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A quantitative real-time PCR assay for Ehrlichia ruminantium using pCS20 |
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Written by Administrator
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Sunday, 01 June 2008 |
Vet Microbiol. 2008 May 27;
Authors: Steyn HC, Pretorius A, McCrindle CM, Steinmann CM, Van Kleef M
Heartwater is a tick borne disease that affects ruminants and wild animals in Africa south of the Sahara. It is caused by Ehrlichia ruminantium and transmitted by the tick Amblyomma hebraeum. The protocols currently used to detect heartwater take several days to complete. Here, we describe the development of a pCS20 quantitative real-time PCR TaqMan probe assay to detect E. ruminantium in livestock blood and ticks from the field. The assay is based on the conserved pCS20 gene region of E. ruminantium that contains two overlapping genes, rnc and ctaG [Collins, N.E., Liebenberg, J., De Villiers, E.P., Brayton, K.A., Louw, E., Pretorius, A., Faber, F.E., Van Heerden, H., Josemans, A., Van Kleef, M., Steyn, H.C., Van Strijp, M.F., Zweygarth, E., Jongejan, F., Maillard, J.C., Berthier, D., Botha, M., Joubert, F., Corton, C.H., Thomson, N.R., Allsopp, M.T., Allsopp, B.A., 2005. The genome of the heartwater agent Ehrlichia ruminantium contains multiple tandem repeats of actively variable copy number. PNAS 102, 838-843]. The pCS20 quantitative real-time PCR TaqMan probe was compared to the currently used pCS20 PCR and PCR/(32)P-probe test with regards to sensitivity, specificity and the ability to detect DNA in field samples and in blood from experimentally infected sheep. This investigation showed that the pCS20 quantitative real-time PCR TaqMan probe was the most sensitive assay detecting seven copies of DNA/mul of cell culture. All three assays, however, cross react with Ehrlichia canis and Ehrlichia chaffeensis. The pCS20 real-time PCR detected significantly more positive field samples. Both the PCR and pCS20 real-time PCR could only detect E. ruminantium parasites in the blood of experimentally infected sheep during the febrile reaction. The PCR/(32)P-probe assay, however, detected the parasite DNA 1 day before and during the febrile reaction. Thus, because this new quantitative pCS20 real-time PCR TaqMan probe assay was the most sensitive and can be performed within 2h it is an effective assay for epidemiological surveillance and monitoring of infected animals.
PMID: 18511221 [PubMed - as supplied by publisher] |
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Prototheca zopfii genotypes isolated from cow barns and bovine mastitis in Japan |
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Written by Administrator
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Sunday, 01 June 2008 |
Vet Microbiol. 2008 May 27;
Authors: Osumi T, Kishimoto Y, Kano R, Maruyama H, Onozaki M, Makimura K, Ito T, Matsubara K, Hasegawa A
This study is the first investigation on Japanese isolates of Prototheca zopfii from bovine mastitis and the cow-barn surroundings by molecular characterization to clarify routes of infection for bovine protothecal mastitis. We performed isolation of Prototheca from cow-barn surroundings (drinking water, sewage and feces) and milk samples from cases of bovine mastitis. Genotypes of the 32 isolates of P. zopfii from cow-barn surroundings and 67 isolates from mastitis were analyzed by genotype-specific PCR assays and restriction fragment length polymorphism (RFLP) assays. All mastitis isolates were identified as P. zopfii genotype 2. Conversely, 29 isolates from cow-barn surroundings were identified as P. zopfii genotypes 1 and 3 isolates as genotype 2, respectively. Given these results, both genotypes of P. zopfii could exist in cow-barn surroundings, but no sites were identified as frequent sources of P. zopfii genotype 2. P. zopfii isolates should thus be further explored with regard to genotype to clarify the reservoir of etiological agents in bovine Prototheca mastitis.
PMID: 18511222 [PubMed - as supplied by publisher] |
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Prevalence of gastric ulcers in Thoroughbred broodmares in pasture: A preliminary report |
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Written by Administrator
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Sunday, 01 June 2008 |
Vet J. 2008 May 27;
Authors: le Jeune SS, Nieto JE, Dechant JE, Snyder JR
Gastroscopic examinations were performed in 62 Thoroughbred broodmares (33 pregnant, 29 non-pregnant) at one breeding farm to investigate the prevalence of gastric ulceration. Age, pregnancy status, race earnings, last race start, herd size, medical history, number of live foals, breeding years, feed type and number of feedings were recorded, plus coat condition and body condition score were determined. Twenty-one mares were re-evaluated after foaling, and the foaling date, foal weight at birth and placenta weight were recorded. The overall prevalence of gastric ulcers was 70.9%, with a median ulcer score of 3.0 (range: 2-5). Most ulcers were present on the squamous portion of the stomach, while two mares had glandular ulcers. There were no differences in the presence, location and severity of gastric ulcers between pregnant and non-pregnant mares. Furthermore, there were no significant associations between the variables measured and the presence of gastric ulceration. The prevalence of gastric ulceration in this specific population of horses was higher than expected and further investigation is warranted to determine the factors that contributed to this finding.
PMID: 18511313 [PubMed - as supplied by publisher] |
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Interference of iohexol with radioiodine thyroid uptake in the hyperthyroid cat |
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Written by Administrator
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Sunday, 01 June 2008 |
Feline Med Surg. 2008 May 27;
Authors: Peremans K, Vandermeulen E, van Hoek I, Daminet S, Vermeire S, Bacher K
Absorbed thyroid dose and effective half-life were determined in 46 hyperthyroid cats after treatment with a low dose (mean 111MBq) of radioiodine intravenously. Thirteen of these cats had received iohexol for glomerular filtration rate (GFR) measurement within 24h before treatment with radioiodine in view of another ongoing study at our institution. Pre-therapy values were obtained for total thyroxine (TT(4)) and for the thyroid to salivary gland ratio with sodium pertechnetate gamma-camera imaging. All cats underwent post-therapy scans at 24, 48 and 120h for evaluation of radioactive iodine uptake (RAIU) and the effective half-life of radioiodine. The absorbed dose was calculated from the cumulative activity with Olinda software. Both groups were comparable in age, TT(4) and the ratio of thyroid activity to salivary gland activity. Statistical analysis revealed a significant decreased absorbed dose in the thyroid in the iohexol group. This decreased uptake was not accompanied by an decreased effective half-life of the radioiodine. The variation of inter-individual RAIU decreased in this group and more homogenous absorbed doses were obtained. No significant difference in outcome could be demonstrated. However, a tendency towards a higher number of residual hyperthyroidism in the iohexol group was noted (15 versus 6% in control group). This study demonstrates that iohexol interferes with the uptake of radioiodine in the hyperthyroid cat but does not provoke increased turnover. In this study, albeit including a small number of cats, outcome did not seem to be significantly affected.
PMID: 18511320 [PubMed - as supplied by publisher] |
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Echocardiographic measurements of cardiac dimensions in normal Standardbred racehorses |
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Written by Administrator
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Sunday, 01 June 2008 |
J Vet Cardiol. 2008 May 27;
Authors: Zucca E, Ferrucci F, Croci C, Di Fabio V, Zaninelli M, Ferro E
OBJECTIVES: The aim of this study was to obtain echocardiographic measurements and establish reference ranges for 14 parameters in Standardbred racehorses in training. BACKGROUND: Several studies have been published about cardiac measurements in Thoroughbreds, Standardbreds, National Hunt horses, Warmbloods and ponies; however, not all parameters have been published for the Standardbred trotter in training. ANIMALS, MATERIALS AND METHODS: Thirty normal Standardbred racehorses in training were assessed by two-dimensional echocardiography (2-D) and M-mode echocardiography using standardized imaging planes. Mean values, standard deviations, 95% confidence interval for the means and 95% confidence interval for the cardiac parameters measured in the population were calculated. Furthermore, a general linear model was constructed using sex, age and body weight (bwt) of the horses as independent variables and the echocardiographic measurements as dependent variables. Multivariate linear regression analysis was performed with the level of significance at p<0.05 for all the null hypotheses. RESULTS: Reference ranges were established for 14 echocardiographic parameters in Standardbred racehorses. Weak linear relationships between echocardiographic measurements and body weight were observed for LVIDd, LVIDs, LVFWs, and AOD. Linear regressions on these parameters were used to calculate the 95% confidence intervals for the predicted values. CONCLUSIONS: The data collected in this study provide reference values for the evaluation of Standardbred racehorses in training. Body weight has a negligible affect on most echocardiographic parameters in this homogeneous population, but did mildly influence the results of left ventricular and aortic measurements.
PMID: 18511359 [PubMed - as supplied by publisher] |
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