Adherence of Actinobacillus pleuropneumoniae to primary cultures of porcine lung epithelial cells

To study adherence of Actinobacillus pleuropneumoniae to porcine lower respiratory epithelium, a cell culture model was developed using primary cultures of porcine lung epithelial cells (LEC). Adherence assays were performed and results were compared with data obtained with swine kidney cells (SK6). A. pleuropneumoniae efficiently adhered to LEC with up to 62 bacteria per cell after 2h of incubation. Reference strain of serotype 3 (R3) adhered better to LEC than reference strains of serotypes 1 (R1), 7 (R7) and 8 (R8). Overall the adherence to LEC was more rapid and up to 30-fold more efficient than adherence to SK6 cells. In search for the mechanism involved in the adherence event, we tested the effect of LPS which has previously been demonstrated to cause adherence of the pathogen to upper respiratory epithelium. Adherence assays with LPS transposon mutants demonstrated unaltered (mutant with modification in core/lipid A moiety) or even three-fold more adherence (mutants lacking O antigen) compared to the parent micro-organisms. Purified LPS of strains R1, R3, R7 and R8 did not inhibit adherence of R8 to LEC either, suggesting that LPS and particularly the O-antigen are not essential for adherence of A. pleuropneumoniae to LEC. The efficient, LPS-independent adherence of A. pleuropneumoniae to LEC cells indicates that A. pleuropneumoniae may carry different, cell type-specific adhesins and that primary cultures of lower respiratory epithelium are valuable infection models in studying A. pleuropneumoniae pathogenesis.     

Risk factors associated with behavioural disorders of crib-biting,weaving and box-walking in Swiss horses

REASONS FOR PERFORMING STUDY: Studies on the prevalence of behavioural disorders in horses and on associated risk factors have revealed inconsistent results. There are many studies on the neuropharmacological, surgical or mechanical therapy of stereotypies, but little is known about their causation. OBJECTIVES: To explore risk factors associated with the occurrence of behavioural disorders in horses. METHODS: A sample of horse owners, selected randomly and representative for Switzerland, was contacted in a postal survey. Answers were provided for 622 stables (response rate 35.2%). Individual data of 2,341 horses were examined with path analysis (multivariable linear and logistic regression), and adjustment made for possible confounding effects due to age and breed. RESULTS: Out of 60 possible risk factors, 11 were associated with the outcome at the univariable level (null-hypothesis path model) and 3 factors remained after the backward logistic regression procedure. Mature Warmbloods and Thoroughbreds, assessed by the owners to be reactive, fed 4 times a day and without daily pasture, had increased odds of displaying crib-biting, weaving and box-walking. Furthermore, indirect associations of 5 factors with the outcome were identified. CONCLUSIONS: The final logistic regression model of risk factors leads to the hypotheses that causal prevention of stereotypic behaviours should be based upon housing and management conditions which allow tactile contact with other horses (e.g. mutual grooming), daily free movement (paddock or pasture), as well as the provision of high amounts of roughage but of little or no concentrates. POTENTIAL CLINICAL RELEVANCE: It is one of the aims of population medicine to prevent the development of behavioural disorders. Further research is needed to test the concluding hypotheses in experimental studies or to verify them in the context of similar observational studies.     

Virulence factors of avian pathogenic Escherichia coli isolated from broilers from the south of Brazil

Sixty-three Escherichia coli strains isolated from broilers with respiratory problems were examined for virulence factors, hemolysin synthesis ability, motility, hemagglutination capacity, operon pap presence, colicin production, and serum resistance. The capacity to hemagglutinate guinea pig erythrocytes was found in 53 (84.1%) of the samples, but only 30 (47.6%) agglutinated chicken erythrocytes. D-mannose-sensitive hemagglutination against guinea pig erythrocytes was found in 19 (30.2%) samples and against chicken erythrocytes, in 15 (23.8%) samples, whereas the D-mannose-resistant hemagglutination with guinea pig erythrocytes was found in 34 (54%) samples, and 13 of these (20.6%) showed this characteristic against chicken erythrocytes. Operon pap, P fimbria codifier, was detected in 26 samples in a total of 34 D-mannose-resistant samples. Colicin production was observed in 55 (87.3%) of the strains, and 41.8% presented V colicin production. Of the samples analyzed, 56 (88.9%) presented serum resistance, six (9.5%) were intermediate, and only one (1.6%) was sensitive to the action of the complement. The diversity of virulence profiles detected in the samples in this study explains in part the multifactorial characteristics of avian colibacillosis.     

Facet joint geometry and intervertebral disk degeneration in the L5-S1 region of the vertebral column in German Shepherd dogs

OBJECTIVE: To evaluate the possible association between facet joint geometry and intervertebral disk degeneration in German Shepherd Dogs. ANIMALS: 25 German Shepherd Dogs and 11 control dogs of similar body weight and condition. PROCEDURE: Facet joint angles in the caudal portion of the lumbar region of the vertebral column (L5-S1) were measured by use of computed tomography, and the intervertebral discs were evaluated microscopically. The relationship between facet joint geometry and disk degeneration was evaluated by use of statistical methods. RESULTS: German Shepherd Dogs had significantly more facet joint tropism than control dogs, but an association with disk degeneration was not found. However, German Shepherd Dogs had a different facet joint conformation, with more sagittally oriented facet joints at L5-L6 and L6-L7 and a larger angle difference between the lumbar and lumbosacral facet joints, compared with control dogs. CONCLUSIONS AND CLINICAL RELEVANCE: A large difference between facet joint angles at L6-L7 and L7-S1 in German Shepherd Dogs may be associated with the frequent occurrence of lumbosacral disk degeneration in this breed.     

Oral administration of yeast, Saccharomyces cerevisiae, enhances the cellular innate immune response of gilthead seabream (Sparus aurata L.)

The effects of including lyophilised whole yeast, Saccharomyces cerevisiae, in the diet on the seabream innate immune response were investigated. Gilthead seabream (Sparus aurata L.) specimens were fed four different diets for 4 weeks: a commercial diet as control and the same diet supplemented with 1, 5 or 10 g/kg yeast. After 1, 2 and 4 weeks, serum complement titres, as a humoral parameter, and phagocytic, respiratory burst, myeloperoxidase and natural cytotoxic activities of head-kidney leucocytes, as cellular parameters, were evaluated. The results showed that yeast supplements enhanced all the latter responses, but not the humoral response. This enhancement was dose-dependent except for the cytotoxic activity that was only stimulated by the lower dose of yeast assayed. As yeast cell walls are able to enhance the seabream cellular innate immune response, these results support the possible use of whole yeast as natural inmunostimulants in common fish diets.     

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).     

Three-dimensional kinematics of the equine interphalangeal joints: articular impact of asymmetric bearing

The objective of this study was to assess the effects of asymmetric placement of the foot on the three-dimensional motions of the interphalangeal joints. Four isolated forelimbs were used. Trihedrons, made of three axes fitted with reflective markers, were screwed into each phalanx. They allowed to establish a local frame associated with each bone and thus to define the spatial orientation of the phalanges. The limbs were then placed under a power press, and subjected to compression with gradually increasing force (from 500 to 6 000 N). The procedure was performed in neutral position and with the lateral or medial part of the foot raised by a 12 degrees wedge. Flexion, collateromotion (passive abduction/adduction) and axial rotation of the interphalangeal joints were measured using a cardan angle decomposition according to the principle of the "Joint Coordinate System" described by Grood and Suntay. Raising the lateral or medial part of the hoof induced collateromotion (about 5.6 degrees +/- 0.8) and axial rotation (about 6.5 degrees +/- 0.5) of the distal interphalangeal joint. The proximal interphalangeal joint underwent axial rotation (about 4.7 degrees +/- 0.5 at 6 000 N) and slight collateromotion. Both interphalangeal joints underwent collateromotion in the direction of the raised part of the foot (i.e., narrowing of the articular space on the side of the wedge), whereas axial rotation occurred in the direction opposite to the raised part of the foot. These results confirm the functional importance of interphalangeal joint movements outside the sagittal plane. In particular they demonstrate the involvement of the proximal interphalangeal joint in the digital balance. These data are helpful for the identification of biomechanical factors that may predispose to interphalangeal joint injury. Also the data may be of use for the rational decision making with respect to exercise management and corrective shoeing of the lame horse.     

Effect of dietary conjugated linoleic acids on the distribution of fatty acids in serum lipoprotein fractions and different tissues of growing pigs

The effect of conjugated linoleic acid (CLA) on blood lipids [total cholesterol, high-density lipoprotein (HDL), low-density lipoprotein (LDL) and triacylglycerols (TAG)] and the fatty acid distribution of the lipoprotein fractions, backfat, muscle fat, and liver lipids were examined in an experiment with two groups of 40 pigs [Pietrain x (Landrace x Large White)] each. The 20 female and 20 male castrated pigs of each group were fed with isoenergetic and isonitrogenous diets supplemented with either 20 g/kg rapeseed oil (control) or 20 g/kg CLA-TAG. The CLA preparation contained 54.2% pure CLA consisting of approximately two-thirds cis,trans/trans,cis-isomers and one-third trans,trans-isomers. The fatty acids of lipoproteins, backfat, muscle lipids and liver lipids were analysed by gas chromatograph (GC). CLA supplementation did not significantly influence blood lipids and the LDL to HDL ratio. In the CLA-fed pigs the very low-density lipoprotein (VLDL) contained higher saturated fatty acid (SFA) concentrations at the cost of the monounsaturated fatty acids (MUFA). The percentage of polyunsaturated fatty acids (PUFA) remained unchanged. The highest CLA content was analysed in VLDL (4.00%) followed by LDL (2.78%) and HDL (1.45%). The ratio of cis,cis to trans,trans isomers increased from VLDL over LDL to HDL. The content of SFA, probably in backfat and muscle lipids, increased whereas the part of MUFA decreased as a result of reduced Delta9-desaturase activity. The percentage of PUFA (without CLA) was higher in backfat of the control group in accordance with the dietary PUFA supply. This shift in the fatty acid distribution was not observed in the liver lipids. In all the three tissues analysed, the CLA-fed pigs had a significantly increased CLA content: the highest increase was in the backfat (5.65%), followed by liver lipids (2.41%), and muscle lipids (1.47%). An isomer-specific accumulation was observed for cis,cis-CLA isomers in muscle, and for trans,trans-CLA isomers in backfat. We conclude that CLA supplementation results in a higher SFA content in backfat and muscle lipids but not in liver lipids. There is a discrimination of the trans-10, cis-12 isomer and the trans,trans isomers in the formation of the cell membranes.     

Delayed acute capture myopathy in three roe deer

Delayed acute capture myopathy is the term used to describe the clinical syndrome observed in three roe deer captured by drive-nets and transported to an enclosure for scientific purposes. The animals died 48 h, 60 h and 8 days after being captured. The simultaneous deaths coincided with a previous episode of deliberate human disturbance. The histopathological findings were indicative of acute myopathy and myoglobinaemic nephrosis. These could be related to an ataxic myoglobinuric syndrome brought on by capture and transport operations. The lack of clinical signs and negative prognosis indicators in the period between capture and just before death. the absence of gross muscular lesions in the animal that died after 8 days post-capture, the simultaneous deaths of animals captured at different times and the evidence of deliberate human disturbance in the enclosure are suggestive of death triggered by a second stress episode.