Genotype dynamics of Campylobacter jejuni in a broiler flock

We investigated the genotype diversity and dynamics of Campylobacter in a commercial broiler flock during rearing and slaughter. In total, 220 Campylobacter jejuni isolates collected on four sampling occasions during rearing and from routine sampling during slaughter were subtyped by SmaI macrorestriction and pulsed-field gel electrophoresis, PFGE. Eight different SmaI types were found. During rearing, a subsequent addition of genotypes occurred, with two SmaI types found at 2 weeks of age and six types on the day before slaughter. All types that were detected in more than one isolate were also found on all succeeding sampling occasions, including the slaughter sampling. Two new types were found in the slaughter samples. In two-thirds of the individual birds sampled the day before slaughter, more than one SmaI type were found, although there was a clear tendency for dominance of one type in individual birds. Our results show that multiple genotypes of C. jejuni may be present in a commercial broiler flock during rearing and even in gastrointestinal tracts of individual birds. Both recurring environmental exposure and genetic changes within the population may explain the genotype diversity. Although the distribution of genotypes varied between different sampling occasions, we found no indication that any subtype excluded another during the rearing of the broiler flock.     

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.     

Dynamics of populations of Campylobacter jejuni in two grandparent broiler breeder farms: persistent vs. transient strains

The objectives of the study were to characterize and investigate the populations of Campylobacter jejuni in two grandparent broiler breeder farms over four years. Caecal as well as farm environmental samples were obtained. Campylobacter isolates were characterized by macrorestriction profile (SmaI and KpnI-PFGE) and PCR-RFLP of the flaA gene. Susceptibility tests against seven antimicrobials were also performed. Birds were negative for Campylobacter spp. when they came to these two production farms (20 weeks), and most of the flocks remained uncolonized until they were 23 weeks old. Eighteen genotypes were characterized, with one of them (genotype 2) appearing and persisting over the study period in the two farms. In general, the strains exhibited high genetic stability, and most of them could be seen as transient in the farms, being substituted by other strains when their flock was substituted. Only one environmental sampling was positive for C. jejuni. Two different genotypes were characterized; one of them was isolated from the birds of that farm two years before. The susceptibility data point to the idea of an environmental source or reservoir of this genotype. Regarding the susceptibility of the populations, as other studies have shown, quinolone resistance (alone or combined with other resistances) was the most frequent: 68.5%. Quinolone- and multidrug-resistant strains are a matter of concern in public health. In conclusion, this survey shows the complexity of the study of the colonization of farms by C. jejuni.     

Genetic comparison of Campylobacter jejuni isolated from different cattle farms

To compare the genotypes of Campylobacter jejuni, isolates of cattle origin were collected from 9 Polish farms and genotyped by ERIC-PCR. We identified 28 genotypes among the 43 C. jejuni isolates, and demonstrated high genomic diversity. The highest level of diversity was observed in strains isolated from stanchion-barn animals in opposition to those from the loose-housing system.     

Risk factors for Campylobacter jejuni and Campylobacter coli in young cattle on English and Welsh farms

Campylobacter jejuni and C. coli are the most prevalent causes of bacterial diarrhoea in most of the Western World. In Great Britain, the source remains unknown for the majority of cases, though poultry is considered the main source of infection. Molecular typing methods identify cattle as a potential source of a proportion of the non-source-attributed cases, mainly through direct contact, environmental contamination or milk, but little is known about the epidemiology of Campylobacter in cattle. A cross-sectional study was undertaken on young cattle 3-17 months of age on 56 cattle farms in England and Wales to identify association between the presence of C. jejuni and C. coli and farm characteristics and management practices. Campylobacter was detected on 62.5% of the farms and the presence of dairy cows (OR: 3.7, CI(95%): 1.2; 11.7), indoor housing (OR: 4.6, CI(95%): 1.8; 12.0), private water supply (OR: 2.5, CI(95%): 1.2; 5.4), presence of horses (OR: 3.2, CI(95%):1.5; 6.9) and feeding hay (OR: 2.9, CI(95%):1.6; 5.5) were associated with detection. The model's goodness-of-fit was improved when herd size was forced in the model without being statistically significant (p=0.34).     

Passive immunization to reduce Campylobacter jejuni colonization and transmission in broiler chickens

Campylobacter jejuni is the most common cause of bacterium-mediated diarrheal disease in humans worldwide. Poultry products are considered the most important source of C. jejuni infections in humans but to date no effective strategy exists to eradicate this zoonotic pathogen from poultry production. Here, the potential use of passive immunization to reduce Campylobacter colonization in broiler chicks was examined. For this purpose, laying hens were immunized with either a whole-cell lysate or the hydrophobic protein fraction of C. jejuni and their eggs were collected. In vitro tests validated the induction of specific ImmunoglobulinY (IgY) against C. jejuni in the immunized hens’ egg yolks, in particular. In seeder experiments, preventive administration of hyperimmune egg yolk significantly (P < 0.01) reduced bacterial counts of seeder animals three days after oral inoculation with approximately 10⁴ cfu C. jejuni, compared with control birds. Moreover, transmission to non-seeder birds was dramatically reduced (hydrophobic protein fraction) or even completely prevented (whole-cell lysate). Purified IgY promoted bacterial binding to chicken intestinal mucus, suggesting enhanced mucosal clearance in vivo. Western blot analysis in combination with mass spectrometry after two-dimensional gel-electrophoresis revealed immunodominant antigens of C. jejuni that are involved in a variety of cell functions, including chemotaxis and adhesion. Some of these (AtpA, EF-Tu, GroEL and CtpA) are highly conserved proteins and could be promising targets for the development of subunit vaccines.     

Prevalence and clonal diversity of Campylobacter jejuni from dairy farms and urban sources

AIM: To investigate the role of free-living animals such as sparrows, rodents and flies as potential reservoirs of Campylobacter spp on a dairy farm, and to assess the genetic diversity among Campylobacter isolates from the farm and an urban source. METHODS: A total of 290 samples (bovine, passerine and rodent faeces, and whole flies) were collected from a large commercial dairy farm in the Manawatu district in New Zealand, and from faeces from urban sparrows in a nearby city. Other samples collected from the dairy farm included five from silage, two from aprons worn by workers during milking, two from workers' boots and two from water in troughs in a paddock. Isolates of thermophilic Campylobacter spp were identified morphologically and phenotypically and further characterised molecularly using pulsed-field gel electrophoresis (PFGE) and the restriction enzyme SmaI. RESULTS: Campylobacter jejuni was the only Campylobacter species isolated from all samples. The highest prevalence was found in faeces from dairy cows (54%), followed by faeces from sparrows from the urban area (40%) and the farm (38%), and from rodents (11%) and whole flies (9%). Other samples from the farm environment such as silage, trough water, and workers' aprons and boots were also positive for C. jejuni. Of the 22 restriction patterns obtained, seven were common to more than one source. CONCLUSIONS: Cattle, sparrows, rodents and flies are potential reservoirs of C. jejuni on dairy farms. Identical clones of C. jejuni carried by cattle, sparrows, flies and rodents probably indicate a common source of infection. The high level of asymptomatic carriage of C. jejuni by healthy dairy cows could be sufficient to maintain infections within the dairy farm surroundings via environmental contamination.     

Prevalence and clonal diversity of Campylobacter jejuni from dairy farms and urban sources

AIM: To investigate the role of free-living animals such as spar- rows, rodents and flies as potential reservoirs of Campylobacter spp on a dairy farm, and to assess the genetic diversity among Campylobacter isolates from the farm and an urban source.

METHODS: A total of 290 samples (bovine, passerine and ro- dent faeces, and whole flies) were collected from a large com- mercial dairy farm in the Manawatu district in New Zealand, and from faeces from urban sparrows in a nearby city. Other samples collected from the dairy farm included five from silage, two from aprons worn by workers during milking, two from workers' boots and two from water in troughs in a paddock. Isolates of thermophilic Campylobacter spp were identified mor- phologically and phenotypically and further characterised mo- lecularly using pulsed-field gel electrophoresis (PFGE) and the restriction enzyme SmaI.

RESULTS: Campylobacter jejuni was the only Campylobacter species isolated from all samples. The highest prevalence was found in faeces from dairy cows (54%), followed by faeces from sparrows from the urban area (40%) and the farm (38%), and from rodents (11%) and whole flies (9%). Other samples from the farm environment such as silage, trough water, and work- ers' aprons and boots were also positive for C. jejuni. Of the 22 restriction patterns obtained, seven were common to more than one source.

CONCLUSIONS: Cattle, sparrows, rodents and flies are po- tential reservoirs of C. jejuni on dairy farms. Identical clones of C. jejuni carried by cattle, sparrows, flies and rodents prob- ably indicate a common source of infection. The high level of asymptomatic carriage of C. jejuni by healthy dairy cows could be sufficient to maintain infections within the dairy farm sur- roundings via environmental contamination.     

Effect of coated and non-coated fatty acid supplementation on broiler chickens experimentally infected with Campylobacter jejuni

The aim of this study was to examine whether and to what extent the supplementation of feed with a coated or non-coated mixture of fatty acids (caprylic and capric acid) affects broiler chickens experimentally infected with Campylobacter jejuni. The study was carried out using 48 chickens divided into four experimental groups. Throughout the whole rearing period (1-42 days), the chickens were fed a diet supplemented with 0.25% caprylic and capric acid (1:1), coated or non-coated. At the age of 14 and 28 days, chickens were orally challenged with C. jejuni. At regular time intervals post-inoculation, the shedding of C. jejuni was assayed using quantitative real-time PCR. Both supplements significantly decreased faecal C. jejuni counts by 1.2-4.1 log(10) CFU/g 4 days post-inoculation; after this time period, the effect of medium-chain fatty acids (MCFA) was less pronounced or absent. Campylobacter jejuni counts in excreta samples were significantly lower in chickens fed coated MCFA than in those fed non-coated MCFA. No effect of MCFA on feed intake or growth of chickens was observed. In conclusion, (i) MCFA are active against C. jejuni and (ii) the encapsulation enhanced the efficacy of the acids. These results allow the recommendation of using MCFA as feed additives in chickens, preferably 2-3 days before slaughter.     

The viability of Campylobacter jejuni on refrigerated chicken drumsticks

Radappertized chicken drumsticks were experimentally contaminated with suspensions of Campylobacter jejuni in two trials. Qualitative analysis on drumsticks with an initial level of contamination of 4.8 X 10(3) CFU/cm2 showed that viability was retained for at least 10 days of storage at either 9 degrees or -12 degrees C. In a second quantitative trial, the level of contamination declined from 9.9 X 10(2) CFU/cm2 to 4.5 X 10(1) CFU/cm2 after 7 days at -20 degrees C. Thereafter, C. jejuni persisted at levels ranging from 1.8 X 10(1) to 0.2 X 10(1) CFU/cm2 through the 26th week of storage. Drumsticks held at 4 degrees C showed a significant decline in count from 9.9 X 10(2) CFU/cm2 to 1.8 X 10(2) CFU/cm2 on day 7. It is concluded that the viability of C. jejuni on chicken parts is maintained under both refrigerated and freezing conditions which approximate commercial storage. This is of significance to the meat industry and consumers.     

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.     

空肠弯曲菌毒力因子研究进展

空肠弯曲菌(Campylobacter jejuni)是世界范围内广泛流行的人畜共患病,在公共卫生学上具有重要意义。空肠弯曲菌可引发人和多种动物疾病,其抗原结构复杂,致病机理尚不清楚。作者通过对空肠弯曲菌可能的毒力因子及其在致病中的作用作一简要概述,为空肠弯曲菌的诊断防治提供依据。     

鸡空肠弯曲菌对SPF雏鸡致病性研究

用鸡空肠弯曲菌（Ｃａｍｐｙｔｏｂａｃｅｃｒｉｅｊｕｎ：）对１日龄ＳＰＦ（Ｓｐｃｉｉｃ－ｐａｔｈｏｇｅｎｆｒｃｅ）雏鸡进行感染实验。随机将６０只１日龄ＳＰＦ雏鸡分为４组，那和对照组，每组１５只。分别用含菌７．５Ｘ１０８ＣＦＵ／ｍｌ、７．５ｘ１０４ＣＦＵ／ｍｌ，７．５×１０２ＣＦＵ／ｍｌ布氏肉汤菌液经口时ｔ、Ｊ、Ｉ组接种。接种后分别于２、４、６，８、１０日踏踏机取３只／组、经剖构观察病理变化；从心血、肝、胆汁、脾、肠内容物分离病原菌，测量肠道内物空肠弯曲菌菌数。得出以下结论；经口接种３．７５ｘ１０２ＣＦＵ空场弯曲菌即可造成１日龄ＳＰＦ雏鸡感染发病；感染鸡的主要病理变化为肝脏表面有点状，条索状或片状出血，同时伴有形状不规则大小不等的黄白色坏死灶，盲肠膨大充满气泡，十二指肠、空肠、直肠粘膜有散在的小出血点，首选检苗材料为胆汁、其次是脾、肝等器宫。     

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.     

Effects of diet formulations containing proteins from different sources on intestinal colonization by Campylobacter jejuni in broiler chickens

The objective of this study was to compare the effects of 3 diet formulations containing different protein sources (animal, plant, and a combination of animal and plant) on the colonization of Campylobacter jejuni in the gastrointestinal tract of broiler chickens. A freshly isolated strain of C. jejuni (biotype IV, serotype HS O:21, O:29, HL untypable) from a broiler chicken was used to infect 3-day-old chicks that had been free of C. jejuni; 0.5 mL of an inoculum containing 10⁸ colony-forming units was administered orally. Shedding of the organism was studied, and C. jejuni in the ceca, jejuni, and crop were enumerated by quantitative culture. The isolates recovered from the birds during the study period of 35 d were characterized and confirmed as C. jejuni by the use of standard methods and underwent biotyping, serotyping, antimicrobial susceptibility testing by disk diffusion and the E-test, and flagellin gene typing. A cyclical pattern of shedding of C. jejuni was observed in all the birds. Colonization was highest in the ceca. The ceca of birds receiving plant-protein-based feed had significantly less colonization then the ceca of birds receiving the other types of feed, whereas the differences in colonization of the jejuni and crops were not significant. Characterization by biotyping, serotyping, and flagellin gene typing showed that 95% of the recovered isolates were identical to the strain used for infecting the chicks. However, with the Lior-HL typing scheme, 74% of the recovered isolates were HL untypable. Antimicrobial resistance testing did not reveal significant differences between the infecting strain and the recovered isolates among the different feed groups.