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Development of a new protocol for the isolation and quantification of Arcobacter species from poultry products.

None of the presently available selective supplements for the specific isolation of Arcobacter species allows the growth of Arcobacter butzleri, A. cryaerophilus and A. skirrowii and at the same time fully suppresses the accompanying flora present in poultry and poultry products. Furthermore, little is known about the contamination levels of poultry with Arcobacter species. In this study, a new selective supplement comprising amphotericin B (10 mg/l), cefoperazone (16 mg/l), 5-fluorouracil (100 mg/l), novobiocin (32 mg/l) and trimethoprim (64 mg/l) was developed. With a new isolation procedure, including enrichment in Arcobacter broth with the selective supplement, incubated for 24 to 48 h at 28 degrees C under microaerobic conditions, arcobacters were isolated from 100% (n = 34) of neck skin of laying hens and from 90% (n = 71) of similar samples from broilers. Of the broiler breast meat samples examined (n = 52), 65% were found to be contaminated with these bacteria. In 64% of the samples, A. butzleri was the only Arcobacter species isolated. In 9% of the samples, A. cryaerophilus was the only species present, while 11% of the samples were positive for both species simultaneously. Using direct isolation on the selective agar medium developed in this study, incubated for 24 to 48 h under microaerobic conditions at 28 degrees C. 32 out of 45 broiler carcasses and 6 out of 25 broiler breast meat samples carried a bacterial load of arcobacters of 10(2) to 10(3) cfu/g. The prevalence of Arcobacter in Belgian poultry was found higher than the prevalence of thermophilic Campylobacter species in each of the poultry categories examined. The enrichment procedure and the direct plating method were validated for the isolation of A. skirrowii. For this species, growth performance was less than the other two Arcobacter species and it was not isolated nor detected by m-PCR from the naturally contaminated poultry samples examined. This new protocol provides a fast and reliable method for the isolation of Arcobacter species from poultry and can contribute to more comprehensive epidemiological investigations.

Animals↗

Isolation and characterization of the emerging foodborn pathogen Arcobacter from human stool.

At present, isolation of arcobacters from human specimens is performed by slightly of not modified Campylobacter, Yersinia or Leptospira isolation techniques, and knowledge if arcobacters are part of the human commensal flora is lacking. Therefore, an Arcobacter selective isolation procedure was validated for the examination of human fecal specimens, and the presence and characteristics of Arcobacter in feces of asymptomatic humans was examined in order to assess the clinical relevance of arcobacters in diarrheal stool. With this method, Arcobacter was isolated from seven of 500 (1.4%) stool samples of healthy people with Arcobacter cryaerophilus as the only species present. Seven A. cryaerophilus genotypes were detected and only one genotype was found per person. Neither A. butzleri nor A. skirrowii were isolated, therefore the presence of those latter species in clinical samples requires further attention. Though the pathogenic role and potential virulence factors of arcobacters have to be further examined, the current status of arcobacters as emerging pathogens remains justified.

Adult↗

Occurrence and distribution of Arcobacter species in poultry processing.

A total of 16 broiler flocks slaughtered in the morning in eight Belgian poultry slaughterhouses were examined for the presence of Campylobacteraceae. In samples collected before and after chilling, the prevalence of arcobacters was found to be higher than the prevalence of thermophilic campylobacters, with the slaughter procedure used having no clear effect. Two slaughterhouses were selected for a detailed investigation of the occurrence and distribution of arcobacters. Sampling carried out before slaughter revealed that both Arcobacter butzleri and Arcobacter cryaerophilus were commonly present on the slaughter equipment in both plants. These findings indicate inadequate decontamination of the slaughterhouse environment and suggest potential Arcobacter contamination of broiler carcasses through the slaughter equipment. Even before evisceration, contamination levels of hundreds to several thousands of arcobacters per gram of neck skin were detected. It appears unlikely that contamination through slaughter equipment alone explains the high contamination levels found for poultry products. Arcobacters were not isolated from the 30 intestinal tracts sampled for each broiler flock examined. A. cryaerophilus was the only Arcobacter species recovered from the transport crate samples collected before and after washing. Arcobacter contamination during slaughter, either direct (from chicken intestinal content or feces) or indirect (from equipment), was not confirmed. The origin and the precise routes of contamination remain to be determined.

Abattoirs↗

Genetic diversity of Arcobacter and Campylobacter on broiler carcasses during processing.

Broiler carcasses (n=325) were sampled at three sites along the processing line (prescalding, prechilling, and postchilling) in a commercial poultry processing plant during five plant visits from August to October 2004. Pulsed-field gel electrophoresis (PFGE) was used to determine the genomic fingerprints of Camospylobacter coli (n=27), Campylobacter jejuni (n=188), Arcobacter butzleri (n=138), Arcobacter cryaerophilus 1A (n=4), and A. cryaerophilus 1B (n=31) with the restriction enzymes SmaI and KpnI for Campylobacter and Arcobacter, respectively. Campylobacter species were subtyped by the Centers for Disease Control and Prevention PulseNet 24-h standardized protocol for C. jejuni. A modification of this protocol with a different restriction endonuclease (KpnI) and different electrophoresis running conditions produced the best separation of restriction fragment patterns for Arcobacter species. Both unique and common PFGE types of Arcobacter and Campylobacter strains were identified. A total of 32.8% (57 of 174) of the Arcobacter isolates had unique PFGE profiles, whereas only 2.3% (5 of 215) of the Campylobacter isolates belonged to this category. The remaining Arcobacter strains were distributed among 25 common PFGE types; only eight common Campylobacter PFGE types were observed. Cluster analysis showed no associations among the common PFGE types for either genus. Each of the eight common Campylobacter types consisted entirely of isolates from one sampling day, whereas more than half of the common Arcobacter types contained isolates from different sampling days. Our results demonstrate far greater genetic diversity for Arcobacter than for Campylobacter and suggest that the Campylobacter types are specific to individual flocks of birds processed on each sampling day.

Animals↗

Detection and diversity of various Arcobacter species in Danish poultry.

The prevalence and diversity of different Arcobacter spp. in various poultry species in Denmark were investigated using cultural and multiplex PCR methods. A pool of three fresh droppings obtained at the production site from 70 broiler chicken flocks aged 4-5 weeks was examined. In addition, pools of 10 cloacal swabs taken at the abattoir prior to stunning from each of 15, and 37 duck and turkey flocks, respectively, were analyzed. Thirty fresh broiler chicken carcasses and 29 cloacal swabs from the respective viscera were also examined at the abattoir. Finally, 10 caecal and 10 cloacal swabs from ducks at the abattoir were analyzed individually. In total, 85 Arcobacter isolates were obtained. Of these 45, 20 and 7 were identified as Arcobacter butzleri, Arcobacter cryaerophilus and Arcobacter skirrowii, respectively, using a multiplex PCR. Interestingly, some chicken isolates of A. butzleri showed urease activity, and 6 out of seven A. skirrowi isolates were unable to hydrolyse indoxyl acetate. All chicken carcasses examined were found positive for A. butzleri and/or A. cryaerophilus, whereas 21 (72%) of the 29 chicken cloacal swabs were positive for either A. butzleri (13) or A. cryaerophilus (9). Three (4.3%) out of 70 chicken flocks analyzed were positive only for A. cryaerophilus. Of the ten ducks examined individually, 7 carried A. skirrowii and/or A. cryaerophilus in their cloacae. None of the respective caecal samples were positive. Of the remaining 15 duck flocks, 11 (73%) were positive for A. cryaerophilus (7), A. butzleri (2) or A. skirrowii (2). Four (11%) of the 37 turkey flocks analyzed harboured either A. butzleri or A. cryaerophilus. The carriage rate of Arcobacter was higher in live ducks than those of live broiler chickens and turkeys in the present study. In addition, chicken carcasses slaughtered in Denmark were found to be contaminated with Arcobacter. The presence of Arcobacter spp. both on chicken carcasses and in poultry intestine may be of significance to human health.

Abattoirs↗

Prevalence and diversity of Arcobacter spp. isolated from the internal organs of spontaneous porcine abortions in Denmark.

A study was conducted to determine the prevalence and possible significance of campylobacteria in pig abortions in Denmark. Surface-cauterised liver and kidney samples from 55 aborted pig fetuses submitted to the Danish Veterinary Laboratory were taken and a sensitive isolation procedure used to examine pooled tissue samples for Campylobacter, Arcobacter and Helicobacter spp. Routine microbiological, immunological, and histopathological examinations were also performed to identify concurrent infections or histopathological changes. The abortions tested negative for established abortifacient pathogens (Brucella, Leptospira, PPV, PRRSV), but Arcobacter spp. were recovered from 23/55 abortions. Co-infections with Streptococcus suis, Escherichia coli, and haemolytic streptococci were observed in 7/23 Arcobacter-positive fetuses, and in 4/32 Arcobacter-negative fetuses. Histopathological analyses identified placentitis, pneumonia, hepatitis and encephalitis among the study group. However, no obvious pathologic features were solely associated with Arcobacter-positive cases, nor were Arcobacter-like bacteria observed in tissue samples. Protein profile analyses of the 27 Arcobacter isolates identified 11 as A. cryaerophilus and 10 as A. skirrowii. Six strains could not be classified into any existing species and were phenotypically distinct, thus, potentially representing at least one new species. The identification results showed that multiple taxa could be found in a single fetus, and in distinct aborted fetuses from a single sow. The high prevalence of arcobacters in Danish pig abortions may account for at least some of the >90% of cases in which no established abortifacient agent is detected, but further studies are needed to define the role of each species, especially where co-infections with other bacteria are present.

Abortion, Veterinary↗

Detection of Arcobacter spp. in the coastal environment of the Mediterranean Sea.

The occurrence of Arcobacter spp. was studied in seawater and plankton samples collected from the Straits of Messina, Italy, during an annual period of observation by using cultural and molecular techniques. A PCR assay with three pairs of primers targeting the 16S and 23S rRNA genes was used for detection and identification of Arcobacter butzleri, Arcobacter cryaerophilus, and Arcobacter skirrowii in cultures and environmental samples. Only one of the Arcobacter species, A. butzleri, was isolated from seawater and plankton samples. With some samples the A. butzleri PCR assay gave amplified products when cultures were negative. A. cryaerophilus and A. skirrowii were never detected by culture on selective agar plates; they were detected only by PCR performed directly with environmental samples. Collectively, our data suggest that culturable and nonculturable forms of Arcobacter are present in marine environments. The assay was useful for detecting Arcobacter spp. both as free forms and intimately associated with plankton. This is the first report showing both direct isolation of A. butzleri and the presence of nonculturable Arcobacter spp. in the coastal environment of the Mediterranean Sea.

Animals↗

Isolation of Arcobacter spp. from retail meats and cytotoxic effects of isolates against vero cells.

A survey of Arcobacter spp. was conducted over a 12-month period in Guadalajara, Mexico. A total of 135 samples (45 lean ground beef samples, 45 lean ground pork samples, and 45 chicken samples, including drumsticks, gizzards, and ground or chopped breast) were collected from local butcheries. The samples were enriched in Johnson-Murano enrichment medium and then streaked onto Johnson-Murano agar plates. Typical colonies were subjected to microscopic and biochemical identification followed by polymerase chain reaction confirmation of the genus Arcobacter. All isolates confirmed to be Arcobacter isolates were then inoculated into Eagle's minimum essential medium to determine their cytotoxicity against Vero cells. Arcobacter spp. were detected in 28.8, 51.1, and 40.0% of beef, pork, and chicken samples, respectively. From these samples, 101 isolates were confirmed to be Arcobacter spp. by polymerase chain reaction. Overall, the species most frequently identified was A. butzleri, followed by A. skirrowii. A. cryaerophilus was isolated only from pork meat. Ninety-five (95%) of the Arcobacter isolates produced a virulence mechanism against Vero cells, and 38 of them induced cell elongation, indicating enterotoxin production. Eighteen isolates produced the formation of vacuoles, and 39 produced both vacuolization and elongation. The vacuolization effect may be related to a vacuolizing toxin. The production of a vacuolizing toxin by Arcobacter spp. has not previously been reported. Results obtained in this study indicate that Arcobacter spp. may show cytotoxic effects other than the recognized enterotoxin production.

Animals↗

Prevalence of Arcobacter and Campylobacter on broiler carcasses during processing.

Broiler carcasses (n=325) were sampled in a U.S. commercial poultry processing plant for the prevalence of Arcobacter and Campylobacter at three sites along the processing line: pre-scald, pre-chill and post-chill. Samples (75-125 broilers per site) were collected during five plant visits from August to October of 2004. Arcobacter was recovered from pre-scald carcasses more frequently (96.8%) than from pre-chill (61.3%) and post-chill carcasses (9.6%). Campylobacter was isolated from 92% of pre-scald carcasses, 100% of pre-chill carcasses, and 52% of post-chill carcasses. In total, Arcobacter was isolated from 55.1% (179 of 325), while Campylobacter was isolated from 78.5% (255 of 325) of the carcasses from the three collection sites. For Arcobacter identification, a species-specific multiplex PCR showed that A. butzleri was the most prevalent species (79.1%) followed by A. cryaerophilus 1B (18.6%). A. cryaerophilus 1A was found at low levels (2.3%). PCR identified the most common Campylobacter species as C. jejuni (87.6%) followed by C. coli (12.4%). Overall, significant contamination of broiler carcasses by Arcobacter was observed, although less than that found for Campylobacter. From pre-scald to post-chill, a far greater reduction in Arcobacter numbers was observed than for Campylobacter. Our results for Arcobacter, obtained from the same environment as the closely related pathogen Campylobacter, will aid in the development of control measures for this emerging pathogen.

Animals↗

Isolation and characterisation of Arcobacter butzleri from meat.

A survey was conducted to determine the prevalence of Arcobacter in ground chicken, pork, beef and lamb meats. Meat samples were enriched in Arcobacter broth (AB) containing cefoperazone, amphotericin and teicoplanin (CAT) supplement. Samples were screened for the presence of Arcobacter spp. using a multiplex polymerase chain reaction (PCR) followed by isolation on blood and selective agar. Arcobacter butzleri was the only species of Arcobacter isolated from 35% of 88 samples of ground meats. A. butzleri was more frequently isolated from poultry (73%) than pork (29%), beef (22%) or lamb (15%) samples. No significant differences were found in the isolation rates and from the different regions sampled. Isolates were characterised by pulsed-field gel electrophoresis (PFGE) using SacII, EagI and SmaI restriction endonucleases. A number of isolates with indistinguishable PFGE fingerprints were found to be epidemiologically related, which may indicate cross-contamination of common types of Arcobacter from different meat species or between meat species. The public health significance of Arcobacter in ground meat needs to be determined.

Animals↗

Ecology of Arcobacter species in chicken rearing and processing.

AIMS: To investigate whether Arcobacter spp. colonize the poultry-rearing environment or whether they are contaminants acquired during transportation and/or from the processing plant. METHODS AND RESULTS: Samples were collected on poultry farms and in the processing plant during slaughter and dressing. Two cultural methods of detection were used. Isolates were identified to species level using a multiplex-polymerase chain reaction (m-PCR) method, either on the initial suspensions, or after enrichment, or on pure cultures of isolates. Of the 62 samples examined from poultry farms, arcobacters were found only outside the rearing sheds (in effluent sludge and stagnant water). Thirty-four samples were examined from the processing plant and 26 were positive for arcobacters. All the isolates were Arcobacter butzleri. Arcobacters were not found in any sample by direct plating nor by m-PCR on the initial suspensions, thus it was concluded that numbers were very low. CONCLUSIONS: Arcobacter spp. were not found in samples from the live birds and their immediate environment, but A. butzleri was found in effluent sludge and stagnant water outside the rearing sheds. However, A. butzleri is common in poultry abattoirs, and it appears that poultry carcasses are contaminated during processing. SIGNIFICANCE AND IMPACT OF THE STUDY: Arcobacters are not found inside poultry-rearing sheds, but are contaminants in the processing environment.

Abattoirs↗

Identification by 16S ribosomal RNA gene sequencing of Arcobacter butzleri bacteraemia in a patient with acute gangrenous appendicitis.

AIMS: To identify a strain of Gram negative facultative anaerobic curved bacillus, concomitantly isolated with Escherichia coli and Streptococcus milleri, from the blood culture of a 69 year old woman with acute gangrenous appendicitis. The literature on arcobacter bacteraemia and arcobacter infections associated with appendicitis was reviewed. METHODS: The isolate was phenotypically investigated by standard biochemical methods using conventional biochemical tests. Genotypically, the 16S ribosomal RNA (rRNA) gene of the bacterium was amplified by the polymerase chain reaction (PCR) and sequenced. The sequence of the PCR product was compared with known 16S rRNA gene sequences in the GenBank by multiple sequence alignment. Literature review was performed by MEDLINE search (1966-2000). RESULTS: The bacterium grew on blood agar, chocolate agar, and MacConkey agar to sizes of 1 mm in diameter after 24 hours of incubation at 37 degrees C in 5% CO2. It grew at 15 degrees C, 25 degrees C, and 37 degrees C; it also grew in a microaerophilic environment, and was cytochrome oxidase positive and motile, typically a member of the genus arcobacter. Furthermore, phenotypic testing showed that the biochemical profile of the isolate did not fit into the pattern of any of the known arcobacter species. 16S rRNA gene sequencing showed one to two base differences between the isolate and A butzleri, but 35 to 39 base differences between the isolate and A cryaerophilus, indicating that the isolate was a strain of A butzleri. Only three cases of arcobacter bacteraemia with detailed clinical characteristics were found in the English literature. The sources of the arcobacter species in the three cases were largely unknown, although the gastrointestinal tract is probably the portal of entry of the A butzleri isolated from the present case because the two concomitant isolates (E coli and S milleri) in the blood culture were common flora of the gastrointestinal tract. In addition, A butzleri has previously been isolated from the abdominal contents or peritoneal fluid of three patients with acute appendicitis. CONCLUSIONS: 16S rRNA gene sequencing was useful in the identification of the strain of A butzleri isolated from the blood culture of a patient with acute gangrenous appendicitis. Arcobacter bacteraemia is rare. Further studies using selective medium for the delineation of the association between A butzleri and acute appendicitis are warranted.

Acute Disease↗

Prevalence of Arcobacter spp. in raw ground pork from several geographical regions according to various isolation methods.

Studies show that the pathogen Arcobacter is present in beef, poultry, and pork. Several methods have been reported for the isolation of this organism, but none has been adopted as the standard. This has limited the significance of field comparison studies. In the present study, we compared the efficiencies of four Arcobacter isolation methods using raw ground pork collected from slaughter facilities across the United States. We also evaluated the effect of meat fat level and age of animals on the prevalence of Arcobacter in ground pork. The methods chosen for comparison of isolation efficiency were those of Collins, a modified version of the Collins method (Direct Collins), deBoer, and Johnson Murano (JM). These were chosen based on published reports in which they were used to detect Arcobacter in pork products. The JM method was found to be the most successful in consistently detecting Arcobacter, isolating it in 64 of 200 pork samples compared with the Direct Collins method, which isolated Arcobacter in 52 of 200 of those same samples. The Collins method and the deBoer method found Arcobacter present in only a fraction of the samples. The level of contamination was found to vary among the plants, ranging from 0% to 68% prevalence, with 32% overall for all four plants tested. Additionally, ground pork low in fat had a higher contamination frequency (20%) when compared with high-fat pork (4%). Results also showed that meat from younger animals was more frequently contaminated than that from older animals.

Age Factors↗

Lack of a cytolethal distending toxin among Arcobacter isolates from various sources.

Arcobacter has been shown to be present in numerous different sources, including poultry, water, and humans exhibiting gastroenteritis. The production of a cytolethal distending toxin (CDT) has been documented in Campylobacter, Helicobacter, and other species. The polymerase chain reaction was used to screen Arcobacter isolates from poultry, cattle, irrigation water, and human diarrhea for the presence of CDT genes. Cell filtrates and sonic extracts were also tested for CDT-like activity on Chinese Hamster Ovary, HeLa, and Intestinal 407 (INT407) cells in culture. No CDT amplimers were observed in any of the Arcobacter isolates investigated. However, toxicity to HeLa and INT407 cells was observed and was subsequently analyzed for cell cycle arrest in the presence of the Arcobacter extracts with flow cytometry. Cells treated with Arcobacter sonic extracts and filtrates exhibited normal cell cycles, suggesting that CDT is not expressed by Arcobacter. Thus, Arcobacter was shown to produce an entity that was toxic to some cells in culture, but this entity was toxic in a manner different from that of Campylobacter CDT.

Animals↗

Molecular characterization of Arcobacter isolates collected in a poultry slaughterhouse.

In a poultry slaughterhouse, Arcobacter contamination was examined over a period of 1 week to establish possible routes of contamination. Samples were collected from the slaughter equipment and from processing water before the onset of slaughter and from the first broiler flock slaughtered on each sampling day. Characterization of 1,079 isolates by enterobacterial repetitive intergenic consensus-polymerase chain reaction and a random amplified polymorphic DNA assay resulted in the delineation of 159 Arcobacter butzleri and 139 Arcobacter cryaerophilus types. From almost all 140 neck skin samples collected before and after evisceration, A. butzleri and A. cryaerophilus were isolated simultaneously at contamination levels ranging from 10(1) to 10(4) CFU/g. Only six A. butzleri types present in the slaughterhouse environment were also present on the broiler carcasses. None of the A. cryaerophilus genotypes were detected in both the neck skin and the environmental samples. All A. butzleri types isolated from the feather samples were also isolated from broiler neck skin samples. The slaughter equipment was contaminated with arcobacters before the onset of slaughter, but it appeared unlikely that contamination through the slaughter equipment alone explained the high contamination levels on poultry products. Arcobacters were also present in processing water, but types present in water and poultry products were different. Characterization of the Arcobacter isolates did not clarify the routes of transmission, probably because of the extreme heterogeneity among Arcobacter isolates. However, the results obtained in this study brought to light insufficient decontamination at the processing plant involved in the study and confirmed the survival capacity of certain A. butzleri strains.

Abattoirs↗

A comparison of three methods for the isolation of Arcobacter spp. from retail raw poultry in Northern Ireland.

Recent evidence suggests that arcobacters, especially Arcobacter butzleri, are potential foodborne pathogens, but standardized detection methods have yet to be established. A study was undertaken to determine which of three isolation methods was the most effective for the isolation of Arcobacter spp. from fresh raw poultry. Methods 1 was microaerobic and involved a membrane filtration step followed by plating onto blood agar. Method 2 was also microaerobic and involved enrichment and plating media containing a five-antibiotic cocktail. Method 3 was aerobic and was based on enrichment in a charcoal-based broth containing two antibiotics. Retail poultry samples (n = 50) were obtained from supermarkets in Northern Ireland; the European Community license number was recorded to ensure sample diversity. Presumptive arcobacters were identified using genus-specific and species-specific primers. Methods 1 resulted in the lowest recovery of arcobacters (28% of samples positive). The detection rate for method 2 (68%) was higher than that for method 3 (50%), but the difference was not significant (P > 0.05). Modification of method 3 by plating the enrichment broth at 24 h, as well as at 48 h, increased recovery to 68%. Use of methods 2 and 3 together increased the number of positive samples detected by approximately 25% compared with use of either method alone. A. butzleri was the most commonly isolated species using all methods. Method 3 detected Arcobacter cryaerophilus in more samples (n = 3) than did method 1 and 2 (n = 1). Arcobacter skirrowii was detected by only method 3 (n = 1). In terms of sensitivity, ease of use, and diversity of species recovered, modified method 3 was the overall method of choice.

Animals↗

Prevalence of Arcobacter spp. in raw milk and retail raw meats in Northern Ireland.

A 1-year study was undertaken to determine the prevalence of Arcobacter spp. in raw milk and retail raw meats on sale in Northern Ireland. Retail raw poultry samples (n = 94), pork samples (n = 101), and beef samples (n = 108) were obtained from supermarkets in Northern Ireland, and raw milk samples (n = 101) were kindly provided by the Milk Research Laboratory, Department of Agriculture and Rural Development, Belfast, Northern Ireland. Presumptive arcobacters were identified by previously described genus-specific and species-specific PCR assays. Arcobacter spp. were found to be common contaminants of retail raw meats and raw milk in Northern Ireland. Poultry meat (62%) had the highest prevalence, but frequent isolations were made from pork (35%), beef (34%), and raw milk (46%). Arcobacter butzleri was the predominant species isolated from retail raw meats and was the only species isolated from raw milk samples. Arcobacter cryaerophilus was detected less frequently, and Arcobacter skirrowii was detected only as a cocontaminant. To our knowledge, this is the first report of Arcobacter spp. prevalence in a diverse range of products of animal origin in Northern Ireland.

Animals↗

Evaluation of a new arcobacter enrichment medium and comparison with two media developed for enrichment of Campylobacter spp.

The productivity of an arcobacter enrichment medium (AM), newly developed by Oxoid was compared with two campylobacter enrichment media (Preston broth (Oxoid) and LabM broth), with arcobacter basal medium (ABM) as control. Twenty strains of Arcobacter and Campylobacter spp. were tested for growth, with target inocula of < 4 cfu per ml of medium. Incubation was carried out aerobically for 48 h with tightly closed caps at 25 degrees C for arcobacters and 37 degrees C for campylobacters. After incubation the numbers of cfu in the broths were counted by surface-plating on blood agar. None of the Campylobacter spp. grew in the complete AM, and only one grew (very poorly) in the arcobacter basal medium. However, AM supported good growth of all three species of Arcobacter (A. butzleri, A. skirrowii and A. cryaerophilus) which have been associated with human and animal disease. The one strain of A. nitrofigilis tested failed to grow in any of the media except poorly in ABM. None of the Arcobacter spp. grew in Preston Broth, but nine grew in LabM broth, although productivity was poor compared to AM. None of the Campylobacter spp. grew in AM and only one grew (very poorly) in ABM.

Campylobacter↗