Template-Type: ReDIF-Article 1.0 Author-Name: D. Rysanek Author-Workplace-Name: Veterinary Research Institute, Brno, Czech Republic Author-Name: V. Babak Author-Workplace-Name: Veterinary Research Institute, Brno, Czech Republic Author-Name: M. Zouharova Author-Workplace-Name: Veterinary Research Institute, Brno, Czech Republic Title: Bulk tank milk somatic cell count and sources of raw milk contamination with mastitis pathogens Abstract: The objective of this study was to probe the relationship between prevalence of selected principal mastitis pathogens and somatic cell counts in bulk tank milk samples. The sources of milk contamination were evaluated. The samples were collected from 298 dairy herds (with approximately 32 000 dairy cows). Only 48.3% of the bulk tank milk samples were free of contamination of pathogens of interest. Approximately 38.9% of the milk samples were contaminated with only one, 12.4% with two and 0.3% with three pathogens. The arithmetic mean of logarithmically transformed data of bulk tank milk somatic cell count rise in order: pathogen free, Pseudomonas aeruginosa, Streptococcus uberis, Escherichia coli and Staphylococcus aureus (5.381; 5.413; 5.495; 5.518; 5.563, respectively). The arithmetic mean differences between bulk tank milk somatic cell counts in pathogen-free and single-pathogen contaminated samples have revealed a significance for the Escherichia coli and Staphylococcus aureus groups (P < 0.01). Using binary logistic regression, a statistically highly significant relationship (P < 0.001) has been found between the number of contaminations of bulk tank milk samples with mastitis pathogens and bulk tank milk somatic cell counts. The relationship allows the determination of the probability of finding relevant mastitis pathogens in bulk tank milk samples with different levels of bulk tank milk SCC. A 63% probability can be defined at a cell count level of 400 000/ml and 20% at a cell count level of 100 000/ml. Analysis may reveal the potential sources of the bulk tank milk sample contamination, i.e. infected mammary glands or the environment. The presence of high levels of contamination along with a low bulk tank SCC may suggest an environmental source of contamination. The study clarified that a potential source of bulk tank milk contamination by relevant pathogens (the environment or the mammary gland) may be elucidated and the probability of the contamination of bulk tank milk samples with mastitis pathogens predicted by the analysis of relationship between the bulk tank milk somatic cell counts and the number of mastitis pathogen contaminations. Keywords: bulk tank milk samples, somatic cell count (SCC), Streptococcus uberis, Pseudomonas aeruginosa, Escherichia coli, Staphylococcus aureus Journal: Veterinární medicína Pages: 223-230 Volume: 52 Issue: 6 Year: 2007 DOI: 10.17221/1878-VETMED File-URL: http://vetmed.agriculturejournals.cz/doi/10.17221/1878-VETMED.html File-Format: text/html X-File-Ref: http://agriculturejournals.cz/RePEc/caa/references/vet-200706-0001.txt Handle: RePEc:caa:jnlvet:v:52:y:2007:i:6:id:1878-VETMED Template-Type: ReDIF-Article 1.0 Author-Name: C. Werner-Misof Author-Workplace-Name: Physiology Weihenstephan, Technical University Munich, Freising, Germany Author-Name: M.W. Pfaffl Author-Workplace-Name: Physiology Weihenstephan, Technical University Munich, Freising, Germany Author-Name: R.M. Bruckmaier Author-Workplace-Name: Veterinary Physiology, Vetsuisse Faculty, University of Bern, Bern, Switzerland Title: Dose-dependent immune response in milk cells and mammary tissue after intramammary administration of lipopolysaccharide in dairy cows Abstract: The immune response in milk cells and the status of mammary tight junctions (TJ) in response to intramammary (IM) infusion of different doses of Escherichia coli lipopolysaccharide (LPS) was investigated. Experiment I: Seven German Braunvieh cows were IM infused into one quarter with 1 μg (LPS-1) and 3 μg (LPS-3) of LPS, respectively, and the contralateral control quarter with saline (9 g/l; C). Milk samples were taken immediately before and 12, 24, 36, 48, 60, 84 and 108 h after infusion and analysed for somatic cell counts (SCC), lactose, sodium (Na) and chloride (Cl) ions, and electrical conductivity (EC). Milk cell mRNA expression of various inflammatory factors was quantified by real-time RT-PCR. Blood samples were taken immediately after milking for the analysis of leukocytes (WBC), polymorphonuclear neutrophils (PMN), Na and Cl. Milk SCC, lactose, Na, Cl and EC did not differ significantly between LPS-1 and C quarters after the challenge. In LPS-3 quarters SCC levels increased within the first 12 h, reached peak levels between 12 and 36 h (P ≤ 0.001) and decreased (P ≤ 0.05) thereafter to reach baseline at 108 hours. Lactose in LPS-3 quarters decreased (P ≤ 0.05) to a minimum at 24 h and increased slightly thereafter while EC, Na, and Cl increased transiently in response to LPS-3. WBC and PMN levels in both groups decreased numerically within 24 h after LPS administration. In LPS-1, WBC at 24, 48 and 108 h were significantly lower whereas in LPS-3 they were significantly higher than at time 0. TNFα-mRNA expression in both groups did not change in response to IM LPS-challenge. IL-1β-mRNA expression at 12, 24 and 36 h in LPS-1 quarters increased significantly as compared to time 0. In LPS-3 quarters the mRNA expression values of all tested ILs increased significantly as compared to time 0 within 12 h after LPS-challenge. IL-1β-mRNA expression decreased (P ≤ 0.05) at 48 and 84 h in LPS quarters. IL-8 mRNA was significantly decreased at 84 h after challenge in LPS-3 quarters. COX-2-mRNA expression in LPS-1 quarters decreased significantly as compared to time 0 at 48, 84 and 108 h, with a minimum at 84 h (P ≤ 0.05). In LPS-3 quarters COX-2-mRNA levels increased (P ≤ 0.05) within 48 h after the LPS-challenge. Experiment II: Six cows (5 German Braunvieh, 1 Brown Swiss) were injected in one quarter with 100 μg LPS and in the contralateral quarter with saline (9 g/l; C). Mammary biopsy samples of both quarters were taken immediately before and at 3, 6, 9 and 12 h after infusion and mRNA expression of TJ proteins occludin (OCLN) and zonula occludens (ZO-) 1, 2 and 3 were quantified by real-time RT-PCR. OCLN-mRNA expression did not change in response to the IM infusion while that of ZO-1, ZO-2 and ZO-3 decreased significantly within six hours. In conclusion, a dose of 1 μg LPS did not initiate a immune response in the mammary gland. Furthermore the dose of 100 μg of LPS enhanced TJ permeability by reducing TJ plaque proteins density. Keywords: mammary gland, mastitis, lipopolysaccharide, cattle Journal: Veterinární medicína Pages: 231-244 Volume: 52 Issue: 6 Year: 2007 DOI: 10.17221/1877-VETMED File-URL: http://vetmed.agriculturejournals.cz/doi/10.17221/1877-VETMED.html File-Format: text/html X-File-Ref: http://agriculturejournals.cz/RePEc/caa/references/vet-200706-0002.txt Handle: RePEc:caa:jnlvet:v:52:y:2007:i:6:id:1877-VETMED Template-Type: ReDIF-Article 1.0 Author-Name: S. Hiss Author-Workplace-Name: Institute of Animal Science, University of Bonn, Bonn, Germany Author-Name: U. Mueller Author-Workplace-Name: Institute of Animal Science, University of Bonn, Bonn, Germany Author-Name: A. Neu-Zahren Author-Workplace-Name: Institute of Animal Science, University of Bonn, Bonn, Germany Author-Name: H. Sauerwein Author-Workplace-Name: Institute of Animal Science, University of Bonn, Bonn, Germany Title: Haptoglobin and lactate dehydrogenase measurements in milk for the identification of subclinically diseased udder quarters Abstract: Diagnosis of subclinical mastitis is of increasing importance and appropriate detection methods are needed. Both haptoglobin (Hp), an acute phase protein in cattle, as well as lactate dehydrogenase (LDH), an ubiquitous enzyme, can be successfully used to detect clinical mastitis. The present paper describes quantification of Hp and LDH in milk samples from healthy and subclinically diseased udder quarters. Hp was analysed in the laboratory using an ELISA. The activity of LDH was measured in raw milk directly in the milking parlor. Both parameters were suitable to distinguish between sterile samples and bacteriologically positive samples. The ability to differentiate between minor and major pathogens was better for Hp in skim milk than for LDH in raw milk. Hp and somatic cell count (SCC) as well as LDH and SCC were positively correlated (r = 0.8 and r = 0.76, respectively). Subclinical mastitis was defined as follows: SCC > 100 × 103 cells/ml and bacteriological positive findings in two out of three weekly samples. Sensitivity and specificity were above 85% for Hp and above 81% for LDH. Using a less rigid classification to define mastitis, i.e. SCC < 200 × 103 cells/ml and two out of three weekly samples bacteriologically positive, sensitivity for Hp improved (89%) and remained unchanged for LDH. Both parameters are useful parameters for the diagnosis of subclinical mastitis. LDH activity in raw milk was less sensitive and specific than Hp but the method described herein offers the opportunity to measure LDH activity directly in the milking parlor and might therefore be suitable for on-line system developments. Keywords: haptoglobin, LDH, milk, diagnosis of subclinical mastitis Journal: Veterinární medicína Pages: 245-252 Volume: 52 Issue: 6 Year: 2007 DOI: 10.17221/1879-VETMED File-URL: http://vetmed.agriculturejournals.cz/doi/10.17221/1879-VETMED.html File-Format: text/html X-File-Ref: http://agriculturejournals.cz/RePEc/caa/references/vet-200706-0003.txt Handle: RePEc:caa:jnlvet:v:52:y:2007:i:6:id:1879-VETMED Template-Type: ReDIF-Article 1.0 Author-Name: E. Kosinova Author-Workplace-Name: Veterinary Research Institute, Brno, Czech Republic Author-Name: I. Psikal Author-Workplace-Name: Veterinary Research Institute, Brno, Czech Republic Author-Name: B. Robesova Author-Workplace-Name: Veterinary Research Institute, Brno, Czech Republic Author-Name: K. Kovarcik Author-Workplace-Name: Veterinary Research Institute, Brno, Czech Republic Title: Real-time PCR for quantitation of bovine viral diarrhea virus RNA using SYBR Green I fluorimetry Abstract: Quantitative real-time RT-PCR (qRT-PCR) assay was developed for the detection and quantification of bovine viral diarrhea virus (BVDV) in clinical samples from persistently infected cattle. qRT-PCR was optimized to quantify the number of BVD virus copies using Light Cycler® detection system and intercalation fluorogenic dye SYBR Green I. A universal set of primers was selected from a highly conserved 5' untranslated region (5'UTR) to detect BVDV type I and II simultaneously. Quantification of BVDV cDNA was accomplished using a calibration curve generated from 10-fold serial dilutions of standard plasmid DNA in the range 1-108 copies/μl. Analysis of 290 bp amplicons enabled monitoring of the viral RNA/BVDV level in a total of five BVDV strains (BVD-NADL, A03/3004, DB03/2943, KA04/3124, KV05/3412) and sixteen bulk milk samples, and in bovine sera of persistent carriers originating from Czech farms, as well as in a batch of calf serum for cell culture. Melting temperatures of amplicons (Tm) of BVDV strains of the same genotype group I as the NADL reference strain showed variability of the thermal points, however significant differences were observed in Tm values between the representatives of genotype group I and II. Low concentrations of BVD virus in bulk milk samples were also qualitatively identified by conventional RT-PCR. Highly reproducible data were obtained as the coefficients of variation of threshold cycles values in intra-assay and inter-assay were less than 0.85% and 2.76%, respectively. The results give enough evidence of suitability of qRT-PCR assay for quantitative analysis of BVDV in clinical samples. Keywords: bovine viral diarrhea virus, RNA, real-time RT-PCR, SYBR Green I Journal: Veterinární medicína Pages: 253-261 Volume: 52 Issue: 6 Year: 2007 DOI: 10.17221/1882-VETMED File-URL: http://vetmed.agriculturejournals.cz/doi/10.17221/1882-VETMED.html File-Format: text/html X-File-Ref: http://agriculturejournals.cz/RePEc/caa/references/vet-200706-0004.txt Handle: RePEc:caa:jnlvet:v:52:y:2007:i:6:id:1882-VETMED Template-Type: ReDIF-Article 1.0 Author-Name: E. Voslarova Author-Workplace-Name: University of Veterinary and Pharmaceutical Sciences, Brno, Czech Republic Author-Name: B. Janackova Author-Workplace-Name: Ministry of Agriculture of the Czech Republic, Prague, Czech Republic Author-Name: F. Vitula Author-Workplace-Name: University of Veterinary and Pharmaceutical Sciences, Brno, Czech Republic Author-Name: A. Kozak Author-Workplace-Name: Regional Veterinary Administration of the Capital Prague, Czech Republic Author-Name: V. Vecerek Author-Workplace-Name: University of Veterinary and Pharmaceutical Sciences, Brno, Czech Republic Title: Effects of transport distance and the season of the year on death rates among hens and roosters in transport to poultry processing plants in the Czech Republic in the period from 1997 to 2004 Abstract: Poor welfare is the cause of high mortality among hens and roosters transported to poultry processing plants. In the Czech Republic, death rates among hens and roosters in transport to poultry slaughter plants were monitored between 1997 and 2004, and their total mortality rate was in the 0.925% ± 0.479% range. Death rates among hens and roosters were influenced by the transport distance to poultry processing plants. The percentage of dead birds increased from 0.592% ± 0.575% at transport distances up to 50 km to 1.638% ± 0.952% at transport distances up to 300 km. The bird mortality was also influenced by the season of the year. Higher mortality rates were ascertained during the cold months of the year, specifically in October through to April. Keywords: welfare, stress, poultry, mortality, transportation time, ambient temperature, winter months Journal: Veterinární medicína Pages: 262-266 Volume: 52 Issue: 6 Year: 2007 DOI: 10.17221/1881-VETMED File-URL: http://vetmed.agriculturejournals.cz/doi/10.17221/1881-VETMED.html File-Format: text/html X-File-Ref: http://agriculturejournals.cz/RePEc/caa/references/vet-200706-0005.txt Handle: RePEc:caa:jnlvet:v:52:y:2007:i:6:id:1881-VETMED Template-Type: ReDIF-Article 1.0 Author-Name: E. Ludvikova Author-Workplace-Name: Equine Clinic, Faculty of Veterinary Medicine, University of Veterinary and Pharmaceutical Sciences, Brno, Czech Republic Author-Name: P. Jahn Author-Workplace-Name: Equine Clinic, Faculty of Veterinary Medicine, University of Veterinary and Pharmaceutical Sciences, Brno, Czech Republic Author-Name: Z. Lukas Author-Workplace-Name: Department of Pathology, Faculty of Medicine, Masaryk University, Brno, Czech Republic Title: Nutritional myodegeneration as a cause of dysphagia in adult horses: three case reports Abstract: Three cases of nutritional myodegeneration caused by selenium deficiency in adult horses are described. Difficulty in eating and drinking was a common clinical sign in all horses. Blood biochemistry revealed a marked elevation of muscle enzymes and low glutathione peroxidase activity or low selenium concentration in whole blood in all cases. The treatment with sodium selenite and vitamin E was instituted in all horses. Two of them were euthanized because of continuing muscle injuries, one patient was cured. The post-mortem examination of euthanized horses revealed pale muscles that were distributed with bilateral symmetry on hind and thoracic limbs, diaphragm, tongue, masticatory and intercostal muscles and the myocardium. Histopathology revealed the areas of degeneration and necrosis. Large groups of regenerating fibres and pronounced lymphoplasmocytic reaction among the groups of intact fibres were also present. The clinical outcome of the disease is probably influenced by timely diagnosis and treatment. Keywords: selenium, α -tocopherol acetate, vitamin E, myopathy, glutathione peroxidase Journal: Veterinární medicína Pages: 267-272 Volume: 52 Issue: 6 Year: 2007 DOI: 10.17221/1880-VETMED File-URL: http://vetmed.agriculturejournals.cz/doi/10.17221/1880-VETMED.html File-Format: text/html X-File-Ref: http://agriculturejournals.cz/RePEc/caa/references/vet-200706-0006.txt Handle: RePEc:caa:jnlvet:v:52:y:2007:i:6:id:1880-VETMED