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Development and analytic validation of an enzyme-linked immunosorbent assay for the measurement of canine pancreatic lipase immunoreactivity in serum
2003
Steiner, Jörg M. | Teague, Sheila R. | Williams, David A.
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.
Mostrar más [+] Menos [-]Comparison of bacterial enriched-broth culture, enzyme linked immunosorbent assay, and broth culture-polymerase chain reaction techniques for identifying asymptomatic infections with Salmonella in swine
2003
Sibley, Jennifer | Yue, Binbin | Huang, Fei | Harding, John | Kingdon, Jill | Chirino-Trejo, Manuel | Appleyard, Grey D.
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.
Mostrar más [+] Menos [-]General morphology of the oral cavity of the Nile crocodile, <i>Crocodylus niloticus<i/> (Laurenti, 1768). I. Palate and gingivae
2003
J.F. Putterill | J.T. Soley
The heads of nine 2.5 to 3-year-old Nile crocodiles (Crocodylus niloticus) were obtained from a commercial farm where crocodiles are raised for their skins and meat. The animals from which these specimens originated were clinically healthy at the time they were slaughtered. A detailed description of the macroscopic and microscopic features of the palate and gingivae of the Nile crocodile is presented and the results are compared with published information on this species and other Crocodylia. The histological features are supplemented by information supplied by scanning electron microscopy. Macroscopic features of interest are the small conical process situated at the base of the first two incisors of the maxilla, the distribution of cobbled units on the palate, and the broad dentary shelf forming the rostral aspect of the mandible. Histologically the palate and gingivae did not differ significantly from each other and both regions showed a presence of Pacinian-type corpuscles. Two types of sensory structures (taste receptors and pressure receptors) were identified in the regions examined, both involving modification of the epithelium and the underlying connective tissue.
Mostrar más [+] Menos [-]Molecular analysis of bovine viral diarrhoea virus isolates from South Africa
2003
N. Kabongo | C. Baule | M. Van Vuuren
The presence of bovine viral diarrhoea virus in South Africa has been confirmed by several serological surveys. However, little is known about its biological properties. Twenty five isolates obtained by isolation in tissue culture and detected by means of the antigen capture ELISA from clinically sick cattle and from foetal calf serum in South Africa were characterized on the basis of analysis of the 5' non-translated (NTR) region of the genome. A reverse-transcription polymerase chain reaction (RT-PCR) was used to amplify specific sequences from the 5'NTR of the genome. The oligonucleotide primers corresponding to positions 105-125 and 399-378, respectively, in the sequence of BVDV strain NADL were used to generate the PCR products. Both strands were sequenced directly with these primers and fluorescence-labelled dideoxynucleotides in an automated nucleic acid sequencer. Reference strains of pestiviruses [(BVDV type I, BVDV type II, border disease virus (BDV) and hog cholera virus (HCV)] and isolates from a previous investigation on BVDV in southern Africa were included for comparative purposes. All the BVDV strains obtained during this study belong to subgroups of BVDV genotype I. No association could be demonstrated between the geographic origin of the isolates. A number of isolates formed another branch separate from the existing branches Ia, Ib and Ic. These findings suggest that extensive genetic diversity can be found within BVDV type I isolates from southern Africa. Isolates that group with the classical BVDV type I strains, particularly of American origin, coexist with variants that appear to represent a local genetic pool and or variants evolving from the classical strains.
Mostrar más [+] Menos [-]The heart remembers : observations of cardiac memory in the Dorper sheep heart
2003
J. Ker | E.C. Webb | J.A. Ker | P.A. Bekker
Memory is a property common to a diverse range of tissues. Cardiac memory has been demonstrated in the human, dog, rat and rabbit. This is a peculiar phenomenon, reflected in the T wave of the electrocardiogram. The heart is able to remember periods of alterations in the sequence of ventricular activation and once there is a return to a normal sequence of ventricular activation the T waves may manifest memory. Cardiac memory is noted when the T wave during normal ventricular activation retains the vector of the previous abnormal QRS complex, caused by a period of altered ventricular activation. Possible mechanisms of memory in the heart are alterations of the transient outward potassium current (Ito) in ventricular myocytes and new protein synthesis inside myocytes. These two mechanisms operate in short- and long-term cardiac memory respectively. Currently, it is unknown whether memory may have adverse structural consequences in the heart. We were able to demonstrate memory in the hearts of Dorper wethers and this is the first report of cardiac memory in Dorper sheep.
Mostrar más [+] Menos [-]Review of blackfly (Diptera : Simuliidae) control in South Africa
2003
E. Myburgh | E.M. Nevill
The medical, veterinary and economic importance of blackflies in South Africa, and the historical development of blackfly control programmes in various South African rivers, are reviewed in this paper. In 1996 it was estimated that blackflies can cause more than R 88 million damages per annum along the middle and lower Orange River where Simulium chutteri is considered the main pest species. A clear link between the construction of dams and the spread of the blackfly problem was shown. Four phases characterize the development of blackfly control in South Africa: (1) during the 1960s blackflies in the Vaal River were controlled with DDT; (2), during the 1970s and into the 1980s blackflies were controlled using water-flow manipulation; (3) when used at strategic times, water-flow manipulation could be used to enhance the effect of natural predator populations; and (4) during the 1990s the organophosphate temephos and toxins produced by the bacterium Bacillus thuringiensis var. israelensis were tested for their efficacy against blackflies. The larvicides temephos and B. thuringiensis proved to be effective and are still used in several control programmes. The latest research focuses on the factors that influence adult blackfly survival and annoyance, as well as the development of methods that can be used to protect sheep from blackfly attacks.
Mostrar más [+] Menos [-]Population structure of <i>Argas arboreus<i/> (Acari : Argasidae) ticks associated with seasonally abandoned mixed heronries, dominated by cattle egrets (<i>Bubulcus ibis<i/>), in South Africa
2003
V.N. Belozerov | D.J. Van Niekerk | H.J.B. Butler
During winter populations of Argas arboreus from heronries of the cattle egret, Bubulcus ibis, in South Africa are composed of adults, with some predominance of males, and II-IV instar nymphs, in a state of diapause. The period of tick activity, including reproduction and development of eggs, larvae and N1 nymphs, is synchronized with the nesting and breeding season of their avian hosts. It begins during spring with the return of birds to the heronry, and ceases in autumn through induction of reproductive diapause in engorged females, and behavioural diapause in unfed nymphs and adult ticks. Many ticks showed morphological anomalies and malformations, the study of which could possibly be used for monitoring of environmental pollution.
Mostrar más [+] Menos [-]Kinetics and postmucosal effects on urinary recovery of 5 intravenously administered sugars in healthy cats
2003
Krecic, Matthew R. | Steiner, Jörg M. | Kern, Margaret R. | Williams, David A.
The objective of this study was to describe the kinetics of urinary recovery and to evaluate the effects of postmucosal factors on urinary recovery of 5 intravenously administered saccharides. Ten cats received an isotonic sugar solution containing lactulose, rhamnose, xylose, methylglucose, and sucrose intravenously. These sugars were selected because of their prior use for intestinal permeability and mucosal function testing in humans and dogs. Urethral catheterization with a closed collection system was used for collection of cumulative urine samples prior to and 2, 4, 6, 8, 10, 12, and 24 h after administration of the sugar solution. High-pressure anion exchange liquid chromatography with pulsed amperometric detection was used to measure the concentrations of each sugar in the urine and calculate urinary recovery. Twenty-four hour cumulative urinary recovery for each sugar from the cats, was lower than expected compared to dogs and humans. All 5 sugars had the highest percentage of urinary recovery during the first 2 h after administration. Mean sugar elimination rate constants and half-lives ranged from 0.268/h for methylglucose to 0.415/h for lactulose and 1.67 h for lactulose to 2.59 h for methylglucose, respectively. Metabolism and incomplete urine collection are possible reasons for lower cumulative urinary recoveries of these 5 sugars in cats compared with dogs. Although these 5 sugars are not ideal marker molecules, they may still be useful for intestinal permeability and mucosal function testing in cats.
Mostrar más [+] Menos [-]Experimental infection of pregnant gilts with swine hepatitis E virus
2003
Kasorndorkbua, Chaiyan | Thacker, Brad J. | Halbur, Patrick G. | Guenette, Denis K. | Buitenwerf, Ryan M. | Royer, Ryan L. | Meng, Xiang-Jin
To determine the effect of swine hepatitis E virus (HEV) infection on pregnant gilts, their fetuses, and offspring, 12 gilts were intravenously inoculated with swine HEV. Six gilts, who were not inoculated, served as controls. All inoculated gilts became actively infected and shed HEV in feces, but vertical transmission was not detected in the fetuses. There was no evidence of clinical disease in the gilts or their offspring. Mild multifocal lymphohistiocytic hepatitis was observed in 4 of 12 inoculated gilts. There was no significant effect of swine HEV on fetal size, fetal viability, or offspring birth weight or weight gain. The offspring acquired anti-HEV colostral antibodies but remained seronegative after the antibodies waned by 71 days of age. Swine HEV infection induced subclinical hepatitis in pregnant gilts, but had no effect on the gilts' reproductive performance, or the fetuses or offspring. Fulminant hepatitis associated with HEV infection was not reproduced in gilts.
Mostrar más [+] Menos [-]Ribotype characterization and antimicrobial susceptibility profiles of Campylobacter coli isolates from swine
2003
Sheffield, C.L. | Hume, M.E. | Droleskey, R.E. | Harvey, R.B. | Bischoff, K.M.
Objective: To evaluate the genetic relatedness and antibiotic sensitivity profiles of Campylobacter coli isolates from sows and piglets housed in an integrated swine production facility. Sample Population: Ninety-nine isolates of Campylobacter coli were collected from 3 sows (Yorkshire-Landrace) and 18 piglets (Yorkshire-Landrace X Duroc or Hampshire) housed in a common farrowing barn. Procedure: When piglets were weaned (21 day of age) fecal samples were collected for the sows and rectal samples were collected from the piglets. Isolation of Campylobacter coli was performed using an enrichment broth and restrictive media under microaerophilic conditions. Results: The Campylobacter coli isolates segregated into 20 ribogroups and exhibited 32 antibiotic susceptibility profiles. The Ribogroup (224-373-S-5) contained 35 isolates from eleven animals. Thirty-eight percent of the animals exhibited a single ribogroup, while 10% of the animals exhibited four ribogroups. No discernible pattern of ribogroup relatedness was observed among the sows and piglets or among littermates. Conclusion: The data suggests a high level of diversity in both ribotypic patterns and antibiotic sensitivity profiles among the Campylobacter coli isolated from related pigs housed in a single facility. Further, no evidence was found for a direct transfer of specific Campylobacter coli ribotypes from a sow to her piglets.
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