Campylobacter fetus subsp. jejuni in poultry reared under different management systems in Nigeria

Cloacal swabs from 487 live birds in 36 flocks and 70 poultry carcasses were cultured for Campylobacter fetus subsp. jejuni. It was isolated from 12.3% of the birds in 19 flocks. Chickens, turkeys, and guinea fowl differed from one another in isolation rates of the organism. Management system affected its occurrence, and only 7.1% of eviscerated carcasses yielded it. It was concluded that bird species, management system, and immersing slaughtered poultry in boiling water before dressing affect recovery of C. fetus subsp. jejuni from live birds and carcasses.     

Occurrence and significance of different Campylobacter types in domestic animals in Hungary

Experiences, including results of original experimental work on Campylobacter fetus, C. jejuni and C. coli induced diseases of cattle, sheep, dogs, rabbits poultry and men in Hungary are reviewed. Out of 31 cases of abortion in cows 29 (93.5%) were causes by C. fetus subsp. venerealis and only one case each (3.2%) by C. fetus subsp. fetus and C. jejuni, respectively. Out of the 29 strains of C. fetus subsp. venerealis, 26 belonged to serogroup 01 (A) and only 3 to serogroup 02 (B). Campylobacter abortions in sheep flocks were caused in 18 cases (78.3%) by C. fetus subsp. fetus and in 5 cases (21.7%) by C. jejuni. The latter strains belonged to Penner's serogroup 1 (6 strains), 5 (4 strains) and 8 (5 strains), respectively. In scouring dogs 12.7% of the cases were caused by C. jejuni. The same pathogen caused diarrhoea also in young rabbits. Isolated strains belonged to serogroup 2. In cases of Campylobacter hepatitis of laying hens, egg production has been reduced by 8 to 15% for 2 to 3 weeks. Row poultry meat represents often the source of infection for men. The 32 strains of C. jejuni isolated from faecal samples of men affected with diarrhoea belonged to 12 serogroups.     

Serogroups of Campylobacter fetus and Campylobacter jejuni isolated in cases of ovine abortion

Of 38 aborted ovine fetuses from 23 sheep flocks 29 C. fetus subsp. fetus and 22 C. jejuni were isolated and examined biochemically and serologically for heat-stable antigens. Serologic examinations were carried out by passive haemagglutination test. In case of C. fetus subsp. fetus strains alkaline antigen extraction was used. Antisera to two serogroups of C. fetus and to Penner serotype reference strains 1 to 60 were produced in rabbits. Abortion was caused in 18 (78.3%) flocks by C. fetus subsp. fetus and in 5 (21.7%) flocks by C. jejuni. Six C. fetus subsp. fetus strains grew well at both 43 and 25 degrees C. With one exception all C. fetus subsp. fetus were resistant, whereas all 29 C. fetus subsp. venerealis strains were sensitive to 30 micrograms/ml cefoxitin and cefamandole. These two cephalosporins can be used to differentiate the two subspecies of C. fetus. Passive haemagglutination test using alkaline antigen extraction is a proper method for the examination of heat-stable antigens of both C. fetus subspecies. Out of 24 C. fetus subsp. fetus strains 13 belonged to serogroup A(01), and 11 to serogroup B(02). C. jejuni strains examined belonged to Penner serogroup 1 (6 strains), to serogroup 5 (4 strains) and to serogroup 8 (4 strains).     

Pulsed-field gel electrophoresis typing of Campylobacter fetus subsp. fetus from sheep abortions in the Hawke's Bay region of New Zealand

AIM: To type Campylobacter isolates from sheep abortions from the Hawke's Bay region of New Zealand. METHODS: Campylobacter isolates were collected from aborted sheep fetuses from commercial farms in the Hawke's Bay region. Information on the Campylobacter vaccination status of flocks in the study was collected. Isolates were identified to species level using standard phenotypic tests, then typed using pulsed-field gel electrophoresis (PFGE). RESULTS: Eighty-one C. fetus subsp. fetus isolates were cultured from aborted sheep fetuses from 25 farms and four C. jejuni isolates were cultured from fetuses from three farms. The C. fetus subsp. fetus isolates were classified into six PFGE groups. A single pulsed-field type predominated amongst isolates from 19 of the 25 farms. The C. jejuni isolates comprised two types. CONCLUSIONS: A range of C. fetus subsp. fetus PFGE types was identified, and one type, B1, was found most frequently. Campylobacter fetus subsp. fetus was only isolated from samples from sheep that had not been vaccinated with C. fetus subsp. fetus vaccine that season.     

Campylobacter jejuni abortion in a heifer

Campylobacter jejuni was isolated from the lung and stomach contents of an aborted bovine fetus. The bacterial isolate was classified according to morphologic, biochemical, and thermotolerance characteristics as well as sensitivity to antibiotics and biochemical agents. Serotyping was specific for C jejuni and excluded this isolate from the more common bovine causes of campylobacteriosis, C fetus subsp venerealis or C fetus subsp fetus. This report stresses the need for careful speciation of all Campylobacter isolates from aborted fetuses, especially in conditions where routine vaccination has not reduced abortion rates in the herd. Campylobacter jejuni is much more prevalent in causing human infections than is C fetus subsp venerealis or C fetus subsp fetus. Therefore, from a public health standpoint identification is important.     

Bulk growth procedures and a button agglutination test for Campylobacter

Bulk-cell yields were obtained from 4 Campylobacter spp incubated aerobically without the use of a special atmosphere. A button agglutination test was developed for quantitation of blood serum antibodies against C fetus subsp venerealis, C fetus subsp fetus, C jejuni, and "C hyointestinalis." The test was sensitive, and different individuals reading it usually attained the same titers. Cells of C fetus subsp venerealis, C fetus subsp fetus, and "C hyointestinalis" grown aerobically in broth made satisfactory antigens for the button test, but some cell pools of C jejuni had a tendency to autoagglutinate. Cells of C jejuni grown on blood agar had less tendency to autoagglutinate.     

Adaption of ELISA for the detection of Campylobacter antibodies and its application in seroepidemiological studies in sheep and cattle herds

ELISA was adapted for the study of antigenic relations among important Campylobacters and for the presence of anti-campylo-bacter antibodies in 394 sheep and 265 cattle. Rabbit anti-C. jejuni, C. coli, G. fetus subsp. fetus and C. laridis heat-stable antigen sera were evaluated against 29 Campylobacter strains and 6 other bacteria. Anti-C. jejuni and G. coli reacted strongly with homologous antigens and weakly with C. fetus subsp. fetus, C. laridis and C. fecalis antigens. C. fetus subsp. fetus serum reacted mainly with its homologous antigen. C. laridis serum showed closer reactivity to C. jejuni than to C. fetus subsp. fetus, C. coli and C. fecalis. Insignificant cross-reactions were observed with Y. enterocolitica, S. dublin and E. aerogenes heat-stable antigens, Ewes vaccinated with C. fetus subsp. fetus bacterin showed higher ELISA titers against C. fetus subsp. fetus antigens than non-vaccinated ewes or rams. Twenty-five percent of the vaccinated animals showed titers as low as 95 % of the non-vaccinated animals. In cattle the lowest antibody titers against C. fetus subsp. fetus, C. jejuni, C. coli and C. laridis antigens were exhibited by the precolostrum sera followed by the postcolostrum and adult sera. These studies demonstrated the applicability of the ELISA test in seroepidemiological investigations concerning the distribution and significance of Campylobacter antibodies in food animal sera.     

Campylobacter as the cause of diarrhea in calves]

The modified Preston medium allows the isolation of C. jejuni, C. coli and C. fetus subsp. fetus from intestinal samples of calves at an incubation temperature of 37 degrees C. In the first series of investigation, Campylobacter excretion in calves (n = 7) was followed up to the age of 4 months. In the first 4 days of life, these bacteria could not be detected in any of the animals. Thereafter first C. coli we found in all calves. In 4 animals, only strains of this species were isolated during the whole investigation period. In 3 animals C. fetus subsp. fetus could be detected repeatedly, however C. coli and sometimes C. jejuni were found, too. In the second series of investigation, isolation of Campylobacter from different parts of the gastrointestinal tract or organs was successful in 19 out of 25 diarrhoeal, moribund calves. 16 out of 19 positive animals harboured large amounts of these gramnegative bacteria in the distal jejunum and ileum. In 10 animals out of these 16, the germ colonized also the proximal jejunum and abomasum. From 6 calves, C. fetus subsp. fetus was isolated, and C. jejuni from 7 calves. C. coli was relatively rare. From the lymph nodes of the proximal and distal jejunum, Campylobacter (exclusively C. jejuni) were isolated from 5 animals. Due to the Campylobacter presence in the small intestine of diarrhoeal calves, a contribution of this bacteria within the pathogenesis of calf diarrhoea is possible. Final evaluation of their pathogenesis importance is only positive by means of virulence tests.     

Pulsed-field gel electrophoresis of Campylobacter jejuni sheep abortion isolates

Campylobacter species are a significant cause of sheep abortion in most sheep-raising countries. In New Zealand, Campylobacter fetus subsp. fetus is the leading cause of diagnosed sheep abortion and the species C. jejuni and C. coli have also been implicated. To date, strain typing information of C. jejuni sheep abortion isolates is limited. The objective of the present study was to genotype C. jejuni isolates cultured from sheep abortions submitted to diagnostic laboratories in New Zealand during the 2000 breeding season, using pulsed-field gel electrophoresis (PFGE). In this study, C. jejuni isolates were cultured from approximately 10% of farms from which Campylobacter species were isolated from sheep abortions in the year 2000. This equated to 25 C. jejuni isolates from 21 farms. These isolates were obtained from the veterinary diagnostic laboratories and strain typed using the molecular typing technique PFGE. Ten distinct PFGE types were identified amongst the isolates. No particular PFGE type was found most frequently amongst these C. jejuni sheep abortion isolates. However, indistinguishable or similar C. jejuni PFGE types were identified from different aborted foetuses from the same flock, consistent with the role of C. jejuni as an infectious cause of abortion in sheep. These strain types were similar or indistinguishable from C. jejuni sheep abortion isolates obtained in 1999 in a smaller study (Mannering, S.A., Marchant, R.M., Middelberg, A., Perkins, N.R., West, D.M., Fenwick, S.G., 2003. Pulsed-field gel electrophoresis typing of C. fetus subsp. fetus from sheep abortions in the Hawke's Bay region of New Zealand. NZ Vet. J. 51, 33-37).     

Comparative studies on competitive exclusion of three isolates of Campylobacter fetus subsp. jejuni in chickens by native gut microflora

Resistance of young chicks to Campylobacter fetus subsp. jejuni was substantially increased by early exposure to native gut microflora. Protection was demonstrated against two human isolates and a chicken isolate of C. fetus subsp. jejuni. Significant protection against the chicken isolate was observed throughout a 91-day test period. Infection reached 100% (25/25) in the untreated group at 56 days of age and only 4% (1/25) in the group treated with native gut microflora. Campylobacter fetus subsp. jejuni was isolated from the ceca and less frequently from the gall bladder and liver of chicks that actively shed the bacteria. Cultures of feces from chicks reared on wood-shavings litter were often negative, suggesting that culturing litter as an indicator of infection has limited value.     

The prevalence and genetic diversity of Campylobacter spp. in domestic 'backyard' poultry in Canterbury, New Zealand

Campylobacteriosis is the most commonly notified illness in New Zealand. Whilst the importance of commercial poultry in campylobacteriosis is well established, little is known about the possible role of chickens kept at home as a direct animal/faecal contact or consumption exposure pathway. The aim of this study was to determine the prevalence and genetic diversity of Campylobacter spp. in domestic backyard chicken flocks in the Canterbury region of New Zealand. Poultry faecal samples were collected from 35 domestic 'backyard' poultry flocks from urban and rural properties around the Canterbury Region of New Zealand. A total of 291 samples were collected and tested for the presence of thermotolerant Campylobacter spp. and positive isolates were analysed using pulsed-field gel electrophoresis (PFGE) using both SmaI and KpnI enzymes. There was a high prevalence of Campylobacter spp. with 86% of flocks testing positive. Campylobacter jejuni alone, Campylobacter coli alone and both C. jejuni and C. coli were detected in 20 (57%), 2 (6%) and 8 (23%) of the flocks respectively. SmaI/KpnI PFGE analysis identified 50 different genotypes across the 35 flocks. Genotype diversity richness was highest on the lifestyle block and farm properties with 43 different genotypes isolated, whilst urban properties displayed the least richness with 12 genotypes isolated. Rural flocks tended to have more different genotypes in a given flock than urban flocks. Comparison of the genotypes with the PulseNet Aotearoa Campylobacter database showed that 28 of the genotypes had previously been isolated from human cases of campylobacteriosis. Many of these were also indistinguishable from Campylobacter spp. previously isolated from retail chicken. Therefore, contact with backyard poultry or their faecal material is a potential additional infection pathway outside of exposure to the established pathways associated with the consumption of Campylobacter-contaminated commercial meat or foods cross-contaminated from contaminated poultry.     

A DUAL VACCINE FOR THE IMMUNISATION OF CATTLE AGAINST VIBRIOSIS

Single subcutaneous injections of a mineral oil adjuvant vaccine containing 20 mg dry weight of Campylobacter fetus subsp fetus biotype venerealis cells and 20 mg dry weight of C. fetus subsp fetus biotype intermedius cells per 5 mil dose protected 2- and 3-year-old heifers and 3- and 4-year-old cows against genital infection with either organism.     

Prevalence of Campylobacter jejuni and its resistance to antibiotics in poultry in the Czech Republic

Thermotolerant Campylobacter spp., in particular Campylobacter jejuni, are among the most frequently identified pathogens, found to be causing human gastrointestinal infections in Europe, with the Czech Republic being no exception. The presented work aimed at assessing results of the first nationwide monitoring of prevalence and antibiotic resistance of Campylobacter spp. in broiler flocks in the Czech Republic, including a comparison of antibiotic resistance of C. jejuni isolates collected from poultry and the human community. The monitoring was carried out in poultry slaughterhouses in 2006 and 2007. From broilers, cloacal swabs were collected and examined. The human isolates of C. jejuni were acquired from rectal swabs in community patients with diarrhoeal diseases. Suspected isolates of both animal and human origin were confirmed by the PCR methods. Antibiotic resistance to selected anti-microbial agents was tested by the microdilution method. In the monitored period, the prevalence of thermotolerant Campylobacter spp. in broilers in the Czech Republic reached almost 50%. In 2006, C. jejuni was detected in 46% and Campylobacter coli in 3% of the tested samples. In 2007, C. jejuni was found in 43% and C. coli in 2% of the samples. The results of anti-microbial susceptibility testing of C. jejuni showed higher resistance in animals when compared with humans. The only exception was tetracycline with higher resistance in isolates of human origin. The highest resistance detected was to quinolone antibiotics. Resistance to oxolinic acid was 77% in animal and 60% in human isolates, to ciprofloxacin 72% in isolates from poultry and 55% in those from humans. In ampicillin, 26% of poultry isolates and 16% of human isolates were resistant. Moreover, 9% of animal isolates demonstrated resistance to streptomycin, undetected in human isolates. In erythromycin, resistance was found in 6% of poultry and 1% of human isolates.     

Epidemiology of ovine Campylobacter infection determined by numerical analysis of electrophoretic protein profiles

The relationship of 50 Campylobacter strains isolated from aborted ovine foetuses, and the faeces of sheep, cattle and chickens were determined by numerical analysis of electrophoretic (SDS-PAGE) protein profiles. Comparison of protein patterns by numerical methods revealed differences between C. fetus ssp. fetus, C. jejuni, and C. coli strains as well as heterogeneity among isolates from different outbreaks. Isolates from each farm produced a distinct cluster and flocks from different locations were found to be infected with relatively different strains. In most cases, protein patterns of ovine foetal isolates were very similar to those of ovine faecal isolates. Ovine isolates of C. fetus ssp. fetus, C. jejuni and C. coli gave similar protein patterns to the corresponding Campylobacter species isolated from cattle or chicken, on the same farm. Thus, it was concluded that certain protein types of ovine Campylobacter strains were more likely associated with local areas, and Campylobacter strains causing ovine abortions are distributed in the environment more widely than assumed to date.     

Campylobacter colonization in poultry: sources of infection and modes of transmission

Since its recognition as a human pathogen in the early 1970s, Campylobacter jejuni has now emerged as the leading bacterial cause of food-borne gastroenteritis in developed countries. Poultry, particularly chickens, account for the majority of human infections caused by Campylobacter. Reduction or elimination of this pathogen in the poultry reservoir is an essential step in minimizing the public health problem; however, farm-based intervention measures are still not available because of the lack of understanding of the ecological aspects of C. jejuni on poultry farms. Although Campylobacter is highly prevalent in poultry production systems, how poultry flocks become infected with this organism is still unknown. Many investigations indicate that horizontal transmission from environmental sources is the primary route of flock infections by Campylobacter. However, some recent studies also suggest the possibility of vertical transmission from breeder to progeny flocks. The transmission of the organism is not well understood, but it is likely to be through both vertical and horizontal transmission and may be affected by the immune status of the poultry host and the environmental conditions in the production system. Intervention strategies for Campylobacter infection in poultry should consider the complex nature of its transmission and may require the use of multiple approaches that target different segments of the poultry production system.     

Serotype and genotype diversity and hatchery transmission of Campylobacter jejuni in commercial poultry flocks.

We investigated the genotype and serotype diversity of Campylobacter coli and C. jejuni in two parent flocks of adult hens and their offspring over two rotations in order to evaluate the role of hatchery mediated transmission and/or vertical transmission of campylobacters in broiler flocks. In total, 314 C. jejuni and 32 C. coli isolates from parent and broiler flocks and from the surroundings of broiler houses were typed by flagellin gene PCR/RFLP (fla-typing), and selected isolates were also typed by serotyping and macrorestriction profiling using PFGE (MRP/PFGE). The combined typing results showed that the broiler flocks could be colonised by 1-3 different Campylobacter clones and parent flocks could be colonised by 2-6 different clones. C. coli was isolated from up to 36% of birds in one parent flock, whereas only C. jejuni was isolated from broiler flocks. C. jejuni clones from different flocks were clearly discriminated by fla-typing as well as by MRP/PFGE, except for a few cases where individual isolates belonging to two different clones were found to have altered fla-types. Similarly, one C. coli clone showed pronounced fla-type variation. The present results lead to the conclusion that vertical transmission or horizontal transmission via the hatchery are not significant transmission routes of C. jejuni to broiler chickens under Danish conditions. In the cases where more than one Campylobacter clone simultaneously colonised flocks, we found that the different clones coexisted in flocks rather than excluding each other.     

Pathogenicity of Campylobacter jejuni and Campylobacter coli strains in the pregnant guinea pig model

Pathogenicity of 17 Campylobacter isolates for pregnant guinea pigs was investigated. Of 14 isolates, 12 (86%) produced rates of abortion ranging from 13% to 87%. Two isolates did not produce abortion. Reference strains of C fetus subsp venerealis produced abortion in 60% to 87% and C fetus subsp fetus produced abortion in 60% of the guinea pigs. Inoculated organisms were recovered from uterus, blood, liver, kidney, spleen, and gallbladder of the guinea pigs at rates as high as 83% for 2 ovine isolates and as low as 13% for 2 bovine and 1 human isolates. Most isolations were from the uterus. Two avian isolates were not recovered. Within the C jejuni and C coli group, the ovine and the human isolates appear to be more pathogenic. Swine, bovine, and avian isolates were less pathogenic. Seemingly, the pregnant guinea pig was a suitable and practical model for evaluating the pathogenicity of Campylobacter organisms, regardless of their host of origin.     

Investigation of bovine venereal campyloacteriosis in beef cow herds in New Zealand

AIMS: To determine regional prevalences of beef cow herds in New Zealand positive for Campylobacter fetus subsp venerealis antibodies in samples of vaginal mucus tested using an immunoglobulin (Ig) A enzyme-linked immunosorbent assay (ELISA), and to examine the suitability of the IgA ELISA for detecting infection with C. fetus subsp venerealis under field conditions in New Zealand. METHODS: Vaginal mucus samples (n=1,230) collected from beef cow herds (n=125) throughout New Zealand (approximately 10 samples/herd) were tested for antibodies to C. fetus subsp venerealis using an IgA ELISA. Test results were compared between herds classified as having low, medium and high fertility based on pregnancy test results interpreted in relation to the duration of the mating period used. In addition, a small number of samples were collected from dairy cows that were mated using artificial insemination (AI) and had no contact with breeding bulls. The influence of putative risk factors for the spread of venereal disease and the effect of sample quality on the status of herds according to test results was assessed using multivariate logistical regression. Preputial washings from 54 bulls from nine herds classified as low fertility in which mucus samples from > or =3 cows were IgA antibody-positive were cultured for the presence of Campylobacter spp, and isolates of C. fetus subspecies were characterised using a polymerase chain reaction (PCR) test. RESULTS: One or more mucus samples was positive to the IgA ELISA in 70% of all herds tested. The prevalence of IgA antibody- positive individuals was >20% in most regions of New Zealand and did not differ significantly for cows from herds classified as high, medium or low fertility (28%, 26% and 23%, respectively; p=0.39). No relationship was found between mucus antibody status and age of breeding group, herd size, herd fertility, number of herds that female replacements or breeding bulls were sourced from, whether a serving ability test (SAT) was used to assess bulls, or the quality of samples submitted to the laboratory. Campylobacter fetus subsp venerealis was not cultured from any of the 54 bulls sampled. Four other species of Campylobacter and related organisms were cultured, viz Arcobacter cryaerophilus, Campylobacter jejuni, Campylobacter fetus subsp fetus and Helicobacter cinaedi. CONCLUSIONS: The specificity of the IgA ELISA as a diagnostic test for C. fetus subsp venerealis was found to be unsatisfactory under New Zealand conditions. It is possible that an immunological response by cows to Campylobacter species other than C. fetus subsp venerealis caused cross-reactivity in the IgA ELISA. The results do not support the hypothesis that C. fetus subsp venerealis is widespread in New Zealand.     

Potential use of characterised hyper-colonising strain(s) of Campylobacter jejuni to reduce circulation of environmental strains in commercial poultry

Sixty-two persistently colonising Campylobacter jejuni strains were tested for their ability to dominate colonisation of the chicken gastrointestinal tract in competition with each other leading to selection of dominant or "hyper-colonising"Campylobacter strains, which are able to displace others in the chicken intestinal tract. One such strain was shown to be a hyper-efficient coloniser of chickens, as it was able to displace other colonising strains, as well as maintain itself in the chicken intestinal tract for the duration of the 56-day broiler production cycle. Once colonisation was established, this hyper-colonising C. jejuni strain, 331, could not be displaced by other colonising or hyper-colonising strains. We proposed that a defined, hyper-colonising strain, or a cocktail of defined strains with a similar phenotype, could form the basis for biological control of unknown/uncharacterised Campylobacter strains from the environment that continuously colonise chicken flocks. To validate this approach, three different chicken infection trials were carried out. These trials demonstrated that the dominant strain of C. jejuni was able to colonise broiler chickens consistently and for the entire life of the birds irrespective of the day of inoculation and antimicrobial agents used in the feed to control other pathogenic micro-organisms. In addition, we have shown that the bio-control strain was able to replace other colonising strains at various points of a 56-day broiler production cycle irrespective of time and type of inoculation. This strain was also capable of re-establishing itself following the challenge with other strains, with and without re-challenge. This work represents a "proof of principle" that a defined C. jejuni strain could be used to biologically control circulation of uncharacterised environmental strains in commercial poultry flocks.     

Detection of Campylobacter jejuni strains in the water lines of a commercial broiler house and their relationship to the strains that colonized the chickens

Campylobacter jejuni is frequently present in the intestinal tract of commercial broiler chickens, and their drinking water has been proposed to be an initial source of bacteria for newly hatched chicks. We studied three sequential commercial broiler flocks raised in a house from which we had cultured C. jejuni from the nipple waters prior to placement of the first flock. Campylobacter cells were detected by immunofluorescence in the biofilm of the drinking nipples during the weeks when the flock was colonized with C. jejuni but not during weeks when the birds were negative. Campylobacter jejuni was isolated from the drinking water during the growth of the first flock and was present in the birds from all three flocks. Randomly amplified polymorphic DNA (RAPD)-polymerase chain reaction (PCR) typing with primer OPA11 indicated that seven distinct strains were present within the broiler house. One strain found in drinking water was similar to a strain found in birds in the second flock; however, RAPD-PCR with primer HLW85 showed that the strains were not identical. These results suggest that although the watering system is a potential source of C. jejuni in broiler flocks, the waterborne strain in this study was not detected in the birds.