Campylobacter jejuni infection in the ferret: an animal model of human campylobacteriosis

Campylobacter infection in weanling ferrets (Mustela putorius furo) was studied as an animal model for enteric campylobacteriosis in persons. The screening of fecal cultures on selective campylobacter media showed that Campylobacter jejuni/coli was not present in the normal enteric flora. Intragastric feeding of a mixture of cat feed and 2.5 X 10(8) C jejuni isolated from ferrets with naturally occurring proliferative colitis was accomplished. All ferrets (n = 8) became infected on 3 days after they were inoculated, and at 5 to 7 days, they had bile-tinged, liquid feces with excessive mucus and blood. Ferrets gradually recovered from the diarrhea, and feces were normal 10 to 14 days after inoculation was done. Feces contained C jejuni at 14, 23, 28, 39, 46, 60, 91, 101, 109 and 144 days. In the second experiment, weanling ferrets initially were treated with 10% sodium bicarbonate, and 1 X 10(10) C jejuni organisms were administered in the cat feed. Diarrhea with fecal leukocytes and occult blood with occasional mucus appeared in almost all of the 21 ferrets from days 4 through 7. Campylobacter jejuni was isolated from the blood of 11 ferrets between 3 hours and 14 days after they were inoculated. Campylobacter jejuni bactericidal antibodies were present in serum samples at 14 days, with titers of 1:16 to 1:32. Intestinal lesions including cellular infiltration with mononuclear and polymorphonuclear leukocytes were in the lamina propria of the pyloric mucosa and small intestine of infected and control ferrets. The colon of 3 infected ferrets had small focal infiltrates of neutrophils on the lamina propria; one ferret had perivascular cuffing.(ABSTRACT TRUNCATED AT 250 WORDS)     

Reproductive failure in mink and ferrets after intravenous or oral inoculation of Campylobacter jejuni.

Four pregnant mink and seven pregnant ferrets, including five with previous exposure and specific antibody, were injected intravenously with 10(8)-10(10) colony-forming units of Campylobacter jejuni. All 11 pregnancies failed 1-16 days after infection, with results ranging from fetal resorption to expulsion of dead or premature living kits. In every case, uterine contents (placenta, uterine fluid and/or kits) were culture-positive for C. jejuni. Three pregnant mink and nine pregnant ferrets, including four with previous exposure and antibody, were fed 10(9)-10(11) C. jejuni. Two of the mink aborted; kits of all three were culture-positive, but those of one female survived. Seven of the nine ferrets aborted, with two having culture-positive uterine contents. None of 28 uninfected ferret control pregnancies ended in abortion. The most prominent histological feature observed was severe placentitis, which appears to be a more likely cause of Campylobacter-induced abortion than direct pathogenic effects on infected kits. These results suggest that infection of mink or ferrets with C. jejuni during pregnancy poses a serious risk of reproductive failure, even for previously exposed females.     

Campylobacter in the dog: a clinical and experimental study

Faecal samples from 54 dogs with diarrhoea and 54 control dogs were cultured for Campylobacter, Salmonella and Yersinia species and controlled for enteric viruses. The campylobacter were identified as either C jejuni/coli or C upsaliensis. In the diarrhoeic group 16 dogs (29.6 per cent) were positive for campylobacter, 10 C upsaliensis and six C jejuni/coli. Concomitant infection with parvovirus was evident in six of the dogs with diarrhoea and campylobacter-positive faecal cultures. In the control group 13 dogs (24.1 per cent) were positive for campylobacter; three of the isolates were C upsaliensis and six C jejuni/coli. Four isolates could not be identified. The most prominent clinical findings in naturally occurring cases were an acute onset of vomiting (12 of 16), diarrhoea (16 of 16) which was often haemorrhagic (nine of 16) and a raised rectal temperature. Dogs were infected experimentally with both C jejuni (three dogs) and C upsaliensis (three dogs). The challenge strains could be identified in faecal samples from all the dogs, but clinical signs of diarrhoea were seen in only one dog infected with C jejuni. Soft faeces was passed by one dog infected with C upsaliensis. It is concluded that C jejuni/coli or C upsaliensis are either primary pathogens or, after predisposing factors such as virus infections, act as secondary pathogens. It also seems probable that Campylobacter species are present in the intestinal flora of the normal dog.     

Dose response and organ invasion of day-of-hatch Leghorn chicks by different isolates of Campylobacter jejuni

Colonization of the ceca and organ invasion by different isolates of Campylobacter jejuni were investigated in day-of-hatch leghorn chicks. This model of Campylobacter colonization of the ceca demonstrates that 1) day-of-hatch birds do not naturally contain cecal Campylobacter, 2) ceca can be colonized with C. jejuni by oral gavage and not by cloacal inoculation; 3) C. jejuni can be recovered from the ceca up until at least 7 days postinoculation, 4) cecal colonization occurs when as little as 10(2) colony-forming units is orally inoculated into chicks, and 5) different C. jejuni isolates vary both in their ability to colonize the ceca and in their ability to invade the liver. These studies demonstrate that we have a working animal model for Campylobacter colonization for day-of-hatch chicks. This animal model is being used to examine intervention strategies such as vaccines by which Campylobacter can be reduced or removed from the food animal.     

Campylobacter jejuni is associated with, but not sufficient to cause vibrionic hepatitis in chickens

Vibrionic hepatitis is a disease of poultry which is characterised by the presence of focal lesions in the liver, usually 1-2mm in size and greyish-white in colour. The cause of the disease remains unclear, as do the reasons for its recent re-emergence. We examined the livers of commercial broiler chickens taken during processing and found Campylobacter spp. in both normal livers and those displaying signs indicative of focal hepatitis. Livers with signs of hepatitis had significantly more Campylobacter spp. present than those without and other bacterial genera were infrequently present. We were unable to replicate the disease in a healthy host following experimental infection with a Campylobacter jejuni strain isolated from a liver showing signs of focal hepatitis. However, a significant T cell response to C. jejuni was seen in the liver of Campylobacter infected birds. We conclude that the presence of Campylobacter spp. in the liver alone is not sufficient to cause vibrionic hepatitis, but that a predisposing factor, possibly within the host is required. We also provide evidence that chickens mount an adaptive T cell response to systemic C. jejuni.     

Serum agglutinin titers against somatic and flagellar antigens of Campylobacter jejuni and isolation of Campylobacter spp in chickens

From June 1983 to June 1984, two hundred twenty-five 3- to 30-month-old chickens (hens) on 10 different farms were examined for Campylobacter spp. Watering trays and fly vectors also were examined for Campylobacter spp on 6 of the 10 farms. Campylobacter jejuni was isolated from fecal specimens from 64 hens (28.4%), C coli was isolated from 6 hens (2.7%), and C laridis was isolated from 9 hens (4%). The isolation rate of C jejuni was 6.7% to 46.7% for 9 of the 10 farms. On 2 farms, agglutinin titers greater than or equal to 1:40 against somatic and flagellar antigen of C jejuni were detected in hens from which the bacteria were isolated. Hens having titers greater than or equal to 1:40 against C jejuni or hens from which C jejuni had been isolated often occupied adjacent pens. Campylobacter jejuni was isolated from a watering tray on 1 farm and from fly vectors on 2 farms.     

Fla-DGGE analysis of Campylobacter jejuni and Campylobacter coli in cecal samples of broilers without cultivation

In a commercial broiler flock during rearing multiple genotypes of Campylobacter jejuni may be present as well as in gastrointestinal tracts of individual birds. The aim of this study was to optimize and apply a denaturing gradient gel electrophoresis assay of the flagellin gene (fla-DGGE) for analysis of C. jejuni and Campylobacter coli in cecal samples of broilers without prior cultivation. One C. coli and 21 C. jejuni strains isolated from broiler flocks, of which 14 typed as unique by restriction fragment length polymorphism of flaA and two undefined strains, were clustered into 9 groups when applying fla-DGGE. Spiking of cecal samples revealed that fla-DGGE is able to detect at least 4.55-5.96logCFUCampylobacter/mlcecal material. The presence of 3 strains spiked in cecal material was demonstrated by fla-DGGE as the corresponding bands were visible on the DGGE gel. Naturally contaminated cecal samples were shown to contain different types of C. jejuni and C. coli. Fla-DGGE has some potential as a cultivation-independent fast primary subtyping method for C. jejuni and C. coli in cecal samples of broilers.     

Campylobacter jejuni, Campylobacter coli, and cytolethal distending toxin genes in laying hens

As no data are available on the prevalence of cytolethal distending toxin (cdt) genes carried by Campylobacter spp. in laying hens, this study was conducted with the aim to evaluate the prevalence of both Campylobacter spp. and cdt genes in 1680 laying hens from four different farms. The samples were analyzed by culture methods and by polymerase chain reaction. Campylobacter spp. were isolated from 1097/1680 cloacal swabs. Among the isolates, 913 were identified as Campylobacter jejuni whereas 345 were identified as Campylobacter coli. All isolates carried cdt genes. The results presented here confirm the very common occurrence of C. jejuni and C. coli in laying hens and underline that the cdt genes may also be frequently present in both C. jejuni and C. coli isolates from laying hens.     

The occurrence and characterization of Campylobacter jejuni and C. coli in organic pigs and their outdoor environment

The occurrence and species distribution of thermophilic Campylobacter was investigated in organic outdoor pigs. An increased exposure of outdoor pigs to C. jejuni from the environment may cause a shift from a normal dominance of C. coli to more C. jejuni, which may imply a concern of reduced food safety. Bacteriological methods for determination of Campylobacter excretion level were combined with colony-blot hybridization and real-time PCR for specific detection of C. jejuni in pigs. Campylobacter was isolated from pigs (n=47), paddock environment (n=126) and wildlife (n=44), identified to species by real-time PCR and sub-typed by serotyping (Penner) and pulse-field gel electrophoresis (PFGE) genotyping. All pigs excreted Campylobacter (10(3)-10(7) CFU g(-1) faeces) from the age of 8-13-weeks old. C. jejuni was found in 29% of pigs in three consecutive trials and always in minority to C. coli (0.3-46%). C. jejuni and C. coli were isolated from 10% and 29% of the environmental samples, respectively, while crow-birds and rats harboured C. jejuni. Individual pigs hosted several strains (up to nine serotypes). The paddock environment was contaminated with C. coli serotypes similar to pig isolates, while most of the C. jejuni serotypes differed. C. jejuni isolates of different origin comprised few similar serotypes, just one identical genotype was common between pigs, environment and birds. In conclusion, the occurrence of C. jejuni varied considerably between the three groups of outdoor pigs. Furthermore, transfer of C. jejuni to the outdoor pigs from the nearby environment was not predominant according to the subtype dissimilarities of the obtained isolates.     

Enteric colonisation following natural exposure to Campylobacter in pigs

A survey was conducted to establish the prevalence of Campylobacter in pigs from an integrated commercial hog farm. This study was carried out in four different groups of pigs: 1) adult gilts (50); 2) pregnant sows (9); 3) piglets at day-of-birth (73); 4) weaned piglets (20). Rectal and/or caecal samples were collected from each pig. Campylobacter was cultured and enumerated from such samples using Bolton enrichment broth and Campy-Cephex agar plates. Both biochemical and serological tests were used to determine Campylobacter species. Gilts had a 76 per cent incidence of Campylobacter with a mean of 76.3 per cent for C. jejuni, 21 per cent for C. coli and 2.6 per cent for C. lari. Pregnant sows had a 100 per cent incidence of Campylobacter with a mean of 87 per cent for C. jejuni and 13 per cent for C. coli. Newborn piglets had a 57. 8 per cent incidence of Campylobacter, rising to 100 per cent by the time of weaning. Thus it appears that pigs, from the day of birth, are highly susceptible to colonisation by Campylobacter.     

Presence of Campylobacter jejuni in various organs one hour, one day, and one week following oral or intracloacal inoculations of broiler chicks

Day-old broiler chicks (n=30) were obtained from a commercial hatchery and inoculated, either orally or intracloacally, with a characterized strain of Campylobacter jejuni. At 1 hr, 1 day, and 1 wk after inoculation, broilers (n = 5) from the orally and intracloacally inoculated groups along with control birds (n=4) were humanely killed by cervical dislocation. The broilers from the control and treatment groups were aseptically opened, and the thymus, spleen, liver/gallbladder, bursa of Fabricius, and ceca were aseptically removed and individually analyzed for C. jejuni. Overall, C. jejuni was isolated after oral inoculation from 13% (10/ 75), 17% (13/75), and 28% (14/50) of the 1-hr, 1-day, and 1-wk samples, respectively. Campylobacter jejuni was isolated from 10% (4/ 40), 8% (3/40), 10% (4/40), 25% (10/40), and 40% (16/40) of the thymus, spleen, liver/gallbladder, bursa of Fabricius, and ceca samples, respectively. Following the intracloacal route of inoculation, C. jejuni was recovered from 32% (24/75), 8% (6/75), and 16% (8/50) of the 1-hr, 1-day, and 1-wk samples, respectively. Campylobacter jejuni was isolated from 5% (2/40), 5% (2/40), 5% (2/40), 45% (18/40), and 40% (16/40) of the thymus, spleen, liver/gallbladder, bursa of Fabricius, and ceca samples, respectively, for all sampling periods. Campylobacter spp. were not recovered from sample sites examined from the control broilers from trial one, trial two, or trial three samples examined after 1 hr and 1 day. However, one control sample was positive from the 1-wk sampling from repetition three; therefore, those data were omitted. The rapid movement of Campylobacter to internal organs following both oral and intracloacal inoculation may be significant, particularly if it persists in these organs as reservoirs throughout the 65-wk life cycle of breeding birds.     

Dissemination of antimicrobial resistant strains of Campylobacter coli and Campylobacter jejuni among cattle in Washington State and California

The purpose of this study was to investigate the genetic similarity of Campylobacter jejuni and Campylobacter coli with similar antimicrobial resistance phenotypes, isolated from cattle on different farms and at different times, in order to evaluate the possible existence of disseminated antimicrobial resistant clones. PFGE after SmaI and KpnI restriction identified 23 and 16 distinct PFGE patterns among 29 C. jejuni and 66 C. coli isolates, respectively. In C. coli, 51 (77%) of the resistant isolates demonstrated one of the four indistinguishable PFGE patterns, whereas only 24% doxycycline resistant C. jejuni shared one of the two indistinguishable PFGE patterns. The genetic mechanisms of resistance were homogeneous within and between these clonal types. Genetically indistinguishable (clonal) groups of C. coli accounted for most Campylobacter sp. with multiple antimicrobial resistance observed in this study, consistent with a role for clonal dissemination in the epidemiology of resistance in this species.     

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.     

Antimicrobial resistance in Campylobacter spp. isolated from Ontario sheep flocks and associations between antimicrobial use and antimicrobial resistance

The objectives of this study were to determine the prevalence of antimicrobial resistance (AMR) in faecal Campylobacter spp. from lambs and adult sheep and associations between antimicrobial use (AMU) and AMR. A total of 275 faecal samples collected during initial and final visits from 51 sheep flocks, including one feedlot, across southern Ontario were tested for the presence of Campylobacter spp. Campylobacter jejuni was detected in 52% (143/275) of the faecal samples, Campylobacter coli in 7% (19/275), Campylobacter lari in 1% (2/275) and 2% (4/275) were non-speciated Campylobacter. Broth microdilution was used to test antimicrobial susceptibility of 162 isolates to nine antimicrobials. Campylobacter jejuni isolates (n = 142) were resistant to tetracycline (39%), ciprofloxacin (4%), nalidixic acid (4%) and telithromycin (1%). C. coli isolates (n = 19) were resistant to tetracycline (74%), and azithromycin, clindamycin, erythromycin, and telithromycin (5%). The C. lari isolate displayed resistance to nalidixic acid. No statistically significant associations were found between AMU and AMR during multivariate modelling in this study.     

Possible relationship of proliferative enteritis in pigs and hamsters

Three- to six-week-old hamsters were orally inoculated with broths containing one of the following cultures: Campylobacter mucosalis; C. hyointestinalis; C. coli; C. jejuni, all of porcine proliferative enteritis origin, or else C. jejuni of hamster origin. Hamsters given the last of those organisms were shown to have colonisation of their intestines by C. jejuni and 36 of 40 developed an acute enteritis. Mild hyperplasia of enterocytes in ileal crypts was evident in one hamster 2 days after it was given C. coli. No other lesions were detected. Further 3-week-old hamsters were orally inoculated with homogenised intestinal mucosa collected from 4 pigs (A-D) affected by proliferative enteritis. Lesions of proliferative enteritis were detected in 7 of 41 hamsters necropsied 10-21 days after being dosed with mucosas B or D. Marked hyperplasia of ileal enterocytes, associated with numerous intracellular Campylobacter-like organisms, were invariably detected in experimentally affected hamsters. No particular Campylobacter sp. was consistently isolated. None of the controls had demonstrable lesions. The results suggested that cross-species transmission of proliferative enteritis was possible from pigs to hamsters. Therefore a common initiating or aetiological agent may be present. No specific organism was identified as filling this role by inoculation of hamsters with pure cultures.     

Immunohistochemical identification of Campylobacter fetus in natural cases of bovine and ovine abortions

An immunohistochemistry (IHC) procedure for the detection of Campylobacter fetus antigens using an avidin-biotin complex technique was performed on formalin fixed bovine and ovine fetal tissues from 26 natural cases of Campylobacter spp. abortion (four ovine and 22 bovine). The species of Campylobacter isolated included C. fetus ssp. venerealis from 13 bovine fetuses, C. fetus ssp. fetus from two ovine and one bovine fetus, Campylobacter jejuni from seven bovine fetuses, Campylobacter lari from two ovine fetuses and an unspeciated Campylobacter species in one bovine fetus. Histologic lesions identified in the aborted fetuses included placentitis, serositis, pneumonia, gastroenteritis, hepatitis and encephalitis. Campylobacter fetus antigens were identified by IHC in 13 of 13 bovine fetuses from which C. fetus ssp. venerealis was isolated and in two of two ovine fetuses from which C. fetus ssp. fetus was isolated. The IHC stains were negative in tissues from seven bovine fetuses from which C. jejuni was isolated, one bovine fetus infected with C. fetus ssp. fetus, one bovine fetus infected with the unspeciated Campylobacter and two ovine fetuses infected with C. lari. In positive cases, the IHC stain most frequently identified bacteria in the lung and gastrointestinal tract. The C. fetus IHC procedure performed on formalin fixed tissues is a practical tool for the diagnosis of natural cases of ovine and bovine abortion caused by C. fetus.     

Isolation of Campylobacter jejuni and Campylobacter coli from the gall bladder samples of sheep and identification by polymerase chain reaction

In this study, 100 gall bladder samples of sheep slaughtered at an abattoir in Elazi? province were examined for Campylobacter jejuni and Campylobacter coli by culture and polymerase chain reaction (PCR). Preston Campylobacter Agar supplemented with 7% horse blood and Preston Selective Supplement (Oxoid, Hampshire, UK) were used for isolation of the agents. Campylobacter spp. were isolated in 66 samples, and they were identified as 34% C. jejuni and 32% C. coli. A multiplex PCR based upon the use of ceuE gene-specific primers was applied on DNA samples extracted from C. jejuni and C. coli isolates. All C. jejuni and C. coli strains that were positive by culture were also detected to be positive by PCR. This study shows that PCR can be used an alternative, rapid and sensitive test for the identification of C. jejuni and C. coli which threaten human and animal health.     

Experimental infection of specific pathogen-free pigs with Campylobacter: excretion in faeces and transmission to non-inoculated pigs

Campylobacter species are leading agents of human bacterial gastroenteritis and consumption of food of animal origin is a major source of infection. Although pigs are known to frequently exhibit high counts of Campylobacter in their faeces, more information is needed about the dynamics of this excretion. An experimental trial was conducted to evaluate the faecal excretion of Campylobacter by 7-week-old specific pathogen-free piglets inoculated per os with three Campylobacter strains (one C. coli isolated from a pig, one C. coli and one C. jejuni from chickens) alone or simultaneously (5x10(7)CFU/strain). Non-inoculated pigs were housed in adjacent pens. Pigs were monitored for 80 days for clinical signs and by bacteriological analysis of faeces. Pigs inoculated with porcine C. coli or with a mix of the three strains excreted from 10(3) to 10(6)CFU/g of faeces with a slight decrease at the end of the trial. Animals inoculated with poultry C. coli or C. jejuni strain excreted a lower quantity and some of them stopped excreting. At the end of the trial, only C. coli was detected in the faeces of pigs inoculated simultaneously with the three bacteria. Moreover, the transmission of Campylobacter was noticed between pens for the two C. coli strains and all the neighbouring animals became shedders with a level of excretion similar to the inoculated pigs. Intermittence in the Campylobacter excretion was also observed. Finally, our study highlighted a host preference of Campylobacter, namely C. coli seems to have a higher colonization potential for pigs than C. jejuni.     

The occurrence and characterization of Campylobacter spp. in silver gulls (Larus argentatus), three-toed gulls (Rissa tridactyla) and house sparrows (Passer domesticus)

Altogether 16 Campylobacter (C.) isolates could be recovered from 65 Herring gulls: 5 x C. laridis, 2 x C. jejuni biovar 1, 4 x C. jejuni biovar 2 and 5 x C. coli. Campylobacter spp. were isolated from 15 out of 51 samples from Kittiwakes: 2 x C. jejuni biovar 1 and 13 x C. laridis. All C. coli isolates grew on agar containing 1.5% NaCl. Two Campylobacter isolates from 50 House sparrows differed from all other isolates by a distinct beta-hemolysis and other phenotypic characteristics and could not be associated with a certain Campylobacter species. Epidemiological aspects and the possible role of the examined birds as a source of infection for man and domestic animals are discussed.     

Campylobacter spp. in dogs and cats in Switzerland: risk factor analysis and molecular characterization with AFLP

A 1-year cross-sectional study was carried out to determine the prevalence, risk factors for carriage, and genetic diversity of Campylobacter spp. in healthy dogs and cats in Switzerland. Veterinary practitioners collected samples from 1268 animals (all ages) presented for vaccination. The prevalence of Campylobacter spp. in 634 dogs and 596 cats that were eligible for the study was 41.2% (confidence interval 95%: 37.3-45.1%) and 41.9% (CI 95%: 37.9-46%), respectively. Risk factors identified for carriage of Campylobacter jejuni were found to be different from risk factors for C. upsaliensis/C. helveticus. Young animals (< or =3 years) had significantly higher odds of carrying C. upsaliensis/C. helveticus than older animals (OR 1.8-3.3), whereas for C. jejuni carriage, the age was not a risk factor. Amplified fragment length polymorphism (AFLP) genotyping revealed heterogeneity among C. jejuni strains and was found to clearly separate C. helveticus from C. upsaliensis. It was shown that cats more often carry C. helveticus with an estimated prevalence of 28.2%, whereas dogs mainly are carrying C. upsaliensis.