Detection of Mycobacterium avium ssp. paratuberculosis in ileocaecal lymph nodes collected from elderly slaughter cows using a semi-nested IS900 polymerase chain reaction

The aim of this study was to investigate the occurrence of subclinical Mycobacterium avium spp. paratuberculosis (MAP) infections at slaughter by testing ileocaecal lymph nodes with a semi-nested IS900 PCR. Tissue samples were available within the framework of a parallel study investigating BSE-susceptibility factors in members of BSE-cohorts in the German Federal State of Lower Saxony. Ileocaecal lymph nodes were collected over a 2-year sampling period from 99 slaughter cattle of a mean age of 6.5 years (5.5-7.5 years). A recently developed IS900 semi-nested polymerase chain reaction (snPCR) assay offering a sensitivity of 1 genome equivalent was used for the detection of MAP-DNA. Based on this snPCR, 17 out of the 99 samples gave positive results, indicating a MAP occurrence of 17.17% in the random sample. All PCR products were sequenced for screening of polymorphisms. Nucleotide homologies of 98.5-100% were found with respect to the MAP K10 reference sequence IS900 (GenBank: AE16958). PCR analysis of ileocaecal lymph nodes collected from slaughter cattle proved to be a suitable technique to determine MAP occurrence in the local cattle population.     

Procollagen type-III aminoterminal peptide in serum and synovial fluid of dogs with hip dysplasia and coxarthrosis

Hip dysplasia is an affection of the coxofemoral joint that progresses until stabilization is caused by fibrosis and osteoarthritic changes. This stabilization process can be examined by clinical and radiographic methods. The capability of evaluating the procollagen concentrations in liquids, such as serum and synovial fluid, has further offered the basis for an objective biochemical evaluation of the stabilization process. Our study was performed to evaluate whether determination of procollagen concentrations was suitable for the use in practice. The procollagen type-III aminoterminal peptide (P-III-NP) concentration was measured in serum and in synovial fluid from coxofemoral joints in 20 dogs. Dogs were grouped on the basis of evidence of dysplasia and osteoarthritic changes of the hip: (1) a control group of 6 dogs without clinical or radiographic signs of hip dysplasia, and (2) dysplastic group of 14 dogs, which was further grouped with respect to the coxofemoral joint laxity, as determined by the Ortolani test. Synovial fluid concentration of P-III-NP was significantly (P less than 0.05) higher in fluid from dysplastic joints than in fluid from normal joints. Serum concentrations of P-III-NP were significantly (P less than 0.05) higher in dogs in which results of the Ortolani test were positive.     

A multivariable prognostic model for equine colic patients

A survey of 1965 equine colic cases was conducted from August 1985 to July 1986 ten equine referral hospitals located througout the U.S.A. Two-thirds of the cases were randomly selected for model development (1336), while the remaining cases (629) were used only for subsequent validation of the model. The following outcomes were defined: (1) died or killed prior to discharge from the hospital; or (2) alive at the time of discharge. Only variables which were significant (P<0.05) in an initial bivariate screening procedure and for which there were less than 400 missing values were considered in the multivariable modelling. A multivariable logistic regression model was constructed using a stepwise algorithm. The model used 666 cases and included the following variables: peripheral pulse (normal or weak), pulse rate, surgical or medical treatment, packed cell volume, self-inflicted trauma (absent or present) and capillary refill time. For each horse in the validation data set which had the appropriate variables recorded (n=335), the estimated expected probability of death (expected value) was calculated using the logistic regression equation. Using Bayes theorem, the post-test probability was calculated from the expected value (an estimate of the test odds) and the present probability (the case-fatality rate at each institution). Nomograms of predictive values for different case-fatality rates and expected values were constructed. Hosmer-Lemeshow goodness-of-fit statistics indicated that the model fitted the model data set well but the validation set poorly. However, when the observed case-fatality rates were compared with the average post-test probabilities for 0.10 increments of post-test probability, qualitatively, the model's performance was better.     

Horses' behavior and heart rate in a preference test for shorter and longer riding bouts

A Y-maze preference test was used to investigate whether horses prefer a shorter riding treatment over a longer riding treatment. In a pilot study (n = 4 horses), the maze was positioned with the 2 arms each pointing toward one of the short sides of the indoor arena, and in the main study (n = 14), the maze was rotated by 90°. Horses were 11 ± 4.4 years old and ridden regularly for at least 5 times/wk. They were conditioned to associate 1 exit of the maze with 1 lap of riding (R1), covering a distance of approximately 40 m at the walk or trot according to a predetermined schedule, and the other exit with 2 such laps (R2). Immediately afterward, riders dismounted, horses were led into the maze, and horses were let loose to make their choice in the maze. After exiting the maze, the rider mounted again and rode according to the chosen treatment. This procedure was repeated on the same day until statistical significance (P < 0.05) of preference was reached or up to a maximum of 35 trials. In addition, behavioral observations and heart rates were recorded. In the pilot study, all horses, regardless of the associated treatment, chose the left arm, which, unfortunately, pointed toward the arena's exit door. If horses were not caught immediately after exiting the maze (n = 5 occasions), they walked or trotted straight to the door. In the main study, 4 horses significantly preferred R1, 2 horses significantly preferred R2, and 8 horses had no significant preference. Heart rates were significantly (P < 0.05) higher during R2 (87.4 ± 2.6 bpm) than during R1 (79.5 ± 2.4 bpm). Except for tail swishing, no significant differences were found for the frequency of occurrence of behavior patterns between R1 and R2. Over the course of repeated trials, some horses became increasingly reluctant to enter and walk through the maze, and most showed increasing resistance to being remounted (e.g., sidestepping). Overall, the experimental setup did not seem to be appropriate to answer the research question. It is likely that the repeated mounting and dismounting caused discomfort or confused many of the horses to an extent that they did not actively select a treatment but rather searched for ways to evade further mounting (and riding). In conclusion, horses did not show a clear preference for either shorter or longer riding bouts, but their behavioral reactions indicate that they perceived mounting as uncomfortable and that their motivation to rejoin their herd-mates and/or to obtain feed in the barn was greater than their motivation to being ridden at all. Nevertheless, pronounced individual differences also seem to exist, with some horses showing little aversion to, and perhaps enjoyment of, being ridden, whereas others clearly preferring not to be ridden.     

Generation of Salmonella ghost cells expressing fimbrial antigens of enterotoxigenic Escherichia coli and evaluation of their antigenicity in a murine model

Salmonella Typhimurium ghost cells expressing K88ab, K88ac, K99, and FasA fimbriae of enterotoxigenic Escherichia coli (ETEC) in their envelopes were constructed. The genes encoding the fimbriae were individually cloned into an expression plasmid, pMMP81, carrying the asd gene, which was subsequently electroporated into the Δasd S. Typhimurium mutant. Plasmid pJHLP99, carrying the phiX174 lysis gene E, was also subsequently electroporated into the Salmonella mutant. The presence of the individual fimbriae on the ghost cells was examined by Western blot analysis. Forty BALB/c mice were equally divided into 2 groups of 20 mice each. Group A mice were intramuscularly vaccinated with a mixture of the 4 ghost cells expressing the individual fimbriae. The group B mice were inoculated with sterile phosphate-buffered saline as a control. The antigen-specific serum IgG concentrations were significantly higher in group A than in group B from week 2 until week 6 after inoculation. In addition, the antigen-specific IgA concentrations in fecal samples were significantly higher in group A than in group B at week 2 after inoculation. A large difference between the groups in the number of antigen-specific IgA-secreting cells in the small intestine was observed by immunohistochemical study. Also, the splenic lymphocyte proliferative responses were significantly greater in group A than in the control mice. These results suggest that vaccination with our Salmonella ghost cells can induce both humoral and cell-mediated immune responses and that the increased number of antigen-specific IgA-secreting cells in the small intestine may be correlated with the elevated fecal IgA immune response.     

Evaluation of an air-filtration system for preventing aerosol transmission of Porcine reproductive and respiratory syndrome virus

The purpose of this study was to evaluate the ability of a commercial air-filtration system to reduce aerosol transmission of Porcine reproductive and respiratory syndrome virus (PRRSV). The system consisted of a pre-filter and 2 filters with EU8 and EU13 ratings. In each of 4 trials, 5 PRRSV-infected donor pigs and 1 naive recipient pig (each 25 kg) were housed in opposing chambers connected by a 1.3-m-long duct. The system filtered air entering 1 recipient-pig chamber (filtered facility) from the donor-pig chamber but not a 2nd recipient-pig chamber (nonfiltered facility). The donor pigs had been experimentally infected with PRRSV MN-184, an isolate previously documented to be shed at a high frequency in contagious aerosols. On days 3 to 7 after infection of the donors, the 2 groups were housed in their respective chambers for 6 h and then in separate facilities, where samples were collected for testing by polymerase chain reaction and enzyme-linked immunosorbent assay over 14 d. Aerosol transmission was observed in 6 of the 20 replicates in the nonfiltered facility, whereas all pigs remained PRRSV-negative in the filtered facility; the difference was significant at P < 0.01. Thus, under the conditions of this study, the air-filtration system evaluated appeared to be highly effective at reducing aerosol transmission of PRRSV.     

Hemostatic changes in dogs with naturally occurring sepsis

Sepsis is a frequent source of morbidity and mortality in critically ill patients. The goal of this case control study was to measure hemostatic changes in dogs with naturally occurring sepsis. Blood was collected within 24 hours of admission from 20 dogs that fulfilled the criteria for sepsis. Sepsis was defined as histologic or microbiological confirmation of infection and 2 or more of the following criteria: hypo- or hyperthermia, tachycardia, tachypnea, or leukopenia, leukocytosis, or > 3% bands. Culture and sensitivities were performed on appropriate samples from all septic dogs. Twenty-eight control dogs were enrolled on the basis of normal results of physical examination, CBC, serum biochemistry, and coagulation profile. Plasma samples were analyzed for prothrombin time (PT), partial thromboplastin time (PTT), fibrin(ogen) degradation products (FDP), D-dimer (DD) concentrations, antithrombin (AT) activity, and protein C (PC) activity. Data were compared between groups by chi-square or independent t-tests. PC (P < .001) and AT (P < .001) activities were significantly lower in dogs with sepsis compared to controls. Dogs with sepsis had significantly higher PT (P = .007), PTT (P = .005), D-dimer (P = .005), and FDP (P = .001) compared to controls. Platelet counts were not significantly different between groups. Ten of the 20 septic dogs (50%) died, but no association was identified between any of the measured variables and outcome. These findings are consistent with previous studies in animals with experimentally induced disease and in clinical studies of humans. On the basis of these results, further investigation of the role of AT and PC in canine sepsis is warranted.