Enzyme Immunoassays for the Determination of Ovine LH and FSH

The development of competitive enzyme immunoassays for ovine plasma LH (oLH) and FSH (oFSH) is described. Standards and plasma samples were preincubated with diluted antiserum to oLH or oFSH and the reacted solution (100 μl per well) was transferred to plates previously coated with oLH or oFSH, respectively. The second antibody used was anti‐rabbit IgG horseradish peroxidase. The measuring range was 0.39–50 ng/ml for each hormone and the 50% relative binding sensitivity was 9 ng/ml for oLH. The respective value for oFSH was 3.5 or 34 ng/ml with different hormone and antibody preparations used for the assay. The enzyme immunoassays were used to determine oLH and oFSH levels in plasma from ewes of two breeds during the oestrous cycle. The assays detected the first FSH surge coincident with the LH surge, the second FSH surge about 24 h later and the periodic fluctuations of FSH concentrations during the luteal phase of the oestrous cycle. These enzyme immunoassays are an efficient and economic alternative to the established radioimmunoassays (RIA) for oLH and oFSH.     

Chronic vascular catheterisation of the koala and the rabbit

SUMMARY: Vascular access ports were surgically placed, first in rabbits maintained under laboratory conditions, and then in koalas maintained in a wildlife park. The ports remained patent for 9 months in rabbits and for up to 13 months in the koalas and were removed successfully. They allowed collection of blood samples without assistance or disturbance in koalas, and without stress as reflected by plasma cortisol concentration. The use of retention rings on the cannula tubing is recommended.     

Alcohol diluent provides the optimal formulation for calcium chloride non-surgical sterilization in dogs

Background

Surgical castration is widely used to sterilize male dogs, but has significant impacts on time to perform the operation, recovery of the animals as well as cost, which can limit population control programs. Previous research has shown intratesticular injection of calcium chloride dihydrate (CaCl₂) in saline to be a promising alternative to surgery. However, long-term azoospermia was not maintained at dosages low enough to avoid side effects. In the search for an optimized formulation, the current investigation is the first study on long-term sterilization effects of intratesticular injection of CaCl₂ in either lidocaine solution or alcohol in dogs. CaCl₂ at 20% concentration in lidocaine solution or alcohol was administered via intratesticular injection to groups of 21 dogs each. The treated animals were examined at 2, 6, and 12 months for sperm production, blood levels of testosterone, and side effects; at time zero and 12 months for testicular size and semen volume. The experimentally treated animals were compared to a control group receiving saline injection only.

Results

Testicles of dogs treated with CaCl₂ in either diluent significantly decreased in size. After administration of CaCl₂ in lidocaine solution, sterility was achieved for at least 12 months in 75% of treated dogs. However, optimal long-term contraceptive effectiveness was achieved with CaCl₂ in alcohol, which resulted in azoospermia over the 12-month study period. Testosterone levels significantly decreased following treatment with CaCl₂, and sexual activity disappeared. Although testosterone returned to baseline levels by 12 months for the group treated with CaCl₂ in lidocaine, dogs injected with CaCl₂ in alcohol had a 63.6% drop in testosterone level, which remained at the low end of physiological range throughout the study. No adverse effects were noted.

Conclusions

A single, bilateral intratesticular injection of 20% CaCl₂ in 95% ethanol was a reliable method for induction of sterilization in 18–28 kg male dogs in this study. The approach showed long-term efficacy and reduced sexual behavior. This chemical method of sterilization might provide an effective, efficient alternative to surgical castration that can have positive impacts on dog welfare.     

The resistance of meat chickens vaccinated by aerosol with a live V4 Newcastle disease virus vaccine in the field to challenge with a velogenic Newcastle disease virus

Meat chickens housed on a commercial broiler farm in Australia were vaccinated once at 10 to 11 days-of-age by aerosol with live V4 Newcastle disease virus (NDV) vaccine. Groups of vaccinated and unvaccinated birds were flown to Malaysia, where they were challenged with a virulent strain of NDV. Survival rates in vaccinated chickens challenged 7, 14, 21 or 31 d after vaccination were 0.47, 0.77, 0.97 and 0.92, respectively. All unvaccinated chickens died due to Newcastle disease (ND) following challenge. Chickens in Australia and Malaysia were bled and the serums tested for haemagglutination-inhibiting (HI) antibody to NDV. Many vaccinated birds with no detectable antibody, and all birds with a log2 titre of 2 or greater, survived challenge. The results showed that this V4 vaccine induced protective immunity in a significant proportion of chickens within 7 d of mass aerosol vaccination. This early immunity occurred in the absence of detectable circulating HI antibody. Non-HI antibody mediated immunity continued to provide protection up to 31 d after vaccination. Almost all vaccinated birds were protected within 3 w of vaccination. It is concluded that the V4 vaccine is efficacious and could be useful during an outbreak of virulent ND in Australia.     

The experimental production of big liver and spleen disease in broiler breeder hens

SUMMARY Big liver and spleen disease (BLS) was reproduced experimentally by intravenous (IV) and oral (PO) administration of BLS inocula to susceptible broiler breeder hens 34 to 36 weeks of age. Serological and pathological signs of BLS similar to those seen in the natural disease occurred in inoculated and in-contact birds. Splenomegaly was the earliest and often the only necropsy finding, with hepatomegaly and kidney enlargement occurring in some birds later in the course of the disease. After IV administration, serum antigen was detected between 2 and 4 weeks, and antibody between 3 and 5 weeks. After PO administration, antigen was detected between 2 and 4 weeks, and antibody between 3 and 6 weeks. Antibody persisted in all birds to the end of the experiment (6 weeks), and horizontal transmission probably occurred since in-contact birds developed BLS. Liver probably contained the highest concentration of BLS agent because it had the highest infectivity.     

Advances in Prostatic Diagnostics in Dogs: The Role of Canine Prostatic Specific Esterase in the Early Diagnosis of Prostatic Disorders

In last years, following the increased canine life expectancy and the rising attention pet-owners devote to their animals, several authors have carried on investigations concerning new techniques to early identify canine prostatic disorders that might affect the dog's quality of life. Prostatic disorders often have an asymptomatic onset and their early diagnosis is difficult: hence, they are usually identified at an advanced stage, only. Traditionally, the diagnosis of prostatic disorders is based on noninvasive tools, such as transrectal and abdominal palpation, seminal or prostatic fluid evaluation, and urinalysis and imaging. On the other hand, a definite diagnosis of prostatic abnormalities could be achieved through prostatic parenchyma Fine Needle Aspiration (FNA) or biopsy. However, these investigations are performed rarely because of their invasiveness. Thus, several authors investigated canine serum biomarkers in order to achieve an earlier diagnostic timing and to apply therapeutic strategies for better outcomes. The Canine Prostatic Specific Esterase (CPSE) has been identified as a suitable biomarker to be included in a prostate health screening program, following the model of prostate-specific antigen (PSA) in human medicine. A higher CPSE in dogs suffering from several prostatic diseases, such as benign prostatic hyperplasia, bacterial prostatitis, or prostatic carcinoma, was reported in literature. Thanks to the potential usefulness in clinical practice, further studies should investigate the potential role of CPSE in monitoring the medical treatment success in the male reproductive system. Moreover, the spreading availability of serum biomarkers, easily carried out on blood samples in clinical practice, could assure a more accurate evaluation of the actual prevalence of prostatic disorders. The CPSE is actually recognized as a promising diagnostic tool for the detection of prostatic disorders in a "prostate health screening program," in order to properly select those patients requiring further more accurate and expensive diagnostic investigations.