Campylobacter jejuni infections in gnotobiotic pigs

At 3 days of age, 12 gnotobiotic pigs were inoculated orally with broth cultures of Campylobacter jejuni. One pig was euthanatized and evaluated each day for 12 days. In the cecum and colon, there was diffuse edema, neutrophilic infiltration, and sloughing of epithelial cells from the mucosa on postinoculation days (PID) 2 through 5. Dysplastic colonic crypt epithelial cells were observed in the submucosa of the colon on PID 5 through 12. Curved, rod-shaped bacteria were detected on the surface of ileal, cecal, and colic absorptive and glandular epithelial cells. Bacteria also were found around small submucosal vessels on PID 3 and 4 and were associated with numerous perivascular neutrophils. The gnotobiotic pig appears to provide a simple, well-controlled in vivo model for the study of the pathogenesis of C jejuni infections in human beings, pigs, and other mammals.     

Correlation between invasion of Caco-2 eukaryotic cells and colonization ability in the chick gut in Campylobacter jejuni

In an in vitro cell culture model using Caco-2 cells the adhesion and invasion properties of 11 Campylobacter (C.) jejuni isolates of different origin were studied. Additionally, we investigated the colonization ability of the strains in a chick model. Virtually, all C. jejuni showed cell adherence in the in vitro assay, but there were large differences in the invasion frequencies among the Campylobacter isolates. The colonization ability in the chick gut also differed markedly and enabled the formation of three groups: non-colonizing, weak or delayed colonization and strong colonization ability. On this occasion, we found a putative correlation between invasion of Caco-2 cells and colonization in the chick gut. Non-colonizers are not invasive or only have small invasion indexes. Strains which colonize weakly or exhibit delayed colonization have a medium invasion index and strong colonizers show markedly higher values of this parameter. The characterization of the flagellin gene of the used C. jejuni strains resulted in eight flaA types. There was no association between flaA type and invasion or colonization ability in the chick gut.     

PCR detection of virulence-associated genes in Campylobacter jejuni strains with differential ability to invade Caco-2 cells and to colonize the chick gut

In this study, the presence of 20 putative virulence genes was examined in 11 Campylobacter jejuni isolates with different colonization and invasion abilities as determined in a chick colonization model and on Caco-2 cells, respectively. The majority of the genes were detected in all strains. Among them, there were genes of the flagellar secretion apparatus like flhA, flhB, flgB, flgE2, the flagellin genes flaA and flaB, invasion-associated genes like ciaB and iamA, the cytotoxin genes cdtA-C, the adhesion related gene cadF, and some genes involved in the colonization process (docA, docB). The plasmid gene virB11 could not be detected in any strain. Specific differences between the isolates were observed only in genes cgtB and wlaN involved in lipo-oligosaccharide (LOS) biosynthesis. The gene cgtB was only detectable in three of five strains with strong colonization and invasion abilities. Probably, wlaN can overcome the lack of cgtB in the two cgtB- isolates.     

Campylobacter jejuni in poultry giblets

A total of 200 poultry giblets, 50 each of chickens, ducks, squab and turkeys, were examined for the presence of Campylobacter jejuni. In chicken giblets, C. jejuni was isolated from gizzards, hearts, livers and spleens with incidences of 28%, 10%, 40% and 16% respectively while 24%, 6%, 36% and 10% of duck gizzards, hearts, livers and spleens were positive for the organism, respectively. C. jejuni was detected in 6% of squab gizzards, in 10% of squab livers but failed to be detected in squab hearts & spleens. In turkey giblets, 16% of gizzards, 4% of hearts, 30% of livers and 8% of spleens were positive for the organism. C. jejuni was more frequently isolated from liver samples than gizzard, spleen and heart samples, each constituting of 29%, 18.5%, 8.5% and 5%, respectively. High incidence of C. jejuni was recorded among chicken giblets (23.5%), followed by duck giblets (19%), then turkey giblets (14.5%) and finally squab giblets (4%).     

Campylobacter jejuni and Campylobacter coli inoculation of neonatal calves

Three groups of neonatal calves were inoculated orally with pathogenic strains of Campylobacter jejuni or C coli. The calves developed a mild, self-limiting enteritis characterized by thick mucoid feces. Bacteremia and fecal shedding of Campylobacter were sporadic in all inoculated calves. Two groups of calves were killed 1 to 3 weeks after inoculation to study the pathogenesis of infection. Postmortem culture of tissues revealed C jejuni or C coli most frequently in the ileum, cecum, colon, and blood. Clinical or pathologic differences between C jejuni-inoculated and C coli-inoculated calves were minimal.     

Campylobacter jejuni contamination of eggs

Contamination of commercial table eggs with a fecal suspension containing 4.4×10⁶ CFU/gCampylobacter jejuni resulted in shell penetration in 3/70 eggs and recovery of the organism from homogenized egg contents in 1/70 eggs. Viability ofC. jejuni on the shell surface was retained for only 16 hours, attributed to desiccation of the fecal suspension. A field survey of three commercial laying farms and their associated egg-packing plants showed that hens demonstrated to be fecal shedders ofC. jejuni (12% to 62% incidence) did not produce infected eggs. The organism could not be detected in the environment of the packing plant, including grading machinery and effluent.     

Campylobacter jejuni contamination of eggs

Contamination of commercial table eggs with a fecal suspension containing 4.4 X 10(6) CFU/g Campylobacter jejuni resulted in shell penetration in 3/70 eggs and recovery of the organism from homogenized egg contents in 1/70 eggs. Viability of C. jejuni on the shell surface was retained for only 16 hours, attributed to desiccation of the fecal suspension. A field survey of three commercial laying farms and their associated egg-packing plants showed that hens demonstrated to be fecal shedders of C. jejuni (12% to 62% incidence) did not produce infected eggs. The organism could not be detected in the environment of the packing plant, including grading machinery and effluent.     

Campylobacter jejuni infection in broiler chickens

Day-old, straight-run broiler chickens were procured from a hatchery located in the Pacific Northwest. The chickens were subdivided individually into nine groups of 20 chickens. The chickens were tagged, housed in isolation chambers on wire, fed commercial broiler feed, and given water ad libitum. Three isolates of Campylobacter jejuni of poultry origin and one of human origin were tested in this study. Various C. jejuni cultures were inoculated into 9-day-old chickens by crop gavage. Four groups of 20 chickens were inoculated at a dose level of 0.5 ml of 1 x 10(2) colony-forming units (CFU)/ml. The other four groups were inoculated with 0.5 ml of 1 X 10(4) CFU/ml. One group of 20 chickens was kept as an uninoculated control group. Four randomly selected chickens from each of the inoculated and uninoculated groups were necropsied at 5, 12, and 19 days postinoculation (DPI). The C. jejuni was cultured and enumerated from a composite of the upper and midintestine and the cecum. Body weights of all chicken groups at 7 days of age and at 5, 12, and 19 DPI were measured and statistically analyzed. No significant differences were present in the mean body weights (MBWs) of 7-day-old, 5 DPI, and 12 DPI male and female broiler chickens inoculated with C. jejuni at both dose levels compared with uninoculated controls. Differences in MBWs of the male and female broilers at 19 DPI were observed in some of the groups. Results of the C. jejuni culture enumeration mean (CEM) of composite intestine samples at 5 DPI from all inoculated chicken groups, irrespective of the dose level, ranged from (2.5 +/- 5.0) x 10(2) to (2.8 +/- 4.8) x 10(5) CFU/g (mean +/- SD). Results of cecum C. jejuni CEM at 5 DPI inoculated at both dose levels ranged from (2.5 +/- 5.0) x 10(6) to (1 +/- 0.0) x 10(7) CFU/g in all treatment groups irrespective of the dose level. CEM results from the composite intestine samples at 12 and 19 DPI increased by 1 log unit, or sometimes more. Results of cecum C. jejuni CEM at 5 DPI inoculated at both dose levels ranged from (2.5 +/- 5.0) x 10(6) to (1 +/- 0.0) x 10(7) CFU/g in all treatment groups irrespective of the dose level. Increases of 2-5 log units in C. jejuni CEM was present in chicken groups inoculated with 1 X 10(2) CFU of C. jejuni, and a 2- to 3-log increase was present in groups inoculated with a higher dose level of C. jejuni at 12 DPI. The results of C. jejuni CEM from cecal samples at 19 DPI were similar to chicken groups at 12 DPI. Campylobacterjejuni was not isolated from the uninoculated control chickens at 5, 12, and 19 DPI. Clinical signs of illness or gross pathologic lesions were not present in any of the chicken groups during this study. No lesions were present on histopathologic evaluations in C. jejuni-inoculated chickens or uninoculated control chickens.     

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.     

Pathology of Campylobacter jejuni abortion in sheep

Campylobacter jejuni was inoculated intravenously into pregnant ewes on gestation days 114 and 123 to reproduce ovine abortion. All ewes aborted 7-12 days post-inoculation. High numbers of C. jejuni were isolated from ewe tissues (caruncle, bile, cecal feces), fetal tissues, and placenta. C. jejuni colonies were identified in caruncles and placenta by light microscopy and immunoperoxidase techniques. Histologically, inoculated ewes had a severe purulent endometritis with vasculitis. Placentas from inoculated ewes and field cases showed necrosis and purulent inflammation; however, placentas from inoculated ewes had large numbers of bacterial colonies compared to few bacteria found in field cases. Histologically, only one fetus from the inoculated ewes showed lesions (purulent bronchopneumonia), whereas all fetuses from field cases had a distinct bronchopneumonia, and one fetus showed multifocal hepatic necrosis. These results suggest that C. jejuni (serotypes Penner 1 and Lior 2) is an important abortifacient organism for sheep.     

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).     

Campylobacter jejuni in poultry processed in slaughterhouses

The frequency of occurrence of Campylobacter jejuni germs in dressed poultry was studied for a year. The samples--smears from the body cavities of chickens--were collected during the technological dressing of the chickens; 101 strains of Campylobacter jejuni (i. e. 28.69%) were isolated from the 352 samples analyzed. The occurrence of the germs exhibited a considerable seasonal variance with peak rates in spring and summer. The use of a suitable culture medium, the technique of cultivation and the properties of the isolated strains were studied at the same time. The culture medium (Agar no. 3 IMUNA enriched with supplement C, horse blood and ingredients increasing the aerotolerance of the germs--sodium pyruvate and iron sulphate) used during the investigation was found to be suitable. The technique of cultivation by means of an anaerostat manufactured by the Development Station in Brno, atmosphere regulation (5% CO2) and with a pre-set cultivation temperature (43 degrees C) was found to be suitable for the screening of the Campylobacter jejuni germs.     

Heterogeneity of Campylobacter jejuni and Campylobacter coli strains from healthy sheep

The objectives of this study were to identify, at species level, thermophilic campylobacters isolated from clinically healthy sheep by a multiplex polymerase chain reaction (mPCR). The heterogeneity among Campylobacter jejuni and C. coli isolates was also investigated using a restriction fragment length polymorphism (RFLP) analysis of the flagellin (flaA) gene. Samples of intestinal contents, gall bladders and faeces were collected from 610 healthy sheep. While gall bladder samples were plated directly onto Preston agar, an enrichment stage was applied for intestinal and faecal samples. Of the 610 samples, 302 (49.5%) were positive for Campylobacter spp. Using a mPCR assay for species identification, 103 (34.1%) were positive with C. jejuni-specific primers, while 100 (33.1%) were positive with C. coli-specific primers. Additionally, 16 (11.9%) of the intestinal content samples were positive for both species by mPCR. All the isolates identified as C. jejuni and C. coli were successfully subtyped by flaA typing. Of 203 isolates tested, 48 different flaA types were found. Twenty-six flaA types were identified among C. jejuni isolates and the remaining 22 from C. coli isolates.     

Fecal shedding of Campylobacter jejuni and Campylobacter coli among feral pigs in Texas

The population and range of feral pigs in the United States are rapidly expanding, yet key knowledge gaps exist regarding their role in the ecology and transmission of foodborne pathogens. Our objectives were to estimate the prevalence of Campylobacter jejuni and Campylobacter coli shedding among feral pigs throughout Texas and to identify risk factors for positive status. Faecal samples were collected from feral pigs in Texas from February 2014 through May 2015, and target organisms were detected using PCR assays. The prevalence of C. jejuni shedding was 1.6% (6/370), and the prevalence of C. coli shedding was 3.5% (13/370). C. coli shedding was significantly more common (p = .008) among female pigs than among male pigs. Feral pigs may represent a source of human campylobacteriosis.     

猪、鸡间空肠弯曲杆菌感染分布状况的调查研究

目的:对入沪28个产地(河南开封、山东青岛、江苏南通、浙江金华、上海奉贤等地)的鸡、猪进行了空肠弯曲杆菌流行分布调查,了解空肠弯曲杆菌在不同动物种间的分布状况。方法:以空肠弯曲杆菌特异性的VS1基因为扩增目标片段,对2005～2007年间在上海市三家家禽市场和五家生猪屠宰场采集的2234(鸡1015份、猪1219份)盲肠内容物样品进行PCR检测。结果:鸡盲肠内容物检出空肠弯曲杆菌阳性的样品288份,平均阳性率为28.4%,猪盲肠内容物检出空肠弯曲杆菌阳性的样品420份,平均阳性率为34.4%。结论:28个产地的鸡、猪均有空肠弯曲杆菌感染,不同产地、种属其感染率和发病情况不同,经统计学分析显示,猪的感染阳性率显著高于鸡,上海市内(奉贤等地)来源的畜禽感染阳性率显著低于浙江金华、江苏海门、江苏张家港和江苏大丰等外省市地区。