Acute sublethal exposure to ethiprole impairs physiological and oxidative status in the Neotropical fish Astyanax altiparanae.

Ethiprole, a phenylpyrazole insecticide, has been increasingly used in the Neotropical region to control stink bug pests in soybean and maize fields. However, such abrupt increases in use may have unintended effects on non-target organisms, including those inhabiting freshwater ecosystems. Here, we evaluated the effects of acute (96 h) sublethal exposure to ethiprole (up to 180 µg/L, which is equivalent to 0.013% of the recommended field dose) on biomarkers of stress in the gills, liver, and muscle of the Neotropical fish Astyanax altiparanae. We further recorded potential ethiprole-induced effects on the structural histology of A. altiparanae gills and liver. Our results showed that ethiprole exposure increased glucose and cortisol levels in a concentration-dependent manner. Ethiprole-exposed fish also exhibited higher levels of malondialdehyde and greater activity of antioxidant enzymes, such as glutathione-S-transferase and catalase, in both gills and liver. Furthermore, ethiprole exposure led to increased catalase activity and carbonylated protein levels in muscle. Morphometric and pathological analyses of the gills revealed that increasing ethiprole concentration resulted in hyperemia and loss of integrity of the secondary lamellae. Similarly, histopathological analysis of the liver demonstrated higher prevalence of necrosis and inflammatory infiltrates with increasing ethiprole concentration. Altogether, our findings demonstrated that sublethal exposure to ethiprole can trigger a stress response in non-target fish species, which may lead to potential ecological and economic imbalances in Neotropical freshwater systems.

Sperm characteristics, peroxidation lipid and antioxidant enzyme activity changes in milt of Brycon orbignyanus cryopreserved with melatonin in different freezing curves.

The objective of this study was to evaluate extender supplemented with melatonin and freezing curves on the antioxidant enzyme activity, peroxidation lipid and sperm characteristics of cryopreserved Brycon orbignyanus milt. Males (n = 16) and females (n = 5) were hormonally induced with two doses (0.5 mg and 5.0 mg kg-1) of carp pituitary extract, and their gametes were collected by light abdominal massage. The fresh milt was diluted at a ratio of 1:4 (milt:extender) in the following solutions: (Control) 10% methyl glycol (MG) + 5% Beltsville thawing solution; (M1) Control + 1 mM melatonin; and (M2) Control + 2 mM melatonin. The freezing curves were C1 (automated freezer) and C2 (dry shipper for 24 h). After each curve was recorded, the straws were transferred to a liquid nitrogen container until the analyses were performed. The samples were thawed in a water bath (60 °C, 8 s) and evaluated using the Sperm Class Analyzer software for the parameters total motility, progressive motility, curvilinear velocity, straight line velocity, mean displacement velocity, straightness, and linearity. The following were also measured: motility time, vitality, morphology, oxidative stress (lipid peroxidation, activity of catalase and superoxide dismutase, quantification of nitric oxide), and fertilization and hatching rates. The data were analyzed within R by one-way analysis of variance and Tukey's test for comparison of means (p < 0.05). A significant difference (p < 0.05) was observed between the solutions in vitality, morphology, motility, and fertilization rate, the solutions with melatonin having the best values. Total motility, progressive motility, and motility time were significantly different. Among oxidative stress markers, only lipid peroxidation and superoxide dismutase activity showed an effect of the curve × solution interaction (p < 0.05), the solutions with melatonin yielding the lowest values. The fertilization and hatching rates were also higher under the melatonin treatments, regardless of the curve. Melatonin 2 mM and slow curve are indicated for the cryopreservation of fish species sperm as it led to the slowest detrimental spermatozoa effects and better fertilization and hatching rates.

Tebuconazole-induced toxicity and the protective effect of Ficus carica extract in Neotropical fruit-eating bats.

Tebuconazole (TEB) is a triazole fungicide widely used in agriculture known to cause metabolic and endocrine disorders in mammals. Several plant extracts have shown to be beneficial against pesticide effects due to their hepatoprotective, antioxidant and anti-inflammatory properties. As fruit bats play a critical role in rainforest regeneration and are constantly exposed to pesticides, we aimed at evaluating TEB-induced toxicity and the possible protective effect of the Ficus carica plant extract in Neotropical fruit-eating bats (Artibeus lituratus). Bats were captured and assigned to 4 experimental groups, offered: 1) CTL (n = 6): papaya; 2) DMSO (n = 6): papaya treated with 1.25% dimethyl sulfoxide (DMSO); 3) TEB (n = 6): papaya treated with tebuconazole (commercial formulation) 0.1%; and 4) TEBFC (n = 6): papaya treated with tebuconazole 0.1% and Ficus carica extract (20%) in DMSO (1.25%). After seven days of exposure, TEB bats showed increased lipid peroxidation, increased superoxide dismutase (SOD) and catalase (CAT) activities, vascular congestion and inflammatory infiltrate in the liver, and increased serum transaminase enzyme activities. We found the same alterations in oxidative stress parameters in the breast muscles of TEB-exposed bats. In the testes, all oxidative stress markers were increased in TEB bats and corroborate findings of histopathological and increased serum testosterone levels observed following TEB exposure. The co-administration of the fungicide with the F. carica plant extract attenuated most oxidative stress markers in exposed bats' liver and testes and decreased liver damage, but failed to revert the steroid imbalance caused by the fungicide exposure.

Effects of subchronic exposure to environmentally relevant concentrations of the herbicide atrazine in the Neotropical fish Astyanax altiparanae.

Atrazine (ATZ) is among the most widely used herbicides in the world, and yet it has a potential to contaminate aquatic environments due to pesticide leaching from agricultural areas. In the Neotropical region, studies about the effects of this herbicide in native aquatic wildlife is scarce.Our study aimed at investigating the effects of a 30-day exposure to a commercial atrazine formulation on oxidative stress parameters, histopathology in testis and liver, and hormone levels in males and female of yellow-tailed tetra fish (Astyanax altiparanae). Adults were exposed to low but environmentally relevant concentrations of atrazine as follows: 0 (CTL-control), 0.5 (ATZ0.5), 1 (ATZ1), 2 (ATZ2) and 10 (ATZ10) µg/L. Our results showed decreased GST activity in gills in all groups of exposed animals and increased CAT activity in gills from the ATZ10 group. In the liver, there was an increase in lipid peroxidation in fish from ATZ1 and ATZ2 groups. Histological analysis of the liver showed increased percentage of sinusoid capillaries in ATZ2 fish, increased vascular congestion in ATZ1 and increased leukocyte infiltration in the ATZ10 group. Hepatocyte diameter analysis revealed a decrease in cell size in all groups exposed to ATZ, and a decrease in hepatocyte nucleus diameter in ATZ1, ATZ2 and ATZ10 groups. Endocrine parameters did not show significant changes following ATZ exposure, although an increase of triiodothyronine/thyroxine (T3/T4) ratio was observed in ATZ2 fish. Our results provide evidence that even low, environmentally relevant concentrations of ATZ produced oxidative damage and histological alterations in adult yellow-tailed tetra.

"Fearing the enemy": Growth and stress biomarker responses of sexually reversed Oreochromis niloticus in the presence of aquatic predatory insects.

Fishes can change their physiological responses when threatened by the presence of predators. Such physiological plasticity, however, usually implies costs that may impede organismal development and reproduction and reduce the ability to cope with other biotic and abiotic stresses. Here, we evaluated the growth and stress biomarker responses in sexually reversed Nile tilapia, Oreochromis niloticus, fingerlings indirectly threatened by the presence of the aquatic insect predator Belostoma anurum (Hemiptera: Belostomatidae). We also evaluated whether the presence of B. anurum would affect growth in fingerlings that received food containing the masculinizing hormone 17 α-methyltestosterone. The antioxidant responses were evaluated by measuring the activity of enzymes (e.g., superoxide dismutase, catalase, and glutathione-S-transferase). Oxidative stress biomarkers (e.g., malondialdehyde and nitric oxide) and blood glucose and lactate responses were also evaluated. Our results revealed that predator exposure did not affect growth in O. niloticus fingerlings reared in the presence or absence of the masculinizing hormone. However, sexually reversed tilapia fingerlings significantly increased not only the glucose and lactate blood levels, but also exhibited increased activities of superoxide dismutase and glutathione-S-transferases enzymes when threatened by the presence of B. anurum nymphs. Collectively, our findings indicate that despite not exhibiting reduced growth performance, sexually reversed tilapia fingerlings were physiologically stressed by the presence of the predator, which may reduce their ability to face environmental and abiotic stresses.

Impacts of subchronic exposure to a commercial 2,4-D herbicide on developmental stages of multiple freshwater fish species.

Invasive, nuisance aquatic species such as Eurasian watermilfoil (Myriophyllum spicatum) are rapidly spreading across the United States. One common active ingredient used to control this invasive species is 2,4-Dichlorophenoxyacetic acid (2,4-D). Application of 2,4-D to aquatic environments typically occurs while many freshwater fish are spawning and due to 2,4-D stability in aquatic environments, many non-target species experience prolonged exposure throughout embryogenesis and larval development. The impacts of 2,4-D exposure on phylogenetically distant fish species is poorly understood. Herein, we investigated the impacts of the 2,4-D commercial herbicide DMA4®IVM on nine freshwater fish species from six different families (four orders) at different points during ontogeny. Each species was exposed to ecologically relevant concentrations of a commercial 2,4-D herbicide (0.05, 0.50, and 2.00 ppm or mg/L 2,4-D a. e.), and effects on morphology, survival, and growth were evaluated. Our results demonstrate that exposure of embryonic and larval fish to ecologically relevant concentrations of a commercial 2,4-D herbicide reduced survival in early developmental stages of six freshwater species that spanned five phylogenetic families and three phylogenetic orders; however, sensitivity to 2,4-D exposure did not show correlation with phylogenetic proximity. Altogether, our results indicate that the use of 2,4-D herbicides in aquatic ecosystems at current recommended concentrations (≤2 ppm whole-lake treatment) could present risk to multiple freshwater fish species survival during early development.

Is bone loss a physiological cost of reproduction in the Great fruit-eating bat Artibeus lituratus?

During mammalian pregnancy and lactation, the maternal demand for calcium is increased to satisfy fetus and newborn skeletal growth. In addition to the dietary intake, females use the calcium contained in their bones to supply this increased demand, leading to a decrease in maternal bone mineral content. In reproductive insectivorous female bats, bone loss has been described as a physiological cost of reproduction, due to the reported increased risk of bone fracture. This physiological cost may be the mechanism underlying the conflict between increasing litter size and maintaining wing skeletal integrity, which would help to explain the small litter size of most bat species. If bone loss is a linking cost between reproduction and survival in bats, and most bat species have small litter sizes, one would expect to find a loss of bone and an increasing probability of bone fracture during pregnancy and lactation in other non-insectivorous bats. In this study, we tested for the existence of this cost in the Great-fruit eating bat, Artibeus lituratus. We analyzed trabecular structure, bone strength and bone mineral content for the humerus bone, hypothesizing that bone loss during reproduction in females would increase the risk of fracture. Our results showed a decrease of 22-31% in bone trabecular area in lactating females, rapidly compensated following weaning. Bone strength did not differ among reproductive and non-reproductive groups and seems to be more influenced by bone organic components rather than mineral contents. Since we observed bone loss during reproduction yet the humerus strength seems to be unaffected, we suggest that bone loss may not represent a physiological cost during reproduction for this frugivorous bat.

Haematological alterations in non-human hosts infected with Trypanosoma cruzi: a systematic review.

American trypanosomiasis is a neglected tropical disease whose spectrum has not been quite understood, including the impact of Trypanosoma cruzi infection on the haematological parameters of different vertebrate hosts. Thus, this study was designed to compare the pattern of haematological changes induced by T. cruzi infection in order to identify possible species-specific differences among taxons. We also aimed at evaluating the use of this parameter as a tool for diagnosis during the acute phase, when symptoms are usually masked. For this purpose, we performed a systematic search on PubMed and Scopus databases to retrieve original studies published until August 2016. Thirty-one studies were selected using Prisma strategy, which were then submitted to data extraction and methodological bias analysis. Half of the studies showed that the number of erythrogram decreased in infected animals, indicating anaemia. In 68.2% of the studies, the total amount of leukogram values increased, suggesting infection. The main methodological limitations were insufficient information for T. cruzi strains identification, inoculation routes and parasitological characterization. Most of the mammalian species analysed showed the same pattern of haematological changes following T. cruzi infection, indicating that haematological parameters might direct the diagnosis of Chagas disease in the initial phase.

Dietary Exposure to Tebuconazole Affects Testicular and Epididymal Histomorphometry in Frugivorous Bats.

This study evaluated the effects of a commercially recommended concentration (1 mL/L) of a fungicide tebuconazole (TBZ) on testicular and epididymal histomorphometry of Artibeus lituratus, following 7 and 30-day oral exposure. TBZ30 bats showed a reduction in the percentage of tubules and seminiferous epithelium, as well as a decrease in tubule and epithelium somatic indexes, and tubular diameter. Inversely, these animals showed increased percentage of intertubular compartment, Leydig cells and blood vessels. The volume of Leydig cells and their number per gram of testis also increased in TBZ30 bats. Alterations in epididymal morphometry were observed in all regions of the organ, with increase of ductal diameter in both exposure times. These results indicate that exposure to low concentration of TBZ resulted in testicular and epididymal morphometric changes in fruit bats, mainly at 30-day exposure, suggesting that functional alterations might be occurring in these organs and impacting reproductive capacity.

Effects of low, subchronic exposure of 2,4-Dichlorophenoxyacetic acid (2,4-D) and commercial 2,4-D formulations on early life stages of fathead minnows (Pimephales promelas).

Aquatic herbicides are commonly used to control a wide variety of algae and plants, but they also have the potential to contaminate and affect nontarget organisms. However, the impacts of low-level 2,4-dichlorophenoxyacetic acid (2,4-D) herbicide exposure on larval fish are not well understood. We conducted a series of experiments to determine the effects of low concentrations (0.05, 0.50, and 2.00 ppm) of 2 commercial 2,4-D amine salt herbicide formulations (Weedestroy® AM40 [WAM40] and DMA® 4 IVM [DMA4]) and pure 2,4-D on the development and survival of fathead minnows (Pimephales promelas) at various life cycle stages. Larval survival (30 d post hatch [dph]) was decreased following exposure of eggs and larvae to pure 2,4-D (0.50 ppm; p ≤ 0.001), as well as to WAM40 (0.50 and 2.00 ppm; p ≤ 0.001, p ≤ 0.001) and DMA4 (0.50 and 2.00 ppm; p ≤ 0.001, p ≤ 0.001). The results also narrowed the critical window of exposure for effects on survival to the period between fertilization and 14 dph. Development was not negatively altered by any of the compounds tested, although the commercial formulations increased larval total length and mass at 2.00 ppm. Altogether, the results indicate that the use of 2,4-D herbicides for weed control in aquatic ecosystems at current recommended concentrations (<2 ppm whole lake; <4 ppm spot treatment) could present risks to fathead minnow larval survival. Environ Toxicol Chem 2018;37:2550-2559. © 2018 SETAC.

Changes in bone turnover and calcium homeostasis during pregnancy and lactation in mammals: a meta-analysis.

Large amounts of calcium are required during pregnancy and lactation to support fetal and neonatal bone growth and calcification. An inadequate supply of calcium during these stages can lead to unsuccessful reproduction or impaired offspring fitness. During reproduction, female mammals undergo numerous physiological changes, including adaptations to allow an adequate supply of calcium. The lack of quantitative studies analysing these physiological changes from a comparative perspective limits our ability to explain and understand these adaptations. Herein, we present our meta-analysis of studies reporting changes in bone turnover and calcium homeostasis during pregnancy and lactation in 14 species of mammals. Our meta-analysis of 60 studies showed that all species have a similar pattern of physiological changes during pregnancy and lactation, which include: (1) decreased serum calcium concentrations; (2) bone tissue loss; (3) decreased serum calcitonin and parathyroid hormone concentrations; and (4) increased serum calcitriol concentration, regardless of changes in parathyroid hormone concentrations. In addition, we found a negative relationship between: (1) serum calcium concentrations and the number of teats; and (2) serum parathyroid hormone concentrations and litter mass.

The expression patterns of aquaporin 9, vacuolar H+-ATPase, and cytokeratin 5 in the epididymis of the common vampire bat.

Desmodus rotundus is a vampire bat species that inhabits Latin America. Some basic aspects of this species' biology are still unknown, as the histophysiological characteristics of the male reproductive tract. Our study has focused on its epididymis, which is an important organ for performing a variety of functions, especially the sperm maturation and storage. The aim of this study was to identify principal, narrow, clear, and basal cells using cell-specific markers such as aquaporin 9 (AQP9), vacuolar H+-ATPase (V-ATPase), and cytokeratin 5 (KRT5). Principal cells were labeled by AQP9 from initial segment to cauda region in their stereocilia. They were shown with a columnar shape, whereas V-ATPase-rich cells were identified with a goblet-shaped body along the entire epididymis, including the initial segment, which were named as clear cells. Pencil-shaped V-ATPase-rich cells (narrow cells) were not detected in the initial segment of the bat epididymis, unlike in the rodent. Basal cells were labeled by KRT5 and were located at the basal portion of the epithelium forming a dense network. However, no basal cells with a luminal-reaching body extension were observed in the bat epididymis. In summary, epithelial cells were identified by their specific markers in the vampire bat epididymis. Principal and basal cells were labeled by AQP9 and KRT5, respectively. Narrow cells were not observed in the vampire bat epididymis, whereas clear cells were identified by V-ATPase labeling along the entire duct in a goblet-shaped body. In addition, no luminal-reaching basal cells were observed in the vampire bat epididymis.

Warmer temperature modifies effects of polybrominated diphenyl ethers on hormone profiles in leopard frog tadpoles (Lithobates pipiens).

Amphibian populations have been declining, and climate change and exposure to environmental contaminants are thought to be involved. Higher water temperature accelerates larval development; however, its combined effects with contaminants and their influence on hormones during metamorphosis are poorly understood. The authors investigated changes in whole-body triiodothyronine (T3) and corticosterone concentrations in developing leopard frogs reared at 23 °C and 28 °C on diets with 0 ng g-1 , 6 ng g-1 , and 37 ng g-1 of a technical mixture of polybrominated diphenyl ethers (PBDE; DE-71) from 10 d to 44 d (premetamorphosis to late climax; Gosner Stages 28 to 46). Unlike controls, PBDE-exposed tadpoles (6 ng g-1 ) reared at 23 °C failed to show any increase in T3 concentrations throughout metamorphosis, and exposed tadpoles reared at 28 °C showed a lower peak at climax compared to controls. Corticosterone levels progressively increased throughout metamorphosis, but the levels were higher in PBDE-exposed tadpoles compared to controls at both temperatures. At the warmer temperature, corticosterone increase occurred earlier (at early climax) in controls and exposed tadpoles compared to tadpoles reared at the cooler temperature (late climax), coinciding with the faster development observed at 28 °C. Tadpoles reared at 28 °C were longer and developed faster than tadpoles reared at 23 °C. At both temperatures, PBDE exposure decreased T3 and increased corticosterone concentrations, which can potentially impair developing tadpoles. Environ Toxicol Chem 2017;36:120-127. © 2016 SETAC.

5α-Dihydrotestosterone enhances wound healing in diabetic rats.

UNLABELLED: Wound healing involves a complex interaction between the cells, extracellular matrix and oxidative response. AIMS: Analyze the effects of 5α-Dihydrotestosterone (5α-DTH) ointment in cutaneous wound healing by secondary intention in diabetic Wistar rats. MAIN METHODS: Rats (302.23±26.23g, n=48) were maintained in cages with food and water ad libitum in accordance with the Guiding Principles in the Use of Animal Ethics Committee. Diabetes was induced by intraperitoneal injection of streptozotocin (60mg/kg). Three skin wounds (12mm diameter) were created on the animals' back, which were randomized into 6 groups according to the application received: VT group: Vehicle (lanolin), SA group: 0.9% saline solution, NC group: Non-diabetic, CP group: positive control (silver sulfadiazine 0001%), T1 group: Testosterone (10%), T2 group: Testosterone (20%) emulsified in lanolin. The applications were made daily within 21days, and tissues from different wounds were removed every 7days. KEY FINDINGS: Both groups treated with testosterone (T1 and T2) showed a significantly higher proportion of type I and type III collagen fibers. Superoxide dismutase levels were significantly higher on days 7 and 14 in testosterone treated groups. Protein carbonyls and MDA were lower in both groups. SIGNIFICANCE: We conclude that groups treated with 5α-DTH showed a better healing pattern with complete wound closure, and proved to have a positive effect on the morphology of the scar tissue as well as an antioxidant stimulating effect during secondhand intention skin wounds repair in diabetic rats.

Evaluation of the cell population of the seminiferous epithelium and spermatic indexes of the bat Sturnira lilium (Chiroptera: Phyllostomidae).

Due to the scarcity of information about patterns of spermatogenesis in bats, this study aimed to provide information on the testicular activity of the bat Sturnira lilium along the annual seasons. Thus, a series of morphometrical and stereological analyses were made using the testes of adult S. lilium in order to achieve a better understanding of the sperm production dynamics. Light and transmission electron microscopy analyses were performed in testicular fragments of animals captured during dry and rainy seasons. The testes followed the pattern of organization described for other mammals, and there were no morphological differences between organs collected either in dry or in rainy seasons. Each tubular cross-section in stage 1 was made of 0.5 type-A spermatogonia, 4.4 primary spermatocytes in preleptotene/leptotene, 3.7 in zygotene, 11.9 in pachytene, 35.6 round spermatids and 8.5 Sertoli cells. The mitotic and meiotic indexes were 15.4 and 2.9 cells, respectively, while the spermatogenesis yield was 68.7 cells. The testicular sperm reserves was 37.61×10(6) cells, and daily sperm production per gram of testis averaged 209.68×10(6) cells, both highest averages occurring in the rainy season. S. lilium male bats have a continuous reproductive pattern, high spermatogenesis yield and low support capacity by the Sertoli cells.

Histomorphometric characterization of the intertubular compartment in the testes of the bat Sturnira lilium.

Due to the scarcity of information about the reproduction of bats, it is necessary to perform studies on different species to identify the occurrence of annual variations in their reproductive process. Therefore, the aims of this study were to describe and quantify the intertubular components within the testes of the bat Sturnira lilium and to verify whether seasonality takes place in spermatogenesis of this species. The animals were collected in different seasons and time of collection was grouped into dry and rainy seasons. Testicular fragments were routinely processed for light and transmission electron microscopies and blood samples from each animal were collected for quantification of plasma testosterone. Overall, the Leydig cells (LC) were the main intertubular component (83.2%), with abundant lipid droplets in their cytoplasm. Ultrastructural analysis indicated collagen fibers in the connective tissue and lymphatic spaces, with thin walls, surrounding the seminiferous tubules. The proportion and volume of each intertubular component did not vary significantly between seasons. On average, testosterone concentrations did not vary between rainy and dry seasons (21ng/mL). The total number of LC in the testis (50.0×10(5)) and per gram of testis (11×10(7)) did not vary nor did the Leydigosomatic Index (0.03%). Therefore, it is concluded that S. lilium had significant investment in intertubular tissues, especially in LC. Most of the variables that were assessed did not vary with season of the year, which leads to the assumption that S. lilium has a continuous reproductive cycle in southeastern Brazil.

Histomorphometric evaluation of the Molossus molossus (Chiroptera, Molossidae) testis: the tubular compartment and indices of sperm production.

Insectivorous bats play a very important role in the regulation of tropical ecosystems, but information about their reproductive cycle is lacking. Thus, male Molossus molossus were captured over the four seasons, and morphometric analyses of their testes were conducted to infer on the gonadal dynamics and the reproductive capacity of the species. Testes were immersed in Karnovsky fixative, and fragments were embedded in methacrylate and paraplast for morphometric and TUNEL assay respectively. The least gonadosomatic index (0.3%), tubulesomatic index (0.2%) and tubular diameter (133.2µm) occurred in summer. An adult M. molossus showed a total average of 48.9m of seminiferous tubules per gram of testis. Primary spermatocytes were observed in the zygotene at Stage 1 of the seminiferous epithelium cycle. The greatest meiotic index was obtained in winter (3.8 cells), and the general yield of spermatogenesis was higher in winter (64.5 cells) than in summer (19.1 cells). There was no difference in the apoptotic cells count among seasons. The Sertoli cell index was less in summer (5.9) than in fall (11.6), while the number of Sertoli cells per gram of testis did not vary significantly among the seasons (28.0×10(7)). The spermatic reserve per gram of testis was greater in the fall (63.9×10(7)) and winter (69.8×10(7)) than summer (37.1×10(7)). We conclude that M. molossus males show a continuous reproductive cycle, featuring greater spermatogenic activity during the fall and winter, a tubular length above the average of other mammals and a less support capacity of the Sertoli cells.

Stages and duration of the seminiferous epithelium cycle in the bat Sturnira lilium.

Knowledge of the stages that compose the seminiferous epithelium cycle (SEC) and determination of the duration of spermatogenic processes are fundamental for the accurate quantification of the dynamics of spermatogenesis. The aim of this study was to characterize the stages that compose the SEC of the bat Sturnira lilium, including evaluation of the average frequency of each of these stages throughout the year and calculation of the duration of the spermatogenic process. An ultrastructural characterization of the formation of the acrosomal cap was also performed. Testicular fragments were processed for morphological and immunohistochemical analysis as well as ultrastructural analysis using transmission electron microscopy. According to the tubular morphology method, the SEC in S. lilium is divided into eight stages, following the pattern found in other mammals. Primary spermatocytes were found at zygotene in stage 1 of the cycle. There was no variation in frequency of each of the stages over the seasons, with stage 1 being the most frequent, and stage 7 the least frequent. The duration of one seminiferous epithelium cycle was 3.45 days, and approximately 15.52 days were required for the development of sperm from spermatogonia. Ultrastructural characterization allowed the formation of the acrosomal cap in round spermatids to be monitored. In conclusion, the stages that compose the SEC in S. lilium are generally similar to those described for other mammals, but the duration of the spermatogenic process is shorter than previously recorded for mammals. The presence of primary spermatocytes at zygotene in stage 1 of the cycle is probably due to the longer duration of this stage.

Reduced insulin secretion and glucose intolerance are involved in the fasting susceptibility of common vampire bats.

Susceptibility during fasting has been reported for the common vampire bat (Desmodus rotundus), to the point of untimely deaths after only 2-3 nights of fasting. To investigate the underlying physiology of this critical metabolic condition, we analyzed serum insulin levels, pancreatic islets morphometry and immunocytochemistry (ICC), static insulin secretion in pancreas fragments, and insulin signaling mechanism in male vampire bats. A glucose tolerance test (ipGTT) was also performed. Serum insulin was found to be lower in fed vampires compared to other mammals, and was significantly reduced after 24h fasting. Morphometrical analyses revealed small irregular pancreatic islets with reduced percentage of ß-cell mass compared to other bats. Static insulin secretion analysis showed that glucose-stimulated insulin secretion was impaired, as insulin levels did not reach significance under high glucose concentrations, whereas the response to the amino acid leucin was preserved. Results from ipGTT showed a failure on glucose clearance, indicating glucose intolerance due to diminished pancreatic insulin secretion and/or decreased ß-cell response to glucose. In conclusion, data presented here indicate lower insulinemia and impaired insulin secretion in D. rotundus, which is consistent with the limited ability to store body energy reserves, previously reported in these animals. Whether these metabolic and hormonal features are associated with their blood diet remains to be determined. The peculiar food sharing through blood regurgitation, reported to this species, might be an adaptive mechanism overcoming this metabolic susceptibility.