A disc ELISA for the detection of salmonella group D antibodies in poultry

An ELISA using lipopolysaccharide antigens prepared from Salmonella gallinarum and S enteritidis was developed for the serological diagnosis of fowl typhoid and S enteritidis infection in poultry. There was good agreement between the results of the ELISA and conventional serological tests when samples from naturally infected birds and S enteritidis immunised birds were tested. Some cross reactions were observed when serum samples from S typhimurium infected birds were tested by ELISA. Subsequently a disc ELISA, using filter paper discs, was developed to facilitate sampling and testing of poultry. There was good correlation between the results of the disc and serum ELISAs and the test is recommended for the field testing of birds.     

Growth of Salmonella typhimurium in the caecum of gnotobiotic chickens with Eimeria tenella

Gnotobiotic chickens infected with Eimeria tenella (5 X 10(4) oocysts per bird) received an oral inoculation of 100 Salmonella typhimurium two, four, six or eight days after coccidial infection at four days old. When S typhimurium was given two or four days after E tenella infection, S typhimurium counts in the caecal contents were similar to the counts in birds infected with S typhimurium alone. When S typhimurium was given six or eight days after E tenella infection, counts of the organism were significantly greater than with S typhimurium infection alone. There were no differences in the number of chickens positive for S typhimurium in the caecal contents, bile, liver and spleen between the two groups.     

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.     

Epidemiology of Salmonella typhimurium infection in calves: excretion of S typhimurium in the faeces of calves in different management systems

Twenty-five batches of market-purchased calves, comprising 589 animals on 11 farms, were examined for the excretion of salmonellas for at least 28 days after their arrival. Salmonellas were found in 217 (51 per cent) of 423 calves from 18 of 21 batches on nine farms. The calves were housed in single pens on eight farms and in groups on the others. The trends in new excreters and the proportion of Salmonella typhimurium excreters were similar in both types of housing, a peak of excretion which was slightly higher in single penned calves, being reached 18 to 19 days after arrival. New excreters were found at most samplings in both types of housing, but the peak of new excreters appeared earlier and declined sooner in single penned calves. On average, single penned animals excreted salmonellas longer. Salmonella dublin was found only in single-penned calves, and the trend in excretion was similar to that for S typhimurium in single-penned calves except that the peak was at about 28 days. Prophylactic feeding of antibiotics was practised on nine farms, but appeared to have no influence on the excretion of salmonellas. Salmonellas were isolated from the environment in six of nine farms studied, even after cleaning and disinfection. It is suggested that the persistence of salmonellas in the environment deserves more attention in the formulation of control programmes.     

The occurrence of salmonellae in wild birds (Passeriformes, Lariformes)

The occurrence of salmonellae was investigated in 1011 cloacal smears of 21 species of wild birds caught at the locality of Hermanicky rybník near Ostrava. Salmonella typhimurium serotype was isolated from one of the 23 laughing gulls (Larus ridibundus) which were examined; this corresponds to a 4.3% invasion frequency. The result of the examination of all other birds was negative. On the basis of the results of this study and the results published by other authors, laughing gull can be considered as a possible source of salmonellae for farm animal stocks, particularly water fowl. It is recommended that laughing gull should be taken into consideration when seeking, for diagnostic purposes, the sources of salmonellae from which water fowl are invaded.     

Pasteurella multocida in wild mammals and birds in California: prevalence and virulence for turkeys

Samples collected from the oropharynx of wild mammals and birds trapped on 36 turkey farms in California were evaluated for the presence of Pasteurella multocida. A total of 966 animals were collected from 18 premises that had experienced an outbreak of fowl cholera within the past 2-8 months; samples were collected from 16 of these 18 premises within 2-8 weeks of outbreak notification and while the infected flock was still present. A total of 939 animals were trapped from an additional 18 premises that had not reported any outbreaks of fowl cholera within at least 4 months, if ever. Forty-eight isolates of P. multocida, of a variety of somatic serotypes, were recovered from 6 species of mammals and 3 species of birds. On only 2 of 7 premises was the somatic serotype of the isolates obtained from wildlife the same as the isolate obtained from tissues of turkeys that had died of fowl cholera on the same premises. Tests for virulence to turkeys were conducted with 31 of the isolates. Seventeen of these isolates caused mortality in turkeys. Wide ranges in mortality rates and median times to death were observed.     

Reciprocal competitive exclusion of salmonella and Escherichia coli by native intestinal microflora of the chicken and turkey

Both the native intestinal microflora of chickens that protected chicks against salmonellae and Escherichia coli and native turkey intestinal microflora were evaluated for their reciprocal protective capacity in both species against Salmonella typhimurium and a pathogenic strain of E. coli. Nalidixic-acid-resistant forms of the S. typhimurium and E. coli strains were used in seeder-bird and individual-bird challenge tests. Reciprocal protection was provided by native chicken and turkey intestinal microflora in chicks and poults against S. typhimurium and the pathogenic strain of E. coli. The chicken and turkey microflora appeared to be equally effective in protecting the two species from S. typhimurium, but protection against E. coli was somewhat greater in the chicken than in the turkey.     

The isolation of salmonellae, Newcastle disease virus and other infectious agents from quarantined imported birds in Canada.

Necropsy and culture results are presented for 269 consignments of imported birds (mainly psittacine and passerine species) examined between January 1977 and August 1980. Consignments were submitted for diagnosis of clinical illness or deaths occurring among these birds while they were in quarantine before entry into Canada. Enteritis and injury were the most frequent diagnoses. Pathogens or potential pathogens were isolated from 77% of consignments. Newcastle disease virus was isolated nine times, and Chlamydia psittaci was isolated once. Escherichia coli (from 113 consignments) and salmonellae (from 49) were the most common bacteria isolated, and reoviruses (from 22) and paramyxoviruses other than Newcastle disease virus (from 22) were the most common viruses. Salmonella typhimurium was the most common Salmonella serovar. Salmonella hadar was isolated from turkey poults imported from Great Britain. The possible public health significance of the role of imported birds in the introduction of exotic Salmonella serovars, or of serovars resistant to several antimicrobials is discussed.     

Effects of cage contamination with coccidia and salmonella on acute salmonellosis in young chickens

The effects of concurrent cage contamination with Salmonella typhimurium and Eimeria tenella on the establishment of salmonella infection in day-old chickens were investigated. Chickens were divided into five groups: uninfected recipient birds placed in a cage contaminated by donor birds infected with E. tenella and S. typhimurium; E. tenella-infected recipients placed in a cage contaminated by S. typhimurium-infected donors; uninfected recipients placed in a cage contaminated by S. typhimurium-infected donors; E. tenella-infected recipients placed in a cage contaminated by uninfected donors; and uninfected recipients placed in a cage contaminated by uninfected donors. Three identical trials were conducted. Recipient birds were necropsied 4, 7, and 11 days after caging. In the cage where donor birds infected with both organisms had been reared, S. typhimurium counts in feces and number of feces positive for S. typhimurium were significantly (P less than 0.05) greater than those in the other cages on days 0, 4, and 7 after caging. Moreover, in this cage, more chicks died, counts of S. typhimurium in cecal contents were greater, and more birds were positive for S. typhimurium than in the other groups. This suggests that S. typhimurium infection in day-old chickens is enhanced in cages contaminated with E. tenella and S. typhimurium compared with infection in cages contaminated with S. typhimurium alone.     

Development and application of an ELISA for detecting antibodies to Salmonella enteritidis in chicken flocks.

Enzyme-linked immunosorbent assays (ELISAs) were developed for the detection of IgG antibody to Salmonella enteritidis in poultry flocks. A lipopolysaccharide (LPS) and heat-extracted (HE) antigen for use in the ELISA were evaluated together with the rapid slide test (RST), microagglutination test (MT) and the microantiglobulin (MAG) test. In experimentally infected specific pathogen free chickens, good correlation was seen between all tests although, generally, the MT and MAG detected antibody earlier and titres peaked earlier than the ELISAs. The LPS antigen detected antibody earlier than the HE antigen but the latter gave higher titres in the later stages of infection. Cross reactions were seen between S enteritidis and S typhimurium in the ELISAs although homologous reactions were always much higher. Antisera to S montevideo or S senftenberg gave weak positive reactions in both S enteritidis ELISAs. Serological and bacteriological examinations of representative samples from two commercial chicken flocks were carried out. In flock A the HE-ELISA and MAG test detected antibody in nearly all birds. The LPS-ELISA detected antibody in over 60 per cent of birds, while the MT and RST detected few seropositive birds. The whole blood test using the stained S pullorum antigen on the farm detected antibody in just under 25 per cent of the birds. S enteritidis was isolated from the organs of 25 per cent of the birds. All birds in flock B were seronegative by all tests; no salmonellae were isolated from the organs of these birds.     

The serologic response to Salmonella enteritidis and Salmonella typhimurium in experimentally infected chickens,followed by an indirect lipopolysaccharide enzyme-linked immunosorbent assay and bacteriologic examinations through a one-year period

Three groups of 100 individually marked salmonella-free chickens were followed for a period of 53 wk. The chickens were infected as day olds by crop instillation of 10(8) colony-forming units: one group with Salmonella enteritidis and a second group with Salmonella typhimurium. A third group was kept uninfected as controls. The groups were monitored bacteriologically by examination of cloacal swabs and organs and serologically by examination of serum and egg yolk by a lipopolysaccharide enzyme-linked immunosorbent assay throughout the period. Within the first week, 100% of birds in both infected groups were excreting salmonella bacteria in the feces. However, the number of fecal excretors declined rapidly with time, down to 6% in 16 wk for S. typhimurium and down to a similar level within the first 8 wk for S. enteritidis. For the latter, relapses with up to 40% positive birds were observed at the onset of egg production. For both S. typhimurium and S. enteritidis, positive bacteriologic cultures were obtained by sampling from internal organs at the end of the experiment, more than 1 yr from the time of infection. At the age of 6-7 wk, 50% of the chickens in the two infected groups showed a measurable serologic response in serum samples. The response persisted throughout the study in both serum and egg yolk samples. The inclusion of serologic methods is a valuable additional tool in the detection of salmonella in poultry, but serology should be used in conjunction with bacteriologic methods in surveillance programs, in particular to detect flocks in early stages of infection before a measurable serologic response has been raised.     

Serological and bacteriological investigations of chickens from flocks naturally infected with Salmonella enteritidis

Groups of 10 birds were obtained from four flocks which had shown evidence of natural salmonella infection. S enteritidis had been isolated from three flocks and S typhimurium from the fourth. Each bird was housed in a separate cage and blood samples and cloacal swabs were taken weekly to follow the course of natural infection. After four weeks the birds were killed and examined post mortem. The isolation of Salmonella species could not be related to the serological results. In individual birds the rapid slide test and tube agglutination test could not be relied upon to detect infection; the microantiglobulin test and the enzyme-linked immunosorbent assay (ELISA) were more sensitive than the other tests and detected some infected birds that were negative by the rapid slide and tube agglutination tests, and also showed high titres in some birds from which Salmonella species could not be isolated post mortem. Sera obtained from two flocks which had a history of natural S enteritidis infection were evaluated by all the tests; evidence of infection was found with the microantiglobulin and ELISA tests but not with the other tests.     

Therapeutic trials with native intestinal microflora for Salmonella typhimurium infections in chickens

Chicks infected with Salmonella typhimurium at 1 day of age were treated with normal intestinal flora starting 3 or 9 days later. Chicks were reared on the floor in isolated pens with wood shavings for litter. Cloacal-swab cultures were the main criteria for determining S. typhimurium infection. A single treatment reduced the infection period, but multiple treatments hastened the process considerably: after 6-8 weeks, birds given multiple treatments yielded sporadic or no isolations of S. typhimurium. It appears that litter from birds colonized by a protective microflora may serve as a vehicle for transmitting the flora to succeeding groups of birds.     

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 antibody to Salmonella enteritidis and S typhimurium in the yolk of hens' eggs

Enzyme-linked immunosorbent assays (ELISAs) have been developed to detect IgG antibodies to Salmonella enteritidis and S typhimurium in the yolk of hens' eggs. Better discrimination and more consistent results were obtained between eggs from experimentally infected and uninfected hens by using saline-dilution of yolk rather than chloroform extraction. Threshold absorbance values were determined in three salmonella-free flocks, and on the basis of these results ELISA optical density values greater than 0.25 were considered to be positive for antibodies to salmonella. Four flocks with a history of salmonella infection were examined; three contained birds which were seropositive for S enteritidis by ELISA and from which S enteritidis was isolated, and a large proportion of eggs from these birds contained antibody to S enteritidis. Eggs from the fourth flock had no detectable antibody, although serum antibody was detected in some birds. No salmonellae were isolated from the yolks of the eggs from any of the four flocks.     

斑点免疫金渗滤法检测鸡白痢/鸡伤寒沙门氏菌抗体的应用研究

将鸡伤寒沙门氏菌LPS抗原包被于渗滤盒检测区（T区），针对沙门氏菌O9抗原单克隆抗体3-47-0包被在质控区（C区），用胶体金标记鸡伤寒沙门氏菌LPS抗原，建立了一种抗原夹心法快速检测鸡白痢、鸡伤寒沙门氏菌抗体的斑点免疫金渗滤法（DIGFA）。DIGFA比玻板凝集试验（PAT）灵敏度高，人工感染鸡试验表明在感染后第7d即可从32只鸡中的7只检测到抗体，在时间上早于PAT。722份现场血清样品的检疫结果表明，DIGFA的阳性检出率稍高于PAT，阳性样品抗体几何平均滴度DIGFA大于PAT法（P〈0．05）。DIGFA的结果得到细菌分离试验的验证。这些结果表明DIGFA快速、灵敏、准确，为鸡伤寒和鸡白痢的检疫提供了一个新的有效手段。     

Prevalence and antimicrobial resistance of Salmonella spp. in pet mammals, reptiles, fish aquarium water, and birds in Trinidad

The prevalence of Salmonella spp. was determined in 970 animals comprising 423 pet birds, 485 fish aquaria water and 62 other pets (40 pet mammals, 14 reptiles, eight others - crustaceans, snail, stingray) from both pet shops and households throughout Trinidad. The serotypes of Salmonella spp. isolated were identified and the resistance to various antimicrobial agents was determined. Overall nine (0.9%) of 970 pet animals were positive for Salmonella spp. Six isolates of Salmonella spp. were recovered from all pet birds with two isolates of serotype Aberdeen and one isolate each of Thompson, Rubislaw, Panama and Newport. The prevalence of Salmonella spp. in birds was 0.9%. Four isolates of Salmonella spp. were recovered from fish aquaria water, serotypes included Panama (two isolates), Newport (one isolate) and Virchow (one isolate). Prevalence of Salmonella spp. from fish aquaria was 0.4%. No isolate of Salmonella spp. was detected in pet mammals sampled while two isolates were recovered from reptiles, S. Enteritidis and S. Montevideo. One isolate of Salmonella spp. was recovered from a stingray, serotype unknown. Antimicrobial resistance was present is all animal types. The highest prevalence of resistance was to streptomycin among isolates from birds (83.3%) and other pets (100.0%) while isolates from fish aquarium water exhibited comparatively high resistance to cephalothin (50.0%). It was concluded that the isolation of Salmonella spp. from apparently healthy birds, fish aquarium water and other pet animals may pose a health risk to their owners and contacts as all serotypes are known to be potentially pathogenic depending on the oral dosage of the organism and the immune status of those in contact. The high prevalence of resistance to antimicrobial agents among Salmonella isolates across pet species may pose chemotherapeutic consequences to their owners and contacts.     

The pathogenicity of four avian influenza viruses for fowls, turkeys and ducks

Groups of 10 two-week-old chicks, turkey poults and ducklings were each infected by the intranasal route with one of four avian influenza viruses: a/fowl/Germany/34 (Hav 1N))--Rostock, A/FPV/Dutch/27 (Hav 1 Neq 1)--Dutch, A/fowl/Victoria/75 (Hav 1 Neq 1)--Australian, and A/parrot/Ulster/73 (Hav 1 N1)--Ulster. Eight hours after infection 10 birds of the same age and species were placed in contact with each group and allowed to mix. The clinical signs of disease and onset of sickness and death were recorded. Ulster virus was completely avirulent for all birds. Rostock, Dutch and Australian viruses were virulent for fowls and turkeys causing death in all birds with the exception of 3/10 in contact fowls from the Rostock virus group and 2/10 in contact fowls from the Australian virus group. Only Rostock virus caused sicked sickness or death in ducks, 9/10 intranasally infected and 6/7 in contact birds showed clinical signs and 2/10 intranasally infected and 3/7 in contact ducks died. Intranasal and in contact pathogenicity indices were calculated for each virus in each bird species and indicated quantitatively the differences in virulence of the four virus strains. Virus isolation and immune response studies indicated that surviving in contact fowls in the Rostock virus group had never been infected but that surviving Australian virus in contact fowls had recovered from infection. Infection was not established in Ulster virus in contact fowls and Australian virus intranasally infected and in contact ducks. The birds in all other groups showed positive virus isolations and a high incidence of positive immune response. The last virus isolation was made at 22 days after intranasal infection of ducks with Ulster virus.     

Gentamicin and bacterial culture (Nurmi culture) treatments either alone or in combination against experimental Salmonella hadar infection in turkey poults

The effect of three treatments on Salmonella hadar infection in turkey poults was examined. The three treatments were: (A) a single injection of gentamicin, (B) peroral treatment with anaerobic culture derived from cecal contents of an adult turkey (Nurmi culture), and (C) combination of treatments A and B. Nurmi culture was significantly more effective than gentamicin treatment. Nurmi culture along with gentamicin reduced the number of shedding birds, but a higher proportion of birds became carriers compared with the groups given Nurmi culture only. Gentamicin injection alone did not prevent the spreading of infection in larger groups raised on litter, but it did so in one experimental group raised on wire floor.     

Yolk sac, body dimensions and hatchling quality of ducklings, chicks and poults

1. One-day-old chicks of Pekin duck, turkey, layer fowl and broiler fowl were examined for bacteria in the yolk sac and yolk fluid. 2. Whole hatchling, yolk-free hatchling and yolk sac weights were recorded for all species along with crown-rump length and beak-tip to toe-tip length. 3. Bacteriology revealed positive results for the whole yolk sacs of 43 to 64% of the birds in the sample of ducklings, poults and layer chicks. Broiler chicks had a 6.6% incidence of bacteria isolated from the whole yolk sac. By contrast, there were very few positive results from swabs of yolk fluid for any of the bird types. 4. The presence of bacteria in the yolk sac of hatchlings suggests that there is colonisation, rather than infection, of the yolk sac membrane during the hatching period or the first few hours post-hatching. Isolation of bacteria from the yolk sacs of young chicks might no longer be considered as solely indicative of yolk sac infection but further research is required to confirm this result. 5. Contrary to what is being suggested in commercial practice relationships between linear dimensions and hatchling weight suggest that measurement of chick length is at best a very crude measure of chick quality.