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Comparison of the omp I gene of Chlamydia psittaci between isolates in Victorian koalas and other animal species.

OBJECTIVE: The objective of this study is to compare the strain of chlamydia causing genital infection in koalas from Victoria with isolates from other animal species. DESIGN: Polymerase chain reaction and restriction enzyme analysis has been used to compare various Chlamydia psittaci isolates from a range of animals and disease syndromes. The isolates used in this study include isolates from three birds, three from aborted sheep, one from polyarthritis, one from bovine abortion, one from feline pneumonitis, three porcine isolates from faeces, polyarthritis and abortion, and three urogenital isolates from Victorian koalas. PROCEDURE: Two polymerase chain reactions were performed, each amplifying a different region of the omp 1 gene. The first polymerase chain reaction amplified a 144 bp segment of the gene which was then digested with the restriction enzyme EcoR I. The second polymerase chain reaction amplified a larger 1070 bp region of the omp 1 gene which was digested with two restriction enzymes Alu I and Nde II. RESULTS AND CONCLUSIONS: The results obtained have confirmed that variation in DNA sequence of various animal chlamydia isolates does occur. They have also shown that it is possible to classify isolates, based on their restriction enzyme profiles, into distinct groups.

Animals↗

Modelling the spread of foot-and-mouth disease in Australia.

Preparedness for an incursion of an exotic animal disease is of key importance to government, industry, producers and the Australian community. An important aspect of Australia's preparedness for a possible incursion of foot-and-mouth disease is investigation into the likely effectiveness and cost-efficiency of eradication strategies when applied to different regional outbreak scenarios. Disease modelling is a tool that can be used to study diseases such as foot-and-mouth disease to better understand potential disease spread and control under different conditions. The Australian Government Department of Agriculture, Fisheries and Forestry has been involved with epidemiologic simulation modelling for more than 10 years, and has developed a sophisticated spatial model for foot-and-mouth disease (AusSpread) that operates within a geographic information system framework. The model accommodates real farm boundary or point-location data, as well as synthesised data based on agricultural census and land use information. The model also allows for interactions between herds or flocks of different animal species and production type, and considers the role that such interactions are likely to play in the epidemiology of a regional outbreak of foot-and-mouth disease. The user can choose mitigations and eradication strategies from those that are currently described in Australia's veterinary emergency plan. The model also allows the user to evaluate the impact of constraints on the availability of resources for mitigations or eradication measures. Outputs include a range of maps and tabulated outbreak statistics describing the geographic extent of the outbreak and its duration, the numbers of affected, slaughtered, and, as relevant, vaccinated herds or flocks, and the cost of control and eradication. Cost-related outputs are based on budgets of the value of stock and the cost of mitigations, each of which can be varied by the user. These outputs are a valuable resource to assist with policy development and disease management.

Animals↗

Dietary vitamin D toxicity in a household of pot-bellied pigs (Sus scrofa).

Vitamin D intoxication developed in Vietnamese Pot-Bellied Pigs (Sus scrofa) fed 2 commercially available swine rations. Pronounced hypercalcemia and a history incompatible with other causes of hypercalcemia led to confirmation of this diagnosis by plasma vitamin D metabolite analysis in 2 affected animals as compared to a control animal. Feed sample analysis suggested the diet as the likely source of toxicity.

Animal Feed↗

Contributory and exacerbating roles of gaseous ammonia and organic dust in the etiology of atrophic rhinitis.

Pigs reared commercially indoors are exposed to air heavily contaminated with particulate and gaseous pollutants. Epidemiological surveys have shown an association between the levels of these pollutants and the severity of lesions associated with the upper respiratory tract disease of swine atrophic rhinitis. This study investigated the role of aerial pollutants in the etiology of atrophic rhinitis induced by Pasteurella multocida. Forty, 1-week-old Large White piglets were weaned and divided into eight groups designated A to H. The groups were housed in Rochester exposure chambers and continuously exposed to the following pollutants: ovalbumin (groups A and B), ammonia (groups C and D), ovalbumin plus ammonia (groups E and F), and unpolluted air (groups G and H). The concentrations of pollutants used were 20 mg m-3 total mass and 5 mg m-3 respirable mass for ovalbumin dust and 50 ppm for ammonia. One week after exposure commenced, the pigs in groups A, C, E, and G were infected with P. multocida type D by intranasal inoculation. After 4 weeks of exposure to pollutants, the pigs were killed and the extent of turbinate atrophy was assessed with a morphometric index (MI). Control pigs kept in clean air and not inoculated with P. multocida (group H) had normal turbinate morphology with a mean MI of 41.12% (standard deviation [SD], +/- 1. 59%). In contrast, exposure to pollutants in the absence of P. multocida (groups B, D, and F) induced mild turbinate atrophy with mean MIs of 49.65% (SD, +/-1.96%), 51.04% (SD, +/-2.06%), and 49.88% (SD, +/-3.51%), respectively. A similar level of atrophy was also evoked by inoculation with P. multocida in the absence of pollutants (group G), giving a mean MI of 50.77% (SD, +/-2.07%). However, when P. multocida inoculation was combined with pollutant exposure (groups A, C, and E) moderate to severe turbinate atrophy occurred with mean MIs of 64.93% (SD, +/-4.64%), 59.18% (SD, +/-2.79%), and 73.30% (SD, +/-3.19%), respectively. The severity of atrophy was greatest in pigs exposed simultaneously to dust and ammonia. At the end of the exposure period, higher numbers of P. multocida bacteria were isolated from the tonsils than from the nasal membrane, per gram of tissue. The severity of turbinate atrophy in inoculated pigs was proportional to the number of P. multocida bacteria isolated from tonsils (r2 = 0.909, P < 0.05) and nasal membrane (r2 = 0.628, P < 0.05). These findings indicate that aerial pollutants contribute to the severity of lesions associated with atrophic rhinitis by facilitating colonization of the pig's upper respiratory tract by P. multocida and also by directly evoking mild atrophy.

Air Pollution, Indoor↗

Morphological and antigenic relationships between viruses (rotaviruses) from acute gastroenteritis of children, calves, piglets, mice, and foals.

The reovirus-like particles present in the feces of young pigs and foals with acute enteritis and the virus causing epizootic diarrhea of infant mice were found to be indistinguishable morphologically from each other, from the South African SA. 11 and "O" viruses, and from the rotaviruses of children and calves. The inner capsid layer of each of these viruses reacted seriologically with sera of children, calves, mice, piglets, and foals convalescent from infection with their respective rotaviruses. These sera reacted by immunofluorescence with human, bovine, porcine, and murine rotaviruses, SA.11, and "O" viruses in tissue cultures and with human bovine, procine, nad murine viral antigens by complement fixation and gel diffusion. However, the antisera differed in their ability to react serologically with the outer capsid layer of the viruses investigated and in their ability to neutralize tissue culture-adapted calf virus. These two tests may demonstrate strain or host specificity among rotaviruses. Since the porcine, murine, and equine viruses are closely related serologically to and are morphologically identical to the human and bovine viruses, they should be included in the group of viruses for which the term "rotavirus" has been suggested. All known members of this proposed group of viruses share a common antigen, probably situated within the inner capsid layer; thus, any one of the viruses may be used for the preparation of antigen or antibody for diagnostic tests, and this will aid in the diagnosis of virus infection in those species from which a rotavirus has not been cultured.

Animals↗

Reproduction of porcine proliferative enteropathy with pure cultures of ileal symbiont intracellularis.

Porcine proliferative enteropathy is consistently associated with the presence of intracellular curved bacteria in epithelial cells in affected portions of intestine. Two strains of these intracellular bacteria were cultured in a cell culture system with rat enterocytes (IEC-18) and passaged several times and used as oral inocula for 14 gnotobiotic and 8 conventional pigs. DNA and immunological studies had identified these bacteria as belonging to a new taxon, Ileal symbiont (IS) intracellularis. Conventional pigs dosed with approximately 3.7 x 10(6) of these organisms passaged six times in cell culture developed severe lesions of proliferative enteropathy in the ileum. Other conventional pigs dosed with a lower titer or with organisms passaged 13 times developed moderate and minor lesions, respectively. All gnotobiotic pigs dosed with organisms failed to develop lesions. Control pigs, eight conventional and two gnotobiotic, dosed with diluent, uninfected cell material or left undosed failed to develop lesions also. Reisolation of IS intracellularis and demonstration of the organism in mucosal and fecal samples only occurred in conventional pigs dosed with organisms. Gnotobiotic pigs lacking a normal intestinal flora have not been shown to be colonized by the organism. Seroconversion to IS intracellularis or mucosal infiltration by inflammatory cells was not observed in experimentally affected pigs, confirming the weak immune response characteristic of the natural disease. These results support the identification of IS intracellularis as an etiological agent of proliferative enteropathy in pigs.

Animals↗

Occurrence of plasmids and antibiotic resistance among Campylobacter jejuni and Campylobacter coli isolated from healthy and diarrheic animals.

Serologically defined strains of Campylobacter jejuni and Campylobacter coli from healthy and diarrheic animals were examined for the occurrence of plasmid DNA in association with the antibiotic susceptibility of the bacterial host and the health status of the animal host. Of all campylobacter organisms surveyed, 53% (116 of 200) contained plasmid DNA. A plasmid occurrence rate of 73.8% was obtained for C. coli from healthy pigs, contrasted by lower plasmid occurrence rates for C. coli from diarrheic pigs (30%) and from all diarrheic animals (21.4%). For C. jejuni, in contrast, only 13.6% of healthy cattle contained plasmid DNA, contrasted by a higher plasmid occurrence rate of 31.2% from diarrheic cattle. A high plasmid occurrence rate of 75.8% was observed for C. jejuni from healthy chickens. Campylobacter plasmids ranged in size from less than or equal to 1 to 86 megadaltons. Antibiotic susceptibility for 52 animal isolates (excluding chickens) indicated that most isolates were susceptible to kanamycin, erythromycin, gentamicin, tetracycline, and compound sulfonamide, whereas few were susceptible to bacitracin (19.2%); approximately half were susceptible to ampicillin (55.8%) and streptomycin (51.9%), and no isolates were susceptible to penicillin G. More isolates containing plasmids were resistant to ampicillin, tetracycline, and gentamicin than were isolates not carrying plasmids, there being a statistically significant difference for tetracycline and gentamicin, which suggested that these two antibiotics were probably plasmid mediated. The antibiotic susceptibility patterns of 21 chicken isolates of C. jejuni, by contrast, were different in that most were susceptible to ampicillin in addition to kanamycin, erythromycin, and gentamicin, whereas few wer susceptible to compound sulfonamide, streptomycin, and tetracycline in addition to penicillin G and bacitracin. A 30- or 39-megadalton plasmid, or both, common to many of the chicken isolates was usually associated with tetracycline resistance.

Animals↗

Evaluation of the RapID NH system for identification of Haemophilus somnus, Pasteurella multocida, Pasteurella haemolytica, and Actinobacillus pleuropneumoniae isolated from cattle and pigs with respiratory disease.

Haemophilus somnus, Pasteurella haemolytica, Pasteurella multocida, and Actinobacillus pleuropneumoniae from cattle and pigs with respiratory disease were used to evaluate the RapID NH system (Innovative Diagnostics, Atlanta, Ga.). Minor modifications of the RapID NH system to include animal source and growth requirements would permit the identification of all isolates tested.

Actinobacillus pleuropneumoniae↗

Genetic characterization of type 2 porcine circovirus (PCV-2) from pigs with postweaning multisystemic wasting syndrome in different geographic regions of North America and development of a differential PCR-restriction fragment length polymorphism assay to detect and differentiate between infections with PCV-1 and PCV-2.

Postweaning multisystemic wasting syndrome (PMWS) is an emerging disease in swine. Increasing evidence indicates that a variant strain of porcine circovirus (PCV), designated type 2 PCV (PCV-2), is responsible for PMWS. To determine the extent of genetic heterogeneity of PCV-2 isolates, the complete genomes of six PCV-2 isolates from different regions of North America were amplified by PCR and sequenced. Sequence and phylogenetic analyses confirmed that two distinct genotypes of PCV exist: nonpathogenic genotype PCV-1 and PMWS-associated genotype PCV-2. However, within the PCV-2 genotype, several minor branches that have been identified appear to be associated with geographic origins. The genomic sequences of two French PCV-2 isolates diverge the most from those of other PCV-2 isolates and form a distinct branch. Other minor but distinguishable branches have also been identified for a Taiwan PCV-2 isolate and two of the Canadian PCV-2 isolates. All the U.S. PCV-2 isolates are closely related, but the Canadian isolates vary, to some extent, in their genomic sequences. The data from this study indicate that although the genome of PCV-2 is generally stable among different isolates, PCV-2 isolates from different geographic regions vary in their genomic sequences. This variation may have important implications for PCV-2 diagnosis and research. On the basis of genetic analyses of available PCV strains, a universal PCR-restriction fragment length polymorphism (PCR-RFLP) assay was developed to detect and differentiate between infections with PCV-1 and PCV-2. This PCR-RFLP assay should be useful for studying the pathogenesis of PCV-2, for detecting PCV-2 infection in pigs from different geographic regions, and for screening donor pigs for use in xenotransplantation.

Amino Acid Sequence↗

Development of a real-time PCR assay for detection of Toxoplasma gondii in pig and mouse tissues.

A highly sensitive and specific method has been developed to reproducibly detect and quantitate Toxoplasma gondii burden in animal tissue samples using T. gondii ITS1-derived primers and a fluorogenic probe via real-time PCR. Assay specificity was confirmed against a panel of DNA samples from T. gondii and other common protozoa as well as host animal tissue. This Toxo TaqMan assay was able to detect as little as 0.1 pg of T. gondii genomic DNA, which is equivalent to 1 T. gondii bradyzoite, and has a dynamic range of detection of from 100 ng to 100 fg of T. gondii DNA. Tissues from experimentally infected mice and pigs as well as bradyzoite-spiked pig muscle samples were used to test and standardize this technique. Positive signals were obtained with T. gondii parasite concentrations ranging from 4 to 3.7 x 10(5) parasites per g of spiked pig tissue, with excellent linearity (R(2) = 0.9776). All T. gondii-infected animals were correctly identified by this technique. Results indicate that this assay is applicable to swine carcasses and commercial pig products, is compatible with automation technology for potential slaughterhouse use, and will enable scientists to diagnose and quantitate T. gondii in animal tissues.

Animals↗

Identification and quantification of ovine gammaherpesvirus 2 DNA in fresh and stored tissues of pigs with symptoms of porcine malignant catarrhal fever.

Cases of porcine malignant catarrhal fever were analyzed by a combination of identification and quantitation of ovine gammaherpesvirus 2 DNA in a variety of paraffin-embedded tissues from diseased pigs, serology, and exclusion of primary porcine gammaherpesviruses. In spite of reduced signal due to fixation and paraffin embedding, ovine gammaherpesvirus 2 DNA in pig brains exceeded the amounts found in sheep brains by orders of magnitude.

Animals↗

Characterization of vancomycin-resistant Enterococcus faecium isolates from broiler poultry and pig farms in England and Wales.

This study aimed to investigate the occurrence and molecular epidemiology of vancomycin-resistant Enterococcus faecium (VREF) isolates on poultry and pig farms in England and Wales. A total of 217 VREF isolates were obtained from fresh feces and environmental swabs collected from conventional and organic farms. A predominant pulsed-field gel electrophoresis (PFGE) profile was found for each VREF-positive farm, together with less frequent types. All isolates presented the vanA genotype and were esp negative. Seventy-six percent of the VREF isolates were additionally resistant to nine or more antimicrobials, presenting a diverse range of resistance phenotypes. The multiresistance traits did not appear to be specific to individual farms or sample types (i.e., environmental or fecal), nor did they correlate with any specific PFGE type. Ninety-three percent of the isolates were resistant to penicillin, 89% were resistant to tetracycline, 87.5% were resistant to erythromycin, and 50% were resistant to quinupristin-dalfospristin (Synercid). The lack of clonality among these populations may suggest the horizontal transfer of resistance genes and/or a dynamic replacement of clonal lines rather than persistence.

Animal Husbandry↗

"ONO" syndrome.

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Animals↗