API Campy Research Articles

Современные методы видовой идентификации термофильных бактерий рода Campylobacter

Emergent thermophilic Campylobacter hepaticus is the causative agent of Spotty Liver Disease (SLD) in laying hens. C. hepaticus is difficult to cultivate because commercial media for the isolation and cultivation of Campylobacter contain cefoperazone, which inhibits many isolates of the C. hepaticus species. Campylobacter was isolated using modified Preston broth, incubated at 37 °C under microaerophilic conditions for 7 days and then subcultured onto selective Preston agar, erythritol agar with Oxoid selective additives and 5–7% defibrinated horse blood. Commercial test systems (API Campy) were used for identification. The use of the classical bacteriological diagnostic method, which is considered the 'gold' standard, is limited due to the difficulties of cultivation. The identification of new Campylobacter species requires revision of phenotypic identification algorithms. Specific primers for the identification of new Campylobacter species also need to be developed. In our studies, using the KAM-BAC kit, we detected Campylobacter jejuni DNA in clinically healthy birds. Consequently, the carriage of Campylobacter is massive. 30 samples of test material were examined using the molecular-biological method, and 60 samples using the bacteriological method. Analyzing the results of Campylobacter detection, it should be noted that thermophilic Campylobacteria were isolated from 60 clinical samples by the bacteriological method in 5,0% (3 Campylobacter cultures), and from 30 samples by the molecular-biological method in 27,0% (8 positive samples). Based on the analysis of the study results, it is necessary to conduct an in-depth study of the natural sources of Campylobacter hepaticus distribution, virulence factors, pathogenesis and mechanisms of infections caused by these emergent pathogens. The most promising research in the study of the causative agents of Campylobacteriosis in birds will be based on the application of innovative genomic technologies based on multiplex polymerase chain reactions and genome sequencing of Campylobacter hepaticus.

Prevalence of <i>Campylobacter</i> spp. Isolated from Pigs in Three Slaughterhouses of Yaounde Cameroon

Enteric campylobacteriosis caused by Campylobacter is a real public health problem. It is a typical zoonosis that can be transmitted through direct contact with contaminated animals or animal carcasses, or indirectly through the ingestion of contaminated food or water. WHO1 defines Campylobacter as the first germ responsible for gastroenteritis in developing countries. The objective of the study is to determine the prevalence of Campylobacter in pigs and to identify the most common species found in three slaughterhouses in the city of Yaounde. A descriptive and prospective study was carried out over a period of five months, from July to November 2020. To do this, Campylobacter spp. strains were searched for by culture on specific media, incubated in a micro-aerophilic atmosphere at 37°C and the identification with API campy (BioMerieux, ref 20800). Out of the 266 samples collected, 170 from the rectum and 96 from the carcasses, we obtained a general prevalence of 27%, with a prevalence of 25.88% in the rectum and 29.16% in the carcasses. Pigs intended for consumption by the population of Yaounde come mainly from the North (50%) and the West (49%). The most common species encountered were C. coli (52.77%), C. jejuni spp. jejuni 1 and 2 (30.58%), C. festus spp. festus (11.11%), C. jejuni spp. doley (1.38%), and Campylobacter spp. (4.16%). Bacterial enumeration gave Campylobacter fecal concentrations at 103 CFU2 (52.7%), 104 CFU (33.3%), and 105 CFU (13.8%). The slaughterhouse in the Mvog Ada market was the most involved in Campybacter cases, followed by the 8th market and Essos market. From the above, it is important to notice that the Campylobacter danger remains and that preventive measures are of great interest.

Salmonella spp. and Campylobacter spp. in poultry feces and carcasses in Ouagadougou, Burkina Faso.

The importance of Salmonella and Campylobacter as foodborne pathogens is well recognized worldwide. Poultry and poultry products are commonly considered as the major vehicles of Salmonella and Campylobacter infection in humans. The aim of this study was to investigate the hygienic status of poultry facilities and determine the prevalence of Salmonella and Campylobacter in slaughtered poultry feces and carcasses in four different markets in Ouagadougou, capital city of Burkina Faso. A total of 103 poultry feces and 20 carcasses were analyzed using microbiological standard methods. Among the 103 fecal samples, 70 were positive for Campylobacter ssp (67.96%) and 54 for Salmonella ssp (52.42%). The hippurate hydrolysis test revealed that among the 70 Campylobacter strains isolated from feces, 49 were C. jejuni (70%) and 21 were C. coli (30%). From the 20 carcasses analyzed, 18 were contaminated with Salmonella (90%) and 10 with Campylobacter ssp (50%). Among the 10 Campylobacter ssp samples isolated from poultry carcasses, all were identified as C. jejuni using the API CAMPY system and the hippurate hydrolysis test. The assessment of markets hygienic practices for production, transportation, display, and vending of meat revealed unhygienic conditions. To complete the observation of unhygienic practices, we have sampled chicken‐washing solution from the study sites and microbiological analysis of these samples revealed the presence of Salmonella spp in 100% of the samples. This study highlighted that poultry products on sale in Ouagadougou are highly contaminated with Salmonella and Campylobacter. To the best of our knowledge, this is the first report describing Campylobacter presence in the poultry industry of Burkina Faso. Our findings might help to better understand the epidemiology of Salmonella and Campylobacter.

English

Poultry meat and products are major transmission routes of human campylobacteriosis. The aim of this study was to determine the numbers and antibiogram profile of Campylobacter isolates from slaughtered broiler and layer birds. One hundred and sixty caecal and one hundred and thirty two carcasses were randomly sampled at the Kejetia poultry slaughter, isolated on charcoal-cefoperazone-deoxycholate agar (CCDA) and confirmed by API CAMPY and their resistance profiles assessed by Kirby-Bauer disk diffusion. Prevalence was 22.5 and 21.9% in the faecal and carcasses, respectively with no significant differences. Species identified among faecal isolates were Campylobacter jejuni (42%), Campylobacter coli (28%), Campylobacter lari (22%) and Campylobacter hyo-intestinalis (8%) while 79% C. jejuni, 14% C. coli, 4% C. jejuni sub sp. doylei, and 3% C. lari were obtained from the carcasses. Resistance to the β-lactams ranged from 75 to 100%, 41 to 86% to the quinolones, 14 to 36% to the aminoglycosides, 100% to erythromycin, 97 to 100% to tetracycline, 72 to 83% to chloramphenicol and 90 to 94% to trimethoprim sulfamethoxazole. All species were sensitive to imipenem, but 100% of isolates were multidrug resistant. Contamination of carcasses with multidrug resistant strains of Campylobacter is a threat to handlers and consumers and of major public health issue. Key words: Multidrug resistance, faeces, carcass, poultry, Kumasi, Ghana.

Antibiotic Resistance of Campylobacter Recovered from Faeces and Carcasses of Healthy Livestock.

Campylobacter is of major significance in food safety and human and veterinary medicine. This study highlighted resistance situation in the area of veterinary public health in Ghana. Using selective mCCDA agar, isolates were confirmed phenotypically on API CAMPY and genotypically by multiplex PCR of IpxA gene. The susceptibility profile of species to common and relevant antibiotics was determined by the Kirby-Bauer disk diffusion method. Cattle, sheep, goat, and pig faecal samples analysed, respectively, yielded 13.2% (16/121), 18.6% (22/102), 18.5% (25/135), and 28.7% (29/101) Campylobacter species while 34.5% (38/110), 35.9% (42/117), 23.9% (32/134), and 36.3% (37/102) were, respectively, recovered from the carcasses. Species identified in faeces were C. jejuni 35.8% (33/92), C. jejuni subsp. doylei 4.3% (4/92), C. coli 47.8% (44/92), and C. lari 12.0% (11/92). Species discovered in carcasses were C. jejuni 83.9% (125/149), C. jejuni subsp. doylei 2.0% (3/149), C. coli 6.0% (9/149), and C. lari 8.1% (12/149). Resistance ranged from 92 to 97% to the β-lactams, 7 to 69% to the quinolones, 0 to 44% to the aminoglycosides, 97 to 100% to erythromycin, 48 to 94% to tetracycline, 45 to 88% to chloramphenicol, and 42 to 86% to trimethoprim/sulfamethoxazole as 0% resistance was observed against imipenem.

An important agent in gastroenteritis: Campylobacter

Campylobacter infections are common in both developed and developing countries. The reported incidence of culture confirmed infections shows variability among countries due to culturing procedures. (1). Here we report laboratory-based surveillance data collected between 2013 and 2015 in Yeditepe University Hospital Microbiology Laboratory. Among 2807 feces cultures, Salmonella spp. (n=135), Campylobacter spp. (n=118), Aeromonas spp. (n=6) and Shigella spp. (n=3), isolates were detected. Out of 118 Campylobacter spp. isolates, the distribution was as follows: C. jejuni (n=101), C. upsaliensis (n=8), C. coli (n=6) and other species (n=3). The fecal samples were cultured in Campylobacter-BAP medium (Salubris, Turkey) and incubated under microaerophilic conditions (CampyGen, Oxoid, UK) at 42°C for 48 hours. Suspected colonies were examined by Gram stain and evaluated for oxidase and catalase positivity; and then were first confirmed by Campylobacter latex agglutination test (Dryspot, Oxoid, UK). They were identified by API CAMPY biochemical identification system (BioMerieux, France). Antimicrobial susceptibility of isolates against erythromycin was also tested with API CAMPY system (BioMerieux, France); ciprofloxacin and gentamicin susceptibilies were tested by E-test (bioMerieux, France). The distribution of isolates, demographic information and seasonal epidemiology in patients with Campylobacter positive cultures are shown. It seems there is no definite season for campylobacteriosis. We mostly isolate from the pediatric patients. (Table 1, Figure 1) Beyond detecting difficulties by culture, antibiotic resistance becomes a problem. In the EFSA/ECDC report 2013, the highest frequency of resistance was observed for nalidixic acid (47.8%) and ciprofloxacin (44.4%) followed by ampicillin (35.3%) and tetracycline (30.5%); low resistance levels were observed to erythromycin (3.5%) and gentamicin (0.4%). According to FDA/CDC/UDSA Report 2013 resistance to tetracycline was most common (45.9%), followed by ciprofloxacin (23.5% in C. jejuni and 35.8% in C. coli). Macrolide resistance was observed in 1.7%-2.7% (2). During the study period, our resistance rates were detected as 13% for erythromycin, 11% for ciprofloxacin. It seems a slight increase in erythromycin resistance in our strains comparing to 2009-2012 rates (from 7% to 13%) (3). No resistance was seen in gentamicin. Fecal culture procedures should be applied in our routine laboratories to see the surveillance of Campylobacteriosis and find our national rate in resistance to antibiotics.

Antibiotic Resistance in <i>Campylobacter</i> Isolated from Patients with Gastroenteritis in a Teaching Hospital in Ghana

Campylobacter is a leading causal agent of bacterial enteritis worldwide, but its prevalence is not well documented in Ghanaian hospitals. This study isolated Campylobacter species from patients with enteritis or urinary tract infections attending Komfo Anokye Teaching Hospital and assessed the antibiogram profile of isolated species. Two hundred and two (202) in-patients and outpatients samples of all age groups diagnosed with enteritis or UTI infections were analyzed from May 2013 to August 2013. Campylobacter species were detected using selective agar (mCCDA) and confirmed on API Campy system (bioMerieux, France), with disk diffusion method determined the resistance profile of the species. Of the 128 enteritis and 74 UTI patients samples analyzed, 26 and 9 isolates were respectively confirmed as Campylobacter spp. giving a prevalence of 17.3% (35/202). Species identified were C. jejuni (40%), C. jejuni sub sp. doylei (2.8%), C. coli (37%) and C. lari (20%). Resistance was 92.3% - 100% each to erythromycin and the β-lactams, 61.5% - 86.7% to trimethoprim sulfamethoxazole, 92.3% - 93.3% to tetracycline, 46.2% - 80% to chloramphenicol, 0% - 60% to aminoglycosides and 0% to imipenem. Multidrug resistance of 97.1% was detected among species. Empirical treatment of Campylobacter enteritis with erythromycin and other common and cheap drugs may result in treatment failure in the face of high level resistance observed among the Campylobacter species.

English

Environmental waters are established sources of Campylobacter infections in humans. The aim of this study was to assess the distribution and susceptibility profiles of Campylobacter species isolated from irrigation and domestic water sources in Ghana. Samples were pre-enriched with CCDA broth and isolated on mCCDA agar. Isolates were confirmed on API CAMPY kit and with the Kirby-Bauer disk diffusion method to determine the susceptibility patterns of species. Of the 188 water samples analyzed, 42 isolates were confirmed to be contaminated with Campylobacter species, giving a prevalence rate of 22.3%. Prevalence of Campylobacter in the various water sources were 35.7% in rivers, 26.2% in streams, 21.4% in wells, 9.5% in ponds and 7.1% in boreholes. Sixty four percent (64%) of Campylobacter species were Campylobacter jejuni, followed by Campylobacter coli (19%) and Campylobacter lari (14%). Resistance was 100% to the β-lactams, 98% to erythromycin, 48-69% to the quinolones, 45-55% to the aminoglycosides, 71% to trimethoprim sulphamethoxazole, 76% to tetracycline and 90% to chloramphenicol. All the isolates (100%) were multidrug resistant. The presence of multidrug resistant Campylobacter species in the different water sources sampled in this study may indicate a significant health risk to humans and animals. Key words: Campylobacter, antibiotic resistance, water sources, Ghana.

Campylobacter jejuni ssp. jejuni bacteraemia in a patient with BCR-ABL-positive chronic myelogenous leukaemia in remission on dasatinib therapy

Introduction: We report an unusual case of Campylobacter jejuni bacteraemia in a patient with Philadelphia chromosome-positive, BCR-ABL-positive chronic myelogenous leukaemia in complete cytogenetic and major molecular remission after dasatinib/IFN therapy. Case presentation: On admission, the patient presented with fever and acute haemorrhagic diarrhoea. Initial empiric antibiotics consisted of ceftriaxone and metronidazole. Stool and blood culture samples were collected and submitted for evaluation; these specimens were processed as per laboratory protocol. Several Gram-negative, spiral rods could be identified microscopically in Gram-stained slides from bottled blood and stool cultures. Isolate identification was performed on the Vitek 2 system using a GN identification card and API Campy strips and by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). Discrepant identification was obtained between the Vitek versus the API Campy test and MALDI-TOF. Erythromycin susceptibility testing was done using an Etest, whereas all other susceptibility testing and breakpoint analysis was done following the EUCAST procedure. Initial empiric antibiotic treatment was switched from ceftriaxone to ciprofloxacin according to antibiotic susceptibility testing (AST) and Etest (ciprofloxacin MIC of 0.094 μg ml− 1) results. C. jejuni bacteraemia was successfully eradicated after 1 week of ciprofloxacin therapy. Conclusion: This case describes the rare event of a Campylobacter bacteraemia, which could have been falsely interpreted because of non-specific colony morphology and the initial Vitek result. Only the combination of microscopy, MALDI-TOF and an API Campy test was able to rapidly identify this bacterium as C. jejuni. This case also shows the importance of immediate AST- and MIC-adapted antibiotic treatment in immunocompromised febrile patients suffering from Campylobacter-induced bloody diarrhoea.

Prevalence of Campylobacter jejuni in chicken produced by major poultry companies in Saudi Arabia

Abstract Background Campylobacter is a foodborne pathogen that is commonly associated with chicken. The aim of this work was to evaluate the prevalence of Campylobacter jejuni (as affected by refrigerated storage) in chicken samples obtained from the wholesale poultry market in the northern part of Riyadh City, Saudi Arabia. Findings A gradual increase in the number of positive samples was noted during storage at 4°C. On days 1, 3, and 7, the number of positive samples were 10 (30.305%), 15 (45.45%), and 27 (81.81%), respectively. Of 99 tested samples, 52 (52.25%) were positive for Campylobacter jejuni. Protein profiling by Sodium dodecyl sulfate -Polyacrylamide gel electrophoresis (SDS-PAGE) was used to identify Campylobacter jejuni. The results were verified using Analytical Profile Index (API Campy system, Marcy l’Etoile, France). Forty-three (82.69%) positive isolates were identified as C. jejuni subsp. jejuni 2, 5 isolates as C. jejuni subsp. jejuni 1 (9.61%), and 4 isolates as C. jejuni subsp. doylei (7.69). Conclusion C. jejuni positive samples increased rapidly during storage at 4°C for approximately 1 wk. Our results also indicated a connection between the protein profiles on SDS-PAGE and API Campy used for the identification of C. jejuni.

Identification and Antimicrobial Resistance of Campylobacter Species Isolated from Animal Sources

Background: Campylobacter spp. are together with Salmonella spp. the leading causes of human bacterial gastroenteritis worldwide. The most commonly isolated species in humans are Campylobacter jejuni and C. coli. The isolation, identification, and antimicrobial resistance of Campylobacter spp. from poultry and raw meat from slaughterhouses, has been investigated for the first time in Greece. During the period from August 2005 to November 2008 a total of 1080 samples were collected: (a) 830 fecal samples from five poultry farms, (b) 150 cecal samples from chicken carcasses in a slaughterhouse, and (c) 100 fecal samples from one pig farm near the region of Attica. The identification of the isolates was performed with conventional (sodium hippurate hydrolysis and commercial identification system (Api CAMPY system, bioMerieux, France), as well as with and molecular methods based on 16S rRNA species specific gene amplification by PCR and subsequent sequence analysis of the PCR products. Results: Sixteen Campylobacter strains were isolated, all collected from the poultry farms. None of the strains was identified as C. jejuni. Antimicrobial susceptibility to six antimicrobials was performed and all the strains were susceptible to ciprofloxacin, amoxicillin–clavulanic acid, and gentamicin. Thirteen out of 14 C. coli were resistant to erythromycin and all C. coli strains were resistant to ampicillin. Conclusion: Our results emphasize the need for a surveillance and monitoring system with respect to the prevalence and antimicrobial resistance of Campylobacter in poultry, as well as for the use of antimicrobials in veterinary medicine in Greece.

Accuracy of the API Campy system, the Vitek 2 Neisseria–Haemophilus card and matrix-assisted laser desorption ionization time-of-flight mass spectrometry for the identification of Campylobacter and related organisms

Biochemical identification of Campylobacter and related organisms is not always specific, and may lead to diagnostic errors. The API Campy, the Vitek 2 system and matrix-assisted desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) are commercially available methods that are routinely used for the identification of these microorganisms. In the present study, we used 224 clinical isolates and ten reference strains previously identified by multiple PCR assays, whole cell protein profiling and either DNA–DNA hybridization or sequencing analysis to compare the reliability of these three methods for the identification of Campylobacter and related pathogens. The API Campy accurately identified 94.4% of Campylobacter jejuni ssp. jejuni and 73.8% of Campylobacter coli, but failed to correctly identify 52.3% of other Epsilobacteria. The Vitek 2 Neisseria–Haemophilus card correctly identified most C. jejuni ssp. jejuni (89.6%) and C. coli (87.7%) strains, which account for the majority of campylobacterioses reported in humans, but it failed in the identification of all of the other species. Despite a good identification rate for both C. jejuni ssp. jejuni and C. coli, both methods showed poor sensitivity in the identification of related organisms, and additional tests were frequently needed. In contrast to API Campy and Vitek, MALDI-TOF MS correctly identified 100% of C. coli and C. jejuni strains tested. With an overall sensitivity of 98.3% and a short response time, this technology appears to be a reliable and promising method for the routine identification of Campylobacter and other Epsilobacteria.

Prevalence, strain identification and antimicrobial resistance of Campylobacter spp. isolated from slaughtered pig carcasses in Brazil

The aim of this study was to investigate the rate of contamination, species identification and antimicrobial resistance of thermophilic Campylobacter spp. in pig carcasses during the slaughter process in a slaughterhouse in Brazil. Two hundred and fifty-nine samples were collected at 7 different stages of the slaughter process for Campylobacter determination by both qualitative and quantitative methods. Typical colonies were subjected to API Campy, real-time polymerase chain reaction (PCR) and antimicrobial resistance testing. Campylobacter was found in 18.9% of the carcasses and 3.5% of the samples. Dehairing was the slaughter stage with the highest Campylobacter contamination (55.6%). All Campylobacter strains were confirmed by real-time PCR and showed multi-drug resistance to cephalothin, nalidixic acid, norfloxacin, tetracycline and trimethoprim. None of the strains were resistant to amoxicillin/clavulanic acid, ampicillin and chloramphenicol. Despite the low occurrence of Campylobacter spp. in pig samples, the antimicrobial resistance of Campylobacter strains represents a considerable risk for the consumption of pork meat and confirms the need for continuous monitoring of Campylobacter in the pig production chain.

Morphologic, Genetic, and Biochemical Characterization of Helicobacter Magdeburgensis, a Novel Species Isolated from the Intestine of Laboratory Mice

The presence of enterohepatic Helicobacter species (EHS) is commonly noted in mouse colonies. These infections often remain unrecognized but can cause severe health complications or more subtle host immune perturbations and therefore can confound the results of animal experiments. The aim of this study was to isolate and characterize a putative novel EHS that has previously been detected by PCR screening of specific-pathogen-free mice. Biochemical analysis of enzyme activities (API campy), morphologic investigation (Gram-staining and electron microscopy) and genetic analyses (16SrRNA and 23SrRNA analyses, DNA fingerprinting, restriction fragment polymorphisms, and pulsed-field gel electrophoresis) were used to characterize isolated EHS. Genomic DNA fragments were sequenced to develop a species-specific PCR detection assay. Scanning electron microscopy revealed the presence of spiral-shaped EHS, which varied in length (2.5-6 μm) and contained single monopolar or single bipolar sheathed flagella. The bacteria were grown under anaerobic conditions, preferably on agar plates containing serum or blood. The 16SrRNA, genetic, and biochemical analyses indicated the identification of a novel EHS species, named Helicobacter magdeburgensis. We also examined the genome content using pulsed-field gel electrophoresis. Based on the pattern produced by two restriction enzymes, BamIII and KspI, the genome size was determined to be about 1.7-1.8 Mbp. We isolated and characterized a novel EHS species, H. magdeburgensis, morphologically, biochemically, and genetically. These results are important for future studies on the prevalence and pathophysiologic relevance of such infections. Our PCR assay can be used to detect and discriminate H. magdeburgensis from other Helicobacter species.

Occurrence and Characterization of Campylobacter in the Brazilian Production and Processing of Broilers

Twenty-two commercial broiler flocks and their carcasses, totaling 546 samples (450 collected from a poultry farm and 96 from a slaughterhouse), were surveyed for the presence of Campylobacter. The positive results for Campylobacter among the analyzed samples were homogeneous, yielding 81.8% for cecal droppings, 80.9% for feces, and 80.4% for cloacal swabs. Pre-enrichment and direct plating showed that 77.85% and 81.8% of cloacal swabs, respectively, were positive for Campylobacter compared to 99.0% and 97.9% of carcasses testing positive with the pre-enrichment and direct plating methods. The Campylobacter count averaged 7.0 log10 colony-forming units (CFU)/g in cecal droppings, 5.15 log10 CFU/carcass after defeathering, and 4.24 log10 CFU/carcass after chilling. The samples were identified by the API Campy system as Campylobacter jejuni subsp. jejuni (68.8%), Campylobacter coli (8.3%), Campylobacter jejuni subsp. doylei (6.3%), Campylobacter upsaliensis (4.2%), and Campylobacter fetus subsp. fetus (2.1%). The analyzed broiler flocks were positive for Campylobacter in 81.8% of the cases, thus characterizing the occurrence of this pathogen in a broiler-producing region in southern Brazil. These results highlight the importance of programs targeted at the reduction of Campylobacter in poultry products, in order to minimize the risks for consumers.

Presence of thermophilic Campylobacter species in Broilers and pigs at certain abattoirs in Republic of Serbia

Examinations were carried out during the period from January 2006 until March 2007 on a total of 449 samples of the cecum of broilers and the cecum and the colon of pigs. These samples included 251 samples originating from broilers and 198 samples of pig cecums and colons. All the listed samples were obtained by scraping the surface of these parts of the digestive system of broilers and pigs. At the site of sampling, the diluted material was sown on a medium (Karmali agar), in order to get individual colonies. After sowing, the bases were placed in anaerobic jars in which microaerophilic conditions were achieved using Campy Pak, BBL bags. On arrival at the laboratory, the jars containing the sown bases were placed in a thermostat, at a temperature of 42oC for 48 hours for the purpose of incubation. Following incubation, the grown colonies were examined macroscopically, and then microscopic preparations were made from them, which were stained with 2% carbol fuchsin after drying and fixation. Those isolates which were in the form of a comma, the letter S, or gull's wings in the microscopic preparations were considered Campylobacter species (Figures 1 and 2). The isolated thermophilic campylobacteria were identified using conventional and commercial biochemical tests API Campy, manufactured by Bio M?rieux, France. With the application of these microbiological methods, thermophilic Campylobacter species were isolated from 203 (80.88%) of the 251 samples of broiler cecums. Furthermore, thermophilic campylobacteria were isolated from 153 (77.27%) of the 198 samples from the cecum and colon of pigs taken within these investigations. The obtained results indicate that there is a somewhat greater prevalence of these bacteria among the broilers. However, such a high percentage of both broilers and pigs colonized by thermophilic Campylobacter species could pose a serious problem, in particular when it is known that infections of humans caused by the consumption of insufficiently thermally processed meat of broilers or pigs are not infrequent.

Evaluation of a Cytolethal Distending Toxin (cdt) Gene‐Based Species‐Specific Multiplex PCR Assay for the Identification of Campylobacter Strains Isolated from Poultry in Thailand

We have recently developed a cytolethal distending toxin (cdt) gene-based species-specific multiplex PCR assay for identifying Campylobacter jejuni, C. coli and C. fetus. In the present study, the applicability of this assay was evaluated with 34 Campylobacter-like organisms isolated from poultry in Thailand for species identification and was compared with other assays including API Campy, 16S rRNA gene sequence, and hippuricase (hipO) gene detection. Of the 34 strains analyzed, 20, 10 and 1 were identified as C. jejuni, C. coli, and Arcobacter cryaerophilus, respectively, and 3 could not be identified by API Campy. However, 16S rRNA gene analysis, showed that all 34 strains are C. jejuni/coli. To discriminate between these 2 species, the hipO gene, which is specifically present in C. jejuni, was examined by PCR and was detected in 20 strains, which were identified as C. jejuni by API Campy but not in the remaining 14 strains. Collective results indicated that 20 strains were C. jejuni whereas the 14 strains were C. coli. When the cdt gene-based multiplex PCR was employed, however, 19, 20 and 19 strains were identified as C. jejuni while 13, 14 and 13 were identified as C. coli by the cdtA, cdtB and cdtC gene-based multiplex PCR, respectively. Pulsed-field gel electrophoresis revealed that C. jejuni and C. coli strains analyzed are genetically diverse. Taken together, these data suggest that the cdt gene-based multiplex PCR, particularly cdtB gene-based multiplex PCR, is a simple, rapid and reliable method for identifying the species of Campylobacter strains.

血液培養からの<I>Helicobacter cinaedi</I>及びその類縁菌の分離培養と簡便同定法に関する検討

A spiral Gram-negative bacterium, Helicobacter cinaedi, has been isolated from clinical specimens of immunodeficient patients in Japan. We determined the isolation procedure and features using practical phenotypic tests for H. cinaedi isolated from blood culture. We applied 28 clinically isolated strains from 5 different metropolitan institutions, whose species was reconfirmed in technical support at another expert veterinary medicine institute in identifying H. cinaedi. We also used 6 strains of type and reference. Positive signals from aerobic bottles in automated blood culture systems, BACTEC, were observed 4-9 days after culture was started. A thin spread colony was found 3-4 days after the start of subcultures consisting of growth on 5% sheep blood agar plates with immediate incubation under wet and microaerophilic conditions at 35 degrees C. In phenotypic tests, all specimens of H. cinaedi were shown to be positive in nitrate reduction and to be negative in both indoxyl acetate hydrolysis and urease activity, underlining the usefulness of these 3 phenotypes. Tests of alkaline phosphatase hydrolysis, growth at different temperatures, and growth with 1% glycin provided additional information when referencing standard results of identification. We took speculative profiling data of H. cinaedi identification with a simple phenotypic kit, Api campy, indicating its usefulness in testing. These observations suggest combined methods of previously available tests (nitrate reduction, indoxyl acetate hydrolysis, and urease activity) with Api campy may be useful in differentiating H. cinaedi from H. cinaedi-like microorganisms when observing the characteristic thin spread colony within subcultures under microaerophilic conditions.

Presence of Campylobacter spp. in nature

Presence of bacteria from Campylobacter spp. in nature and their importance encourages many researchers to study their biology. The aim of this study was to understand the epizootiology of these bacteria by analyzing all available data and our own results. This would help to follow their movements and prognosticate the outbreak of the disease caused by Campylobacter. In this experiment we used parts of the digestive tracts and reproductive organs of poultry and intestines of pigs and cattle. Isolation methodology was adjusted to the conditions favorable for Campylobacter spp. by providing selective mediums and microaerophilic conditions. Final determination of the isolates was done with Api Campy strips and appropriate software. Of the 60 samples analyzed, 42 were positive for the presence of Campylobacter spp. Campylobacter jejuni in 28 and Campylobacter coli in 14 cases. After examining samples from the reproductive tract, 4 birds were positive, with Campylobacter jejuni in 3 and Campylobacter coli in 1 sample. Of the 12 samples from pigs, 7 were positive (Campylobacter jejuni). Of the 6 samples from calves, Campylobacter jejuni was present in 4 cases. For the sake of clarity, the overall epizootiological situation, established on the basis of literature data and the obtained results, may be viewed as consisting of three interactive parts. The first one is the territorial prevalence of Campylobacter spp. The second is their presence in different kinds of animal and their systems (organs), and the third are health problems that occur in consequence to their presence.

The incidence of Campylobacter spp. on processed turkey from processing plants in the midwestern United States.

The primary aim of this study was to determine the incidence of Campylobacter spp. on turkey, presented for processing at participating production plants located in the midwest region of the United States. The two participating plants were visited on a monthly basis for a period of 1 year. Sampling of carcasses was carried out using a surface swab technique. Swabs were obtained from carcasses at two points on the production line - prechill and postchill. In addition, samples of chill water were also obtained for examination. Isolation and detection of Campylobacter was carried out using enrichment in Preston broth with recovery of the organism on blood free Campylobacter selective agar (CCDA). Isolates recovered were screened and identified using the API Campy identification system. The study found that 34.9% of all samples tested were positive for Campylobacter spp. The overall, contamination rates observed for both plants were relatively similar (39.2% for plant A and 30.6% for plant B). Differences were observed in the incidence of Campylobacter spp. on prechill vs postchill carcasses (i.e. 40.8% prechill vs 37.6% postchill for plant A and 41.8% prechill vs 19.8% postchill for plant B). Campylobacter species most often isolated included Camp. jejuni and Camp. coli. Other species recovered were Camp. fetus fetus, Camp. upsaliensis and Camp. lari. The incidence of Campylobacter spp. on processed poultry was relatively common. Factors such as the processing plant examined, season and the farms presenting birds for processing influenced the incidence of the pathogen. Differences were observed in the prevalence of Campylobacter spp. isolated from the two plants examined. The study suggests a seasonal prevalence of Campylobacter in the cooler months with processing conditions also influencing the overall occurrence of the organism. The incidence, isolation and detection of Campylobacter spp. from processed poultry are discussed.