Evaluation of an enzyme-linked immunosorbent assay for diagnosis of trichinellosis in swine.

Experimental and field trials were conducted to evaluate an ELISA for its ability to detect Trichinella-infected domestic swine and to compare ELISA results with muscle-digestion test results. The ELISA used was a commercial double-antibody kit, containing an excretory-secretory antigen, and was evaluated principally for epidemiologic use. Experimentally induced infection in swine (4 groups of 3 pigs each; inoculated with 0, 50, 500 or 5,000 larvae) was detected as early as postinoculation week 4, with seroconversion of all inoculated swine by postinoculation week 8. The rate of seroconversion appeared to be affected by initial larval dose, time after inoculation, and immunocompetence of the individual host. Determination of antibody kinetics generally revealed rapidly increasing antibody titer, followed by its steady decrease in most pigs. Once seropositive, however, all pigs remained seropositive for the duration of the 10-week study. Presence of muscle larvae was confirmed in all infected pigs at termination of the study. We recognize that the experimental conditions may not be truly representative of those under which natural infection develops in pigs; however, the ELISA detected an infected pig with muscle larval density of 0.87 larvae/g of tissue. Results of a field trial (n = 310) indicated no muscle digestion test-positive pigs (35 g of diaphragm muscle digested/pig), but 3 samples tested positive by ELISA for specificity of 99.0%.     

In Vivo Embryo Recovery Rate by Laparoscopic Technique from Rabbit Does Selected for Growth Rate

Rabbit does from R line selected for growth rate present a low reproductive performance and this study aimed to evaluate both the recovery efficacy and viability of recovered embryos after vitrification and the reproductive performance of donor does subjected to in vivo recovery. Does were divided into three groups: 28 does without in vivo recovery (control), 25 does in which in vivo recovery was started in the nulliparous state (group 1) and 30 does with at least one litter before in vivo recovery (group 2). Does were superovulated with a single subcutaneous injection of 50 IU of equine chorionic gonadotropin (eCG) per female, and were then artificially inseminated 60 h later and immediately administered an intravenous dose of 75 IU of human chorionic gonadotropin (hCG) per female. Does from group 1 and 2 were recovered in vivo 76-80 h post-insemination by repeated laparoscopies at one to four times and permitted one or two parturitions between recoveries [in vivo (IV) recovery]. At the end of the experiment, about 16 does of all groups were recovered post-mortem (PM recovery). All normal embryos were vitrified, devitrified and then cultivated in vitro to evaluate the viability after thawing. A significant increase in the ovulation rate was found in does recovered PM than in those recovered IV in the nulliparous state. However, no significant differences were observed in the recovery rate, the donor rate, the number of normal embryos recovered with at least one normal embryo per doe and the viability after thawing between the PM and IV groups. A significant decrease in the fertility rate, total born, live born and weaned kids was found for does from group 1 in comparison with does from group 2. Results support the use of repeated laparoscopy to increase the number of recovered embryos per donor doe especially in such R line does, if they are permitted to produce at least one litter before the beginning of in vivo recovery.     

Use of Powdered Egg Yolk vs Fresh Egg Yolk for the Cryopreservation of Ovine Semen

Egg yolk is a common additive to sperm cryopreservation diluents. Because of its animal origin, however, it also represents a potential risk of microbiological contamination in the diluent. This potential contamination can be avoided by using powdered egg yolk, instead of fresh egg yolk, as it is pasteurized. This study was conducted to determine ram sperm cryosurvival was affected by the type of egg yolk used (powdered egg yolk or fresh egg yolk) and by yolk concentration (10, 15 or 20%) in the diluent. Microbiological analyses were also performed to quantify the microbiological contamination in the diluents containing the two types of egg yolk. Sperm cryosurvival was determined by motility and morphology analyses after thawing. Motility parameters were assessed using a computer-assisted sperm analysis (CASA) system, and the percentage of sperm with a normal apical ridge was evaluated using a differential interference contrast microscope. No significant differences were observed between diluents in the percentage of sperm with normal apical ridge. However, higher percentages of total motile cells were observed for samples containing powdered egg yolk (69%) compared to samples containing fresh egg yolk (60%). However, sperm in diluents containing fresh egg yolk, exhibited higher values for average-path velocity, straight-line velocity and beat cross frequency and lower values for amplitude of lateral head displacement (p <0.05), compared to cells in diluents containing powdered egg yolk. Microbiological contamination was similar (<200 CFU/ml) in both diluents, and no bacterial growth was observed in either, when antibiotics were added. Therefore, powdered egg yolk can be effective used in diluents for the freezing of ram semen. However, the in vivo fertility of sperm frozen in diluents containing powdered egg yolk should be tested, as some motility parameters were different for sperm treated with powdered egg yolk compared to fresh egg yolk.     

Death of a neonatal lamb due to Clostridium perfringens type B in New Zealand

ABSTRACT

Case history and clinical findings: A flock of 20 sheep was kept within three paddocks on a single property. None of the animals in the flock had been vaccinated against any disease for at least three years. Abdominal bloating and haemorrhagic diarrhoea were observed in Lamb 1 at 24 hours-of-age. The lamb subsequently died within an hour of the onset of clinical signs. Lamb 2 was 3-days-old when observed to be recumbent with opisthotonus. The lamb was treated with dextrose, vitamins B1 and B12, and penicillin G, but died 4 hours later.

Pathological findings: Examination of Lamb 1 revealed markedly increased gas within the peritoneum and within dilated loops of intestine. The intestines were dark red and contained large quantities of haemorrhagic fluid. Histology of the intestines revealed peracute mucosal necrosis with minimal accompanying inflammation. The intestinal lumen contained cell debris, haemorrhage, and myriad large Gram-positive bacilli. The intestines of Lamb 2 did not appear bloated or reddened. However, multiple fibrin clots were visible within the pericardial sac. Histopathological examination revealed small foci of necrosis within the mucosa of the distal intestine. The necrotic foci were often associated with large numbers of large Gram-positive bacilli.

Immunohistochemsitry and molecular biology: Intestinal samples from Lamb 1 were processed for Clostridium perfringens immunohistochemistry, which revealed large numbers of intralesional, positively immunostained rods. Fragments corresponding to the expected sizes for genes encoding alpha, beta, and epsilon C. perfringens typing toxins were amplified by PCR from DNA extracted from formalin-fixed sections of intestine.

Diagnosis: Lamb dysentery due to C. perfringens type B.

Clinical relevance: C. perfringens bacteria have a worldwide distribution, but disease due to C. perfringens type B has only been diagnosed in a small number of countries and has never been reported in New Zealand or Australia. C. perfringens type B produce both beta toxin and epsilon toxins, therefore both haemorrhagic enteritis and systemic vascular damage can develop. As many animals are exposed to C. perfringens without developing disease, there must be additional unknown factors that resulted in disease in these particular sheep. Vaccines that specifically protect against C. perfringens type B are available and may be recommended for use in smaller non-commercial flocks, as in the present case.     

Pre-partum monensin supplementation improves body reserves at calving and milk yield in Holstein cows dried-off with low body condition score

The objective of this study was to determine the effect of a monensin controlled-release capsule administered intraruminally at drying-off on body condition score (BCS) at calving, milk yield, fertility and concentration of energy-related blood metabolites in Holstein cows dried-off with low BCS (< or = 3.0, scale 1 to 5 with a 0.25 point of increment). Between July and August, 2001, 220 cows from parity 2 or more and dried-off 50-70 days before expected parturition, with a BCS < or = 3.0 were randomly assigned to either a treatment group (n=110; oral capsule of monensin releasing 335 mg/day for 95 days) or a control group (no capsule, n=110). At assignment, on day 21 before expected parturition, at calving, and at 7, 14, and 21 days in milk a blood sample was obtained from a random sub sample of 10 cows per group. Effects of monensin on serum NEFA, BHBA and glucose were measured. Milk yield, milk fat and protein content (%) at DHIA test days during the entire lactation, 305 ME milk production and reproductive responses were compared. Monensin significantly improved BCS at calving, increased milk yield at test days 4 and 8, decreased the percentage of milk protein, did not change the percentage of milk fat, and decreased NEFA and BHBA during the post-partum period.     

Retrospective assessment of tolerability and efficacy of zoledronate in the palliative treatment of cancer-bearing dogs

Zoledronate is a bisphosphonate frequently used for the treatment of hypercalcaemia of malignancy and tumour-associated bone pain in dogs, however, there is a paucity of information regarding its use in veterinary medicine. The aim of this retrospective study was to report the tolerability of zoledronate in the palliative treatment of cancer-bearing dogs and secondarily to to assess the efficacy of zoledronate for the treatment of hypercalcaemia of malignancy. Thirty-seven dogs (22 with tumour-associated bone pain and 15 with hypercalcaemia of malignancy) that received 114 zoledronate infusions were included. Tolerability was assessed by the absence of post-zoledronate hypocalcaemia or other adverse events as defined by Veterinary Cooperative Oncology Group-Common Terminology Criteria for Adverse Events criteria. Efficacy was assessed by comparison of available ionized calcium levels before and after zoledronate administration in hypercalcaemic dogs. In 79% of zoledronate infusions, no adverse events were reported. The majority of adverse events which occurred in the other 21% of infusions could be attributed to concurrent chemotherapy or the underlying neoplastic disease. There was a small but significant increase in creatinine following treatment with zoledronate, however, none of the dogs developed clinically significant renal disease. In eight hypercalcaemic dogs with available ionized calcium following zoledronate administration, ionized calcium decreased rapidly within 7 days following treatment with zoledronate. Zoledronate is well-tolerated with few recorded adverse events, however, monitoring of serum creatinine is advised. Zoledronate seems to be effective in the treatment of hypercalcaemia of malignancy.     

Chemical characteristics and relative bioavailability of supplemental organic copper sources for poultry

Five commercially available organic Cu products and reagent-grade CuSO4 x 5H2O (Cu Sulf) were evaluated by polarographic analysis and solubility in 0.1 M K2HPO4-KH2PO4 buffer (pH 5), 0.2 M HCl-KCl buffer (pH 2), or deionized water. Fractions from these solubility tests were evaluated by gel filtration chromatography for structural integrity. The organic sources were Cu lysine complex (Cu Lys), Cu amino acid chelate (Cu AA), Cu proteinate A (Cu ProA), Cu proteinate B (Cu ProB), and Cu proteinate C (Cu ProC). Separation of peaks in the chromatograms for the soluble Cu fraction from deionized water indicated that 77, 31, 69, 94, and 16% of the Cu remained chelated for the above sources, respectively. Two experiments were conducted to estimate the relative bioavailability of Cu from the organic Cu supplements for chicks when added at high dietary concentrations to practical corn-soybean meal diets. Liver Cu concentration increased (P < 0.0001) as dietary Cu increased in both experiments. When Cu Sulf was assigned a value of 100% as the standard, linear regression slope ratios of log10 liver Cu concentration regressed on added dietary Cu concentration gave estimated relative bioavailability values of 124 +/- 5.1, 122 +/- 5.3, and 111 +/- 6.0 for Cu Lys, Cu AA, and Cu ProC, respectively, in Exp. 1. The bioavailability estimates for Cu Lys and Cu AA were greater (P < 0.05) than that for Cu Sulf. Values in Exp. 2 were 111 +/- 7.6, 109 +/- 8.4, and 105 +/- 7.5 for Cu Lys, Cu ProA, and Cu ProB, respectively, and all sources were similar in value for chicks. Solubility of Cu in pH 2 buffer provided the best prediction of bioavailability (r2 = 0.924). Other indicators of chelation integrity and solubility had little value as predictors of bioavailability (r2 < or = 0.445).     

Chemical characteristics and relative bioavailability of supplemental organic zinc sources for poultry and ruminants

Eight commercially available organic Zn products and reagent-grade ZnSO4 x 7H2O (Zn Sulf) were evaluated by polarographic analysis, and solubility in .1 M K2HPO4-KH2PO4 buffer (pH 5), .2 M HCl-KCl buffer (pH 2), and deionized water. Fractions from these solubility tests were evaluated by gel filtration chromatography for structural integrity. Degree of chelation was generally positively related to chelation effectiveness determined by polarography. The organic sources were Zn methionine complex A (Zn MetA), Zn methionine complex B (Zn MetB), Zn polysaccharide complex (Zn Poly), Zn lysine complex (Zn Lys), Zn amino acid chelate (Zn AA), Zn proteinate A (Zn ProA), Zn proteinate B (Zn ProB), and Zn proteinate C (Zn ProC). Three experiments were conducted to estimate the relative bioavailability of Zn from the organic Zn supplements for chicks and lambs when added at high dietary levels to practical diets. Bone Zn concentration increased (P < .001) as dietary Zn increased in both experiments. When Zn Sulf was assigned a value of 100% as the standard, multiple linear regression slope ratios of bone Zn from chicks fed 3 wk regressed on dietary Zn intake gave estimated relative bioavailability values of 83 +/- 14.6 and 139 +/- 16.9 for Zn AA and Zn ProA, respectively, in Exp. 1 and 94 +/- 11.6, 99 +/- 8.8, and 108 +/- 11.4 for Zn Poly, Zn ProB, and Zn ProC, respectively, in Exp. 2. In Exp. 3, 42 lambs were fed diets containing Zn Sulf, Zn ProA, Zn AA, or Zn MetB for 21 d. Based on multiple linear regression slope ratios of liver, kidney, and pancreas Zn and liver metallothionein concentrations on added dietary Zn, bioavailability estimates relative to 100% for Zn Sulf were 130, 110, and 113 for Zn ProA, Zn AA, and Zn MetB, respectively. Except for Zn ProA, which was greater, the organic Zn supplements had bioavailability values similar to that of Zn Sulf for chicks and lambs. Bioavailability of organic Zn products was inversely related to solubility of Zn in pH 5 buffer in chicks (r2 = .91) and pH 2 buffer in lambs (r2 = .91), but not to an estimate of degree of chelation.     

Bluetongue virus serotypes 1 and 3 infection in Poll Dorset sheep

Objective To study the clinical signs following bluetongue virus serotypes 1 and 3 infection in Poll Dorset sheep.
Design A clinical and pathological study.
Procedure Twenty Poll Dorset sheep were inoculated with bluetongue virus serotypes 1 or 3, each inoculum having a different passage history. The sheep were examined daily and their clinical appearance and rectal temperatures recorded. Heparinised and non-heparinised blood samples were taken at intervals for virological and serological study. Gross pathological findings were recorded for several sheep at necropsy and tissue samples were collected from three sheep for virological studies.
Results All inoculated sheep developed clinical disease. The clinical signs and gross pathological changes varied considerably but were consistent with damage to the vascular endothelial system. There was a decline in the titres of infectious bluetongue virus and of antigen in tissues collected between 7 and 12 days after infection.
Conclusions The severity of disease was related to the speed of onset and duration of pyrexia and not the development or titre of viraemia. Generally, those animals with sensitive mouths, depression, coronitis, recumbency and reluctance to move were the most debilitated. Whole blood was the most reliable source of infectious virus from acutely and chronically infected and convalescent animals. However, tissue samples particularly spleen, collected from dead or killed animals suffering from either peracute or acute forms of disease were most appropriate for the rapid confirmation of a clinical diagnosis.     

Unguided bronchoalveolar lavage techniques and residual effects in dogs

Objectives To investigate the use of unguided bronchoalveolar lavage techniques in dogs without fibreoptic bronchoscopy, using an adapted single vascular catheter and a double-lumen catheter made from two single vascular catheters.
Animals Sixty-nine dogs were examined with the single-catheter technique and 110 dogs with the double-catheter technique.
Design A prospective study.
Procedure Sixty-nine and 220 samples, collected with the single catheter and the double catheter respectively, were examined cytologically Lungs of 69 dogs were examined grossly and histologically. Radiographic examination was performed on 11 dogs.
Results The double-catheter technique produced samples with significantly higher cellularity (P < 0.01) and fewer red blood cells (P < 0.01) than the single-catheter technique. Repeat samples collected with a double catheter were not significantly different (P > 0.01) in any value. A reference range for nucleated cell counts of 62 to 1210 – 10⁶/L was calculated from 57 clinically and histologically normal dogs. The major residual effects of the technique were localised pulmonary oedema, and alveolar distension with collapse and congestion of distant parenchyma. Thoracic radiographs revealed increased lung opacity for at least up to 7 h after the procedure.
Conclusions The cellularity of the bronchoalveolar lavage fluid obtained was adequate and sufficient fluid was retrieved when the single catheter was located in a proper position. However, the double catheter obtained better samples more quickly and easily, with less damage to the respiratory tract.