Progesterone production in mares and echographic evaluation of the corpora lutea formed after follicular aspiration

Ultrasound‐guided follicular aspiration was performed in 26 Criollo crossbred mares, followed by the evaluation of ultrasonographic images of the Corpus luteum (CL) that was formed after puncture of follicles of different diameters (Group 25–29 mm; Group 30–35 mm and Group >35 mm). Serum progesterone (P₄) concentrations were measured to determine CL function. The size of the CL was measured and the CL was classified based on the following echoscore: 1– anechoic tissue; 2– poorly defined luteal structure with low echogenicity; 3– echogenicity analogous to a luteal structure. The proportion of aspirated follicles that formed a functional CL (based on P₄ concentration) 8 days after aspiration was 57.1% (4/7; CL size 25–29 mm), 75.0% (6/8; CL size 30–35 mm) and 72.7% (8/11; CL size >35 mm), respectively (p > 0.05). The echographic scores of aspirated follicles (indicating the presence or absence of a CL) were consistent with serum P₄ concentrations (p < 0.0001). Of 26 aspirations, 18 resulted in luteal function confirmed by increased progesterone concentrations ([P₄] > 1.0 ng/ml); 17 of these mares (94.4%) had an echoscore (2–3) compatible with luteinization (p = 0.0372). Eight days after aspiration, serum [P₄] > 2.0 ng/ml was associated with high (p = 0.0056) CL echoscore (3) in 15 of 17 mares (88.2%). The echoscore used in this study was valuable as a screening test to detect the presence of a functional CL after aspiration. An echoscore of 3 served as a practical and efficient method to confirm luteinization.     

Sheep fed with banana leaf hay reduce ruminal protozoa population

A ciliate protozoa suppression can reduce methane production increasing the energy efficiency utilization by ruminants. The physicochemical characteristics of rumen fluid and the profile of the rumen protozoa populations were evaluated for sheep fed banana leaf hay in replacement of the Cynodon dactylon cv. vaqueiro hay. A total of 30 male sheep were raised in intensive system during 15 days of adaptation and 63 days of experimental period. The animals were distributed in a completely randomized design that included six replicates of five treatments with replacement levels (0, 25, 50, 75, and 100%) of the grass vaquero for the banana leaf hay. Samples of fluid were collected directly from the rumen with sterile catheters. Color, odor, viscosity, and the methylene blue reduction potential (MBRP) were evaluated and pH estimated using a digital potentiometer. After decimal dilutions, counts of genus protozoa were performed in Sedgewick Rafter chambers. The averages of pH, MBRP, color, odor, and viscosity were not influenced by the inclusion of the banana leaf hay. However, the total number of protozoa and Entodinium spp. population significantly decreased at 75 and 100% inclusions of banana leaf hay as roughage.     

Effects of α-tocopherol and Ascorbic Acid on Equine Semen Quality after Cryopreservation

This study investigated the effects of ascorbic acid and α-tocopherol supplementation on semen quality parameters of equine thawed-frozen semen. Semen was divided in seven different treatments in a final concentration of 100 × 10⁶ sperm/mL by using Gent extender containing no supplements (control) and the following supplements with three different concentrations: α-tocopherol (0.5, 1, and 2 mM) and ascorbic acid (0.45, 0.9, and 1.8 g/L). After thawing, all samples were maintained at 37°C, while analyses were performed at 0, 60, and 120 minutes. Evaluation of viability and acrosome status (using Pisum sativum agglutinin conjugated to fluorescein isothiocyanate and propidium iodide), mitochondrial membrane potential (5,5′,6,6′-tetrachloro-1,1′,3,3′tetraethylbenzimidazolyl carbocyanine iodine [JC-1]), membrane lipid peroxidation (LPO; C₁₁-BODIPY^581/591), and stability of the plasmatic membrane (merocyanine 540 and Yo-Pro-1) of each sample was determined by flow cytometry. Relative to the control group, supplementation with α-tocopherol improved (P ≤ .05) postthaw membrane LPO, yet the higher concentrations of ascorbic acid (0.9 and 1.8 g/L, respectively) showed a negative effect on membrane LPO. Neither antioxidant significantly increased (P > .05) the acrosome integrity and mitochondrial membrane potential of frozen-thawed spermatozoa, although supplementation with α-tocopherol and ascorbic acid (0.9 and 1.8 g/L, respectively) had a positive effect on membrane integrity and stability (P ≤ .05). For all semen parameters, the lower concentration of ascorbic acid (0.45 g/L) did not show significant differences (P > .05) compared with the control. In conclusion, α-tocopherol seems to be an efficient antioxidant for reducing the oxidative stress provoked by cryopreservation, decreasing lipid peroxidation on equine spermatozoa.     

Effect of Oestrous Cycle on the Oxidative Burst Activity of Blood Polymorphonuclear Leucocytes in Cows

Blood polymorphonuclear leucocyte (PMN) oxidative burst activity, plasma cortisol levels, and the total and differential white blood cells counts (WBC) of six cycled dairy cows were evaluated for a period of 24 days, three times a week; on Mondays, Wednesdays and Fridays. The PMN oxidative burst was indirectly evaluated by flow cytometry, measuring the intracellular oxidation of 2′,7′‐dichlorofluorescein diacetate to 2′,7′ dichlorofluorescein (DCF) by H₂O₂‐production. Results are pre‐sented as the mean fluorescence intensity (MFI) of DCF. Cow’s oestrous cycle was evaluated by following the plasma progesterone levels using a radioimmunoassay method. Levels of cortisol in the plasma were measured using a fluorimetric method. The oxidative burst activity of PMN, represented a maximum value (MFI = 117.6 ± 7.4) during the oestrous period. A fall was then observed, in which a steady state was observed during the lutheinic phase of the oestrous cycle, reaching the minimum value [MFI = 73.2 ± 11.2 (p ≤ 0.01)] on the days +8, +9 and +10. No significant variations were observed in the levels of cortisol, or in total and differential WBC, during the whole period. Nevertheless, as far as cortisol levels were concerned, a trend analogous to that of the oxidative burst activity was observed. Our results demonstrated that the oestrous cycle might influence directly, or indirectly, the immune system of cows, by altering the oxidative burst of PMN.     

Immunolocalization and pathological alterations following Strongyloides venezuelensis infection in the lungs and the intestine of MHC class I or II deficient mice

The present study, investigated the mechanisms involved in the immune responses of Major Histocompatibility Complex class I or class II knockout mice, following Strongyloides venezuelensis infection. Wild-type C57BL/6 (WT), MHC II(-/-) and MHC I(-/-) mice were individually inoculated with 3000 larvae (L3) of S. venezuelensis and sacrificed on days 1, 3, 5, 8, 13 and 21 post-infection (p.i.). Samples of blood, lungs and small intestines were collected. The tissue samples were stained with hematoxylin-eosin for the pathological analysis. The presence of the parasite was demonstrated by immunoperoxidase analysis. MHC II(-/-) mice presented a significantly higher number of adult worms recovered from the small intestine on day 5p.i. and presented elevated numbers of eggs in the feces. The infection by S. venezuelensis was completely eliminated 13 days after infection in WT as well as in MHC I(-/-) mice. In MHC II(-/-) mice, eggs and adult worms were still found on day 21 p.i., however, there was a significant reduction in their numbers. In the lung, the parasite was observed in MHC I(-/-) on day 1 p.i. and in MHC II(-/-) mice on days 1 and 5 p.i. In the small intestine of WT mice, a larger number of parasites were observed on day 8 p.i. and their absence was observed after day 13 p.i. Through immunohistochemistry analysis, the parasite was detected in the duodenum of WT on days 5 and 8 p.i., and in knockout mice on days 5, 8 and 13 p.i.; as well as in posterior portions of the small intestine in MHC I(-/-) and MHC II(-/-) on day 13 p.i., a finding which was not observed in WT mice. We concluded that immunohistochemistry analysis contributed to a more adequate understanding of the parasite localization in immunodeficient hosts and that the findings aid in the interpretation of immunopathogenesis in Strongyloides infection.     

Prevalence and risk factors for anti-Toxoplasma gondii antibodies in goats of the Seridó Oriental microregion, Rio Grande do Norte state, Northeast region of Brazil

The prevalence and risk factors for anti-Toxoplasma gondii antibodies were investigated in goats of the Seridó Oriental microregion, Rio Grande do Norte state, Northeast region of Brazil. Three hundred and sixty-six blood samples from goats collected by jugular venopuncture were used. For the serologic diagnosis of Toxoplasma gondii infection, the indirect fluorescent-antibody test (IFAT) with cut-off value 1:64 was carried out. The prevalence of anti-T. gondii antibodies was 30.6% [95% CI=25.9-35.6%] with titers ranging from 1:64 to 1:16,384. The multivariate logistic regression analysis showed that the risk factors associated to anti-T. gondii antibodies were presence of cats in the herd, extensive/semi-intensive management systems and lack of mineral supplementation.     

Estimation of genetic parameters for mature weight in Angus cattle

The aim of this study was to estimate genetic parameters for BW of Angus cattle up to 5 yr of age and to discuss options for including mature weight (MW) in their genetic evaluation. Data were obtained from the American Angus Association. Only records from herds with at least 500 animals and with >10% of animals with BW at ≥ 2 yr of age were considered. Traits were weaning weight (WW, n = 81,525), yearling weight (YW, n = 62,721), and BW measured from 2 to 5 yr of age (MW2, n = 15,927; MW3, n = 12,404; MW4, n = 9,805; MW5, n = 7,546). Genetic parameters were estimated using an AIREML algorithm with a multiple-trait animal model. Fixed effects were contemporary group and departure of the actual age from standard age (205, 365, 730, 1,095, 1,460, and 1,825 d of age for WW, YW, MW2, MW3, MW4, and MW5, respectively). Random effects were animal direct additive genetic, maternal additive genetic, maternal permanent environment, and residual. Estimates of direct genetic variances (kg(2)) were 298 ± 71.8, 563 ± 15.1, 925 ± 52.1, 1,221 ± 65.8, 1,406 ± 80.4, and 1,402 ± 66.9; maternal genetic variances were 167 ± 4.8, 153 ± 6.1, 123 ± 9.1, 136 ± 12.25, 167 ± 18.0, and 110 ± 14.0; maternal permanent environment variances were 124 ± 2.9, 120 ± 4.3, 61 ± 7.5, 69 ± 11.9, 103 ± 15.9, and 134 ± 35.2; and residual variances were 258 ± 3.8, 608 ± 8.6, 829 ± 34.2, 1,016 ± 38.8, 1,017 ± 52.1, and 1,202 ± 63.22 for WW, YW, MW2, MW3, MW4, and MW5, respectively. The direct genetic correlation between WW and YW was 0.84 ± 0.14 and between WW and MW ranged from 0.66 ± 0.06 (WW and MW4) to 0.72 ± 0.11 (WW and MW2). Direct genetic correlations ranged from 0.77 ± 0.08 (YW and MW5) to 0.85 ± 0.07 (YW and MW2) between YW and MW, and they were ≥ 0.95 among MW2, MW3, MW4, and MW5. Maternal genetic correlations between WW and YW and MW ranged from 0.52 ± 0.05 (WW and MW4) to 0.95 ± 0.07 (WW and YW), and among MW they ranged from 0.54 ± 0.14 (MW4 and MW5) to 0.94 ± 0.07 (MW2 and MW3). Genetic correlations suggest that a genetic evaluation for MW may be MW2-based and that including BW from older ages could be accomplished by adjusting records to the scale of MW2.     

Quantitative and descriptive histological aspects of jaguar (Panthera onca Linnaeus, 1758) ear skin as a step towards formation of biobanks

Skin of mammals vulnerable to extinction, such as the jaguar, is used as a source of material in conservation strategies. The composition of skin is not uniform among species, and the ability to distinguish similarities in skin morphology in animal groups is fundamental in the application of skin tissue for use in biobanks. The aim of our study was to evaluate the structure, composition and capacity for culture of ear skin from the yellow and black jaguars. Both qualitative and quantitative methods were used, focusing on skin thickness, cell quantification and distribution, collagen density, proliferative activity and viability. Histomorphometrical study of the skin showed a total thickness of 273.2 and 274.6 µm for the yellow and black jaguars, respectively. Melanocytes and fibroblasts were, respectively, 9.7 and 23.0 for the yellow jaguar and 11.3 and 26.8 for the black jaguar. A collagen density of 67.0% and 49.0% was observed for yellow and black jaguars, respectively. Both animals presented a proliferative activity varying between 1.20 and 1.30. All tissues could promote cellular detachment, reaching subconfluence in 10–15 days. This kind of information from histomorphometrical features and cell cultures can be essential for a more targeted application of ear skin cryopreservation in this species, as such information will enable understanding the action of substances on tissues during the conservation process.     

Fatty acid composition of intramuscular fat from Nellore steers fed dry or high moisture corn and calcium salts of fatty acids

The purpose of this work was to evaluate the fatty acid composition of the Longissimus muscle from carcasses of Nellore steers fed diets with calcium salts of fatty acids (CSFA) and high moisture corn. Forty eight steers were fed during 70 days four diets containing dry corn (DC), high moisture corn (HM), dry corn plus CSFA (DC-CSFA) or high moisture corn plus CSFA (HM-CSFA). Fatty acid composition of the Longissimus muscle was determined by gas chromatography. Corn type had no effect on the ether extract percentage and in the content of the majority of the fatty acids, although steers fed HMC showed higher levels of polyunsaturated fatty acids and polyunsaturated/saturated ratio. Feeding CSFA increased ether extract percentage but had no effect on total of saturated, unsaturated and saturated: unsaturated ratio. Both high moisture corn and calcium salts of fatty acids increased CLA (cis9, trans11) and total CLA concentrations in intramuscular fat.     

Different isolates from Leishmania braziliensis complex induce distinct histopathological features in a murine model of infection

The aim of this study was to evaluate the histopathological features in tissues of mice infected by human isolates (I, II, and III) or the reference M2903 strain of Leishmania braziliensis complex. BALB/c and C57Bl/6 mice were infected in the hind footpad with 10⁶ stationary-phase promastigotes of L. braziliensis complex. The evolution of lesions was observed for 10 weeks and the animals were then euthanized and liver, spleen and popliteal lymph nodes were collected. Tissues were stained with hematoxylin and eosin and analyzed by immunohistochemistry assay. Increased thickness of infected footpads was observed in all animals, lesions were nodular and non-ulcerated. Mice infected with isolate I presented inflammatory infiltrates consisting predominantly of mononuclear cells in all tissues examined, and also a great number of megakaryocytes, compared with other isolates. Infection with isolate II led to an infected footpad enlargement not seen in other isolates. In addition, mononuclear infiltrates in the liver and hemosiderin in spleen were noted. Conversely, mice infected with either isolate III or M2903 strain only showed an increased number of megakaryocytes in spleen. All tissues examined had detectable amastigote forms of Leishmania by immunohistochemistry in all groups. Taking together, our results showed an unforeseen behavior of different isolates of L. braziliensis complex that led to diverse pathological findings.