Corynebacterium kutscheri infection in wild voles (Microtus agrestis).
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Biomedical subjects
Publications and source records attributed to P A Barrow.
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The prevalence of infections caused by Mycobacterium bovis was estimated in two populations of wild animals in the south west of England. A variety of mammalian species was trapped on farm land where incidents of infection with tuberculosis had occurred in cattle. Bacteriological analysis of lymph node samples and lesions showed that the only animals acting as a reservoir for M. bovis were badgers. Examination of arthropod ectoparasites of infected badgers proved negative for M. bovis and suggests that these are unlikely to act as vectors in transmission of the disease.
A relatively simple method for the detection of intra-specific variation of Mycobacterium bovis is described, based on the ability of strains to utilize or remove amino acids from solution. Using the method, 31 strains of M. bovis, isolated badgers and cattle, have been divided in to four major groups with some additional sub-groups and intermediate strains. Preliminary results suggest that the biotypes observed are relatively stable. The method was used in the investigation of an outbreak of bovine tuberculosis in cattle in Gloucestershire where badgers were implicated as a source of infection. The results suggested that transmission of infection had occurred on at least two or three separate occasions. It is suggested that the method could be used in studying the epidemiology of bovine tuberculosis in wild animals, cattle and man.
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When pigs are weaned at two days of age large numbers of Excherichia coli appear in the anterior gut and the incidence of diarrhoea rises. The two phenomena do not appear to be directly related because the strains of E coli isolated are not serotypes previously found to be associated with neonatal pig scouring. Representative strains of the non-enteropathogenic serotypes did not produce enterotoxin and did not adhere to small intestine brush borders. Moreover when antibiotics were fed to eliminate E coli from the gut, the pigs still scoured. Rotavirus was detected in the gut contents and gut epithelium of scouring pigs and a bacteria-free filtrate of gut contents produced diarrhoea when administered to germ-free pigs. It is suggested that rotavirus may be one of the causes of the scouring seen shortly after weaning pigs at two days of age.
Light and electron microscopy showed lactobacilli and, to a lesser degree, streptococci to be closely associated with the squamous area of the pig stomach known as the pars esophagea. Several different types of extracellular layers were seen on bacteria attached to the epithelial surface. The total number of bacteria per square centimeter did not change with age up to 10 days, and there was no effect of weaning at 2 days. Lactobacillus fermentum, L. salivarius, and Streptococcus salivarius were isolated more frequently from sucking pigs than from those that were early weaned, whereas the reverse was true of L. acidophilus and S. bovis. All isolates recovered from washed macerated pars esophagea adhered to pig esophageal epithelial cells when tested in vitro.
The gastrointestinal microflora and gastric physiology of piglets weaned at 2 days was compared with that of piglets allowed to continue sucking the sow. Although there was a significantly higher count of Escherichia coli in the stomach, duodenum, and jejunum of the early-weaned compared with sow-reared pigs, these differences were not detectable in samples from the ileum. There were no quantitative differences in lactobacilli and in streptococci between the two treatments. Lactobacillus fermentum, L. acidophilus, Streptococcus salivarius, S. bovis, and related biotypes were isolated from both groups of pigs. L. fermentum and S. salivarius were isolated more frequently from sow-reared piglets. The weight of digesta in the stomach was greater in weaned than in sucking pigs and was even greater in scouring weaned pigs, suggesting that in scouring pigs there may be gastric stasis. The gastric pH was higher in the weaned pigs at 4 days of age, but gradually decreased up to 10 days, during which time the lactic acid concentration rose. In weaned pigs there was a highly significant negative correlation between pH and lactic acid concentration in the stomach digesta, and also a positive correlation between pH and number of E. coli. These correlations suggest that lactic acid, from bacterial fermentation, is the major component in the regulation of gastric pH in weaned pigs. Three of twenty sucking pigs, but none of the weaned pigs, were secreting HCl (chloride concentration > 3 mg/g, pH < 3.5). In sucking pigs there was an inverse relationship between the chloride and lactic acid concentrations in the digesta. In weaned scouring pigs there was a nonsignificant increase in pepsin concentration in the stomach tissue. There was a threefold increase in the total proteolytic activity of the stomach tissue.
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Salmonellosis remains an important human and animal problem worldwide and, despite extensive research effort, many of the details of its pathogenesis are not known. While there have been recent advances in some aspects of pathogenesis, other areas are not understood. The host adaptation shown by several serotypes and the recent dramatic changes in the predominance of particular serotypes are examples. Molecular techniques using in vitro model systems have identified several genes involved in adhesion and invasion, though their function and even their relevance to disease remain poorly defined. Similarly, several potential toxins have been identified and the genes cloned, although their significance is far from clear. Some of the essential genes on the large virulence plasmids have been defined, and these are known to be necessary for the establishment of systemic infection. Two of these genes are regulatory, but the function of the other genes is unknown. A general theme has been the identification of gene systems involved in regulation of virulence. New vaccines, based on 'rational attenuation' are being designed, and these have also been used to carry heterologous antigens; such vaccines are currently undergoing trials. The improved understanding of the pathogenesis of salmonellosis may also provide a model of wide applicability to a more general understanding of bacterial pathogenesis. New techniques, including the polymerase chain reaction, are being applied to diagnose salmonellosis.
A nuoG mutation in NADH dehydrogenase I was introduced into a virulent strain of Salmonella serotype Gallinarum, the causative agent of fowl typhoid, using gene replacement with a nuoG open reading frame inactivated by insertion of DNA encoding a kanamycin resistance determinant. The S. Gallinarum nuoG mutant, named SG9NGK, was highly attenuated in chickens. SG9NGK colonized the caeca of chickens less efficiently than the S. Gallinarum parental strain, was less invasive and showed no evidence of multiplication in the liver or spleen. Using a single oral immunization with live bacteria SG9NGK reduced mortality in 2-week-old chickens following challenge with virulent S. Gallinarum from 75% to less than 8%.
The serological response to Salmonella pullorum and S. gallinarum infection in chickens was studied with an indirect enzyme-linked immunosorbent assay (ELISA). In broiler chickens, a more virulent strain of S. pullorum produced a significantly lower serum IgG titer than did a less virulent strain. In laying hens, the serum and egg-yolk IgG titers were very similar. In chickens infected with S. gallinarum, high IgG titers persisted for 30 weeks. In chickens reinfected with this strain, each reinfection was followed by transitory increases in IgG lasting no longer than 2 weeks. Serum samples from Brazil taken from a laying flock with evidence of fowl typhoid showed much higher antibody levels than did those from three uninfected flocks. Using lipopolysaccharide as the detecting antigen, infections caused by these salmonellae could be differentiated from those caused by other groups. Incorporation of the appropriate flagella antigen in the ELISA allowed differentiation between infections caused by S. pullorum and S. enteritidis.
Four-day-old chickens infected orally with a spectinomycin-resistant (Spcr) mutant of a highly invasive avian Salmonella typhimurium strain excreted salmonellae in the feces for at least 10 weeks. When these chickens were reinfected at this time with a nalidixic acid-resistant (Nalr) mutant of the same strain, they excreted this mutant in significantly smaller numbers (P less than 0.01) than did a previously uninfected control group. The Nalr mutant had a shorter survival rate in the tissues of the immunized chickens than in tissues of the control birds. The Spcr mutant stimulated strong IgG, IgA, and IgM responses in serum, small-intestinal contents, and bile. These were detected by enzyme-linked immunosorbent assay (ELISA) against antigens of crude whole bacterial cell protein sonicate, lipopolysaccharide, flagella, and outer-membrane proteins. There was some evidence of an anamnestic response with IgA in bile following reinfection with the Salmonella. The peak response of antibody-producing cells from the spleens of infected chickens, assayed by solid-phase ELISA, occurred at 3 weeks postinoculation. A strong delayed hypersensitivity reaction, detected by foot-pad swelling after inoculation with either whole-cell or outer-membrane proteins, was observed between 2 and 5 weeks after infection with the Spcr mutant. The data indicate that outer-membrane proteins are major immunogens for both humoral and cell-mediated arms of the immune system.