Serological response of chickens naturally infected with Salmonella typhimurium detected by ELISA.

Four laying flocks of chickens in Britain, each with a history of Salmonella typhimurium infection, were investigated serologically and bacteriologically. Blood samples were taken from identified birds from a single house on each site and sent to the Central Veterinary Laboratory, Weybridge for serological examination using enzyme-linked immunosorbent assays (ELISA) and rapid slide agglutination test (RST) using stained S. pullorum. The identified birds were taken to the local Veterinary Investigation Centre for bacteriological examination. On site A no salmonellae were recovered from birds in the house chosen for serological examination. Of these birds approximately 20% had antibodies to S. typhimurium in ELISA which used either a lipopolysaccharide (LPS) or heat-extract (HE) antigen from S. typhimurium. S. typhimurium was recovered from birds in one other of the four houses on the same site; these birds were not tested serologically. On site B, S. typhimurium was isolated from 8% of the birds examined. Of the total tested serologically, a third to half were seropositive by S. typhimurium ELISA using the LPS and HE antigen respectively. A small proportion of birds was seropositive by S. enteritidis ELISA and RST. No salmonellae were isolated from the other two sites although about 10% of birds tested on site C were seropositive in S. typhimurium ELISA. Cross-reactions were seen between S. typhimurium antigens in the ELISA and experimentally prepared antiserum to S. enteritidis. The S. enteritidis ELISA was generally more specific although cross-reacting antibodies were detected in sera from birds on sites A and B.     

Effect of reticuloendotheliosis virus on the response of chickens to Salmonella typhimurium infection

Groups of 25 chickens free of maternal antibody to reticuloendotheliosis virus (REV) were inoculated with either third or seventh passage REV at either one or seven days of age. Some of the birds inoculated at day 1 with REV were inoculated with Salmonella typhimurium either concurrently or six or 13 days later while some of those inoculated with REV at day 7 were inoculated concurrently with S typhimurium. At day old, infection with S typhimurium alone caused the death of 12 of 25 chicks whereas in the dual infection, using the third passage REV, 18 of 25 birds died. Similarly no seven or 14 day old chickens died when challenged with S typhimurium alone, but previous day-old infection with REV caused a respective mortality of eight of 25 and five of 25 birds. With the seventh passage REV a similar pattern was seen. At day old S typhimurium infection alone killed seven of 25 birds whereas combined with virus the mortality was 14 of 25 and while S typhimurium alone killed none of 25 chicks infected at seven days old, the mortality in birds also infected with REV was 14 of 25. Combined virus and bacterial infections did not increase the proportion of feathering defects in birds surviving S typhimurium infections. There was a significantly higher proportion of feathering defects in birds infected with third passage virus compared with seventh passage virus. Although a higher proportion of birds had antibody responses to REV in the seventh than in the third passage group, there was no discernible difference in the effect the different viruses had on chickens' susceptibility to S typhimurium.     

Percloacal infection of chickens with Salmonella typhimurium

The characteristics of Salmonella typhimurium infection in hatchlings and adult poultry given by the intravenous, oral and percloacal routes were compared. Based on LD₅₀ determinations, 1-day-old birds were most susceptible to percloacal infection. It is claimed that this infection technique yields predictable results in terms of mortality and numbers of organisms in the liver, spleen and intestinal tissues. Though older birds do not die when given large doses of organisms by this route, small numbers are sufficient to establish transient infection or carriage in the intestine.Based on quantitative data, the course of the percloacal infection appeared similar to that given by the oral route in hatchlings; however, the number of organisms required to establish infection by the former route was significantly less. It is suggested that the percloacal infection route may be a useful model for studying Salmonella infection-immunity in birds, particularly if resistance mechanisms operating in the intestine are to be taken into account. The possibility that it may be an important infection route in natural circumstances must also be considered.     

Epizootiology of Salmonella typhimurium infection in chickens]

The incidence of S. typhimurium infections among fowl increased in thr region of Potsdam in general, and on various big farms in particular, 1976 and over the first half of 1977. The outbreaks included subclinical infections and clinically manifest diseases which caused remarkable loss of broilers from the affected stocks (up to 15.92 per cent). Parent stocks contaminated with S. typhimurium were to be the sources of infection in all cases. A total of 1,220 Salmonella strains were isolated from fowl and its environment, with 1,151 of them being S. typhimurium (2.98 per cent of all samples tested). The following amounts of S. typhimurium strains were isolated from different types of samples which had been collected from infected broiler stocks: 8.10 per cent from dead broilers, 5.86 per cent from dead broiler parents, 2.11 per cent from pulp linings of transport cages for day-old chicks, 1.23 per cent from litter, 1.0 per cent from hatching material (eggs or dead and jammed embryos, and 0.12 per cent from swabs used in hygiene supervision). No Salmonellae were isolated from feedstuff. The transmission of S. typhimurium, therefore, is though to have taken the route via the hatching egg and via congenitally infected chicks traded between breeders and propagation farms. The control and prophylaxis of S. typhimurium infections, therefore, should be based primarily on action in the centralised breeding stocks. Specific steps of such action are proposed. Fifty-three strains were biochemically and lysotypically analysed, with the following types being determined: ut/Ph 30 BT b, ut/Ph 30 BT c, n.c. 1/72/n.c. BT b, 2 n.c. BT a, and 1A/6 BT a. The first two types covered 84.9 per cent of all strains isolated from the fowl. All lysotype ut/Ph 30 strains isolated from fowl fell under the copenhagen variant which had rarely been isolated from man in the past. These results are likely to support the demand for a joint control programme for enteritis Salmonellae, with particular emphasis on S. typhimurium, for implementation in human and veterinary medicine.     

Detection of Salmonella typhimurium infection of chickens.

The serological response of two groups of chickens was followed by three techniques after experimental infection of one group with Salmonella typhimurium. Results obtained illustrate the greater sensitivity of the microantiglobulin (Coombs) test over more conventional methods for detecting salmonella agglutinins. The possibilities of the diagnostic use of the microantiglobulin test in the field are discussed.     

Factors influencing enhanced Salmonella typhimurium infection in Eimeria tenella-infected chickens

To investigate a possible mechanism involved in the enhancement of Salmonella typhimurium infection in chickens concurrently infected with Eimeria tenella, S typhimurium was given orally to chickens 7 days after E tenella inoculation. The number of viable S typhimurium decreased in the ceca of chickens not inoculated with E tenella, whereas the number gradually increased in the ceca of chickens inoculated with E tenella. Cecal contents were analyzed for pH value, oxidation-reduction potential, and amounts of short-chain fatty acids and bile acids. In the ceca of E tenella-inoculated chickens, the oxidation-reduction potential significantly (P less than 0.05) shifted to the oxidative phase, and the concentration of volatile fatty acids (acetic acid, propionic acid, and butyric acid) significantly (P less than 0.05) decreased. In both aerobic and anaerobic incubations, the number of viable S typhimurium in vitro decreased as the molar concentration of fatty acids increased. Experimental evidence indicated that multiplication of S typhimurium in the ceca of E tenella-inoculated chickens was associated with decreased concentrations of volatile fatty acids.     

The humeral immune response of swine after experimental infection with Salmonella typhimurium

LPS antibody reaction of S. Typhimurium was detected in a total of 111 two- to ten-week-old pigs, which had been exposed to experimental oral infection with this serovar in 16 separate experiments. The total antibody level was determined according to the meat juice ELISA provided for the Salmonella control programme and the antibody concentrations of the isotopes IgG1, IgG2, IgA and IgM. Although there were significant differences between the individual experiments with respect to the reaction intensity and proportion of the individual immune globulin isotypes, the following general statements on the dynamics of the antibody reaction can be made: Clearly elevated antibody levels could be observed one week after the onset of infection. After one and two weeks, respectively, 22.5 and 52.9% of the infected animals showed antibody levels which were to be evaluated as positive in the sense of the Salmonella control programme (> or = 40%). While the concentration of the IgM-related antibodies decreased again during the second week already, the antibody concentration of IgG1 and IgG2 continued to increase until the end of the experiment after 3 weeks. The clear differences in the reactions between the experiments cannot be explained solely by defined conditions of infection such as strain of infection, infective dose or age of the pigs. Obviously, other non-defined or non-recognized factors may have a decisive influence on the dynamics of the antibody reaction in the animal groups.     

Infection and immune responses in chickens exposed to Salmonella typhimurium

The transmission of Salmonella typhimurium among adult chickens maintained in wire-floored cages was examined. Within 6 days, organisms had spread from infected seed chickens to the majority of uninoculated chickens. Initially, organisms were found in the intestinal tract alone, but by day 20, all chickens were infected, and most of these had organisms in the liver, spleen, and intestinal tract. Antibodies to whole bacterial cells and lipopolysaccharide were detected in the serum and bile of all chickens from day 24. Clearance of organisms from the tissues was not apparent until after day 33. Cell-mediated immunity to salmonella antigens was also induced in chickens that naturally acquired the infection. The role of these parameters of immunity in the clearance of organisms is discussed.     

Biological control of Salmonella typhimurium in young chickens

The effect of dietary lactose and anaerobic cultures of cecal microflora of mature chickens on the colonization of young broiler chickens by Salmonella typhimurium was evaluated. Newly hatched chicks were given either no treatment (controls), anaerobic cecal cultures, lactose (2.5%) in the drinking water, or both anaerobic cultures and lactose. Chicks were challenged per os at 3 days of age with either 10(6) or 10(8) S. typhimurium resistant to nalidixic acid and novobiocin. On day 10, the cecal contents of the chicks were examined for S. typhimurium, pH, short-chained volatile fatty acids (VFAs), undissociated VFAs, and lactic acid. Chicks given either lactose alone or cecal anaerobes alone had significantly (P less than 0.05) fewer S. typhimurium recovered from their ceca than the controls. Chicks given the combination of dietary lactose and cecal anaerobes had significantly fewer S. typhimurium recovered from their ceca than the chicks given dietary lactose or cecal anaerobes alone. Chicks given lactose had significant (P less than 0.05) increases in the lactic acid concentration of their cecal contents. Increased lactic acid concentrations were directly correlated to decreased cecal pH values and caused a reduction in the total concentration of VFAs but a significant (P less than 0.05) increase in the undissociated form of some VFAs.     

Assessment of attenuated Salmonella vaccine strains in controlling experimental Salmonella Typhimurium infection in chickens

Salmonella hold considerable promise as vaccine delivery vectors for heterologous antigens in chickens. Such vaccines have the potential additional benefit of also controlling Salmonella infection in immunized birds. As a way of selecting attenuated strains with optimal immunogenic potential as antigen delivery vectors, this study screened 20 novel Salmonella Typhimurium vaccine strains, differing in mutations associated with delayed antigen synthesis and delayed attenuation, for their efficacy in controlling colonization by virulent Salmonella Typhimurium, as well as for their persistence in the intestine and the spleen. Marked differences were observed between strains in these characteristics, which provide the basis for selection for further study as vaccine vectors.     

Passive protection of calves against experimental infection with Salmonella typhimurium

Cows were vaccinated with formalin-killed Salmonella typhimurium approximately seven weeks and two weeks before parturition to investigate whether passive immunity could protect their calves against experimental S typhimurium infection. After birth the calves were left with their dam for 48 hours and then separated and fed cold, stored colostrum from their own dam for a further eight days. Oral challenge five days after birth with 10(8) S typhimurium did not result in the death of these calves even when they had absorbed little colostrum. Mortality was reduced to 22 per cent in calves which sucked from vaccinated dams and were then fed colostrum from unvaccinated cows and to 50 per cent in calves born to unvaccinated cows and later fed colostrum from vaccinated animals. Calves which sucked from a vaccinated dam and then received stored colostrum from the same cow excreted salmonellas for significantly shorter periods after challenge and were less often infected at necropsy 28 days after inoculation. Protection was not correlated with the levels of O or H agglutinating antibodies in serum, which were at a maximum 24 hours after sucking and then slowly declined. There was no evidence of an active antibody response in the serum. Measurement of the O and H response of cows after vaccination indicated that the vaccination schedule could be improved. The highest levels of agglutinating antibody were measured between two and three weeks after the first vaccination and there was only a minimal response to the second vaccination before parturition.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effect of T-2 toxin on resistance to systemic Salmonella typhimurium infection of newly hatched chickens

Newly hatched chickens were treated with the trichothecene mycotoxin, T-2 toxin, during the first day of life. Control chickens were treated with other agents known to cause immunosuppression--cyclosporine, cyclophosphamide, and aflatoxin. Chickens were infected on day 6 (5 days after treatment with T-2 toxin) by intraperitoneal inoculation with Salmonella typhimurium. Blood samples were collected from treated chickens (noninfected) and used to assess the responsiveness of blood lymphocytes to T-cell or B-cell mitogens, phytohemagglutinin, or lipopolysaccharide, respectively. The T-2 toxin had a profound negative effect on the ability of the chickens to resist salmonellosis, as measured by survival. However, the toxin effect in reducing phytohemagglutinin- and lipopolysaccharide-stimulated mitogenesis, though significant (P greater than 0.05), was not severe. Our data indicate a direct effect of T-2 toxin on native resistance to systemic salmonellosis, which was not accompanied by marked alteration in T- or B-cell responses to mitogenic stimulation.     

Testing a bivalent vaccine in sheep against experimental infection with Salmonella bovismorbificans and Salmonella typhimurium

Abstract

Extract

Increasing awareness of the threat from salmonellosis to animal and human health, and recognition of implications to the farming economy and meat marketing industry in New Zealand, have emphasized the need for effective measures for controlling this disease. As pointed out by Beckett (1967 Beckett, F. W. 1967. The use of Salmonella vaccine in outbreaks of salmonellosis in sheep. N.Z. vet. J., 15: 66–69. [Taylor &; Francis Online] , [Google Scholar]), reduction in stocking rates or use of chemotherapeutic agents is often impracticable or ineffective. Other recent publications, drawing attention to the possible role of vaccination in controlling salmonellosis, have dealt with fundamental aspects of immunity (Jonas 1967 a Jonas, W. E. 1967a. Studies of the immunological aspects of salmonellosis of mice: active immunity. N.Z. vet. J., 15: 27–30. [Taylor &; Francis Online] , [Google Scholar]; b Jonas, W. E. 1967b. Salmonellosis of sheep: use of serum from a vaccinated ewe in a mouse passive protection test. N.Z. vet. J., 15: 55–61. [Taylor &; Francis Online] , [Google Scholar]) and with evaluation of an experimental vaccine under field conditions (Wallace and Murch, 1967 Wallace, G. V. and Murch, O. 1967. Field trials to assess the value of a bivalent killed salmonella vaccine in the control of ovine salmonellosis. N.Z. vet. J., 15: 62–65. [Taylor &; Francis Online] , [Google Scholar]).