Comparisons of sampling procedures and time of sampling for the detection of Salmonella in Danish infected chicken flocks raised in floor systems

Bacteriological follow-up samples were taken from 41 chicken (Gallus gallus) flocks in floor systems, where Salmonella enterica (Salmonella) had been detected either directly in bacteriological samples or indirectly by serological samples. Three types of follow-up samples were compared to each other within each flock: 1) 5 pairs of socks, analysed as 5 samples, 2) 2 pairs of socks, analysed as one sample, and 3) 60 faecal samples, analysed as one pooled sample. Agreement between sampling methods was evaluated by the following statistical tests: 'Kappa', 'The adjusted rand', McNemar's test for marginal symmetry, Proportion of agreement P0, P+, P-, and Odds Ratio. The highest agreement was found between the 2 types of sock sampling, while the lowest agreement was found by comparing 60 faecal samples with 5 pairs of socks. Two pairs of socks analysed as one pool appeared to be just as effective in detecting S. enterica as the 60 faecal samples. In broiler flocks, 5 pairs of socks were used both in the routine samples taken at about 3 weeks of age for the establishment of infection of the flock, and as one of the follow-up samples taken shortly before slaughter age, which means that the only notable differences between the 2 sampling rounds were the age of the broilers and of their litter. S. enterica was detected more frequently in samples from broilers about 3 weeks old, than in similar samples taken from broilers a few days prior to slaughter at ca. 33-40 days of age.     

Pharmacokinetics of an ampicillin/sulbactam (2:1) combination in rabbits

The pharmacokinetics of a 2:1 ampicillin-sulbactam combination in six rabbits, after intravenous and intramuscular injection at a single dosage of 20 mg/kg bodyweight (13.33 mg/kg of sodium ampicillin and 6.67 mg/kg of sodium sulbactam) were investigated by using a high performance liquid chromatographic method for determining plasma concentrations. The plasma concentration-time curves were analysed by compartmental pharmacokinetic and noncompartmental methods. The disposition curves for both drugs were best described by an open two-compartment model after intravenous administration and a one-compartment model with first order absorption after intramuscular administration. The apparent volumes of distribution calculated by the area method for ampicillin and sulbactam were 0.62 +/- 0.09 and 0.45 +/- 0.05 L/kg, respectively, and the total body clearances were 0.65 +/- 0.04 and 0.42 +/- 0.05 L/kg h, respectively. The elimination half-lives of ampicillin after intravenous and intramuscular administration were 0.64 +/- 0.11 and 0.63 +/- 0.16 h, respectively, whereas for sulbactam the half-lives were 0.74 +/- 0.12 and 0.77 +/- 0.17 h, respectively. The bioavailability after intramuscular injection was high and similar in both drugs (73.34 +/- 10.08% for ampicillin and 83.20 +/- 7.41% for sulbactam). The mean peak plasma concentrations of ampicillin and sulbactam were reached at similar times (0.20 +/- 0.09 and 0.34 +/- 0.15 h, respectively) and peak concentrations were also similar but nonproportional to the dose of both products administered (13.07 +/- 3.64 mg/L of ampicillin and 8.42 +/- 1.74 mg/L of sulbactam). Both drugs had similar pharmacokinetic behaviour after intramuscular administration in rabbits.     

The Use of Faecal Progestin Measurements to Monitor Reproductive Activity in Mashona Cows in a Smallholder Farming Area of Zimbabwe

The objectives of this study were to investigate the relationship between the concentrations of progesterone (P₄) in the peripheral circulation and progestins in the faeces of cows in a smallholder farming area and to monitor the ovarian activity of such cows. The study was carried out in Sanyati smallholder farming area, located 250 km southwest of Harare. Blood and faecal samples for P₄ and progestin determination, respectively, were obtained once a week from 35 Mashona type cows that belonged to farmers from three different villages. The cows were weighed once every month and all births were recorded. The concentrations of progestins in faeces and P₄ in plasma were positively correlated (r = 0.72, p<0.01). Most of the non-pregnant cows (65%) cycled during the rainy season. Most of those that were acyclic at this time were undergoing post-partum anoestrus. During the dry season, most cows (58%) became acyclic as forage became scarce and of poor quality. Thirty-two calves were born during the study period, mostly during the first or last two months of the year, when there was enough forage. The mean calving to first oestrus in 14 cows that recalved was 71±49 days. Eight of these 14 cows conceived again, the calving to conception interval being 173±94 days. The cows' liveweights were highest in April, at the end of the rainy season. By the end of the dry season, the cows had lost, on average, 15% of their peak weight.     

Detection of Oestrus in Bunaji Cows under Field Conditions

The objective of this study was to determine the efficacy of some aids for detectiong oestrus in 72 Bunaji cows synchronized using two injections of prostaglandin F₂ (PGF₂) 13 days apart. Blood samples to determine the accuracy of the aids and ovarian activity by radioimmunoassay technique were collected daily from the day of the second PGF₂ injection until oestrus or for 168 hours for `non-responders'. The aids for detecting oestrus, tail painting, KaMar detectors and a chin-ball mating device (CMD) were applied at the time of the second injection.The oestrus response rate was 73.6% and 61.1%, from the serum progesterone concentration and visual observation, respectively. Based on the total number of oestrus events observed, unaided visual observation, tail painting, KaMar and CMD detected 52.2%, 82.6%, 82.6% and 76.8%, respectively. Visual observation of standing oestrus alone failed to detect 47.8% of the occurrence of oestrus as shown by the concentration of progesterone in the serum and 30.4%, 30.4% and 24.6%, respectively, of the oestrus periods recorded by tail painting, KaMar and CMD. Twenty-eight cows showed abnormalities in progesterone concentration.These investigations showed that oestrus may be detected by using the aids in cows that have been regarded as `anoestrous' by visual observation of standing heat.     

Factors involved in the early pathogenesis of bovine Staphylococcus aureus mastitis with emphasis on bacterial adhesion and invasion. A review

Staphylococcus aureus is the most important and prevalent contagious mammary pathogen; it causes clinical and subclinical intramammary infection with serious economic loss and herd management problems in dairy cows. In vitro studies have shown that Staphylococcus aureus adheres to mammary epithelial cells and extracellular matrix components and invades into mammary epithelial as well as other mammary cells. Staphylococcus aureus strains from intramammary infection produce several cell surface-associated and extracellular secretory products. The exact pathogenic roles of most of the products and their effects on adhesion and invasion are not well evaluated. It is also known that mammary epithelial cell-associated molecules and extracellular matrix components interact with S. aureus during the pathogenesis of mastitis, but their roles on adhesion and invasion have not been characterized. The adhesion of S. aureus to epithelial cells may involve non-specific physicochemical interactions and/or specific interactions between bacterial cell-associated ligands and host cell surface receptors. In vitro adhesion depends on the S. aureus strain, the growth phase of the bacteria, the growth medium and the origin of the epithelial cells. Adhesion is hypothesized to be a prerequisite and crucial early step for mammary gland infection. Staphylococcus aureus invades mammary epithelial cells. It also invades other cells such as endothelial cells and fibroblasts. Bacteria are found enclosed in membrane bound vacuoles in the cytoplasm of mammary epithelial cells. Recent observations indicate that S. aureus escapes from the phagosome into the cytoplasm and induces apoptosis. The invasion into mammary epithelial cells may occur through an endocytic process that requires involvement of elements of the cytoskeleton or by direct binding of bacteria to epithelial cells through a process mediated by specific receptors that needs de novo protein synthesis by both cells. Thus, the recurrent subclinical infection may result from this intracellular existence of bacteria that are protected from host defenses and effects of antibiotics. This review emphasizes on recent findings on S. aureus adhesion to mammary epithelial cells and extracellular matrix components and invasion into mammary epithelial cells.     

Isolation and characterization of Mycoplasma arginini from camels (Camelus dromedarius) with pneumonia

Post-mortem examinations of 100 camels with pneumonic lesions were made at a local abattoir for Mycoplasma species. Sixteen isolates with indistinguishable biochemical and immunological characters were identified. The biochemical profile of these isolates showed that they were sensitive to digitonin, negative for urease production, glucose fermentation, and phosphatase activity but were positive for arginine hydrolysis. The identity of these isolates was further confirmed by disk growth inhibition test using a panel of specific antisera against selected reference Mycoplasma spp. Based on the biochemical profile and growth inhibition results, the camel isolates were identified as M. arginini. The pathological findings associated with M. arginini isolation consisted mostly of chronic interstitial pneumonia. The isolation rate of M. arginini from these specimens was 8.8%. These results suggest that the role of M. arginini in pneumonia in camels should be explored in greater detail.     

Neuroendocrine-immune interactions in fish: a role for interleukin-1

Bi-directional communication between the hypothalamus-pituitary-adrenal (HPA)-axis and the sympathetic nervous system with the immune system is crucial to ensure homeostasis. Shared use of ligands and especially receptors forms a key component of this bi-directional interaction. Glucocorticoids (GC), the major end products of the HPA-axis differentially modulate immune function. Cytokines, especially interleukin-1 (IL-1), tumour necrosis factor-alpha (TNF-alpha) and interleukin-6 (IL-6), ensure immune signalling to the neuroendocrine system. In addition, hormones from leukocyte origin such as corticotropin-releasing hormone (CRH), adrenocorticotropic hormone (ACTH) and beta-endorphin, as well as centrally synthesised and secreted cytokines, contribute to the communication network.In teleost fish cortisol is the major product of the hypothalamus-pituitary-interrenal (HPI)-axis which is the teleost equivalent of the HPA-axis. Moderate and substantial increases in cortisol during stressful circumstances negatively affect B-lymphocytes, whereas rescue of neutrophilic granulocytes may support innate immunity. Recent elucidation of lower vertebrate cytokine sequences has facilitated research into neuroendocrine-immune interactions in teleosts and the first evidence for a significant function of interleukin-1 in the bi-directional communication is discussed.     

Evaluation of a specific immunochemotherapy for the treatment of canine visceral leishmaniasis

The efficacy of specific immunochemotherapy against Leishmania infantum infection in dog was studied. The effects on transmission of the disease, as well as the cellular and humoral immune response were examined. The treated animals showed a significant reduction in the infection rates that were detected in Phlebotomus perniciosus females fed on the dog. The humoral immune response, assayed with an indirect immunofluorescence antibody test (IFAT), did not show significant variations under the influence of the therapy. The characterisation of the peripheral blood mononuclear cells (PBMC) using flow cytometry indicated a significant increase in the proportion of T lymphocytes, especially of CD4/TcR(alpha)(beta)(+) and CD4/CD45RA(+) cells, without showing evidence for modifications in the other leukocyte subsets. Cellular lymphoproliferation studies indicated a lack of a specific response to soluble leishmanial antigen (SLA), but the non-specific lymphoproliferative capacity assayed with phytohemagglutinin (PHA) was maintained.     

Isolation and characterization of immortalized porcine aortic endothelial cell lines

Primary porcine endothelial cells have a limited life span in culture. After four to five passages, they tend to de-differentiate and eventually reach senescence. The aim of this work was to establish immortalized porcine aortic endothelial cell lines (AOCs) to facilitate in vitro studies of different pathological process involving the endothelium. Primary porcine aortic endothelial cells (PAECs) were transfected with a plasmid containing the SV40 genome and selected on the basis of morphological and phenotypical features. Flow cytometry analysis demonstrated uptake of acetylated low density lipoproteins (Ac-LDL) and constitutive expression of SLA class I, CD29, CD31, CD41/61, CD80/86, CD46, SWC3, and LAMP-1 antigens by all analyzed lines and showed little differences to primary cells. The functional similarity between primary and immortalized endothelial cells was demonstrated in a cytotoxicity assay using a human natural killer cell line (NKL) as effector. The AOCs cell lines should be valuable tools for in vitro study of the human immune response against pig endothelial cells. In addition, they would be very useful to gain insight in the pathogenesis of some viral haemorrhagic diseases of pig such as African swine fever (ASF) or classical swine fever (CSF).