Eugenol influences the expression of messenger RNAs for superoxide dismutase and glutathione peroxidase 1 in bovine secondary follicles cultured in vitro.

This study aimed to investigate the effects of eugenol on growth, viability, antrum formation and mRNA expression of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase 1 (GPX1) and peroxiredoxin 6 (PRDX6) in bovine secondary follicles cultured in vitro. To this end, bovine ovaries were collected from a local slaughterhouse and in the laboratory the follicles were isolated from the ovarian cortex. The follicles were then cultured in TCM-199+ alone or supplemented with different concentrations of eugenol (0.5, 5.0 and 50.0 µM). Follicular diameters and antrum formation were evaluated on days 0, 6, 12 and 18. Viability analysis was performed using calcein and ethidium homodimer. Real-time PCR was used to quantify mRNA levels for SOD, CAT, GPX1 and PRDX6 in cultured follicles. Follicular diameters and mRNA levels in follicles cultured in vitro were compared using analysis of variance and Kruskal-Wallis tests, while follicular survival and antrum formation were compared using the chi-squared test (P < 0.05). The results showed that secondary follicles cultured with eugenol maintained similar morphology and viability to follicles cultured in the control group. A progressive increase in follicular diameter was observed between days 0 and 12 for all treatments, except for follicles cultured with 50 µM eugenol. Eugenol (5.0 and 50.0 µM) increased mRNA levels for GPX1 in cultured follicles, but 0.5 µM eugenol reduced mRNA levels for SOD. The addition of eugenol did not influence mRNA expression for CAT and PRDX6. In conclusion, eugenol supplementation reduces mRNA levels for SOD and increases mRNA levels of GPX1 in bovine secondary follicles cultured in vitro.

Effects of dexamethasone on growth, viability and ultrastructure of bovine secondary follicles cultured in vitro.

This study aimed to evaluate the effects of dexamethasone on development, viability, antrum formation and ultrastructural integrity of bovine secondary follicles cultured in vitro for 18 days. Bovine ovaries were obtained from slaughterhouses and secondary follicles of ~150-200 µm diameter were isolated and cultured in the laboratory in TCM-199+ alone or supplemented with different concentrations of dexamethasone (1, 10, 100 and 1000 ng/ml). Follicle viability was evaluated after the culture period, using calcein-AM (viable) and ethidium homodimer (nonviable). Follicle diameters and antrum formation were evaluated at days 0, 6, 12 and 18. Before or after in vitro culture, follicles were fixed for histological and ultrastructural analysis. Follicle diameters were evaluated using analysis of variance and Kruskal-Wallis test, while chi-squared test was used to evaluate the percentage of viable follicles and antrum formation (P < 0.05). Follicles cultured for 6 days with all treatments increased their diameters significantly, but there was no significant difference between treatments at the end of the culture period. In vitro cultured follicles showed antral cavity formation at the end of the culture period, but no influence of dexamethasone was seen. Ultrastructural analysis showed that follicles cultured with dexamethasone (1, 10, 100 and 1000 ng/ml) had well preserved granulosa cells. However, oocytes from follicles cultured with 10, 100 or 1000 ng/ml dexamethasone showed signs of degeneration. It can be concluded that follicles cultured in vitro in the presence of dexamethasone demonstrated continuous in vitro growth, but oocytes from follicles cultured with 10, 100 or 1000 ng/ml dexamethasone had poor ultrastructure.

Supplementation of culture medium with knockout serum replacement improves the survival of bovine secondary follicles when compared with other protein sources during in vitro culture.

The present study evaluated the effect of knockout serum replacement (KSR), fetal bovine serum (FBS) and bovine serum albumin (BSA) on the viability and growth of bovine secondary follicles cultured in vitro for 12 days. To this end, secondary follicles were isolated (185-202 µm) and cultured in vitro in TCM-199+ medium supplemented with KSR (5% and 10%), FBS (5% and 10%) or BSA (3 mg/ml) at 38.5°C with 5% CO2 in air. Follicular diameters were evaluated on days 0, 4, 8 and 12. After 12 days of culture, follicular survival analysis was performing by using calcein-AM and ethidium homodimer. Before and after culture, follicles were fixed in paraformaldehyde for histological evaluation. Follicular diameter at different days of culture were compared using the Kruskal-Wallis test, while the percentages of viable follicles were analyzed by chi-squared test (P < 0.05). Results showed that follicles cultured in the presence of KSR at both concentrations presented higher follicular survival rates than those cultured in control medium alone or supplemented with FBS or BSA. Conversely, the presence of KSR, BSA or FBS did not increase follicular diameter after 12 days of culture. Histology analysis showed that, among the tested treatments, follicles cultured in the presence of KSR had preserved rounded oocytes, juxtaposed granulosa cells and intact basal membrane. In conclusion, supplementation of culture medium with KSR increases the follicular survival of bovine secondary follicles cultured in vitro.

Delinted cottonseed in diets for finishing sheep.

The use of agricultural by-products might provide an important alternative to improve animal agriculture and to reduce feeding costs. The effects of inclusion in the diet of four levels of delinted cottonseed (0, 80, 160, and 240 g/kg) on intake, digestibility, nitrogen balance, blood parameters, and the ingestive behaviour of confined sheep were studied. Twenty-four mixed-breed rams with an average body weight of 27.6 ± 2.9 kg were used in a completely randomised design with four treatments. The addition of delinted cottonseed (DCS) did not alter (P > 0.05) the intake of dry matter, crude protein, organic matter, neutral detergent fibre, or ether extract in the two forms calculated (g/day and % body weight). The use of different levels of DCS in sheep feed did not influence (P > 0.05) the voluntary water intake. The dietary DCS levels linearly affected (P < 0.05) the digestibility coefficients of the dry matter (DM), organic matter, neutral detergent fibre, ether extract, and non-fibrous carbohydrates. The DCS inclusion levels did not affect the nitrogen balance (P = 0.22), plasma concentrations of urea (P = 0.51), and glucose (P = 0.17).The feeding efficiency and rumination in g DM/h reduced linearly (P < 0.05) with the addition of DCS in the diet. Therefore, the addition of delinted cottonseed affects the intake, digestibility, and ingestive behaviour of confined sheep, and the addition of up to 80 g/kg delinted cottonseed in the diet of confined sheep is recommended.

Correction to: Cryptosporidium Spp. Contamination in Perna perna Mussels Destined for Human Consumption in Southeastern Rio de Janeiro, Brazil.

The original version of this article unfortunately contained a mistake.

Cryptosporidium spp. Contamination in Perna perna Mussels Destined for Human Consumption in Southeastern Rio de Janeiro, Brazil.

Cryptosporidium spp. has been recognized as an important pathogen. As bivalve mollusks are noted as potential sources of several pathogens due to their consumption as foodstuffs, the aim of this study was to investigate the occurrence of Cryptosporidium spp. oocysts in Perna perna mussels and in seawater samples from a mussel farm in Southeastern Brazil, where mussels are grown directly in the sea, attached to ropes. Oocysts were observed by microscopy and confirmed by an enzyme-linked immunosorbent assay. Oocysts were present in mussel gills and GI tracts, as well as in the seawater. Of the 100 females, 10% and 11% showed contaminated GI tracts and gills, respectively, while this rate was lower in males, at 5% and 8.9%. Oocysts were present in higher amounts in the GI tract compared to gills and water. Contamination of the study area is apparent, leading to public health risks. More in-depth studies are needed, including molecular investigations, to identify Cryptosporidium species in mussels, as well as the implementation of monitoring actions in animals destined for human consumption.

A proteomic approach to guarana seed and pericarp maturation.

Paullinia cupana Kunth var. sorbilis (Mart.) Ducke, the cultivated guarana plant, is native to the Amazon and has been valued for its medicinal, stimulant and energetic properties for centuries. The seeds are the main commercial product of the plant and the source of high amounts of purine alkaloids (caffeine and theobromine) and polyphenols (flavonoids, catechins, and tannins). Proteins involved in the development and maturation of guarana fruits in its native habitat are interesting issues for proteomics. This study presents the proteomic profile of the seed and pericarp of healthy guarana in different maturation stages. Protein contents were higher in the mature seed compared to other stages due to the accumulation of storage proteins - 11S globulins. Proteins selected for identification by mass spectrometry are mostly related to stress responses and defense and this is not unexpected for fast growing and differentiating reproductive tissues.

Interactions Among Different Devices and Electrical Stimulus on the Electroejaculation of Captive Agoutis (Dasyprocta leporina).

The interactions among different electroejaculation devices associated with serial or continuous stimuli were investigated to improve the efficiency of the electroejaculation for semen collection in agoutis. Ten sexually matured male Dasyprocta leporina were restrained by the intramuscular administration of xylazine-ketamine association. Each individual was randomly subjected to four electroejaculation protocols, by combining two devices (one presenting longitudinal electrodes emitting square waves and other presenting ring electrodes emitting sine waves) and two electrical stimuli protocols (serial or continuous). A total of 40 attempts for electroejaculation were conducted in agoutis, being 10 per treatment. The most efficient treatment in providing ejaculates containing sperm (p < 0.05) was that using and electroejaculator connected to a probe with ring electrodes and associated with serial stimuli (4/7; 57%). In spite of semen parameters obtained by sine waves were adequate for using the samples for assisted reproduction, higher values for sperm motility and functional membrane integrity were obtained in the use of the square wave, independently of the electric stimulation protocol used (p < 0.05). In conclusion, we verified that the use of a device presenting a probe with ring electrodes and emitting sine waves, associated with a serial stimuli protocol, improves the efficiency for semen obtaining by electroejaculation in adults D. leporina.

Isolation of Pantoea ananatis from sugarcane and characterization of its potential for plant growth promotion.

Each year, approximately 170 million metric tons of chemical fertilizer are consumed by global agriculture. Furthermore, some chemical fertilizers contain toxic by-products and their long-term use may contaminate groundwater, lakes, and rivers. The use of plant growth-promoting bacteria may be a cost-effective strategy for partially replacing conventional chemical fertilizers, and may become an integrated plant nutrient solution for sustainable crop production. The main direct bacteria-activated mechanisms of plant growth promotion are based on improvement of nutrient acquisition, siderophore biosynthesis, nitrogen fixation, and hormonal stimulation. The aim of this study was to isolate and identify bacteria with growth-promoting activities from sugarcane. We extracted the bacterial isolate SCB4789F-1 from sugarcane leaves and characterized it with regard to its profile of growth-promoting activities, including its ability to colonize Arabidopsis thaliana. Based on its biochemical characteristics and 16S rDNA sequence analysis, this isolate was identified as Pantoea ananatis. The bacteria were efficient at phosphate and zinc solubilization, and production of siderophores and indole-3-acetic acid in vitro. The isolate was characterized by Gram staining, resistance to antibiotics, and use of carbon sources. This is the first report on zinc solubilization in vitro by this bacterium, and on plant growth promotion following its inoculation into A. thaliana. The beneficial effects to plants of this bacterium justify future analysis of inoculation of economically relevant crops.

Morphometry, Morphology and Ultrastructure of Ring-tailed Coati Sperm (Nasua nasua Linnaeus, 1766).

The ring-tailed coati (Nasua nasua) is a procyonid whose population is in sharp decline. Therefore, studies are needed to better understand the reproduction of this animal. For this reason, this study aimed to evaluate the morphology, morphometry and sperm ultrastructure of ring-tailed coati sperm. Four captive adult males were used for this study. Slides stained with Bengal Rose were used for the morphometric and morphologic analyses. The length and width of the head were measured, as well as the length of the midpiece and tail and the total length of the sperm. Scanning electron microscopy and transmission electron microscopy were used for the ultrastructural analyses. The most obvious morphological abnormalities observed were coiled tails (6.1 ± 8.7%) and the lack of acrosomes (5.4 ± 4.4%). Regarding the morphometry, the measurements of the head (length × width), midpiece (length) and tail (length) were (mean ± SD) 6.2 ± 0.4 × 8.1 ± 0.6 µm, 14.1 ± 0.5 and 63.9 ± 4.1 µm, respectively, and the total length of the sperm was 86.1 ± 4.3 µm. Through electron microscopy, the presence of electron-lucent points in the nucleus and the presence of approximately 55 mitochondrial spirals in the midpiece were identified. The data obtained in this study provide detailed information on the sperm characteristics of coatis and may inform future research on germplasm conservation, both for this species and other threatened procyonids.

Method for obtaining high-resolution proteomic analysis from pericarps of guarana.

Guarana has great agricultural potential and is largely used therapeutically and in the production of non-alcoholic energy drinks. Genomic and proteomic studies are crucial to identify proteins that play central roles in the maintenance and viability of fruits, as well as to identify proteins related to the main metabolic pathways. However, the success of any protein analysis starts with the protein extract preparation, which needs to offer an extract that is free of contaminants. This study aimed to evaluate different extraction methods to obtain high-quantity and high-quality extracts that are compatible with analysis by 2-dimensional electrophoresis and tandem mass spectrometry protein identification. Three different methods were tested: trichloroacetic acid (TCA)/acetone, sodium dodecyl sulfate (SDS)/phenol, and polyvinylpolypyrrolidone (PVPP)/SDS/phenol. The extract obtained from the TCA/acetone precipitation presented low solubility and contamination with lipids and carbohydrates. On the other hand, the quality of the extract gradually improved after using phenol and PVPP/phenol, enabling a yield up to 2 mg/g macerated tissues and the detection of 457 spots by 2-dimensional electrophoresis. The effectiveness of the procedure used was validated by identification of 10 randomly selected proteins by mass spectrometry. The procedure described here can be a starting point for applications using tissues of other organs of guarana or tissues of species that are similar to guarana.

Comparison of different glycerol and egg yolk concentrations added to tris-based extender for the collared peccaries (Tayassu tajacu) semen freezing.

This study aimed to evaluate various concentrations of egg yolk (5, 10, or 20%) in combination with different concentrations of glycerol (3% or 6%) added to a Tris-based extender on the post-thaw characteristics of sperm obtained from Tayassu tajacu. For this purpose, semen from 10 sexually male mature collared peccaries was collected by electroejaculation and evaluated for sperm motility, vigour, viability, morphology and functional membrane integrity. The ejaculates were initially extended in Tris-fructose plus egg yolk (5%, 10% or 20%). After cooling, the semen was added to Tris-egg yolk plus glycerol (6% or 12%), resulting in a final concentration of 3% or 6% glycerol of the extender. Straws were frozen using liquid nitrogen and thawed in a water bath at 37°C for 30 s. The frozen-thawed semen was evaluated as reported for fresh semen. After thawing, a significant decrease was verified for sperm motility and vigour, for all the samples in comparison with fresh semen. However, no differences were evidenced among treatments for any sperm characteristics evaluated (p > 0.05), except for the combination between 10% egg yolk and 6% glycerol, which provided the worst preservation of functional membrane integrity (p < 0.05). The interactions between higher concentrations of egg yolk (20%) and glycerol (6%) and also between lower concentrations of the same substances (5% egg yolk and 3% glycerol) added to the Tris-based extender negatively affected the preservation of the normal sperm morphology after thawing (p < 0.05). In conclusion, the use of Tris-based extender added to 10% or 20% egg yolk plus 3% glycerol is recommended for effective sperm cryopreservation in collared peccaries.

Extract of Cimicifuga racemosa (L.) Nutt protects ovarian follicle reserve of mice against in vitro deleterious effects of dexamethasone.

The present study aims to investigate if Cimicifuga racemosa (L.) Nutt extract (CIMI) reduces deleterious effects of dexamethasone (DEXA) in ovaries cultured in vitro. Mouse ovaries were collected and cultured in DMEM+ only or supplemented with 5 ng/mL of CIMI, or 4 ng/mL DEXA, or both CIMI and DEXA. The ovaries were cultured at 37.5°C in 5% CO2 for 6 days. Ovarian morphology, follicular ultrastructure, and the levels of mRNA for Bax, Bcl-2, and Caspase-3 were evaluated. The results showed that DEXA reduced the percentage of morphologically normal follicles, while CIMI prevented the deleterious effects caused by DEXA. In addition, DEXA negatively affected the stromal cellular density, while CIMI prevented these adverse effects. Ovaries cultured with DEXA and CIMI showed similar levels of mRNA for Bax, Bcl-2, and Caspase-3 compared to those cultured in control medium, while ovaries cultured with DEXA had increased expression of the above genes. Additionally, the ultrastructure of the ovaries cultured with CIMI was well preserved. Thus, the extract of CIMI was able to prevent the deleterious effects caused by DEXA on cultured mouse ovaries.

Immunolocalization of melatonin receptors in bovine ovarian follicles and in vitro effects of melatonin on growth, viability and gene expression in secondary follicles.

This study aims to investigate the (1) expression of melatonin receptors types 1A/B (MTNR1A/B) in bovine ovaries and (2) the in vitro effects of melatonin on secondary follicle development, antrum formation, viability, and expression of messenger ribonucleic acid (mRNA) for superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase-1 (GPX1) and peroxiredoxin 6 (PRDX6). The expression of MTNR1A/B in bovine ovarian follicles was demonstrated by immunohistochemistry. To choose the most effective concentration of melatonin on follicular growth and viability, isolated secondary follicles were cultured individually at 38.5°C, with 5% CO2 in air, for 18 d in TCM-199+ alone or supplemented with 10-11, 10-9, 10-7 or 10-5 M melatonin. Then, melatonin receptor antagonist, luzindole, was tested to further evaluate the mechanisms of actions of melatonin, that is, the follicles were cultured in control medium alone or supplemented with 10-7 M melatonin, 10 µM luzindole and both 10-7 M melatonin and 10 µM luzindole. Follicular growth, morphology and antrum formation were evaluated at days 6, 12 and 18. At the end of culture, viability of secondary follicles was analyzed by calcein-AM and ethidium homodimer-1, and the relative levels of mRNA for SOD, CAT, GPX1 and PRDX6 were evaluated by real time polymerase chain reaction. Immunohistochemistry results showed expression of MTNR1A/B in oocyte and granulosa cells of primordial, primary, secondary and antral follicles. Secondary follicles cultured in medium supplemented with melatonin at different concentrations had well preserved follicles after 18 d of culture. Furthermore, follicles cultured in presence of 10-7 M melatonin presented significantly higher diameters than those cultured in other treatments. The presence of melatonin receptor antagonist, luzindole, blocked the effects of melatonin on follicular growth and viability. In addition, follicles cultured in medium containing only melatonin had significantly higher rates of antrum formation. Follicles cultured in medium containing only melatonin had higher relative levels of mRNA for CAT, SOD and PRDX-6 than those cultured with both melatonin and luzindole. Follicles cultured with luzindole only or both melatonin and luzindole had lower relative levels of mRNA for PRDX6 and GPX1 than those cultured control medium. In conclusion, melatonin promotes growth of bovine secondary follicles through its membrane-coupled receptors, while luzindole blocks the effects of melatonin on follicle growth and reduces the expression of antioxidant enzymes in cultured follicles.

Looking for reference genes for real-time quantitative PCR experiments in Rhodnius prolixus (Hemiptera: Reduviidae).

Quantitative real-time PCR (qPCR) has become one of the most used techniques to measure gene expression. However, normalization of gene expression data against reference genes is essential, although these are usually used without any kind of validation. The expression of seven genes was compared in organs of Rhodnius prolixus under diverse conditions, using published software to test gene expression stability. Rp18S and elongation factor 1 (RpEF -1) were the most reliable genes for normalization in qPCR when gene expression in different organs was compared. Moreover, both genes were found to be the best references when transcript levels were compared in the posterior midgut of insects infected with Trypanosoma cruzi. Rp18S was also the best reference gene in the fat bodies of unfed and fed insects. By contrast, RpEF-1 was found to be the best reference gene for comparison between posterior midguts, and RpMIP or RpActin should be used to compare gene expression in the ovaries. Although Rp18S is indicated here as the best reference in most cases, reports from the literature show that it is difficult to find an optimum reference gene. Nevertheless, validation of candidate genes to be taken as references is important when new experimental conditions are tested to avoid incorrect data interpretation.

Formulating octyl methoxycinnamate in hybrid lipid-silica nanoparticles: An innovative approach for UV skin protection.

Sunscreens have been employed on daily skin care for centuries. Their role in protecting the skin from sun damage, avoiding accelerated photoaging and even limiting the risk of development of skin cancer is unquestionable. Although several chemical and physical filters are approved as sunscreens for human use, their safety profile is dependent on their concentration in the formulation which governs their acceptance by the regulatory agencies. A strategic delivery of such molecules should provide a UV protection and limit the skin penetration. Solid lipid nanoparticles (SLN) and nanostructured lipid carriers (NLC) may offer an alternative approach to achieve a synergistic effect on the UV protection when loaded with sunscreens as particles themselves also have a UV light scattering effect. Besides, the lipid character of SLN and NLC improves the encapsulation of lipophilic compounds, with enhanced loading capacity. Silica nanoparticles have also been employed in sunscreen formulations. Due to the formed sol-gel complexes, which covalently entrap sunscreen molecules, a controlled release is also achieved. In the present work, we have developed a new sunscreen formulation composed of hybrid SLN-Silica particles loaded with octyl methoxycinnamate (Parsol®MCX), and their further incorporation into a hydrogel for skin administration. Hybrid SLN-silica particles of 210.0 ± 3.341 nm of mean size, polydispersity below 0.3, zeta potential of ca. |7| mV, loading capacity of 19.9% and encapsulation efficiency of 98.3% have been produced. Despite the slight negative surface charge, the developed hybrid nanoparticles remained physicochemically stable over the study period. Turbiscan transmission profiles confirmed the colloidal stability of the formulations under stress conditions. The texture profile analysis of Parsol-SLN and Parsol-SLN-Si revealed semi-solid properties (e.g. adhesiveness, hardness, cohesiveness, springiness, gumminess, chewiness, resilience) suitable for topical application, together with the bioadhesiveness in the skin of pig ears. The non-irritation profile of the hybrid nanoparticles before and after dispersion into Carbopol hydrogels was confirmed by HET-CAM test.

Effects of epidermal growth factor and progesterone on development, ultrastructure and gene expression of bovine secondary follicles cultured in vitro.

The aims of this study were to investigate the effects of epidermal growth factor (EGF) and progesterone on the development, viability and the gene expression of bovine secondary follicle culture in vitro for 18 days. Secondary follicles (∼0.2 mm) were isolated from ovarian cortex and individually cultured at 38.5 °C, with 5% CO2 in air, for 18 days, in TCM-199+ (n = 63) alone (control medium) or supplemented with 10 ng/mL progesterone (n = 64), 10 ng/mL EGF (n = 61) or both EGF and progesterone (n = 66). The effects of these treatments on growth, antrum formation, viability, ultrastructure and mRNA levels for GDF-9, c-MOS, H1foo and cyclin B1 were evaluated, significantly different (p < 0.05). The results showed that there was a progressive increase in follicular diameter in all treatments, but only follicles cultured in medium supplemented with EGF had increased significantly in diameter when compared to follicles cultured in the control medium at the end of the culture period, significantly different (p < 0.05). A positive interaction between EGF and progesterone was not observed. In addition, the presence of EGF, progesterone or both in culture medium did not influence the rate of follicle survival and antrum formation. However, the presence of only progesterone in cultured medium increased the expression of mRNAs for GDF9 and cyclin B1 in oocytes. EGF also significantly increased the levels of mRNAs for cMOS and GDF9 when compared to follicles cultured in control medium. Ultrastructural analyzes showed that cultured follicles in all treatments maintained the integrity of granulosa cells. In conclusion, the EGF promotes the development of secondary follicles cultured in vitro for 18 days and increases the expression of cMOS and GDF9, while progesterone alone or in association with EGF have not a positive effect on follicular growth. However, progesterone increases the expression of GDF9 and cyclin B1 in oocytes.

Environmental effects on collared peccaries (Pecari tajacu) serum testosterone, testicular morphology, and semen quality in the Caatinga biome.

The objective of the study was to understand the influence of climatic variations in a semiarid environment on serum testosterone, testicular morphology and semen quality in collared peccaries (Pecari tajacu). Reproductive metrics (semen quality, testicular morphometry and testosterone serum profiles) of 10 mature males were measured monthly for 18 months. Meteorological data (rainfall, air temperature, relative humidity, wind speed and radiant heat load) also were recorded during the same period. Rainfall regimes were classified in different classes (Class 1: months with no rain; Class 2: months with up to 50 mm of rain; and Class 3: months with >50 mm of rain). Among rainfall classes, average air temperature (°C) and relative humidity (%) were different. Climatic changes between rainfall classes did not lead to overall variations of testicular size, testosterone production, and semen metrics. However, relative humidity recorded before semen collection (one day, one week, or over 51-55 days) was positively correlated (P < 0.05) with semen motility metrics (total motility, beat cross frequency and straightness) and sperm subpopulations (medium and static sperm), as well as with volume. Negative correlations (P < 0.05) were revealed between air temperature and the same semen motility patterns and volume. Additionally, radiant head load measured on the day of semen collection negatively influenced (P < 0.05) sperm straightness. This study demonstrates for the first time that no seasonal changes could be detected overt the 18-month period on the serum testosterone, testicular morphology and semen quality of collared peccaries raised in the Caatinga biome; however, it is expected that long term environmental changes will influence the reproductive physiology of species leaving in that habitat.

Loxosceles venom-induced cytokine activation, hemolysis, and acute kidney injury.

Herein, we describe a confirmed case of Loxosceles spider bite that illustrates the critical complications seen in loxoscelism, including skin necrosis, rhabdomyolysis, hemolysis, coagulopathy, acute kidney failure, and electrolyte disorders. Upon initial assessment, laboratory studies revealed the following: the white blood cell count was 29,400 WBCs/mm(3), hemoglobin was 9.2g/dL, and the platelet count was 218,000 cells/mm(3). Coagulation studies revealed the following: international normalized ratio, 1.83; activated partial-thromboplastin time, 62 s; D-dimer, 600 ng/mL (normal range <500 ng/mL); free protein S, 37% (normal range=64-114%); protein C, negative; and antithrombin III, negative. Various serum levels were abnormal: urea, 110 mg/dL; creatinine, 3.1mg/dL; indirect bilirubin, 3.8 mg/dL; creatine kinase, 1631 U/L; lactate dehydrogenase, 6591 U/L; potassium 6.2 mmol/L. Urine tests were positive for hemoglobin and bilirubin. In addition, concentrations of interleukin-6 and tumor necrosis factor-alpha were notably elevated in the serum. In conclusion, physicians must be alert to the possibility of loxoscelism when a patient presents with the clinical and laboratory findings described above, especially if the patient resides in an endemic area. Advances in our understanding of multiple pathways and mediators that orchestrate the response to Loxosceles venom might reveal new possibilities for the management of loxoscelism.

Embryo production in superovulated goats treated with insulin before or after mating or by continuous propylene glycol supplementation.

Seventeen adult and cyclic Moxoto goats were synchronized using 60 mg MPA vaginal sponge for 11 days and 50 mug cloprostenol, 48 h before sponge removal, and superovulated with 120 mg pFSH i.m. in decreasing doses at 12 h intervals for three consecutive days. In seven goats, 0.2 IU/kg BW/day of long acting insulin was subcutaneously injected at same time as pFSH, and in the other five goats, the same dose of insulin was injected for three consecutive days starting 24 h after mating. Finally, five goats were supplemented with an oral dose of 80 ml/goat/day of propylene glycol continuously during the experiment. The animals were flushed at 7 days after mating and the embryos were classified based on International Embryo Transfer Society criteria. Blood samples were collected every 3 days for insulin assay. Administration of insulin raised the insulin levels of the goats (p < 0.05), whereas in the group treated with propylene glycol, insulin rate was different only between FSH treatment and after mating (p < 0.05). Similar rates of recovery for total (80.05 +/- 9.78%) or transferable structures (61.03 +/- 15.13%) were obtained. Treatment was not influenced (p > 0.05) by responsiveness to superovulation, which averaged 64%. By contrast, insulin treatments were shown to increase the number of embryos considered excellent with respect to goats supplemented with propylene glycol (p < 0.05). When insulin was given before mating, a strong relationship (r = 0. 90) (p < 0.05) between number of transferable embryo and ovulations was observed in the animals. In conclusion, superovulated goats treated with low doses of exogenous insulin resulted in an enhancement in embryo quality, which was related to changes in circulating insulin concentrations.