopard frogs (Rana p. pipiens) were stimulated electrically and neural correlates of mating calling recorded from the rhombencephalic mating calling pattern generator. Lesions of discrete areas of the brainstem showed that the mating calling generator is separable into two generators, the pretrigeminal nucleus and the classical pulmonary respiration generator (which is approximately co-extensive with the motor nuclei IX-X). Each of these still can produce pulses when isolated from the other. Their interaction changes the expiratory phase of breathing into the vocal phase of calling. All stages of intermediates between these phases could be seen. An updated and simplified model of call production and evolution is presented.
Stephen C. Richter; Jeanne E. Young; Richard A. Seigal; Glen N. Johnson
2001-01-01
Conservation plans for amphibians often focus on activities at the breeding site, but for species that use temstrial habitats for much of the year, an understanding of nonbreeding habitat use is also essential. We used radio telemetry to study the postbreeding movements of individuals of the only known population of dark gopher frogs, Rana sevosa,...
Cadmium pollution and amphibians--Studies in tadpoles of Rana limnocharis.
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. Copyright © 2015 Elsevier Ltd. All rights reserved.
Pond and landscape determinants of Rana dalmatina population sizes in a Romanian rural landscape
NASA Astrophysics Data System (ADS)
Hartel, Tibor; Nemes, Szilárd; Cogălniceanu, Dan; Öllerer, Kinga; Moga, Cosmin Ioan; Lesbarrères, David; Demeter, László
2009-01-01
Amphibians are good indicators of human impact, declining steadily worldwide. We explored the relationships between the ponds and nearby landscape parameters and population size of the Agile Frog ( Rana dalmatina), estimated from the number of egg masses, in a cultural landscape within the central section of Târnava Mare Basin, Romania. Forty-three permanent ponds were surveyed in a 2600 km 2 area. The average number of egg masses per pond was 211.13 (SD = 426.41). The egg mass number was significantly and positively related to the emergent aquatic macrophyte cover (its effect peaks at around 50%) and the green connecting corridors between the ponds and forests, and negatively related to the extent of nearby urban areas. The proximity of the forest (positive effect) and the presence of high traffic roads (negative effect) were highly correlated with green corridors and further eliminated from the model due to multicollinearity. Both these variables had significant effects when incorporated in univariate models and multivariate models without green corridors. Since a large part of our study area was currently declared as Natura 2000 site, there is an increased need for management proposals and conservation applications for biodiversity, including amphibians. Rana dalmatina is an important species for monitoring because it is common in the studied area and is suited for short surveys.
Brustolin, M; Talavera, S; Santamaría, C; Rivas, R; Pujol, N; Aranda, C; Marquès, E; Valle, M; Verdún, M; Pagès, N; Busquets, N
2016-06-01
The emerging disease West Nile fever is caused by West Nile virus (WNV), one of the most widespread arboviruses. This study represents the first test of the vectorial competence of European Culex pipiens Linnaeus 1758 and Stegomyia albopicta (= Aedes albopictus) (both: Diptera: Culicidae) populations for lineage 1 and 2 WNV isolated in Europe. Culex pipiens and S. albopicta populations were susceptible to WNV infection, had disseminated infection, and were capable of transmitting both WNV lineages. This is the first WNV competence assay to maintain mosquito specimens under environmental conditions mimicking the field (day/night) conditions associated with the period of maximum expected WNV activity. The importance of environmental conditions is discussed and the issue of how previous experiments conducted in fixed high temperatures may have overestimated WNV vector competence results with respect to natural environmental conditions is analysed. The information presented should be useful to policymakers and public health authorities for establishing effective WNV surveillance and vector control programmes. This would improve preparedness to prevent future outbreaks. © 2016 The Authors. Medical and Veterinary Entomology published by John Wiley & Sons Ltd on behalf of Royal Entomological Society.
Elizabeth A. Roznik; Steve A. Johnson; Cathryn H. Greenberg; George W. Tanner
2009-01-01
Many animals exhibit changes in patterns of movement and habitat use as they age, and understanding such ontogenetic shifts is important for ensuring that habitat management is appropriate for all life stages. We used radiotelemetry to study movements and habitat use of juvenile and adult gopher frogs (Rana capito) as they migrated from the same ponds following...
S. Kupferberg; A. Lind; V. Thill; S. Yarnell
2011-01-01
We explored the effects of large magnitude flow fluctuations in rivers with dams, commonly referred to as pulsed flows, on tadpoles of the lotic-breeding Foothill Yellow-legged Frog, Rana boylii. We quantified the velocity conditions in habitats occupied by tadpoles and then conducted experiments to assess the tolerance to values at the upper limit...
Daniel Saenz; James B. Johnson; Cory K. Adams; Gage H. Dayton
2003-01-01
Phenotypic plasticity, such as morphological and behavioral changes in response to predators, is common in larval anurans. Less is known about inducible defenses in the embryonic stages of development. We investigated the predation risk imposed by crayfish (Procambarus nigrocinctus) on southern leopard frog (Rana sphenocephala)...
K.L. Pope; K.R. Matthews
2001-01-01
Movement ecology and seasonal distribution of mountain yellow-legged frogs (Rana muscosa) in Dusy Basin (3470 m), Kings Canyon National Park, California, were characterized using passive integrated transponder (PIT) surveys and visual encounter surveys. We individually PIT-tagged 500 frogs during the summers of 1997 and 1998 and monitored these individuals during seven...
A.J. Lind; H.B. Shaffer; P.Q. Spinks; G.M. Fellers
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...
Terra Fuller; Karen Pope; Donald Ashton; Hartwell Welsh
2010-01-01
Extensive modifications of river systems have left floodplains some of the most endangered ecosystems in the world and made restoration of these systems a priority. Modified river ecosystems frequently support invasive species to the detriment of native species. Rana catesbeiana (American bullfrog) is an invasive amphibian that thrives in modified...
Liver lesions produced by aflatoxins in Rana catesbeiana (bullfrog).
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.
Tao, Li-Ming; Yang, Jian-Zhong; Zhuang, Pei-Jun; Tang, Zhen-Hua
2006-01-01
A malathion-resistant (RM) strain of Culex pipiens pallens Coq was obtained by successively selecting a field population with malathion in the laboratory. The synergistic effect of iprobenfos on malathion toxicity and alpha-naphthyl acetate (alpha-NA) esterase assay revealed that malathion resistance in the RM strain was associated with increased alpha-NA esterase activity and the synergism was mainly due to the inhibition by iprobenfos of this activity. There was no difference in alpha-NA esterase activity between the larvae and female adults in the susceptible (S) strain, but the activity in the adults was 13-fold higher than in the larvae of the RM strain. To understand the effect of the application of a mixture of iprobenfos and malathion on the evolution of malathion resistance, an artificial strain (Syn) was generated by mixing the RM and S strains with 0.1 frequency of the malathion-resistant individuals. The offspring of the Syn strain were divided into two sub-strains, Rm and Rm+ibp, which were successively treated with, respectively, malathion alone and malathion + iprobenfos (1:2) at LC70. In the mixture, the fungicide iprobenfos acted as a synergist of malathion. After treatment for 10 generations, the resistance level to malathion was 317.4-fold for the Rm sub-strain, whereas for the Rm+ibp sub-strain it was only 38.9-fold, compared with the Syn strain. Similar results were obtained by measurement of alpha-NA esterase activity from both larvae and female adults. The alpha-NA esterase activities in larvae and female adults at F10 generation were 2.6- and 10.9-fold from the Rm+ibp sub-strain and 5.7- and 98.5-fold from the Rm sub-strain, respectively, compared with the Syn strain. The above results suggested that iprobenfos, although it cannot completely stop or prevent the onset of malathion resistance, could dramatically delay its evolution. Copyright 2005 Society of Chemical Industry.
Citrus essential oils and four enantiomeric pinenes against Culex pipiens (Diptera: Culicidae).
Michaelakis, Antonios; Papachristos, Dimitrios; Kimbaris, Athanasios; Koliopoulos, George; Giatropoulos, Athanasios; Polissiou, Moschos G
2009-09-01
The aim of this study was to evaluate the toxicity of pinenes (enantiomers of alpha- and beta-) and essential oils from Greek plants of the Rutaceae family against the mosquito larvae of Culex pipiens (Diptera: Culicidae). Essential oils were isolated by hydrodistillation from fruit peel of orange (Citrus sinensis L.), lemon (Citrus limon L.), and bitter orange (Citrus aurantium L.). The chemical composition was determined by gas chromatography/mass spectrometry (GC/MS) analysis. Citrus essential oils contained in high proportion limonene and in lower quantities p-menthane molecules and pinenes. The insecticidal action of these essential oils and enantiomers of their pinenes on mosquito larvae was evaluated. Plant essential oils exhibited strong toxicity against larvae with the LC(50) values ranging from 30.1 (lemon) to 51.5 mg/L (orange) depending on Citrus species and their composition. Finally, the LC(50) value of pinenes ranging from 36.53 to 66.52 mg/L indicated an enantioselective toxicity only for the beta-pinene enantiomer.
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.
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.
Gary M. Fellers; Karen L. Pope; Jonathan E. Stead; Michelle S. Koo; Hartwell H. Welsh
2007-01-01
Monitoring the distribution, population size, and trends of declining species is necessary to evaluate their vulnerability to extinction. It is the responsibility of scientists to alert management professionals of the need for preemptive action if a species approaches imminent, regional extirpation. This is the case with Rana cascadae (Cascades Frog...
Population trends, survival, and sampling methodologies for a population of Rana draytonii
Fellers, Gary M.; Kleeman, Patrick M.; Miller, David A.W.; Halstead, Brian J.
2017-01-01
Estimating population trends provides valuable information for resource managers, but monitoring programs face trade-offs between the quality and quantity of information gained and the number of sites surveyed. We compared the effectiveness of monitoring techniques for estimating population trends of Rana draytonii (California Red-legged Frog) at Point Reyes National Seashore, California, USA, over a 13-yr period. Our primary goals were to: 1) estimate trends for a focal pond at Point Reyes National Seashore, and 2) evaluate whether egg mass counts could reliably estimate an index of abundance relative to more-intensive capture–mark–recapture methods. Capture–mark–recapture (CMR) surveys of males indicated a stable population from 2005 to 2009, despite low annual apparent survival (26.3%). Egg mass counts from 2000 to 2012 indicated that despite some large fluctuations, the breeding female population was generally stable or increasing, with annual abundance varying between 26 and 130 individuals. Minor modifications to egg mass counts, such as marking egg masses, can allow estimation of egg mass detection probabilities necessary to convert counts to abundance estimates, even when closure of egg mass abundance cannot be assumed within a breeding season. High egg mass detection probabilities (mean per-survey detection probability = 0.98 [0.89–0.99]) indicate that egg mass surveys can be an efficient and reliable method for monitoring population trends of federally threatened R. draytonii. Combining egg mass surveys to estimate trends at many sites with CMR methods to evaluate factors affecting adult survival at focal populations is likely a profitable path forward to enhance understanding and conservation of R. draytonii.
Graydon, Cole W; Cho, Soyoun; Diamond, Jeffrey S; Kachar, Bechara; von Gersdorff, Henrique; Grimes, William N
2014-06-11
Sensory processing in the auditory system requires that synapses, neurons, and circuits encode information with particularly high temporal and spectral precision. In the amphibian papillia, sound frequencies up to 1 kHz are encoded along a tonotopic array of hair cells and transmitted to afferent fibers via fast, repetitive synaptic transmission, thereby promoting phase locking between the presynaptic and postsynaptic cells. Here, we have combined serial section electron microscopy, paired electrophysiological recordings, and Monte Carlo diffusion simulations to examine novel mechanisms that facilitate fast synaptic transmission in the inner ear of frogs (Rana catesbeiana and Rana pipiens). Three-dimensional anatomical reconstructions reveal specialized spine-like contacts between individual afferent fibers and hair cells that are surrounded by large, open regions of extracellular space. Morphologically realistic diffusion simulations suggest that these local enlargements in extracellular space speed transmitter clearance and reduce spillover between neighboring synapses, thereby minimizing postsynaptic receptor desensitization and improving sensitivity during prolonged signal transmission. Additionally, evoked EPSCs in afferent fibers are unaffected by glutamate transporter blockade, suggesting that transmitter diffusion and dilution, and not uptake, play a primary role in speeding neurotransmission and ensuring fidelity at these synapses. Copyright © 2014 the authors 0270-6474/14/348358-15$15.00/0.
James B. Johnson; Daniel Saenz; Cory K. Adams; Richard N. Conner
2003-01-01
Abstract: We tested the hypotheses that potential egg predators, crayfish Procambarus nigrocinctus and dytiscid Cybister sp. larvae, would accelerate the timing of hatching and that a larval predator, dragonfly naiad Anax junius, would delay hatching in the southern leopard frog (Rana...
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...
Hyperforin and deoxycohumulone as a larvicidal agent against Culex pipiens (Diptera: Culicidae).
Mitsopoulou, Kornilia P; Vidali, Veroniki P; Koliopoulos, George; Couladouros, Elias A; Michaelakis, Antonios
2014-04-01
The larvicidal effect of hyperforin (1), a bioactive compound of Hypericum perforatum, and deoxycohumulone (2) (biosynthetic precursor of hyperforin) were evaluated against Culex pipiens (Diptera: Culicidae) for the first time. All the acetate analogues (3-6) of hyperforin (1) and deoxycohumulone (2) were also synthesized and bioassayed to provide information on structural requirements for the tested compounds. Larvicidal results revealed that hyperforin (1) and deoxycohumulone (2) exhibited potent activity with LC50 value of 26.72 and 51.03 mg L(-1), respectively. The monoacetyl-deoxycohumulone (4) displayed lower activity with LC50 value of 135.92 mg L(-1), while all other acetate analogues were inactive at concentrations even as high as 150 mg L(-1), indicating that the free hydroxyl groups are essential for the larvicidal activity. The mortality values were increased, more than 80%, when 10 mg L(-1) piperonyl butoxide were added in hyperforin (1) or deoxycohumulone (2) bioassays. Finally, sub-lethal survival analysis is conducted for three doses of hyperforin (1) and deoxycohumulone (2) and results are discussed. Copyright © 2013 Elsevier Ltd. All rights reserved.
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.
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
Habitat use and home range of the endangered gold-spotted pond frog (Rana chosenica).
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.
Traboulsi, Abdallah F; Taoubi, K; el-Haj, Samih; Bessiere, J M; Rammal, Salma
2002-05-01
The insecticidal activities of essential oil extracts from leaves and flowers of aromatic plants against fourth-instar larvae of the mosquito Culex pipiens molestus Forskal were determined. Extracts of Myrtus communis L were found to be the most toxic, followed by those of Origanum syriacum L, Mentha microcorphylla Koch, Pistacia lentiscus L and Lavandula stoechas L with LC50 values of 16, 36, 39, 70 and 89 mg litre-1, respectively. Over 20 major components were identified in extracts from each plant species. Eight pure components (1,8-cineole, menthone, linalool, terpineol, carvacrol, thymol, (1S)-(-)-alpha-pinene and (1R)-(+)-alpha-pinene) were tested against the larvae. Thymol, carvacrol, (1R)-(+)-alpha-pinene and (1S)-(-)-alpha-pinene were the most toxic (LC50 = 36-49 mg litre-1), while menthone, 1,8-cineole, linalool and terpineol (LC50 = 156-194 mg litre-1) were less toxic.
Wassersug, R J; Feder, M E
1983-07-01
Larvae of the anurans Rana berlandieri and Xenopus laevis have lungs and can breathe air as well as irrigate buccal and pharyngeal surfaces for aquatic respiration. Larvae of Bufo americanus lack lungs until just before metamorphosis and are obligately aquatic. We examined the relationship between the locomotor stamina (time to fatigue), aquatic oxygen concentration, body size, and respiratory behaviour of swimming larvae of these species, with the following results: Stamina is size-dependent in all three species. Aquatic hypoxia reduces stamina in larvae of all three species, but most conspicuously in Bufo. Breathing air increases stamina in Rana larvae, especially in large animals and under aquatic hypoxia. In contrast to Rana larvae, Xenopus larvae swimming in normoxic water undergo a reduction in stamina when allowed to breathe air. In hypoxic water, aerial respiration moderates the reduction in stamina seen in Xenopus larvae. Branchial irrigation is associated with increased stamina in Xenopus, and is increased under hypoxia and at high swimming velocities. Respiratory demand, buoyancy and the drag associated with branchial irrigation all affect respiratory behaviour in Xenopus larvae. The great amount of interspecific variation in the relationship between respiratory behaviour and stamina reveals the importance of measuring performance directly when attempting to interpret the functional significance of respiratory structures and behaviour.
Incorporation of Tritium-labelled Thymidine in Bufo $female$ × Rana temporaria $male$ Hybrid Embryos
DOE Office of Scientific and Technical Information (OSTI.GOV)
TENCER, B.
1961-04-01
Two-cell stages of hybrid embryos resulting from the cross-fertilization of Bufo and Rana temporaria were incubated for 17 hrs in a medium containing tritium-labeled thymidine. The embryos were fixed by freeze-substitution and the incorporation of tritium studied by the radioautographic technique. The embryos stopped development at the late blastula stage. Labeling of desoxyribonucleic acid was demonstrated in morula as well as in blastula cells of the lethal hybrids. Tritium-labeled thymidine was shown to be incorporated into desoxyribonucleic acid 24 hr after development stopped, which suggests that the block in development was not due to the arrest of desoxyribonucleic acid synthesis.more » (C.H.)« less
Landscape associations of frog and toad species in Iowa and Wisconsin, U.S.A
Knutson, M.G.; Sauer, J.R.; Olsen, D.A.; Mossman, M.J.; Hemesath, L.M.; Lannoo, M.J.; Kaiser, Hinrich; Casper, Gary S.; Bernstein, Neil P.
2000-01-01
Landscape habitat associations of frogs and toads in Iowa and Wisconsin were tested to determine whether they support or refute previous general habitat classifications. We examined which Midwestern species shared similar habitats to see if these associations were consistent across large geographic areas (states). Rana sylvatica (wood frog), Hyla versicolor (eastern gray treefrog), Pseudacris crucifer (spring peeper), and Acris crepitans (cricket frog) were identified as forest species, P. triseriata (chorus frog), H. chrysoscelis (Cope's gray treefrog), R. pipiens (leopard frog), and Bufo americanus (American toad) as grassland species, and R. catesbeiana (bullfrog), R. clamitans (green frog), R. palustris (pickerel frog), and R. septentrionalis (mink frog) as lake or stream species. The best candidates to serve as bioindicators of habitat quality were the forest species R. sylvatica, H. versicolor, and P. crucifer, the grassland species R. pipiens and P. triseriata, and a cold water wetland species, R. palustris. Declines of P. crucifer, R. pipiens, and R. palustris populations in one or both states may reflect changes in habitat quality. Habitat and community associations of some species differed between states, indicating that these relationships may change across the range of a species. Acris crepitans may have shifted its habitat affinities from open habitats, recorded historically, to the more forested habitat associations we recorded. We suggest contaminants deserve more investigation regarding the abrupt and widespread declines of this species. Interspersion of different habitat types was positively associated with several species. A larger number of wetland patches may increase breeding opportunities and increase the probability of at least one site being suitable. We noted consistently negative associations between anuran species and urban development. Given the current trend of urban growth and increasing density of the human population, declines of
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.
Regeneration of the eighth cranial nerve in the bullfrog, Rana catesbeiana.
Newman, A; Honrubia, V
1992-01-01
The present study was done in order to document the ability of the eighth cranial nerve of the bullfrog (Rana catesbeiana) to regenerate, the anatomic characteristics of the regenerated fibers, and the specificity of projections from individual endorgan branches of the nerve. The eighth cranial nerve was sharply transected between the ganglion cells and the brain stem in 40 healthy bullfrogs and allowed to regenerate. Anatomic studies were performed in these animals a minimum of 3 months postoperatively. Horseradish peroxidase was used to label the whole vestibular nerve or its individual endorgan branches. Labeled regenerated fibers could be identified crossing the site of the nerve section and projecting centrally to the vestibular nuclei in a pattern similar to that of normal frogs. Labeling of individual branches showed that regenerated fibers innervated the same specific areas found in normal frogs. Unlike normal animals, both thick and thin fibers projected to the medial nucleus.
Cancrini, G; Scaramozzino, P; Gabrielli, S; Di Paolo, M; Toma, L; Romi, R
2007-11-01
To identify the natural vectors of Dirofilaria repens Railliet et Henry, entomological samplings were carried out in four sites within the Lazio region, foci of canine subcutaneous dirofilariasis. Collections were made in 2002-2003 by means of dog-baited and miniature Centers for Disease Control and Prevention traps as well as on humans. Microscopy identified 1576 attracted mosquito females as belonging to six species, but molecular diagnostics detected filarial DNA only in Culex pipiens L. and Aedes albopictus (Skuse, 1894). Dirofilaria immitis Leidy DNA, D. repens DNA, or both were found in the head and thorax of both mosquitoes. The simultaneous presence of vectors showing diurnal and nocturnal activity patterns is of concern for animal and human health. The finding of D. immitis DNA in mosquitoes in areas where only D. repens was been recovered in dogs also demonstrates that this filarial parasite circulates among carnivores (wild or domesticated pets).
Patterns of protein synthesis in oocytes and early embryos of Rana esculenta complex.
Chen, P S; Stumm-Zollinger, E
1986-01-01
We have used isotopic labelling and both one-and two-dimensional electrophoretic procedures to analyse the protien synthesis patterns in oocytes and early embryos of three phenotypes of the European green frogs. The results demonstrated that protein patterns of Rana ridibunda and R. esculenta are identical, but that they differ from those of R. lessonae. Progeny of the lethal cross R. esculenta × R. esculenta showed a distinct delay in the appearance of stage-specific proteins during early embryogenesis. The heat-shock response of R. ridibunda and R. esculenta oocytes was found to be identical, but different from that of Xenopus laevis. The implications of these findings, with respect to hybridogenesis in R. esculenta complex and variations in the regulations of heat shock genes in different amphibian species, are discussed.
Ahmed, Ashraf M; Hussein, Hamdy I; El-Kersh, Talat A; Al-Sheikh, Yazeed A; Ayaad, Tahany H; El-Sadawy, Hanan A; Al-Mekhlafi, Fahd A; Ibrahim, Mohamed S; Al-Tamimi, Jameel; Nasr, Fahd A
2017-06-01
The incidence of mosquito-borne diseases and the resistance of mosquitoes to conventional pesticides have recently caused a panic to the authorities in the endemic countries. This study was conducted to identify native larvicidal biopesticides against Culex pipiens for utilization in the battle against mosquito-borne diseases. Larvicidal activities of new indigenous Bacillus thuringiensis isolates and crude toxin complexes (TCs) of two nematode bacterial-symbionts, Photorhabdus luminescens akhurstii (HRM1) and Ph. luminescens akhurstii (HS1) that tested against Cx. pipiens . B. thuringiensis isolates were recovered from different environmental samples in Saudi Arabia, and the entomopathogenic nematodes, Heterorhabditis indica (HRM1) and He. sp (HS1) were isolated from Egypt. Larvicidal activities (LC 50 and LC 95 ) of the potentially active B. thuringiensis strains or TCs were then evaluated at 24 and 48h post-treatment. Three B. thuringiensis isolates were almost as active as the reference B. thuringiensis israelensis (Bti-H14), and seven isolates were 1.6-5.4 times more toxic than Bti-H14. On the other hand, the TCs of the bacterial symbionts, HRM1 and HS1, showed promising larvicidal activities. HS1 showed LC 50 of 2.54 folds that of HRM1 at 24h post-treatment. Moreover, histopathological examinations of the HS1-treated larvae showed deformations in midgut epithelial cells at 24h post-treatment. Synergistic activity and molecular characterization of these potentially active biocontrol agents are currently being investigated. These results may lead to the identification of eco-friend mosquito larvicidal product(s) that could contribute to the battle against mosquito-borne diseases.
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.
Ahmed, Ashraf M; Hussein, Hamdy I; El-Kersh, Talat A; Al-Sheikh, Yazeed A; Ayaad, Tahany H; El-Sadawy, Hanan A; Al-Mekhlafi, Fahd A; Ibrahim, Mohamed S; Al-Tamimi, Jameel; Nasr, Fahd A
2017-01-01
Background: The incidence of mosquito-borne diseases and the resistance of mosquitoes to conventional pesticides have recently caused a panic to the authorities in the endemic countries. This study was conducted to identify native larvicidal biopesticides against Culex pipiens for utilization in the battle against mosquito-borne diseases. Methods: Larvicidal activities of new indigenous Bacillus thuringiensis isolates and crude toxin complexes (TCs) of two nematode bacterial-symbionts, Photorhabdus luminescens akhurstii (HRM1) and Ph. luminescens akhurstii (HS1) that tested against Cx. pipiens. B. thuringiensis isolates were recovered from different environmental samples in Saudi Arabia, and the entomopathogenic nematodes, Heterorhabditis indica (HRM1) and He. sp (HS1) were isolated from Egypt. Larvicidal activities (LC50 and LC95) of the potentially active B. thuringiensis strains or TCs were then evaluated at 24 and 48h post-treatment. Results: Three B. thuringiensis isolates were almost as active as the reference B. thuringiensis israelensis (Bti-H14), and seven isolates were 1.6–5.4 times more toxic than Bti-H14. On the other hand, the TCs of the bacterial symbionts, HRM1 and HS1, showed promising larvicidal activities. HS1 showed LC50 of 2.54 folds that of HRM1 at 24h post-treatment. Moreover, histopathological examinations of the HS1-treated larvae showed deformations in midgut epithelial cells at 24h post-treatment. Conclusion: Synergistic activity and molecular characterization of these potentially active biocontrol agents are currently being investigated. These results may lead to the identification of eco-friend mosquito larvicidal product(s) that could contribute to the battle against mosquito-borne diseases. PMID:29062851
Landscape resistance to frog movements
Mazerolle, M.J.; Desrochers, A.
2005-01-01
An animal's capacity to recolonize a patch depends on at least two components: its ability to detect the patch and its ability to reach it. However, the disruption of such processes by anthropic disturbances could explain low animal abundance patterns observed by many investigators in certain landscapes. Through field experiments, we compared the orientation and homing success of northern green frogs (Rana clamitans melanota Rafinesque, 1820) and northern leopard frogs (Rana pipiens Schreber, 1782) translocated across disturbed or undisturbed surfaces. We also monitored the path selected by individuals when presented with a choice between a short distance over a disturbed surface and a longer, undisturbed route. Finally, we measured the water loss and behaviour of frogs on substrates resulting from anthropogenic disturbances and a control. When presented with a choice, 72% of the frogs avoided disturbed surfaces. Although able to orient towards the pond of capture when translocated on disturbed surfaces, frogs had a lower probability of homing successfully to the pond than when translocated at a similar distance on an undisturbed surface. Frogs lost the most water on substrates associated with disturbance and in the absence of cover. Our data illustrate that anthropically disturbed areas devoid of cover, such as mined peatlands and agricultural fields, disrupt the ability of frogs to reach habitat patches and are likely explanations to their reduced abundance patterns in such environments. ?? 2005 NRC Canada.
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.
Muir, Anna P; Biek, Roman; Mable, Barbara K
2014-05-23
Extreme environments can impose strong ecological and evolutionary pressures at a local level. Ectotherms are particularly sensitive to low-temperature environments, which can result in a reduced activity period, slowed physiological processes and increased exposure to sub-zero temperatures. The aim of this study was to assess the behavioural and physiological responses that facilitate survival in low-temperature environments. In particular, we asked: 1) do high-altitude common frog (Rana temporaria) adults extend the time available for larval growth by breeding at lower temperatures than low-altitude individuals?; and 2) do tadpoles sampled from high-altitude sites differ physiologically from those from low-altitude sites, in terms of routine metabolic rate (RMR) and freeze tolerance? Breeding date was assessed as the first day of spawn observation and local temperature recorded for five, paired high- and low-altitude R. temporaria breeding sites in Scotland. Spawn was collected and tadpoles raised in a common laboratory environment, where RMR was measured as oxygen consumed using a closed respiratory tube system. Freeze tolerance was measured as survival following slow cooling to the point when all container water had frozen. We found that breeding did not occur below 5°C at any site and there was no significant relationship between breeding temperature and altitude, leading to a delay in spawning of five days for every 100 m increase in altitude. The relationship between altitude and RMR varied by mountain but was lower for individuals sampled from high- than low-altitude sites within the three mountains with the highest high-altitude sites (≥900 m). In contrast, individuals sampled from low-altitudes survived freezing significantly better than those from high-altitudes, across all mountains. Our results suggest that adults at high-altitude do not show behavioural adaptations in terms of breeding at lower temperatures. However, tadpoles appear to have the
Fouad, Hatem; Hongjie, Li; Yanmei, Ding; Baoting, Yu; El-Shakh, Ahmed; Abbas, Ghulam; Jianchu, Mo
2017-11-01
Culex pipiens pallens are the most common mosquito's vector in Asia. In order to protect the people from diseases, the anti-mosquito population is necessary that uses safe and new bio-pesticides such as bacteria-AgNPs. In our report, we used two kinds of bacteria to synthesize silver nanoparticles to examine the toxic effect on the larvae and pupae of Cx. pipiens pallens and also used as antimicrobial activity. The biosynthesis of AgNPs and its characterization was carried out by UV-Vis spectrophotometry, FTIR, TEM, SEM, and EDX. The larvicidal and pupicidal assays revealed that the lethal concentration LC 50 values of Bacillus amyloliquefaciens-AgNPs were 0.72 ppm (I), 0.73 ppm (II), 0.69 ppm (III), 1.16 ppm (IV), and 4.18 (Pupae), while LC 50 values of Bacillus subtilis-AgNPs were 0.60 ppm (I), 0.62 ppm (II), 0.21 ppm (III), 0.28 ppm (IV), and 3.46 ppm (Pupae) after 72 h of exposure. Antibacterial activity test of AgNPs reveals better results against rice pathogenic bacteria than bacteria alone. Thus, bacteria-mediated silver nanoparticles have a rapid effect on vector mosquito and microbial pathogen suggesting savings of energy and resources. Hence, bacteria-AgNPs may be used in the future as an effective weapon to control vector mosquito and harmful bacteria.
I Lacan; Kathleen R. Matthews; K.V. Feldman
2008-01-01
Between-year variation in snowpack (from 20 to 200% of average) and summer rainfall cause large fluctuations in volume of small lakes in the higher elevation (> 3000 m) Sierra Nevada, which are important habitat for the imperiled Sierra Nevada Yellow-legged Frog, Rana sierrae. Climate change (global warming) is predicted to increase these...
Panova, Ina G; Sharova, Natalia P; Dmitrieva, Svetlana B; Levin, Peter P; Tatikolov, Alexander S
2008-12-01
This study aimed to analyze the aqueous humor (AH) and the vitreous body (VB) of the eye of the adult frog Rana temporaria L. as a representative species of amphibians, which lead a semi-terrestrial life. The presence of collagen, albumin, uric acid and electron donors was shown in both media; however, there are slight differences in their concentrations. To determine collagen, a spectral-fluorescent probe, cyanine dye, was used. The presence of collagen in AH of the frog was found at the first time. The total content of electron donors (ascorbic and uric acids, tryptophan, and tyrosine) in VB and HA was roughly estimated at approximately 1.5x10(-4) mol/L. Both VB and AH absorb light in similar UV regions. The total protein and albumin contents in AH were found to be somewhat higher than those in VB. The uric acid content was at an equally low level in both intraocular media. It is supposed that the similarity of VB and AH compositions shown in this work is due to some exchange between VB and AH contents in the course of accommodation. The role of intraocular fluids in physiological functions of the eye and in protecting the retina against UV light is discussed.
Rosà, Roberto; Marini, Giovanni; Bolzoni, Luca; Neteler, Markus; Metz, Markus; Delucchi, Luca; Chadwick, Elizabeth A; Balbo, Luca; Mosca, Andrea; Giacobini, Mario; Bertolotti, Luigi; Rizzoli, Annapaola
2014-06-12
West Nile Virus (WNV) is an emerging global health threat. Transmission risk is strongly related to the abundance of mosquito vectors, typically Culex pipiens in Europe. Early-warning predictors of mosquito population dynamics would therefore help guide entomological surveillance and thereby facilitate early warnings of transmission risk. We analysed an 11-year time series (2001 to 2011) of Cx. pipiens mosquito captures from the Piedmont region of north-western Italy to determine the principal drivers of mosquito population dynamics. Linear mixed models were implemented to examine the relationship between Cx. pipiens population dynamics and environmental predictors including temperature, precipitation, Normalized Difference Water Index (NDWI) and the proximity of mosquito traps to urban areas and rice fields. Warm temperatures early in the year were associated with an earlier start to the mosquito season and increased season length, and later in the year, with decreased abundance. Early precipitation delayed the start and shortened the length of the mosquito season, but increased total abundance. Conversely, precipitation later in the year was associated with a longer season. Finally, higher NDWI early in the year was associated with an earlier start to the season and increased season length, but was not associated with abundance. Proximity to rice fields predicted higher total abundance when included in some models, but was not a significant predictor of phenology. Proximity to urban areas was not a significant predictor in any of our models. Predicted variations in start of the season and season length ranged from one to three weeks, across the measured range of variables. Predicted mosquito abundance was highly variable, with numbers in excess of 1000 per trap per year when late season temperatures were low (average 21°C) to only 150 when late season temperatures were high (average 30°C). Climate data collected early in the year, in conjunction with local land
Occurrence of avian Plasmodium and West Nile virus in culex species in Wisconsin
Hughes, T.; Irwin, P.; Hofmeister, E.; Paskewitz, S.M.
2010-01-01
The occurrence of multiple pathogens in mosquitoes and birds could affect the dynamics of disease transmission. We collected adult Culex pipiens and Cx. restuans (Cx. pipiens/restuans hereafter) from sites in Wisconsin and tested them for West Nile virus (WNV) and for avian malaria (Plasmodium). Gravid Cx. pipiens/restuans were tested for WNV using a commercial immunoassay, the RAMP?? WNV test, and positive results were verified by reverse transcriptasepolymerase chain reaction. There were 2 WNV-positive pools of Cx. pipiens/restuans in 2006 and 1 in 2007. Using a bias-corrected maximum likelihood estimation, the WNV infection rate for Cx. pipiens/restuans was 5.48/1,000 mosquitoes in 2006 and 1.08/1,000 mosquitoes in 2007. Gravid Cx. pipiens or Cx. restuans were tested individually for avian Plasmodium by a restriction enzymebased assay. Twelve mosquitoes were positive for avian Plasmodium (10.0), 2 were positive for Haemoproteus, and 3 were positive for Leucocytozoon. There were 4 mixed infections, with mosquitoes positive for >1 of the hemosporidian parasites. This work documents a high rate of hemosporidian infection in Culex spp. and illustrates the potential for co-infections with other arboviruses in bird-feeding mosquitoes and their avian hosts. In addition, hemosporidian infection rates may be a useful tool for investigating the ecological dynamics of Culex/avian interactions. ?? 2010 by The American Mosquito Control Association, Inc.
Optical and tomographic imaging of a middle ear malformation in the bullfrog (Rana catesbeiana).
Horowitz, Seth S; Simmons, Andrea Megela; Ketten, Darlene R
2005-08-01
Using a combination of in vivo computerized tomography and histological staining, a middle ear anomaly in two wild-caught American bullfrogs (Rana catesbeiana) is characterized. In these animals, the tympanic membrane, extrastapes, and pars media (shaft) of the stapes are absent on one side of the head, with the other side exhibiting normal morphology. The pars interna (footplate) of the stapes and the operculum are present in their normal positions at the entrance of the otic capsule on both the affected and unaffected sides. The pattern of deformity suggests a partial failure of development of tympanic pathway tissues, but with a preservation of the opercularis pathway. While a definitive proximate cause of the condition could not be determined, the anomalies show similarities to developmental defects in mammalian middle ear formation.
Optical and tomographic imaging of a middle ear malformation in the bullfrog (Rana catesbeiana)
Horowitz, Seth S.; Simmons, Andrea Megela; Ketten, Darlene R.
2005-01-01
Using a combination of in vivo computerized tomography and histological staining, a middle ear anomaly in two wild-caught American bullfrogs (Rana catesbeiana) is characterized. In these animals, the tympanic membrane, extrastapes, and pars media (shaft) of the stapes are absent on one side of the head, with the other side exhibiting normal morphology. The pars interna(footplate) of the stapes and the operculum are present in their normal positions at the entrance of the otic capsule on both the affected and unaffected sides. The pattern of deformity suggests a partial failure of development of tympanic pathway tissues, but with a preservation of theopercularis pathway. While a definitive proximate cause of the condition could not be determined, the anomalies show similarities to developmental defects in mammalian middle ear formation. PMID:16158670
Atyame, Célestine M.; Pasteur, Nicole; Dumas, Emilie; Tortosa, Pablo; Tantely, Michaël Luciano; Pocquet, Nicolas; Licciardi, Séverine; Bheecarry, Ambicadutt; Zumbo, Betty; Weill, Mylène; Duron, Olivier
2011-01-01
The use of the bacterium Wolbachia is an attractive alternative method to control vector populations. In mosquitoes, as in members of the Culex pipiens complex, Wolbachia induces a form of embryonic lethality called cytoplasmic incompatibility, a sperm-egg incompatibility occurring when infected males mate either with uninfected females or with females infected with incompatible Wolbachia strain(s). Here we explore the feasibility of the Incompatible Insect Technique (IIT), a species-specific control approach in which field females are sterilized by inundative releases of incompatible males. We show that the Wolbachia wPip(Is) strain, naturally infecting Cx. p. pipiens mosquitoes from Turkey, is a good candidate to control Cx. p. quinquefasciatus populations on four islands of the south-western Indian Ocean (La Réunion, Mauritius, Grande Glorieuse and Mayotte). The wPip(Is) strain was introduced into the nuclear background of Cx. p. quinquefasciatus mosquitoes from La Réunion, leading to the LR[wPip(Is)] line. Total embryonic lethality was observed in crosses between LR[wPip(Is)] males and all tested field females from the four islands. Interestingly, most crosses involving LR[wPip(Is)] females and field males were also incompatible, which is expected to reduce the impact of any accidental release of LR[wPip(Is)] females. Cage experiments demonstrate that LR[wPip(Is)] males are equally competitive with La Réunion males resulting in demographic crash when LR[wPip(Is)] males were introduced into La Réunion laboratory cages. These results, together with the geographic isolation of the four south-western Indian Ocean islands and their limited land area, support the feasibility of an IIT program using LR[wPip(Is)] males and stimulate the implementation of field tests for a Cx. p. quinquefasciatus control strategy on these islands. PMID:22206033
Relationships between leukocytes and Hepatozoon spp. In green frogs, Rana clamitans.
Shutler, Dave; Smith, Todd G; Robinson, Stephen R
2009-01-01
There are few published data on amphibian leukocyte profiles, and relationships between amphibian leukocytes and parasites are even less well known. Using counts from 35 pairs of blood smears taken 2 days apart, we tested for correlations between leukocyte proportions and infection intensities of Hepatozoon spp. (either Hepatozoon catesbianae or Hepatozoon clamatae) in green frogs (Rana clamitans). On average (SE), we counted 65.4 (1.7) lymphocytes, 14.0 (1.3) neutrophils, 19.3 (1.6) eosinophils, 0.9 (0.1) monocytes, and 0.4 (0.1) basophils per 100 leukocytes. All frogs harbored Hepatozoon spp. (median seven parasites per 100 leukocytes; range 1-250). Significant relationships were not observed between numbers of leukocytes and infection intensities of Hepatozoon spp. Among the possible explanations for these null results are that Hepatozoon spp. is benign, that Hepatozoon spp. is able to evade detection by the immune system, that Hepatozoon spp. is able to manipulate leukocyte investment, or that other unmeasured or undetected parasites were more important in affecting immune response.
Waggener, W L; Carroll, E J
1998-02-01
Injections of synthetic human gonadotropin releasing hormone (GnRH) into the dorsal pelvic area were used in an attempt to stimulate sperm release in isolated males of eight anuran species including Xenopus laevis, Rana pipiens and Lepidobatrachus laevis. Sperm were obtained within 1-5 h post injection either by mechanical stimulation or by cloacal lavage. Sperm suspensions varied from 8 microL to 7 mL and the cell densities ranged from 4 x 10(5) to 4 x 10(7) sperm/mL. The sperm obtained from seven species using GnRH-induced release were viable based on light microscopic observations of motility. In addition, sperm preparations fertilized eggs in vitro and produced normal tadpoles in the case of L. laevis and L. Ilanensis. This hormonal method of anuran sperm collection will provide a convenient non-injurious way to obtain anuran sperm for basic studies of reproduction and development.
Hossack, B.R.; Corn, P.S.; Pilliod, D.S.
2005-01-01
We surveyed 88 upland wetlands and 12 1-km river sections for amphibians in Theodore Roosevelt National Park, North Dakota, during 2001–2002 to gather baseline data for future monitoring efforts and to evaluate changes in the distribution of species. We compared our results to collections of herpetofauna made during 1920–1922, 1954 and 1978–1979. The boreal chorus frog (Pseudacris maculata) was the most common amphibian in upland wetlands, followed by the tiger salamander (Ambystoma tigrinum), Woodhouse's toad (Bufo woodhousii), northern leopard frog (Rana pipiens), plains spadefoot (Spea bombifrons) and the Great Plains toad (B. cognatus). Bufo woodhousii was the only species that bred in the river. Our records for reptiles are less complete than for amphibians but no losses from the community are evident. The herpetofauna in Theodore Roosevelt National Park seems unchanged during at least the last half-century and likely since 1920–1922.
Knopp, T; Merilä, J
2009-02-01
The postglacial history of the moor frog (Rana arvalis) in Northern Europe was investigated with the aid of eight variable microsatellite loci and a 661 bp sequence of the mitochondrial cytochrome b gene. A division between eastern and western mitochondrial lineages was discovered, supporting two recolonization routes to Fennoscandia since the last glacial maximum. This result was corroborated by the microsatellite data, which revealed a contact zone between the two lineages in Northern Sweden. These findings add to the increasing evidence that an intraspecific genetic biodiversity founded on the existence of eastern and western clades is a common element in Fennoscandian fauna and flora.
Flow cytometry used to assess genetic damage in frogs from farm ponds
Bly, B.L.; Knutson, M.G.; Sandheinrich, M.B.; Gray, B.R.; Jobe, D.A.
2004-01-01
Flow cytometry (FC) is a laboratory method used to detect genetic damage induced by environmental contaminants and other stressors in animals, including amphibians. We tested FC methods on three species of ranid frogs collected from farm ponds and natural wetlands in southeastern Minnesota. We compared FC metrics for Rana clamitans between ponds with direct exposure to agricultural contaminants and reference (unexposed) ponds. Concentrations of atrazine in water from our farm ponds ranged from 0.04 to 0.55 ppb. We found that R. clamitans from exposed ponds had DNA content similar to frogs from unexposed ponds. Pond-averaged C-values (a measure of DNA content) ranged from 6.53 to 7.08 for R. pipiens (n . 13), 6.55 to 6.60 for R. clamitans (n . 40) and 6.74 for R. palustris (n . 5). Among all species, the mean sample CVs ranged from 1.91 (R. palustris) to 6.31 (R. pipiens). Deformities were observed in only 2 of 796 individuals among all species and occurred in both reference and exposed ponds. Although we did not detect evidence of DNA damage associated with agriculture in our study, we demonstrated the potential of FC for screening amphibian populations for genetic damage. Metrics from a variety of amphibian species and locations as well as laboratory studies are needed to further assess the value of FC for monitoring amphibian genetic integrity in contaminated sites.
Richards, Stephanie L; Lord, Cynthia C; Pesko, Kendra N; Tabachnick, Walter J
2010-07-01
Interactions between environmental and biological factors affect the vector competence of Culex pipiens quinquefasciatus for West Nile virus. Three age cohorts from two Cx. p. quinquefasciatus colonies were fed blood containing a low- or high-virus dose, and each group was held at two different extrinsic incubation temperatures (EIT) for 13 days. The colonies differed in the way that they responded to the effects of the environment on vector competence. The effects of mosquito age on aspects of vector competence were dependent on the EIT and dose, and they changed depending on the colony. Complex interactions must be considered in laboratory studies of vector competence, because the extent of the genetic and environmental variation controlling vector competence in nature is largely unknown. Differences in the environmental (EIT and dose) and biological (mosquito age and colony) effects from previous studies of Cx. p. quinquefasciatus vector competence for St. Louis encephalitis virus are discussed.
Ren, Xiaojie; Fu, Shihong; Dai, Peifang; Wang, Huanyu; Li, Yuanyuan; Li, Xiaolong; Lei, Wenwen; Gao, Xiaoyan; He, Ying; Lv, Zhi; Cheng, Jingxia; Wang, Guiqin; Liang, Guodong
2017-06-08
The increasing trend of adult cases of Japanese encephalitis (JE) in China, particularly in northern China, has become an important public health issue. We conducted an epidemiological investigation in the south of Shanxi Province to examine the relationships between mosquitoes, Japanese encephalitis virus (JEV), and adult JE cases. Mosquito specimens were collected from the courtyards of farmers' households and pig farms in Shanxi Province. Mosquitoes were pooled, homogenized, and centrifuged. Reverse transcription-polymerase chain reaction (RT-PCR) was used to detect mosquito-borne arbovirus genes in homogenates. Specimens positive for these genes were inoculated into the baby hamster kidney cell line (BHK-21) to isolate virus. Minimum infection rate was calculated and phylogenetic analyses were performed. A total of 7 943 mosquitoes belonging to six species in four genera were collected; Culex tritaeniorhynchus accounted for 73.08% (5 805/7 943), C. pipiens pallens for 24.75% (1 966/7 943), and the remaining 3% (104/ 7943) consisted of Anopheles sinensis, Aedes vexans, Ae. dorsalis, and Armigeres subalbatus. Sixteen pools were positive for JEV based on RT-PCR using JEV pre-membrane gene nested primers. Phylogenetic analyses showed that all JEVs belonged to genotype I; two pools were positive using Getah Virus (GETV) gene primers. In addition, one JEV strain (SXYC1523) was isolated from C. pipiens pallens specimens. These results indicate that the minimum infection rate of JEV in mosquito specimens collected from the courtyards of farmers' households with pigsties was 7.39/1 000; the rate for pig farms was 2.68/1 000; and the rate for farmers' courtyards without pigsties was zero. The high-prevalence regions of adult JE investigated in this study are still the natural epidemic focus of JEV. Having pigsties near dwellings is a potential risk factor contributing to the prevalence of adult JE. To prevent the occurrence of local adult JE cases, a recommendation was
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...
Anderson, C W
2001-09-01
Using injections of small molecular weight fluorescein dextran amines, combined with activity-dependent uptake of sulforhodamine 101 (SR101), brainstem circuits presumed to be involved in feeding motor output were investigated. As has been shown previously in other studies, projections to the cerebellar nuclei were identified from the cerebellar cortex, the trigeminal motor nucleus, and the vestibular nuclei. Results presented here suggest an additional pathway from the hypoglossal motor nuclei to the cerebellar nucleus as well as an afferent projection from the peripheral hypoglossal nerve to the Purkinje cell layer of the cerebellar cortex. Injections in the cerebellar cortex combined with retrograde labeling of the peripheral hypoglossal nerve demonstrate anatomical convergence at the level of the medial reticular formation. This suggests a possible integrative region for afferent feedback from the hypoglossal nerve and information through the Purkinje cell layer of the cerebellar cortex. The activity-dependent uptake of SR101 additionally suggests a reciprocal, polysynaptic pathway between this same area of the medial reticular formation and the trigeminal motor nuclei. The trigeminal motor neurons innervate the m adductor mandibulae, the primary mouth-closing muscle. The SR101 uptake clearly labeled the ventrolateral hypoglossal nuclei, the medial reticular formation, and the Purkinje cell layer of the cerebellar cortex. Unlike retrograde labeling of the peripheral hypoglossal nerve, stimulating the hypoglossal nerve while SR101 was bath-applied labeled trigeminal motor neurons. This, combined with the dextran labeling, suggests a reciprocal connection between the trigeminal motor nuclei and the cerebellar nuclei, as well as the medulla. Taken together, these data are important for understanding the neurophysiological pathways used to coordinate the proper timing of an extremely rapid, goal-directed movement and may prove useful for elucidating some of the first principles of sensorimotor integration.
Richards, Stephanie L; Lord, Cynthia C; Pesko, Kendra; Tabachnick, Walter J
2009-08-01
Complex interactions between environmental and biological factors influence the susceptibility of Culex pipiens quinquefasciatus to St. Louis encephalitis virus and could affect the epidemiology of virus transmission. Similar interactions could have epidemiologic implications for other vector-virus systems. We conducted an experiment to examine four such factors in combination: mosquito age, extrinsic incubation temperature (EIT), virus dose, and colony. The proportion of mosquitoes with body infections or disseminated infections varied between colonies, and was dependant on age, EIT, and dose. We also show that the probability of a body or leg infection interacted in complex ways between colonies, ages, EITs, and doses. The complex interactive effects of environmental and biological factors must be taken into account for studies of vector competence and epidemiology, especially when laboratory studies are used to generalize to natural transmission dynamics where the extent of variation is largely unknown.
Zink, Steven D; Van Slyke, Greta A; Palumbo, Michael J; Kramer, Laura D; Ciota, Alexander T
2015-10-27
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 inWNV-exposed and/or infected mosquitoes, yet the most consistent and notable was the genus Serratia.
Isolation and characterization of Vasa in the frog Rana rugosa.
Saotome, Kazuhiro; Hayashi, Kota; Adachi, Noritaka; Nakamura, Yoriko; Nakamura, Masahisa
2010-08-01
We cloned a cDNA encoding Vasa, a member of the DEAD (Asp-Glu-Ala-Asp) family of proteins, from the ovary of the frog Rana rugosa. Comparative alignment of amino acid sequences with known Vasa from several species of vertebrate showed that the R. rugosa orthologue shares eight conserved regions with Vasa from other vertebrates. Vasa gene expression was restricted to the testis and ovary among ten different tissues examined. Vasa expression showed no sexual dimorphism during sex determination in R. rugosa, but became higher in the ovary thereafter. By Western blot analysis, a single Vasa band with a molecular weight of 80.9 kDa was detected. The same antibody immunohistochemically detected Vasa in a few cells in the embryonic endoderm at stage 15; the beginning of closure of neural folds, and in the cytoplasm of spermatogonia in the testis, and oocytes in the ovary of tadpoles at stage XX; marked by one or both forelegs protruded. Together, these results suggest that Vasa is a highly specific marker of germ cells and hence useful for studies of germ cell specification and function in amphibians as it already is in other species of both invertebrates and vertebrates such as Drosophila and zebrafish. (c) 2010 Wiley-Liss, Inc.
Pearl, Christopher A.; Adams, Michael J.; Wente, Wendy
2007-01-01
Several western ranid frogs possess a unique strategy of breeding communally over a short temporal window and reusing oviposition sites between years. However, little is published on the characteristics of oviposition sites selected by these explosive breeders. The Columbia spotted frog (Rana luteiventris) is native to northwestern North America and is of conservation concern in the southern portions of its range. As part of a study examining relationships between livestock grazing and R. luteiventris habitat, we assessed characteristics of the species' oviposition sites in 25 fishless ponds in northeastern Oregon. Oviposition sites were generally in shallow water (<25 cm) close to shore and tended to be in the northeastern portion of ponds. Oviposition sites were found more frequently over heavily vegetated substrates and in areas of less substrate slope and shade than random points in littoral zones. We did not quantify temperature differences within ponds, but the patterns we documented are consistent with preferential use of warmer microhabitats for oviposition.
Long-term pathogenic response to Plasmodium relictum infection in Culex pipiens mosquito.
Pigeault, Romain; Villa, Manon
2018-01-01
The transmission of Plasmodium within a vertebrate host population is strongly associated with the life history traits of its vector. Therefore the effect of malaria infection on mosquito fecundity and longevity has traditionally received a lot of attention. Several species of malaria parasites reduce mosquito fecundity, nevertheless almost all of the studies have focused only on the first gonotrophic cycle. Yet, during their lifetime, female mosquitoes go through several gonotrophic cycles, which raises the question of whether they are able to compensate the fecundity costs induced by the parasite. The impact of Plasmodium infection on female longevity is not so clear and has produced conflicting results. Here we measured the impact of Plasmodium relictum on its vector's longevity and fecundity during three consecutive gonotrophic cycles. In accordance with previous studies, we observed a negative impact of Plasmodium infection on mosquito (Culex pipiens) fecundity in the first gonotrophic cycle. Interestingly, despite having taken two subsequent uninfected blood meals, the negative impact of malaria parasite persisted. Nevertheless no impact of infection on mosquito longevity was observed. Our results are not in line with the hypothesis that the reduction of fecundity observed in infected mosquitoes is an adaptive strategy of Plasmodium to increase the longevity of its vector. We discuss the different underlying mechanisms that may explain our results.
Richards, Stephanie L.; Lord, Cynthia C.; Pesko, Kendra; Tabachnick, Walter J.
2009-01-01
Complex interactions between environmental and biological factors influence the susceptibility of Culex pipiens quinquefasciatus to St. Louis encephalitis virus and could affect the epidemiology of virus transmission. Similar interactions could have epidemiologic implications for other vector-virus systems. We conducted an experiment to examine four such factors in combination: mosquito age, extrinsic incubation temperature (EIT), virus dose, and colony. The proportion of mosquitoes with body infections or disseminated infections varied between colonies, and was dependant on age, EIT, and dose. We also show that the probability of a body or leg infection interacted in complex ways between colonies, ages, EITs, and doses. The complex interactive effects of environmental and biological factors must be taken into account for studies of vector competence and epidemiology, especially when laboratory studies are used to generalize to natural transmission dynamics where the extent of variation is largely unknown. PMID:19635881
Trujillo, Xóchitl; Sánchez-Pastor, Enrique; Andrade, Felipa; Huerta, Miguel
2014-01-01
We investigated the effects of cannabinoids on acetylcholine (ACh) or choline contractures in slow skeletal muscle fibers from Rana pipiens. Bundles of cruralis muscle fibers were incubated with the cannabinoid receptor 1 (CB1) agonist, arachidonylcyclopropylamide (ACPA), which diminished the maximum isometric tension by 10 % and the total tension by 5 % of the ACh contracture, and 40 and 22 % of the choline contracture, respectively. Preincubation with the CB1 antagonist, AM281, or with pertussis toxin (PTX) completely blocked the effect of ACPA on the ACh contracture. On the other hand, the decrease in choline contracture by ACPA was only partially blocked by AM281 (~16 % decrease), PTX (20 %), or by dantrolene (~46 %). Our results show that ACPA modulates ACh and choline contractures, and suggest that this effect involves the participation of CB1, the ACh receptor, and -RyR in ACh contractures. For choline contractures, ACPA may also be acting through cannabinoid receptor-independent mechanisms.
Comparative studies on ecotoxicology of synthetic detergents.
Lal, H; Misra, V; Viswanathan, P N; Krishna Murti, C R
1983-12-01
To predict the comparative toxicological response of synthetic detergents on aquatic ecosystems, the effects of various concentrations of neutralized alkyl benzene sulfonate were studied. The median tolerance limit at 48 hr, 95% confidence limit, slope function, presumable harmless concentration, and rate of survival of different species of aquatic fauna such as water fleas (Daphnia magna), mosquito larvae (Culex pipiens), slug worms (Tubifex rivulorum), snails (Lymnaea vulgaris), tadpoles (Rana cyanophlyctis), and fish fingerlings (Cirrhina mrigala) were followed at 0, 24, 48, 72, and 96 hr. Any effect on quality of the water was also tested after the addition of various concentrations of detergents. The results showed that water fleas are more susceptible to detergent toxicity than fish fingerlings, tadpoles, slug worms, snails, and mosquito larvae. Behavioral changes were also observed as an index for detergent toxicity. The relative toxicity of the detergents to various species is discussed in relation to selective ecotoxicological response.
Sweeney, Lauren J; Brodfuehrer, Peter D; Raughley, Beth L
2004-12-01
One important goal of introductory biology laboratory experiences is to engage students directly in all steps in the process of scientific discovery. Even when laboratory experiences are built on principles discussed in the classroom, students often do not adequately apply this background to interpretation of results they obtain in lab. This disconnect has been described at the level of medical education (4), so it should not be surprising that educators have struggled with this same phenomenon at the undergraduate level. We describe a new introductory biology lab that challenges students to make these connections. The lab utilizes enzyme histochemistry and morphological observations to draw conclusions about the composition of functionally different types of muscle fibers present in skeletal muscle. We report that students were not only successful at making these observations on a specific skeletal muscle, the gastrocnemius of the frog Rana pipiens, but that they were able to connect their results to the principles of fiber type differences that exist in skeletal muscles in all vertebrates.
Fatigue in isometric contraction in a single muscle fibre: a compartmental calcium ion flow model.
Kothiyal, K P; Ibramsha, M
1986-01-01
Fatigue in muscle is a complex biological phenomenon which has so far eluded a definite explanation. Many biochemical and physiological models have been suggested in the literature to account for the decrement in the ability of muscle to sustain a given level of force for a long time. Some of these models have been critically analysed in this paper and are shown to be not able to explain all the experimental observations. A new compartmental model based on the intracellular calcium ion movement in muscle is proposed to study the mechanical responses of a muscle fibre. Computer simulation is performed to obtain model responses in isometric contraction to an impulse and a train of stimuli of long duration. The simulated curves have been compared with experimentally observed mechanical responses of the semitendinosus muscle fibre of Rana pipiens. The comparison of computed and observed responses indicates that the proposed calcium ion model indeed accounts very well for the muscle fatigue.
Zheng, W; Hall, J C
2000-01-01
The role of gamma-aminobutyric acid (GABA)ergic inhibition in shaping the excitatory frequency tuning of 74 neurons in the superior olivary nucleus of the leopard frog, Rana pipiens, was studied using iontophoretic application of the GABA(A) receptor antagonist, bicuculline methiodide. For 37 neurons, bicuculline application broadened and/or changed the configuration of the excitatory frequency-tuning curve. Results indicate that GABA-mediated inhibition not only sharpens the tuning curves of neurons but also plays a critical role in creating new frequency tuning properties in the superior olivary nucleus. Bicuculline application affected other neuronal response properties as well. Spontaneous firing rate increased 11-338% for 18 of 59 neurons. For 32 of 58 neurons there was an increase in stimulus-evoked discharge rate and a change in rate-level function. There was no qualitative effect on the discharge pattern of 60 neurons, though 2 tonically responding neurons did show an increase (> 30%) in response duration. Additional roles for GABAergic inhibition in monaural signal analysis are discussed.
Species accounts for the Alamosa/Monte Vista/Baca National Wildlife Refuge Complex
Ellison, Laura E.
2011-01-01
As part of an interagency agreement between the U.S. Geological Survey (USGS) and the U.S. Fish and Wildlife Service (USFWS), the Alamosa/Monte Vista/Baca National Wildlife Refuge Complex requested help with the synthesis of scientific information for 10 focal species and their habitat requirements in response to common Refuge management activities in the San Luis Valley, Colorado. This information will be instrumental in developing the Service's Comprehensive Conservation Plan (CCP), which is required by law for each unit of the National Wildlife Refuge System. After consultation with Refuge managers and USGS staff, the 10 species chosen for detailed literature reviews and synthesis of information were the following: (1) American Avocet (Recurvirostra americana); (2) Wilson's Phalarope (Phalaropus tricolorPorzana carolina); (4) White-faced Ibis (Plegadis chihi); (5) Black Tern (Chlidonias niger); (6) Short-eared Owl (Asio flammeus); (7) Brewer's Sparrow (Spizella breweri); (8) Savannah Sparrow (Passerculus sandwichensis); (9) Northern Leopard Frog [Lithobates (=Rana) pipiens]; and, (10) Tadpole Shrimp (Triops longicaudatus).
Baldacchino, F; Montarsi, F; Arnoldi, D; Barategui, C; Ferro Milone, N; Da Rold, G; Capelli, G; Rizzoli, A
2017-05-01
Aedes koreicus (Edwards) is an invasive mosquito species, like Aedes albopictus (Skuse) and Aedes japonicus japonicus (Theobald), that has already colonized a large part of northeastern Italy and other European countries. Despite its rapid expansion, information about adult distribution and trapping is lacking. Here, we conducted a 2-yr longitudinal survey using adult traps to investigate the spatiotemporal distribution of Ae. koreicus and evaluated the effectiveness of three trapping devices in Latin square experiments conducted in an urban site and a forested site. The following three different traps were compared: a CO2-baited Biogents (BG) Sentinel trap, a CO2-baited Centers for Disease Control and Prevention light trap (CDC trap), and a grass infusion-baited gravid trap.In northern Italy, Ae. koreicus was collected from late April to early November, with peak of abundance observed in August. Aedes koreicus was more abundant in 2015 than in 2014 because of higher temperatures during summer. Unlike Ae. albopictus, the abundance of Ae. koreicus was not related to the altitude of the sampling locations in the range 241-660 m above sea level. The BG Sentinel and gravid traps collected significantly more Ae. koreicus than the CDC trap in the urban site, whereas there was no significant difference between the three traps in the forested site. In the urban site, the BG Sentinel trap and the gravid trap were the most effective for collecting Ae. albopictus and Culex pipiens L., respectively. In the forested site, Cx. pipiens was primarily collected by the CDC trap. © The Authors 2017. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.
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.
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.
Pool desiccation and developmental thresholds in the common frog, Rana temporaria.
Lind, Martin I; Persbo, Frida; Johansson, Frank
2008-05-07
The developmental threshold is the minimum size or condition that a developing organism must have reached in order for a life-history transition to occur. Although developmental thresholds have been observed for many organisms, inter-population variation among natural populations has not been examined. Since isolated populations can be subjected to strong divergent selection, population divergence in developmental thresholds can be predicted if environmental conditions favour fast or slow developmental time in different populations. Amphibian metamorphosis is a well-studied life-history transition, and using a common garden approach we compared the development time and the developmental threshold of metamorphosis in four island populations of the common frog Rana temporaria: two populations originating from islands with only temporary breeding pools and two from islands with permanent pools. As predicted, tadpoles from time-constrained temporary pools had a genetically shorter development time than those from permanent pools. Furthermore, the variation in development time among females from temporary pools was low, consistent with the action of selection on rapid development in this environment. However, there were no clear differences in the developmental thresholds between the populations, indicating that the main response to life in a temporary pool is to shorten the development time.
Climatic influences on the breeding biology of the agile frog ( Rana dalmatina)
NASA Astrophysics Data System (ADS)
Combes, Magali; Pinaud, David; Barbraud, Christophe; Trotignon, Jacques; Brischoux, François
2018-02-01
Severe population declines of amphibians have been shown to be attributed to climate change. Nevertheless, the various mechanisms through which climate can influence population dynamics of amphibians remain to be assessed, notably to disentangle the relative synergetic or antagonistic influences of temperature and precipitations on specific life history stages. We investigated the impact of rainfall and temperature on the egg-clutch abundance in a population of agile frog ( Rana dalmatina) during 29 years (1987-2016) on 14 breeding sites located in Brenne Natural Park, France. Specifically, we examined the influence of environmental conditions occurring during five temporal windows of the year cycle corresponding to specific life history stages. Overall, our results suggest that the year-to-year fluctuations of egg-clutch abundances in Brenne Natural Park were partly dependent on local climatic conditions (rainfall and temperature). Climate seemed to influence breeding frogs during the autumn-winter period preceding reproduction. Spring and summer conditions did not influence reproduction. Additionally, we failed to detect effects of climatic conditions on newly metamorphosed individuals. Other factors such as density dependence and inter-specific interactions with introduced predators are likely to play a significant role in reproduction dynamics of the studied frog populations.
Rana computatrix to human language: towards a computational neuroethology of language evolution.
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.
Climatic influences on the breeding biology of the agile frog (Rana dalmatina).
Combes, Magali; Pinaud, David; Barbraud, Christophe; Trotignon, Jacques; Brischoux, François
2017-12-19
Severe population declines of amphibians have been shown to be attributed to climate change. Nevertheless, the various mechanisms through which climate can influence population dynamics of amphibians remain to be assessed, notably to disentangle the relative synergetic or antagonistic influences of temperature and precipitations on specific life history stages. We investigated the impact of rainfall and temperature on the egg-clutch abundance in a population of agile frog (Rana dalmatina) during 29 years (1987-2016) on 14 breeding sites located in Brenne Natural Park, France. Specifically, we examined the influence of environmental conditions occurring during five temporal windows of the year cycle corresponding to specific life history stages. Overall, our results suggest that the year-to-year fluctuations of egg-clutch abundances in Brenne Natural Park were partly dependent on local climatic conditions (rainfall and temperature). Climate seemed to influence breeding frogs during the autumn-winter period preceding reproduction. Spring and summer conditions did not influence reproduction. Additionally, we failed to detect effects of climatic conditions on newly metamorphosed individuals. Other factors such as density dependence and inter-specific interactions with introduced predators are likely to play a significant role in reproduction dynamics of the studied frog populations.
Workplace safety in Bangladesh ready-made garment sector: 3 years after the Rana Plaza collapse.
Barua, Uttama; Ansary, Mehedi Ahmed
2017-12-01
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.
Effects of lead-contaminated sediment on Rana sphenocephala tadpoles.
Sparling, Donald W; Krest, Sherry; Ortiz-Santaliestra, Manuel
2006-10-01
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 microg/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 microg/L lead in pore water (95% CI = 4000 to 35,200 microg/L). Early growth and development were depressed at 2,360 mg/kg lead in sediment (8100 microg/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.
Effects of lead-contaminated sediment on Rana sphenocephala tadpoles
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.
Bee, M A
2001-09-01
The aggressive response of male bullfrogs (Rana catesbeiana) habituates with repeated broadcasts of acoustic stimuli simulating a new territorial neighbor. The effects of stimulus repetition rate and stimulus intensity on bullfrog aggressive responses were tested in a field experiment designed to test the assumptions of a dual-process theory of habituation. Synthetic advertisement calls were broadcast at 2 repetition rates and 2 intensities in a factorial design. Bullfrogs were more aggressive at the higher stimulus intensity at both repetition rates. Aggressive responses habituated more slowly at the higher stimulus intensity and slower repetition rate compared with other treatments. Several biotic and abiotic factors had small or negligible effects on aggressive responses. Although consistent with the operation of 2 opposing processes, habituation and sensitization, the data provide only partial support for the assumptions of dual-process theory.
Relyea, Rick A
2012-03-01
The use of pesticides is important for growing crops and protecting human health by reducing the prevalence of targeted pest species. However, less attention is given to the potential unintended effects on nontarget species, including taxonomic groups that are of current conservation concern. One issue raised in recent years is the potential for pesticides to become more lethal in the presence of predatory cues, a phenomenon observed thus far only in the laboratory. A second issue is whether pesticides can induce unintended trait changes in nontarget species, particularly trait changes that might mimic adaptive responses to natural environmental stressors. Using outdoor mesocosms, I created simple wetland communities containing leaf litter, algae, zooplankton, and three species of tadpoles (wood frogs [Rana sylvatica or Lithobates sylvaticus], leopard frogs [R. pipiens or L. pipiens], and American toads [Bufo americanus or Anaxyrus americanus]). I exposed the communities to a factorial combination of environmentally relevant herbicide concentrations (0, 1, 2, or 3 mg acid equivalents [a.e.]/L of Roundup Original MAX) crossed with three predator-cue treatments (no predators, adult newts [Notophthalmus viridescens], or larval dragonflies [Anax junius]). Without predator cues, mortality rates from Roundup were consistent with past studies. Combined with cues from the most risky predator (i.e., dragonflies), Roundup became less lethal (in direct contrast to past laboratory studies). This reduction in mortality was likely caused by the herbicide stratifying in the water column and predator cues scaring the tadpoles down to the benthos where herbicide concentrations were lower. Even more striking was the discovery that Roundup induced morphological changes in the tadpoles. In wood frog and leopard frog tadpoles, Roundup induced relatively deeper tails in the same direction and of the same magnitude as the adaptive changes induced by dragonfly cues. To my knowledge, this
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.
Chen, Zhongyuan; Kearney, Christopher M
2015-06-01
We chose five easily propagated garden plants previously shown to be attractive to mosquitoes, ants or other insects and tested them for attractiveness to Culex pipiens and Aedes aegypti. Long term imbibition was tested by survival on each plant species. Both mosquito species survived best on Impatiens walleriana, the common garden impatiens, followed by Asclepias curassavica, Campsis radicans and Passiflora edulis, which sponsored survival as well as the 10% sucrose control. Immediate preference for imbibition was tested with nectar dyed in situ on each plant. In addition, competition studies were performed with one dyed plant species in the presence of five undyed plant species to simulate a garden setting. In both preference studies I. walleriana proved superior. Nectar from all plants was then screened for nectar protein content by SDS-PAGE, with great variability being found between species, but with I. walleriana producing the highest levels. The data suggest that I. walleriana may have value as a model plant for subsequent studies exploring nectar delivery of transgenic mosquitocidal proteins. Copyright © 2015 Elsevier B.V. All rights reserved.
Atrazine increases the sodium absorption in frog (Rana esculenta) skin.
Cassano, Giuseppe; Bellantuono, Vito; Ardizzone, Concetta; Lippe, Claudio
2006-02-01
The presence of atrazine in agricultural sites has been linked to the decline in amphibian populations. The efforts of the scientific community generally are directed toward investigating the long-term effect of atrazine on complex functions (reproduction or respiration), but in the present study, we investigated the short-term effect on the short-circuit current (I(sc)), a quantitative measure of the ion transport operated by frog (Rana esculenta) skin. Treatment with 5 microM atrazine (1.08 mg/L) does not affect the transepithelial outfluxes of [14C]mannitol or [14C]urea; therefore, atrazine does not damage the barrier properties of frog skin. Atrazine causes a dose-dependent increase in the short-circuit current, with a minimum of 4.64 +/- 0.76 microA/cm2 (11.05% +/- 1.22%) and a maximum of 12.7 +/- 0.7 microA/cm2 (35% +/- 2.4%) measured at 10 nM and 5 microM, respectively. An increase in Isc also is caused by 5 microM ametryne, prometryn, simazine, terbuthylazine, or terbutryn (other atrazine derivatives). In particular, atrazine increases the transepithelial 22Na+ influx without affecting the outflux. Finally, stimulation of Isc by atrazine is suppressed by SQ 22536, H89, U73122, 2-aminoethoxydiphenyl borate, and W7 (blockers of adenylate cyclase, protein kinase A, phospholipase C, intracellular Ca2+ increase, and calmodulin, respectively), whereas indomethacin and calphostin C (inhibitors of cyclooxygenase and protein kinase C, respectively) have no effect.
High chlorpyrifos resistance in Culex pipiens mosquitoes: strong synergy between resistance genes
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
Yaw, Taylor; Swanson, Jennifer E; Pierce, Clay; Muths, Erin L.; Smalling, Kelly; Vandever, Mark; Zaffarano, Bianca Anne
2017-01-01
Historically, wetland toxin exposure studies have relied on single time point samples from stationary sampling devices. Development of passive sampling devices (PSDs) that can be attached to individual animals within wetland habitats has greatly improved in recent years, presenting an innovative sampling technology that can potentially yield individual-specific, quantifiable data about chemical exposure. In this study, silicone based PSDs were attached to the ventral skin of 20 northern leopard frogs (Lithobates pipiens) with polypropylene sutures after radio transmitters had been surgically implanted into the coleomic cavity. After a recovery period frogs were released back into the wetland habitat where they were acquired. The animals were located daily using radio telemetry to assess how long PSDs would remain attached in the frogs' natural habitat. After one week, PSDs remained on 18 of the original 20 frogs. At 2 weeks 17 frogs were recovered and no PSDs remained attached. Although valuable data can be obtained over a short time period, more research will be necessary to demonstrate effectiveness of externally attaching silicone PSDs to northern leopard frogs for time periods longer than 1-2 weeks.
Knutson, Melinda G.; Herner-Thogmartin, Jennifer H.; Thogmartin, Wayne E.; Kapfer, Joshua M.; Nelson, John
2018-01-01
Telemetry data for 59 Northern Leopard Frogs (Lithobates pipiens) breeding in ponds in Houston and Winona Counties, MN; 2001-2002. Agricultural intensification is causing declines in many wildlife species, including Northern Leopard Frogs (Lithobates pipiens). Specific information about frog movements, habitat selection, and sources of mortality can be used to inform conservation-focused land management and acquisition. We studied Northern Leopard Frogs in southeastern Minnesota, part of the Driftless Area ecoregion, characterized by hills and valleys and a mix of agriculture, forests, small towns and farmsteads. In this area, small farm ponds, originally built to control soil erosion are used by the species for breeding and wintering in addition to riparian wetlands. But, this agricultural landscape may be hazardous for frogs moving between breeding, feeding, and wintering habitats. We surgically implanted transmitters into the peritoneal cavity of 59 Northern Leopard Frogs and tracked them from May to October 2001-2002. The total distance traveled by radio-tagged frogs ranged from 12 to 3316 m, the 95% home range averaged 5.3 ± 1.2 (SE) ha, and the 50% core area averaged 1.05 ± 0.3 (SE) ha. As expected, Northern Leopard Frogs selected wetlands over all other land cover classes and row crops were generally avoided at all levels of selection. Only a few tracked frogs were successful at dispersing (n = 6). Most frogs attempting to disperse (n =31) ended up missing (n = 14), died due to mowing (n = 8), or were recorded as transmitter failure (n = 2) or unknown mortalities (n = 1). For the conservation of Northern Leopard Frogs in this agricultural setting, we must consider both the aquatic and the terrestrial needs of this species. Conservation agencies that restore, manage, and acquire wetlands should consider the hazards posed by land uses adjacent to frog breeding and wintering sites and plan for movement corridors between these locations. For example
Multiple sublethal chemicals negatively affect tadpoles of the green frog, Rana clamitans
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.
Lilje, Jonathan; Mosler, Hans-Joachim
2018-04-01
Worldwide, an estimated 700 million people rely on unimproved drinking water sources; even more consume water that is not safe to drink. Inadequate drinking water quality constitutes a major risk factor for cholera and other diarrheal diseases around the globe, especially for young children in developing countries. Household water treatment and safe storage systems represent an intermediate solution for settings that lack infrastructure supplying safe drinking water. However, the correct and consistent usage of such treatment technologies rely almost exclusively on the consumer's behavior. This study targeted at evaluating effects of a behavior change campaign promoting the uptake of household drinking water chlorination in communities along the Chari and Logone rivers in Chad. The campaign was based on formative research using health psychological theory and targeted several behavioral factors identified as relevant. A total of 220 primary caregivers were interviewed concerning their household water treatment practices and mindset related to water treatment six months after the campaign. The Risks, Attitudes, Norms, Abilities, and Self-regulation (RANAS) model was used to structure the interviews as the RANAS approach had been used for designing the campaign. Results show significantly higher self-reported drinking water chlorination among participants of the intervention. Significant differences from a control group were identified regarding several behavioral factors. Mediation analysis revealed that the intervention positively affected participants' individual risk estimation for diarrheal disease, health knowledge, perceived efforts and benefits of water treatment, social support strategies, knowledge of how to perform chlorination, and perceived ability to do so. The campaign's effect on water treatment was mainly mediated through differences in health knowledge, changes in norms, and self-efficacy convictions. The findings imply that water treatment behavior
Zhigalev, O N
2010-01-01
The genetic structure of populations of four helminth species from moor frog Rana arvalis, in comparison with the population-genetic structure of the host, has been studied with the gel-electrophoresis method. As compared with the host, parasites are characterized by more distinct deviation from the balance of genotypic frequencies and higher level of interpopulation genetic differences. The genetic variability indices in the three of four frog helminthes examined are lower than those in the host. Moreover, these indices are lower than the average indices typical of free-living invertebrates; this fact contradicts the opinion on polyhostality of these helminthes and their wide distribution.
MiR-285 targets P450 (CYP6N23) to regulate pyrethroid resistance in Culex pipiens pallens.
Tian, Mengmeng; Liu, Bingqian; Hu, Hongxia; Li, Xixi; Guo, Qin; Zou, Feifei; Liu, Xianmiao; Hu, Mengxue; Guo, Juxin; Ma, Lei; Zhou, Dan; Sun, Yan; Shen, Bo; Zhu, Changliang
2016-12-01
MicroRNAs play critical roles in post-transcriptional regulation of gene expression, which participate in the modulation of almost all of the cellular processes. Although emerging evidence indicates that microRNAs are related with antineoplastic drugs resistance, whether microRNAs are responsible for insecticide resistance in mosquitos is poorly understood. In this paper, we found that miR-285 was significantly upregulated in the deltamethrin-resistant strain of Culex pipiens pallens, and overexpression miR-285 through microinjection increased mosquito survival rate against deltamethrin treatement. Using bioinformatic software, quantitative reverse transcription PCR, luciferase reporter assay and microinjection approaches, we conformed that CYP6N23 was the target of miR-285. Lower expression of CYP6N23 was observed in the deltamethrin-resistant strain. While, mosquito mortality rate was decreased after downregulating expression of CYP6N23 by dsRNA against CYP6N23 or miR-285 mimic microinjection. These findings revealed that miR-285 could target CYP6N23 to regulate pyrethroid resistance, providing new insights into mosquito insecticide resistance surveillance and control.
Mangiamele, Lisa A; Fuxjager, Matthew J; Schuppe, Eric R; Taylor, Rebecca S; Hödl, Walter; Preininger, Doris
2016-05-17
Physical gestures are prominent features of many species' multimodal displays, yet how evolution incorporates body and leg movements into animal signaling repertoires is unclear. Androgenic hormones modulate the production of reproductive signals and sexual motor skills in many vertebrates; therefore, one possibility is that selection for physical signals drives the evolution of androgenic sensitivity in select neuromotor pathways. We examined this issue in the Bornean rock frog (Staurois parvus, family: Ranidae). Males court females and compete with rivals by performing both vocalizations and hind limb gestural signals, called "foot flags." Foot flagging is a derived display that emerged in the ranids after vocal signaling. Here, we show that administration of testosterone (T) increases foot flagging behavior under seminatural conditions. Moreover, using quantitative PCR, we also find that adult male S. parvus maintain a unique androgenic phenotype, in which androgen receptor (AR) in the hind limb musculature is expressed at levels ∼10× greater than in two other anuran species, which do not produce foot flags (Rana pipiens and Xenopus laevis). Finally, because males of all three of these species solicit mates with calls, we accordingly detect no differences in AR expression in the vocal apparatus (larynx) among taxa. The results show that foot flagging is an androgen-dependent gestural signal, and its emergence is associated with increased androgenic sensitivity within the hind limb musculature. Selection for this novel gestural signal may therefore drive the evolution of increased AR expression in key muscles that control signal production to support adaptive motor performance.
Mangiamele, Lisa A.; Fuxjager, Matthew J.; Schuppe, Eric R.; Taylor, Rebecca S.; Hödl, Walter; Preininger, Doris
2016-01-01
Physical gestures are prominent features of many species’ multimodal displays, yet how evolution incorporates body and leg movements into animal signaling repertoires is unclear. Androgenic hormones modulate the production of reproductive signals and sexual motor skills in many vertebrates; therefore, one possibility is that selection for physical signals drives the evolution of androgenic sensitivity in select neuromotor pathways. We examined this issue in the Bornean rock frog (Staurois parvus, family: Ranidae). Males court females and compete with rivals by performing both vocalizations and hind limb gestural signals, called “foot flags.” Foot flagging is a derived display that emerged in the ranids after vocal signaling. Here, we show that administration of testosterone (T) increases foot flagging behavior under seminatural conditions. Moreover, using quantitative PCR, we also find that adult male S. parvus maintain a unique androgenic phenotype, in which androgen receptor (AR) in the hind limb musculature is expressed at levels ∼10× greater than in two other anuran species, which do not produce foot flags (Rana pipiens and Xenopus laevis). Finally, because males of all three of these species solicit mates with calls, we accordingly detect no differences in AR expression in the vocal apparatus (larynx) among taxa. The results show that foot flagging is an androgen-dependent gestural signal, and its emergence is associated with increased androgenic sensitivity within the hind limb musculature. Selection for this novel gestural signal may therefore drive the evolution of increased AR expression in key muscles that control signal production to support adaptive motor performance. PMID:27143723
Feinberg, Jeremy A.; Newman, Catherine E.; Watkins-Colwell, Gregory J.; Schlesinger, Matthew D.; Zarate, Brian; Curry, Brian R.; Shaffer, H. Bradley; Burger, Joanna
2014-01-01
We describe a new cryptic species of leopard frog from the New York City metropolitan area and surrounding coastal regions. This species is morphologically similar to two largely parapatric eastern congeners, Rana sphenocephala and R. pipiens. We primarily use bioacoustic and molecular data to characterize the new species, but also examine other lines of evidence. This discovery is unexpected in one of the largest and most densely populated urban parts of the world. It also demonstrates that new vertebrate species can still be found periodically even in well-studied locales rarely associated with undocumented biodiversity. The new species typically occurs in expansive open-canopied wetlands interspersed with upland patches, but centuries of loss and impact to these habitats give some cause for conservation concern. Other concerns include regional extirpations, fragmented extant populations, and a restricted overall geographic distribution. We assign a type locality within New York City and report a narrow and largely coastal lowland distribution from central Connecticut to northern New Jersey (based on genetic data) and south to North Carolina (based on call data). PMID:25354068
el-Matbouli, M; Hoffmann, R W; Schoel, H; McDowell, T S; Hedrick, R P
1999-01-07
Scanning electron microscopic studies were conducted on rainbow trout Oncorhynchus mykiss in the first 60 min after their exposure to the triactinomyxon spores of Myxobolus cerebralis. The results demonstrated that as early as 1 min post exposure the whole process, from the attachment of the triactinomyxon spores to the complete penetration of their sporoplasm germs, had occurred. The triactinomyxon spores sought out the secretory openings of mucous cells of the epidermis, the respiratory epithelium and the buccal cavity of trout and used them as portals of entry. Exposure experiments of the triactinomyxon spores of M. cerebralis to non-salmonid fish, such as goldfish Carassius auratus, carp Cyprinus carpio, nose Chondrostoma nasus, medaka Oryzias latipes, guppy Poecilia reticulata and also the amphibian tadpole Rana pipiens as well as to rainbow trout fry indicated a specificity for salmonids. Attempts to activate the triactinomyxon spores by exposure to mucus prepared from cyprinid and salmonid fish showed no significant differences from those conducted in tap water. The results suggest that the simultaneous presence of both mechano- and chemotactic stimuli was required for finding the salmonid fish host.
Sensitivity of diagnostic techniques in determining the prevalence of anuran trypanosomes.
Woo, P T
1983-01-01
Two hundred thirty-three leopard frogs (Rana pipiens) from Oshkosh, Wisconsin, USA, were divided into two groups and their blood examined for trypanosomes. In Group I (n = 157), where the blood was examined by the hematocrit centrifuge technique (HCT), 36 (23%) were infected with trypanosomes. Eighteen were infected with Trypanosoma pipientis, 13 with Trypanosoma ranarum, three with Trypanosoma rotatorium and two with mixed infections of T. pipientis and T. ranarum. In Group II (n = 76) the blood was cultured and also examined by HCT and wet mounts. Trypanosomes (T. pipientis and T. ranarum) were detected in 24 frogs (33%) using all three techniques. Eleven T. pipientis were detected by HCT, however none by culture and two by wet mounts. Twelve T. ranarum were detected by culture while only 10 were found by HCT and five by wet mounts. One T. ranarum infection detected by HCT was missed by culture because of bacterial contamination. The HCT was consistently better than wet mount examinations. It is suggested that the HCT be used whenever possible in future trypanosome surveys.
Apomorphine effects on frog locomotor behavior.
Chu, Joanne; Wilczynski, Walter
2007-05-16
The neuroanatomical pathways of the DA systems have been shown to be largely conserved across many vertebrate taxa. It is less certain whether the structural similarities seen between mammals and amphibians reflect a similar functional homology. DA is well known for its role in facilitating motor behaviors in mammals. We examined whether a similar role for DA exists in amphibians using the Northern Leopard Frog (Rana pipiens). We investigated the effects of the nonspecific DA agonist, apomorphine (APO) on a complex motor task that included two distinct components known to be differentially modulated by DA in mammals: swimming and climbing. We demonstrated that a high single dose of APO (20 mg/kg, body weight) strongly increased the amount of time spent completing the motor task. Furthermore, we showed that although APO did not significantly alter several aspects of swimming behavior, two aspects of climbing behavior were disrupted. Both climbing speed and climbing ability were impaired by APO treatment. These results increase our understanding of DA function in amphibians and add to our understanding of structure-function homologies of dopamine function across vertebrate taxa.
Vershinin, V L
2008-01-01
Under investigation is a complex of inherited physiological properties of the morpha striata (a monogenous dominant mutation) in two species of the genus Rana. Insufficient effectiveness of the potassium-sodium pump responsible for the skin transport in amphibians had lead to formation of a number of compensative physiological mechanisms in this morpha. The yearlings of the morpha striata are characterized by highly dynamic hemopoetic system playing important role in individual adaptations to unstable environments. Such a high level of metabolism in the morpha striata promotes rising of adaptive potential of the nervous system due to decrease of the excitability threshold, but causes shortening the life span. Therefore, physiological differences correlated with polymorph structure of the close species can be of crucial importance in their adaptations under existence in the natural and artificial geochemical anomalies and in anthropogenically disturbed ecosystems.
Origgi, F C; Schmidt, B R; Lohmann, P; Otten, P; Akdesir, E; Gaschen, V; Aguilar-Bultet, L; Wahli, T; Sattler, U; Stoffel, M H
2017-07-01
Amphibian pathogens are of current interest as contributors to the global decline of amphibians. However, compared with chytrid fungi and ranaviruses, herpesviruses have received relatively little attention. Two ranid herpesviruses have been described: namely, Ranid herpesvirus 1 (RHV1) and Ranid herpesvirus 2 (RHV2). This article describes the discovery and partial characterization of a novel virus tentatively named Ranid herpesvirus 3 (RHV3), a candidate member of the genus Batrachovirus in the family Alloherpesviridae. RHV3 infection in wild common frogs (Rana temporaria) was associated with severe multifocal epidermal hyperplasia, dermal edema, a minor inflammatory response, and variable mucous gland degeneration. Intranuclear inclusions were numerous in the affected epidermis together with unique extracellular aggregates of herpesvirus-like particles. The RHV3-associated skin disease has features similar to those of a condition recognized in European frogs for the last 20 years and whose cause has remained elusive. The genome of RHV3 shares most of the features of the Alloherpesviruses. The characterization of this presumptive pathogen may be of value for amphibian conservation and for a better understanding of the biology of Alloherpesviruses.
Effects of fluoride on development and growth of Rana chensinensis embryos and larvae.
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. Copyright © 2015 Elsevier Inc. All rights reserved.
Oral chytridiomycosis in the mountain yellow-legged frog (Rana muscosa)
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.
Oral chytridiomycosis in the mountain yellow-legged frog (Rana muscosa)
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.
The Sound Broadcasting System of the Bullfrog
NASA Astrophysics Data System (ADS)
Purgue, Alejandro P.
1995-01-01
This work presents a comparison across selected species of several aspects of the mechanism of sound broadcasting in anuran amphibians. These studies indicate that all anuran species studied to date broadcast their calls through structures that resonate at the dominant frequency in their calls. Measurements of the magnitude of the transfer function of the radiating structures show that the structures responsible for radiating the bulk of the energy present in the call vary depending on the species considered. Bullfrogs (Rana catesbeiana) radiate most of the energy (89% sound level) present in their calls through their eardrums. In this species the transfer function of the eardrum displays several peaks coincident in frequency and amplitude with the energy distribution observed in the mating and release call of the species. The vocal sac and gular area contribute energy only in the lower band (150 to 400 Hz) of the call. The ears are responsible for radiating additional frequency bands to the ones being radiated through the gular area and vocal sacs. This condition appears to be derived. In Rana pipiens the ears also broadcast a significant portion of the energy present in the call (63% sound level) but the frequencies of the aural emissions are a subset of those frequencies radiated through the vocal sac and gular area. Character optimization suggests that this is the primitive condition for ranid frogs. Finally, the barking treefrog (Hyla gratiosa) appears to use two different structures to radiate different portions of the call. The low frequency band appears to be preferentially radiated through the lungs while the high frequency components of the call are radiated through the vocal sac.
Chytridiomycosis widespread in Anurans of Northeastern United States
Longcore, J.R.; Longcore, J.E.; Pessier, Allan P.; Halteman, W.A.
2007-01-01
An emerging disease of amphibians caused by the chytrid fungus Batrachochytrium dendrobatidis has been associated with morbidity, mortality, and extinction of species. Typically, researchers have detected B. dendrobatidis only when examining amphibians for causes of mortalities; few data exist on infection rates where mortalities are lacking. During May?September 2000?2002 we obtained amphibian specimens killed by vehicles and others collected at remote off-road sites throughout Maine, USA, and from federal lands in 5 states in the Northeast. We detected infected specimens, mostly green frogs (Rana clamitans), at 5 of 7 national wildlife refuges, a federal waterfowl production area, and Acadia National Park. Seven of 9 species, including all Ranidae species, were infected throughout Maine; rates ranged from 14.6% in American toads (Bufo americanus) to 25.7% in northern leopard frogs (Rana pipiens). We did not detect any infections in 50 eastern gray tree frogs (Hyla versicolor) or 21 spring peepers (Pseudacris crucifer). Species that hibernate in terrestrial habitats seem to have lower rates of infection than species that hibernate in aquatic habitats. Infections peaked in spring and autumn and were associated with air temperatures optimal for B. dendrobatidis growth. The relatively high infection rates among species without documented die-offs suggest that either losses have occurred undetected, that the fungus is endemic and species have attained a level of resistance to infections becoming lethal, or that climatic conditions of the Northeast have a role in preventing infections from being lethal. Data on prevalence and distribution of this chytrid fungus in the Northeast may be useful in modeling its origins and predicting long-term ecosystem effects involving anurans.
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.
Kim, Hyunwoo; Cha, Go-Woon; Jeong, Young Eui; Lee, Wook-Gyo; Chang, Kyu Sik; Roh, Jong Yul; Yang, Sung Chan; Park, Mi Yeoun; Park, Chan; Shin, E-Hyun
2015-01-01
Japanese encephalitis virus (JEV) causes significant viral encephalitis and is distributed throughout the Asian countries. The virus is known to be transmitted by Culex tritaeniorhynchus, which mainly breeds in rice paddies in Korea. In this study, we investigated the presence of other mosquito species that can transmit JEV as a second or regional vector. We selected five cities where patients have experienced JE in the last 5 years as mosquito-collecting locations and subdivided them into four collection sites according to the mosquito habitats (cowshed, downtown area, forest, and swamp). Mosquitoes were caught using the BG-Sentinel trap, CDC black-light trap, Fay-Prince trap, and Gravid trap. A total of 993 pools from 22,774 mosquitoes were prepared according to their species, collection date, and site. We performed a SYBR Green 1-based real-time RT-PCR assay to detect JEV from the mosquito pools. A total of six JEV-positive pools were detected from Culex orientalis and Culex pipiens caught in the Gangwon-do and Gyeonngi-do provinces. All the detected JEVs were revealed as genotype V by phylogenetic analysis of the envelope gene. Our findings confirm that a new genotype of JEV was introduced in Korea and suggest that two mosquito species may play a role in JEV transmission.
Curtis, Amanda N; Bidart, M Gabriela
2017-11-01
Invasive plants impact amphibians by altering habitat, altering species interactions, and releasing potentially toxic secondary chemicals. Despite being costly and having the potential to affect nontarget wildlife, chemical management is commonly used to control invasive plants. Prior research has indicated that individual effects of invasive plants or herbicides can be harmful to aquatic organisms; however, information is lacking on the combined effect of these factors on amphibians. A laboratory experiment was performed to assess the impact of leachates of the invasive plants Eurasian watermilfoil (Myriophyllum spicatum) and European buckthorn (Rhamnus cathartica), the herbicide Renovate ® 3 (triclopyr [3, 5, 6-trichloro-2-pyridinyloxyacetic acid]), and the combined effects of each plant leachate and the herbicide on the growth, morphology, and survival of northern leopard frog (Lithobates pipiens) tadpoles. No effects of treatment on survival were observed. Tadpole exposure to M. spicatum reduced body mass by 17%, exposure to R. cathartica increased body mass by 36%, and exposure to R. cathartica + low herbicide increased body mass by 38% (although only early in the experiment). Exposure to Renovate 3 induced a 16% and 29% decrease in tadpole size in lower (0.22 mg triclopyr active ingredient [a.i.]/L) and higher (0.92 mg triclopyr a.i./L) concentration treatments, respectively. Results from the present study highlight the importance of considering both individual and combined effects of invasive plants and herbicides because they may have different outcomes for tadpole growth and development. Environ Toxicol Chem 2017;36:2958-2964. © 2017 SETAC. © 2017 SETAC.
GENE-dosage effects on fitness in recent adaptive duplications: ace-1 in the mosquito Culex pipiens.
Labbé, Pierrick; Milesi, Pascal; Yébakima, André; Pasteur, Nicole; Weill, Mylène; Lenormand, Thomas
2014-07-01
Gene duplications have long been advocated to contribute to the evolution of new functions. The role of selection in their early spread is more controversial. Unless duplications are favored for a direct benefit of increased expression, they are likely detrimental. In this article, we investigated the case of duplications favored because they combine already functionally divergent alleles. Their gene-dosage/fitness relations are poorly known because selection may operate on both overall expression and duplicates relative dosage. Using the well-documented case of Culex pipiens resistance to insecticides, we compared strains with various ace-1 allele combinations, including two duplicated alleles carrying both susceptible and resistant copies. The overall protein activity was nearly additive, but, surprisingly, fitness correlated better with the relative proportion of susceptible and resistant copies rather than any absolute measure of activity. Gene dosage is thus crucial, duplications stabilizing a "heterozygote" phenotype. It corroborates the view that these were favored because they fix a permanent heterosis, thereby solving the irreducible trade-off between resistance and synaptic transmission. Moreover, we showed that the contrasted successes of the two duplicated alleles in natural populations depend on genetic changes unrelated to ace-1, confirming the probable implication of recessive sublethal mutations linked to structural rearrangements in some duplications. © 2014 The Author(s). Evolution © 2014 The Society for the Study of Evolution.
Modelling the growth of the brown frog (Rana dybowskii)
Du, Xiao-peng; Hu, Zong-fu; Cui, Li-yong
2018-01-01
Well-controlled development leads to uniform body size and a better growth rate; therefore, the ability to determine the growth rate of frogs and their period of sexual maturity is essential for producing healthy, high-quality descendant frogs. To establish a working model that can best predict the growth performance of frogs, the present study examined the growth of one-year-old and two-year-old brown frogs (Rana dybowskii) from metamorphosis to hibernation (18 weeks) and out-hibernation to hibernation (20 weeks) under the same environmental conditions. Brown frog growth was studied and mathematically modelled using various nonlinear, linear, and polynomial functions. The model input values were statistically evaluated using parameters such as the Akaike’s information criterion. The body weight/size ratio (Kwl) and Fulton’s condition factor (K) were used to compare the weight and size of groups of frogs during the growth period. The results showed that the third- and fourth-order polynomial models provided the most consistent predictions of body weight for age 1 and age 2 brown frogs, respectively. Both the Gompertz and third-order polynomial models yielded similarly adequate results for the body size of age 1 brown frogs, while the Janoschek model produced a similarly adequate result for the body size of age 2 brown frogs. The Brody and Janoschek models yielded the highest and lowest estimates of asymptotic weight, respectively, for the body weights of all frogs. The Kwl value of all frogs increased from 0.40 to 3.18. The K value of age 1 frogs decreased from 23.81 to 9.45 in the first four weeks. The K value of age 2 frogs remained close to 10. Graphically, a sigmoidal trend was observed for body weight and body size with increasing age. The results of this study will be useful not only for amphibian research but also for frog farming management strategies and decisions.
The wood frog (Rana sylvatica): a technical conservation assessment
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?|
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
Entomologic studies after a St. Louis encephalitis epidemic in Grand Junction, Colorado.
Tasi, T F; Smith, G C; Ndukwu, M; Jakob, W L; Happ, C M; Kirk, L J; Francy, D B; Lampert, K J
1988-08-01
In 1986, after a St. Louis encephalitis epidemic in Grand Junction, Colorado, in 1985, vector mosquitoes in the city were surveyed to correlate their bionomics and infection rates with the occurrence of human disease. No human cases were reported, but mosquito surveillance disclosed St. Louis encephalitis virus in Culex tarsalis and Culex pipiens pipiens. Mosquitoes were collected with gravid traps designed to attract Cx. p. pipiens and with Centers for Disease Control light traps. Culex p. pipiens was the predominant vector mosquito collected and was captured chiefly in gravid traps. The Culex tarsalis population emerged and expanded approximately one month earlier than did the Cx. p. pipiens population. Consequently, Cx. p. pipiens was the predominant vector species after August. Infection rates throughout the surveillance period (June to September) were severalfold higher in Cx. tarsalis than in Cx. p. pipiens; however, in late summer, diminished numbers of Cx. tarsalis and a persistent population of Cx. p. pipiens resulted in relatively larger numbers of infected Cx. p. pipiens. Thus, the participation of Cx. p. pipiens as a St. Louis encephalitis vector would have been underestimated in previous studies employing light traps alone. These studies provide further evidence that Cx. p. pipiens-associated urban St. Louis encephalitis and rural Cx. tarsalis-associated St. Louis encephalitis cycles may coexist in the West.
Kueneman, Jordan G; Weiss, Sophie; McKenzie, Valerie J
2017-01-01
Global amphibian decline linked to fungal pathogens has galvanized research on applied amphibian conservation. Skin-associated bacterial communities of amphibians have been shown to mediate fungal skin infections and the development of probiotic treatments with antifungal bacteria has become an emergent area of research. While exploring the role of protective bacteria has been a primary focus for amphibian conservation, we aim to expand and study the other microbes present in amphibian skin communities including fungi and other micro-eukaryotes. Here, we characterize skin-associated bacteria and micro-eukaryotic diversity found across life stages of Cascades frog ( Rana cascadae ) and their associated aquatic environments using culture independent 16S and 18S rRNA marker-gene sequencing. Individuals of various life stages of Cascades frogs were sampled from a population located in the Trinity Alps in Northern California during an epidemic of the chytrid fungus, Batrachochytrium dendrobatidis . We filtered the bacterial sequences against a published database of bacteria known to inhibit B. dendrobatidis in co-culture to estimate the proportion of the skin bacterial community that is likely to provide defense against B. dendrobatidis . Tadpoles had a significantly higher proportion of B. dendrobatidis -inhibitory bacterial sequence matches relative to subadult and adult Cascades frogs. We applied a network analysis to examine patterns of correlation between bacterial taxa and B. dendrobatidis , as well as micro-eukaryotic taxa and B. dendrobatidis . Combined with the published database of bacteria known to inhibit B. dendrobatidis , we used the network analysis to identify bacteria that negatively correlated with B. dendrobatidis and thus could be good probiotic candidates in the Cascades frog system.
Impacts of weathered tire debris on the development of Rana sylvatica larvae
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.
Impacts of weathered tire debris on the development of Rana sylvatica larvae.
Camponelli, Kimberly M; Casey, Ryan E; Snodgrass, Joel W; Lev, Steven M; Landa, Edward R
2009-02-01
Highway runoff has the potential to negatively impact receiving systems including stormwater retention ponds where highway particulate matter can accumulate following runoff events. Tire wear particles, which contain about 1% Zn by mass, make up approximately one-third of the vehicle derived particulates in highway runoff and therefore may serve as a stressor to organisms utilizing retention ponds as habitat. In this study, we focused on the potential contribution of tire debris to Zn accumulation by Rana sylvatica larvae and possible lethal or sublethal impacts resulting from exposure to weathered tire debris during development. Eggs and larvae were exposed to aged sediments (containing either ZnCl2 or tire particulate matter, both providing nominal concentrations of 1000 mg Zn kg(-1)) through metamorphosis. Water column Zn was elevated in both the ZnCl2 and tire treatments relative to the control treatment, indicating that aging allowed Zn leaching from tire debris to occur. Tissue Zn was also elevated for the ZnCl2 and tire treatments indicating that Zn in the treatments was available for uptake by the amphibians. Exposure to both ZnCl2 and tire treatments increased the time for larvae to complete metamorphosis in comparison with controls. We also observed that the longer the organisms took to complete metamorphosis, the smaller their mass at metamorphosis. Our results indicate that Zn leached from aged tire debris is bioavailable to developing R. sylvatica larvae and that exposure to tire debris amended sediments can result in measurable physiological outcomes to wood frogs that may influence population dynamics.
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.
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.
1975-01-01
Receiver operating characteristic (ROC) analysis of nerve messages is described. The hypothesis that quantum fluctuations provide the only limit to the ability of frog ganglion cells to signal luminance change information is examined using ROC analysis. In the context of ROC analysis, the quantum fluctuation hypothesis predicts (a) the detectability of a luminance change signal should rise proportionally to the size of the change, (b) detectability should decrease as the square root of background, an implication of which is the deVries-Rose law, and (c) ROC curves should exhibit a shape particular to underlying Poisson distributions. Each of these predictions is confirmed for the responses of dimming ganglion cells to brief luminance decrements at scotopic levels, but none could have been tested using classical nerve message analysis procedures. PMID:172597
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 ...
Orton, Frances; Carr, James A; Handy, Richard D
2006-01-01
Pollution from agrochemicals may be contributing to the global decline in amphibian populations. Environmentally relevant concentrations of nitrate and/or atrazine on anuran development and gonadal differentiation were tested. Four replicates of 20 tadpoles per tank (80/treatment) were exposed from Taylor-Kollros stage 2 to 3 to stage 23 to 34 to either 10 mg/L nitrate, 10 microg/L atrazine, a combined exposure of 10 mg/L nitrate plus 10 microg/L atrazine, or untreated controls. No treatment-dependent effects on weight, snout-vent or hind limb length, or time to forelimb emergence were observed. The proportions of females increased in all treatments compared to the controls, especially in the combined treatment (chi2 = 17.90, df = 6, p = 0.0065, combined = 66.4% female, control = 41% female). The frequency of intersex was low in all treatments. No treatment-related effects on the total number of spermatogenic cells were observed, but the ratio of cell types differed in that testes from animals in the treated groups exhibited more spermatogonia, fewer spermatocytes, and more spermatids than the control (significantly different, Kruskal-Wallis, p < 0.05). Ovaries from animals treated with nitrate or atrazine exhibited larger immature (previtellogenic) and mature (vitellogenic) follicles, but ovaries from the combined treatment had larger immature follicles only. Testicular oocytes were observed in the nitrate-only and atrazine-only treatments, and the control treatment, but not the combined treatment. Overall, this study has demonstrated changes in sex ratios that are more marked in response to combined nitrate/atrazine exposure than with these chemicals alone. Histological evidence suggests that premature maturation of gonad may occur as a result of nitrate and/or atrazine exposure during larval development.
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.
Michaelidis, Basile; Kyriakopoulou-Sklavounou, Pasqualina; Staikou, Alexandra; Papathanasiou, Ioanna; Konstantinou, Kiriaki
2008-12-01
The present work aimed to contribute to the understanding of the adaptation of the glycolytic pathway in tissues of frog Rana ridibunda and land snail species Helix lucorum during seasonal hibernation. Moreover responses of glycolytic enzymes from cold acclimated R. ridibunda and H. lucorum were studied as well. The drop in Po(2) in the blood of hibernated frogs and land snails indicated lower oxygen consumption and a decrease in their metabolic rate. The activities of glycolytic enzymes indicated that hibernation had a differential effect on the glycolyis in the two species studied and also in the tissues of the same species. The activity of l-LDH decreased significantly in the skeletal muscle and heart of hibernated R. ridibunda indicating a low glycolytic potential. Similar biochemical responses were observed in the same tissues during cold acclimation. The continuous increase in the activities of glycolytic enzymes studied, except for HK, might indicate a compensation for the impacts of low temperature on the enzymatic activities. In contrast to R. ridibunda, the activities of the enzymes increased and remained at higher levels than those of the prehibernation controls indicating maintenance of glycolytic potential in the tissues of hibernating land snails.
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.
California red-legged frog (Rana draytonii) movement and habitat use: Implications for conservation
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.
Trujillo, Xóchitl; Ortiz-Mesina, Mónica; Uribe, Tannia; Castro, Elena; Montoya-Pérez, Rocío; Urzúa, Zorayda; Feria-Velasco, Alfredo; Huerta, Miguel
2015-02-01
Previous studies have indicated that vanilloid receptor (VR1) mRNA is expressed in muscle fibers. In this study, we evaluated the functional effects of VR1 activation. We measured caffeine-induced contractions in bundles of the extensor digitorum longus muscle of Rana pipiens. Isometric tension measurements showed that two VR1 agonists, capsaicin (CAP) and N-arachidonoyl-dopamine (NADA), reduced muscle peak tension to 57 ± 4 % and 71 ± 3% of control, respectively. The effect of CAP was partially blocked by a VR1 blocker, capsazepine (CPZ), but the effect of NADA was not changed by CPZ. Because NADA is able to act on cannabinoid receptors, which are also present in muscle fibers, we tested the cannabinoid antagonist AM281. We found that AM281 antagonized both CAP and NADA effects. AM281 alone reduced peak tension to 80 ± 6 % of control. With both antagonists, the CAP effect was completely blocked, and the NADA effect was partially blocked. These results provide pharmacological evidence of the functional presence of the VR1 receptor in fast skeletal muscle fibers of the frog and suggest that capsaicin and NADA reduce tension by activating both cannabinoid and vanilloid receptors.
Defining anural malformations in the context of a developmental problem
Meteyer, C.U.; Cole, Rebecca A.; Converse, K.A.; Docherty, D.E.; Wolcott, M.; Helgen, J.C.; Levey, R.; Eaton-Poole, L.; Burkhart, J.G.
2000-01-01
This paper summarizes terminology and general concepts involved in animal development for the purpose of providing background for the study and understanding of frog malformations. The results of our radiographic investigation of rear limb malformations in Rana pipiens provide evidence that frog malformations are the product of early developmental errors. Although bacteria, parasites and viruses were identified in these metamorphosed frogs, the relevant window to look for the teratogenic effect of these agents is in the early tadpole stage during limb development. As a result, our microbiological findings must be regarded as inconclusive relative to determining their contribution to malformations because we conducted our examinations on metamorphosed frogs not tadpoles. Future studies need to look at teratogenic agents (chemical, microbial, physical or mechanical) that are present in the embryo, tadpole, and their environments at the stages of development that are relevant for the malformation type. The impact of these teratogenic agents then needs to be assessed in appropriate animal models using studies that are designed to mimic field conditions. The results of these laboratory tests should then be analyzed in such a way that will allow comparison with the findings in the wild-caught tadpoles and frogs.
Two-photon excited autofluorescence imaging of freshly isolated frog retinas.
Lu, Rong-Wen; Li, Yi-Chao; Ye, Tong; Strang, Christianne; Keyser, Kent; Curcio, Christine A; Yao, Xin-Cheng
2011-06-01
The purpose of this study was to investigate cellular sources of autofluorescence signals in freshly isolated frog (Rana pipiens) retinas. Equipped with an ultrafast laser, a laser scanning two-photon excitation fluorescence microscope was employed for sub-cellular resolution examination of both sliced and flat-mounted retinas. Two-photon imaging of retinal slices revealed autofluorescence signals over multiple functional layers, including the photoreceptor layer (PRL), outer nuclear layer (ONL), outer plexiform layer (OPL), inner nuclear layer (INL), inner plexiform layer (IPL), and ganglion cell layer (GCL). Using flat-mounted retinas, depth-resolved imaging of individual retinal layers further confirmed multiple sources of autofluorescence signals. Cellular structures were clearly observed at the PRL, ONL, INL, and GCL. At the PRL, the autofluorescence was dominantly recorded from the intracellular compartment of the photoreceptors; while mixed intracellular and extracellular autofluorescence signals were observed at the ONL, INL, and GCL. High resolution autofluorescence imaging clearly revealed mosaic organization of rod and cone photoreceptors; and sub-cellular bright autofluorescence spots, which might relate to connecting cilium, was observed in the cone photoreceptors only. Moreover, single-cone and double-cone outer segments could be directly differentiated.
Descriptive risk assessment of the effects of acidic deposition on Rocky Mountain amphibians
Corn, Paul Stephen; Vertucci, Frank A.
1992-01-01
We evaluated the risk of habitat acidification to the six species of amphibians that occur in the mountains of Colorado and Wyoming. Our evaluation included extrinsic environmental factors (habitat sensitivity and amount of acidic atmospheric deposition) and species-specific intrinsic factors (sensitivity to acid conditions, habitat preferences, and timing of breeding). Only one of 57 surveyed localities had both acid neutralizing capacity μeq/L and sulfate deposition >10 kg/ha/yr, extrinsic conditions with a possible risk of acidification. Amphibian breeding habitats in the Rocky Mountains do not appear to be sufficiently acidic to kill amphibian embryos. Some species breed in high-elevation vernal pools during snowmelt, and an acidic pulse during snowmelt may pose a risk to embryos of these species. However, the acidic pulse, if present, probably occurs before open water appears and before breeding begins. Although inherent variability of amphibian population size may make detection of declines from anthropogenic effects difficult, acidic deposition is unlikely to have caused the observed declines of Bufo boreas and Rana pipiens in Colorado and Wyoming. Amphibians in the Rocky Mountains are not likely to be at risk with acidification inputs at present levels.
Efficient killing of CD22{sup +} tumor cells by a humanized diabody-RNase fusion protein
DOE Office of Scientific and Technical Information (OSTI.GOV)
Krauss, Juergen; Arndt, Michaela A.E.; Vu, Bang K.
2005-06-03
We report on the generation of a dimeric immunoenzyme capable of simultaneously delivering two ribonuclease (RNase) effector domains on one molecule to CD22{sup +} tumor cells. As targeting moiety a diabody derived from the previously humanized scFv SGIII with grafted specificity of the murine anti-CD22 mAb RFB4 was constructed. Further engineering the interface of this construct (V{sub L}36{sub Leu{yields}}{sub Tyr}) resulted in a highly robust bivalent molecule that retained the same high affinity as the murine mAb RFB4 (K{sub D} 0.2 nM). A dimeric immunoenzyme comprising this diabody and Rana pipiens liver ribonuclease I (rapLRI) was generated, expressed as solublemore » protein in bacteria, and purified to homogeneity. The dimeric fusion protein killed several CD22{sup +} tumor cell lines with high efficacy (IC{sub 50} = 3-20 nM) and exhibited 9- to 48-fold stronger cytotoxicity than a monovalent rapLRI-scFv counterpart. Our results demonstrate that engineering of dimeric antibody-ribonuclease fusion proteins can markedly enhance their biological efficacy.« less
Evaluation of use of tiletamine/zolazepam for anesthesia of bullfrogs and leopard frogs.
Letcher, J; Durante, R
1995-07-01
Use of tiletamine hydrochloride and zolazepam hydrochloride (1:1 fixed ratio combination) as an anesthetic agent in 2 anuran species was studied. A dosage of 5 mg/kg of body weight, administered IM, resulted in variable weak tranquilization. Intramuscular administration at dosages of 10 and 20 mg/kg induced variable states of tranquilization or anesthesia in leopard frogs (Rana pipiens) and bullfrogs (R catesbeiana). The dosages of 50 mg/kg induced anesthesia with greater consistency than lower dosages in bullfrogs, but resulted in mortalities. The same dosage was uniformly fatal in leopard frogs. Neither gross nor histologic lesions were identified in the frogs that died. Depth and duration of anesthesia was dosage related. At the 20 and 50 mg/kg dosages, leopard frogs attained a greater depth of anesthesia and remained anesthetized for a significantly greater duration than did bullfrogs; however, at the 5 and 10 mg/kg dosages, bullfrogs developed greater tranquilization for longer periods than did leopard frogs. Results of this study revealed profound intraspecies variation in depth and duration of effect of tiletamine/zolazepam; therefore, the drug does not appear to be a suitable injectable anesthetic in anurans.
Cetin, Huseyin; Dechant, Peter; Yanikoglu, Atila
2007-06-01
Efficacy of tank mixtures of commercial Bacillus thuringiensis subsp. israelensis (Bti) and Bacillus sphaericus (Bs) water-dispersable granule (WDG) formulations was evaluated in septic tanks, against Culex pipiens L. (Diptera: Culicidae) larvae. VectoLex WDG (Bs) + VectoBac WDG (Bti) were evaluated at various ratios from 488 g/ha VectoLex + 250 g/ha VectoBac up to 3,000 g/ha of each. All applications and ratios delivering VectoLex WDG at a rate equal to or greater than 988 g/ha provided more than 90% control for 28 days after treatment. The lowest dose provided this level of control for at least 7 days after treatment, with greater than 80% control after 2 wk. These results suggest that a retreatment interval of 2 wk is recommended with the lowest dose and retreatment intervals of 4 or more wk are recommended with the doses equal to or higher than 988 g/ha VectoLex + 250 g/ha VectoBac.
Experimental transmission of Zika virus by mosquitoes from central Europe.
Heitmann, Anna; Jansen, Stephanie; Lühken, Renke; Leggewie, Mayke; Badusche, Marlis; Pluskota, Björn; Becker, Norbert; Vapalahti, Olli; Schmidt-Chanasit, Jonas; Tannich, Egbert
2017-01-12
Mosquitoes collected in Germany in 2016, including Culex pipiens pipiens biotype pipiens, Culex torrentium and Aedes albopictus, as well as Culex pipiens pipiens biotype molestus (in colony since 2011) were experimentally infected with Zika virus (ZIKV) at 18 °C or 27 °C. None of the Culex taxa showed vector competence for ZIKV. In contrast, Aedes albopictus were susceptible for ZIKV but only at 27 °C, with transmission rates similar to an Aedes aegypti laboratory colony tested in parallel. This article is copyright of The Authors, 2017.
Lind, M I; Johansson, F
2011-12-01
Phenotypic plasticity can be important for local adaptation, because it enables individuals to survive in a novel environment until genetic changes have been accumulated by genetic accommodation. By analysing the relationship between development rate and growth rate, it can be determined whether plasticity in life-history traits is caused by changed physiology or behaviour. We extended this to examine whether plasticity had been aiding local adaptation, by investigating whether the plastic response had been fixed in locally adapted populations. Tadpoles from island populations of Rana temporaria, locally adapted to different pool-drying regimes, were monitored in a common garden. Individual differences in development rate were caused by different foraging efficiency. However, developmental plasticity was physiologically mediated by trading off growth against development rate. Surprisingly, plasticity has not aided local adaptation to time-stressed environments, because local adaptation was not caused by genetic assimilation but on selection on the standing genetic variation in development time. © 2011 The Authors. Journal of Evolutionary Biology © 2011 European Society For Evolutionary Biology.
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.
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.
DOE Office of Scientific and Technical Information (OSTI.GOV)
Amoroso, C.; Lorenz, W.
1959-01-01
Sixty-one frogs (Rana esculenta) were irradiated in urethan anesthesia. The right upper leg was irradiated with a surface dose of 12,000 r. The left upper legs of the same animals were irradiated subsequently with a surface dose of 2 times 6000 r, respectively 3 times 4000 r at intervals of 1, 3, 6, 24, or 72 hours. The single irradiation of 12,000 r caused changes of the pigentation and formation of ulcers after 3 weeks. After 8 weeks 35 out of 61 animals had formation of ulcers. The first tendency for healing was observed after 10 weeks. By the usemore » of divided doses of 3 times 4000 r or 2 tirees 6000 r, with intervals as short as one hour, the effect of the irradiation caused less damage, probably due to increased resistence, which may have developed during the intervals between irradiations. (auth)« less
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
Rheubert, Justin L; Cook, Hanna E; Siegel, Dustin S; Trauth, Stanley E
2017-10-01
Previous studies have revealed variations in the urogenital system morphology of amphibians. Recently, the urogenital system of salamanders was reviewed and terminology was synonymized across taxa. Discrepancies exist in the terminology describing the urogenital system of anurans, which prompted our group to develop a complete, detailed description of the urogenital system in an anuran species and provide nomenclature that is synonymous with those of other amphibian taxa. In Rana catesbeiana, sperm mature within spermatocysts of the seminiferous tubule epithelia and are transported to a series of intratesticular ducts that exit the testes and merge to form vasa efferentia. Vasa efferentia converge into single longitudinal ducts (Bidder's ducts) on the lateral aspects of the kidneys. Branches from the longitudinal ducts merge with genital kidney renal tubules through renal corpuscles. The nephrons travel caudally and empty into the Wöffian ducts. Similar to salamanders, the caudal portion of the kidneys (termed the pelvic kidneys in salamanders) only possesses nephrons involved in urine formation, not sperm transport. Data from the present study provide a detailed description and synonymous nomenclature that can be used to make future comparative analyses between taxa more efficient.
Freitas, Ferdinando B.; Novo, Maria Teresa; Esteves, Aida; de Almeida, A. Paulo G.
2012-01-01
The aim of this study was to evaluate mosquito abundance, species diversity, larval and adult population dynamics in seven lagoons integrated in the wetland coastal system of the Algarve, Portugal, in the summer of 2007, as well as the screening of these for West Nile virus (WNV). WNV has been isolated from mosquitoes in this region, in the summer of 2004, next to the putative area of infection of two linked human WN cases. Adult mosquitoes were collected with CDC traps baited with CO2, and potential breeding sites were surveyed for immature stages. Morphological identification of 1,432 adult mosquitoes and 85 larvae revealed the presence of 10 species: Anopheles atroparvus, Anopheles algeriensis, Coquillettidia richiardii, Culex modestus, Culex pipiens, Culex theileri, Culex univittatus, Culiseta longiareolata, Aedes caspius, and Aedes detritus. Adult mosquito peak densities were recorded in July, contrasting with null larval breeding in the same month in the surveyed biotopes. Most abundant species were C. pipiens (52%), C. theileri (29%), and A. caspius (11%). Lagoon Salgados and Quinta das Salinas, exhibited the highest similarity of culicid fauna, despite being most distant from each other, Female mosquitoes (1,249 specimens) screened by RT-PCR, did not reveal WNV products. However, previous detection of WNV activity in this area, susceptible to re-introductions, demands for continued vigilance. PMID:22347862
Ataie, Abolfazl; Moosa-Kazemi, Seyed Hassan; Vatandoost, Hassan; Yaghoobi-Ershadi, Mohammad Reza; Bakhshi, Hasan; Anjomruz, Mehdi
2015-01-01
Background: Mosquitoes are considered as the vectors of dirofilariasis and some vector borne disease in Iran. The objective of this study was to determine the susceptibility level of the vectors to various insecticides recommended by WHO for any control measures in an endemic area in northwestern Iran. Methods: Mosquito larval and adult collections were carried out using different methods provided by WHO including dipping and hand catch techniques. The susceptibility level was assessed to DDT 4%, malathion 5%, propoxur 0.1%, deltamethrin 0.05% and lambda-cyhalothrin 0.05%. Results: Totally, 749 adults and 5060 larvae of Culicidae mosquitoes were collected comprising seven species of adult and larvae, including: Anopheles claviger, An. maculipennis, An. sacharovi, Culex hortensis, Cx. pipiens, Cx. theileri and Culiseta longiaerolata. Frequency of larvae and adults of An. maculipennis was very low, so susceptibility tests on this species did not performed. Results showed that Cx. theileri, Cs. longiaerolata and Cx. pipiens were resistant to DDT 4%, lambda-cyhalothrin 0.05%, and propoxur 0.1% whereas found tolerant to deltamethrin 0.05% and malathion 5%. The LT50 and LT90 values for five insecticides were calculated. Conclusion: We suggest the same study in different parts of the world to obtain the data due to bionomic and susceptibility status of dirofilariasis vectors. This information will help the health authorities for monitoring and evaluation of control measures. PMID:26114140
Does testis weight decline towards the Subarctic? A case study on the common frog, Rana temporaria
NASA Astrophysics Data System (ADS)
Hettyey, Attila; Laurila, Anssi; Herczeg, Gábor; Jönsson, K. Ingemar; Kovács, Tibor; Merilä, Juha
2005-04-01
Interpopulation comparisons of variation in resource availability and in allocation patterns along altitudinal and latitudinal gradients allow insights into the mechanisms shaping the life history of animals. Patterns of between-population differences in female life history traits have been studied intensively across a wide range of taxa, but similar investigations in males have remained scarce. To study if testis weight—a measure of reproductive investment—varies on a geographical scale in anurans, we focussed on the variation in relative testis weight (RelTW) and asymmetry in 22 populations of the common frog Rana temporaria along a 1,600-km latitudinal transect across the Scandinavian peninsula. We found that RelTW decreased towards the north. Body mass and body length both had independent positive effects on testes mass. We found evidence for directional asymmetry (DA) in testis weight with the right testis being larger than the left. The level of DA in testis weight was not related to latitude, but both body mass and testes mass had independent positive effects on asymmetry. We discuss the northwards decrease in RelTW in terms of a decreased reproductive investment as a possible consequence of harsher environmental conditions, and perhaps also, weaker sexual selection in the north than in the south.
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.
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.
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.
Richter-Boix, Alex; Teplitsky, Céline; Rogell, Björn; Laurila, Anssi
2010-02-01
In ectotherms, variation in life history traits among populations is common and suggests local adaptation. However, geographic variation itself is not a proof for local adaptation, as genetic drift and gene flow may also shape patterns of quantitative variation. We studied local and regional variation in means and phenotypic plasticity of larval life history traits in the common frog Rana temporaria using six populations from central Sweden, breeding in either open-canopy or partially closed-canopy ponds. To separate local adaptation from genetic drift, we compared differentiation in quantitative genetic traits (Q(ST)) obtained from a common garden experiment with differentiation in presumably neutral microsatellite markers (F(ST)). We found that R. temporaria populations differ in means and plasticities of life history traits in different temperatures at local, and in F(ST) at regional scale. Comparisons of differentiation in quantitative traits and in molecular markers suggested that natural selection was responsible for the divergence in growth and development rates as well as in temperature-induced plasticity, indicating local adaptation. However, at low temperature, the role of genetic drift could not be separated from selection. Phenotypes were correlated with forest canopy closure, but not with geographical or genetic distance. These results indicate that local adaptation can evolve in the presence of ongoing gene flow among the populations, and that natural selection is strong in this system.
ANDERSON, SHERI L.; RICHARDS, STEPHANIE L.; TABACHNICK, WALTER J.; SMARTT, CHELSEA T.
2010-01-01
Culex pipiens quinquefasciatus were fed blood containing either 7.0 ± 0.1 logs plaque-forming units (pfu)/ml (high dose) or 5.9 ± 0.1 logs pfu/ml (low dose) of West Nile virus and held at extrinsic incubation temperatures (EIT) of 28°C or 25°C. Approximately 20 mosquitoes per dose were collected after incubation periods (IP) of 4, 6, 8, and 12 days postinfection (dpi). Infection rates were influenced by EIT and virus dose but not by IP. Body titer was significantly higher for mosquitoes fed the high dose and held at 28°C at the later IPs (6, 8, and 12 dpi). However, leg titer was significantly higher for mosquitoes at the later IPs but did not differ between EITs or doses. Because infection rates varied with EIT and dose, there is likely a midgut infection barrier influenced by these factors that is not influenced by IP. Dissemination rates were influenced by all 3 factors consistent with the presence of a midgut escape barrier. Dissemination rate, body titer, and leg titer were dependent on IP, indicating the need to investigate multiple time points in vector competence studies to elucidate critical events in infection and dissemination. PMID:20402358
Anderson, Sheri L; Richards, Stephanie L; Tabachnick, Walter J; Smartt, Chelsea T
2010-03-01
Culex pipiens quinquefasciatus were fed blood containing either 7.0 +/- 0.1 logs plaque-forming units (pfu)/ml (high dose) or 5.9 +/- 0.1 logs pfu/ml (low dose) of West Nile virus and held at extrinsic incubation temperatures (EIT) of 28 degrees C or 25 degrees C. Approximately 20 mosquitoes per dose were collected after incubation periods (IP) of 4, 6, 8, and 12 days postinfection (dpi). Infection rates were influenced by EIT and virus dose but not by IP. Body titer was significantly higher for mosquitoes fed the high dose and held at 28 degrees C at the later IPs (6, 8, and 12 dpi). However, leg titer was significantly higher for mosquitoes at the later IPs but did not differ between EITs or doses. Because infection rates varied with EIT and dose, there is likely a midgut infection barrier influenced by these factors that is not influenced by IP. Dissemination rates were influenced by all 3 factors consistent with the presence of a midgut escape barrier. Dissemination rate, body titer, and leg titer were dependent on IP, indicating the need to investigate multiple time points in vector competence studies to elucidate critical events in infection and dissemination.
Sajid, Mohammed; Ali, Sharique A
2011-08-01
The present study was carried out to determine the effects of lyophilized dried fruit extracts of Piper nigrum and pure piperine on the tadpole melanophores of frog Rana tigerina which offer excellent in vitro opportunities for studying the effects of pharmacological and pharmaceutical agents. The nature of specific cellular receptors present on the neuro-melanophore junction and their involvement in pigmentary responses has been explored. Effects of lyophilized extracts of P. nigrum and pure piperine were studied on the isolated tail melanophores of tadpoles of the frog R. tigerina as per the modified method. The extract of P. nigrum and its active ingredient piperine caused significant melanin dispersal responses leading to darkening of the tail melanophores, which were completely antagonized by atropine and hyoscine. These per se melanin dispersal effects were also found to be markedly potentiated by neostigmine an anticholinesterase agent. It appears that the melanin dispersal effects of the extracts of P. nigrum and pure piperine leading to skin darkening are mediated by cholinergic muscarinic or piperine-like receptors having similar properties.
Independent degeneration of W and Y sex chromosomes in frog Rana rugosa.
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.
Muslu, Hasan; Kurt, Ozgür; Özbilgin, Ahmet
2011-01-01
To identify the mosquito species and the potential mosquito-related infectious diseases in Manisa province, mosquito larvae were collected from aquatic habitats in Manisa between October 2008 and October 2009. Mosquito larvae were collected from the surface water of study sites with a standard larvae collection spoon. The 1st and 2nd stage larvae brought to the laboratory were kept until they become adults, and their species were identified during 3rd or 4th larvae stages. In addition, species identification was made for 3rd or 4th stage larvae as well as pupae in aquatic samples, as well. A total of 8098 larvae samples were collected during the study and Culex (Culex) pipiens and Culex (Neoculex) martini were found to be the predominant species in Manisa. Three Culex [Culex (Culex) pipiens, Culex (Neoculex) martini, Culex (Maillotia) deserticola], two Culiseta [Culiseta (Culiseta) annulata, Culiseta (Allotheobaldia) longiareolata] and one Anopheles [Anopheles (Cellia) superpictus] species were identified. Anopheles superpictus, the vector of malaria; Culex pipiens, Culiseta annulata, Culiseta longiareolata, the vectors of tularemia and arbovirus infections such as West Nile Virus infection, were identified in Manisa province. Conduction of similar larger-scale studies will contribute to the prevention of vector-borne diseases in our region.
DOE Office of Scientific and Technical Information (OSTI.GOV)
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,more » 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.« less
Yu, C.-J.; Debski, E. A.
2008-01-01
Both nicotinic and muscarinic cholinergic receptors are present in the optic tectum. To begin to understand how the activation of these receptors affects visual activity patterns, we have determined the types of physiological responses induced by their activation. Using tectal brain slices from the leopard frog, we found that application of nicotine (100 μM) evoked long-lasting responses in 60% of patch-clamped tectal cells. Thirty percent of these responses consisted of an increase in spontaneous postsynaptic currents (sPSCs) and had both a glutamatergic and GABAergic component as determined by the use of 6-cyano-7-nitroquinoxaline-2,3-dione (50 μM) and bicuculline (25 μM), respectively. Remaining response types consisted of an inward membrane current (16%) and an increase in sPSCs combined with an inward membrane current (14%). All responses could be elicited in the presence of tetrodotoxin (0.5 μM). Muscarinic receptor-mediated responses, induced by carbachol (100 μM) application after nicotinic receptor desensitization, produced responses in 70% of tectal cells. In contrast to responses elicited by nicotine, carbachol-induced responses could be evoked multiple times without significant decrement. Responses consisted of either an outward current (57%), a decrease in sPSCs (5%) or an increase in sPSCs, with (almost 6%) or without (almost 3%) an outward current. The response elicited by carbachol was not predicted by the response of the cell to nicotine. Our results suggest that nicotinic receptors are found predominantly at presynaptic locations in the optic tectum while muscarinic receptors are most often present at postsynaptic sites. We conclude that both of these receptor types could substantially modulate visual activity by changing either the input to tectal neurons or the level of their response to that input. PMID:12676145
Yu, C-J; Debski, E A
2003-01-01
Both nicotinic and muscarinic cholinergic receptors are present in the optic tectum. To begin to understand how the activation of these receptors affects visual activity patterns, we have determined the types of physiological responses induced by their activation. Using tectal brain slices from the leopard frog, we found that application of nicotine (100 microM) evoked long-lasting responses in 60% of patch-clamped tectal cells. Thirty percent of these responses consisted of an increase in spontaneous postsynaptic currents (sPSCs) and had both a glutamatergic and GABAergic component as determined by the use of 6-cyano-7-nitroquinoxaline-2,3-dione (50 microM) and bicuculline (25 microM), respectively. Remaining response types consisted of an inward membrane current (16%) and an increase in sPSCs combined with an inward membrane current (14%). All responses could be elicited in the presence of tetrodotoxin (0.5 microM). Muscarinic receptor-mediated responses, induced by carbachol (100 microM) application after nicotinic receptor desensitization, produced responses in 70% of tectal cells. In contrast to responses elicited by nicotine, carbachol-induced responses could be evoked multiple times without significant decrement. Responses consisted of either an outward current (57%), a decrease in sPSCs (5%) or an increase in sPSCs, with (almost 6%) or without (almost 3%) an outward current. The response elicited by carbachol was not predicted by the response of the cell to nicotine. Our results suggest that nicotinic receptors are found predominantly at presynaptic locations in the optic tectum while muscarinic receptors are most often present at postsynaptic sites. We conclude that both of these receptor types could substantially modulate visual activity by changing either the input to tectal neurons or the level of their response to that input.
de Meillon, Botha; Hayashi, Shigeyo; Sebastian, Anthony
1967-01-01
Although previous workers had found no evidence of resistance to superinfection in vectors of filariasis, it was considered desirable to reinvestigate the subject because of the epidemiological implications, since a mosquito that can incubate to maturity successive broods of filarial larvae will obviously be a more efficient vector than one that cannot. The results obtained indicate that a Culex pipiens fatigans mosquito that picks up an infection early in its life can, by taking subsequent infecting feeds, remain infective for the rest of its life. The movement of mature larvae in the vector and losses of larvae during feeding are of interest since from this information one can estimate the probable number of larvae deposited on the host at each feed and the period of infectivity of the vector. In the present experiment there were, on average, 6.1 larvae per infective mosquito before a second blood-meal; after the meal the figure was 3.6. Thus, 41% of the original infective larvae were lost; not all would be inoculated into the host, as some would die on the surface of his skin. The release of infective larvae through blood-feeding results from a combination of the passive pressure of the engorged blood in the abdomen and the active movement of the larva itself. PMID:5298677
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.
Richards, Stephanie L; Anderson, Sheri L; Lord, Cynthia C; Smartt, Chelsea T; Tabachnick, Walter J
2012-01-01
Culex pipiens quinquefasciatus Say fed blood containing 6.8 +/- 0.3 logs (mean +/- SE) plaque-forming units of West Nile virus (WNV)/ml were maintained at 28 degrees C for incubation periods (IP) of 7, 14, or 21 d. Several attributes of vector competence were determined at each IP using quantitative real-time reverse transcriptase polymerase chain reaction to estimate plaque forming unit equivalents including: infection rate (WNV-positive abdomens), dissemination rate (WNV-positive legs or thoraces), combined dissemination rate (WNV-positive legs and thoraces), transmission rate (WNV-positive saliva), and WNV titers in abdomens, legs, thoraces, and saliva. Each rate increased or was equivalent with increasing IP. Mosquitoes transmitting WNV in saliva also had significantly higher IP-dependent WNV titers in abdomens, legs, and thoraces. Titers of WNV in abdomens were significantly correlated with titers in legs and thoraces, but the degree of association changed with IP. However, titers of abdomens, legs, and thoraces were not correlated with WNV presence or titer in the saliva. The results show that WNV presence or titer in the saliva of infected Cx. p. quinquefasciatus was not directly influenced by processes involved in WNV replication in other tissues. The processes controlling midgut infection and escape are, in part, independent from the infection processes in other tissues. The relationship between infection, dissemination, and transmission varied over time. The infection and replication of WNV in different tissues is likely influenced by different barriers encountered during the extrinsic incubation period. The significance of these observations for understanding vector competence is discussed.
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
Status of the California Red-legged Frog (Rana draytonii) in the State of Baja California, México
Peralta-Garcia, Anny; Hellingsworth, Bradford D.; Richmond, Jonathan Q.; Valdez-Villavicencio, Jorge H.; Ruiz-Campos, Gorgonio; Fisher, Robert N.; Cruz-Hernandez, Pedro; Galina-Tessaro, Patricia
2016-01-01
The California Red-legged Frog (Rana draytonii) is a threatened species in the United States that has undergone population declines, especially in southern California. Due to the lack of information on the status of Mexican populations, we surveyed for the presence of R. draytonii in Baja California and assessed possible threats to population persistence. Our study area extended from the U.S.-Mexican border to the southern end of the distribution of the species in the Sierra San Pedro Mártir. We found R. draytonii at six of 15 historical sites, none at five proxy sites (i.e., alternative sites chosen because the historical record lacked precise locality data), and four at 24 additional sites. The 10 occupied sites are within three watersheds in the Sierra San Pedro Mártir (two sites at Arroyo San Rafael, two sites at Arroyo San Telmo, and six sites at Arroyo Santo Domingo). We did not detect R. draytonii at 60% of historical sites, including the highest elevation site at La Encantada and multiple low-elevation coastal drainages, suggesting the species has declined in Baja California. The threats we noted most frequently were presence of exotic aquatic animal species, water diversion, and cattle grazing. Management of remaining populations and local education is needed to prevent further declines.
Egea-Serrano, Andrés; Hangartner, Sandra; Laurila, Anssi; Räsänen, Katja
2014-04-07
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.
Cailly, Priscilla; Balenghien, Thomas; Ezanno, Pauline; Fontenille, Didier; Toty, Céline; Tran, Annelise
2011-05-06
In this study, carried out in the Camargue region (France), we combined entomological data with geomatic and modelling tools to assess whether the location of breeding sites may explain the spatial distribution of adult mosquitoes. The species studied are important and competent disease vectors in Europe: Culex modestus Ficalbi and Cx. pipiens Linnaeus (West Nile virus), Anopheles atroparvus Van Thiel, a former Plasmodium vector, and An. melanoon Hackett, competent to transmit Plasmodium.Using a logistic regression model, we first evaluated which land cover variables determined the presence of Culex and Anopheles larva. The resulting probability map of larval presence then was used to project the average probability of finding adults in a buffer area. This was compared to the actual number of adults collected, providing a quantitative assessment of adult dispersal ability for each species. The distribution of Cx. modestus and An. melanoon is mainly driven by the repartition of irrigated farm fields and reed beds, their specific breeding habitats. The presence of breeding sites explained the distribution of adults of both species. The buffer size, reflecting the adult dispersal ability, was 700 m for Cx. modestus and 1000 m for An. melanoon. The comparatively stronger correlation observed for Cx. modestus suggested that other factors may affect the distribution of adult An. melanoon. We did not find any association between Cx. pipiens larval presence and the biotope due to the species' ubiquist character. By applying the same method to different species, we highlighted different strengths of association between land cover (irrigated farm fields and reed beds), larval presence and adult population distribution.This paper demonstrates the power of geomatic tools to quantify the spatial organization of mosquito populations, and allows a better understanding of links between landcover, breeding habitats, presence of immature mosquito populations and adult distributions
2011-01-01
Background In this study, carried out in the Camargue region (France), we combined entomological data with geomatic and modelling tools to assess whether the location of breeding sites may explain the spatial distribution of adult mosquitoes. The species studied are important and competent disease vectors in Europe: Culex modestus Ficalbi and Cx. pipiens Linnaeus (West Nile virus), Anopheles atroparvus Van Thiel, a former Plasmodium vector, and An. melanoon Hackett, competent to transmit Plasmodium. Using a logistic regression model, we first evaluated which land cover variables determined the presence of Culex and Anopheles larva. The resulting probability map of larval presence then was used to project the average probability of finding adults in a buffer area. This was compared to the actual number of adults collected, providing a quantitative assessment of adult dispersal ability for each species. Results The distribution of Cx. modestus and An. melanoon is mainly driven by the repartition of irrigated farm fields and reed beds, their specific breeding habitats. The presence of breeding sites explained the distribution of adults of both species. The buffer size, reflecting the adult dispersal ability, was 700 m for Cx. modestus and 1000 m for An. melanoon. The comparatively stronger correlation observed for Cx. modestus suggested that other factors may affect the distribution of adult An. melanoon. We did not find any association between Cx. pipiens larval presence and the biotope due to the species' ubiquist character. Conclusion By applying the same method to different species, we highlighted different strengths of association between land cover (irrigated farm fields and reed beds), larval presence and adult population distribution. This paper demonstrates the power of geomatic tools to quantify the spatial organization of mosquito populations, and allows a better understanding of links between landcover, breeding habitats, presence of immature mosquito
Molecular cloning and characterization of oocyte-specific Pat1a in Rana rugosa frogs.
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. © 2015 Wiley Periodicals, Inc.
Enzymatic regulation of seasonal glycogen cycling in the freeze-tolerant wood frog, Rana sylvatica.
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.
Fitch, Taylor; Villanueva, Gabriela; Quadir, Mohammad M; Sagiraju, Hari K R; Alamgir, Hasanat
2015-07-01
Prevalence and risk factors of PTSD among injured garment workers who survived a major factory collapse. Survivors receiving treatment or rehabilitation care at one year post event were surveyed, which included Post Traumatic Stress Disorder Checklist Specific version. The respondents consisted of 181 people with a mean age of 27.8 years and a majority had less than high school education (91.2%). Multivariable logistic regression found that the odds of having PTSD was higher among married (OR: 3.2 [95% CI: 1.3-8.0]), those who used to work more than 70 hr/week (OR: 2.4 [1.1-5.3]), workers who used to hold higher job positions (OR: 2.6 [1.2-5.6]) or who had a concussion injury (OR: 3.7 [1.4-9.8]). Among the respondents, 83.4% remained unemployed, and only 57.3% (63 people) reported receiving a quarter or less of what they were promised as compensation. Probable PTSD was prevalent among surviving workers of the Rana Plaza building collapse in Bangladesh. © 2015 Wiley Periodicals, Inc.
DOE Office of Scientific and Technical Information (OSTI.GOV)
Sun Wei; Huang Youhua; Zhao Zhe
2006-12-08
The 3{beta}-hydroxysteroid dehydrogenase (3{beta}-HSD) isoenzymes play a key role in cellular steroid hormone synthesis. Here, a 3{beta}-HSD gene homolog was cloned from Rana grylio virus (RGV), a member of family Iridoviridae. RGV 3{beta}-HSD gene has 1068 bp, encoding a 355 aa predicted protein. Transcription analyses showed that RGV 3{beta}-HSD gene was transcribed immediate-early during infection from an initiation site 19 nucleotides upstream of the translation start site. Confocal microscopy revealed that the 3{beta}-HSD-EGFP fusion protein was exclusively colocalized with the mitochondria marker (pDsRed2-Mito) in EPC cells. Upon morphological observation and MTT assay, it was revealed that overexpression of RGV 3{beta}-HSDmore » in EPC cells could apparently suppress RGV-induced cytopathic effect (CPE). The present studies indicate that the RGV immediate-early 3{beta}-HSD gene encodes a mitochondria-localized protein, which has a novel role in suppressing virus-induced CPE. All these suggest that RGV 3{beta}-HSD might be a protein involved in host-virus interaction.« less
Zhang, Luyao; Chen, Xiaoling; Zhang, Ying; Ma, Chengbang; Xi, Xinping; Wang, Lei; Zhou, Mei; Burrows, James F; Chen, Tianbao
2018-03-18
Rana amurensis is important in Chinese medicine as its skin secretions contain abundant bioactive peptides. Here, we have identified the antimicrobial peptide Amurin-2 and three highly-conserved variants, Amurin-2a, Amurin-2b and Amurin-2c through a combination of molecular cloning and MS/MS fragmentation sequencing. Synthetic replicates of these peptides demonstrate potent antimicrobial activity against S. aureus, whilst some have activity against C.albicans and even resistant bacterial MRSA. Furthermore, two Lys-analogues (K 4 -Amurin-2 and K 11 -Amurin-2) were designed to improve the bioactive function and the antimicrobial activity of K 4 -Amurin-2 against E.coli was enhanced distinctly. In addition, the two modified peptides also showed more potent activity against S. aureus, C. albicans and MRSA strains. Meanwhile, these peptides showed inhibitory effect on the cell viability of several cancer cells. As a result, these structural and functional studies of Amurin-2 variants and analogues could provide insights for future antimicrobial peptide design. Copyright © 2018. Published by Elsevier Inc.
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.
Nagano, H; Kiuchi, H; Abe, Y; Shukuya, R
1976-07-01
In the hepatic cytosol fraction of bullfrog, Rana catesbeiana, an alkaline RNase [EC 3.1.4.22] exists in two forms. One is the free form of RNase, which elutes from a carboxymethyl-cellulose column at a concentration of 0.2 M NaC1. The other is a masked or latent form (RNase-RNase inhibitor complex) which is not adsorbed on the carboxymethyl-cellulose column and which can be converted to the free form of RNase by the addition of p-chloromercuribenzoate. Electrophoretically pure RNase was obtained by the following procedure. The unadsorbed fraction of hepatic cytosol on a column of carboxymethyl-cellulose was treated with p-chloromercuribenzoate and then applied to a second carboxymethyl-cellulose column. The molar weight of RNase was determined to be approximately 12,000 by gel filtration and polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. From the results of gel filtration, the molecular weight of the RNase-RNase inhibitor complex was 130,000. The RNase hydrolyzed poly C, poly U, and poly I, but not poly A or poly G. When poly C was used as a substrate, 2',3'-cyclic CMP as an intermediate and 3'-CMP as a final product were identified. The results of amino acid analysis indicated the presence of an unusual component. The general properties of the RNase and the RNase-RNase inhibitor complex are also reported.
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.
Toxic effects of NH4+-N on embryonic development of Bufo gargarizans and Rana chensinensis.
Deng, Hongzhang; Chai, Lihong; Luo, Pingping; Zhou, Meimei; Nover, Daniel; Zhao, Xiaohong
2017-09-01
Although nitrogen fertilizer is commonly used worldwide, little information is currently available about NH 4 + -N toxicity on amphibians. This study determined the acute and chronic toxic effects of NH 4 + -N on two native Chinese amphibian species (Bufo gargarizans and Rana chensinensis), and compared the negative sensitivity of different embryos to NH 4 + -N. Static renewal aqueous exposures were performed using B. gargarizans and R. chensinensis embryos at Gosner stage 2 over 96 h. In terms of 96 h-LC 50 , B. gargarizans and R. chensinensis embryos had significantly different responses to NH 4 + -N, and the latter was more sensitive to NH 4 + -N than the former. In the chronic toxicity test, exposure to 10 mg L -1 NH 4 + -N or higher significantly decreased the hatching rate of embryos in both species. Significant increases in the abnormality rate of embryos at 50 mg L -1 NH 4 + -N or higher were observed and morphological abnormalities were characterized by axial flexures, yolk sac edema, and hyperplasia in both species. Additionally, the total length of embryos decreased in a dose-dependent manner after exposure to NH 4 + -N. The results indicate that NH 4 + -N exposure can increase abnormality and inhibit the hatching and development of embryos in B. gargarizans and R. chensinensis. Copyright © 2017 Elsevier Ltd. All rights reserved.
Nikookar, Seyed Hassan; Fazeli-Dinan, Mahmoud; Azari-Hamidian, Shahyad; Nasab, Seyed Nouraddin Mousavi; Aarabi, Mohsen; Ziapour, Seyyed Payman; Enayati, Ahmadali; Hemingway, Janet
2018-05-04
Mosquitoes are important vectors of human and animal diseases. This study updates current knowledge on fauna, dominance, and distribution of mosquitoes in Mazandaran Province, Northern Iran, to inform disease control effort. Larval collections, using standard dippers or droppers, and adult collections, using total catches, shelter pits, CDC light traps, and human landing catches, were performed monthly in 30 villages across 16 counties, from May to December 2014. Ovitraps, baited with hay infusion as oviposition attractants or stimulants for Aedes (Stegomyia) mosquitoes, were installed in each village and inspected weekly for eggs. Lactophenol and Berlese media were used for preserving and mounting specimens. Overall, 36,024 mosquito specimens (19,840 larvae and 16,184 adults) belonging to 4 genera and 20 species were morphologically identified. The dominance and distribution indices showed that Culex pipiens s.s. was the eudominant species with a constant distribution of larvae (D = 69.07%, C = 100%) and adults (D = 31.86%, C = 100%), followed by Cx tritaeniorhynchus (D = 38.14%, C = 100%) and Anopheles maculipennis s.l. (D = 11.05%, C = 100%) as adults. Aedes vexans was the dominant (7.85%) species, but it had a sporadic (20%) distribution. Culex torrentium and Culiseta morsitans were added as the new species to the checklist of mosquitoes in Mazandaran Province. Due to the potential role, Cx. pipiens s.s. as a vector of various pathogens, further ecological studies are recommended.
Comparative effects of pH and Vision herbicide on two life stages of four anuran amphibian species.
Edginton, Andrea N; Sheridan, Patrick M; Stephenson, Gerald R; Thompson, Dean G; Boermans, Herman J
2004-04-01
Vision, a glyphosate-based herbicide containing a 15% (weight:weight) polyethoxylated tallow amine surfactant blend, and the concurrent factor of pH were tested to determine their interactive effects on early life-stage anurans. Ninety-six-hour laboratory static renewal studies, using the embryonic and larval life stages (Gosner 25) of Rana clamitans, R. pipiens, Bufo americanus, and Xenopus laevis, were performed under a central composite rotatable design. Mortality and the prevalence of malformations were modeled using generalized linear models with a profile deviance approach for obtaining confidence intervals. There was a significant (p < 0.05) interaction of pH with Vision concentration in all eight models, such that the toxicity of Vision was amplified by elevated pH. The surfactant is the major toxic component of Vision and is hypothesized, in this study, to be the source of the pH interaction. Larvae of B. americanus and R. clamitans were 1.5 to 3.8 times more sensitive than their corresponding embryos, whereas X. laevis and R. pipiens larvae were 6.8 to 8.9 times more sensitive. At pH values above 7.5, the Vision concentrations expected to kill 50% of the test larvae in 96-h (96-h lethal concentration [LC50]) were predicted to be below the expected environmental concentration (EEC) as calculated by Canadian regulatory authorities. The EEC value represents a worst-case scenario for aerial Vision application and is calculated assuming an application of the maximum label rate (2.1 kg acid equivalents [a.e.]/ha) into a pond 15 cm in depth. The EEC of 1.4 mg a.e./L (4.5 mg/L Vision) was not exceeded by 96-h LC50 values for the embryo test. The larvae of the four species were comparable in sensitivity. Field studies should be completed using the more sensitive larval life stage to test for Vision toxicity at actual environmental concentrations.
West Nile virus in overwintering Culex mosquitoes, New York City, 2000.
Nasci, R. S.; Savage, H. M.; White, D. J.; Miller, J. R.; Cropp, B. C.; Godsey, M. S.; Kerst, A. J.; Bennett, P.; Gottfried, K.; Lanciotti, R. S.
2001-01-01
After the 1999 West Nile (WN) encephalitis outbreak in New York, 2,300 overwintering adult mosquitoes were tested for WN virus by cell culture and reverse transcriptase-polymerase chain reaction. WN viral RNA and live virus were found in pools of Culex mosquitoes. Persistence in overwintering Cx. pipiens may be important in the maintenance of WN virus in the northeastern United States. PMID:11585542
Effects of octylphenol on the expression of StAR, CYP17 and CYP19 in testis of Rana chensinensis.
Bai, Yao; Li, Xin-Yi; Liu, Zhi-Jun; Zhang, Yu-Hui
2017-04-01
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. Copyright © 2017 Elsevier B.V. All rights reserved.
Schoff, Patrick K; Johnson, Catherine M; Schotthoefer, Anna M; Murphy, Joseph E; Lieske, Camilla; Cole, Rebecca A; Johnson, Lucinda B; Beasley, Val R
2003-07-01
Skeletal malformation rates for several frog species were determined in a set of randomly selected wetlands in the north-central USA over three consecutive years. In 1998, 62 sites yielded 389 metamorphic frogs, nine (2.3%) of which had skeletal or eye malformations. A subset of the original sites was surveyed in the following 2 yr. In 1999, 1,085 metamorphic frogs were collected from 36 sites and 17 (1.6%) had skeletal or eye malformations, while in 2000, examination of 1,131 metamorphs yielded 16 (1.4%) with skeletal or eye malformations. Hindlimb malformations predominated in all three years, but other abnormalities, involving forelimb, eye, and pelvis were also found. Northern leopard frogs (Rana pipiens) constituted the majority of collected metamorphs as well as most of the malformed specimens. However, malformations were also noted in mink frogs (R. septentrionalis), wood frogs (R. sylvatica), and gray tree frogs (Hyla spp.). The malformed specimens were found in clustered sites in all three years but the cluster locations were not the same in any year. The malformation rates reported here are higher than the 0.3% rate determined for metamorphic frogs collected from similar sites in Minnesota in the 1960s, and thus, appear to represent an elevation of an earlier baseline malformation rate.
Woodhams, Douglas C; Voyles, Jamie; Lips, Karen R; Carey, Cynthia; Rollins-Smith, Louise A
2006-04-01
Chytridiomycosis is an emerging infectious disease of amphibians caused by a chytrid fungus, Batrachochytrium dendrobatidis. This panzootic does not equally affect all amphibian species within an assemblage; some populations decline, others persist. Little is known about the factors that affect disease resistance. Differences in behavior, life history, biogeography, or immune function may impact survival. We found that an innate immune defense, antimicrobial skin peptides, varied significantly among species within a rainforest stream amphibian assemblage that has not been exposed to B. dendrobatidis. If exposed, all amphibian species at this central Panamanian site are at risk of population declines. In vitro pathogen growth inhibition by peptides from Panamanian species compared with species with known resistance (Rana pipiens and Xenopus laevis) or susceptibility (Bufo boreas) suggests that of the nine species examined, two species (Centrolene prosoblepon and Phyllomedusa lemur) may demonstrate strong resistance, and the other species will have a higher risk of disease-associated population declines. We found little variation among geographically distinct B. dendrobatidis isolates in sensitivity to an amphibian skin peptide mixture. This supports the hypothesis that B. dendrobatidis is a generalist pathogen and that species possessing an innate immunologic defense at the time of disease emergence are more likely to survive.
Schoff, P.K.; Johnson, C.M.; Schotthoefer, A.M.; Murphy, J.E.; Lieske, C.; Cole, Rebecca A.; Johnson, L.B.; Beasley, V.R.
2003-01-01
Skeletal malformation rates for several frog species were determined in a set of randomly selected wetlands in the north-central USA over three consecutive years. In 1998, 62 sites yielded 389 metamorphic frogs, nine (2.3%) of which had skeletal or eye malformations. A subset of the original sites was surveyed in the following 2 yr. In 1999, 1,085 metamorphic frogs were collected from 36 sites and 17 (1.6%) had skeletal or eye malformations, while in 2000, examination of 1,131 metamorphs yielded 16 (1.4%) with skeletal or eye malformations. Hindlimb malformations predominated in all three years, but other abnormalities, involving forelimb, eye, and pelvis were also found. Northern leopard frogs (Rana pipiens) constituted the majority of collected metamorphs as well as most of the malformed specimens. However, malformations were also noted in mink frogs (R. septentrionalis), wood frogs (R. sylvatica), and gray tree frogs (Hyla spp.). The malformed specimens were found in clustered sites in all three years but the cluster locations were not the same in any year. The malformation rates reported here are higher than the 0.3% rate determined for metamorphic frogs collected from similar sites in Minnesota in the 1960s, and thus, appear to represent an elevation of an earlier baseline malformation rate.
Applying Metabolomics to differentiate amphibian responses ...
Introduction/Objectives/Methods One of the biggest challenges in ecological risk assessment is determining the impact of multiple stressors on individual organisms and populations in ‘real world’ scenarios. Emerging ‘omic technologies, notably, metabolomics, provides an opportunity to address the uncertainties surrounding ecological risk assessment of multiple stressors. The objective of this study was to use a metabolomics biomarker approach to investigate the effect of multiple stressors on amphibian metamorphs. To this end, metamorphs of Rana pipiens (northern leopard frogs) were exposed to the insecticide Carbaryl (0.32 μg/L), a conspecific predator alarm call (Lithobates catesbeianus), Carbaryl and the predator alarm call, and a control with no stressor. In addition to metabolomic fingerprinting, we measured corticosterone levels in each treatment to assess general stress response. We analyzed relative abundances of endogenous metabolites collected in liver tissue with gas chromatography coupled with mass spectrometry. Support vector machine (SVM) methods with recursive feature elimination (RFE) were applied to rank the metabolomic profiles produced. Results/Conclusions SVM-RFE of the acquired metabolomic spectra demonstrated 85-96% classification accuracy among control and all treatment groups when using the top 75 ranked retention time bins. Biochemical fluxes observed in the groups exposed to carbaryl, predation threat, and the combined treatmen
Effect of anoxia on the electroretinogram of three anoxia-tolerant vertebrates.
Stensløkken, Kåre-Olav; Milton, Sarah L; Lutz, Peter L; Sundin, Lena; Renshaw, Gillian M C; Stecyk, Jonathan A W; Nilsson, Göran E
2008-08-01
To survive anoxia, neural ATP levels have to be defended. Reducing electrical activity, which accounts for 50% or more of neural energy consumption, should be beneficial for anoxic survival. The retina is a hypoxia sensitive part of the central nervous system. Here, we quantify the in vivo retinal light response (electroretinogram; ERG) in three vertebrates that exhibit varying degrees of anoxia tolerance: freshwater turtle (Trachemys scripta), epaulette shark (Hemiscyllium ocellatum) and leopard frog (Rana pipiens). A virtually total suppression of ERG in anoxia, probably resulting in functional blindness, has previously been seen in the extremely anoxia-tolerant crucian carp (Carassius carassius). Surprisingly, the equally anoxia-tolerant turtle, which strongly depresses brain and whole-body metabolism during anoxia, exhibited a relatively modest anoxic reduction in ERG: the combined amplitude of turtle ERG waves was reduced by approximately 50% after 2 h. In contrast, the shark b-wave amplitude practically disappeared after 30 min of severe hypoxia, and the frog b-wave was decreased by approximately 75% after 40 min in anoxia. The specific A(1) adenosine receptor antagonist CPT significantly delayed the suppression of turtle ERG, while the hypoxic shark ERG was unaffected by the non-specific adenosine receptor antagonist aminophylline, suggesting adenosinergic involvement in turtle but not in shark.
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.
DOE Office of Scientific and Technical Information (OSTI.GOV)
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 inmore » 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.« less
Perez-Campo, R; López-Torres, M; Paton, D; Sequeros, E; Barja de Quiroga, G
1990-12-01
In the lung of Rana perezi no differences as a function of age have been found for any of the five major antioxidant enzymes, reduced (GSH), oxidized (GSSG) or glutathione ratio (GSSG/GSH), oxygen consumption (VO2) and for in vivo or in vitro stimulated tissue peroxidation. This frog shows a moderate rate of oxygen consumption and a life span substantially longer than that of rats and mice. Chronic (2.5 months) catalase depletion in the lung did not affect survival or any additional antioxidant enzyme, GSH, GSSG or in vivo and in vitro lung peroxidation in any age group. Only the GSSG/GSH ratio and the VO2 were elevated in catalase depleted old but not young frogs. After comparison of these results with those obtained in other animal species by other authors we suggest the possibility that decreases in antioxidant capacity in old age be restricted to species with high basal metabolic rates. Nevertheless, scavenging of oxygen radicals can not be 100% effective in any species. Thus, aging can still be due to the continuous presence of small concentrations of O2 radicals in the tissues throughout the life span in animals with either high or low metabolic rates.
Dynamics of testis-ova in a wild population of Japanese pond frogs, Rana nigromaculata.
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. © 2015 Wiley Periodicals, Inc.
Werblow, A; Flechl, E; Klimpel, S; Zittra, C; Lebl, K; Kieser, K; Laciny, A; Silbermayr, K; Melaun, C; Fuehrer, H-P
2016-03-01
Millions of people die each year as a result of pathogens transmitted by mosquitoes. However, the morphological identification of mosquito species can be difficult even for experts. The identification of morphologically indistinguishable species, such as members of the Anopheles maculipennis complex (Diptera: Culicidae), and possible hybrids, such as Culex pipiens pipiens/Culex pipiens molestus (Diptera: Culicidae), presents a major problem. In addition, the detection and discrimination of newly introduced species can be challenging, particularly to researchers without previous experience. Because of their medical importance, the clear identification of all relevant mosquito species is essential. Using the direct polymerase chain reaction (PCR) method described here, DNA amplification without prior DNA extraction is possible and thus species identification after sequencing can be achieved. Different amounts of tissue (leg, head; larvae or adult) as well as different storage conditions (dry, ethanol, -20 and -80 °C) and storage times were successfully applied and showed positive results after amplification and gel electrophoresis. Overall, 28 different indigenous and non-indigenous mosquito species were analysed using a gene fragment of the COX1 gene for species differentiation and identification by sequencing this 658-bp fragment. Compared with standard PCR, this method is time- and cost-effective and could thus improve existing surveillance and control programmes. © 2015 The Authors. Medical and Veterinary Entomology published by John Wiley & Sons Ltd on behalf of Royal Entomological Society.
Heavy metals in bullfrog (Rana catesbeiana) tadpoles: Effects of depuration before analysis
DOE Office of Scientific and Technical Information (OSTI.GOV)
Burger, J.; Snodgrass, J.
1998-11-01
Although tadpoles may well be excellent organisms to use as bioindicators of heavy metal contamination, the relationship of deposition in the body compared to the tail, and the effect of sediments or other debris in the digestive tract on heavy metal concentrations is unknown. The authors examined the effect of experimental depuration of bullfrog (Rana catesbeiana) tadpoles on heavy metal and selenium concentrations in intact tadpoles, as well as their bodies and tails. They defined depuration in this experiment as allowing defecation as an elimination process for intestinal contents. The authors maintained wild-caught tadpoles in clean water for 0, 24,more » 48, and 72 h to determine the effects of clearing on heavy metal concentrations. They also examined the concentrations of heavy metals in the whole body and digestive tract separately. The authors test the null hypotheses that no differences occur in metals as a function of time in uncontaminated water, and that no differences occur in metal concentrations in the body compared to the tail and to the digestive tract. They rejected these hypotheses based on regression models. Variance in concentrations of chromium (77%) and lead (70%) were explained by part and clearing time; for manganese (80%), mercury (64%), selenium (28%), and cadmium (25%) the variation was explained only by body part; for arsenic (53%), the variation was explained by part, clearing time, and weight of the various parts. For those metals in which clearing time explained part of the variation, metal concentrations in both the body and tail decreased after 24 and 48 h, but increased slightly thereafter. Clearing, however, did not greatly decrease metal concentrations in either the body or tail. These data suggest that for some metals (mercury, manganese, cadmium, selenium), clearing has no effect, and for others the effect is slight. For fresh tadpoles, however, the digestive tract contained significantly higher concentrations of all metals
Clark, D P; Durell, S; Maloy, W L; Zasloff, M
1994-04-08
Antimicrobial peptides comprise a diverse class of molecules used in host defense by plants, insects, and animals. In this study we have isolated a novel antimicrobial peptide from the skin of the bullfrog, Rana catesbeiana. This 20 amino acid peptide, which we have termed Ranalexin, has the amino acid sequence: NH2-Phe-Leu-Gly-Gly-Leu-Ile-Lys-Ile-Val-Pro-Ala-Met-Ile-Cys-Ala-Val-Thr- Lys-Lys - Cys-COOH, and it contains a single intramolecular disulfide bond which forms a heptapeptide ring within the molecule. Structurally, Ranalexin resembles the bacterial antibiotic, polymyxin, which contains a similar heptapeptide ring. We have also cloned the cDNA for Ranalexin from a metamorphic R. catesbeiana tadpole cDNA library. Based on the cDNA sequence, it appears that Ranalexin is initially synthesized as a propeptide with a putative signal sequence and an acidic amino acid-rich region at its amino-terminal end. Interestingly, the putative signal sequence of the Ranalexin cDNA is strikingly similar to the signal sequence of opioid peptide precursors isolated from the skin of the South American frogs Phyllomedusa sauvagei and Phyllomedusa bicolor. Northern blot analysis and in situ hybridization experiments demonstrated that Ranalexin mRNA is first expressed in R. catesbeiana skin at metamorphosis and continues to be expressed into adulthood.
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.
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.
Anti-apoptotic response during anoxia and recovery in a freeze-tolerant wood frog (Rana sylvatica)
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
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.
Göertz, G P; Fros, J J; Miesen, P; Vogels, C B F; van der Bent, M L; Geertsema, C; Koenraadt, C J M; van Rij, R P; van Oers, M M; Pijlman, G P
2016-11-15
Flaviviruses, such as Zika virus, yellow fever virus, dengue virus, and West Nile virus (WNV), are a serious concern for human health. Flaviviruses produce an abundant noncoding subgenomic flavivirus RNA (sfRNA) in infected cells. sfRNA results from stalling of the host 5'-3' exoribonuclease XRN1/Pacman on conserved RNA structures in the 3' untranslated region (UTR) of the viral genomic RNA. sfRNA production is conserved in insect-specific, mosquito-borne, and tick-borne flaviviruses and flaviviruses with no known vector, suggesting a pivotal role for sfRNA in the flavivirus life cycle. Here, we investigated the function of sfRNA during WNV infection of Culex pipiens mosquitoes and evaluated its role in determining vector competence. An sfRNA1-deficient WNV was generated that displayed growth kinetics similar to those of wild-type WNV in both RNA interference (RNAi)-competent and -compromised mosquito cell lines. Small-RNA deep sequencing of WNV-infected mosquitoes indicated an active small interfering RNA (siRNA)-based antiviral response for both the wild-type and sfRNA1-deficient viruses. Additionally, we provide the first evidence that sfRNA is an RNAi substrate in vivo Two reproducible small-RNA hot spots within the 3' UTR/sfRNA of the wild-type virus mapped to RNA stem-loops SL-III and 3' SL, which stick out of the three-dimensional (3D) sfRNA structure model. Importantly, we demonstrate that sfRNA-deficient WNV displays significantly decreased infection and transmission rates in vivo when administered via the blood meal. Finally, we show that transmission and infection rates are not affected by sfRNA after intrathoracic injection, thereby identifying sfRNA as a key driver to overcome the mosquito midgut infection barrier. This is the first report to describe a key biological function of sfRNA for flavivirus infection of the arthropod vector, providing an explanation for the strict conservation of sfRNA production. Understanding the flavivirus transmission
Göertz, G. P.; Fros, J. J.; Miesen, P.; Vogels, C. B. F.; van der Bent, M. L.; Geertsema, C.; Koenraadt, C. J. M.; van Oers, M. M.
2016-01-01
ABSTRACT Flaviviruses, such as Zika virus, yellow fever virus, dengue virus, and West Nile virus (WNV), are a serious concern for human health. Flaviviruses produce an abundant noncoding subgenomic flavivirus RNA (sfRNA) in infected cells. sfRNA results from stalling of the host 5′-3′ exoribonuclease XRN1/Pacman on conserved RNA structures in the 3′ untranslated region (UTR) of the viral genomic RNA. sfRNA production is conserved in insect-specific, mosquito-borne, and tick-borne flaviviruses and flaviviruses with no known vector, suggesting a pivotal role for sfRNA in the flavivirus life cycle. Here, we investigated the function of sfRNA during WNV infection of Culex pipiens mosquitoes and evaluated its role in determining vector competence. An sfRNA1-deficient WNV was generated that displayed growth kinetics similar to those of wild-type WNV in both RNA interference (RNAi)-competent and -compromised mosquito cell lines. Small-RNA deep sequencing of WNV-infected mosquitoes indicated an active small interfering RNA (siRNA)-based antiviral response for both the wild-type and sfRNA1-deficient viruses. Additionally, we provide the first evidence that sfRNA is an RNAi substrate in vivo. Two reproducible small-RNA hot spots within the 3′ UTR/sfRNA of the wild-type virus mapped to RNA stem-loops SL-III and 3′ SL, which stick out of the three-dimensional (3D) sfRNA structure model. Importantly, we demonstrate that sfRNA-deficient WNV displays significantly decreased infection and transmission rates in vivo when administered via the blood meal. Finally, we show that transmission and infection rates are not affected by sfRNA after intrathoracic injection, thereby identifying sfRNA as a key driver to overcome the mosquito midgut infection barrier. This is the first report to describe a key biological function of sfRNA for flavivirus infection of the arthropod vector, providing an explanation for the strict conservation of sfRNA production. IMPORTANCE Understanding
Sanogo, Yibayiri O; Kim, Chang-Hyun; Lampman, Richard; Novak, Robert J
2007-07-01
In North America, West Nile and St. Louis encephalitis viruses have been detected in a wide range of vector species, but the majority of isolations continue to be from pools of mixed mosquitoes in the Culex subgenus Culex. Unfortunately, the morphologic identification of these important disease vectors is often difficult, particularly in regions of sympatry. We developed a sensitive real-time TaqMan polymerase chain reaction assay that allows reliable identification of Culex mosquitoes including Culex pipiens pipiens, Cx. p. quinquefasciatus, Cx. restuans, Cx. salinarius, Cx. nigripalpus, and Cx. tarsalis. Primers and fluorogenic probes specific to each species were designed based on sequences of the acetylcholinesterase gene (Ace2). Both immature and adult mosquitoes were successfully identified as individuals and as mixed species pools. This identification technique provides the basis for a rapid, sensitive, and high-throughput method for expounding the species-specific contribution of vectors to various phases of arbovirus transmission.
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.
Wojnarowicz, Pola; Ogunlaja, Olumuyiwa O; Xia, Chen; Parker, Wayne J; Helbing, Caren C
2013-12-03
Improved endocrine disrupting compound (EDC) removal is desirable in municipal wastewater treatment plants (MWWTPs) although increased removal does not always translate into reduced biological activity. Suitable methods for determining reduction in biological activity of effluents are needed. In order to determine which MWWTPs are the most effective at removing EDC activities, we operated three configurations of pilot sized biological reactors (conventional activated sludge, CAS; nitrifying activated sludge, NAS; and biological nutrient removal, BNR) receiving the same influent under simulated winter and summer conditions. As frogs are model organisms for the study of thyroid hormone (TH) action, we used the North American species Rana catesbeiana in a cultured tadpole tailfin (C-fin) assay to compare the effluents. TH-responsive (thyroid hormone receptors alpha (thra) and beta (thrb)) and stress-responsive (superoxide dismutase, catalase, and heat shock protein 30) mRNA transcript levels were examined. Effluents infrequently perturbed stress-responsive transcript abundance but thra/thrb levels were significantly altered. In winter conditions, CAS caused frequent TH perturbations while BNR caused none. In summer conditions, however, BNR caused substantial TH perturbations while CAS caused few. Our findings contrast other studies of seasonal variations of EDC removal and accentuate the importance of utilizing appropriate biological readouts for assessing EDC activities.
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.
2015-01-01
Culex pipiens, an invasive mosquito and vector of West Nile virus in the US, has two morphologically indistinguishable forms that differ dramatically in behavior and physiology. Cx. pipiens form pipiens is primarily a bird-feeding temperate mosquito, while the sub-tropical Cx. pipiens form molestus thrives in sewers and feeds on mammals. Because the feral form can diapause during the cold winters but the domestic form cannot, the two Cx. pipiens forms are allopatric in northern Europe and, although viable, hybrids are rare. Cx. pipiens form molestus has spread across all inhabited continents and hybrids of the two forms are common in the US. Here we elucidate the genes and gene families with the greatest divergence rates between these phenotypically diverged mosquito populations, and discuss them in light of their potential biological and ecological effects. After generating and assembling novel transcriptome data for each population, we performed pairwise tests for nonsynonymous divergence (Ka) of homologous coding sequences and examined gene ontology terms that were statistically over-represented in those sequences with the greatest divergence rates. We identified genes involved in digestion (serine endopeptidases), innate immunity (fibrinogens and α-macroglobulins), hemostasis (D7 salivary proteins), olfaction (odorant binding proteins) and chitin binding (peritrophic matrix proteins). By examining molecular divergence between closely related yet phenotypically divergent forms of the same species, our results provide insights into the identity of rapidly-evolving genes between incipient species. Additionally, we found that families of signal transducers, ATP synthases and transcription regulators remained identical at the amino acid level, thus constituting conserved components of the Cx. pipiens proteome. We provide a reference with which to gauge the divergence reported in this analysis by performing a comparison of transcriptome sequences from conspecific
Rajjo, I M; Vigna, S R; Crim, J W
1988-04-01
The distribution of the intestinal hormone, cholecystokinin (CCK), was studied in the gastrointestinal tract of a holostean fish, the bowfin (Amia calva), and compared to a teleostean fish, the bluegill (Lepomis macrochirus), and an amphibian, the bullfrog (Rana catesbeiana), using an antiserum specific for the carboxyl terminal tetrapeptide of CCK in an unlabeled biotin-avidin immunoperoxidase procedure. In the bowfin CCK immunostained cells were detected only in the anterior and mid-intestine; the stomach and the rest of the gastrointestinal tract were negative. Immunoreactive cells were open in appearance and were scattered along the intestinal mucosal epithelium, with cells in mid-intestine relatively more abundant than in anterior intestine. These relative distributions were confirmed by radioimmunoassay of tissue extracts. Additional immunostained cells of uncertain function were detected in the lamina propria of the intestine. In bluegill gut immunoreactive cells were observed in the anterior and mid-intestine and in the pyloric caeca, where cells were clustered near the intestinal opening. Immunoreactive cells occurred relatively uniformly along the anterior and mid-intestine. Bullfrog CCK-containing cells were detected both in the antral stomach and in the duodenum. CCK in gut tissues likely originated in the intestine. The redistribution of CCK cells toward the anterior part of the intestine during evolution coincides with the development of a compact pancreas in higher classes of vertebrates. Such a redistribution constitutes an adaptive placement of endocrine cells for signaling during the intestinal phase of digestion.
Lance, Stacey L; Erickson, Matthew R; Flynn, R Wesley; Mills, Gary L; Tuberville, Tracey D; Scott, David E
2012-07-01
Exposure to environmental contaminants contributes to the global decline of amphibian populations. The impacts of organic contaminants on amphibians are well documented. However, substantially less is known concerning the potential effects of metals on amphibian populations. Copper (Cu) is an essential element, but it can be toxic at concentrations only slightly higher than the normal physiological range. The present study examines the effects of chronic Cu exposure on embryos and larvae of southern leopard frogs, Lithobates (Rana) sphenocephalus. Groups of eggs from multiple clutches were collected from two wetlands and exposed to a range of Cu concentrations (0-150 µg/L) until they reached the free-swimming stage, and then individual larvae were reared to metamorphosis. Higher Cu concentrations significantly reduced embryo survival to the free-swimming stage but did not further reduce survival to metamorphosis. Larval period was affected by Cu treatment, but the clutch from which larvae originated (i.e., parentage) explained a higher proportion of the variation. Embryo survival to hatching varied significantly among clutches, ranging from 42.9 to 79.2%. Measurable levels of Cu were found in larvae with body burdens up to 595 µg Cu/g dry mass in the 100 µg/L treatment, and larval Cu body burdens were higher than in metamorphs. The present study also demonstrated that higher initial egg density ameliorated embryo mortality at higher Cu levels and should be accounted for in future studies. Copyright © 2012 SETAC.
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. Copyright © 2011 Elsevier GmbH. All rights reserved.
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.
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.
DOE Office of Scientific and Technical Information (OSTI.GOV)
Peterson, J.D.; Peterson, V.A.; Mendonca, M.T.
Coal combustion residues (CCRs) are documented to negatively impact oral morphology, growth, and development in larval amphibians. It is currently unclear what physiological mechanisms may mediate these effects. Corticosterone, a glucocorticoid hormone, is a likely mediator because when administered exogenously it, like CCRs, also negatively influences oral morphology, growth, and development in larval amphibians. In an attempt to identify if corticosterone mediates these effects, we raised larval Southern Leopard Frogs, Rana sphenocephala, on either sand or CCR substrate and documented effects of sediment type on whole body corticosterone, oral morphology, and time to and mass at key metamorphic stages. Coalmore » combustion residue treated tadpoles contained significantly more corticosterone than controls throughout metamorphosis. However, significantly more oral abnormalities occurred early in metamorphosis when differences in corticosterone levels between treatments were minimal. Overall, CCR-treated tadpoles took significantly more time to transition between key stages and gained less mass between stages than controls, but these differences between treatments decreased during later stages when corticosterone differences between treatments were greatest. Our results suggest endogenous increase in corticosterone content and its influence on oral morphology, growth and development is more complex than previously thought.« less
Sound and vibration sensitivity of VIIIth nerve fibers in the grassfrog, Rana temporaria.
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
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.
Schumann, W
1974-01-01
By applying immunologic, electrophoretic and enzymatic methods, extracts of different raft types of Culex pipiens were analysed. Rafts of the crosses Pa x Pa and Ha x Ha contained four common antigens, while unfertilized rafts of Pa and Ha (no antisera were prepared against them) and rafts of the crosses Og x Og, Og x Pa, and Pa x Og shared three common antigens with the remaining raft extracts. Disk-electrophoresis of raft extracts in acrylamide gel resulted in different electropherograms. Ten protein bands were common to all these raft types. The unfertilized rafts of Pa and Ha yielded three more protein bands, the crosses Pa x Ha and Ha x Pa one more, the crosses Og x Og and Pa x Og three more, and Og x Pa two more. Many enzymes were demonstrated in the raft extracts after they were separated in acrylamide gel and incubated with the corresponding substrate solutions. All the raft types possessed one enzyme type for glutaminate-, lactate-, glucose-6-phosphate-dehydrogenase and catalase. Malate-dehydrogenase and leucine aminopeptidase occurred in each raft type as two isoenzymes. Alkaline phosphatase was observed as a single enzyme, but was lacking in rafts of the crosses Pa X Pa and Ha X Ha. While rafts of the crosses Og x Og and Og x Pa possessed two acid phosphatases, three could be demonstrated for the remaining raft types. Up to eight esterases appeared; rafts of the crosses Og x Og and Og x Pa possessed seven such activities. The results obtained by the Ouchterlony test, disk-electrophoresis and the histochemical enzyme tests are discussed in context and checked according to the phenomenon of incompatibility.
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
Glahn, David; Nuccitelli, Richard
2003-04-01
Voltage-clamped mature, jelly-intact Xenopus eggs were used to carefully examine the ionic currents crossing the plasma membrane before, during, and after fertilization. The bulk of the fertilization current was transient, of large amplitude, and reversed at the predicted Cl- reversal potential. However, the large amplitude fertilization current was preceded by a small, step-like increase in holding current. This small increase in holding current is referred to in this paper as Ion to acknowledge its qualitative similarity to the Ion current previously described in the sea urchin. It was observed in both fertilized and artificially activated eggs, and was found to be unaffected by 10 mm tetra-ethyl ammonium (TEA), a concentration found to block K+ currents in Rana pipiens. Current-voltage relationships are presented for the large fertilization potential, and show that the fertilization currents have a marked outward rectification and are voltage sensitive. These properties are in contrast to the total lack of rectification and slight voltage sensitivity seen before or after the fertilization currents. The time required for sperm to fertilize the egg was found to be voltage dependent with a relatively more depolarized voltage requiring a longer time for fertilization to occur. The percentage of eggs blocked with varying potential levels was determined and this information was fitted to a modified Boltzmann equation having a midpoint of -9 mV.
Schotthoefer, A.M.; Cole, Rebecca A.; Beasley, V.R.
2003-01-01
The effect of echinostome infections on the survival of Rana pipiens tadpoles was examined in relation to developmental stage of tadpoles. Individual tadpoles of Gosner stages 25, 27, 32–33, and 37–39 were exposed to 1 of 4 levels of cercariae (0, 20, 50, or 100). Only tadpoles at stage 25, the earliest stage infected, died within a 5-day experimental period. This stage-specific mortality rate could be explained, in part, by the stage-specific location of encystment of cercariae, which was documented in a separate experiment. In accordance with kidney development, cercariae predominately encysted in the pronephroi during early stages of tadpole development (stages 25 through 31–32) and only in the mesonephroi and associated ducts at later stages (stages 37 through 46). As the mesonephros develops, renal capacity presumably increases. Thus, tadpoles died only when metacercariae concentrated in the functional portion of the kidney with the most limited renal capacity. As tadpoles aged, they also became less susceptible to infections. On average, 69.5% of cercariae that were exposed to stage 25–26 tadpoles successfully encysted, compared with only 8.4% of cercariae exposed to stage 37–38 tadpoles. Exposures of metamorphic frogs (poststage 46) to cercariae revealed that these individuals can become infected with echinostomes. Collectively, our data highlight the host stage–dependent dynamics of tadpole–echinostome interactions.
Agrochemicals increase trematode infections in a declining amphibian species.
Rohr, Jason R; Schotthoefer, Anna M; Raffel, Thomas R; Carrick, Hunter J; Halstead, Neal; Hoverman, Jason T; Johnson, Catherine M; Johnson, Lucinda B; Lieske, Camilla; Piwoni, Marvin D; Schoff, Patrick K; Beasley, Val R
2008-10-30
Global amphibian declines have often been attributed to disease, but ignorance of the relative importance and mode of action of potential drivers of infection has made it difficult to develop effective remediation. In a field study, here we show that the widely used herbicide, atrazine, was the best predictor (out of more than 240 plausible candidates) of the abundance of larval trematodes (parasitic flatworms) in the declining northern leopard frog Rana pipiens. The effects of atrazine were consistent across trematode taxa. The combination of atrazine and phosphate--principal agrochemicals in global corn and sorghum production--accounted for 74% of the variation in the abundance of these often debilitating larval trematodes (atrazine alone accounted for 51%). Analysis of field data supported a causal mechanism whereby both agrochemicals increase exposure and susceptibility to larval trematodes by augmenting snail intermediate hosts and suppressing amphibian immunity. A mesocosm experiment demonstrated that, relative to control tanks, atrazine tanks had immunosuppressed tadpoles, had significantly more attached algae and snails, and had tadpoles with elevated trematode loads, further supporting a causal relationship between atrazine and elevated trematode infections in amphibians. These results raise concerns about the role of atrazine and phosphate in amphibian declines, and illustrate the value of quantifying the relative importance of several possible drivers of disease risk while determining the mechanisms by which they facilitate disease emergence.
Effects of cadmium-enriched sediment on fish and amphibian embryo-larval stages
DOE Office of Scientific and Technical Information (OSTI.GOV)
Francis, P.C.; Birge, W.J.; Black, J.A.
1984-08-01
Aquatic toxicity tests were conducted to evaluate the effects of cadmium-enriched sediment on embryo-larval stages of the goldfish (Carassius auratus), leopard frog (Rana pipiens), and largemouth bass (Micropterus salmoides). Natural stream sediment was collected and enriched with cadmium to nominal concentrations of 1.0, 10.0, 100, and 1000 mg/kg. Enriched sediments were placed in Pyrex dishes and covered with 350 ml of reconstituted water. Fertilized eggs were placed in the dishes and maintained through 4 days posthatching, giving a total exposure time of 6 to 7 days. For all tests the cadmium concentrations ranged from 1.1 to 76.5 micrograms/liter in watermore » above sediments containing 1 to 1000 mg Cd/kg, respectively. Although low frequencies of mortality were observed in all tests, goldfish, leopard frog, and bass exposed to sediments enriched to 1000 mg Cd/kg accumulated 4.61, 12.55, and 60.0 micrograms Cd/g, respectively. No significant correlations were found between mortality of the goldfish and leopard frog and the cadmium concentrations in either water or sediment. However, all three species showed strong correlations between cadmium concentrations in water and tissue, sediment and tissue, and water and sediment. Tissue cadmium concentrations were related to the length of time test organisms were in direct contact with cadmium-enriched sediment.« less
Ecological Effects of Exotic and Native Aquatic Vegetation
2009-08-01
surveys, with American bullfrogs (Rana catesbiena) the most frequently observed. Red-eared sliders ( Trachemys scripta elegans ) were the only...Ernst et al. 1994), with the bulk of their diet consisting of aquatic vegetation. Research has shown that red-eared sliders (T. scripta elegans ) feed...macrophytes for cover, and in some cases for food. Semi-aquatic turtles of the genus Trachemys , for instance, are primarily herbivorous as adults
Origin of sex chromosomes in six groups of Rana rugosa frogs inferred from a sex-linked DNA marker.
Oike, Akira; Watanabe, Koichiro; Min, Mi-Sook; Tojo, Koji; Kumagai, Masahide; Kimoto, Yuya; Yamashiro, Tadashi; Matsuo, Takanori; Kodama, Maho; Nakamura, Yoriko; Notsu, Masaru; Tochimoto, Takeyoshi; Fujita, Hiroyuki; Ota, Maki; Ito, Etsuro; Yasumasu, Shigeki; Nakamura, Masahisa
2017-08-01
Each vertebrate species, as a general rule, has either the XX/XY or ZZ/ZW chromosomes by which sex is determined. However, the Japanese Rana (R.) rugosa frog is an exception, possessing both sex-determining combinations within one species, varying with region of origin. We collected R. rugosa frogs from 104 sites around Japan and South Korea and determined the nucleotide sequences of the mitochondrial 12S ribosomal RNA gene. Based on the sequences, R. rugosa frogs were divided into four groups from Japan and one from South Korea. The ZZ/ZW type is reportedly derived from the XX/XY type, although recently a new ZZ/ZW type of R. rugosa was reported. However, it still remains unclear from where the sex chromosomes in the five groups of this species were derived. In this study, we successfully isolated a sex-linked DNA maker and used it to classify R. rugosa frogs into several groupings. From the DNA marker as well as from nucleotide analysis of the promoter region of the androgen receptor (AR) gene, we identified another female heterogametic group, designated, West-Central. The sex chromosomes in the West-Central originated from the West and Central groups. The results indicate that a sex-linked DNA marker is a verifiable tool to determine the origin of the sex chromosomes in R. rugosa frogs in which the sex-determining system has changed, during two independent events, from the male to female heterogamety. © 2017 Wiley Periodicals, Inc.
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
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. Copyright © 2016 Elsevier Ltd. All rights reserved.
Richards, Stephanie L; Mores, Christopher N; Lord, Cynthia C; Tabachnick, Walter J
2007-01-01
Culex pipiens quinquefasciatus Say mosquitoes from a laboratory colony were exposed to artificial blood meals containing West Nile virus (WNV) and held at incubation temperatures approximating average daily temperatures that occur during Florida arboviral periods. Mosquitoes fed blood meals containing 6.2 logs plaque-forming units (pfu) WNV/mL and held at 25 degrees C, 28 degrees C, or 30 degrees C for 13 days exhibited significantly different rates of infection (30%, 52%, 93%) and dissemination (33%, 22%, 81%) across temperatures. In a separate experiment, Cx. p. quinquefasciatus mosquitoes were provided artificial blood meals with graded doses of WNV from 3.7 to 5.8 logs pfu/mL and maintained at 28 degrees C for 13 days. Rates of infection increased as a function of virus dose, but neither body titers nor dissemination rates were significantly different for mosquitoes that were infected by ingesting different amounts of WNV. Our findings indicate that efficiency of WNV infection and dissemination, and thereby transmission, in Cx. p. quinquefasciatus populations similar to our tested colony may also be diminished when fed blood meals containing less than 5.8 logs pfu WNV/mL and when environmental temperature falls below 30 degrees C. The relationship between the infection rate and dissemination rate changed at different temperatures. This relationship is likely complex and dependent on diverse interactions between factors such as incubation temperature and viremia, which should also be assessed for field populations.
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
Use of femur bone density to segregate wild from farmed Dybowski's frog (Rana dybowskii).
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. Copyright © 2010 Elsevier Ireland Ltd. All rights reserved.
The effects of purine compounds on the isolated aorta of the frog Rana temporaria.
Knight, G. E.; Burnstock, G.
1996-01-01
1. In the isolated aorta of the frog, Rana temporaria, adenosine concentration-dependently, endothelium-independently relaxed adrenaline pre-constricted vessels. None of the adenosine analogues including D-5'-(N-ethylcarboxamide) adenosine (NECA), R- and S-N6-(2-phenylisopropyl) adenosine (R-and S-PIA) and 2-chloroadenosine (2-CA), or the more selective A1, A2 and A3 agonists cyclopentyladenosine (CPA), CGS 21680 and N6-(3-iodobenzyl) adenosine-5'-N-methylcarboxamide (IB-MECA) respectively, had any effect. 2. The non-selective adenosine antagonist, 8-p-sulphophenyl-theophylline (8-pSPT; 30 microM) failed to inhibit adenosine relaxations, as did NG-nitro-L-arginine methyl ester (L-NAME; 0.1 mM) and indomethacin (30 microM). 3. Adenosine 5'-triphosphate (ATP), alpha, beta-methylene ATP (alpha, beta-MeATP), beta, gamma-methylene ATP (beta, gamma-MeATP), 2-methylthio ATP (2-MeSATP) and uridine 5'-triphosphate (UTP) all concentration-dependently contracted the frog aorta. ATP and alpha, beta-MeATP were equipotent and more potent than UTP and beta, gamma-MeATP; 2-MeSATP had little activity. 4. The P2-purinoceptor antagonist, suramin (0.1 mM) inhibited contractions to alpha, beta-MeATP but not to ATP. Pyridoxalphosphate-6-azophenyl-2',4'-disulphonic acid (PPADS; 30 microM) also inhibited contractions to alpha, beta-MeATP but not to ATP. Contractions to ATP were, however, inhibited by indomethacin (30 microM). 5. In conclusion, in the frog aorta there appears to be a novel subclass of P1-purinoceptor mediating vasodilatation, although like the A3 subclass it is not blocked by methylxanthines; a P2-purinoceptor mediates vasconstriction which resembles a P2x subtype, based on the agonist potency of alpha, beta-MeATP being more potent than 2-MeSATP (UTP has moderate activity) and PPADS is an effective antagonist. There is no evidence for the presence of a P2y-purinoceptor, mediating vasodilatation, in this preparation. PMID:8851504
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.
Dowling, Zara; Ladeau, Shannon L; Armbruster, Peter; Biehler, Dawn; Leisnham, Paul T
2013-07-01
Mosquito populations are largely regulated by processes occurring at the larval stage. We sampled mosquito larval microhabitats (mostly water-holding containers) in six neighborhoods in the Washington, DC, area that varied in socioeconomic status (SES) and housing structure (row houses vs. stand-alone houses) to test associations among these neighborhood characteristics, microhabitat abundance and parameters, and mosquito occurrence and densities. Thirty-four percent (33.9%) of sampled microhabitats contained mosquito larvae, and 93.1% of larvae were Aedes albopictus Skuse or Culex pipiens L. Five specific container types (drains, corrugated flexible drainpipes, planters, garbage cans, and buckets) accounted for the majority of water-holding (56.0%) and mosquito-positive (50.6%) microhabitats sampled. We found no associations between SES or housing structure with total microhabitat abundance per yard, mosquito occurrence or mosquito densities per microhabitat. In contrast, container purpose varied with SES, with low SES neighborhoods having greater numbers of disused containers and lower numbers of functional containers than low and medium SES neighborhoods. Ae. albopictus were 83% more abundant in disused containers, whereas Cx. pipiens were more abundant in structural and functional containers, possibly owing to species-specific oviposition and development related to water quality. Ae. albopictus densities increased over the summer, whereas Cx. pipiens densities remained constant. Ae. albopictus is usually the dominant pest in urban areas in the eastern United States; therefore, integrated mosquito management programs should incorporate the elimination of disused containers to reduce its infestation and adult production, especially in low SES neighborhoods where they occur most frequently.
Ferguson, Laura V.; Kirk Hillier, N.; Smith, Todd G.
2012-01-01
Hepatozoon species are heteroxenous parasites that commonly infect the blood of vertebrates and various organs of arthropods. Despite their ubiquity, little is known about how these parasites affect host phenotype, including whether or not these parasites induce changes in hosts to increase transmission success. The objectives of this research were to investigate influences of the frog blood parasite Hepatozoon clamatae and the snake blood parasite Hepatozoon sipedon on host-seeking and host-choice behaviour of the mosquitoes Culex territans and Culex pipiens, respectively. During development of H. sipedon in C. pipiens, significantly fewer infected mosquitoes fed on uninfected snakes compared to uninfected mosquitoes. When H. sipedon was mature in C. pipiens, the number of infected and uninfected C. pipiens that fed on snakes was not significantly different. Higher numbers of mosquitoes fed on naturally infected snakes and frogs compared to laboratory-reared, uninfected control animals. However, experiments using only laboratory-raised frogs revealed that infection did not significantly affect host choice by C. territans. Behaviour of C. pipiens in the presence of H. sipedon may increase transmission success of the parasite and provide the first evidence of phenotypic changes in the invertebrate host of Hepatozoon parasites. PMID:24533317
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
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.
Qureshi, Irfan Zia; Kashif, Zeshan; Hashmi, Muhammad Zaffar; Su, Xiaomei; Malik, Riffat Naseem; Ullah, Kalim; Hu, Jinxing; Dawood, Muhammad
2015-09-01
In the present study, we investigated the concentrations of Ni, Fe, Pb, Cu, Co, Zn, Cd, Mn, and Cr in selected body tissues (liver, stomach, kidney, heart, lungs, and skeletal muscles) of two frog species: Rana tigrina and Euphlyctis cyanophlyctis captured from industrial wastewater of Sialkot city known worldwide for its tanning industry. The both frog species had darker appearance, distinctively different wet body weight, and snout-vent length. The results revealed that the heavy metal concentrations were high in the samples collected from industrial sites as compared to non-industrial sites. The different tissues of R. tigrina and E. cyanophlyctis exhibited little significant differences from two sites. The concentrations of heavy metals were more in tissues of R. tigrina as compared to E. cyanophlyctis. Mean concentration of Cd, Fe, Ni, Mn, Cu, and Cr was comparatively greater in R. tigrina, whereas Pb and Co were higher in E. cyanophlyctis. The concentration of Cu and Cd in the liver and kidney were relatively more in both species as compared to other organs. Further, the results indicated that frogs collected from industrial sites showed decreased body length and weight, and greater metal accumulation. The results will help the authorities for the conservation of these frog species which are under the influence of heavy metal contamination.
Corn, Paul Stephen; Stolzenburg, William; Bury, R. Bruce
1989-01-01
Acid deposition may be detrimental or stressful to native populations of wildlife. Because many species of amphibians breed in shallow ponds created by spring rains or melting snow, they may be particularly vulnerable to the effects of acidification. From 1986 to 1988, we surveyed 105 locations in the central Rocky Mountains where amphibians had been recorded previously, and we found that two species of amphibians had experiences major losses. We found the northern leopard frog (Rana pipiens) at only 4 of 33 (12%) historically known localities, and the boreal toad (Bufo boreas) was present at 10 of 59 (17%) known localities. Three other species have not suffered region-wide declines. Tiger salamanders (Ambystoma tigrinum) and wood frogs (Rana sylvatica) were present at 45% and 69% of known localities respectively, and were observed at several localities were they had not been recorded previously. Chorus frogs (Pseudacris triseriata) suffered a catastrophic decline in population size in one population monitored since 1961, but regionally, this species was observed in 36 of 56 (64%) known localities and in another 19 localities where there were no previous records. Complete water chemistry was recorded for 41 localities, and pH was measured at 110 sites in total. Acid neutralizing capacity, pH, specific conductivity, and cation concentrations were negatively correlated with elevation. However, in mountain ponds and lakes, pH was rarely less than 6.0 during the amphibian breeding season. We tested the tolerance of embryos of the four species of frogs to low pH. The LC50 pH was 4.8 for chorus frogs, 4.4-4.7 for leopard frogs, 4.4-4.5 for boreal toads, and 4.2-4.3 for wood frogs. Survival of wood frog embryos declined when exposed to aluminum concentrations of 100 µg/L or greater, but boreal toad embryos survived exposure to aluminum concentrations of 400 µg/L. Acid deposition does not appear to be a major factor in the decline of leopard frogs and boreal toads
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
Hamer, Gabriel L; Anderson, Tavis K; Donovan, Danielle J; Brawn, Jeffrey D; Krebs, Bethany L; Gardner, Allison M; Ruiz, Marilyn O; Brown, William M; Kitron, Uriel D; Newman, Christina M; Goldberg, Tony L; Walker, Edward D
2014-03-01
Dispersal is a critical life history behavior for mosquitoes and is important for the spread of mosquito-borne disease. We implemented the first stable isotope mark-capture study to measure mosquito dispersal, focusing on Culex pipiens in southwest suburban Chicago, Illinois, a hotspot of West Nile virus (WNV) transmission. We enriched nine catch basins in 2010 and 2011 with 15N-potassium nitrate and detected dispersal of enriched adult females emerging from these catch basins using CDC light and gravid traps to distances as far as 3 km. We detected 12 isotopically enriched pools of mosquitoes out of 2,442 tested during the two years and calculated a mean dispersal distance of 1.15 km and maximum flight range of 2.48 km. According to a logistic distribution function, 90% of the female Culex mosquitoes stayed within 3 km of their larval habitat, which corresponds with the distance-limited genetic variation of WNV observed in this study region. This study provides new insights on the dispersal of the most important vector of WNV in the eastern United States and demonstrates the utility of stable isotope enrichment for studying the biology of mosquitoes in other disease systems.
Papaefthimiou, Chrisovalantis; Zafeiridou, Georgia; Topoglidi, Aglaia; Chaleplis, George; Zografou, Stella; Theophilidis, George
2003-07-01
Three triazine herbicides, atrazine, simazine and metribuzine, and some of their major metabolites (cyanuric acid and 6-azauracil) were investigated for their action on synaptic terminals using three different isolated tissue preparations from the atria of the frog, Rana ridibunda, the heart of the honeybee, Apis mellifera macedonica, and the ventral nerve cord of the beetle, Tenebrio molitor. The results indicate that triazines facilitate the release of neurotransmitters from nerve terminals, as already reported for the mammalian central nervous system. The no observed effect concentration, the maximum concentration of the herbicide diluted in the saline that has no effect on the physiological properties of the isolated tissue, was estimated for each individual preparation. According to their relative potency, the three triazines tested can be ranked as follows: atrazine (cyanuric acid), simazine>metribuzine (6-azauracil). The action of these compounds on the cholinergic (amphibians, insects), adrenergic (amphibian) and octopaminergic (insects) synaptic terminals is discussed.
Kyek, Martin; Lindner, Robert
2017-01-01
This study focuses on the population trends of two widespread European anuran species: the common toad (Bufo bufo) and the common frog (Rana temporaria). The basis of this study is data gathered over two decades of amphibian fencing alongside roads in the Austrian state of Salzburg. Different statistical approaches were used to analyse the data. Overall average increase or decrease of each species was estimated by calculating a simple average locality index. In addition the statistical software TRIM was used to verify these trends as well as to categorize the data based on the geographic location of each migration site. The results show differing overall trends for the two species: the common toad being stable and the common frog showing a substantial decline over the last two decades. Further analyses based on geographic categorization reveal the strongest decrease in the alpine range of the species. Drainage and agricultural intensification are still ongoing problems within alpine areas, not only in Salzburg. Particularly in respect to micro-climate and the availability of spawning places these changes appear to have a greater impact on the habitats of the common frog than the common toad. Therefore we consider habitat destruction to be the main potential reason behind this dramatic decline. We also conclude that the substantial loss of biomass of a widespread species such as the common frog must have a severe, and often overlooked, ecological impact. PMID:29121054
Duerr, Jeffrey M; Tucker, Kristina
2007-08-01
Cardiac mitochondria were isolated from Bufo marinus and Rana catesbeiana, two species of amphibian whose cardiovascular systems are adapted to either predominantly aerobic or glycolytic modes of locomotion. Mitochondrial oxidative capacity was compared using VO2 max and respiratory control ratios in the presence of a variety of substrates including pyruvate, lactate, oxaloacetate, beta-hydroxybutyrate, and octanoyl-carnitine. B. marinus cardiac mitochondria exhibited VO2 max values twice that of R. catesbeiana cardiac mitochondria when oxidizing carbohydrate substrates. Pyruvate transport was measured via a radiolabeled-tracer assay in isolated B. marinus and R. catesbeiana cardiac mitochondria. Time-course experiments described both alpha-cyano-4-hydroxycinnamate-sensitive (MCT-like) and phenylsuccinate-sensitive pyruvate uptake mechanisms in both species. Pyruvate uptake by the MCT-like transporter was enhanced in the presence of a pH gradient, whereas the phenylsuccinate-sensitive transporter was inhibited. Notably, anuran cardiac mitochondria exhibited activities of lactate dehydrogenase and pyruvate carboxylase. The presence of both transporters on the inner mitochondrial membrane affords the net uptake of monocarboxylates including pyruvate, beta-hydroxybutyrate, and lactate; the latter potentially indicating the presence of a lactate/pyruvate shuttle allowing oxidation of extramitochondrial NADH. Intramitochondrial lactate dehydrogenase and pyruvate carboxylase enables lactate to be oxidized to pyruvate or converted to anaplerotic oxaloacetate. Kinetics of the MCT-like transporter differed significantly between the two species, suggesting differences in aerobic scope may be in part attributable to differences in mitochondrial carbohydrate utilization. (c) 2007 Wiley-Liss, Inc.
Wagner, Norman; Veith, Michael; Lötters, Stefan; Viertel, Bruno
2017-01-01
Environmental contamination is suggested to contribute to amphibian population declines. However, the effects of a contaminant on a particular amphibian species can differ among populations. The authors investigated the toxic effects of 2 herbicide formulations on different populations and on representative developmental stages of the European common frog (Rana temporaria). Larvae from forest populations were more sensitive to a commonly used glyphosate-based herbicide compared with individuals from agrarian land. Median lethal concentrations correlated with measured glyphosate levels in the breeding ponds, which may be a sign of evolved tolerances. The reverse result was observed for a less commonly used cycloxydim-based herbicide. Effects of the glyphosate-based herbicide were stronger for earlier larval stages compared with later larval stages. Hence, applications in early spring (when early larvae are present in breeding ponds) pose greater risk concerning acute toxic effects on R. temporaria. With regard to late larval stages, short exposure (96 h) of prometamorphic larvae prolonged time to metamorphosis, but only at the highest test concentration that did not significantly induce mortality. This could be due to impairment of the thyroid axis. Notably, nearly all test concentrations of the 2 herbicides provoked growth retardation. Further research on how evolved or induced tolerances are acquired, actual contamination levels of amphibian habitats, and potential endocrine effects of glyphosate-based herbicides is necessary. Environ Toxicol Chem 2017;36:190-200. © 2016 SETAC. © 2016 SETAC.
López-Torres, M; Pérez-Campo, R; Barja de Quiroga, G
1991-01-01
A study of the physiological role of oxygen free radicals in relation to the ageing process was performed using the liver of Rana perezi, an animal with a moderate rate of oxygen consumption and a life span substantially longer than that of laboratory rodents. Among the five different antioxidant enzymes only superoxide dismutase (SOD) showed an age-dependent decrease. Cytochrome oxidase (COX), glutathione status, in vivo and in vitro liver peroxidation, and metabolic rate did not vary as a function of age. Long-term (2.5 months) treatment with aminotriazole and diethyldithiocarbamate depleted catalase (CAT) activity and did not change both glutathione peroxidases (GPx), COX, reduced (GSH) and oxidized (GSSG) glutathione, or metabolic rate. This treatment resulted in great compensatory increases in SOD (to 250-460% of controls) and glutathione reductase (GR) (to 200%) which are possibly responsible for the lack of increase of in vivo and in vitro liver peroxidation and for the absence of changes in survival rate. The comparison of these results with previous data from other species suggests the possibility that decreases in antioxidant capacity in old age are restricted to animal species with high metabolic rates. Nevertheless, ageing can still be due to the continuous presence of small concentrations of O2 radicals in the tissues throughout life in animals with either high or low metabolic rates, because radical scavenging can not be 100% effective. Compensatory homeostasis among antioxidants seems to be a general phenomenon in different species.
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.
Avian Plasmodium in Eastern Austrian mosquitoes.
Schoener, Ellen; Uebleis, Sarah Susanne; Butter, Julia; Nawratil, Michaela; Cuk, Claudia; Flechl, Eva; Kothmayer, Michael; Obwaller, Adelheid G; Zechmeister, Thomas; Rubel, Franz; Lebl, Karin; Zittra, Carina; Fuehrer, Hans-Peter
2017-09-29
Insect vectors, namely mosquitoes (Diptera: Culicidae), are compulsory for malaria parasites (Plasmodium spp.) to complete their life cycle. Despite this, little is known about vector competence of different mosquito species for the transmission of avian malaria parasites. In this study, nested PCR was used to determine Plasmodium spp. occurrence in pools of whole individuals, as well as the diversity of mitochondrial cytochrome b gene sequences in wild-caught mosquitoes sampled across Eastern Austria in 2013-2015. A total of 45,749 mosquitoes in 2628 pools were collected, of which 169 pools (6.43%) comprising 9 mosquito species were positive for avian Plasmodium, with the majority of positives in mosquitoes of Culex pipiens s.l./Culex torrentium. Six different avian Plasmodium lineages were found, the most common were Plasmodium vaughani SYAT05, Plasmodium sp. Linn1 and Plasmodium relictum SGS1. In 2014, mosquitoes of the Culex pipiens complex were genetically identified and Culex pipiens f. pipiens presented with the highest number of avian Plasmodium positives (n = 37; 16.74%). Despite this, the minimum infection rate (MIR) was highest in Culex torrentium (5.36%) and Culex pipiens f. pipiens/f. molestus hybrids (5.26%). During 2014 and 2015, seasonal and annual changes in Plasmodium lineage distribution were also observed. In both years P. vaughani SYAT05 dominated at the beginning of the sampling period to be replaced later in the year by P. relictum SGS1 (2014) and Plasmodium sp. Linn1 (2015). This is the first large-scale study of avian Plasmodium parasites in Austrian mosquitoes. These results are of special interest, because molecular identification of the taxa of the Cx. pipiens complex and Cx. torrentium enabled the determination of Plasmodium prevalence in the different mosquito taxa and hybrids of this complex. Since pools of whole insects were used, it is not possible to assert any vector competence in any of the examined mosquitoes, but the results
Rocha, P L; Branco, L G
1998-03-01
We assessed seasonal variations in the effects of temperature on hypoxia-induced alterations in the bullfrog Rana catesbeiana by measuring the heart rate, arterial blood pressure, breathing frequency, metabolic rate, blood gas levels, acid-base status and plasma glucose concentration. Regardless of the season, decreased body temperature was accompanied by a reduction in heart and breathing frequencies. Lower temperatures caused a significant decrease in arterial blood pressure during all four seasons. Hypoxia-induced changes in breathing frequency were proportional to body temperature and were more pronounced during winter, less so during spring and autumn and even smaller during summer. Season had no effect on the relationship between hypoxia and heart rate. At any temperature tested, the rate of oxygen consumption had a tendency to be highest during summer and lowest during winter, but the difference was significant only at 35 degrees C. The PaO2 and pH values showed no significant change during the year, but PaCO2 was almost twice as high during winter than in summer and spring, indicating increased plasma bicarbonate levels. Lower temperatures were accompanied by decreased plasma glucose levels, and this effect was greater during summer and smaller during autumn. Hypoxia-induced hyperglycaemia was influenced by temperature and season. During autumn and winter, plasma glucose level remained elevated regardless of temperature, probably to avoid dehydration and/or freezing. In winter, the bullfrog may be exposed not only to low temperatures but also to hypoxia. These animals show temperature-dependent responses that may be beneficial since at low body temperatures the set-points of most physiological responses to hypoxia are reduced, regardless of the season.
Sulesco, Tatiana M; Toderas, Lidia G; Uspenskaia, Inga G; Toderas, I K
2015-11-01
A countrywide field survey of immature mosquitoes was conducted in Moldova with the aim to evaluate the Culicidae species composition in different larval habitats and their distribution in the country. In total, 259 potential larval habitats were sampled in the 53 localities, resulting in 9,456 specimens. Twenty species belonging to the genera Anopheles, Aedes, Culex, Culiseta, and Uranotaenia were collected. Mean species richness in aquatic habitats ranged from 1.00 to 4.00, and, for example, was higher in swamps, flood plains, ditches, and large ground pools and lower in rivers, streams, tree-holes, and containers. Six mosquito species were identified only in a single type of aquatic habitat. Anopheles maculipennis s.l., Culex pipiens pipiens L., and Culex modestus Ficalbi were the most abundant and distributed species representing over 80% of the identified specimens. Three, four, and five associated species were recorded from 23.5% of mosquito-positive aquatic habitats. Our findings demonstrate the co-occurrence of Cx. p. pipiens and Culex torrentium Martini in natural and rural environments. It is concluded that the study area has undergone a dramatic ecological change since the previous studies in the 1950s, causing the near extinction of Culex theileri Theobald from Moldova. An. maculipennis s.l. larval abundance, reduced by the DDT control of the adults in the 1950s, had returned to those of the 1940s. Restoration of An. maculipennis s.l. abundance in combination with imported malaria cases constitute a risk of the reintroduction of malaria transmission in Moldova. © The Authors 2015. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.
Navas, Carlos A; James, Rob S
2007-02-01
Mating success of individual male frogs within explosive breeding species can depend on their ability to compete for a mate and to hold onto that mate during amplexus. Such importance of amplexus has resulted in the evolution of sexual dimorphism in the morphology and contractile characteristics of the anuran forelimb muscles used during amplexus. The aims of our study were to use an explosive breeding frog (Rana temporaria) during the breeding season to compare extensor carpi radialis (ECR) muscle length, mass, isometric activation times, relaxation times, absolute force, relative force (stress) and fatigue between male and female frogs. We found that ECR muscle mass and length were greater (tenfold and 1.4-fold, respectively), absolute tetanic muscle force and relative tetanic force (stress) were greater (16-fold and 2.2-fold, respectively) and relaxation times were slower in males than in females. Male ECR muscles incompletely relaxed during fatigue tests and showed less fatigue than female muscles. These sex differences are likely to be beneficial to the male frogs in allowing them to produce relatively high absolute muscle forces for prolonged periods of time to hold onto their mate during amplexus.
Tavazoie, S F; Tavazoie, M F; McIntosh, J M; Olivera, B M; Yoshikami, D
1997-03-01
1. The effects of two new acetylcholine receptor antagonists, alpha-conotoxin MII and alpha-conotoxin ImI, on nicotinic synaptic transmission in the 10th paravertebral sympathetic ganglion of the leopard frog (Rana pipiens) were examined. The preganglionic nerve was electrically stimulated (at low frequency, < or = 1 min-1, to avoid use-dependent changes) while compound action potentials of B and C neurones were monitored from the postganglionic nerve. 2. alpha-Conotoxins MII and ImI, at low micromolar concentrations, reversibly blocked both B and C waves, alpha-Conotoxin MII blocked the C wave more effectively than the B wave, whereas the potency of alpha-conotoxin ImI was opposite that of MII. The observation that nicotinic antagonists can differentially block synaptic transmission of B versus C neurones with opposite selectivities strongly suggests that these neurones possess distinct nicotinic receptors. 3. In addition to fast and slow B waves described by others. C waves with two temporally distinguishable components were present in our recordings. Each alpha-conotoxin affected fast and slow B waves similarly. Likewise, toxins did not discriminate between the two components of C waves. This suggests that all neurones within each major class (B or C) may have the same nicotinic receptors. 4. Synthetic forms of alpha-conotoxins MII and ImI were used in the present study. Their ease of synthesis and their specificities should make these toxins useful probes to investigate the various subtypes of neuronal nicotinic acetylcholine receptors.
DOE Office of Scientific and Technical Information (OSTI.GOV)
Pentecost, E. D.; Stupka, R. C.
Wildlife studies of a reclaimed coal refuse pile were conducted during the summer months of 1977 and 1978. During the period of May--September 1977 and May--July 1978 live-trap censuses were made. Small mammal species observed in 1977 included the white-footed mouse (Peromyscus leucopus), house mouse (Mus musculas) and meadow vole (Microtus pennsylvanicus). In 1978 the prairie vole (Microtus ochrogaster) was also a common resident of the site. Population density estimates for Mus musculus and M. ochrogaster were 20 (+-19.4) and 37.5 (+-20) respectively in June 1978. Evidence of habitat segregation was observed between M. pennsylvanicus and M. ochrogaster with themore » former species occupying densely vegetated areas of tall fescue (Festuca arundinaces), birdsfoot trefoil (Phalaris arundinacea) and cereal rye (Secale cereale) while M. ochrogaster was more abundant in open areas with similar vegetation. Both the mammalian and avian species observed on site were those typical of early old-field communities in southern Illinois. Four passerine species (kill-deer, song sparrow, red-winged blackbird, common grackle) nested on site during the second year following seeding. No onsite nesting activity occurred during 1977. Common amphibians observed around the pond included the southern leopard frog (Rana pipiens sphenocephala), Blanchards' cricket frog (Acris crepitans blanchardi) and american toad (Bufo a. americanus). Utilization of the pond by these species occurred the first summer after reclamation was completed.« less
In Vivo Confocal Intrinsic Optical Signal Identification of Localized Retinal Dysfunction
Zhang, Qiu-Xiang; Lu, Rong-Wen; Curcio, Christine A.; Yao, Xin-Cheng
2012-01-01
Purpose. The purposes of this study were to investigate the physiological mechanism of stimulus-evoked fast intrinsic optical signals (IOSs) recorded in dynamic confocal imaging of the retina, and to demonstrate the feasibility of in vivo confocal IOS mapping of localized retinal dysfunctions. Methods. A rapid line-scan confocal ophthalmoscope was constructed to achieve in vivo confocal IOS imaging of frog (Rana pipiens) retinas at cellular resolution. In order to investigate the physiological mechanism of confocal IOS, comparative IOS and electroretinography (ERG) measurements were made using normal frog eyes activated by variable-intensity stimuli. A dynamic spatiotemporal filtering algorithm was developed to reject the contamination of hemodynamic changes on fast IOS recording. Laser-injured frog eyes were employed to test the potential of confocal IOS mapping of localized retinal dysfunctions. Results. Comparative IOS and ERG experiments revealed a close correlation between the confocal IOS and retinal ERG, particularly the ERG a-wave, which has been widely used to evaluate photoreceptor function. IOS imaging of laser-injured frog eyes indicated that the confocal IOS could unambiguously detect localized (30 μm) functional lesions in the retina before a morphological abnormality is detectable. Conclusions. The confocal IOS predominantly results from retinal photoreceptors, and can be used to map localized photoreceptor lesion in laser-injured frog eyes. We anticipate that confocal IOS imaging can provide applications in early detection of age-related macular degeneration, retinitis pigmentosa, and other retinal diseases that can cause pathological changes in the photoreceptors. PMID:23150616
Palma, F; Roncagliolo, P; Bacigalupo, J; Palacios, A G
2001-01-01
We investigated the photocurrents from isolated rods of the South American anuran, Caudiverbera caudiverbera. Rod outer segments were on average 66.4 +/- 11.2 microm (mean +/- S.D., n = 104) in length and 6.6 +/- 0.9 microm (mean +/- S.D.) in diameter: 40 +/- 22 photoisomerizations (mean +/- S.D., range 10-99, n = 16) were required for eliciting a half-saturating photocurrent response. The time-to-peak was 911 +/- 217 ms (mean +/- S.D., n = 14, 20 degrees C) in the linear range of the response and the integration time of the current response was 1744 +/- 451 ms (mean +/- S.D., n = 14). The time-to-peak appears to be slower and the integration time shorter in Caudiverbera than in Ambystoma tigrinum, Rana pipiens or Xenopus laevis rods under similar experimental conditions. The a-band of rod spectral sensitivity has a lambda(max) at 520 +/- 2.1 nm (mean +/- S.D., range 516-525 nm, n = 24) and the bandwidth fits a porphyropsin visual pigment. The single-event response amplitude ranges from 0.31-0.51 pA, depending on the calculation method. The intrinsic dark current (variance at dark minus variance under bright light) was 0.045 +/- 0.040 pA2 (mean +/- S.D., n = 24). Our results support the presence of a dark-noise component below 1 Hz, with kinetics similar to the single-photon evoked response and a rate of 0.006 events s(-1) (n = 9).
Analysis of isolates within species of anuran trypanosomes using random amplified polymorphic DNA.
Lun, Z R; Desser, S S
1996-01-01
A total of 20 decamer primers were used to generate random applied polymorphic DNA (RAPD) markers from 5 isolates of Trypanosoma fallisi, 3 isolates of T. ranarum, 2 isolates of T. rotatorium, and 2 isolates of T. rotatorium-like trypanosomes in addition to 2 species from the American Type Culture Collection, T. chattoni (ATCC 50294) and Trypanosoma sp. (ATCC 50295). A slight polymorphism was observed among the four isolates of T. fallisi obtained form American toads, Bufo americanus, collected in Algonquin Park, Ontario, Canada, and an isolate obtained from the same species of host collected in Marquette, Michigan, United States, and produced similarity coefficients ranging from 80.7% to 96.9%. Pronounced polymorphism was recorded among the three isolates of T. ranarum from bullfrogs, Rana catesbeiana, collected in Ontario, Canada, and in Maryland, United States, and from a Northern leopard frog, R. pipiens, collected in Minnesota (USA). The similarity coefficients ranged from 54.7% to 59.5%, suggesting that alleles of these isolates were conserved over a wide geographic range. The high degree of polymorphism observed in two isolates of T. rotatorium from a bullfrog collected in Ontario and two isolates of a T. rotatorium-like parasite from the green frog R. clamitans, collected in Louisiana (USA) suggests that they are different species. These results reflect the high similarity among isolates from the same geographic location and the pronounced polymorphism apparent among isolates from distant geographic locations.
USDA-ARS?s Scientific Manuscript database
We examined the chemical composition of garlic and asafoetida essential oils and their individual and combined toxicity against larvae of two West Nile virus vectors, Culex pipiens pipiens and Cx. restuans. The effect of the two essential oils on egg hatch was also examined. Ten and twelve compounds...
NELMS, BRITTANY M.; MACEDO, PAULA A.; KOTHERA, LINDA; SAVAGE, HARRY M.; REISEN, WILLIAM K.
2014-01-01
At temperate latitudes, Culex (Diptera: Culicidae) mosquitoes typically overwinter as adult females in reproductive arrest and also may serve as reservoir hosts for arboviruses when cold temperatures arrest viral replication. To evaluate their role in the persistence of West Nile virus (WNV) in the Sacramento Valley of California, the induction and termination of diapause were investigated for members of the Culex pipiens (L.) complex, Culex tarsalis Coquillett, and Culex stigmatosoma Dyar under field, seminatural, and experimental conditions. All Culex spp. remained vagile throughout winter, enabling the collection of 3,174 females and 1,706 males from diverse habitats during the winters of 2010–2012. Overwintering strategies included both quiescence and diapause. In addition, Cx. pipiens form molestus Forskäl females remained reproductively active in both underground and aboveground habitats. Some blood-fed, gravid, and parous Cx. tarsalis and Cx. pipiens complex females were collected throughout the winter period. Under both field and experimental conditions, Cx. tarsalis and Cx. stigmatosoma females exposed to autumnal conditions arrested primary follicular maturation at previtellogenic stage I, with primary to secondary follicular ratios <1.5 (indicative of a hormonally induced diapause). In contrast, most Cx. pipiens complex females did not enter reproductive diapause and ovarian follicles matured to ≥stage I–II (host-seeking arrest) or were found in various stages of degeneration. Diapause was initiated in the majority of Cx. tarsalis and Cx. stigmatosoma females by mid-late October and was terminated after the winter solstice, but host-seeking seemed limited by temperature. An accrual of 97.52 ± 30.7 and 162.85 ± 79.3 degree-days after the winter solstice was estimated to be necessary for diapause termination in Cx. tarsalis under field and seminatural conditions, respectively. An increase in the proportion of blood-fed Culex females in resting
Identifying homomorphic sex chromosomes from wild-caught adults with limited genomic resources.
Brelsford, Alan; Lavanchy, Guillaume; Sermier, Roberto; Rausch, Anna; Perrin, Nicolas
2017-07-01
We demonstrate a genotyping-by-sequencing approach to identify homomorphic sex chromosomes and their homolog in a distantly related reference genome, based on noninvasive sampling of wild-caught individuals, in the moor frog Rana arvalis. Double-digest RADseq libraries were generated using buccal swabs from 30 males and 21 females from the same population. Search for sex-limited markers from the unfiltered data set (411 446 RAD tags) was more successful than searches from a filtered data set (33 073 RAD tags) for markers showing sex differences in heterozygosity or in allele frequencies. Altogether, we obtained 292 putatively sex-linked RAD loci, 98% of which point to male heterogamety. We could map 15 of them to the Xenopus tropicalis genome, all but one on chromosome pair 1, which seems regularly co-opted for sex determination among amphibians. The most efficient mapping strategy was a three-step hierarchical approach, where R. arvalis reads were first mapped to a low-coverage genome of Rana temporaria (17 My divergence), then the R. temporaria scaffolds to the Nanorana parkeri genome (90 My divergence), and finally the N. parkeri scaffolds to the X. tropicalis genome (210 My). We validated our conclusions with PCR primers amplifying part of Dmrt1, a candidate sex determination gene mapping to chromosome 1: a sex-diagnostic allele was present in all 30 males but in none of the 21 females. Our approach is likely to be productive in many situations where biological samples and/or genomic resources are limited. © 2016 John Wiley & Sons Ltd.
Mweya, Clement N; Kimera, Sharadhuli I; Mellau, Lesakit S B; Mboera, Leonard E G
2015-01-01
Rift Valley fever (RVF) is a mosquito-borne viral zoonosis that primarily affects ruminants but also has the capacity to infect humans. To determine the abundance and distribution of mosquito vectors in relation to their potential role in the virus transmission and maintenance in disease epidemic areas of Ngorongoro district in northern Tanzania. A cross-sectional entomological investigation was carried out before the suspected RVF outbreak in October 2012. Mosquitoes were sampled both outdoors and indoors using the Centre for Disease Control (CDC) light traps and Mosquito Magnets baited with attractants. Outdoor traps were placed in proximity with breeding sites and under canopy in banana plantations close to the sleeping places of animals. A total of 1,823 mosquitoes were collected, of which 87% (N=1,588) were Culex pipiens complex, 12% (N=226) Aedes aegypti, and 0.5% (N=9) Anopheles species. About two-thirds (67%; N=1,095) of C. pipiens complex and nearly 100% (N=225) of A. aegypti were trapped outdoors using Mosquito Magnets. All Anopheles species were trapped indoors using CDC light traps. There were variations in abundance of C. pipiens complex and A. aegypti among different ecological and vegetation habitats. Over three quarters (78%) of C. pipiens complex and most (85%) of the A. aegypti were trapped in banana and maize farms. Both C. pipiens complex and A. aegypti were more abundant in proximity with cattle and in semi-arid thorn bushes and lower Afro-montane. The highest number of mosquitoes was recorded in villages that were most affected during the RVF epidemic of 2007. Of the tested 150 pools of C. pipiens complex and 45 pools of A. aegypti, none was infected with RVF virus. These results provide insights into unique habitat characterisation relating to mosquito abundances and distribution in RVF epidemic-prone areas of Ngorongoro district in northern Tanzania.
Ruiz-Torres, Nallely; García-Prieto, Luis; Osorio-Sarabia, David; Violante-González, Juan
2013-06-01
Oswaldocruzia lamotheargumedoi n. sp., inhabiting the intestine of the cane toad, Rhinella marina (L.), in Laguna de Coyuca, Guerrero, México, is described here. The new species differs from 10 congeners infecting bufonid hosts because it has a type I bursa. In contrast, 7 of these species have type II bursa and 3 more a type III bursa. The species most similar to the species described herein is Oswaldocruzia pipiens Walton, 1929 . These 2 species share traits such as body size, bursa type, presence of cervical alae, and dorsal ray morphology. Nevertheless, both species can be distinguished based on the number of synlophe ridges at mid-body (54-56 for O. lamotheargumedoi vs. 45-48 for O. pipiens) and by the presence of a chitinous support in the long, and well developed, cervical alae of O. pipiens. In the new species, these structures are short, poorly developed, and lack chitinous support. Previous records of species of Oswaldocruzia in México include Oswaldocruzia subauricularis (Rudolphi, 1819) Travassos, 1917 in the Neotropical Realm and O. pipiens in the Nearctic.
Aliota, Matthew T.; Jones, Susan A.; Dupuis, Alan P.; Ciota, Alexander T.; Hubalek, Zdenek; Kramer, Laura D.
2012-01-01
Rabensburg virus (RABV), a Flavivirus with ∼76% nucleotide and 90% amino acid identity with representative members of lineage one and two West Nile virus (WNV), previously was isolated from Culex pipiens and Aedes rossicus mosquitoes in the Czech Republic, and phylogenetic and serologic analyses demonstrated that it was likely a new lineage of WNV. However, no direct link between RABV and human disease has been definitively established and the extent to which RABV utilizes the typical WNV transmission cycle is unknown. Herein, we evaluated vector competence and capacity for vertical transmission (VT) in Cx. pipiens; in vitro growth on avian, mammalian, and mosquito cells; and infectivity and viremia production in birds. RABV infection and replication only were detected on mosquito cells. Experimentally inoculated birds did not become infected. Cx. pipiens had poor peroral vector competence and a higher VT rate as compared to US-WNV in Cx. pipiens. As a result, we postulate that RABV is an intermediate between the mosquito-specific and horizontally transmitted flaviviruses. PMID:22724010
Experimental transmission of St. Louis encephalitis virus by Ochlerotatus j. japonicus.
Sardelis, Michael R; Turell, Michael J; Andre, Richard G
2003-06-01
Ochlerotatus japonicus japonicus a newly discovered nonindigenous mosquito species in North America, and a colonized strain of Culex pipiens were compared for their vector competence for St. Louis encephalitis virus (SLE). Infection rates in Oc. j. japonicus were 0-33% after feeding on chickens with viremias between 10(4.1) and 10(4.7) plaque-forming units (PFU)/ml of blood. In comparison, infection rates were 12-94% for Cx. pipiens that fed on the same chickens. When fed on chickens with viremias between 10(5.3) and 10(5.6) PFU/ml of blood, infection rates for Oc. j. japonicus and Cx. pipiens were similar, 96% and 100%, respectively. After 12-14 days of extrinsic incubation at 26 degrees C, all 34 infected Oc. j. japonicus had a disseminated infection. In contrast, only 23 (43%) of 54 infected Cx. pipiens had a disseminated infection after feeding on the same chickens. If they developed a disseminated infection, both species efficiently transmitted (> or = 87%) SLE. Estimated transmission rates at viral doses sufficient to infect both of the tested species were 29-84% for Oc. j. japonicus and 30-50% for Cx. pipiens. Because of its continued geographic expansion, field and laboratory evidence incriminating it as a vector of the closely related West Nile virus, and its ability to transmit SLE in the laboratory, Oc. j. japonicus should be considered as a potential enzootic or epizootic vector of SLE.
Kühn, E R; Delmotte, N M; Darras, V M
1983-06-01
The presence and circadian rhythmicity of thyroid hormones was studied in plasma and the thyroid gland of male Rana ridibunda before and during hibernation. Hibernating January frogs do have a lower T3 and T4 content of their thyroid gland whereas plasma levels of T3 are maintained and of T4 increased compared to fed September or October frogs. It seems likely that the increased photoperiod in January will be responsible for this increased T4 secretion, since controlled laboratory experiments performed in December did not reveal any influence of low temperature on circulating T3 or T4 levels. Also feeding does not influence circulating levels and thyroid content of thyroid hormones in frogs kept at room temperature during the month of January. A circadian rhythmicity of T3 and T4 in the thyroid gland is present in fed October frogs and in non fed December frogs acclimated at 5 degrees C for 12 days with an acrophase for T3 at approximately 1500 h and for T4 at around 1900 h, whereas in plasma only T3 does have circadian variations (acrophase about midnight) but not T4. When December frogs are acclimated to room temperature for 12 days, frogs are active again, but do not eat and have a lower body weight than frogs hibernating at 5 degrees C. Their T3 content of the thyroid gland has disappeared, but T4 thyroid content and plasma levels of T3 and T4 are maintained. As in hibernating frogs, no circadian variations in T4 plasma concentrations are present whereas the circadian thyroid T4 rhythm disappears. At the same time a dampening in rhythmicity for plasma T3 as judged by the significantly lower amplitude occurs. It is concluded that the persistence of circulating levels of thyroid hormones and of a circadian cyclicity for T3 in plasma in non feeding hibernating frogs may reflect the special metabolic state e.g. availability of food reserves in these animals.
Mashreghi, Mohammad; Rezazade Bazaz, Mahere; Mahdavi Shahri, Nasser; Asoodeh, Ahmad; Mashreghi, Mansour; Behnam Rassouli, Morteza; Golmohammadzadeh, Shiva
2013-02-13
Study of the interrelationships between human and the animals in their environment has always been a subject of interest and caused discoveries of the animal applications in medicine. From the latest century, these remedies called back in traditional medicine of Vietnam and South America and frog skin was used as a biological dressing and had good effects in healing wounds. Also, frog skin secretions have wound healing properties and reduce inflammation. In this study we applied these secretions in the form of ointment to investigate their healing activities. Skin secretions were extracted from Rana ridibunda to evaluate their effects on wound healing in mice. Secretion used as raw extract (RE) and ultrafiltrated extract, using a membrane with cutoff 10kDa as under 10kDa (U10E), was administrated as ointment every 48h on wound site. Control group was left without any treatment and also there was other group treated with ointment (O group) alone. On 2, 4 and 6 days post injury, animals were euthanized and images were taken for wound closure evaluation. Then wound locations were removed for histological assays. Also wound microbial load was examined. Observational parameters including wound closure and wound microbiology in experimental groups compared with the control and O groups have been studied. The results showed U10E group has better effects than RE, O and control groups. Histological parameters, including numbers of inflammatory and fibroblast cells and amount of collagen fibers, neovascularization, as well, represented greater degree of wound healing in U10E group compared with RE, O, and control groups. Our results showed that frog skin secretions were significantly effective in promoting wound healing process. The U10E extract from the frog R. ridibunda possesses a potent accelerating wound healing effect that promises good potential for clinical application in wound care. Further studies will be required to characterize special molecules encompassing
Hoyeck, Myriam P.; Hadj-Moussa, Hanane
2017-01-01
The wood frog (Rana sylvatica) can endure freezing of up to 65% of total body water during winter. When frozen, wood frogs enter a dormant state characterized by a cessation of vital functions (i.e., no heartbeat, blood circulation, breathing, brain activity, or movement). Wood frogs utilize various behavioural and biochemical adaptations to survive extreme freezing and component anoxia and dehydration stresses, including a global suppression of metabolic functions and gene expression. The stress-responsive myocyte enhancer factor-2 (MEF2) transcription factor family regulates the selective expression of genes involved in glucose transport, protein quality control, and phosphagen homeostasis. This study examined the role of MEF2A and MEF2C proteins as well as select downstream targets (glucose transporter-4, calreticulin, and muscle and brain creatine kinase isozymes) in 40% dehydration and 24 h anoxia exposure at the transcriptional, translational, and post-translational levels using qRT-PCR, immunoblotting, and subcellular localization. Mef2a/c transcript levels remained constant during dehydration and anoxia. Total, cytoplasmic, and nuclear MEF2A/C and phospho-MEF2A/C protein levels remained constant during dehydration, whereas a decrease in total MEF2C levels was observed during rehydration. Total and phospho-MEF2A levels remained constant during anoxia, whereas total MEF2C levels decreased during 24 h anoxia and P-MEF2C levels increased during 4 h anoxia. In contrast, cytoplasmic MEF2A levels and nuclear phospho-MEF2A/C levels were upregulated during anoxia. MEF2 downstream targets remained constant during dehydration and anoxia, with the exception of glut4 which was upregulated during anoxia. These results suggest that the upregulated MEF2 response reported in wood frogs during freezing may in part stem from their cellular responses to surviving prolonged anoxia, rather than dehydration, leading to an increase in GLUT4 expression which may have an important
Federal Register 2010, 2011, 2012, 2013, 2014
2013-10-01
... prevent the extinction of two species of mountain yellow-legged frogs (Rana muscosa and Rana sierrae; MYLF... enable the NPS to fulfill its mission and policy directives to conserve native animals, plants and...
Li, Long; Wang, Wenyue; Lv, Quanxia; Ben, Yujie; Li, Xinghong
2014-03-01
Dechlorane Plus (DP), a flame retardant used as an alternative to decabromodiphenylether, has been frequently detected in organisms, indicating its bioaccumulation and biomagnification potential in aquatic and terrestrial species. However, little data is available on the bioaccumulation of DP in amphibians. Dechlorane Plus and its analogs (DPs) were detected in the liver, muscle and brain tissues of wild frogs (Rana limnocharis), which were collected from an e-waste recycling site, Southeast China. DP, Mirex, Dec 602 and a dechlorinated compound of DP (anti-Cl11-DP) varied in the range of 2.01-291, 0.650-179, 0.260-12.4, and not detected (nd)-8.67 ng/g lipid weight, respectively. No difference of tissue distribution was found for syn-DP, Mirex and Dec 602 between the liver and muscle tissue (liver/muscle concentration ratio close to 1, p > 0.05). However, higher retention was observed for anti-DP and anti-Cl11-DP in the frog muscle relative to the liver tissue (liver/muscle concentration ratio < 1, p < 0.05). Additionally, the blood-brain barrier was found to work efficiently to suppress these compounds entering brain tissues in this species (liver/brain concentration ratio > 1, p < 0.05), and the molecular weight was a key factor impacting the extent of the blood-brain barrier. Compared to levels in the muscle and brain tissue, a preferential enrichment of syn-DP was observed in the liver tissue, suggesting the occurrence of stereo-selective bioaccumulation in the wild frog. Copyright © 2014 The Research Centre for Eco-Environmental Sciences, Chinese Academy of Sciences. Published by Elsevier B.V. All rights reserved.
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.
Nonanesthetic alcohols dissolve in synaptic membranes without perturbing their lipids.
Miller, K W; Firestone, L L; Alifimoff, J K; Streicher, P
1989-01-01
While many theories of general anesthesia postulate a lipid site of action, there has been no adequate explanation for the lack of anesthetic potency of the highly hydrophobic primary alkanols with more than 12 carbons (the cut-off). Some work suggests that these nonanesthetic alcohols do not dissolve in membranes. Other work contradicts this and suggests that an anesthetic site on a protein provides a better explanation. Here we show that both the anesthetic dodecanol and the nonanesthetic tetradecanol are taken up equally well into the tissues of animals and into isolated postsynaptic membranes. When a group of Rana pipiens tadpoles were treated with dodecanol, half were anesthetized by 4.7 microM (free aqueos concentration), and the corresponding concentration in the tissues was found to be 0.4 mmol per kg wet weight. Prolonged exposure (92 hr) to tetradecanol produced even higher tissue concentrations (0.7 mmol per kg wet weight), yet no anesthetic effects were observed. Furthermore, general anesthetics are thought to act on postsynaptic membranes but both alkanols partitioned into postsynaptic membranes from Torpedo electroplaques. The spin label, 12-doxyl stearate, was incorporated into these membranes. The lipid order parameter it reported was decreased by the anesthetic alcohols (octanol, decanol, and dodecanol), whereas the nonanesthetic alcohols either did not change it significantly (tetradecanol) or actually increased it (hexadecanol and octadecanol). Thus, although lipid solubility is unable to account for the pharmacology of the cut-off in potency of the long-chain alcohols, lipid perturbations provide an accurate description. PMID:2783782
Leopard frog PCB levels and evaluation of EROD as a biomarker in Green Bay ecosystem
DOE Office of Scientific and Technical Information (OSTI.GOV)
Huang, Y.W.; Karasov, W.H.; Patnode, K.P.
1995-12-31
The induction of mixed function oxidases has been shown to be a promising biomarker in many taxa of wildlife, though not yet tested for amphibians. The three hypotheses tested in this study were (1) activities of hepatic EROD of leopard frog (Rana pipiens) are induced following exposure to planar chlorinated PCBs, (2) tissue PCB residue levels of leopard frogs are positively correlated with their wetland sediment PCB levels, and (3) EROD activities are positively correlated with tissue PCB concentrations and sediment PCB. In the laboratory, EROD was increased 2--3 times seven days after i.p. injection with PCB 126 at dosesmore » {ge} 2.3 ppm (wet mass basis). Leopard frogs from seven sites along the Lower Fox River and Green Bay in 1994--1995 were assayed for hepatic EROD activities and total PCB levels in carcasses. Tissue PCB levels ranged from 3 to 152 ppb (including coplanar congeners) and were highest from sites with higher sediment PCB. EROD activity in frogs collected in August--September was not significantly correlated with frog body mass and was similar among sites with one exception. There was no significant correlation between EROD activity and tissue PCB concentration. This result was consistent with the fact that the frogs collected from the Green Bay ecosystem had relatively low PCB levels compared with what was required for induction in the laboratory. The authors conclude that EROD activity is not a sensitive biomarker of PCB exposure in leopard frogs in this ecosystem.« less
Fotakis, Emmanouil A; Chaskopoulou, Alexandra; Grigoraki, Linda; Tsiamantas, Alexandros; Kounadi, Stella; Georgiou, Loukas; Vontas, John
2017-10-01
Greece has been recently affected by several mosquito borne diseases with the West Nile Virus (WNV) outbreak in 2010 being one of the largest reported in Europe. Currently at the epicenter of an economic and refugee crisis and visited by over 16 million tourists a year the integrated management of diseases transmitted by mosquitoes is a public health and economic priority. Vector control programs rely mainly on insecticides, however data on insecticide resistance and the mosquito fauna is essential for successful applications. We determined the mosquito species composition and population dynamics in areas of increased vulnerability to vector borne disease transmission, as well as investigated the resistance status of major nuisance and disease vectors to insecticides. High mosquito densities were recorded in Thessaloniki and Evros, with Aedes caspius, a nuisance species, Culex pipiens, a known vector of WNV and Anopheles hyrcanus a potential vector of malaria being among the most prevalent species. Both vector species populations reached their peak in late summer. Aedes albopictus was recorded at high densities in Thessaloniki, but not in Evros. Notably, Cx. pipiens hybrids, which show an opportunistic biting behavior and are suspected to be involved in the transmission of the WNV, were recorded in considerable numbers in Thessaloniki and Attica. Culex pipiens and An. hyrcanus, but not Ae. caspius mosquitoes, showed moderate levels of resistance to deltamethrin. The presence of resistance in areas not exposed to vector control indicates that other factors could be selecting for resistance, i.e. pesticide applications for agriculture. Both L1014F and L101C kdr mutations were detected in Cx. pipiens populations. Anopheles hyrcanus resistance was not associated with mutations at the L1014 site. The Ace-1 mutations conferring insensitivity to organophosphates and carbamates were detected at low frequencies in all Cx. pipiens populations. Increased activity of P450s and
NASA Astrophysics Data System (ADS)
Lacan, I.; Matthews, K. R.
2005-12-01
Year to year variation in snowpack (20-200% average) and summer rain create large fluctuations in the volume of water in ponds and small lakes of the higher elevation (> 3000 m) Sierra Nevada. These water bodies are critical habitat for the imperiled mountain yellow-legged frog, Rana muscosa, which has decreased in abundance by 90% during the past century, due in part to the loss of suitable habitat and introduction of a fish predator (trout, Oncorhynchus spp.). Climate change is predicted to reduce the amount of snowpack, potentially impacting amphibian habitats throughout the Sierra Nevada by further reducing the lake and pond water levels and resulting in drying of small lakes during the summer. Mountain yellow-legged frogs are closely tied to water during all life stages, and are unique in having a three- to four-year tadpole phase. Thus, tadpole survival and future recruitment of adult frogs requires adequate water in lakes and ponds throughout the year, but larger lakes are populated with fish that prey on frogs and tadpoles. Thus, most successful frog breeding occurs in warm, shallow, fishless ponds that undergo wide fluctuations in volume. These water bodies would be most susceptible to the potential climate change effects of reduced snowpack, possibly resulting in lower tadpole survival. This study explores the link between the changes in water availability -- including complete pond drying -- and the abundance and recruitment of mountain yellow-legged frog in Dusy Basin, Kings Canyon National Park, California, USA. We propose using the low-snowpack years (1999, 2002, 2004) as comparative case studies to predict future effects of climate change on aquatic habitat availability and amphibian abundance and survival. To quantify the year to year variation and changes in water volume available to amphibians, we initiated GPS lake mapping in 2002 to quantify water volumes, water surface area, and shoreline length. We tracked these changes by repeated mapping of
Regulation of 5'-adenosine monophosphate deaminase in the freeze tolerant wood frog, Rana sylvatica.
Dieni, Christopher A; Storey, Kenneth B
2008-04-22
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. 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 approximately 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 degrees C) and by the presence/absence of the natural cryoprotectant (250 mM glucose) that accumulates during freezing
Mosquitoes of the genus Culex in the Suez Canal Governorates.
Morsy, T A; el Okbi, L M; Kamal, A M; Ahmed, M M; Boshara, E F
1990-06-01
Mosquitoes are among the most annoying and important vectors of human and animal diseases as malaria, filariasis, yellow fever, rift valley fever...etc. In this paper, it was aimed to study the present status of species of genus Culex in the Suez Canal Governorates after the reconstruction and developmental projects. Five species of Culex were identified: C. pipiens, C. univittatus, C. antennatus, C. poicilipes and C. pusillus. The latter species was represented by two specimens. C. pipiens was the commonest species both indoors and outdoors. C. antennatus and C. poicilipes were found only outdoors. C. pipiens was found all the year round particularly in Spring. Other species were found in Spring and Autumn, except C. antennatus which was found in Summer as well. The results were discussed on the light of work done before.
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...
Pokora, Z
1996-01-01
In the paper an attempt to define pathogenesis of changes in somatic growth of juvenile individuals of the popular freshwater snail Lymnaea stagnalis experimentally infected with parthenites of the trematode Opisthioglyphe ranae was undertaken. Significant enlargement of relative wet weight of examined accessory sex organs (albumen gland, oothecal gland, prostate, male copulatory organ) observed in infected snails permits to explain increase of their somatic growth basing on the hypothesis of disturbances in energetistic budget of the host-as a consequence of reduction by the parasite activity of the snail's reproductive system. Pathogenesis of this phenomenon has probably a complicated character, including also effect of parthenites on activity of the neurosecretory cells that control somatic growth in examined species of the snail. An argument for this standpoint is, observed in infected snails, increase of amount of neurosecretory material and RNA in cytoplasm of these cells (the light green cells of cerebral ganglia), as well as amount of the loose fraction of chromatine in their nuclei.
The effects of the herbicide diuron on survival and growth of Pacific treefrog (Pseudacris regilla),bullfrog(Rana catesbeiana), red-legged frog(Rana aurora),and African clawed frog(Xenopus laevis)embryos and tadpoles were determined in static-renewal tests. P.regilla and X.laevis...
Design and development of aqueous nanoformulations for mosquito control.
Montefuscoli, Antonela Rita; Werdin González, Jorge Omar; Palma, Santiago Daniel; Ferrero, Adriana Alicia; Fernández Band, Beatriz
2014-02-01
Microemulsions (ME) are thermodynamically stable isotropic mixtures of oil, water, and surfactant; they would also be attractive as potential insecticidal products due to the high bioviability of the active ingredient, attributable to the small sizes of the oil drops. A laboratory study was conducted in order to compare the biological effect of oil in water (o/w) geranium essential oil (EO) and geraniol MEs and emulsions, against Culex pipiens pipiens mosquito larvae. The systems were based on three nonionic surfactants (Cremophor EL, Brij 35, Tween 80). The MEs showed dispersed phase diameters in the range of 8 to 14 nm and had low PDI values (<0.2). The MEs were analyzed by TEM, indicating that they had nearly spherical morphology. The microemulsified systems based on geranium EO and those of geraniol produced a notable increase of the larvicidal activity when compared with the respectably emulsions, concluding that the biological effect is related with the diameter of the dispersed phase. The smallest drops achieved the highest larvicidal activity, being the aqueous nanoformulations based on geraniol most effective than those of geranium EO. However, geranium microemulsions are preferred due to their residual toxicological profiles. The results indicate that these novel systems could be used in integrated pest management program for the C. pipiens pipiens.
A perchlorate sensitive iodide transporter in frogs
Carr, Deborah L.; Carr, James A.; Willis, Ray E.; Pressley, Thomas A.
2008-01-01
Nucleotide sequence comparisons have identified a gene product in the genome database of African clawed frogs (Xenopus laevis) as a probable member of the solute carrier family of membrane transporters. To confirm its identity as a putative iodide transporter, we examined the function of this sequence after heterologous expression in mammalian cells. A green monkey kidney cell line transfected with the Xenopus nucleotide sequence had significantly greater 125I uptake than sham-transfected control cells. The uptake in carrier-transfected cells was significantly inhibited in the presence of perchlorate, a competitive inhibitor of mammalian Na+/iodide symporter. Tissue distributions of the sequence were also consistent with a role in iodide uptake. The mRNA encoding the carrier was found to be expressed in the thyroid gland, stomach, and kidney of tadpoles from X. laevis, as well as the bullfrog Rana catesbeiana. The ovaries of adult X. laevis also were found to express the carrier. Phylogenetic analysis suggested that the putative X. laevis iodide transporter is orthologous to vertebrate Na+-dependent iodide symporters. We conclude that the amphibian sequence encodes a protein that is indeed a functional Na+/iodide symporter in Xenopus laevis, as well as Rana catesbeiana. PMID:18275962
Boone, M.D.
2005-01-01
I reared four species of anurans (Rana sphenocephala [Southern Leopard Frog], Rana blairi [Plains Leopard Frog], Rana clamitans [Green Frog], and Bufo woodhousii [Woodhouse's Toad]) for seven to 12 months in small, outdoor terrestrial enclosures (1 x 2 m) to examine the consequences of larval competition (via density) and contaminant exposure (via the insecticide carbaryl). I added six Rana clamitans, eight Rana sphenocephala, eight Rana blairi, and 10 Bufo woodhousii to terrestrial enclosures shortly after metamorphosis and recaptured them during the following spring. All anurans from low-density ponds were significantly larger than those from high-density ponds, but these size differences did not significantly affect survival to or size at spring emergence. However, R. sphenocephala, R. blairi, and R. clamitans that survived to spring had been larger at metamorphosis on average than those that did not survive; in contrast, B. woodhousii that survived the winter were smaller at metamorphosis on average than those that did not survive. Carbaryl exposure affected mass at metamorphosis of R. clamitans and B. woodhousii that were added to enclosures, but this difference disappeared or did not increase by spring emergence. Overall, exposure to carbaryl during the larval period did not have any apparent effects on survival or growth during the terrestrial phase. In my study, anurans were able to offset small size at metamorphosis with terrestrial growth, although there was a trend of reduced overwinter survival for ranid species that metamorphosed at a smaller size. Copyright 2005 Society for the Study of Amphibians and Reptiles.
Pearl, Christopher A.; Bull, E.L.; Green, D.E.; Bowerman, Jay; Adams, Michael J.; Hyatt, A.; Wente, W.
2007-01-01
Chytridiomycosis (infection by the fungus Batrachochytrium dendrobatidis) has been associated with amphibian declines in at least four continents. We report results of disease screens from 210 pond-breeding amphibians from 37 field sites in Oregon and Washington. We detected B. dendrobatidis on 28% of sampled amphibians, and we found a?Y 1 detection of B. dendrobatidis from 43% of sites. Four of seven species tested positive for B. dendrobatidis, including the Northern Red-Legged Frog (Rana aurora), Columbia Spotted Frog (Rana luteiventris), and Oregon Spotted Frog (Rana pretiosa). We also detected B. dendrobatidis in nonnative American Bullfrogs (Rana catesbeiana) from six sites in western and central Oregon. Our study and other recently published findings suggest that B. dendrobatidis has few geographic and host taxa limitations among North American anurans. Further research on virulence, transmissibility, persistence, and interactions with other stressors is needed to assess the potential impact of B. dendrobatidis on Pacific Northwestern amphibians.
Occurrence of the amphibian pathogen Batrachochytrium dendrobatidis in the Pacific Northwest
Pearl, C.A.; Bull, E.L.; Green, D.E.; Bowerman, J.; Adams, M.J.; Hyatt, A.; Wente, W.H.
2007-01-01
Chytridiomycosis (infection by the fungus Batrachochytrium dendrobatidis) has been associated with amphibian declines in at least four continents. We report results of disease screens from 210 pond-breeding amphibians from 37 field sites in Oregon and Washington. We detected B. dendrobatidis on 28% of sampled amphibians, and we found ??? 1 detection of B. dendrobatidis from 43% of sites. Four of seven species tested positive for B. dendrobatidis, including the Northern Red-Legged Frog (Rana aurora), Columbia Spotted Frog (Rana luteiventris), and Oregon Spotted Frog (Rana pretiosa). We also detected B. dendrobatidis in nonnative American Bullfrogs (Rana catesbeiana) from six sites in western and central Oregon. Our study and other recently published findings suggest that B. dendrobatidis has few geographic and host taxa limitations among North American anurans. Further research on virulence, transmissibility, persistence, and interactions with other stressors is needed to assess the potential impact of B. dendrobatidis on Pacific Northwestern amphibians. Copyright 2007 Society for the Study of Amphibians and Reptiles.
Electrogenic active proton pump in Rana esculenta skin and its role in sodium ion transport.
Ehrenfeld, J; Garcia-Romeu, F; Harvey, B J
1985-02-01
Kinetic and electrophysiological studies were carried out in the in vitro Rana esculenta skin, bathed in dilute sodium solution, to characterize the proton pump and coupling between sodium absorption (JNa+n) and proton excretion (JH+n). JNa+n and JH+n were both dependent on transepithelial potential (psi ms); hyperpolarizing the skin decreased JNa+n and increased JH+n; depolarization produced the opposite effects. Amiloride (5 X 10(-5) M) at a clamped psi ms of +50 mV inhibited JNa+n without affecting JH+n. Variations of psi ms or pH had identical effects on JH+n. Ethoxzolamide inhibited JH+n and simultaneously increased psi ms by 15-30 mV. These changes were accompanied by depolarization of the apical membrane potential psi mc from -47 to -25 mV and an increase in apical membrane resistance of 30%; no significant effects on basolateral membrane potential (psi cs) and resistance (Rb) nor on shunt resistance (Rj) were observed. The proton pump appears to be localized at the apical membrane. The proton pump was also inhibited by deoxygenation, oligomycin, dicyclohexylcarbodiimide and vanadate (100, 78, 83 and 100% inhibition respectively). The variations of JH+n and of the measured electrical currents were significantly correlated. These findings are supportive evidence of a primary active proton pump, electrogenic and strictly linked to aerobic metabolism. The current-voltage (I-V) relation of the proton pump was obtained as the difference in the I-V curves of the apical membrane extracted before and after proton-pump inhibition by ethoxzolamide during amiloride block of sodium transport. The proton-pump current (IP) was best described by a saturable exponential function of psi mc. Maximal pump current (ImaxP) was calculated to be 200 nequiv h-1 cm-2 at a psi mc of +50 mV and the pump reversal potential ERP was -130 mV. The effect of ethoxzolamide to depolarize psi mc was dependent on the relation between psi mc and ERP. Maximal induced depolarization occurred at a
Electrogenic active proton pump in Rana esculenta skin and its role in sodium ion transport.
Ehrenfeld, J; Garcia-Romeu, F; Harvey, B J
1985-01-01
Kinetic and electrophysiological studies were carried out in the in vitro Rana esculenta skin, bathed in dilute sodium solution, to characterize the proton pump and coupling between sodium absorption (JNa+n) and proton excretion (JH+n). JNa+n and JH+n were both dependent on transepithelial potential (psi ms); hyperpolarizing the skin decreased JNa+n and increased JH+n; depolarization produced the opposite effects. Amiloride (5 X 10(-5) M) at a clamped psi ms of +50 mV inhibited JNa+n without affecting JH+n. Variations of psi ms or pH had identical effects on JH+n. Ethoxzolamide inhibited JH+n and simultaneously increased psi ms by 15-30 mV. These changes were accompanied by depolarization of the apical membrane potential psi mc from -47 to -25 mV and an increase in apical membrane resistance of 30%; no significant effects on basolateral membrane potential (psi cs) and resistance (Rb) nor on shunt resistance (Rj) were observed. The proton pump appears to be localized at the apical membrane. The proton pump was also inhibited by deoxygenation, oligomycin, dicyclohexylcarbodiimide and vanadate (100, 78, 83 and 100% inhibition respectively). The variations of JH+n and of the measured electrical currents were significantly correlated. These findings are supportive evidence of a primary active proton pump, electrogenic and strictly linked to aerobic metabolism. The current-voltage (I-V) relation of the proton pump was obtained as the difference in the I-V curves of the apical membrane extracted before and after proton-pump inhibition by ethoxzolamide during amiloride block of sodium transport. The proton-pump current (IP) was best described by a saturable exponential function of psi mc. Maximal pump current (ImaxP) was calculated to be 200 nequiv h-1 cm-2 at a psi mc of +50 mV and the pump reversal potential ERP was -130 mV. The effect of ethoxzolamide to depolarize psi mc was dependent on the relation between psi mc and ERP. Maximal induced depolarization occurred at a
Rochlin, I.; Ginsberg, H.S.; Campbell, S.R.
2009-01-01
Culex species were monitored at three proximate sites with historically different West Nile virus (WNV) activities. The site with human WNV transmission (epidemic) had the lowest abundance of the putative bridge vectors, Culex pipiens and Cx. salinarius. The site with horse cases but not human cases (epizootic) had the highest percent composition of Cx. salinarius, whereas the site with WNV-positive birds only (enzootic) had the highest Cx. pipiens abundance and percent composition. A total of 29 WNV-positive Culex pools were collected at the enzootic site, 17 at the epidemic site, and 14 at the epizootic site. Published models of human risk using Cx. pipiens and Cx. salinarius as the primary bridge vectors did not explain WNV activity at our sites. Other variables, such as additional vector species, environmental components, and socioeconomic factors, need to be examined to explain the observed patterns of WNV epidemic activity.
Mosquito species involved in the circulation of West Nile and Usutu viruses in Italy.
Mancini, Giuseppe; Montarsi, Fabrizio; Calzolari, Mattia; Capelli, Gioia; Dottori, Michele; Ravagnan, Silvia; Lelli, Davide; Chiari, Mario; Santilli, Adriana; Quaglia, Michela; Quaglia, Michela; Federici, Valentina; Monaco, Federica; Goffredo, Maria; Savini, Giovanni
2017-06-30
Usutu (USUV) and West Nile (WNV) are mosquito-borne Flavivirus emerged in Italy in 1996 and 1998, respectively, and reappeared 10 years later. The aim of this work is to review the Italian mosquito species found positive for WNV and USUV between 2008 and 2014. Moreover, the role of mosquitoes in promoting the overwintering of these viruses is discussed, as a result of the mosquito collections performed in Molise region between September 2010 and April 2011. Overall 99,000 mosquitoes were collected: 337 and 457 mosquito pools tested positive by real time reverse transcriptase polymerase chain reaction (real time RT-PCR) for WNV and USUV, respectively. West Nile virus was detected in pools of Culex pipiens s.l. (329), Ochlerotatus caspius (4), Culex modestus (2), and Culex spp. (2). Positive USUV pools were from Cx. pipiens s.l. (435), Aedes albopictus (12), Oc. caspius (5), Culex spp. (2), Anopheles maculipennis s.l. (1), Culiseta annulata (1), and Ochlerotatus detritus (1). In Molise region, 1,694 mosquitoes were collected, and USUV was identi ed in Cx. pipiens s.l., Cs. annulata, and Oc. detritus pools. This paper shows that Cx. pipiens s.l. is the mosquito species most involved in the WNV and USUV circulation in Italy, although other species would also support the spread of both the viruses during Winter.
Pancharatna, M; Saidapur, S K
1984-03-01
The effect of homoplastic pituitary pars distalis homogenate (PDH), PMSG, and HCG on the postovulatory follicles/corpora lutea (CL) of the frog Rana cyanophlyctis was studied to elucidate the factors regulating the life span of the luteal cells. Ovulation and spawning was induced in hypophysectomized frogs using PDH. Starting from Day 1 of spawning 1/2 PDH, 50 IU PMSG, or 50 IU HCG was injected daily for 3 days. In the saline-injected control frogs, the granulosa lutein cells regressed markedly on Day 2 with a steady progressive increase in the pycnosis of their nuclei. The sudanophilic lipid droplets of the luteal cells were fine on Day 1 but became coarser and reduced in number on subsequent days. Histochemically, the luteal cell 3 beta-HSDH and G-6-PDH also decreased drastically by Day 2. In PDH-treated frogs the granulosa lutein cells were healthy on all 4 days of the experiment. The nuclear diameter of the luteal cells increased progressively due to PDH. The pycnosis of the luteal cells was limited to 7.6% on Day 4 due to PDH as opposed to 68% seen in the controls. Histochemically, 3 beta-HSDH and G-6-PDH activities remained much higher than in the controls with abundant sudanophilic lipids (both fine and coarse) in the luteal cells of PDH treated frogs even on Day 4. PMSG treatment also maintained the granulosa lutein cells beyond their normal life span but the luteotrophic effect was less than that of PDH. HCG was least effective. The present studies suggest that the structural integrity of CL in the frog can be extended beyond the normal life span by injecting PDH or PMSG.
Effect of temperature on residual force enhancement in single skeletal muscle fibers.
Lee, Eun-Jeong; Herzog, Walter
2008-08-28
It is well accepted that the steady-state isometric force following active stretching of a muscle is greater than the steady-state isometric force obtained in a purely isometric contraction at the same length. This property of skeletal muscle has been called residual force enhancement (FE). Despite decades of research the mechanisms responsible for FE have remained largely unknown. Based on previous studies showing increases in FE in fibers in which cross-bridges were biased towards weakly bound states, we hypothesized that FE might be associated with a stretch-induced facilitation of transitioning from weakly to strongly bound cross-bridges. In order to test this hypothesis, single fibers (n=11) from the lumbrical muscles of frog (Rana pipiens) were used to determine FE at temperatures of 7 and 20 degrees C. At the cold temperature, cross-bridges are biased towards weakly bound states, therefore we expected FE to be greater at 7 degrees C compared to 20 degrees C. The average FE was significantly greater at 7 degrees C (11.5+/-1.1%) than at 20 degrees C (7.8+/-1.0%), as expected. The enhancement of force/stiffness was also significantly greater at the low (13.3+/-1.4%) compared to the high temperature (5.6+/-1.7%), indicating an increased conversion from weakly to strongly bound cross-bridges at the low temperature. We conclude from the results of this study that muscle preparations that are biased towards weakly bound cross-bridge states show increased FE for given stretch conditions, thereby supporting the idea that FE might be caused, in part, by a stretch-induced facilitation of the conversion of weakly to strongly bound cross-bridges.
Euliss, Ned H.; Mushet, David M.
2004-01-01
We compared the macroinvertebrate and amphibian communities of 12 excavated and 12 natural wetlands in western North Dakota, USA, to assess the effects of artificially lengthened hydroperiods on the biotic communities of wetlands in this semi-arid region. Excavated wetlands were much deeper and captured greater volumes of water than natural wetlands. Most excavated wetlands maintained water throughout the study period (May to October 1999), whereas most of the natural wetlands were dry by June. Excavated wetlands were largely unvegetated or contained submergent and deep-marsh plant species. The natural wetlands had two well-defined vegetative zones populated by plant species typical of wet meadows and shallow marshes. Excavated wetlands had a richer aquatic macroinvertebrate community that included several predatory taxa not found in natural wetlands. Taxa adapted to the short hydroperiods of seasonal wetlands were largely absent from excavated wetlands. The amphibian community of natural and excavated wetlands included the boreal chorus frog (Pseudacris maculata), northern leopard frog (Rana pipiens), plains spadefoot (Scaphiopus bombifrons), Woodhouse's toad (Bufo woodhousii woodhousii), and tiger salamander (Ambystoma tigrinum). The plains spadefoot occurred only in natural wetlands while tiger salamanders occurred in all 12 excavated wetlands and only one natural wetland. Boreal chorus frogs and northern leopard frogs were present in both wetland types; however, they successfully reproduced only in wetlands lacking tiger salamanders. Artificially extending the hydroperiod of wetlands by excavation has greatly influenced the composition of native biotic communities adapted to the naturally short hydroperiods of wetlands in this semi-arid region. The compositional change of the biotic communities can be related to hydrological changes and biotic interactions, especially predation related to excavation.
Neurotropic effects of venoms and other factors that promote prey acquisition.
Gennaro, Joseph Francis; Hall, Hollis Pace; Casey, Ernest Raymond; Hayes, William Kent
2007-09-01
Mammals envenomed by either the Eastern diamondback rattlesnake (Crotalus adamanteus) or the cottonmouth (Agkistrodon piscivorus piscivorus) exhibit an immediate but transitory pupillar contraction, a parasympathomimetic effect mediated through the ciliary ganglion that can be prevented by a retrobulbar injection of anesthetic. The venom of the cottonmouth injected into the lymph spaces of the frog (Rana pipiens) produces an immediate and total collapse of the lung sacs. Applied locally to the surface, it produces a constriction that eventually collapses the entire sac. Tests of venoms and toxins from both anterior and posterior parts of the venom apparatus indicate that the lung-collapsing moiety originates in the accessory, not the main portion of the venom gland. This is the first example of a functional specialization within the whole structure. It seems that this factor is elaborated primarily in snakes that prey upon frogs, although insufficient data are available from this study to confirm this. In both reptile species, the predatory strike is accompanied by an immediate effect, perhaps mediated by the parasympathetic nervous system, designed to incapacitate the prey and facilitate capture. These effects cannot now be attributed to neurotoxins because the effect of the former is transitory (and not lethal) and neither has been purified sufficiently to determine potency or structure. Both take part in securing, but not killing, the prey, and both directly oppose the sympathetic nervous system "fright-fight/flight" response. Evidence is presented to support the possibility that known epigenetic mechanisms are capable of effecting heritable changes in gene expression that could allow for the development of factors that facilitate prey acquisition and promote rapid adaptation to environmental change. (c) 2007 Wiley-Liss, Inc.
Unlu, Isik; Farajollahi, Ary; Strickman, Daniel; Fonseca, Dina M.
2013-01-01
Our ultimate objective is to design cost-effective control strategies for Aedes albopictus, the Asian tiger mosquito, an important urban nuisance and disease vector that expanded worldwide during the last 40 years. We conducted mosquito larval surveys from May through October 2009 in the City of Trenton, New Jersey, USA, while performing intensive monthly source-reduction campaigns that involved removing, emptying, or treating all accessible containers with larvicides and pupicides. We examined patterns of occurrence of Ae. albopictus and Culex pipiens, another urban mosquito, among different container types by comparing observed and expected number of positive containers of each type. Expected use was based on the relative frequency of each container type in the environment. Aedes albopictus larvae and pupae were found significantly more often than expected in medium volumes of water in buckets and plant saucers but were rarely collected in small volumes of water found in trash items such as discarded cups and cans. They were also absent from large volumes of water such as in abandoned swimming pools and catch basins, although we consistently collected Cx. pipiens from those habitats. The frequency of Ae. albopictus in tires indicated rapid and extensive use of these ubiquitous urban containers. Standard larval-based indices did not correlate with adult catches in BG-Sentinel traps, but when based only on Ae. albopictus key containers (buckets, plant saucers, equipment with pockets of water, and tires) they did. Although we found that only 1.2% of the 20,039 water-holding containers examined contained immature Ae. albopictus (5.3% if only key containers were counted), adult populations were still above nuisance action thresholds six times during the 2009 mosquito season. We conclude that in urban New Jersey, effective source reduction for Ae. albopictus control will require scrupulous and repeated cleaning or treatment of everyday use containers and extensive
Toma, Luciano; Catalani, Monica; Catalano, Antonella; Goffredo, Maria; Romi, Roberto; Di Luca, Marco
2016-08-03
This study reports the mosquito collections conducted in July-August 2013 in Castiglione del Lago (Umbria Region, Italy) during the entomological surveillance within the West Nile national control program. The mosquito collections showed the noteworthy occurrence of Anopheles hyrcanus s.l. (n = 156; 35.8% of the whole sample), a relatively rare mosquito species in Italy, with molecular analyses con rming that these mosquitoes belong to An. hyrcanus s.s. The same catching site was characterized by a relevant richness in mosquito species, in particular the following were found: Anopheles maculipennis s.l. (n = 146; 33.5% composed by 80% Anopheles melanoon, 13.3% Anopheles maculipennis s.s. and 6.7% Anopheles atroparvus), Culex pipiens s.l. (n = 116; 26.6% composed by 90% Cx. p. pipiens, 5% Cx. p. molestus and 5% Cx. p. pipiens/molestus), Culiseta annulata (n = 10; 2.3%), Aedes albopictus (n = 3; 0.7%), and Anopheles claviger (n = 5; 100%).
Veeranagoudar, Dheeraj K; Shanbhag, Bhagyashri A; Saidapur, Srinivas K
2004-06-01
The ability of bronze frog Rana temporalis tadpoles (pure or mixed parental lines) to assess the profitability of food habitats and distribute themselves accordingly was tested experimentally using a rectangular choice tank with a non- continuous input design. Food (boiled spinach) was placed at two opposite ends of the choice tank in a desired ratio (1 : 1, 1 : 2 or 1 : 4) to create habitat A and B. The tadpoles in Gosner stage 28-33, pre-starved for 24 h, were introduced in an open ended mesh cylinder placed in the center of the choice tank, held for 4 min (for acclimation) and then released to allow free movement and habitat selection. The number of tadpoles foraging at each habitat was recorded at 10, 15, 20, 25 and 30 min time intervals. The actual suitability, Si (the food available in a habitat after colonization of tadpoles) of each habitat was obtained from the equation Si = Bi - fi (di) where Bi is basic suitability (amount of food provided at each habitat before release of tadpoles), fi is the rate of depletion of food (lowering effect) with introduction of each tadpole, and di is the density of tadpoles in habitat i. The expected number of tadpoles at each habitat was derived from the actual suitability. With no food in the choice tank, movement of the tadpoles in the test arena was random indicating no bias towards any end of the choice tank or the procedure. In tests with a 1 : 1 food ratio, the observed ratio of tadpoles (11.71 : 12.28) was comparable with the expected 12 : 12 ratio. The observed number of tadpoles in the habitats with a 1 : 2 food ratio was 8.71 : 15.29 and 7.87 : 16.13 for pure and mixed parental lines respectively. In both cases, the observed ratios were close to the expected values (7 : 17). Likewise, in experiments with a 1 : 4 food ratio, the observed number of tadpoles in the two habitats (10.78 : 37.22) did not differ significantly from the expected ratio of 7 : 41. In all tests, the number of R. temporalis tadpoles matched
Lee, Dong Chan; Ahn, Young-Joon
2013-01-01
The toxicity of Pinus densiflora (red pine) hydrodistillate, its 19 constituents and 28 structurally related compounds against early third-instar larvae of Aedes albopictus (Ae. albopictus), Aedes aegypti (Ae. aegypti) and Culex pipiens palles (Cx. p. pallens) was examined using direct-contact bioassays. The efficacy of active compounds was further evaluated in semi-field bioassays using field-collected larval Cx. p. pallens. Results were compared with those of two synthetic larvicides, temephos and fenthion. In laboratory bioassays, Pinus densiflora hydrodistillate was found to have 24 h LC50 values of 20.33, 21.01 and 22.36 mg/L against larval Ae. albopictus, Ae. aegypti and Cx. p. pallens respectively. Among the identified compounds, thymol, δ-3-carene and (+)-limonene exhibited the highest toxicity against all three mosquito species. These active compounds were found to be nearly equally effective in field trials as well. In vitro bioassays were conducted to examine the acetylcholinesterase (AChE) inhibitory activity of 10 selected compounds. Results showed that there is a noticeable correlation between larvicidal activity and AChE inhibitory activity. In light of global efforts to find alternatives for currently used insecticides against disease vector mosquitoes, Pinus densiflora hydrodistillate and its constituents merit further research as potential mosquito larvicides. PMID:26464387
West Nile Virus lineage-2 in Culex specimens from Iran.
Shahhosseini, Nariman; Chinikar, Sadegh; Moosa-Kazemi, Seyed Hassan; Sedaghat, Mohammad Mehdi; Kayedi, Mohammad Hassan; Lühken, Renke; Schmidt-Chanasit, Jonas
2017-10-01
Screening of mosquitoes for viruses is an important forecasting tool for emerging and re-emerging arboviruses. Iran has been known to harbour medically important arboviruses such as West Nile virus (WNV) and dengue virus (DENV) based on seroepidemiological data. However, there are no data about the potential mosquito vectors for arboviruses in Iran. This study was performed to provide mosquito and arbovirus data from Iran. A total of 32 317 mosquitos were collected at 16 sites in five provinces of Iran in 2015 and 2016. RT-PCR for detection of flaviviruses was performed. The PCR amplicons were sequenced, and 109 WNV sequences, including one obtained in this study, were used for phylogenetic analyses. The 32 317 mosquito specimens belonging to 25 species were morphologically distinguished and distributed into 1222 pools. Culex pipiens s.l. comprised 56.429%. One mosquito pool (0.08%), containing 46 unfed Cx. pipiens pipiens form pipiens (Cpp) captured in August 2015, was positive for flavivirus RNA. Subsequent sequencing and phylogenetic analyses revealed that the detected Iranian WNV strain belongs to lineage 2 and clusters with a strain recently detected in humans. No flaviviruses other than WNV were detected in the mosquito pools. Cpp could be a vector for WNV in Iran. Our findings indicate recent circulation of WNV lineage-2 strain in Iran and provide a solid base for more targeted arbovirus surveillance programs. © 2017 John Wiley & Sons Ltd.
Straus, Christian; Samara, Ziyad; Fiamma, Marie-Noëlle; Bautin, Nathalie; Ranohavimparany, Anja; Le Coz, Patrick; Golmard, Jean-Louis; Darré, Pierre; Zelter, Marc; Poon, Chi-Sang; Similowski, Thomas
2011-05-01
Human ventilation at rest exhibits mathematical chaos-like complexity that can be described as long-term unpredictability mediated (in whole or in part) by some low-dimensional nonlinear deterministic process. Although various physiological and pathological situations can affect respiratory complexity, the underlying mechanisms remain incompletely elucidated. If such chaos-like complexity is an intrinsic property of central respiratory generators, it should appear or increase when these structures mature or are stimulated. To test this hypothesis, we employed the isolated tadpole brainstem model [Rana (Pelophylax) esculenta] and recorded the neural respiratory output (buccal and lung rhythms) of pre- (n = 8) and postmetamorphic tadpoles (n = 8), at physiologic (7.8) and acidic pH (7.4). We analyzed the root mean square of the cranial nerve V or VII neurograms. Development and acidosis had no effect on buccal period. Lung frequency increased with development (P < 0.0001). It also increased with acidosis, but in postmetamorphic tadpoles only (P < 0.05). The noise-titration technique evidenced low-dimensional nonlinearities in all the postmetamorphic brainstems, at both pH. Chaos-like complexity, assessed through the noise limit, increased from pH 7.8 to pH 7.4 (P < 0.01). In contrast, linear models best fitted the ventilatory rhythm in all but one of the premetamorphic preparations at pH 7.8 (P < 0.005 vs. postmetamorphic) and in four at pH 7.4 (not significant vs. postmetamorphic). Therefore, in a lower vertebrate model, the brainstem respiratory central rhythm generator accounts for ventilatory chaos-like complexity, especially in the postmetamorphic stage and at low pH. According to the ventilatory generators homology theory, this may also be the case in mammals.
Lenhardt, Patrick P; Brühl, Carsten A; Leeb, Christoph; Theissinger, Kathrin
2017-01-01
Amphibian populations have been declining globally over the past decades. The intensification of agriculture, habitat loss, fragmentation of populations and toxic substances in the environment are considered as driving factors for this decline. Today, about 50% of the area of Germany is used for agriculture and is inhabited by a diverse variety of 20 amphibian species. Of these, 19 are exhibiting declining populations. Due to the protection status of native amphibian species, it is important to evaluate the effect of land use and associated stressors (such as road mortality and pesticide toxicity) on the genetic population structure of amphibians in agricultural landscapes. We investigated the effects of viniculture on the genetic differentiation of European common frog ( Rana temporaria ) populations in Southern Palatinate (Germany). We analyzed microsatellite data of ten loci from ten breeding pond populations located within viniculture landscape and in the adjacent forest block and compared these results with a previously developed landscape permeability model. We tested for significant correlation of genetic population differentiation and landscape elements, including land use as well as roads and their associated traffic intensity, to explain the genetic structure in the study area. Genetic differentiation among forest populations was significantly lower (median pairwise F ST = 0.0041 at 5.39 km to 0.0159 at 9.40 km distance) than between viniculture populations (median pairwise F ST = 0.0215 at 2.34 km to 0.0987 at 2.39 km distance). Our analyses rejected isolation by distance based on roads and associated traffic intensity as the sole explanation of the genetic differentiation and suggest that the viniculture landscape has to be considered as a limiting barrier for R. temporaria migration, partially confirming the isolation of breeding ponds predicted by the landscape permeability model. Therefore, arable land may act as a sink habitat, inhibiting
NASA Astrophysics Data System (ADS)
Trawinski, P. R.; Mackay, D. S.
2009-03-01
The objective of this study is to quantify and model spatial dependence in mosquito vector populations and develop predictions for unsampled locations using geostatistics. Mosquito control program trap sites are often located too far apart to detect spatial dependence but the results show that integration of spatial data over time for Cx. pipiens-restuans and according to meteorological conditions for Ae. vexans enables spatial analysis of sparse sample data. This study shows that mosquito abundance is spatially correlated and that spatial dependence differs between Cx. pipiens-restuans and Ae. vexans mosquitoes.
Rapid assessment of mosquitoes and arbovirus activity after floods in southeastern Kansas, 2007.
Harrison, Bruce A; Whitt, Parker B; Roberts, Lesa F; Lehman, Jennifer A; Lindsey, Nicole P; Nasci, Roger S; Hansen, Gail R
2009-09-01
A rapid assessment was conducted in July-August 2007 to determine the impact of heavy rains and early summer floods on the mosquitoes and arbovirus activity in 4 southeastern Kansas counties. During 10 days and nights of collections using different types and styles of mosquito traps, a total of 10,512 adult female mosquitoes representing 29 species were collected, including a new species record for Kansas (Psorophora mathesoni). High numbers of Aedes albopictus were collected. Over 4,000 specimens of 4 Culex species in 235 species-specific pools were tested for the presence of West Nile, St. Louis, and western equine encephalitis viruses. Thirty pools representing 3 Culex species were positive for West Nile virus (WNV). No other arboviruses were detected in the samples. Infection rates of WNV in Culex pipiens complex in 2 counties (10.7/1,000 to 22.6/1,000) and in Culex salinarius in 1 county (6.0/1,000) were sufficiently high to increase the risk of transmission to humans. The infection rate of WNV in Culex erraticus was 1.9/1,000 in one county. Two focal hot spots of intense WNV transmission were identified in Montgomery and Wilson counties, where infection rates in Cx. pipiens complex were 26/ 1,000 and 19.9/1,000, respectively. Despite confirmed evidence of WNV activity in the area, there was no increase in human cases of arboviral disease documented in the 4 counties for the remainder of 2007.
Melvin, Steven D; Trudeau, Vance L
2012-08-01
Naphthenic acids (NA) have been identified as harmful environmental contaminants that influence survival, growth and development of wildlife. Amphibian larvae are particularly susceptible to waterborne contaminants, but little information exists regarding exposure of amphibian embryos or tadpoles to NA. Our results demonstrate that embryos of Lithobates pipiens and Silurana tropicalis exposed to 2-4 mg/l of a commercial NA blend suffer significant reductions (32% and 25%, respectively) in growth and development upon hatching. Increased incidences of deformities were observed in exposed individuals of both species, but were only significant in L. pipiens. Embryos suffered 100% mortality following exposure to 6 mg/l NAs, and narcosis at lower concentrations. LC50 estimates were 4.10 mg/l for 72-h exposure in L. pipiens and 4.13, 3.51, and 2.95 mg/l for 24-, 48-, and 72-h exposure in S. tropicalis. These data suggest that exposure to NAs at environmentally realized concentrations may negatively affect tadpole populations. Copyright © 2012 Elsevier Ltd. All rights reserved.
Effects of lambda-cyhalothrin on mosquito larvae and predatory aquatic insects.
Lawler, Sharon P; Dritz, Deborah A; Christiansen, Julie A; Cornel, Anthony J
2007-03-01
Agricultural insecticides can affect mosquito production in rice fields by controlling mosquitoes, disrupting biological control or contributing to selection of insecticide resistance. The duration of insecticidal activity of the pyrethroid lambda-cyhalothrin was quantified on predatory insects in rice fields and on three kinds of mosquito larva: a pyrethroid-susceptible strain of Culex tarsalis Coquillet, a pyrethroid-resistant strain of Cx pipiens L. (sensu lato) and non-resistant Cx pipiens s.l. Lambda-cyhalothrin killed most caged, susceptible mosquitoes for up to 21 days. It killed fewer resistant Cx pipiens s.l., but suppressed their survival for over a week. Lambda-cyhalothrin suppressed field populations of predatory insects through day 29. Agricultural use of lambda-cyhalothrin can provide incidental mosquito control. However, the pyrethroid persisted in sediment and gradually decreased in activity, which could contribute to selection of pyrethroid-resistant mosquitoes. Because caged mosquitoes showed good survival before predators recovered, disruption of biological control is possible. It is therefore advisable for growers and mosquito control agencies to communicate about pesticide use. Copyright 2006 Society of Chemical Industry.
2008-10-31
of the Apalachicola River drainage. Although this proposed division in classification appears to be generally accepted by the herpetological community...breeding in small forest ponds. Herpetological Review 33(4):275-280. Carle, F. L. and M. R. Strub. 1978. A new method for estimating population size...gopher frogs (Rana capito) and southern leopard frogs (Rana sphenocephala). Journal of Herpetology 42: 97-103. Grevstad, F.S. 2005. Simulating
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
Samara, Ziyad; Fiamma, Marie-Noëlle; Bautin, Nathalie; Ranohavimparany, Anja; Le Coz, Patrick; Golmard, Jean-Louis; Darré, Pierre; Zelter, Marc; Poon, Chi-Sang; Similowski, Thomas
2011-01-01
Human ventilation at rest exhibits mathematical chaos-like complexity that can be described as long-term unpredictability mediated (in whole or in part) by some low-dimensional nonlinear deterministic process. Although various physiological and pathological situations can affect respiratory complexity, the underlying mechanisms remain incompletely elucidated. If such chaos-like complexity is an intrinsic property of central respiratory generators, it should appear or increase when these structures mature or are stimulated. To test this hypothesis, we employed the isolated tadpole brainstem model [Rana (Pelophylax) esculenta] and recorded the neural respiratory output (buccal and lung rhythms) of pre- (n = 8) and postmetamorphic tadpoles (n = 8), at physiologic (7.8) and acidic pH (7.4). We analyzed the root mean square of the cranial nerve V or VII neurograms. Development and acidosis had no effect on buccal period. Lung frequency increased with development (P < 0.0001). It also increased with acidosis, but in postmetamorphic tadpoles only (P < 0.05). The noise-titration technique evidenced low-dimensional nonlinearities in all the postmetamorphic brainstems, at both pH. Chaos-like complexity, assessed through the noise limit, increased from pH 7.8 to pH 7.4 (P < 0.01). In contrast, linear models best fitted the ventilatory rhythm in all but one of the premetamorphic preparations at pH 7.8 (P < 0.005 vs. postmetamorphic) and in four at pH 7.4 (not significant vs. postmetamorphic). Therefore, in a lower vertebrate model, the brainstem respiratory central rhythm generator accounts for ventilatory chaos-like complexity, especially in the postmetamorphic stage and at low pH. According to the ventilatory generators homology theory, this may also be the case in mammals. PMID:21325645
Becnel, J J; Johnson, M A
1998-12-01
Nine species of mosquitoes and several species of non-target aquatic organisms were tested for susceptibility to the mernaithid nematode, Strelkovimermis spiculatus. All species of Anopheles, Aedes, Culex, and Toxorhynchites exposed to S. spiculatus were susceptible. Of the nine mosquito species tested, C. pipiens quinquefasciatus had the greatest tolerance to initial invasion and the highest percent infection of those that survived. High levels of infection were also achieved with Aedes taeniorhynchus and A. albopictus, but these mosquitoes were significantly less tolerant to parasitism than C. pipiens quinquefasciatus. Strelkovimermis spiculatus did not infect or develop in any of the non-target hosts tested.
Kirby, Matthew J; West, Philippa; Green, Clare; Jasseh, Momodou; Lindsay, Steve W
2008-01-01
Background Screening doors, windows and eaves of houses should reduce house entry by eusynanthropic insects, including the common African house mosquito Culex pipiens quinquefasciatus and other culicines. In the pre-intervention year of a randomized controlled trial investigating the protective effects of house screening against mosquito house entry, a multi-factorial risk factor analysis study was used to identify factors influencing house entry by culicines of nuisance biting and medical importance. These factors were house location, architecture, human occupancy and their mosquito control activities, and the number and type of domestic animals within the compound. Results 40,407 culicines were caught; the dominant species were Culex thalassius, Cx. pipiens s.l., Mansonia africanus, M. uniformis and Aedes aegypti. There were four times more Cx. pipiens s.l. in Farafenni town (geometric mean/trap/night = 8.1, 95% confidence intervals, CIs = 7.2–9.1) than in surrounding villages (2.1, 1.9–2.3), but over five times more other culicines in the villages (25.1, 22.1–28.7) than in town (4.6, 4.2–5.2). The presence of Cx. pipiens s.l. was reduced in both settings if the house had closed eaves (odds ratios, OR town = 0.62, 95% CIs = 0.49–0.77; OR village = 0.49, 0.33–0.73), but increased per additional person in the trapping room (OR town = 1.16, 1.09–1.24; OR village = 1.10, 1.02–1.18). In the town only, Cx. pipiens s.l. numbers were reduced if houses had a thatched roof (OR = 0.70, 0.51–0.96), for each additional cow tethered near the house (OR = 0.73, 0.65–0.82) and with increasing distance from a pit latrine (OR = 0.97, 0.95–0.99). In the villages a reduction in Cx. pipiens s.l. numbers correlated with increased horses in the compound (OR = 0.90, 0.82–0.99). The presence of all other culicines was reduced in houses with closed eaves (both locations), with horses tethered outside (village only) and with increasing room height (town only), but
Assessing the toxicity and teratogenicity of pond water in north-central Minnesota to amphibians.
Bridges, Christine; Little, Edward; Gardiner, David; Petty, James; Huckins, James
2004-01-01
Incidence of amphibian deformities have increased in recent years, especially in the northern region of the United States. While many factors have been proposed as being responsible for generating deformities (e.g., contaminants, ultraviolet radiation [UV], parasites), no single cause has been definitively established. To determine whether waterborne chemicals are responsible for amphibian deformities in ponds in north-central Minnesota, we deployed semipermeable membrane devices (SPMDs) in an impacted and a reference site to accumulate lipophilic contaminants. We then exposed native tadpoles (northern leopard frogs; Rana pipiens) to the SPMD extracts combined with two agricultural pesticides (atrazine, carbaryl) at two levels of UV radiation. UV radiation alone caused a slight increase in hatching success and tadpole growth rate. Deformity rate among hatchlings was high following exposure to SPMD extracts from the reference site in the absence of UV, suggesting that chemicals present at this site are broken down by UV to less harmful forms, or become less bioavailable. Conversely, impacted site SPMD extracts caused hatchling deformities only in the presence of UV, suggesting that UV potentiates the teratogenicity of the compounds present there. Impacted site SPMD extracts significantly increased the number of bony triangles among metamorphs, a common deformity observed at this site. The incidence of skin webbings increased significantly with SPMD extracts from both sites as well as with our pesticide control containing atrazine and carbaryl alone. Higher deformity rates among tadpoles reared in the presence of UV radiation and SPMD extracts from sites where deformities are common indicates a chemical compound (or compounds) in the water at this site may be causing the deformities. It is important to examine the effects of chemical stressors in the presence of other natural stressors (e.g., UV radiation) to gain a better understanding of how multiple stressors work
2018-01-01
The objective was to better understand how a series compliance alters contraction kinetics and power output of muscle to enhance the work done on a load. A mathematical model was created in which a gravitational point load was connected via a linear spring to a muscle (based on the contractile properties of the sartorius of leopard frogs, Rana pipiens). The model explored the effects of load mass, tendon compliance, and delay between onset of contraction and release of the load (catch) on lift height and power output as measures of performance. Series compliance resulted in increased lift height over a relatively narrow range of compliances, and the effect was quite modest without an imposed catch mechanism unless the load was unrealistically small. Peak power of the muscle-tendon complex could be augmented up to four times that produced with a muscle alone, however, lift height was not predicted by peak power. Rather, lift height was improved as a result of the compliance synchronizing the time courses of muscle force and shortening velocity, in particular by stabilizing shortening velocity such that muscle power was sustained rather than rising and immediately falling. With a catch mechanism, enhanced performance resulted largely from energy storage in the compliance during the period of catch, rather than increased time for muscle activation before movement commenced. However, series compliance introduced a trade-off between work done before versus after release of the catch. Thus, the ability of tendons to enhance locomotor performance (i.e. increase the work done by muscle) appears dependent not only on their established role in storing energy and increasing power, but also on their ability to modulate the kinetics of muscle contraction such that power is sustained over more of the contraction, and maximizing the balance of work done before versus after release of a catch. PMID:29370246
What we know and don't know about amphibian declines in the West
Corn, Paul Stephen
1994-01-01
The problem of declining amphibian species is thought to be particularly acute in western North America, but there are many gaps in our knowledge. Although several declines have been well-documented, other declines are anecdotal or hypothesized. Most documented declines are of ranid frogs or toads (Bufo). Species from montane habitats and those occurring in California have been best studied. Status of many desert species is unknown. Habitat destruction and introduced predators are the most common threats to amphibian populations. Some declines may represent natural variation in population size. Causes have not been determined for several cases where common species have declined over large areas. There are important considerations for ecosystem management, whether changes in amphibian populations are natural or caused by human activities. Causes for declines must be known so that management can be prescribed (or proscribed) to eliminate or minimize these causes. The natural variability of amphibian population numbers and the complexity of metapopulation structure emphasize the necessity of considering multiple temporal and spatial scales in ecosystem management. The decline of amphibian species throughout the world has received considerable recent attention (e.g., Blaustein and Wake 1990, Griffiths and Beebee 1992, Yoffe 1992). Much of this attention derives from a workshop held in February, 1990 on declining amphibians sponsored by the National Research Council Board (NRC) on Biology in Irvine, California (Barinaga 1990, Borchelt 1990). Because of media attention in the aftermath of this conference, it is a popular perception that amphibian declines are a new phenomenon that herpetologists have been slow to recognize (Griffiths and Beebee 1992, Quammen 1993). However, concern about amphibian populations in the United States dates back over 20 years. Beginning in the 1960s, a large, well-documented decline of northern leopard frogs (Rana pipiens) occurred in the
Antiviral effect of ranpirnase against Ebola virus.
Hodge, Thomas; Draper, Ken; Brasel, Trevor; Freiberg, Alexander; Squiquera, Luis; Sidransky, David; Sulley, Jamie; Taxman, Debra J
2016-08-01
The recent epidemic of Ebola has intensified the need for the development of novel antiviral therapeutics that prolong and improve survival against deadly viral diseases. We sought to determine whether ranpirnase, an endoribonuclease from Rana pipiens with a demonstrated human safety profile in phase III oncology trials, can reduce titers of Ebola virus (EBOV) in infected cells, protect mice against mouse-adapted EBOV challenge, and reduce virus levels in infected mice. Our results demonstrate that 0.50 μg/ml ranpirnase is potently effective at reducing EBOV Zaire Kikwit infection in cultured Vero E6 cells (Selectivity Index 47.8-70.2). In a prophylactic study, a single intravenous dose of 0.1 mg/kg ranpirnase protected 70% of mice from progressive infection. Additionally, in a post-exposure prophylactic study, 100% of female mice survived infection after intraperitoneal administration of 0.1 mg/kg ranpirnase for ten days beginning 1 h post challenge. Most of the male counterparts were sacrificed due to weight loss by Study Day 8 or 9; however, the Clinical Activity/Behavior scores of these mice remained low and no significant microscopic pathologies could be detected in the kidneys, livers or spleens. Furthermore, live virus could not be detected in the sera of ranpirnase-treated mice by Study Day 8 or in the kidneys, livers or spleens by Study Day 12, and viral RNA levels declined exponentially by Study Day 12. Because ranpirnase is exceptionally stable and has a long track record of safe intravenous administration to humans, this drug provides a promising new candidate for clinical consideration in the treatment of Ebola virus disease alone or in combination with other therapeutics. Copyright © 2016 Elsevier B.V. All rights reserved.
New insights into the passive force enhancement in skeletal muscles.
Lee, Eun-Jeong; Joumaa, Venus; Herzog, Walter
2007-01-01
The steady-state isometric force following active stretching of a muscle is always greater than the steady-state isometric force obtained in a purely isometric contraction at the same length. This phenomenon has been termed "residual force enhancement" and it is associated with an active and a passive component. The origin of these components remains a matter of scientific debate. The purpose of this work was to test the hypothesis that the passive component of the residual force enhancement is caused by a passive structural element. In order to achieve this purpose, single fibers (n=6) from the lumbrical muscles of frog (Rana pipiens) were isolated and attached to a force transducer and a motor that could produce computer-controlled length changes. The passive force enhancement was assessed for three experimental conditions: in a normal Ringer's solution, and after the addition of 5 and 15mM 2,3-butanedione monoxime (BDM) which inhibits force production in a dose-dependent manner. If our hypothesis was correct, one would expect the passive force enhancement to be unaffected following BDM application. However, we found that increasing concentrations of BDM decreased the isometric forces, increased the normalized residual force enhancement, and most importantly for this study, increased the passive force enhancement. Furthermore, BDM decreased the rate of force relaxation after deactivation following active stretching of fibers, passive stretching in the Ringer's and BDM conditions produced the same passive force-sarcomere length relationship, and passive force enhancement required activation and force production. These results led to the conclusion that the passive force enhancement cannot be caused by a structural component exclusively as had been assumed up to date, but must be associated, directly or indirectly, with cross-bridge attachments upon activation and the associated active force.
The Intensity Of The 2.7nm Reflection As A Constraint For Models Of Myosin Docking To Actin
DOE Office of Scientific and Technical Information (OSTI.GOV)
Reconditi, Massimo; Irving, Tom C.; IIT)
2009-03-16
Previous workers have proposed high resolution models for the docking of the myosin heads on actin on the basis of combined crystallographic and electron microscopy data (Mendelson and Morris, 1997 PNAS 94:8533; Holmes et al. 2003 Nature 425:423). We have used data from small angle X-ray fiber diffraction from living muscle to check the predictions of these models. Whole sartorius muscles from Rana pipiens were mounted in a chamber containing Ringer's solution at 10 C and at rest length at the BioCAT beamline (18 ID, Advanced Photon Source, Argonne, IL-U.S.A.). The muscles were activated by electrical stimulation and the forcemore » was recorded with a muscle lever system type 300B (Aurora Scientific). X-ray patterns were collected with 1s total exposures at rest and during isometric contraction out to 0.5 nm{sup -1} in reciprocal space, as the higher angle reflections are expected to be more sensitive to the arrangement of myosin heads on actin. We observed that during isometric contraction the meridional reflection originating from the 2.73nm repeat of the actin monomers along the actin filament increases its intensity by a factor 2.1 {+-} 0.2 relative to rest. Among the models tested, Holmes et al. fits the data when the actin filament is decorated with 30-40% the total available myosin heads, a fraction similar to that estimated with fast single fiber mechanics by Piazzesi et al. (2007, Cell 131:784). However, when the mismatch between the periodicities of actin and myosin filaments is taken into account, none of the models can reproduce the fiber diffraction data. We suggest that the fiber diffraction data should be used as a further constraint on new high resolution models for the docking of the myosin heads on actin.« less
Green, D.E.; Converse, K.A.; Majumdar, S.K.; Huffman, J.E.; Brenner, F.J.; Panah, A.I.
2005-01-01
(Rana pipiens). Other interesting diseases could have been presented, such as a wart-like virus infection of Japanese newts and a group of protistan parasites, referred to as Dermocystidium and Dermomycoides, in European frogs and toads. The reader is referred to Green (2001) for a review of these diseases. Amphibians have a rich diversity of helminthic parasites (Poynton and Whitaker, 2001). In general, most cestodes, trematodes and nematodes of amphibians are innocuous and not linked to specific clinical signs ('symptoms') or mortalities. An important major exception to this generalization is the trematode, Ribeiroia, which has been linked to numerous and bizarre malformations of frogs, toads and salamanders (Johnson et al., 1999, Johnson et al., 2001, Schotthoefer et al., 2003). Two genera of trematodal parasites are discussed in this chapter: Ribeiroia because they cause malformations and Clinostomum because they are large and produce visible lumps in the skin. For a review of amphibian helminths, the reader is referred to the text by Flynn (1973).
Zhan, Aibin; Li, Cheng; Fu, Jinzhong
2009-04-09
Amphibians in general are poor dispersers and highly philopatric, and landscape features often have important impacts on their population genetic structure and dispersal patterns. Numerous studies have suggested that genetic differentiation among amphibian populations are particularly pronounced for populations separated by mountain ridges. The Tsinling Mountain range of northern China is a major mountain chain that forms the boundary between the Oriental and Palearctic zoogeographic realms. We studied the population structure of the Chinese wood frog (Rana chensinensis) to test whether the Tsinling Mountains and the nearby Daba Mountains impose major barriers to gene flow. Using 13 polymorphic microsatellite DNA loci, 523 individuals from 12 breeding sites with geographical distances ranging from 2.6 to 422.8 kilometers were examined. Substantial genetic diversity was detected at all sites with an average of 25.5 alleles per locus and an expected heterozygosity ranging from 0.504 to 0.855, and two peripheral populations revealed significantly lower genetic diversity than the central populations. In addition, the genetic differentiation among the central populations was statistically significant, with pairwise FST values ranging from 0.0175 to 0.1625 with an average of 0.0878. Furthermore, hierarchical AMOVA analysis attributed most genetic variation to the within-population component, and the between-population variation can largely be explained by isolation-by-distance. None of the putative barriers detected from genetic data coincided with the location of the Tsinling Mountains. The Tsinling and Daba Mountains revealed no significant impact on the population genetic structure of R. chensinensis. High population connectivity and extensive juvenile dispersal may account for the significant, but moderate differentiation between populations. Chinese wood frogs are able to use streams as breeding sites at high elevations, which may significantly contribute to the
Adult repellency and larvicidal activity of five plant essential oils against mosquitoes.
Zhu, Junwei; Zeng, Xiaopeng; Yanma; Liu, Ting; Qian, Kuen; Han, Yuhua; Xue, Suqin; Tucker, Brad; Schultz, Gretchen; Coats, Joel; Rowley, Wayne; Zhang, Aijun
2006-09-01
The larvicidal activity and repellency of 5 plant essential oils--thyme oil, catnip oil, amyris oil, eucalyptus oil, and cinnamon oil--were tested against 3 mosquito species: Aedes albopictus, Ae. aegypti, and Culex pipiens pallens. Larvicidal activity of these essentials oils was evaluated in the laboratory against 4th instars of each of the 3 mosquito species, and amyris oil demonstrated the greatest inhibitory effect with LC50 values in 24 h of 58 microg/ml (LC90 = 72 microg/ml) for Ae. aegypti, 78 microg/ml (LC90 = 130 microg/ml) for Ae. albopictus, and 77 microg/ml (LC90 = 123 microg/ml) for Cx. p. pallens. The topical repellency of these selected essential oils and deet against laboratory-reared female blood-starved Ae. albopictus was examined. Catnip oil seemed to be the most effective and provided 6-h protection at both concentrations tested (23 and 468 microg/ cm2). Thyme oil had the highest effectiveness in repelling this species, but the repellency duration was only 2 h. The applications using these natural product essential oils in mosquito control are discussed.
Resistance Level of Mosquito Species (Diptera: Culicidae) from Shandong Province, China.
Liu, Hong-Mei; Yang, Pei-Pei; Cheng, Peng; Wang, Hai-Fang; Liu, Li-Juan; Huang, Xiaodan; Zhao, Yu-Qiang; Wang, Huai-Wei; Zhang, Chong-Xing; Gong, Mao-Qing
2015-01-01
This study describes the aquatic habitats, species composition, and the insecticide resistance level of the mosquito Culex pipiens pallens in Shandong Province, China. A cross-sectional survey of mosquito larval habitats was conducted from May to November 2014 to determine the species composition and larval abundance. Larvae were collected using the standard dipping technique, and a total of four habitat types were sampled. The fourth instar larvae of Cx. pipiens pallens collected in each habitat type were tested for resistance to five insecticides according to a WHO bioassay. A total of 7,281 mosquito larvae were collected, of which 399 (5.48%) were categorized as Anopheles mosquito larvae (An. sinensis), 6636 (91.14%) as culicine larvae (Cx. pipiens pallens, Cx. tritaeniorhynchus, Cx. halifaxii, and Cx. bitaeniorhynchus), 213 (2.93%) as Armigeres larvae, and 33 (0.45%) as Aedes larvae (Aedes albopictus). In addition, a total of 1,149 mosquito pupae were collected. Culex larvae were distributed in all habitats investigated. Tukeys HSD analysis showed that roadside drainages were the most productive habitat type for Culex larvae. Armigeres species were found only in drains, Aedes only in water tanks, and Anopheles in water that was comparatively clear and rich in emergent plants. Bioassay showed that the maximum resistance level of Cx. pipiens pallens was to deltamethrin, while it was lowest to plifenate. The productivity of various mosquitoes in different habitat types is very heterogeneous. It is particularly important to modify human activity and the environment to achieve effective mosquito vector control. For effective larval control, the type of habitat should be considered, and the most productive habitat type should be given priority in mosquito abatement programs.
Vu, Maria; Weiler, Bradley; Trudeau, Vance L
2017-12-01
Gonadotropin-releasing hormone (GnRH) stimulates luteinizing hormone release to control ovulation and spermiation in vertebrates. Dopamine (DA) has a clear inhibitory role in the control of reproduction in numerous teleosts, and emerging evidence suggests that similar mechanisms may exist in amphibians. The interactions between GnRH and DA on spawning success and pituitary gene expression in the Northern leopard frog (Lithobates pipiens) were therefore investigated. Frogs were injected during the natural breeding season with a GnRH agonist [GnRH-A; (Des-Gly 10 , D-Ala 6 , Pro-NHEt 9 )-LHRH; 0.1μg/g and 0.4μg/g] alone and in combination with the dopamine receptor D2 antagonist metoclopramide (MET; 5μg/g and 10μg/g). Injected animals were allowed to breed in outdoor mesocosms. Time to amplexus and oviposition were assessed, and egg mass release, incidences of amplexus, egg mass weight, total egg numbers and fertilization rates were measured. To examine gene expression, female pituitaries were sampled at 12, 24 and 36h following injection of GnRH-A (0.4μg/g) alone and in combination with MET (10μg/g). The mRNA levels of the genes lhb, fshb, gpha, drd2 and gnrhr1 were measured using quantitative real-time PCR. Data were analyzed by a two-way ANOVA. Both GnRH-A doses increased amplexus, oviposition and fertilization alone. Co-injection of MET with GnRH-A did not further enhance spawning success. Injection of GnRH-A alone time-dependently increased expression of lhb, fshb, gpha and gnrhr1. The major effect of MET alone was to decrease expression of drd2. Importantly, the stimulatory effects of GnRH-A on lhb, gpha and gnrhr1 were potentiated by the co-injection of MET at 36h. At this time, expression of fshb was increased only in animals injected with both GnRH-A and MET. Spawning success was primarily driven by the actions of GnRH-A. The hypothesized inhibitory action of DA was supported by pituitary gene expression analysis. The results from this study provide a
Bahgat, Iman Mohamed
2013-08-01
The distribution and monthly abundance of mosquito larvae in released water, drainage canals and sewage water tanks in Ismailia governorate were investigated. The results obtained indicated the presence of five culicine (Culex. pipiens, Cx. pusillus, Cx. perexiguus, Cx. theleri and Ochlerotatus. caspius) and two anopheline (Anopheles. multicolor and An. pharoensis) mosquito species. Significantly higher larval density was recorded in sewage water (n= 5534; 46.08%) as compared with released water (n = 2903; 24.17%) and drainage water (n= 3573; 29.75%). Culex pipiens was the most dominant mosquito species in the three habitats. The effects of environmental parameters including pH, biological and chemical oxygen demands, day time water temperature, plant growth, salinity, total organic matter and concentrations of heavy metals on larval population density were investigated. The positive correlations observed between heavy metals concentrations in the three habitats suggested relative uniformity of the sources of metal pollutants. Culex pipiens larvae demonstrated high tolerance to elevated levels of heavy metals in sewage water and compensatory effects of high nutrient levels generally associated with sewage or domestic waste. High densities of culicine larvae were accompanied by low density of anopheline larvae. This was attributed to water chemistry & competitive interactions.
Hematophagous insects as vectors for frog trypanosomes.
Ramos, B; Urdaneta-Morales, S
1977-12-01
Experimental infections of three hematophagous arthropods (Rhodnius prolixus, Aedes aegypti, and Culex pipiens) with a trypanosome of the Trypanosoma rotatorium complex found in the frogs Hyla crepitans and Leptodactylus insularum revealed that A. aegypti is a good host for the flagellate; the course of development in the intestinal tract of the mosquito is described from 15 minutes to 168 hours. C. pipiens showed only low intestinal infections and R. prolixus did not permit development of the parasite. It is postulated that, in addition to the transmission of T. rotatorium by leeches, batrachophilic mosquitoes may transmit the parasite to frogs of more terrestrial habits by being ingested by these anurans.
The response of Rana muscosa, the mountain yellow-legged frog, to short distance translocations.
K. R. Matthews
2003-01-01
ABSTRACT.âTo determine the response of Mountain Yellow-Legged Frogs to short distance translocations, I placed transmitters on 20 adult frogs and moved them short distances from 144â630 m and monitored their responses for up to 30 days. Of the 20 translocated frogs, seven frogs returned to their original capture site, four frogs moved in the direction of their capture...
Yaghoobi-Ershadi, M R; Doosti, S; Schaffner, F; Moosa-Kazemi, S H; Akbarzadeh, K; Yaghoobi-Ershadi, N
2017-05-01
Beside numerous extensive studies on Anophelinae mosquitoes of Iran, little is known on Aedes species in the country and existing reports are dispersed. The objective of this study was to identify adults of Culicinae species occurring in the Sistan and Baluchestan Province, southeast of Iran, during 2012-2014. Mosquito collections were carried out three times (May-June, September, October-November) in four counties by Centers for Diseases Control (CDC) light traps and human landing catches outdoors and under bed nets baited with carbon dioxide. These trapping were carried out two consecutive nights during the field studies. Several mosquito collections were also conducted with aspirator and pyrethrum spray space catches during the day. A total of 1885 mosquitoes were collected, belonging to 10 species of genus Culex including Cx. pipiens Complex, Cx. laticinctus, Cx. sinaiticus, and Cx. tritaeniorhynchus, one species of the genus Culiseta, Cs. longiareolata, and five species of the genus Aedes, Ae. caspius, Ae. vexans, Ae. detritus, Ae. albopictus, and Ae. unilineatus. Ae. vexans was the dominant species in the area (77.7%). During the study, seven Ae. unilineatus were collected in two villages near the city of Chabahar located in a coastal area; this is the first record for Iran and identification was confirmed by Cytochrome oxidase (COI) sequences analysis. Confirmation of the presence of Ae. unilineatus in the country raises the number of species of the genus Aedes to 12. The detection of this species reveals its probable establishment in the southeast of the country, which has implications for public health such as dengue and Zika infections and requires active entomological surveillance and implementation of adapted vector control measures in the area.
Usutu virus persistence and West Nile virus inactivity in the Emilia-Romagna region (Italy) in 2011.
Calzolari, Mattia; Bonilauri, Paolo; Bellini, Romeo; Albieri, Alessandro; Defilippo, Francesco; Tamba, Marco; Tassinari, Massimo; Gelati, Antonio; Cordioli, Paolo; Angelini, Paola; Dottori, Michele
2013-01-01
The circulation of West Nile virus and Usutu virus was detected in the Emilia-Romagna region in 2008 and 2009. To evaluate the extent of circulation of both viruses, environmental surveillance, based on bird and mosquito testing, was conducted in 2008 and gradually improved over the years. In February-March 2009-2011, 5,993 hibernating mosquitoes were manually sampled, out of which 80.1% were Culex pipiens; none tested positive for the viruses. From 2008 to 2011, 946,213 mosquitoes, sampled between May and October, were tested; 86.5% were Cx. pipiens. West Nile virus was detected in 32 Cx. pipiens pools, and Usutu virus was detected in 229 mosquito pools (217 Cx. pipiens, 10 Aedes albopictus, one Anopheles maculipennis s.l., and one Aedes caspius). From 2009 to 2011, of 4,546 birds collected, 42 tested positive for West Nile virus and 48 for Usutu virus. West Nile virus and Usutu virus showed different patterns of activity during the 2008-2011 surveillance period. West Nile virus was detected in 2008, 2009, and 2010, but not in 2011. Usutu virus, however, was continuously active throughout 2009, 2010, and 2011. The data strongly suggest that both viruses overwinter in the surveyed area rather than being continually reintroduced every season. The lack of hibernating mosquitoes testing positive for the viruses and the presence of positive birds sampled early in the season support the hypothesis that the viruses overwinter in birds rather than in mosquitoes. Herd immunity in key bird species could explain the decline of West Nile virus observed in 2011, while the persistence of Usutu virus may be explained by not yet identified reservoirs. Reported results are comparable with a peri-Mediterranean circulation of the West Nile virus lineage 1 related strain, which became undetectable in the environment after two to three years of obvious circulation.
Desser, S S
2001-02-01
During May 1997, specimens of 7 species of anurans, that included 5 Phrynohyas venulosa Laurenti, 5 Rana forreri Boulenger, 7 Rana vaillanti Brucchi, 6 Eleutherodactylus fitzingeri Schimdt, 4 Smilisca baudinii Duméril and Bibron, 1 Leptodactylus melanonotus, and 3 Bufo marinus Linneaus, from the Guanacaste Conservation Area, Costa Rica were examined for blood parasites. Their hematozoan fauna included intraerythrocytic and intraleukocytic icosahedral viruses, a rickettsia (Aegyptianella sp.), 2 species of Hepatozoon, Lankesterella minima, 2 unknown species of apicomplexans, 9 morphologically distinct types of trypanosomes, and 2 species of microfilariae. Rana vaillanti, the most aquatic species of frog, harbored the most species of parasites. Recent evidence indicates that morphological changes in the highly pleomorphic trypanosomes of anurans from different geographical regions have not kept pace with biochemical (isozyme) and molecular (DNA sequence) changes. Describing new species based solely on bloodstream trypomastigotes is discouraged. Additional criteria described herein should be applied when naming new species of anuran trypanosomes.
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