The humeral immune response of swine after experimental infection with Salmonella typhimurium

LPS antibody reaction of S. Typhimurium was detected in a total of 111 two- to ten-week-old pigs, which had been exposed to experimental oral infection with this serovar in 16 separate experiments. The total antibody level was determined according to the meat juice ELISA provided for the Salmonella control programme and the antibody concentrations of the isotopes IgG1, IgG2, IgA and IgM. Although there were significant differences between the individual experiments with respect to the reaction intensity and proportion of the individual immune globulin isotypes, the following general statements on the dynamics of the antibody reaction can be made: Clearly elevated antibody levels could be observed one week after the onset of infection. After one and two weeks, respectively, 22.5 and 52.9% of the infected animals showed antibody levels which were to be evaluated as positive in the sense of the Salmonella control programme (> or = 40%). While the concentration of the IgM-related antibodies decreased again during the second week already, the antibody concentration of IgG1 and IgG2 continued to increase until the end of the experiment after 3 weeks. The clear differences in the reactions between the experiments cannot be explained solely by defined conditions of infection such as strain of infection, infective dose or age of the pigs. Obviously, other non-defined or non-recognized factors may have a decisive influence on the dynamics of the antibody reaction in the animal groups.     

Hypofractionated radiation therapy for the treatment of malignant melanoma and squamous cell carcinoma in dogs and cats

This study describes the experience with hypofractionated radiation therapy of squamous cell carcinoma and melanoma in dogs and cats. A total dose of 32-48 Gray (Gy) was delivered once a week in 8 Gy fractions. 34 animals in which a complete surgical excision was impossible were treated. There was no tumor detectable macroscopically in 14 patients at the beginning of radiation therapy. In 20 animals the median volume of the tumor was 9.9 cm3. The median survival times and the local tumor control of squamous cell carcinoma of the oral and nasal cavities and of the body are comparable to results which were reached with a Monday-Wednesday-Friday scheme. For the treatment of Melanoma the hypofractionated radiation therapy is first choice. There are no significant side effects. Late side effects did not occur. 88% of the owners are satisfied with this kind of treatment and would choose it again.     

Usefulness of serological examinations in the analysis of Salmonella infections in pig herds

The development of the antibody concentration against lipopolysaccharide (LPS) of S. Typhimurium und S. Choleraesuis in rearing pigs during the fattening period and in breeding sows of the corresponding age was recorded. The studies revealed the following results. Antibodies of isotypes IgG1 and IgG2 revealed a more pronounced specificity against the according Salmonella serovar than IgM antibodies. The calculated "antibody percent value" based on the total amount of Salmonella antibodies is mainly determined by the IgM antibodies in sera and meat juice, respectively. In fattening pigs a significant increase of antibodies against IgM and total Ig was observed between week 3 and 10 after beginning of the rearing period. In breeding pigs this increase was detectable already earlier. In only 3 out of 10 groups an increase of IgG1 and IgG2 was also seen. The detected significant increase of total Ig and IgM in the other groups might be the result of a less intensive exposure to salmonellas or it might be due to an increase of unspecific antibodies induced by other antigens. Serological investigations represent a valuable tool to record the intensity and development in time of the Salmonella exposure in pigs farms. Examination of total Ig is an appropriate method to detect pig herds with a high level of Salmonella exposure, for detailed epidemiological studies in pig farms the examination of antibody isotypes will give more comprehensive information.     

Molecular characterization of Listeria monocytogenes and the significance for food hygiene

Research on the pathogen Listeria monocytogenes is a key issue both for the clinical and the food microbiologist owing to the unique pathway of infection and the exposure of humans via contaminated foods. Although, in Austria, the incidence of listeriosis is about 870-fold lower than the incidence for Salmonella infection, the food law manages both foodborne pathogens with a comparable stringency. The current risk management is based on the assumption that environmental L. monocytogenes isolates, from which the pool of "foodborne" isolates is recruited, are of similar pathogenicity compared to clinical and outbreak isolates. This verdict became doubted in the recent years. Characterization of L. monocytogenes by virulence gene sequencing, virulence studies in vivo and in vitro and by molecular typing was considerably stimulating the discussion on virulence variability in L. monocytogenes. This article provides insights in the value of epidemiological follow-up studies by presenting a typing study on 15 cases of listeriosis observed in a district hospital in Turkey. Furthermore results from typing L. monocytogenes either by virulence gene sequencing, mismatch amplification mutation assay or by pulsed field gel electrophoresis are discussed. The close interaction of molecular microbiology with food microbiology both in applied and basic science is currently creating a new discipline of molecular food microbiology. We are convinced that veterinary medicine will contribute to this exiting development in a fruitful way.     

Suitability of SSCP-PCR analysis for molecular detection of quinolone resistance in Campylobacter jejuni

Foodborne infections with Campylobacter spp. are increasing, especially antibiotic resistant strains are emerging. Quinolone resistant isolates can cause failure of therapy in severe clinical infections. Molecular characterisation is needed for the detection of resistant variants of C. jejuni. Therefore 23 isolates from poultry and human medicine as well as three control strains were tested for their minimal inhibitory concentration, their Single-Strand-Conformation-Polymorphism (SSCP)-PCR pattern (a method for the detection of resistance determining point mutations), and their sequence of the quinolone resistance determining region (QRDR). Six different SSCP types could be identified: two types for quinolone resistant isolates and other types containing so called silent mutations without influence on the resistance. A genotypic monitoring of the quinolone resistance in C. jejuni can be useful for the early detection of new resistance variants. As a screening method for detection of point mutations in the QRDR the SSCP-PCR can be applied. Compared to other genotypic methods the SSCP-PCR is less time and cost consuming and needs only standard technical equipment.     

Canine brucellosis--a case report

Brucella (B.) canis was isolated from ejaculate of a 4-year old Korthals-Griffon male dog after occurrence of epididymitis and orchitis. Despite several trials of therapy with different antibiotics relapes occurred, with B. canis being isolated from ejaculate, blood and urine samples, respectively. Bacteriological examinations were added by serological testing over a period of about 1.5 years. During the study SAT serum titre steadily dropped from 1:200 to 1:50. By CFT, B. canis antibodies were detectable at the beginning with a titre of 1:320 and to the end of the study with titres between 1:80 and 1:160.     

Effects of butyrate on apoptosis in the pig colon and its consequences for skatole formation and tissue accumulation

Evidence exists that butyrate inhibits apoptosis of colon crypt cells in vivo so that less tryptophan from cell debris is available for skatole formation by microbes in the pig colon. In this study, potato starch containing a high proportion of resistant starch was fed to test the hypothesis that increased butyrate formation will occur in the colon and contribute to reduced epithelial cell apoptosis, thus leading to reduced skatole formation and absorption. Two groups of six barrows were provided with catheters in the jugular vein and fed either a ration with pregelatinized starch (high ileal digestibility; controls) or potato starch (low ileal digestibility; PS) as the main carbohydrate. All pigs were fed 31 MJ of metabolizable energy and 381 g of crude protein per day. The controls were fed for 19 d. The PS group received the same control ration for 10 d, and then changed to the PS ration. The total feeding period of PS consisted of a 5 d adaptation period followed by another 19 d. In the continously sampled feces, pH, short chain fatty acids, and skatole were determined. Skatole was additionally measured in blood plasma that was sampled daily. After killing barrows at the end of the feeding period, fat tissue for skatole measurement and colon tissue for histological quantification of mitosis and apoptosis were obtained. Feeding potato starch led to a rapid 2.2 fold increase of fecal butyrate when compared both with the control period of the PS group and the control group (P < 0.001). PS feeding resulted in a decrease in pH from 7.3 to 5.3 (P < 0.001) and apoptosis from 2.06 cells/crypt to 0.90 cells (P < 0.01), whereas there was no change in mitosis. Consequently, skatole decreased both in feces (controls vs PS group: 120.0 vs 1.9 microg/g; P < 0.001) and in blood plasma (1.6 vs 0.2 ng/mL; P < 0.001). The mean concentration of skatole in fat tissue was 167 ng/g tissue in controls, and below the detection limit (0.8 ng/g) in the PS group (P < 0.001). It is concluded that butyrate-dependent inhibition of apoptosis in the colon due to potato starch feeding efficiently inhibits skatole production in barrows. Because of the depressed skatole levels, improved sensory quality of pork is possible.     

Effect of dietary fat sources on systemic and intrauterine synthesis of prostaglandins during early pregnancy in gilts

The present experiment was conducted to determine the influence of dietary fatty acids C18:2n-6 and C18:3n-3 on the modulation of intrauterine synthesis of prostaglandin E2 (PGE2) and F2alpha (PGF2alpha) during early pregnancy in pigs. Prostaglandin E2 in uterine fluid has been previously reported to be associated with embryo survival and development. Thirty-two Yorkshire-Landrace nulliparous gilts were randomly allocated to four diets containing 5% supplemental fat. The four dietary treatments were: HT, hydrogenated tallow (26.5% C16:0 and 54.8% C18:0); SO, sunflower oil (61.3% C18:2n-6); LO, linseed oil (50.4% C18:3n-3); and SO(CLA), a mixture of sunflower oil and conjugated linoleic acids to provide 20% CLA. Treatments started 2 d after the first pubertal estrus (d -21) and lasted for 36 d (slaughter), which was 15 d after the second estrus (d 0; insemination). Fatty acids and PGE2 were measured in the peripheral blood plasma on d -19, d -7, d 0, and d 14. Fatty acids in endometrial tissues and PGE2 and PGF2alpha in the uterine fluid collected on d 15 were also measured. Concentrations of fatty acids in the plasma reflected the content of fatty acids in the diet as early as d -7. From d -7, PGE2 concentrations in the plasma were higher in gilts fed SO compared with HT (P < 0.05). Plasma PGE2 concentrations were lower (P < 0.01) on d 14 in gilts fed LO compared with HT. Total PGF2alpha contents in the uterine fluid of gilts fed LO were more than 70% lower (P < 0.05) than for the HT group. A similar trend was observed for total PGE2 content and for the ratio PGF2alpha:PGE2, but the effect (LO vs HT) was less marked (P < 0.07 and P < 0.10, respectively). There was no effect of SO or SO(CLA) on total PGE2 contents in the uterine fluid. Dietary enrichment in C18:2n-6 and/or C18:3n-3 for early pregnant gilts can influence fatty acids in plasma and endometrial tissue and can modulate circulatory and intrauterine prostaglandins.     

Effects of probiotics: alone and in a mixture of Biosacc plus Zinc Bacitracin on the caecal microflora of Japanese quail

The aim of this study was to investigate the effects of two commercially available probiotics, alone and in combination with an antibiotic, on the caecal flora of Japanese quail (Coturnix coturnix japonica) reared under unstressed conditions. Thirty-four 90-day-old Japanese quail were selected for this study. The birds were divided into four groups, two groups of nine birds and two groups of eight birds. The animals in these groups were given feed containing 0.5 kg per tonne of probiotics alone (Protexin or Biosacc), a mixture of probiotic plus antibiotic (Biosacc plus Zinc Bacitracin at 0.5 and 0.0525 ppm, respectively), and a group where no supplement was added to be used as controls. The total count of aerobic bacteria, lactobacilli, enterobacteriaceae, coliforms, enteroccoci, salmonellae, sulphite-reducing anaerobic bacteria (clostridia), and pH values in the caecal content of the birds were examined. No significant differences were detected among the four groups for pH values and bacterial number (p>0.05), except for sulphite-reducing anaerobic bacteria (p<0.001). These results suggest that the use of probiotics alone and/or a mixture of a probiotic plus antibiotic as a feed supplement does not have a major suppressing effect on the majority of bacterial groups in the caecal flora of mature, healthy Japanese quail reared in unstressed conditions.     

Immunocytological confirmation of bone marrow metastases in a dog with cholangiocarcinoma

The case history of a four-year-old, male Bernese mountain dog is presented. Carcinoma cells were detected in the liver by ultrasound-guided fine-needle aspiration. Bone marrow aspirated from the iliac crest and the left femur showed a distinct infiltration by carcinoma cells. Immunocytological examination of the liver and bone marrow metastases showed a negative staining result for large spectrum cytokeratin (CK) KL1, a strong positive result for CK7 and a focal weak positive result for CK20. The dog was euthanased due to the grave prognosis. Histopathological examination revealed metastatic cholangiocarcinoma. The authors conclude that cytological and immunocytological examination of bone marrow aspirates should be used more frequently for the detection of distant metastases of carcinomas in small animal medicine.