Enzyme-linked immunosorbent assay for the detection of Salmonella enteritidis infection in chickens

An ELISA was developed and tested for its ability to detect antibodies against Salmonella enteritidis in chickens. Various features of the ELISA were evaluated and optimized. The outer membrane protein antigens selected by use of the protein immunoblotting method made the assay specific and sensitive. The assay was evaluated in chickens experimentally infected with S enteritidis. Blood samples collected at weekly intervals after experimental infection with S enteritidis were analyzed by ELISA. Results of the ELISA were compared with those of conventional serum plate and microagglutination tests. The ELISA was more sensitive and specific in the detection of S enteritidis infection than the other 2 conventional tests.     

Diagnosis of footrot in goats: application of ELISA tests for response to antigens of Dichelobacter nodosus

Goats are an important natural host for footrot and are infected with Dichelobacter nodosus that have virulence characteristics similar to those of sheep strains. However, the humoral response of goats to D. nodosus antigens and the possibility of a serological diagnosis of footrot in goats have not been studied. With the aim of evaluating a diagnostic ELISA test, we investigated the primary immune response of goats to experimental and natural infection, the memory response in recovered animals, and the transfer and persistence of colostral antibodies in kids. Footrot stimulated the goat's immune system and, as in sheep, under-running lesions were the primary stimulus for production of anti-D. nodosus antibodies. The immune response could be detected in ELISA using either fimbrial or outer membrane protein (KSCN) antigens of D. nodosus. Antibody titres resulting from infection declined quickly after recovery and reached pre-infection levels within 3-4 months. Previously affected animals, however, mounted a memory response when injected with purified D. nodosus antigens. Antibody levels attained after anamnestic challenge were correlated with the maximum levels attained during infection, and were therefore indicative of the infection status. Anti-D. nodosus antibodies were also transferred to kids via colostrum, but these antibodies did not persist and therefore were unlikely to interfere with the diagnostic ELISA after 3 months of age. Though these ELISA tests were highly specific, their sensitivity was rather low. Therefore, they are only suitable for a herd diagnosis of footrot in goats and are dependent on the development of advanced under-running infections in a proportion of affected goats.     

Enzyme immunoassay for surveillance of Q fever

An enzyme-linked immunosorbent assay (ELISA) was developed to monitor antibodies against Coxiella burnetii among animal populations used in research and teaching facilities. Various antigenic components of C burnetii prepared from phase I and phase II whole cells and commercially available antigens were evaluated. A trichloroacetic acid extract was selected for routine use. There was a linear relationship between the transformed absorbance readings of the ELISA results and microagglutination (MA) titers. Comparison between positive or negative results of the MA test and ELISA gave 98.6% concordance. Using the MA test as the standard, ELISA results were 97.8% sensitive and 100% specific. The efficacy of ELISA was evaluated by testing ruminants with known histories of C burnetii infection. Antibody prevalence was 0 in 117 sheep with no history of C burnetii infection, 22% in 145 naturally infected sheep used for research, and 53% in 115 sheep used for vaccine field trials. Forty-eight percent of 120 dairy cows and 52% of 79 goats from endemically infected herds were seropositive. These results indicate that ELISA should be the test of choice for mass screening and surveillance of animals when Q fever is a suspected biohazard.     

Report of a serological study of Coxiella burnetii in domestic animals in Albania

The prevalence of Coxiella burnetii antibodies in domestic ruminants in Albania has been investigated. A total of 1656 serum samples taken from sheep, goats, and cattle housed on farms located in 20 different districts were tested by ELISA for the presence of specific antibodies to C. burnetii phase I and II antigens. Specific IgG antibodies were detected in 9.1% of the animals from both lowland and mountainous areas. In total, a slightly higher percentage of antibodies was detected in sheep and goats (9.8%) than in cattle (7.9%).     

Experimental infection of Red Sokoto goats with Salmonella typhimurium

Salmonella typhimurium infection was experimentally induced in goats by administering 2 x 10(10) organisms per os. The disease produced was characterized by pyrexia, diarrhoea and neutrophilia. One goat died from septicaemia. Somatic "0" agglutinins were detected 14 days post infection (p.i.). Excretion of organisms in faeces ceased by 6 weeks p.i. Goats which recovered from primary infection were refractory to a secondary challenge with 2 x 10(11) organisms. The results indicate that in the absence of signs of ill-health, the detection of neutrophilia and "0" agglutinins and isolation of Salmonella organisms from faeces mainly served as evidence of recent infection.     

Salmonella enteritidis and other Salmonella in laying hens and eggs from flocks with Salmonella in their environment.

Seven Canadian layer flocks with Salmonella enteritidis in their environment were investigated to determine the numbers of hens infected with S. enteritidis, the localization of S. enteritidis in organs of infected hens and the numbers of S. enteritidis-infected eggs produced by two affected flocks. By a microagglutination test (MAT) using S. pullorum antigens, these flocks had more seropositive hens (mean 51.9 +/- 16.9%) than two Salmonella-free flocks (mean 13.0 +/- 4.2%). Culture of tissues of 580 hens (433 seropositive) from the seven flocks detected 26 (4.5%) S. enteritidis-infected hens from two flocks. In one flock, 2/150 hens were infected with S. enteritidis phage type (PT) 8, which was confined to the ceca, and no Salmonella spp. were isolated from 2520 eggs (one day's lay). In the second flock, where 24/150 hens were infected with S. enteritidis PT13, extraintestinal infection was found in nine hens and involved the ovaries and/or oviduct in two hens. Salmonella enteritidis PT13 was isolated from one sample of egg contents and from one sample of cracked shells from among 14,040 eggs (one day's lay) from this flock. The overall prevalence of S. enteritidis-contaminated eggs from the two flocks with infected hens was less than 0.06%. Other Salmonella spp. isolated were S. heidelberg from 58 hens (10%), and S. hadar, S. mbandaka and S. typhimurium from one hen (0.2%) each. The MAT with antigens of S. pullorum had a sensitivity of 81% and a specificity of 24% for detecting S. enteritidis-infected hens.(ABSTRACT TRUNCATED AT 250 WORDS)     

Corynebacterium pseudotuberculosis infection in goats. III. The influence of age

Serological examinations for Gorynebacterium pseudotuberculosis in-fection were carried out in 14 known positive herds and in 1 herd that had beeni recently established through purchase of animals from different herds. Serum samples were collected sequentially on up to 4 occasions from animals during their first year of life. In most herds, these animals were examined clinically for superficial swellings once or twice during the same period.The adult goats in 8 infected herds were examined both clinically and serologically. Age distribution was similar in each of these herds. All serum samples were examined for antibodies against Gorynebac-terium pseudotuberculosis in both the bacterial agglutination test (BAT) and the hemolysis inhibition test (HIT).The proportion of kids which were seropositive in both tests de-creased to zero at 4 months of age. At the age of 11–12 months, the proportion of seropositive animals was about 50 % in the BAT and 30 % in the HIT.When examined when housed for the winter at the age of about 8 months (mean age), the prevalence of animals with superficial swellings was 7 %. At the age of about 1 year (mean age), 29 % of the examined animals had such lesions. In the recently established herd, only a few of the yearlings were seropositive.Titre values in BAT and HIT increased until 6 years of age. Anti-body titre values were significantly lower in yearlings than in older goats in both tests, P < 0.005. No significant difference in the propor-tion of goats with superficial swellings was seen at the different ages.     

Survey of clinical psittacine bird sera for Salmonella typhimurium agglutinins.

Of 2407 serum samples from various kinds of psittacine birds submitted for Chlamydia serology, 2343 (97.4%) were negative, 25 (1.0%) were equivocal, and 39 (1.6%) were positive for Salmonella typhimurium agglutinins. In additional serum samples from two groups of African gray parrots, the prevalence of agglutinins was 0.0% (0/38) in the Timneh variety and 24.0% (6/25) in the Congo variety. In sera from one macaw, one cockatoo, and one Amazon parrot, which were negative for chlamydial antibody activity, there were strongly reactive agglutinins for S. typhimurium. Two Amazon parrots had antibody activity against Salmonella and Chlamydia antigens.     

THE ENZYME-LINKED IMMUNOSORBENT ASSAY (ELISA) AS A SEROLOGICAL TEST FOR DETECTING ANTIBODIES AGAINST LEPTOSPIRA INTERROGANS SEROVAR HARDJO IN SHEEP

SUMMARY: The enzyme-linked immunosorbent assay (ELISA) was compared with the standard microscopic agglutination test (MAT) as a method for detecting antibodies against Leptospira interrogans serovar hardjo in sheep. Peak antibody levels detected by the 2 tests occurred at different times following experimental infection of sheep. In serums from flocks of sheep with naturally acquired infection there was a 95% correlation between MAT and ELISA with respect to the presence or absence of antibody to serovar hardjo , although the level of correlation of the titres of the 2 tests was low. The 2 tests appeared to measure different antigen-antibody systems. The ELISA would be a useful test for screening large numbers of serums for antibodies to L. interrogans serovar hardjo .     

Anti-Toxocara spp. antibodies in sheep from southeastern Brazil

The purpose of this study was to evaluate the frequency of anti-Toxocara antibodies in sheep from Presidente Prudente, southeastern Brazil. Serum samples were obtained from 365 sheep of diverse breeds and different ages. Samples were collected at a slaughterhouse and at farms located in Presidente Prudente. Three groups of animal of different ages were evaluated according to age: Group I: between 1 and 6 months old; Group II: between 7 and 10 months old; and Group III: between 11 and 15 months old. An ELISA test was carried out to detect anti-Toxocara antibodies (IgG) using the excretory-secretory antigens of Toxocara canis (TES) larvae. In total, 183 out of 365 animals (50.1%) were positive for anti-Toxocara antibodies. The frequency of antibody detection was directly proportional to the age of the animals (p<0.0001), indicating a relationship between infection and aging. In Group III, there was a higher prevalence in females (p=0.0041). The relevance of these animals to the epidemiology of toxocariasis in pets and human should be considered.     

A comparison of four serological tests for the diagnosis of caseous lymphadenitis in sheep and goats

A double antibody sandwich ELISA (ELISA A) developed for the detection of Corynebacterium pseudotuberculosis infection in sheep and goats was modified to improve its sensitivity. To establish the sensitivity and specificity of this modified ELISA (ELISA B), sera from 183 sheep and 186 goats were tested using ELISAs A and B. Comparison was also made with two further ELISAs (C and D) developed in Australia that, respectively, detect antibodies to cell wall antigens or toxin.ELISA B had the best performance of the four tests. Its specificity was 98+/-1% for goats and 99+/-1% sheep. Its sensitivity was 94+/-3% for goats and 79+/-5% for sheep. ELISA B will now be tested for use in caseous lymphadenitis eradication and control programmes in The Netherlands. It will also be used in experimental studies of CL in Scotland.     

Seasonal variation of Oestrus ovis-specific antibodies in sheep and goats mixed flocks in Greece

The aim of this survey was to investigate the year-round epidemiological patterns of Oestrus ovis ELISA sero-prevalence in sheep and goats kept together under the same husbandry system in an endemic area of Greece. Twenty-five adult female sheep and 25 adult female goats, coming from a large mixed flock, were randomly selected, eartaged and monthly blood sampled during 1 year period (November 1998-October 1999). Serological prevalence in sheep was 100% all around the year. Mean intensities of specific O. ovis antibodies follow a seasonal evolution with higher mean titers between March and July than in winter. In contrast, the serological prevalences in goats were low specially in winter months (from October to January). No significant difference were noticed in goats antibody levels during the year period. The possible reasons of this difference of O. ovis sero-prevalence between sheep and goats are discussed.     

Antibody seroprevalences against Peste des Petits Ruminants (PPR) virus in sheep and goats in Sudan

Counter immnuo-electrophoresis (CIEP) and Competitive ELISA (C-ELISA) tests were employed for seroprevalence of Peste des Petits Ruminants (PPR) infection in Sudan. The result of both tests showed high prevalence of PPRV antibodies in sheep and goats sera collected from six different regions of Sudan. Of the 519 serum samples examined for the presence of PPRV antibodies 307(59.15%) were positive by CIEP while 263(50.67%) were positive by C-ELISA. CIEP technique was shown to be more sensitive than C-ELISA technique for detection of PPRV antibodies (Kappa statistics 0.259). C-ELISA allowed rapid, simple, specific, sensitive and differential sero-diagnosis of PPRV and RPV in sheep, goats and cattle. CIEP is, unlike competitive ELISA, is group-specific test and can not differentiate between PPR and RP infections. Despite its low specificity CIEP can be a useful indicative screening test for PPRV antibodies in flocks that neither been vaccinated nor otherwise exposed to PPR or RP virus. Results obtained suggest that CIEP, like the HI test, could be a useful screening test where it is not possible to use C-ELISA.     

Prevalence of antibodies against Babesia canis in dogs in an endemic area

A survey of 287 dogs for antibodies against Babesia canis in dogs in an endemic area, using ELISA, produced a prevalence of 43 per cent. Antibodies occurred in dogs of all age groups, the prevalence being significantly lower in dogs aged 1 to 6 months than in older dogs. There were no differences between indigenous Nigerian dogs and exotic (foreign) dogs; and between the sexes in the prevalence of antibodies. Antibodies were more prevalent in dogs with B. canis parasitaemia and in those with a higher risk of infection. Also antibodies were detected in some puppies born to seropositive bitches. The ELISA test failed to detect antibodies in 36.1 per cent of dogs with B. canis parasitaemia.     

Seroprevalence and risk factors of caprine toxoplasmosis in Minas Gerais, Brazil

The objective of this work was to carry out a study on caprine toxoplasmosis in the state of Minas Gerais, Brazil. To determine the prevalence of toxoplasmosis in goats in Minas Gerais, 767 sera from goats were tested by ELISA (enzyme-linked immunosorbent assay) and IFAT (indirect fluorescence antibody test). The prevalence of antibodies to Toxoplasma gondii was 43.0% and 46.0% by ELISA and IFAT, respectively. It was observed that 26.8% of the goats show low-avidity IgG to T. gondii. These results suggest the presence of animals in recent phase of toxoplasmosis in Minas Gerais. The risk factors for toxoplasmosis in goats were: age over 36 months (OR = 1.21; IC 95% 1.02–1.44), use of pen (OR = 1.83; IC 95%1.01–3.31) and pure breed animals (OR = 2.49; IC 95% 1.11–5.59).     

Prevalence and mortality rate of peste des petitis ruminant (ppr): possible association with abortion in goat

Present study was designed to investigate the prevalence and mortality (%) caused by Peste des Petitis Ruminant (PPR) and its possible association with abortion in goat flocks at different areas of Pakistan. A total of 140 animals were samples in the population of 650 which was having 185 deaths (Mortality rate = 28 %) from three different regions of the country. There were 58 abortions in the 140 pregnant goats of above said population One hundred & ten (110) serum samples from diseased, recovered and apparently healthy animals were tested for the presence of PPR antibodies by competitive ELISA (c ELISA). Eighty-four (84) animals were positive for PPR antibodies whereas in apparently healthy adult goats in the same flock, no PPR antibodies were detected. Twenty-four (24) tissue samples collected from the dead animals and six samples from aborted fetus were tested for the presence of PPR antigen by Immuno-capture ELISA (Ic ELISA). Nineteen (19) out of thirty (30) organ samples mainly from lung, spleen, lymph node were found positive for PPR antigen but negative from lungs of aborted fetus. There was a high rate of abortions (28–45 %) in each of the outbreak and it was highest in the outbreak of Golra Sharif, Islamabad (No. = 21 in total population of 100). As the serum samples from the aborted dams were found positive for PPR antibodies so the study provides the possible association of mortality and prevalence of PPR disease with high rate of abortions in goat.     

Serological detection of experimental Salmonella enteritidis infections in laying hens

The antibody response of laying hens to experimental Salmonella enteritidis infection was evaluated in microagglutination, tube agglutination, and rapid whole-blood plate agglutination assays. Hens of three different ages were infected by either oral inoculation or horizontal contact transmission. Blood was collected at weekly intervals, and the presence of specific antibodies was assessed by reaction with antigens prepared from strains of S. enteritidis and S. pullorum. The sensitivity of detection of infected hens did not vary significantly between the assays, as all three tests effectively identified most exposed hens as seropositive. Within each test, however, variation was observed in the detection sensitivity when different antigens were used. The microagglutination titers of serum samples were determined by serial dilution. Antibody titers peaked at 1 to 2 weeks postinoculation and declined steadily, although most birds were still identified as seropositive at 10 weeks postinoculation. The mean microtest titers obtained with S. enteritidis antigens were higher than with an S. pullorum antigen, indicating greater test sensitivity. However, use of the S. pullorum antigen resulted in fewer false positives when sera from uninfected control hens were tested. The titers of contact-exposed hens peaked later and at lower values than did those of inoculated hens, but these two groups of hens had similar antibody titers after the third week postinoculation.     

Assessing the agreement between Ostertagia ostertagi ELISA tests performed using the crude adult antigen and the adult and larval stage 4 excretory/secretory antigens

The objective of this study was to determine the agreement between ELISA tests conducted using three O. ostertagia antigens: crude adult worm, larval stage 4 (L4) excretory/secretory (ES) and adult ES. This study was carried out on 289 Holstein cows from five herds in Prince Edward Island and one herd in Nova Scotia. Composite milk samples of these cows were collected (between May and September 2002) from the respective provincial laboratories and sent to the Atlantic Veterinary College where each sample was tested for antibodies to O. Ostertagi using an indirect microtitre ELISA test. Results were expressed as optical density ratio (ODR) values. Each milk sample was tested with three ELISA tests, with each test using a different O. ostertagi antigen. There was a slight rise in ODR values of both adult antigens, between May and August, with higher values obtained using the adult ES antigen. L4 ES ODR values were generally higher than those for both adult antigens during the study period, except for May. There was a more dramatic rise in L4 ES ODR values between May and August. Rises in ODR in May and end of July coincided with periods of mass maturation of L4 to adult worms. The results of the study showed that the concordance correlation coefficient (CCC) between tests performed using both ES and the crude antigens were low (crude adult versus adult ES=0.31, crude adult versus L4 ES=0.30). The highest CCC was observed between tests done using both ES antigens (CCC=0.56). Generally, the study results suggest that the antibody response (detectable by the ELISA) is mainly directed against ES antigens (especially L4) than the crude adult worm antigen.     

Prevalence of Bovine viral diarrhoea virus (BVDV) antibodies among sheep and goats in India

The aim of this study was to determine the prevalence of pestivirus antibodies in sheep and goats in India. A total of 2803 serum samples collected between 2004 and 2008 from 1777 sheep in 92 flocks and 1026 goats in 63 flocks belonging to 13 states were tested by competition ELISA for detection of pestivirus antibodies. In sheep, the true prevalence rate was 23.4% (95% confidence interval: 22.9%–27.0%) and in goats it was 16.9% (95% CI: 16.4%–21.3%). The flock level seroprevalence was 66.3% for sheep and 54.0% for goats. Geographical variation in individual and flock prevalence was highly significant. A significant association (p?<?0.05) was found between sheep and goat flocks having cattle contact and the flock level seroprevalence. The seroprevalence was lower in 6 months–1 year age group compared to the 1–2 year and >2 year age groups in both sheep and goats. Cross neutralization studies on 61 seropositive sheep and 34 seropositive goat samples representing all positive flocks, exhibited > four fold higher titre to bovine viral diarrhoea virus type 1 (BVDV-1) in 41 sheep and 23 goat samples and to BVDV-2 in one sheep and goat each. This study for the first time showed serological evidence of wide spread BVDV infections in Indian sheep and goats, with BVDV-1 predominating and BVDV-2 occasionally besides highlighting the potential risk of infection to other species, which needs to be considered whenever BVD control measures are initiated.     

Helminths of goats in Mongolia

Post-mortem examinations of 236 goats from all provinces in Mongolia were performed for the study of helminths in goats. Thirty-nine helminth species belonging to three classes, 14 families and 23 genera were found. Trichocephalus spiralis and Avitellina centripunctata are reported for the first time in goats in Mongolia. The prevalence and intensity of helminth infections are reported for three age groups of goats in four seasons and three geographic zones in Mongolia. Common helminth infections of goats in all zones of Mongolia were infections of Ostertagia, Marshallagia and Nematodirus. In addition, fecal samples of 15 kids, 15 yearlings and 15 adult goats were examined monthly for eggs. The highest number of eggs per gram (EPG) of feces was counted in March (average 1335.3+/-405.3) and the lowest count was in November (54+/-18.6).