A nontoxic derivative of Bothrops jararaca venom suitable to generate antibodies against the native venom.

Bothrops jararaca venom was toxoided by stepwise iodination with cold iodine, and doses up to 30 LD50 were non-lethal by i.p. route (BICALHO et al., 1990). Groups of mice injected chronically with the native, or the iodinated venom, have been subjected to histological examination. In the native group, in the spleen, around the white pulp, an acellular, amorphous eosinophilic substance, metachromatic to Methyl Violet, PAS positive, and dichroic to Congo Red under polarized light, was present. Strong congestion in the liver, kidneys and lungs was found. The salivary glands were replenished with an amorphous substance in the serosal acini. The groups injected with the iodinated venom only show discrete alterations, more akin to the control group. The anavenin was immunogenic. Antibody generation in mice and rabbits was detected by ELISA. In mice, active protection against challenge with native venom was presented. The iodinated venom generated a rabbit antiserum with strong lines in gel immunoprecipitation against the lethal venom. A minimum neutralization titer of 2.3 mg ml-1 against the native venom was attained in the first cycle (28 days) of immunization. After 3 cycles (100 days), the protection rose to 5.1 mg ml-1.     

The expression of plasmid mediated afimbrial adhesin genes in an avian septicemic Escherichia coli strain

An Escherichia coli strain (SEPT13) isolated from the liver of a hen presenting clinical signs of septicaemia had a LD₅₀ of 4.0 × 10⁵ CFU/ml in one-day-old chickens, expressed Ia, Ib, E1, E3, K and B colicins and aerobactin. The strain was ampicillin and streptomycin resistant, and found to have fimA, csgA and tsh DNA related sequences; it could adhere to and invade HEp-2 and tracheal epithelial cells, expressed fimbriae (observed by electron microscopy), and had five plasmids of 2.7, 4.7, 43, 56, and 88 MDa. Transposon mutagenesis of strain SEPT13, with transposon TnphoA, resulted in a mutant strain named ST16 that had a LD₅₀ of 1.2 × 10¹² CFU/ml. All other biological characteristics of strain ST16 were the same as those detected for strain SEPT13 except for the migration of an 88 MDa plasmid to the 93 MDa position indicating the insertion of the transposon into the 88 MDa plasmid. The 93 MDa plasmid of strain ST16 was transferred, by electroporation assay, to non-pathogenic receptor strains (E. coli strains K12 MS101 and HB101), resulting in transformant strains A and B, respectively. These strains exhibited adhesion properties to in vitro cultivated HEp-2 cells but did not have the capacity for invasion. The adherence occurred despite the absence of fimbriae; this finding suggests that the 88 MDa plasmid has afimbrial adhesin genes.     

Causes of bruising in carcasses of beef cattle during farm,transport, and slaughterhouse handling in Brazil

This study aimed to assess preslaughter factors responsible for the occurrence of bruises to front, rib, loin, hip, and round sites of cattle carcasses. Data from the slaughter of 148 loads of cattle were assessed, totaling 4,611 carcasses. Evaluated bruising factor variables were animal gender, journey distance (km), vehicle type (m²), journey time (hours), load density (kg/m²), handling and facilities conditions (optimal, acceptable, and unacceptable), and unloading waiting time (min). Gender had the strongest influence on carcass bruising, being the first variable to enter in the majority of models related to the carcass sites. Female carcasses displayed higher bruising in all sites. Densities above 401 kg/m² caused more bruising. With the exception of the loin, unacceptable, and acceptable conditions assigned to farm facilities and handling resulted in higher average bruising. Articulated vehicles with a greater load capacity were more likely to cause bruising. The effects of variables causing bruises act differently in the various carcass sites. In this sense, the results of this study suggested possible management practices for specific problems during the preslaughter period to minimize losses by bruising to different carcass sites.     

Effects of Growth Hormone on In Situ Culture of Bovine Preantral Follicles are Dose Dependent

The objective of this study was to evaluate different concentrations of growth hormone (GH) on the development of bovine preantral follicles cultured included in the ovarian tissue (in situ) on the rates of morphologically normal, viable, primordial and developing follicles, as well as the oocyte and follicle diameter and ultrastructural analysis. Ovarian fragments collected from cows with no cross‐breeds defined were cultured in situ for 1 and 7 days in minimal essential medium (α‐MEM+) supplemented with different concentrations of recombinant human GH (0, 10, 25, 50 ng/ml). The ovarian fragments non‐cultured (control) and cultured were processed for classic histology, mechanical isolation and electron transmission microscopy (MET). The parameters underwent anova (Tukey′s and Dunnett′s tests) and chi‐square test (χ²). After 7 days of culture, the treatment with 50 ng/ml GH showed no differences with fresh control (p > 0.05) and had greater effectiveness than in the 0, 10 and 25 ng/ml GH concentrations of the morphologically normal follicles. Regarding the primordial follicles, a reduction was observed in the 50 ng/ml GH concentration concomitant with the significant increase in developing follicles, differing from both the fresh control and the other GH concentrations tested. In addition, 50 ng/ml GH showed a larger follicle and oocyte diameter when compared to the other treatments cultured. Similar structures were ultrastructurally observed in the control group, 50 ng/ml GH. Follicles cultured in 10 ng/ml GH showed nuclear invagination, vacuoles and lesioned basal membrane. Hence, it is concluded that 50 ng/ml GH is the most effective concentration for the development of preantral follicles cultured in situ.     

Serum Levels of Cardiac Markers NT‐proANP and NT‐proBNP in Brachycephalic bitches at Different Gestational Stages

The aim of this study was to determine serum levels of natriuretic peptide precursors (NT‐proANP and NT‐proBNP) during pregnancy in brachycephalic bitches. Fifteen healthy multiparous bitches were selected for this prospective study. Serum levels of NT‐proANP and NT‐proBNP were measured during anoestrous and at 14, 35, 42, 49 and 56 days (2nd, 5th, 6th, 7th and 8th weeks) of pregnancy. Fourteen animals had normal gestations, and one bitch developed single foetus syndrome. The natriuretic peptide levels of this animal were not included in this study; however, it is important to report that its NT‐proANP levels were four times greater than those of normal patients. There was no significant difference (p = 0.072) in NT‐proBNP levels between anoestrous (0.20 ± 0.10 ng/ml) and the different pregnancy weeks (0.27 ± 0.12 ng/ml). There was a positive correlation (p < 0.0001) between NT‐proANP and gestational age, and the levels of this marker increased significantly (p < 0.0001) during the 6th (0.26 ± 0.06 ng/ml), 7th (0.28 ± 0.04 ng/ml) and 8th weeks (0.29 ± 0.05 ng/ml) when compared to anoestrous (0.18 ± 0.02 ng/ml). NT‐proANP serum levels are correlated with gestational development and may be indicative of cardiovascular adaptation in canine brachycephalic pregnancy.     

In vitro evaluation of cryopreserved bovine sperm and its relation to field fertility in fixed‐time artificial insemination

This study aimed to assess characteristics of bovine cryopreserved sperm and evaluate its relation to field fertility in fixed‐time artificial insemination (FTAI). Semen samples of 16 bulls were used to inseminate 811 Nellore cows, and four of these bulls were also used to inseminate 101 Nellore heifers. Samples of the same ejaculate used for FTAI from each bull were analysed in the laboratory after thawing. Sperm motility and vigour were subjectively assessed by light microscope, and integrity of the plasma and acrosome membranes, and H₂O₂ production were evaluated by flow cytometer. Relation among sperm characteristics and pregnancy rate of cows and heifers were evaluated by univariate and multivariate logistic regression. Subjective sperm motility and vigour did not affect the probability of pregnancy in cows or heifers. In univariate analysis for pregnancy in cows, sperm traits related to acrosome injury positively affected probability of pregnancy mainly when associated with plasma membrane integrity; H₂O₂ production seems to be less important than plasma membrane integrity in affecting probability of pregnancy. In multivariate analysis, sperm traits related to injured acrosome positively affected probability of cow and heifer pregnancies while intact acrosome was negatively related to cow pregnancy. Intact plasma membrane and high H₂O₂ production were positively related to cow pregnancy but negatively related to heifer pregnancy. Results suggest that a capacitation‐like status of the acrosome may benefit probability of pregnancy in cows.     

Variance component estimations for race performance of thoroughbred horses in Brazil by random regression model

A total of 306,698 racing performance data recorded between 1992 and 2002 from 25495 Brazilian Thoroughbred horses was analyzed to estimate the variance components using the Random Regression Model (RRM) compared to the Classical Repeatability Animal Model (CRAM). The performance was evaluated using the race time (in seconds) to run distances of 1000, 1100, 1200, 1300, 1400, 1500 or 1600 meters. The pedigree of each horse was extended as far as possible, with a total of 36,659 animals. The simple correlation between Breeding Value (BV) for the race time estimated by both procedures was high (r = 0.963) when the data were expressed at the mean level of all distances. However, with the RRM it was possible to estimate the genetic parameters and the BV of all animals at the trajectory of each one of the seven distances. Also, results clearly showed that a single BV estimated by CRAM, is neither an adequate nor a sufficient indicator for the selection of the best horses throughout the trajectory of distance performance. We concluded that the RRM procedure is highly recommended for the evaluation of racehorse performance.     

The importance of vertical transmission of Neospora sp. in naturally infected horses

Neospora spp. is a intracellular protozoan phylogenetically closely related to Toxoplasma gondii and Sarcocystis neurona, and it can infect horses leading to the development of reproductive or neurological diseases. We determined the presence of antibodies to Neospora sp. in mares at their parturition time and determine the frequency of vertical transmission in healthy foals to verify the importance of transplacental transmission. The samples were analyzed by indirect immunofluorescence antibody test, showing that seroprevalence in mares is higher than in foals and seropositive mares are likely to transmit the neosporosis to their offspring. This shows that endogenous challenge occurs in horses, and it suggests that this protozoan can be disseminated by means of transplacental transmission in horse species.     

Effect of rutin and chloroquine on White Leghorn chickens infected with <Emphasis Type="Italic">Plasmodium (Bennettinia) juxtanucleare</Emphasis>

Plasmodium juxtanucleare is one of the agents that cause avian malaria in chickens and little is known about the treatment of the infection. The aim of this study was to compare the antimalarial activities of rutin and chloroquine in chickens infected with P. juxtanucleare after immunossuppression with corticosteroid. Antimalarial activity was verified in 33 fowls by evaluation of parasitemia levels in Giemsa-stained blood smears taken to 30 days. Rutin did not exert a reduction in parasitemia from P. juxtanucleare when compared to chloroquine treatment that kept the parasites at a low level, demonstrating its antimalarial effect. During the course of the infection, the trophozoite stage predominated (80%), followed by schizont (17%) and gametocyte (3%). Maximum parasitemia levels were recorded on day 12 in control and rutin groups. There were no significant differences in the hematocrit values, weight body or the temperature of the fowls among the groups evaluated (p > 0.05).     

Platelet-Rich Plasma in the Treatment of Induced Tendinopathy in Horses: Histologic Evaluation

This study was carried out to evaluate the effect of platelet-rich plasma (PRP) on the treatment of tendinopathy induced in the superficial digital flexor tendon (TFDS) of horses, by using histologic evaluation. Six healthy crossbred geldings aged 8 to 15 years (12 ± 3) were used. The TFDS tendinopathy was provoked in both forelimbs, by intratendinous administration of 2.5 mg collagenase (2.5 mg/mL), and this procedure was considered as the beginning of the experimental phase. At 12 days after induction of the tendinopathy, the animals were subjected to the following treatments: (1) in the lesion caused in the right superficial digital flexor tendon (PRP-treated group), 2.5 mL PRP activated with calcium chloride at 0.0125 mol/L at concentrations from 320,000 to 500,000 platelets/μL, were injected; (2) in the tendinopathy of the left SDFT (control group), 2.5 mL 0.9% saline solution was administrated. Thirty-six days after the treatments, a biopsy of the injured area was performed for histologic evaluation. In both groups, the histologic analysis showed an increase in the fibroblastic density, as well as the presence of neovascularization, lymphocytes, and plasmocytes infiltrate and tissue organization at variable intensity. In the PRP-treated group, the SDFT was more organized, with the collagen fibers and fibroblasts being better arranged on the tendon matrix. The numbers of the fibroblasts and blood vessels did not differ between the groups. Histologic evaluation 36 days after tendinopathy showed that injuries under a single PRP treatment present a more uniform and organized tissue repair when compared with the control group.