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Biomedical subjects

P A Barrow

Publications and source records attributed to P A Barrow.

At least 55 records · Page 3Linked to original sources
PubMed1996

World Health Organisation--supervised interlaboratory comparison of ELISAs for the serological detection of Salmonella enterica serotype Enteritidis in chickens.

A collaborative exercise, supervised by the World Health Organisation, was set up to compare ELISAs used for the serological detection of Salmonella enteritica serotype Enteritidis in chickens. The aim was to ascertain how far agreement could be reached on the interpretation of optical density readings for high titre, intermediate titre and low titre sera. Two sets of sera were sent to 14 participants. The first set compared high, medium and low titre sera raised in specified-pathogen-free and commercial broiler breeder chickens. The second set comprised 20 sera of different antibody titres raised in commercial birds reared under laboratory conditions and sent blind. Both indirect and double-antibody sandwich blocking ELISAs were used with a number of different detecting antigens. With a few exceptions good agreement was reached on the interpretation of results obtained from high and low titre sera from the optical density obtained with a single serum dilution. Differences were observed in the interpretation of medium titre sera. The results suggested that most ELISAs produce reasonably comparable results and that practical problems may arise from interpretation of the results mainly as a result of the choice of the criteria used for differentiating sera obtained from infected and uninfected chickens. These problems are discussed.

Animals↗
PubMed1996

Growth suppression in early-stationary-phase nutrient broth cultures of Salmonella typhimurium and Escherichia coli is genus specific and not regulated by sigma S.

We have studied the growth suppression seen in early-stationary-phase LB broth cultures of Salmonella typhimurium. Multiplication of small numbers of an antibiotic-resistant S. typhimurium mutant was prevented when the mutant was added to 24-h cultures of the antibiotic-sensitive parent strain, whereas an antibiotic-resistant mutant of an Escherichia coli strain added to the same culture grew well. A 24-h E. coli culture produced a similar specific bacteriostatic inhibition against E. coli. In older cultures, a specific bactericidal effect similar to that observed by M. M. Zambrano and R. Kolter (J. Bacteriol. 175:5642-5647, 1993) was also observed. Whether incubated statically or shaken, sufficient nutrients were present in the filtered supernatants of 24-h cultures for small inocula of the same strain to multiply to ca. 10(9) CFU/ml after reincubation. Introduction of the rpoS mutation had no effect on the specific bacteriostatic inhibition. Similar specific inhibition was also observed in strains of Citrobacter freundii, Klebsiella pneumoniae, Enterobacter agglomerans, and Shigella spp. Experiments in which the 24-h culture was physically separated from the antibiotic-resistant mutant by using a dialysis membrane were carried out. These results indicated that the inhibition might be mediated by a diffusible but labile chemical mediator.

Bacterial Proteins↗
PubMed1996

Reduction in incidence of experimental fowl typhoid by incorporation of a commercial formic acid preparation (Bio-Add) into poultry feed.

The experiment described evaluated the effect of a commercial in-feed preparation (Bio-Add) involving a mixture of formic acid and propionic acid on the incidence of experimental fowl typhoid in groups of 41 and 42 1-wk-old Rhode Island Red chickens. The chickens were infected through contact with 12 identical chickens that had been inoculated orally with 10(8) cfu of Salmonella gallinarum strain 9. The incidence of mortality and morbidity due to fowl typhoid was 31/41 (76%) in birds given untreated feed and 14/42 (33%) in birds given feed treated with Bio-Add.

Animals↗
PubMed1996

[Inhibition phenomena between Salmonella strains--a new aspect of salmonella infection control in poultry].

Freshly hatched chickens show a very high susceptibility to Salmonella infections and control measures are therefore frequently focused on the period shortly after hatching. Experimental investigations using one strain against itself, differentiated by different antibiotic resistance markers, have shown that colonisation with Salmonella prevents the establishment of subsequently inoculated challenge organisms in the chicken gut. The inhibition effect lasts for several days and is detectable even when a challenge dose of 10(8) organisms is used. It is dependent of the breed of bird. Chickens colonised with Salmonella shed a subsequently inoculated challenge strain with significant lower numbers for several weeks than do non colonised control birds. The phenomenon is strain specific but not serovarspecific as has been shown in investigations using different strains of the same and other serovars for colonisation and challenge. The phenomenon shows a large variability between strains. Using other Enterobacteriaceae strains comparable inhibition against Salmonella was not observed. One important topic for further investigation is the capability of Salmonella live vaccines given orally to establish a protection effect, based on the inhibition phenomenon in the first few days of live, developing into a long-lasting immunity when birds reach immunological maturity.

Animals↗
PubMed1995

Interference between Salmonella serotypes in intestinal colonisation of chickens: correlation with in vitro behaviour.

The effect of colonisation of the alimentary tract of newly hatched chicks by different Salmonella serotypes on the establishment in the gut by other Salmonella strains inoculated afterwards was assessed. Although profound inhibition of colonisation had been found previously to be genus-specific, considerable variation was found within the Salmonella genus. Some strains were found to be much more inhibitory than others and some were more easily inhibited than were others. There was not an absolute relationship between inhibitory activity and colonisation ability. No relationship was seen between inhibition and serotype or phage types within serotypes. There was no correlation between in vivo inhibition and the extent of inhibition that occurred in early stationary phase cultures in rich, undefined broth cultures.

Animals↗
PubMed1994

Serological diagnosis of Salmonella serotype enteritidis infections in poultry by ELISA and other tests.

Serological methods have increasingly been used for the detection of invasive Salmonella serotypes including enteritidis in poultry. Different types of ELISA, particularly indirect or double antibody-blocking assays using a variety of antigens such as lipopolysaccharide, flagella and SEF14 fimbrial antigen are used as part of control programmes in a number of countries. There are many advantages to using such assays for preliminary screening of flocks prior to using bacteriological culture methods.

Agglutination Tests↗
PubMed1994

Comparison of multiplex PCR and standard bacteriological methods of detecting Salmonella on chicken skin.

A multiplex PCR technique was compared with standard bacteriological techniques for the detection of Salmonella, and Salm. enteritidis in particular, in samples of chicken skin obtained from retail outlets. Five of 68 samples were positive by both systems (7.4%), 58 of 68 were negative by both systems (85%), five were positive by PCR only (7.4%) but none were positive by culture only. The PCR was quicker and more sensitive than the bacteriological methods, finding 15% of samples positive for Salmonella compared to 7.4%, and taking less than 24 h to reach a result as opposed to 2-3 d.

Animals↗
PubMed1994

Host specificity of Salmonella infection in chickens and mice is expressed in vivo primarily at the level of the reticuloendothelial system.

By experimental infection, host-specific Salmonella serotypes were shown to demonstrate specificities for chickens, mice, and other laboratory animals. Following oral inoculation, four strains of Salmonella gallinarum and two S. pullorum strains, isolated from diseased poultry, were more virulent for chickens than for mice. By contrast, four strains each of S. choleraesuis and S. dublin, isolated from diseased pigs and cattle, respectively, were more virulent for mice than for chickens. These results were also reflected in the degree of virulence expressed after parenteral inoculation. In addition, S. choleraesuis, but not other serotypes, killed rats, guinea pigs, and rabbits. S. typhimurium strains varied widely in their virulence, and some strains were virulent for both mice and chickens. Four other serotypes isolated from poultry or human food poisoning cases and a nonpathogenic Escherichia coli strain were much less virulent for both experimental host species. Most of the host-specific Salmonella serotypes studied were able to colonize the distal alimentary tract and invade the tissues in both mice and chickens to various degrees. There was, however, a greater difference in the ability to survive and multiply in the visceral organs, particularly the spleen and the liver, once invasion had occurred which correlated with the virulence for the host species involved.

Animals↗
PubMed1993

Construction of an aroA mutant of Salmonella serotype Gallinarum: its effectiveness in immunization against experimental fowl typhoid.

An aroA mutant was produced from a virulent strain of Salmonella Gallinarum, the causative agent of fowl typhoid in poultry, by Tn10 insertional inactivation and deletion. The mutant was highly attenuated for chickens. A single intramuscular immunization of 2-week-old chickens with 10(7) mutant organisms reduced mortality following oral challenge with 10(8) organisms of the parent strain from 63 to 30%. In a second experiment multiple immunizations with 10(8) mutant organisms reduced mortality after challenge with 10(7) organisms of the virulent strain from 77 to 0%. By oral inoculation the mutant was not able to immunize chickens against oral challenge.

Animals↗
PubMed1993

The role of a 54-kb plasmid in the virulence of strains of Salmonella enteritidis of phage type 4 for chickens and mice.

The role of a 54-kb plasmid in the virulence of Salmonella Enteritidis strains of phage type 4 was examined in mice, young chickens and adult laying-hens. Although the plasmid was essential for full expression of virulence in mice, differences in expression of virulence by this 54-kb plasmid were noted among strains; thus, introduction of the plasmid to a naturally occurring strain that lacked it did not make that strain virulent. In newly hatched chickens, virulence of an Enteritidis strain of phage type 4 by oral or parenteral routes was unrelated to possession of this plasmid which, similarly, played no role in infection in egg-laying hens. When a strain of Enteritidis phage type 4 and a plasmid-cured strain derived from it were given orally to chickens, both strains were equally invasive and their patterns of localisation in spleen, liver and ovaries were similar and they were isolated at similar frequencies from cultured, laid eggs.

Animals↗
PubMed1992

Further observations on the serological response to experimental Salmonella typhimurium in chickens measured by ELISA.

An indirect ELISA developed for the serological detection of Salmonella typhimurium in chickens using lipopolysaccharide as detecting antigen has been evaluated further in experimental infections. Following oral infection of 24-week-old laying hens with an invasive strain of S. typhimurium, high titres of specific circulating IgG were induced which were maintained for 20 weeks. Similar IgG titres were found in egg yolk. When 4-day-old chickens were infected high antibody titres persisted for 45 weeks. Chickens inoculated orally or intramuscularly with different numbers of S. typhimurium organisms showed graded serum IgG responses to LPS. The IgG titres in experimentally infected in-bred lines of chickens which showed greater genetic resistance to salmonella infection were significantly lower than those found in more susceptible lines. Oral and intramuscular infection with 18 different types of enterobacteria, including avian pathogenic E. coli, Citrobacter spp., Klebsiella spp., Proteus spp. and citrobacter-like organisms possessing some salmonella LPS (none possessed the O-4 antigen) and flagella antigens, did not induce S. typhimurium LPS-specific IgG responses. Chickens infected orally with rough or non-flagellate mutants of S. typhimurium did not induce high titres of LPS or flagella-specific IgG respectively. Sera obtained from S. typhimurium-infected chickens showed much higher titres against S. typhimurium LPS than with those antigens from other serotypes, including S. enteritidis.

Animals↗
PubMed1992

In-vitro and in-vivo characteristics of TnphoA mutant strains of Salmonella serotype Gallinarum not invasive for tissue culture cells.

Insertion mutants of a strain of Salmonella serotype Gallinarum, the cause of fowl typhoid, were produced with transposon TnphoA. Eight mutants were identified as being less invasive in cell culture than the parent strain. Neither the parent strain nor the mutants showed mannose-sensitive haemagglutination of various red blood cell species. Although two mutants gave mannose-resistant (MR) haemagglutination of red cells of different animal species these MR activities could not be correlated with other characteristics in vitro or in vivo. The mutant strains were divisible into three classes by their patterns of invasiveness and adhesiveness in vitro and by changes in membrane proteins. Strains of classes 1 and 2 had single transposon insertions, detected by Southern hybridisation, of 8.9 and 2.4 kb EcoRV-digested chromosomal-fragments, respectively, and were slightly less invasive than the parent strain. They were no less virulent for chickens by the oral route than a mutant strain shown to be as invasive in vitro as the parent strain. The class-2 mutant was also less adhesive than the other strains. The single mutant strain of class 3 which contained insertions in several chromosomal fragments, was non-invasive in Vero cells (less than log10 1.0 cfu/ml recovered compared with log10 2.88 for the parent strain) and showed reduced virulence by both oral and intramuscular routes. The mutant strains of all classes were taken up equally rapidly from the blood by the spleen. Intramuscular immunisation of chickens with the class-3 mutant strain gave complete protection against oral challenge 3 weeks later with 1000 oral LD50 doses of the virulent parent strain.

Animals↗
PubMed1991

The activity in the chicken alimentary tract of bacteriophages lytic for Salmonella typhimurium.

Bacteriophages lytic for Salmonella typhimurium were isolated in considerable numbers from chickens experimentally infected with S. typhimurium, and in much lower numbers from the chicken feed. Lytic phages were also regularly isolated from human sewerage systems. One of these was used to inoculate S. typhimurium--infected two day-old chickens orally and via the feed. The phage took longer to establish in the caeca than did the Salmonella and it disappeared when the caecal S. typhimurium counts fell to 10(6) CFU/ml. No neutralizing antibodies to the phage were detected in the serum of these chickens. In a second experiment, five of 30 chickens similarly infected with S. typhimurium were inoculated with the phage. Within 3 days, the phage was isolated from 72% of the "in-contact" birds. A second phage, isolated from sewage, when inoculated into newly-hatched chickens simultaneously with any of 3 strains of S. typhimurium, produced a considerable reduction in mortality in the birds. This effect was only produced by inoculation of high concentrations of phage (greater than 10(10) PFU/ml). The phage produced reductions in the viable numbers of S. typhimurium in the crop, small intestine and caeca for up to 12 h after inoculation, with smaller reductions in bacterial numbers in the liver at 24 and 48 h after infection.

Animals↗
PubMed1991

In vitro characterization of intra-generic inhibition of growth in Salmonella typhimurium.

The intra-generic inhibition of bacterial growth observed previously in vivo and in vitro with strains of Salmonella, Citrobacter and E. coli was studied in vitro using S. typhimurium strain F98. There was complete inhibition of multiplication of S. typhimurium when it was added to stationary-phase broth cultures of different Salmonella serotypes, but only partial inhibition when added to broth cultures of E. coli. The degree of inhibition between different mutants of F98 was affected by the numbers of bacteria of the inhibiting strain, but this was not the only factor, since exponential-phase bacterial cells were less inhibitory than stationary-phase cells. The inhibitory effect was produced at temperatures between 20 degrees C and 40 degrees C. The complete inhibition of growth observed between F98 mutants was abolished by ampicillin, rifampicin and streptomycin, but not by nalidixic acid. Inhibition was also prevented by separating the two cultures by a dialysis membrane. A TnphoA insertion mutant of F98 was produced which did not show inhibition in vitro but was still inhibitory in vivo. It is suggested that this complete inhibition of bacterial multiplication between organisms of the same genus, which is greater than that produced between organisms from different genera, is mediated by a cell surface protein.

Animals↗
PubMed1990

Antibody response to experimental Salmonella typhimurium infection in chickens measured by ELISA.

An indirect ELISA has been developed to detect Salmonella typhimurium antibodies in chicken sera, using whole bacterial cell protein, flagellar protein or lipopolysaccharide as antigens. In experimental infections high concentrations of S typhimurium-specific IgG persisted after the faecal excretion of S typhimurium had ceased, whereas the specific IgM response was transitory. Some uninfected chickens placed in contact with experimentally infected birds developed high IgG titres in the absence of detectable faecal excretion. Other S typhimurium strains, which varied in their invasive abilities, also induced high titres of IgG. The ELISA allowed chickens infected experimentally with S typhimurium to be differentiated from chickens infected with 10 other serotypes, including S enteritidis. The use of whole blood in place of serum in the ELISA reduced the titres slightly. The storage of serum dried on to filter paper strips for four weeks produced little change in ELISA antibody titre, and the treatment of such strips with phenol or chloroform vapour had little or no effect on the antibody titre.

Animals↗