Ultrastructure of sheep primordial follicles cultured in the presence of indol acetic acid, EGF, and FSH

The aim of this study was to investigate the ultrastructural characteristics of primordial follicles after culturing of sheep ovarian cortical slices in the presence of indol acetic acid (IAA), Epidermal Growth Factor (EGF), and FSH. To evaluate ultrastructure of primordial follicles cultured in MEM (control) or in MEM containing IAA, EGF, and FSH, fragments of cultured tissue were processes for transmission electron microscopy. Except in the control, primordial follicles cultured in supplemented media for 6d were ultrastructurally normal. They had oocyte with intact nucleus and the cytoplasm contained heterogeneous-sized lipid droplets and numerous round or elongated mitochondria with intact parallel cristae were observed. Rough endoplasmic reticulum (RER) was rarely found. The granulosa cells cytoplasm contained a great number of mitochondria and abundant RER. In conclusion, the presence of IAA, EGF, and FSH helped to maintain ultrastructural integrity of sheep primordial follicles cultured in vitro. © 2011 Evelyn Rabelo Andrade et al.

Effect of percutaneous transthoracic lung biopsy on oxidative metabolism in sheep

This study aimed to assess the effect of percutaneous transthoracic lung biopsy on the oxidative metabolism of sheep by measuring the oxidative stress markers of superoxide dismutase (SOD), total glutathione (GSH-t), peroxidase (GSH-Px) and thiobarbituric acid reactive substances (TBARS) in the red cells of these animals. Blood samples were collected from 20 clinically healthy sheep prior to, and 30 min after, percutaneous transthoracic lung biopsy. After biopsy, there was a significant decrease (p < 0.05) in SOD and GSH-Px activity, with no significant change (p ≥ 0.05) in GSH-t and TBARS concentrations. These results showed that percutaneous transthoracic lung biopsy did not significantly affect the oxidative metabolism of sheep 30 min after the procedure, which may be used widely in this species without causing serious tissue damage. © 2012. The Authors.

Supplementation of diets for Santa Ines sheep with organic and inorganic zinc sources

This research was conducted with objective to evaluate the effect of different zinc (Zn) sources and doses in the diet for Santa Ines sheep. Forty lambs at weaning, with 18.4 kg of body weight were supplemented with three different sources of zinc (zinc oxide (ZnO), zinc amino acid and zinc proteinate) and three doses of zinc (200, 400 and 600 mg/kg DM) added to the basal diet. At every 28 days, animals were weighted and blood samples were collected for analyses of zinc (Zn), alkaline phosphatase and immunoglobulin G (IgG) and M (IgM). At the end of experiment, liver samples were collected for determination of the hepatic zinc levels. Zinc was analyzed with atomic absorption spectrophotometer, while phosphatase alkaline and immunoglobulins G and M were analyzed using Laborlab and Bioclin kits, respectively. There was no effect of diets on phosphatase alkaline levels and hepatic zinc, but there was difference in the plasmatic zinc levels and IgG and IgM levels. Based on the accumulation of hepatic zinc, the estimate of the zinc bioavailability, through the regression equation, showed that supplementation with organic and inorganic sources of zinc did not differ in the diet of Santa Ines sheep. © 2012 Sociedade Brasileira de Zootecnia.

Effect of roscovitine and cycloheximide on ultrastructure of sheep oocytes

It is believed the temporary meiosis arrest with roscovitine or cycloheximide may improve the in vitro developmental competence of oocytes in different animal species. However, little is known about the effects of these inhibitors on ultrastructure of ovines cumulus-oocyte complexes (COCs). The aim of this study was to evaluate the progression of cytoplasmic maturation and the ultrastructural changes in sheep COCs exposed to roscovitine or cycloheximide, at acceptable concentrations. COCs were in vitro cultured for 24. h in maturation medium (control group) containing 100 μM roscovitine or 1 μg/mL cycloheximide (treatment groups). After this time, some COCs were cultured for further 22. h in inhibitor-free medium. The ultrastructure organization of COCs was evaluated by transmission electron microscopy before (immature group) and after in vitro culture for 24 and 46. h. As expected, signs of immaturity and maturity were observed in immature and control groups, respectively. In treatment with roscovitine, there were cumulus cells degeneration, swelling of mitochondrias, few cortical granules and many vesicles with electron-dense material. However, in cycloheximide treatment there were not signs of degeneration or cellular senescence. Metabolic units and mitochondrial pleomorphism were found in all experimental groups. These evidences demonstrate that roscovitine promoted irreversible ultrastructural changes while cycloheximide did not affect the cytoplasmic maturation. However, the implications on embryo development are still unclear. © 2012 Elsevier B.V.

Supplementation with protected fat to manage gastro-intestinal nematode infections in Santa Ines sheep

This experiment aimed to determine the influence of supplementation with protected fat on nematode infections in periparturient sheep or sheep in their final stage of pregnancy. Fifty Santa Ines ewes received 200 g of concentrate/animal/day, mineral salt and water and were grouped according to their weight, hematocrit and the number of eggs per gram of feces (EPG). Two diets were used: a control treatment and a treatment consisting of supplementation with 30 g/animal/day of protected fat in the concentrate. Three monthly evaluations were performed over a total of 84 days of testing. The variables analyzed were weight, body condition, EPG and coprocultures; blood tests were performed for the determination of packed cell volume, hemoglobin concentration and total plasma protein, and leukograms and eosinophil counts were performed. For weight and hemoglobin concentrations, interactions were observed between diet and collection date (P<0.05); body condition, total plasma protein, packed cell volume and total leukocytes did not differ statistically among treatments (P> 0. 05) but did differ by collection day (P <0.05). The EPG and eosinophil counts did not differ statistically by either diet or collection date (P>0.05). The genus Haemonchus was predominant, followed by the genera Cooperia, Trichostrongylus and Oesophagostomum. The protected fat did not decrease EPG and did not improve the blood parameters of infected sheep.

Sexual transmission of Toxoplasma gondii in sheep

Male sheep of reproductive age were distributed into three groups: GI, a sheep inoculated (oral) with 2.0×105 oocysts of the P strain of Toxoplasma gondii; GII, a sheep infected (subcutaneous) with 1.0×106 tachyzoites of the RH strain of T. gondii; and GIII, a sheep kept as a control (not infected). After the inoculation of the males, 12 breeding ewes, which were not pregnant and which were serologically negative for reproductive diseases (particularly toxoplasmosis), were distributed into three groups, synchronized, and subsequently exposed to natural mating with previously inoculated males. The distribution was as follows: five ewes that underwent natural mating with the GI male, five ewes that were exposed to natural mating with the GII male, and two ewes that were mated with the non-infected male (control). Serum samples of all the ewes were collected on days -30, -14, -7, -1, and 0 (days before natural mating) and on days 1, 3, 5, 7, 11, 14, and weekly until birth; the presence of serum antibodies against T. gondii was assessed by IFAT. Using a bioassay and PCR, T. gondii was isolated from the semen of the infected reproducing sheep before mating. Following natural mating, 5 of the 12 females displayed antibodies specific for T. gondii; of these animals, two of the ewes underwent natural mating with the male inoculated with oocysts (GI) and three with the male infected with tachyzoites (GII). One of the females that displayed antibodies specific to this coccidian and that underwent natural mating with the GII sheep had a macerated fetus on the 70th day following coverage. Using a bioassay after the birth, it was possible to isolate T. gondii from samples of the pool of tissues from the five females that seroconverted after natural mating and from their respective lambs. Using PCR, the DNA of T. gondii was isolated from the pool of tissues from one and two females exposed to natural mating with the reproductive males infected with the oocysts and tachyzoites, respectively. Using this technique, it was also possible to diagnose the presence of the parasite in the pool of tissues from the lambs of one female that underwent natural mating with the male sheep infected with oocysts. These results demonstrated the sexual transmission of T. gondii in the sheep species with consequent vertical transmission to their lambs. © 2013 Elsevier B.V.

Morphology and biometric of the vesicular and bulbourethral glands in castrated and non-castrated Santa Ines breed sheep

Introduction: The purpose of this paper is to determine the topography, biometry and light microscopy image of the vesicular and bulbourethral glands in order to analyze morphologic features of the accessory sexual glands in castrated and non-castrated animals. Materials and Methods: The morphology of the accessory sexual glands was investigated in 14 adult Santa Ines breed sheep, weighing 32 kg, on average. Six of them were castrated, and eight, non-castrated. For macroscopic study, the description of these two glands was carried out, as well as dissection and biometry study. Moreover, weight, length, height and width measurements were evaluated. For histological analysis, the vesicular and bulbourethral glands were sampled. Results: The topography of the reproductive glands was similar to bovine species. However, lower macroscopic measurements (p < 0,05) in the glands of the castrated sheep were evidenced when compared with the non-castrated ones. Characteristics such as shape of the glands, composition of the layer mucosa, the lamina propria, muscular, the excretory ducts and the adventitia were determined. Conclusion: Sheep castration promoted changes in the biometric measures of the glands, which were lower in castrated animals. The morphological and biometric characteristics of the vesicular and bulbourethral glands in sheep were determined.

Anthelmintic efficacy of an oral formulation of Aurixazol against gastrointestinal nematodes of naturally and experimentally infected sheep

As a result of the need to develop new active principles for the control of endoparasites in ruminants, the present in vivo study evaluated a formulation containing 24% Aurixazol (48 mg/kg), a parasiticide molecule based on disophenolate of levamisole. Two experiments were conducted: one evaluating the anthelmintic efficacy of 24% Aurixazol (48 mg/kg) against gastrointestinal nematodes in naturally infected sheep, compared to an association of ivermectin (0.2 mg/kg) + albendazole (5.0 mg/kg) + levamisole (7.5 mg/kg) (IAL), and a second one which evaluated the persistent efficacy of the same formulation against immature stages (L4) and adults of Haemonchus contortus in experimentally infected animals. In experiment I, against H. contortus, the formulation of Aurixazol and the IAL association reached efficacies (arithmetic means) of 99.32% and 96.11%, respectively. For Trichostrongylus colubriformis, the efficacy values were 88.92% and 98.08% for Aurixazol and the IAL association, respectively. Both formulations were totally effective against Oesophagostomum columbianum (100%). The results of the statistical analysis demonstrated that the mean parasitic burden of treated animals was significantly different (P ≤ 0.05) compared to the average number of helminths diagnosed in animals from the control group for H. contortus, T. colubriformis and O. columbianum. Comparing only the treated groups, it was possible to verify that the average number of H. contortus recovered from animals treated with Aurixazol was different (P ≤ 0.05) when compared to the mean amount recovered from sheep treated with the IAL association. When evaluating the prevention of H. contortus infection in experiment II, Aurixazol did not present preventive efficacy. Up until 21 days after treatment the groups treated with Aurixazol contained less adults and L4 of H. contortus (P ≤ 0.05) when compared to the non-medicated control group. However, future studies will be necessary to assess the effectiveness of Aurixazol against nematode strains resistant to levamisole and disophenol, but the efficacy results described in this study allow to state that Aurixazol can, associated with other measures, become an important tool in the control of sheep nematodes. © 2013.

In vitro and in vivo evaluation of the activity of pineapple (Ananas comosus) on Haemonchus contortus in Santa Inês sheep

The development of resistance to anthelmintics has prompted research into alternative methods of controlling intestinal nematodes in ruminants. This study aimed to assess the activity of Ananas comosus on Haemonchus contortus in Santa Inês sheep. The aqueous extract of pineapple skin (AEPS), bromelain from pineapple stems (B4882) and residue from pineapple processing was evaluated in in vitro and in vivo tests. The enzymatic activity of substances was analyzed by the azocasein method. The egg hatch test (EHT) and larval development test (LDT) were performed using the Embrapa2010 isolate of H. contortus. In the in vivo test, 36 sheep artificially infected with H. contortus were divided into six groups: G1: 2g/kg BW of the aqueous extract administered for three days; G2: 2g/kg BW of the industrial pineapple residue for 60 days; G3: 180mg/animal of bromelain in a single dose; G4: negative control I; G5: positive control (levamisole phosphate); and G6: negative control II. The eggs per gram (EPG) in the feces were counted till 28 days after treatment. LC50 and LC90 were obtained by the probit procedure, while the in vivo test results were analyzed by GLM. The aqueous extract in the in vitro and in vivo test, the bromelain and industrial residue presented 0.102, 0.157, 1.864 and 0.048 enzyme units/mL, respectively. In the egg hatch test, the LC50 and LC90 were respectively 31 and 81mg/mL for the aqueous extract and 0.50 and 2mg/mL for bromelain. In the larval development test, the LC50 and LC90 were respectively 1.7 and 7.3mg/mL for the aqueous extract and 0.019 and 0.086mg/mL for bromelain. In the in vivo test, the general efficacies of the treatments in relation to the negative control were 22.6%, 42.2%, 3.65% and 89% for the aqueous extract, industrial pineapple residue, bromelain and positive control respectively. The transformed EPG values were 3.19±0.59, 3.32±0.25, 2.85±0.66, 3.44±0.50, 2.28±0.93 and 2.75±0.94 for the aqueous extract, industrial residue, bromelain, negative control I, positive control and negative control II respectively. The results for all the treated groups differed significantly (p<0.05) from the positive control, and although the residue presented efficacy of 42.2%, there was no statistical difference (p>0.05) in relation to the negative control. Therefore, both the aqueous extract and bromelain were effective in vitro, but showed reduced anthelmintic efficacy in vivo. For the pineapple residue, the 42.2% in vivo efficacy in reducing the EPG and the possibility of reducing environmental contamination through reuse of industrial residue indicate it can also be useful for control of this parasite. © 2013 Elsevier B.V.

Nutrient intake and digestibility of the lipid residue of biodiesel from palm oil in sheep

ABSTRACT: The objective of this study was to evaluate the effect of inclusion of lipid residue of biodiesel originated in the processing of palm oil (Elaeis guineensis) in the diet on the digestibility of feedlot lambs. Twenty-five crossbred male castrated lambs, weighing 20±1.61 kg, were distributed in randomized blocks with five treatments and five replications. The experimental period lasted 22 days; 15 for diet adaptation, 2 for the adaptation to the indicator LIPE (lignin from Eucalyptus grandis isolated, purified and enriched, UFMG, Minas Gerais) and 5 for fecal sampling. Diets were formulated with 64% concentrate based on corn and soybean meal, 31% Massai grass (Panicum maximum cv. Massai) hay and 5% lipid supplementation from increasing levels of substitution of 0, 25, 50, 75 and 100% of palm oil for biodiesel oil from palm residue. The lambs were offered two meals a day, at 7h00 and 16h00. There was linear effect of inclusion of the residue from palm oil biodiesel on dry matter intake. There was no change in digestibility of nutrients except for ether extract. The use of biodiesel from palm oil residue up to 100% replacement for the lipid supplementation of sheep positively influences the consumption without altering the digestibility of nutrients.

Effect of alternate and simultaneous grazing on endoparasite infection in sheep and cattle

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Production of sheep grazing on nitrogen-fertilized tifton 85 grass in continuous stocking

Biomass and forage volume density and the performance and stocking rate of sheep on pastures with nitrogen-fertilized Tifton 85 and subjected to a continuous stocking system were evaluated. Four doses of nitrogen (0, 100, 200, and 400 kg ha-1 year), arranged in an experimental design with randomized blocks and four replications, were analyzed. Sixteen paddocks and Santa Inês sheep were used as test animals, coupled to crossbreed Santa Inês sheep as regulating animal stocking. Nitrogen-fertilized Tifton 85 pastures increased the amount of forage biomass and volume density which affected stocking rate and weight gain of sheep in continuous grazing. When pastures with Tifton 85 were administered in variable load continuous stocking, with grass kept at 15 cm, nitrogen fertilization up to 400 kg ha-1 year is recommended.

Tropical tanniniferous legumes used as an option to mitigate sheep enteric methane emission

This study presents the first results from Brazil using SF6 tracer technique adapted from cattle to evaluate the capability of condensed tannin (CT) present in three tropical legume forages, Leucaena leucocephala (LEU), Styzolobium aterrimum (STA), and Mimosa caesalpiniaefolia Benth (MIM) to reduce enteric CH4 production in Santa Ins sheep. Twelve male lambs [27.88 +/- 2.85 kg body weight (BW)] were allocated in individual metabolic cages for 20-day adaptation followed by 6 days for measuring dry matter intake (DMI) and CH4 emission. All lambs received water, mineral supplement, and Cynodon dactylon v. coast-cross hay ad libitum. The treatments consisted of soybean meal (710 g/kg) and ground corn (290 g/kg) [control (CON)]; soybean meal (150 g/kg), ground corn (30 g/kg), and Leucaena hay (820 g/kg) (LEU); soybean meal (160 g/kg), ground corn (150 g/kg), and Mucuna hay (690 g/kg) (STA); and soybean meal (280 g/kg), ground corn (190 g/kg), and Mimosa hay (530 g/kg) (MIM); all calculated to provide 40 g/kg CT (except for CON). DMI (in grams of DMI per kilogram BW per day) was lower for LEU (22.0) than CON (29.3), STA (31.2), and MIM (31.6). The LEU group showed emission of 7.8 g CH4/day, significantly lower than CON (10.5 g CH4/day), STA (10.4 g CH4/day), and MIM (11.3 g CH4/day). However, when the CH4 emission per DMI was considered, there were no significant differences among treatments (0.37, 0.36, 0.33, and 0.35 g CH4/g DMI/kg BW/day, respectively, for CON, LEU, STA, and MIM). The sheep receiving STA had shown a tendency (p = 0.15) to reduce methane emission when compared to the CON group. Therefore, it is suggested that tropical tanniniferous legumes may have potential to reduce CH4 emission in sheep, but more research is warranted to confirm these results.

High occurrence of Cryptosporidium ubiquitum and Giardia duodenalis genotype E in sheep from Brazil

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Leptospira spp. infection in sheep herds in southeast Brazil

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)