To evaluate the transplacental transfer of Theileria sergenti infection in cattle, we used DNA probes to detect T. sergenti in 6 pregnant cows and their calves. All the animals were monitored by parasitologic, serologic, and polymerase chain reaction (PCR) assays for a predicted 875-base-pair (bp) DNA product and a 684-bp amplicon detected by nested PCR in the blood and spleens of aborted fetuses. An open reading frame (ORF) starting at nucleotide 170 and terminating at position 1021 was shown to code for a polypeptide of 283 amino acid residues. All 6 dams and 5 calves were positive for T. sergenti in all tests. One calf was positive only with nested PCR. We conclude that transplacental transmission of T. sergenti is a significant problem. The relevance of the data in the programmed introduction of new (especially pregnant) animals into established clean herds needs serious consideration with regard to control of theileriosis and other tickborne diseases.

The purpose of this study was to collect initial data to determine the potential clinical usefulness of a 13C-aminopyrine demethylation blood test, and whether additional clinical investigation is warranted. Six dogs, initially suspected of having hepatic disease based on their history, physical examination, imaging studies, general laboratory parameters, or any combination of the above, were enrolled in the study. A baseline blood sample was collected, 2 mg/kg 13C-aminopyrine was administered intravenously, and another blood sample was collected 45 min afterwards. Carbon dioxide was extracted from the blood samples and analyzed using fractional mass spectrometry. Results from the 13C-aminopyrine demethylation blood test were compared to clinical data and histologic findings. Intravenous administration of 13C-aminopyrine leads to a decrease in the percent dose of 13C recovered from dogs with histologically confirmed liver disease. Based on our results, a full-scale investigation of the potential clinical usefulness of a 13C-aminopyrine demethylation blood test for assessment of hepatic function in dogs is warranted.

The time available to implement successful control measures against epidemics was estimated. Critical response time (CRT), defined as the time interval within which the number of epidemic cases remains stationary (so that interventions implemented within CRT may be the most effective or least costly), was assessed during the early epidemic phase, when the number of cases grows linearly over time. The CRT was calculated from data of the 2001 foot-and-mouth disease (FMD) epidemic that occurred in Uruguay. Significant regional CRT differences (ranging from 1.4 to 2.7 days) were observed. The CRT may facilitate selection of control measures. For instance, a CRT equal to 3 days would support the selection of measures, such as stamping-out, implementable within 3 days, but rule out measures, such as post-outbreak vaccination, because intervention and immunity building require more than 3 days. Its use in rapidly disseminating diseases, such as FMD, may result in regionalized decision-making.

Recently, a radioimmunoassay (RIA) for measurement of canine pancreatic lipase immunoreactivity (cPLI) in serum was developed and validated. However, RIAs require frequent use of radioactive materials. Therefore, the goal of this project was to develop and validate an enzyme-linked immunosorbent assay (ELISA) for cPLI. After purifying cPL, we developed and purified antiserum against cPL in rabbits. The purified antibody was bound to microtitre plates and used to capture antigen. A portion of the purified antibody was biotinylated and used to identify the captured antigen. Streptavidin labelled with horseradish peroxidase and a horseradish peroxidase substrate were used for detection. The assay was validated by determination of sensitivity, working range, linearity, accuracy, precision, and reproducibility. The reference interval for serum cPLI was determined by the central 95th percentile in 74 clinically healthy dogs: 2.2 to 102.1 μg/L. The sensitivity and the upper limit of the working range were 0.1 and 999.2 μg/L, respectively. The ratios of observed to expected values for dilutional parallelism for 6 serum samples ranged from 0.0 to 148.8%; the ratios for spiking recovery for 4 serum samples ranged from 90.4 to 112.6%, assuming 55% recovery of the cPL. Coefficients of variation for intra- and interassay variability for 6 different serum samples were 2.4, 3.4, 4.1, 5.8, 7.4, and 10.0% and 5.9, 7.7, 11.6, 13.9, 23.5, and 46.2%, respectively. We conclude that the ELISA described here is sufficiently sensitive, linear, accurate, precise, and reproducible for clinical application. Evaluation of its clinical usefulness for the diagnosis of exocrine pancreatic disorders in dogs is under way.

A polymerase chain reaction (PCR) assay was combined with a broth-culture enrichment system to detect Salmonella shed in feces from subclinically infected swine. The effectiveness of the broth culture-polymerase chain reaction (BC-PCR) assay to identify pigs shedding Salmonella in feces was compared with a microbiological culture and a commercial enzyme linked immunosorbent assay (ELISA) kit to detect Salmonella-specific serum antibody. A total of 67 pigs were tested by each of the 3 methodologies. Forty-one pigs tested positive for Salmonella by BC-PCR and ELISA identified 6 positives and 23 suspicious samples. It was shown that the BC-PCR assay is a rapid diagnostic tool for detecting of Salmonella shed by asymptomatic swine compared with current diagnostic technologies.

This study was conducted to ascertain anatomical variation in different body parts of Desi, Fayoumi, Cross (Rhode Island Red x Fayoumi) and Nick Chick laying hens. A total 16 laying hens of average body weight were selected using random numbers out of 2600 laying hens, slaughtered and eviscerated. It was observed that proportion of neck, ribs, breast, back, wings, thigh and legs out of aggregate weight was highest (51.74 plus minus 2.85) in Fayoumi hens. The proportion of liver, gizzard, heart and spleen combined weight was highest (6.05 plus minus 1.81) in desi hens. The proportion of non edible offals including trachea, lungs, kidneys, blood, feathers, head, crop, proventriculus, intestines, shanks, gizzard waste, skin, tail, testes, ova, oviduct and fat was estimated to be highest i.e. 46.60 plus minus 1.16 in Nick Chick laying hens. The anatomical variation in Desi and Fayoumi chicks was attributable to scavenging habit of these birds. The high proportion of edible offal in commercial Nick Chick hens may be ascribable to higher productive and reproductive traits.

This study was conducted to determine the optimum storage temperature for Rinderpest vaccine prepared on vero cells to know the shelf life of the vaccine. The vials were randomly selected from one batch of the vaccine, titrated and stored at minus 20 degree centigrade (Freezer), 4 degree centigrade (refrigerator) and room temperature. The titre was found to be 105.1 per ml. The vials stored at minus 20 degree centigrade & 4 degree centigrade were subjected to titration after an interval of six months for 3 and 2 years respectively. The vials stored at room temperature were tested after 4 weeks, 8 weeks and 12 weeks. Titration results indicated that the titre of the vaccine vials stored at room temperature decreased by 100.9 101.2 and 101.6 after storage time of 4 weeks, 8 weeks and 12 weeks respectively. The vials stored at 4 degree centigrade maintained their titre for a period of six months but after that the loss in titre was 100.4, 101.0, and 102.4 after storage time of one, one and half and two 2 years respectively. The vaccine vials stored at minus 20 degree centigrade maintained their original titres (initial titre of the vaccine) even after the storage for three years. It is concluded that vero cell adapted Rinderpest virus vaccine can be stored at 4 degree centigrade for a period of six months, however, at 20 degree centigrade it can be stored for three years without any adverse effect on titre.

The effects of melatonin and Light treatments on the reproductive performance of yearling Suffolk rams were investigated. The two groups (A, n = 8; B, n = 8) were given a priming period of long days (18 h Light (L): 6 h dark (D) during 1st February to 14th March. During 14th March to 9th September, group A was exposed to local light + melatonin Implants (Regulin) at 5 weekly intervals, group B was exposed to local light + melatonin Fed (M- 5250) alongwith the feed. The group (C, n = 8) in the local light during 1st February to 14th March and during 14th March to 9th September remained on local light + melatonin implants (Regulin) at 5-weekly intervals. However, the group (D, n=8) was treated as control and kept on local light environment (Lat. 51 degree 43'N) throughout the experiment. Testis diameter, sexual behaviours and semen quality were used to assess reproductive performances. The treatment to group A & B resulted in a significant advancement in high reproductive performance. The treatment to group C indicated that the abrupt application of melatonin in mid March resulted in a weak response in terms of all the criteria of assessment. The investigation indicated that seasonal fluctuations in reproductive behaviour and semen quality may be rephrased by the application of light and melatonin treatments and that these treatments may be considered for the preparation of rams for out of season breeding.

The quick diagnosis of Newcastle disease requires known serum against the disease. In this study, an attempt was made to raise anti- Newcastle disease virus hyperimmune serum in rabbits. Three different inoculum were prepared to inoculate in the rabbits; (i) Fresh harvested allantoic fluid containing Newcastle disease virus (NDV) Mukteswar vaccine strain; (ii) Freshly harvested ND virus pelletted through centrifuging at 40,000 rpm for two hours and resuspended in normal saline and (iii) Pelletted virus (centrifuged and suspended as in ii) with addition of incomplete Freund's adjuvant. It was observed during the monitoring of haemagglutination inhibition (HI) titre that the serum collected after series of inoculation of 1st and 2nd inoculum provided maximum titre upto 1:256. However, the serum collected after series of injection of 3rd inoculum gave maximum HI titre 1:1024. This study suggested that antigen containing incomplete Freund's adjuvant provided better immune response against Newcastle disease virus in rabbits.

This study was carried out for investigation of an outbreak in Karachi. A disease outbreak in poultry was reported during April, 2000 in Karachi. The main symptoms included respiratory distress, sneezing and gasping. Autopsy of dead birds showed tracheitis, air-sacculitis and involvement of lungs. The morbid material was collected for processing in the laboratory. During the investigation for isolation of pathogenic bacteria or virus, Ornithobacterium rhinotracheale was isolated on 5% sheep blood agar plates in an atmosphere containing 5-10% CO2 at 37 degree C and identified through biochemical and fermentation tests. A Lasota like virus was also isolated from the same material which showed HA activity but was found negative to Avian Influenza virus against known Avian Influenza serum. The isolated virus was sent to Central Veterinary Laboratory, Weybridge, U.K. which confirmed it to be Lasota virus having comparatively high Intracerebral Pathogenicity Index (ICPI) as 0.90. Attempts for the transmission of the disease in susceptible healthy broiler chicks was successful through aerosol route using combination of inoculum of the isolated bacteria and virus. On post-mortem of infected birds air- sacculitis, tracheitis and unilateral pneumonia were noticed. Again ORT and NDV were isolated from the material harvested from these chicks. According to the observations of these experiments, it was concluded that the isolated ORT had a triggering effect on Lasota virus or vice versa. The environmental temperature and other stress factors might have aggravated the disease problem.