Biomarkers and occurrences of heavy metals in sediment and the bioaccumulation of metals in crabs (Ucides cordatus) in impacted mangroves on the Amazon coast, Brazil.

In this study, we analyzed biomarkers and heavy metals in Ucides cordatus specimens caught in mangrove areas [A1 = reference (low impact area); A2, A3 and A4 = port complex (potentially impacted area)] of the Amazonian northern coast of Brazil. Sediments and crab muscles sampled from the potentially impacted mangroves showed high levels of metals. Additionally, catalase and glutathione S-transferase enzymes in the gills and hepatopancreas of the collected crabs were elevated in crabs from A2, A3 and A4 relative to those from A1. The histological characteristics of the gills and hepatopancreas differed in crabs collected from A2, A3 and A4 compared with those collected from A1. The crabs were larger and heavier in the rainy season in A1 and A3 and heavier in the dry season in A4. Heavy metal analyses of sediments, crab muscles and biological responses indicated that the animals are subjected to stress levels differentiated in the mangroves due to a gradient (A4>A3>A2>A1) of anthropogenic pressure in São Marcos Bay. In conclusion, the analyses of metals in the sediment and in the muscles of crabs, as well as the biochemical biomarker and histological analyses, suggest that crabs from mangroves in the port complex region are subject to pollutants that compromise their health.

Growth hormone on ovarian morphology of lambaris (Astyanax bimaculatus) after induced spawning / Hormônio de crescimento sobre a morfologia ovariana de lambaris (Astyanax bimaculatus) após desova induzida

ABSTRACT: Lambari, Astyanax bimaculatus, is an oviparous, multiple-spawning fish that is reproductively active throughout the year, which makes it promising for cultivation and research. This research histologically evaluates the ovaries of lambari that have undergone artificial spawning induced with pituitary extract (control group), and the effect of growth hormone at a dose of 2 mg/g body weight (treatment group) on the subsequent process of ovarian recovery. Ovaries of fish in both the control and treatments groups were collected at 120 hours after spawning and analyzed using optical microscopy to characterize the average quantities of: follicles in different stages of development, post-ovulatory follicles, follicular atresia and granulocytes. Quantity and morphology of early and advanced primary follicles did not differ between the treatment and control groups; an important and necessary factor for ovarian recovery for subsequent spawning. There was a greater amount of granulocytes in initial atresia in the group treated with growth hormone. These results demonstrated that the administration of growth hormone may potentiate the process of ovarian recovery after induced spawning.

RESUMO: O Lambari Astyanax bimaculatus é um peixe ovíparo de desova múltipla que é reprodutivamente ativo durante todo o ano, o que o torna promissor para cultivo e pesquisa. Este trabalho avalia histologicamente os ovários de lambaris submetidos à desova artificial, induzida pelo extrato hipofisário (grupo controle) e o efeito do hormônio de crescimento na concentração de 2 μg/g de massa corporal (grupo tratamento) no subsequente processo de recuperação ovariana. Os ovários dos peixes dos grupos controle e tratamento foram coletados às 120 horas após a desova e analisados em microscopia óptica para caracterizar as quantidades médias de: folículos em diferentes estágios de desenvolvimento, folículos pós-ovulatórios, atresia folicular e granulócitos. A quantidade e a morfologia dos folículos primários iniciais e avançados não diferiram entre os grupos tratamento e controle; um fator importante e necessário para a recuperação dos ovários para posterior desova. Houve maior quantidade de granulócitos na atresia inicial no grupo tratado com hormônio de crescimento. Esses resultados demonstram que a administração do hormônio do crescimento pode potencializar o processo de recuperação ovariana após a desova induzida.

Implications of body condition and seasonality on morphological and functional parameters of testes of Myotis nigricans (Chiroptera: Vespertilionidae).

The insectivorous bat Myotis nigricans is widely distributed throughout the Neotropics, including Brazil, and has a reproductive biology that is affected by climate and food availability. To evaluate the reproductive capacity of this species, morphofunctional parameters of the testes were correlated with environmental variables and the body condition of individuals captured. After bats had been killed, their testes were removed, fixed in Karnovsky's fluid for 24h and embedded in resin for evaluation by light microscopy. The mean annual tubulosomatic index (0.58%) and the percentage of seminiferous tubules in the testes (88.96%) were the highest ever recorded for the Order Chiroptera. The percentage of Leydig cells and volume of the cytoplasm of Leydig cells were higher in the rainy than dry season (80.62±3.19% and 573.57±166.95µm, respectively; mean±s.d.). Conversely, the percentage of nuclei of the Leydig cells in the dry season (26.17±3.70%; mean±s.d.) and the total number of Leydig cells (6.38±1.84×109; mean±s.d.) were higher in the dry season. The results of the present study could help in future conservation of these bats because they provide a better understanding of the bats' reproductive strategies and how the species can adapt to changes.

Immunohistochemical, morphological and histometrical analyses of follicular development in Astyanax bimaculatus (Teleostei: Characidae) exposed to an organochlorine insecticide.

Thiodan® is an organochlorine insecticide used in agriculture that can reach aquatic ecosystems where it can affect fish reproduction. This research aimed to evaluate follicular development and the expression of integrin ß1, collagen type IV and caspase 3 by morphological, histometrical and immunohistochemical analyses of Astyanax bimaculatus exposed to Thiodan®. Treatments included three sublethal concentrations of Thiodan® (1.15, 2.30, and 5.60µgL-1) for 96h and a control without the insecticide. Mature females with ovaries in advanced follicular development were chosen for study (average weight: 11.52 ± 2.0g; average total length: 9.12 ± 0.64cm). The follicles of A. bimaculatus exhibited normal morphology, while the diameters of secondary follicles showed an increase (P< 0.05) in all concentrations when compared to the control group; a characteristic of follicles undergoing the initial stages of intoxication. Immunohistochemical analysis revealed secondary follicles with greater expression of integrin ß1 and collagen type IV in cytoplasm of follicular cells than in the primary follicles in all treatments and in the control. Immunolocalization of caspase 3 was detected in follicular cells during secondary development in all tested concentrations of Thiodan® and the control. These analyses demonstrate positive immunolocalization throughout the course of follicular development, even in fish exposed to varying concentrations of Thiodan® for 96h, demonstrating that follicular cells retain their physiological integrity.

Anti-obesity effects of tea from Mangifera indica L. leaves of the Ubá variety in high-fat diet-induced obese rats.

Due to the high content of bioactive compounds, herbal teas are being investigated as adjuvant in chronic disease management. Studies have shown that mango leaf tea contain mangiferin, total phenolics and antioxidants, compounds with many functional properties. Therefore, this study aims to evaluate the anti-obesity effects of tea from Mangifera indica L. leaves, Ubá variety (TML), in obese rats fed a high-fat diet (HFD). For this, adult male Wistar rats were divided into three groups (n=8): the control group (fed AIN-93 diet), obese group (fed a HFD) and treated group (fed a HFD and supplemented with TML for 8 weeks). We analysed biometric measures and serum biochemical parameters of metabolic control, inflammation and oxidative stress biomarkers, histomorphometry of visceral adipose tissue and mRNA expression of peroxisome proliferator-activated receptor gamma co-activator 1 alpha (PPAR-γ), lipoprotein lipase (LPL) and fatty acid synthase (FAS). The consumption of TML (24.7±2.1mL/day) exerted antioxidant and anti-inflammatory effects, increasing total antioxidant capacity and interleukin-10 serum concentrations, reduced abdominal fat accumulation, upregulated PPAR-γ and LPL and downregulated FAS expression. Our data suggest that TML has therapeutic potential in treating obesity and related diseases through regulating the expression of transcriptional factors and enzymes associated with adipogenesis.

O inseticida Thiodan® sobre o desenvolvimento folicular de lambaris / Thiodan® insecticide on the follicular development of lambaris

O Thiodan(r) é um organoclorado a base de endosulfan que pode causar alterações morfológicas nos tecidos de peixes, dependendo da concentração e tempo de exposição. Este estudo teve como objetivo determinar a CL50-96h do Thiodan(r) (endosulfan 350g L-1) para fêmeas de lambaris Astyanax bimaculatus em período de reprodução e analisar a morfologia do desenvolvimento folicular em diferentes concentrações do agrotóxico. Foram feitos quatro experimentos: 1) sem aclimatação e sem alimentação; 2) sem aclimatação e alimentados; 3) com aclimatação de 10 dias e sem alimentação; 4) com aclimatação de 10 dias e alimentados. A CL50-96h determinada foi de 13,6µg L-1, com intervalo de confiança de 10,1 a 18,4µg L-1 (P<0,05). Em todos os experimentos, foram utilizadas três concentrações diferentes do Thiodan(r) inferiores à CL50-96h pré-determinada em laboratório de acordo com a NBR 15088 (ABNT, 2007). Os lambaris foram expostos ao Thiodan(r) por 96 horas em três concentrações subletais de 1,15; 2,3 e 5,6µg L-1 e um grupo controle, livre de agrotóxico. Morfologicamente, percebeu-se que a ação do Thiodan(r) nas concentrações subletais não alterou a morfologia do desenvolvimento folicular. Porém, o diâmetro folicular nos folículos secundários no experimento com aclimatação/com alimentação expostos ao Thiodan(r) foi maior em relação ao grupo controle (P<0,05). Esses dados sugerem que a reprodução pode ser afetada pelo produto químico e pode causar comprometimento no desenvolvimento folicular.

Thiodan(r) is an organochlorine based in endosulfan which can cause morphological changes in fish tissues exposed to it depending on the concentration and exposure time. This study aimed to determine the LC50-96h of Thiodan(r) (350g L-1 endosulfan) for lambaris females of Astyanax bimaculatus in a reproduction period and analyze the morphology of follicular development in different experiments. Four experiments were performed: without adaptation and no feeding, feeding without adaptation, with adaptation and without feeding, with adaptation and feeding. The LC50-96h determined was 13.6µg L-1, with a confidence interval from 10.1 to 18.4µg L-1 (P<0.05). The lambaris were exposed to Thiodan(r) for 96 hours in three sub-lethal concentrations of 1.15, 2.3 and 5.6µg L-1 and without pesticides control group. Morphologically, it was noted that the action of Thiodan(r) in sub-lethal concentrations did not alter the morphology of follicular development. However, the follicular diameter in secondary follicles in the experiment with adaptation and feeding exposed to Thiodan(r) was higher relative to the control group (P<0.05). These data suggest that reproduction may be affected by chemical and can cause impairment in the follicular development.

The addition of whole soy flour to cafeteria diet reduces metabolic risk markers in wistar rats.

BACKGROUND: Soybean is termed a functional food because it contains bioactive compounds. However, its effects are not well known under unbalanced diet conditions. This work is aimed at evaluating the effect of adding whole soy flour to a cafeteria diet on intestinal histomorphometry, metabolic risk and toxicity markers in rats. METHODS: In this study, 30 male adult Wistar rats were used, distributed among three groups (n = 10): AIN-93 M diet, cafeteria diet (CAF) and cafeteria diet with soy flour (CAFS), for 56 days. The following parameters were measured: food intake; weight gain; serum concentrations of triglycerides, total cholesterol, HDL-c, glycated hemoglobin (HbA1c), aspartate (AST) and alanine (ALT) aminotransferases and Thiobarbituric Acid Reactive Substances (TBARS); humidity and lipid fecal content; weight and fat of the liver. The villous height, the crypt depth and the thickness of the duodenal and ileal circular and longitudinal muscle layers of the animals were also measured. RESULTS: There was a significant reduction in the food intake in the CAF group. The CAFS showed lower serum concentrations of triglycerides and serum TBARS and a lower percentage of hepatic fat, with a corresponding increase in thickness of the intestinal muscle layers. In the CAF group, an increase in the HbA1c, ALT, lipid excretion, liver TBARS and crypt depth, was observed associated with lower HDL-c and villous height. The addition of soy did not promote any change in these parameters. CONCLUSIONS: The inclusion of whole soy flour in a high-fat diet may be helpful in reducing some markers of metabolic risk; however, more studies are required to clarify its effects on unbalanced diets.

Coadministration of sodium alginate pellets containing the fungi Duddingtonia flagrans and Monacrosporium thaumasium on cyathostomin infective larvae after passing through the gastrointestinal tract of horses.

The predatory nematophagous fungi have been used as an alternative control of gastrointestinal nematodes of domestic animals in natural and laboratory conditions. However, it is unclear if the association of some of these species could bring some kind of advantage, from a biological standpoint. In this context, this study consisted of two tests in vitro: in assay A, the assessment of the viability of the association of pellets in sodium alginate matrix containing the fungus Duddingtonia flagrans (AC001) and Monacrosporium thaumasium (NF34) and its predatory activity on infective larvae (L3) of cyathostomin after passing through the gastrointestinal tract of horses and assay B, assessment of the cyathostomin L3 reduction percentage in coprocultures. Twelve crossbred horses, females, with a mean weight of 356 kg and previously dewormed were divided in three groups with four animals each: group 1, each animal received 50 g of pellets containing mycelial mass of the fungus D. flagrans and 50 g of pellets of the fungus M. thaumasium, associated and in a single oral dose; group 2, 100 g of pellets containing D. flagrans and 100 g of pellets containing M. thaumasium, associated and in a single oral dose; group 3, control. Faecal samples were collected from animals in the treated and control groups at time intervals of 12, 24, 36, 48, 60 and 72 h after the administration of treatments and placed in Petri dishes containing 2% water-agar (assay A) and cups for coprocultures (assay B). Subsequently, 1000 cyathostomin L3 were added to each Petri dish (assay A) and 1000 cyathostomin eggs were added to each coproculture (assay B) of fungi-treated and control groups. At the end of 15 days, there was observed that the two associations of pellets containing the fungi tested showed predatory activity after passing through the gastrointestinal tract of horses (assay A). In assay B, all the intervals studied showed reduction rate in the number of L3 recovered from coprocultures exceeding 80%. However, no difference (p>0.01) was seen in recovery of not predated L3 between the fungi-treated groups in the time intervals studied. The results obtained showed that the associations of pellets (50 or 100 g of each fungal isolate) were viable after passage through the gastrointestinal tract in horses and could be used in natural conditions.

Increased production of biofilms by Escherichia coli in the presence of enrofloxacin.

The literature has demonstrated that subinhibitory concentrations of some antimicrobials are able to induce biofilm formation by certain bacterial species. Biofilms present in the mammary glands of cattle contribute to antimicrobial resistance, resulting in the appearance of persistent mastitis and consequent great losses to the dairy sector worldwide. The present study aimed to investigate the induction of biofilm formation by enrofloxacin in Escherichia coli isolates obtained from bovine mastitis. Twenty-seven isolates were reactivated in brain heart infusion (BHI) broth supplemented with different subinhibitory concentrations of enrofloxacin. Biofilm formation in microtiter plates was measured and confirmed by scanning electron microscopy (SEM). Isolates submitted to the concentration 0.0125 mg/mL of enrofloxacin showed greater biofilm formation compared to the control (p<0.001). Biofilm formation results obtained for the other concentrations did not differ from those obtained for the control (p>0.05). Using SEM it was possible to visualize the typical architecture of biofilms. These results represent the first report of inducing the production of biofilms in the presence of enrofloxacin, a quinolone antibiotic used to treat clinical mastitis.

Influence of the preservation period in silica-gel on the predatory activity of the isolates of Duddingtonia flagrans on infective larvae of cyathostomins (Nematoda: Cyathostominae).

The continued maintenance of nematophagous fungi predatory activity under laboratory conditions is one of the basic requirements for a successful biological control. The purpose of this study was to evaluate the influence of time on the preservation of the fungus Duddingtonia flagrans (AC001 and CG722) stored in silica-gel for 7 years and their subsequent predatory activity on cyathostomin L(3) larvae in 2% water-agar medium (2% WA). Samples of the isolates AC001 and CG722, originating from vials containing grains of silica-gel sterilized and stored for 7 years, were used. After obtaining fungal conidia, the predation test was conducted over 7 days on the surface of 9.0 cm Petri dishes filled with 2% WA. In the treated groups each Petri dish contained 500 cyathostomin L(3) and conidia of fungal isolates in 2% WA. In the control group (without fungi) the plates contained 500 L(3) in 2% WA. The experimental results showed that isolated AC001 and CG722 were efficient in preying on cyathostomin L(3) (p<0.01) compared to control (without fungus). However, no difference was observed (p>0.01) in the predatory activity of the fungal isolates tested. Comparing the groups, there was a significant reductions of cyathostomin L(3) (p<0.01) of 88.6% and 78.4% on average recovered from the groups treated with the isolates AC001 and CG722, respectively, after 7 days. The results of this test showed that the fungus D. flagrans (AC001 and CG722) stored in silica-gel for at least 7 years maintained its predatory activity on cyathostomin L(3).

The ovicidal activity of fungi Pochonia chlamydosporia and Paecilomyces lilacinus on Taenia saginata eggs in laboratory trial / Atividade ovicida dos fungos Pochonia chlamydosporia e Paecilomyces lilacinus sobre / ovos de Taenia saginata em condições laboratoriais

The ovicidal activity (type 3 effect) of fungi Pochonia chlamydosporia (VC1 and VC4 isolated) and Paecilomyces lilacinus (PL1) on Taenia saginata eggs was compared in an experimental trial. T. saginata eggs were fixed onto Petri dishes containing 2% water-agar (2% WA) and fungal isolates, and onto Petridishes without fungus as control. The ovicidal activity of these fungi was evaluated after being incubated for 5, 10 and 15 days. From the beginning of the interaction to the end of the trial, both P. chlamydosporia(VC1 and VC4) and P. lilacinus (PL1) fungi demonstrated ovicidal activity (p<0,05) when compared to the control. However, after 15 days of incubation the P. lilacinus fungus showed higher ovicidal activity (effect of type 3) when compared to P. chlamydosporia. This study showed that both fungi P. chlamydosporia (VC1and VC4) and P. lilacinus (PL1) are capable in killing eggs of T. saginata, therefore these fungi might be considered as potential candidates for biological control of this cestoda.

A atividade ovicida (efeito tipo 3) dos fungos Pochonia chlamydosporia (isolados VC1 e VC4) e Paecilomyceslilacinus (PL1) sobre os ovos de Taenia saginata foi investigada, em condições laboratoriais. Os ovos de T. saginata foram colocados em placas de Petri contendo o meio agar-água 2% (AA 2%) com os isolados fúngicos e, também, em placas de Petri sem fungos, como controle. A atividade ovicida desses fungos foi avaliada depois de 5, 10 e 15 dias de incubação. Ao final do experimento, os fungos P. chlamydosporia (VC1 e VC4) e P. lilacinus (PL1) demonstraram atividade ovicida (p<0,05) quando comparados com o controle. Contudo, aos 15 dias de incubação o fungo P. lilacinus demonstrou maior atividade ovicida (efeito do tipo 3) em relação ao P. chlamydosporia. Este estudo demonstrou que os fungos P. chlamydosporia (VC1 e VC4) e P. lilacinus (PL1) possuem a capacidade de destruir os ovos de T. saginata e, portanto, podem ser considerados como potenciais candidatos no controle biológico deste cestoda.

Parâmetros biométricos maternos e fetais e suas relaçöes com diâmetros obstétricos durante a gestaçäo em cabras mestiças / Biometric maternal and fetal parameters and its relations with obstetrics diameters during the gestation in crossbred goats

Foram obtidos dados biométricos de cabras mestiças aos 50, 100 e 140 dias de gestaçäo e dos respectivos fetos para estudar-se a relaçäo entre peso, comprimento céfalo-caudal (CCR) e profundidade torácica (PT) maternos e fetais, além de medidas externas e internas dos diâmetros obstétricos conjugado (DC) e transverso (DT) da pelve materna. Observou-se que as alteraçöes das medidas de CCR, PT, DT externo e DCs interno e externo das fêmeas, acompanharam significamente a variaçäo do peso corporal da mesma e refletiram sobre o peso, CCR e PT de fetos com 50 e 140 dias de gestaçäo. A PT da fêmea relacionou-se significamente apenas com o peso e CCR das mesmas. Verificou-se que as alteraçöes observadas nas medidas externas da pelve refletiram sobre as internas, indicando que mensuraçöes externas correspondentes às regiöes da pelve auxiliam na determinaçäo aproximada da área interna da mesma. A variaçäo do peso da fêmea exerceu influência sobre todas as medidas corporais do feto, auxiliando na determinaçäo do tamanho aproximado do mesmo. O sexo do feto näo influenciou suas medidas corporais e tanto os machos quanto as fêmeas apresentaram tamanhos semelhantes durante a gestaçäo, indicando que o sexo näo se relaciona com a dificuldade no parto