Reproductive Biology, Sperm storage, and Sexual Maturity of Thamnodynastes strigatus (Serpentes: Dipsadidae).

Life history strategies determine and influence many aspects of species fitness. In this study, we describe the reproductive biology - reproductive cycle, sperm storage, and sexual maturity - of Thamnodynastes strigatus in South Brazil. We analyzed 49 individuals (25 males and 24 females) from herpetological collections. The reproductive cycle of males and females was described considering the morpho-anatomical and histological changes in the testes, ductus deferens, and kidney, as well in the ovary and oviduct. The age at the onset of sexual maturity was determined by skeletochronology of the caudal vertebra. The reproductive cycle is seasonal semi-synchronous and most individuals have a reproductive peak in spring and summer. The seasonal biennial reproductive cycle and viviparity are two phylogenetically conserved characters in Tachymenini snakes. Thamnodynastes Strigatus females store sperm in the utero-vaginal junction furrows during autumn. There were no differences between the ages of sexual maturity of males (4-11y) and females (4-12y). Females reach sexual maturity at larger body sizes, and this may confer an adaptive advantage due to a higher fecundity potential. Herein, we confirmed the previously described seasonal biennial reproductive cycle of T. strigatus through histological analysis.

Reproductive biology, sperm storage, and sexual maturity of Thamnodynastes strigatus (Serpentes: Dipsadidae)

Life history strategies determine and influence many aspects of species fitness. In this study, we describe the reproductive biology - reproductive cycle, sperm storage, and sexual maturity - of Thamnodynastes strigatus in South Brazil. We analyzed 49 individuals (25 males and 24 females) from herpetological collections. The reproductive cycle of males and females was described considering the morpho-anatomical and histological changes in the testes, ductus deferens, and kidney, as well in the ovary and oviduct. The age at the onset of sexual maturity was determined by skeletochronology of the caudal vertebra. The reproductive cycle is seasonal semi-synchronous and most individuals have a reproductive peak in spring and summer. The seasonal biennial reproductive cycle and viviparity are two phylogenetically conserved characters in Tachymenini snakes. Thamnodynastes Strigatus females store sperm in the utero-vaginal junction furrows during autumn. There were no differences between the ages of sexual maturity of males (4-11y) and females (4-12y). Females reach sexual maturity at larger body sizes, and this may confer an adaptive advantage due to a higher fecundity potential. Herein, we confirmed the previously described seasonal biennial reproductive cycle of T. strigatus through histological analysis.

First record of growth rings for 11 native subtropical anuran species of South America.

Skeletochronology is the most accurate method to estimate a population age structure. The methodology is based on the analysis of secondary bone tissue in order to count growth rings. This study aimed to provide initial data, allowing researchers to further work out in the age of individuals and populations, sampling evidence of the presence of growth rings in 11 native species (representing nine families) of a subtropical region of southern Brazil. Four bone samples of each specimen were used to perform the skeletochronological analysis: the penultimate phalanges of the 3rd and 4th fingers, the humerus, and the femur. The presence of growth rings was confirmed in the periosteal layer of the bones of all analyzed species. In comparison with phalanges, growth rings of humeri and femora are more irregular and less distinguishable. This is the first record of growth rings to the native species herein analised. The skeletochronology was proved to be an effective tool in determining the age of anuran amphibians from a subtropical region, since this environment presents well defined climatic seasonality.

Morphological variation and sexual dimorphism in two sympatric dipsadine snakes from Southern Brazil

Snakes have high morphological variation, including between sexes. Most of these variations are evidenced only by linear measurements, which are generally restricted to size traits. Using traditional and geometric morphometrics methods we analyze how body size attributes and head size and shape varies between sexes and species of two sympatric Tachymenini snakes, Thamnodynastes strigatus and Tomodon dorsatum. We took body size measurements from 87 specimens of T. strigatus and 83 specimens of T. dorsatum. We also extracted head shape and size of 146 specimens of T. strigatus and 156 specimens of T. dorsatum. Our results showed sexual differences in some body size attributes and in head shape and size. Females of both species had larger SVL, BM, HW, BW and heads than in males. But, T. strigatus is not sexually dimorphic in head size. Females of the T. strigatus also exhibited an enlargement of the parietal region compared to the males, while T. dorsatum females exhibited a wider snout than males. These sexual differences could be a result of fecundity selection and may also indicate that females consume larger preys than conspecific males. At the species level, T. strigatus is larger in body and head size than T. dorsatum. The head shape of T. strigatus is characterized by a more elongated head with larger frontal and prefrontal scales and slender snout compared to T. dorsatum. Shape differences between these two snakes probably evolved in association with divergences in habitat use e.g. terrestrial and arboreal and diet specialization e.g. piscivory, and malacophagy.

In vitro Safety and Efficacy of Lavender Essential Oil (Lamiales: Lamiaceae) as an Insecticide Against Houseflies (Diptera: Muscidae) and Blowflies (Diptera: Calliphoridae).

Essential oils are considered an alternative for replacement of conventional insecticides that have provoked an increasing number of resistant species and damages to health. The objective of this work was to investigate the insecticidal activity of Lavandula dentata L. (Lamiales: Lamiaceae) oil against the species Musca domestica L. (Diptera: Muscidae) and Chrysomya albiceps Wiedemann (Diptera: Calliphoridae). Extraction of the essential oil from the aerial parts of the plant was carried out using hydro-distillation. Its principal compounds (1,8-cineol, camphor, and linalool oxide) were identified. Insecticidal activity was determined by evaluating adulticidal effect using topical application methods and exposure to impregnated paper; larvicidal effect was determined using immersion method. The essential oil presented toxicity in M. domestica and C. albiceps adults; the lethal concentration values (LC50) in the superficial application method were respectively 3.13 ± 0.64 and 1.39 ± 0.19% live weight (l/v). Toxicity was also found in the oil impregnated paper exposure test; the LC50 results for M. domestica and C. albiceps were respectively 4.15 ± 0.64 and 5.14 ± 0.81%. Larvicidal effect was observed on third stage M. domestica larvae when exposed to an oil concentration of 2.5% (m/v). In addition, the oil was evaluated for cytotoxicity, mutagenicity, and genotoxicity in human cells, and the in vitro safety of this oil in human cells was verified. Thus, L. dentata oil presented insecticidal activity in M. domestica and C. albiceps and can be used as an alternative for control of these dipterans.

Sunlight-induced genotoxicity and damage in keratin structures decrease tadpole performance.

The increased incidence of solar ultraviolet (UV) radiation, an environmental genotoxic agent, due to ozone depletion or deforestation may help to explain the enigmatic decline of amphibian populations in specific localities. In this work, we evaluated the importance of DNA repair performed by photolyases to maintain the performance of treefrog tadpoles after acute and chronic treatments with environmental-simulated doses of solar UVB and UVA radiation. Immediately after UV treatments, tadpoles were exposed to a visible light source to activate photolyases or kept in dark containers. The biological effects of UV treatments were evaluated through morphological, histological, locomotor and survival analyzes of Boana pulchella tadpoles (Anura: Hylidae). The results indicate that tadpole body weight suffered influence after both UVB and UVA treatments, although the body length was bit affected. The locomotor performance of UVB-exposed tadpoles was significantly reduced. In addition, UVB radiation induced a severe impact on tadpole skin, as well as on keratinized structures of mouth (tooth rows and jaw), indicating that these should be important effects of solar UV radiation in the reduction of tadpole performance. Furthermore, photolyases activation was fundamental for the maintenance of tadpole performance after chronic UVB exposures, but it was relatively inefficient after acute exposures to UVB, but not to UVA radiation. Therefore, this work demonstrates how the UV-induced genotoxicity and structural alterations in the skin and oral apparatus affect tadpole performance and survival.

Reproductive biology of Philodryas patagoniensis (Snakes: Dipsadidae) in south Brazil: female reproductive cycle

In this study, we describe the female reproductive cycle of Philodryas patagoniensis in south Brazil, which was described through morpho-anatomical and histological analyses. The peak of secondary vitellogenesis occurred during winter-spring (July-December), ovulation in spring (October-December), mating and fertilization in spring-summer (October-February), oviposition in spring-autumn (October-May) and births from late spring to autumn (December-July). The diameter of vitellogenic follicles/eggs was larger in winter-spring than in other seasons. The diameter of the shell glands was also larger in winter-spring. In spite of the clear reproductive peak, gonads only showed reduced activity in the autumn. Therefore, at the individual level, females have a discontinuous cyclical reproduction; in the populational level, the reproductive cycle is seasonal semisynchronous. We support the hypothesis that P.patagoniensis have the ability to produce multiple clutches with long-term stored sperm. Sexual dimorphism in body size was evident, and females are significantly larger and heavier than males. Larger females were able to produce follicles and eggs in larger amount and size. The maternal body size was positively related to the reproductive effort and fecundity. To conclude, we deliberated about the proximal and distal causes that influence the reproductive traits and patterns of P.patagoniensis.

Humic acid of commercial origin causes changes in gill morphology of silver catfish Rhamdia quelen exposed to acidic water: Response of silver catfish gills to low pH and humic acid.

The effect of dissolved organic carbon (DOC) against the burden of low pH was assessed in silver catfish Rhamdia quelen through the gills. A commercial humic acid (HA) was used as the source of DOC at 0 (control), 10, 25, and 50 mg/l. For each HA concentration, two pH levels were tested: 6.5 (control) and 5.5. After 40 days of exposure, the gills were removed and morphological variables were analyzed through light and scanning electronic microscopy. The low water pH caused a reduction in the length of filaments, number of lamellae, and gill respiratory surface area (GRSA) and an increase in chloride cells (CCs) number in the filament epithelium. When HA was added to the test water, GRSA increased and CCs proliferated in lamellae, suggesting ionic and respiratory disturbances. Scanning electronic microscopy revealed that CC morphometric variables, that is, apical area, fractional area, and density, were undisturbed by the pH reduction, but increased in the presence of HA. This study indicates that the commercial HA did not protect silver catfish against low pH stress. Instead, it caused changes that may affect vital processes such as ion regulation and ventilation and, consequently, reduce growth.

Growth of silver catfish ( Rhamdia quelen ) exposed to acidic pH at different humic acid levels / Crescimento de jundiás (Rhamdia quelen) expostos ao pH ácido em diferentes níveis de ácido húmico

ABSTRACT: Humic substances are organic compounds that possess high molecular weight and are formed by the decomposition of plant materials. Humic substances comprise humic acids, fulvic acids and humin. Humic acids (HA) have shown to be able to provide some protection to biological membranes of fish in water with low pH, replacing the protective action of Ca2+. Thus, this studyaimed to observe growth of silver catfish ( Rhamdia quelen ) juveniles exposed to pHs 5.5 and 6.5 and to different levels of HA: 0, 10, 25 and 50mg L-1. Results showed that, irrespective of pH, HA was highly detrimental to silver catfish growth, since the higher the concentration of HA, the lower the weight gain and feed intake, resulting in lower biomass and lower specific growth rate of juveniles. Hence, HA is extremely damaging to the performance of silver catfish juveniles in the tested concentrations.

RESUMO: Substâncias húmicas são compostos orgânicos que possuem elevado peso molecular e são formadas pela decomposição da matéria orgânica de plantas. As substâncias húmicas compreendem ácidos húmicos, ácidos fúlvicos e humina. Os ácidos húmicos (AH) têm mostrado serem capazes de proporcionar proteção parcial das membranas biológicas de peixes em águas com baixo pH, substituindo a ação protetora do Ca2+. Assim, este estudo teve como objetivo observar o crescimento de juvenis de jundiá ( Rhamdia quelen ) expostos a pH's 5,5 e 6,5 com diferentes níveis de AH: 0, 10, 25 e 50mg L-1. Os resultados demostram que, independentemente do pH, o AH foi altamente prejudicial para o crescimento do jundiá, uma vez que, quanto maior era a concentração de AH, menor era o ganho de peso e a ingestão de alimentos, resultando em menor biomassa e taxa de crescimento específico inferior dos juvenis. Assim, os AH são extremamente prejudiciais para o desempenho de juvenis de jundiá nas concentrações testadas.

Growth of silver catfish (Rhamdia quelen) exposed to acidic pH at different humic acid levels / Crescimento de jundiás (Rhamdia quelen) expostos ao pH ácido em diferentes níveis de ácido húmico

Humic substances are organic compounds that possess high molecular weight and are formed by the decomposition of plant materials. Humic substances comprise humic acids, fulvic acids and humin. Humic acids (HA) have shown to be able to provide some protection to biological membranes of fish in water with low pH, replacing the protective action of Ca2+. Thus, this studyaimed to observe growth of silver catfish ( Rhamdia quelen ) juveniles exposed to pHs 5.5 and 6.5 and to different levels of HA: 0, 10, 25 and 50mg L-1. Results showed that, irrespective of pH, HA was highly detrimental to silver catfish growth, since the higher the concentration of HA, the lower the weight gain and feed intake, resulting in lower biomass and lower specific growth rate of juveniles. Hence, HA is extremely damaging to the performance of silver catfish juveniles in the tested concentrations.(AU)

Substâncias húmicas são compostos orgânicos que possuem elevado peso molecular e são formadas pela decomposição da matéria orgânica de plantas. As substâncias húmicas compreendem ácidos húmicos, ácidos fúlvicos e humina. Os ácidos húmicos (AH) têm mostrado serem capazes de proporcionar proteção parcial das membranas biológicas de peixes em águas com baixo pH, substituindo a ação protetora do Ca2+. Assim, este estudo teve como objetivo observar o crescimento de juvenis de jundiá ( Rhamdia quelen ) expostos a pH's 5,5 e 6,5 com diferentes níveis de AH: 0, 10, 25 e 50mg L-1. Os resultados demostram que, independentemente do pH, o AH foi altamente prejudicial para o crescimento do jundiá, uma vez que, quanto maior era a concentração de AH, menor era o ganho de peso e a ingestão de alimentos, resultando em menor biomassa e taxa de crescimento específico inferior dos juvenis. Assim, os AH são extremamente prejudiciais para o desempenho de juvenis de jundiá nas concentrações testadas.(AU)

Effect of humic acid on survival, ionoregulation and hematology of the silver catfish, Rhamdia quelen (Siluriformes: Heptapteridae), exposed to different pHs

This study evaluates whether humic acid (HA; Aldrich) protects the silver catfish, Rhamdia quelen (Quoy & Gaimard, 1824), against exposure to acidic pH. Survival, levels of Na+, Cl- and K+ plasma, hematocrit, hemoglobin and erythrocyte morphometry were measured. Fish were exposed to 0, 10, 25 and 50 mg L 1 HA at four pH levels: 3.8, 4.0, 4.2 and 7.0 up to 96 hours. None of the fish exposed to pH 3.8 survived for 96 hours into the experiment, and survival of fish subjected to pH 4.0 decreased when HA concentration increased. Plasma Na+ levels decreased when pH was acidic, with no influence of HA, while Cl- levels declined at low pH with increased HA concentration. The levels of K+ at pH 4.0 and 4.2 increased without HA. Hematocrit and hemoglobin augmented under the effect of HA. At pH 4.0 and 4.2, erythrocytes of fish not exposed to HA were smaller, an effect that was partially offset by the presence of HA, since the values at pH 7.0 were higher. Although HA showed some positive effects changes in hematological and plasma K+ª in silver catfish caused by exposure to acidic pH, the overall findings suggest that HA does not protect this species against acidic pH because it increased mortality and Cl- loss at pH 4.0.

Effect of humic acid on survival, ionoregulation and hematology of the silver catfish, Rhamdia quelen (Siluriformes: Heptapteridae), exposed to different pHs

This study evaluates whether humic acid (HA; Aldrich) protects the silver catfish, Rhamdia quelen (Quoy & Gaimard, 1824), against exposure to acidic pH. Survival, levels of Na+, Cl- and K+ plasma, hematocrit, hemoglobin and erythrocyte morphometry were measured. Fish were exposed to 0, 10, 25 and 50 mg L 1 HA at four pH levels: 3.8, 4.0, 4.2 and 7.0 up to 96 hours. None of the fish exposed to pH 3.8 survived for 96 hours into the experiment, and survival of fish subjected to pH 4.0 decreased when HA concentration increased. Plasma Na+ levels decreased when pH was acidic, with no influence of HA, while Cl- levels declined at low pH with increased HA concentration. The levels of K+ at pH 4.0 and 4.2 increased without HA. Hematocrit and hemoglobin augmented under the effect of HA. At pH 4.0 and 4.2, erythrocytes of fish not exposed to HA were smaller, an effect that was partially offset by the presence of HA, since the values at pH 7.0 were higher. Although HA showed some positive effects changes in hematological and plasma K+ª in silver catfish caused by exposure to acidic pH, the overall findings suggest that HA does not protect this species against acidic pH because it increased mortality and Cl- loss at pH 4.0.(AU)

Anesthetics products for Hungarian carp fingerlings ("Cyprinus carpio") / Produtos anestésicos para juvenis de carpa húngara ("Cyprinus carpio")

In the aquaculture procedures, the fishes are easily stressed in differetn rearing phases. So, is important the use of anesthetics for to prevent physical injuries and promote metabolism decrease, resulting in improved welfare for fish. The aim of this study was analyze the anesthetical induction of Hungarian carp fingerlings (Cyprinus Carpio) displayed of the different dosages of eugenol (20, 30, 40 and 50mg/L), ketamine (300, 600, 900 and 1200mg/L), xylazine (800, 1100, 2000mg/L) and the ketamine/xylazine association (300/300, 600/600, 900/900 and 1200/1200mg/L). Each dosage was tested for ten fish, individually subjected to the anesthetic bath with container containing 1 liter of water. All experimental procedures were the same as in the tests. After, it was analyzed the anesthetic time (TA) and the recovery time (TR). The anesthetics eugenol, ketamine and the ketamine/xylazine association allowed the deep anesthesia, but xilazine didnt allow it. Among the anesthetics tested, eugenol at 50mg/L demonstrated better anesthetic induction and recovery time for carp juveniles.

Nos procedimentos em piscicultura, os peixes são facilmente estressados nas diferentes etapas de criação. Assim, é importante o uso de anestésicos para prevenir injúrias físicas e promover a redução do metabolismo, resultando em melhor bem-estar aos peixes. O objetivo deste estudo foi analisar a indução anestésica em juvenis de carpa húngara (Cyprinus Carpio) expostos a diferentes doses de eugenol (20; 30; 40 e 50mg/L), ketamina (300; 600; 900 e 1200mg/L), xilazina (800; 1100; 2000mg/L) e da associação ketamina/xilazina (300/300; 600/600; 900/900 e 1200/1200mg/L). Cada dosagem foi testada em grupos de dez peixes escolhidos ao acaso (n=10), submetidos individualmente ao banho com anestésico em recipiente contendo 1 litro de água. Todos os procedimentos experimentais foram iguais nos ensaios. Após, foram analisados os tempos de anestesia (TA) e de recuperação (TR). Os anestésicos eugenol, ketamina e a associação ketamina/xilazina permitiram que os juvenis de carpa húngara atingissem a anestesia profunda. Entretanto, os dois últimos produtos testados causaram mortalidade em algumas doses. A xilazina não promoveu anestesia em nenhuma das doses testadas. Dentre os anestésicos testados, o eugenol na dose 50mg/L, demonstrou indução anestésica e tempo de recuperação satisfatório para juvenis de carpa.

Anesthetics products for Hungarian carp fingerlings ("Cyprinus carpio") / Produtos anestésicos para juvenis de carpa húngara ("Cyprinus carpio")

In the aquaculture procedures, the fishes are easily stressed in differetn rearing phases. So, is important the use of anesthetics for to prevent physical injuries and promote metabolism decrease, resulting in improved welfare for fish. The aim of this study was analyze the anesthetical induction of Hungarian carp fingerlings (Cyprinus Carpio) displayed of the different dosages of eugenol (20, 30, 40 and 50mg/L), ketamine (300, 600, 900 and 1200mg/L), xylazine (800, 1100, 2000mg/L) and the ketamine/xylazine association (300/300, 600/600, 900/900 and 1200/1200mg/L). Each dosage was tested for ten fish, individually subjected to the anesthetic bath with container containing 1 liter of water. All experimental procedures were the same as in the tests. After, it was analyzed the anesthetic time (TA) and the recovery time (TR). The anesthetics eugenol, ketamine and the ketamine/xylazine association allowed the deep anesthesia, but xilazine didnt allow it. Among the anesthetics tested, eugenol at 50mg/L demonstrated better anesthetic induction and recovery time for carp juveniles.(AU)

Nos procedimentos em piscicultura, os peixes são facilmente estressados nas diferentes etapas de criação. Assim, é importante o uso de anestésicos para prevenir injúrias físicas e promover a redução do metabolismo, resultando em melhor bem-estar aos peixes. O objetivo deste estudo foi analisar a indução anestésica em juvenis de carpa húngara (Cyprinus Carpio) expostos a diferentes doses de eugenol (20; 30; 40 e 50mg/L), ketamina (300; 600; 900 e 1200mg/L), xilazina (800; 1100; 2000mg/L) e da associação ketamina/xilazina (300/300; 600/600; 900/900 e 1200/1200mg/L). Cada dosagem foi testada em grupos de dez peixes escolhidos ao acaso (n=10), submetidos individualmente ao banho com anestésico em recipiente contendo 1 litro de água. Todos os procedimentos experimentais foram iguais nos ensaios. Após, foram analisados os tempos de anestesia (TA) e de recuperação (TR). Os anestésicos eugenol, ketamina e a associação ketamina/xilazina permitiram que os juvenis de carpa húngara atingissem a anestesia profunda. Entretanto, os dois últimos produtos testados causaram mortalidade em algumas doses. A xilazina não promoveu anestesia em nenhuma das doses testadas. Dentre os anestésicos testados, o eugenol na dose 50mg/L, demonstrou indução anestésica e tempo de recuperação satisfatório para juvenis de carpa.(AU)

Produtos anestésicos para juvenis de carpa húngara ("Cyprinus carpio")

In the aquaculture procedures, the fishes are easily stressed in differetn rearing phases. So, is important the use of anesthetics for to prevent physical injuries and promote metabolism decrease, resulting in improved welfare for fish. The aim of this study was analyze the anesthetical induction of Hungarian carp fingerlings (Cyprinus Carpio) displayed of the different dosages of eugenol (20, 30, 40 and 50mg/L), ketamine (300, 600, 900 and 1200mg/L), xylazine (800, 1100, 2000mg/L) and the ketamine/xylazine association (300/300, 600/600, 900/900 and 1200/1200mg/L). Each dosage was tested for ten fish, individually subjected to the anesthetic bath with container containing 1 liter of water. All experimental procedures were the same as in the tests. After, it was analyzed the anesthetic time (TA) and the recovery time (TR). The anesthetics eugenol, ketamine and the ketamine/xylazine association allowed the deep anesthesia, but xilazine didn't allow it. Among the anesthetics tested, eugenol at 50mg/L demonstrated better anesthetic induction and recovery time for carp juveniles.

Nos procedimentos em piscicultura, os peixes são facilmente estressados nas diferentes etapas de criação. Assim, é importante o uso de anestésicos para prevenir injúrias físicas e promover a redução do metabolismo, resultando em melhor bem-estar aos peixes. O objetivo deste estudo foi analisar a indução anestésica em juvenis de carpa húngara (Cyprinus Carpio) expostos a diferentes doses de eugenol (20; 30; 40 e 50mg/L), ketamina (300; 600; 900 e 1200mg/L), xilazina (800; 1100; 2000mg/L) e da associação ketamina/xilazina (300/300; 600/600; 900/900 e 1200/1200mg/L). Cada dosagem foi testada em grupos de dez peixes escolhidos ao acaso (n=10), submetidos individualmente ao banho com anestésico em recipiente contendo 1 litro de água. Todos os procedimentos experimentais foram iguais nos ensaios. Após, foram analisados os tempos de anestesia (TA) e de recuperação (TR). Os anestésicos eugenol, ketamina e a associação ketamina/xilazina permitiram que os juvenis de carpa húngara atingissem a anestesia profunda. Entretanto, os dois últimos produtos testados causaram mortalidade em algumas doses. A xilazina não promoveu anestesia em nenhuma das doses testadas. Dentre os anestésicos testados, o eugenol na dose 50mg/L, demonstrou indução anestésica e tempo de recuperação satisfatório para juvenis de carpa.