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Prevalence of thermophilic Campylobacter and Arcobacter butzleri in food of animal origin.

Thermophilic campylobacters and Arcobacter butzleri were looked for in 57 manually shelled egg samples and 130 raw meat samples. No bacteria were found in the egg samples, but they were, however, found in 16.1% of meat samples with a frequency varying from 2.4% (port sausage) to 37.5% (poultry). A total of 21 strains were identified, one belonging to Arcobacter butzleri (pork) and 20 to Campylobacter. The most common species was C. jejuni (16/20) followed by C. coli (4/20) and in both species biotype I was the most frequently found. Regarding the methods employed for the isolation of bacteria, no substancial difference was seen between the three media (Butzler, CCDA and Preston) although CCDA medium gave better results than Butzler and Preston media. The variable which had the most influence was the incubation temperature. A higher number of strains was obtained at 42 degrees C than at 37 degrees C.

Bacteria↗

Isolation of Arcobacter butzleri from ground water.

Arcobacter butzleri was isolated from a contaminated ground water source. These organisms, previously designated as aerotolerant Campylobacter, were capable of surviving in the ground water environment. Specific DNA probes were used to characterize the isolates in the initial identification and survival studies. Arcobacter butzleri was found to be sensitive to chlorine inactivation.

Campylobacter↗

Growth and mechanism of filamentous-sulfur formation by Candidatus Arcobacter sulfidicus in opposing oxygen-sulfide gradients.

Studies were conducted in opposing gradients of oxygen and sulfide in microslide capillaries to (i) characterize the chemical microenvironment preferred by Candidatus Arcobacter sulfidicus, a highly motile, sulfur-oxidizing bacterium that produces sulfur in filamentous form, and (ii) to develop a model describing the mechanism of filamentous-sulfur formation. The highly motile microorganisms are microaerophilic, with swarms effectively aggregating within oxic-anoxic interfaces by exhibiting a chemotactic response. The position of the band was found to be largely independent of the sulfide concentration as it always formed at the oxic-anoxic interface. Flux calculations based on steady state gradients of oxygen and sulfide indicate that sulfide is incompletely oxidized to sulfur, in line with the formation of filamentous sulfur by these organisms. It is proposed that Candidatus Arcobacter sulfidicus effectively competes with other sulfur-oxidizing bacteria in the environment by being able to tolerate higher concentrations of hydrogen sulfide (1-2 mM) and by possessing the ability to grow at very low oxygen concentrations (1-10 muM). The formation of mat-like structures from filamentous sulfur appears to be a population mediated effort allowing these organisms to effectively colonize environments characterized by high sulfide, low oxygen and dynamic fluid movement.

Ecosystem↗

Immunochemical characterization of a haemagglutinating antigen of Arcobacter spp.

The Arcobacter haemagglutinin has been identified by Western immunoblot to be an immunogenic protein of about 20 kDa. The haemagglutinating activity is sensitive to proteolytic enzyme digestion and heat treatment of 80 degrees C and above. The Arcobacter haemagglutinin is possibly a lectin-like molecule binding to erythrocytes via a glycan receptor containing D-galactose as part of its structure.

Animals↗

Outbreak of recurrent abdominal cramps associated with Arcobacter butzleri in an Italian school.

In the autumn of 1983, an outbreak of recurrent abdominal cramps occurred in a nursery and primary school in the Rovigo area in Italy. None of the 10 affected children had diarrhea. An atypical Campylobacter-like organism was isolated from feces in all cases. Conventional enteropathogens were searched for but not detected. The Campylobacter-like organism was identified as Arcobacter butzleri by using sodium dodecyl sulfate-polyacrylamide gel electrophoresis of whole-cell proteins and cellular fatty acid analysis. Its identity was confirmed by DNA-DNA hybridizations versus Arcobacter reference strains. All of the preserved outbreak strains have identical protein profiles and phenotypic characteristics and belong to serogroup 1 of the Lior serotyping scheme on the basis of slide agglutination of crude and absorbed antisera of A. butzleri reference strains versus heat-labile antigens of live bacteria. These data point to an epidemiological relationship. The successive timing of the cases suggests person-to-person transmission.

Campylobacter↗

Occurrence, removal and seasonal variation of thermophilic campylobacters and Arcobacter in sewage sludge.

The presence of thermophilic campylobacters and Arcobacter was investigated in four types of sewage sludge taken from the treatment plant in Bologna (Italy): primary, activated, thickened and anaerobically digested sludge. Campylobacter jejuni and Campylobacter coli were more numerous during the March-September period and were found only in primary sludge (22.7%) with mean counts of 278 MPN/g dry matter and 1403 MPN/g dry matter respectively. Arcobacter butzleri were found in all types of sludge with frequencies of 80% in activated and thickened sludges and 41% in digested sludges. They were more numerous in the spring/summer period with peaks in April, May, June and September. They were less sensitive to anerobic digestion than fecal bacteria, probably due to their microaerophilic growth properties. However, since they are found in anaerobically digested sludges at mean values of 7649 MPN/g dry matter the land application of digested sludges may cause high risks infection.

Campylobacter coli↗

Prevalence of Campylobacter species, Helicobacter pylori and Arcobacter species in stool samples from the Venda region, Limpopo, South Africa: studies using molecular diagnostic methods.

OBJECTIVES: This study determined the prevalence of Campylobacter spp., Helicobacter pylori and Arcobacter spp. in stool samples from Venda in relation to diarrhea, intestinal inflammation and HIV status using specific molecular methods. METHODS: Stool samples were collected from hospital patients (255) and primary school children (67). Genomic DNA was extracted from the stools and molecular methods including PCR, PCR followed by restriction analysis and multiplex PCR were used to test for the different organisms. The lactoferrin content of the stools was determined using commercial kits from TechLab (Blacksburg, VA, USA). RESULTS: The prevalence of the different organisms was 50.6% for H. pylori, 10.2% for C. jejuni, 6.2% for A. butzleri, 6.5% for C. coli, 3.1% for C. concisus, 2.8% for A. cryaerophilus and 1.9% for A. skirrowii. Of all the organisms, only C. jejuni was significantly associated with diarrhea (84.8%) (chi2=21.025, P<0.001) and elevated levels of lactoferrin (78.8%) (chi2=16.919, P<0.005) and was an important pathogen associated with diarrhea among HIV positive individuals (22.8%). CONCLUSIONS: Campylobacter infections are common causes of gastroenteritis in Venda. Non-C. jejuni/coli Campylobacters such as C. concisus as well as A. butzleri and H. pylori may be involved in gastrointestinal diseases in the region but further studies are needed to confirm this hypothesis.

Adolescent↗

Identification of Arcobacter cryaerophilus isolated from a traffic accident victim with bacteremia by 16S ribosomal RNA gene sequencing.

Traditional ways of identifying slow growing bacteria is slow and often difficult. In this study, a small, Gram-negative, facultative anaerobic, slow growing bacillus was isolated from the blood culture of a 7-year old traffic accident victim. The bacterium was non-hemolytic, catalase and oxidase positive. An attempt to use the Vitek system (GNI+) and the API system (20NE) to identify the strain was unsuccessful as the growth controls showed negative results. 16S ribosomal RNA gene sequencing showed that there was 1 base difference between the isolate and Arcobacter cryaerophilus (GenBank Accession no. U25805), 1 base difference between the isolate and A. cryaerophilus (GenBank Accession no. U34387), 10 base differences between the isolate and A. cryaerophilus (GenBank Accession no. L14624), 34 base differences between the isolate and A. butzleri (GenBank Accession no. U34386), 34 base differences between the isolate and A. butzleri (GenBank Accession no. U34387), and 38 base differences between the isolate and A. butzleri (GenBank Accession no. L14626), indicating that the isolate most closely resembled a strain of A. cryaerophilus. Identification of the isolate in our case by conventional methods was difficult, as the absence of a curved morphology has made it confused with other Gram-negative non-fermentative bacteria, and the slow growth rate has made it unidentifiable by both the Vitek and API systems. Although the exact source of infection and route of transmission in our case remains elusive, we speculate that the bacteria were transmitted through the respiratory tract while the boy was suffocated in the mud. The present report represents an example of showing the usefulness of 16S rRNA gene sequencing for identification of slow growing bacteria.

Accidents, Traffic↗

The effect of citric acid, lactic acid, sodium citrate and sodium lactate, alone and in combination with nisin, on the growth of Arcobacter butzleri.

The importance of Arcobacter spp. as a cause of human foodborne illness is unresolved. Organic acids and their sodium salts, and nisin are preservatives commonly used in the type of foods from which the organism is recovered. In this study their effect on the growth of A. butzleri in culture, alone and in combination, was investigated. At 0.5%, 1.0% and 2.0% lactic and citric acids inhibited A. butzleri growth; 2% sodium lactate was effective in inhibiting growth over 8 h incubation but not over longer periods. Sodium citrate was more effective than sodium lactate. Nisin alone inhibited A. butzleri growth at 500 IU ml-1 over 5 h. It did not enhance the effect of sodium citrate inhibition but it did augment the effect of sodium lactate alone over 8 h.

Arcobacter↗

Susceptibility of Arcobacter butzleri isolates to 23 antimicrobial agents.

AIMS: The objective of this study was to determine the susceptibility of Arcobacter butzleri isolates to various antimicrobial agents used in the treatment of infectious diseases in humans and animals. METHODS AND RESULTS: Thirty-nine A. butzleri strains isolated from broiler chickens were tested for their susceptibility to 23 antimicrobial agents using a disc diffusion method. All isolates were resistant to aztreonam, cefuroxime sodium, cephalothin, orbenin, oxacillin, penicillin G and trimethoprim/sulphamethoxazol. Of the 39 isolates tested, 26 were also found resistant to amoxycillin, amoxycillin/clavulanic acid and ampicillin. One isolate was resistant to, and four showed intermediate level of resistance to, erythromycin. All isolates were susceptible to amikacin, chloramphenicol, danofloxacin, enrofloxacin, nitrofurantoin, nalidixic acid, tetracyclines and tobramycin. CONCLUSIONS: The majority of the isolates were found resistant to antibiotics commonly used for the treatment of infectious bacterial diseases in humans and animals. SIGNIFICANCE AND IMPACT OF THE STUDY: This study shows that A. butzleri strains vary in their resistance to certain kinds of antibiotics and caution should be taken when choosing a suitable antibiotic for the treatment of disease(s) caused by this organism.

Animals↗

Discrimination of Arcobacter butzleri isolates by polymerase chain reaction-mediated DNA fingerprinting.

AIMS: The objective of this study was to subtype Arcobacter butzleri isolates using RAPD-PCR. METHODS AND RESULTS: Thirty-five A. butzleri isolates obtained from chicken carcasses were examined. PCR-mediated DNA fingerprinting technique with primers of the variable sequence motifs was used to detect polymorphism within the isolates. Eleven distinct DNA profiles were obtained as follows: Of the 35 strains, 10 as profile 4; seven as profile 1; five as profile 3; three as profiles 2 and 9; two as profile 10; one as profiles 5, 6, 7, 8 and 11. CONCLUSIONS: Chicken carcasses sold in markets were found to be contaminated with several different strains of A. butzleri. RAPD-PCR technique was found to be a useful technique for distinguishing A. butzleri isolates. SIGNIFICANCE AND IMPACT OF THE STUDY: The presence of several different A. butzleri strains on chicken carcasses may indicate multiple sources of contamination. The epidemiological role of A. butzleri in human and animal diseases should be investigated further.

Animals↗

Comparison of sampling techniques for detection of Arcobacter butzleri from chickens.

Arcobacter butzleri is a causative agent of human enteritis that has been recently differentiated from the genus Campylobacter. Previous work suggests that its transmission to humans is likely through a foodborne route with a substantial tendency to be located on poultry carcasses. For reducing the incidence of this pathogen on commercial poultry, improved protocols are needed to sample and identify A. butzleri from infected birds prior to slaughter. The purpose of this study was to compare sampling methods for this emerging pathogen from chickens that were artificially inoculated per os with A. butzleri. We tested three sampling techniques commonly used to determine the microbiological quality of poultry: cloacal swabs, fecal samples, and environmental surface (drag) swabs collected when birds were 3, 5, or 7 wk old. These samples were cultured in Johnson-Murano enrichment broth and analyzed by PCR. Results indicate that environmental surface swabs yielded the highest recovery percentage. A detection rate of 75 to 100% was observed for each sampling period (age of chicken). Additionally, A. butzleri could not be isolated from the intestinal tract (jejunum, ileum, cecum, colorectum) of inoculated birds.

Animals↗

Identification of Campylobacter spp. and discrimination from Helicobacter and Arcobacter spp. by direct sequencing of PCR-amplified cpn60 sequences and comparison to cpnDB, a chaperonin reference sequence database.

A robust method for the identification of Campylobacter spp. based on direct sequencing of PCR-amplified partial cpn60 sequences and comparison of these to a reference database of cpn60 sequences is reported. A total of 53 Campylobacter isolates, representing 15 species, were identified and distinguished from phenotypically similar Helicobacter and Arcobacter strains. Pairwise cpn60 sequence identities between Campylobacter spp. ranged from 71 to 92 %, with most between 71 and 79 %, making discrimination of these species obvious. The method described overcomes limitations of existing PCR-based methods, which require time-consuming and complex post-amplification steps such as the cloning of amplification products. The results of this study demonstrate the potential for use of the reference chaperonin sequence database, cpnDB, as a tool for identification of bacterial isolates based on cpn60 sequences amplified with universal primers.

Arcobacter↗

Genotyping and genetic diversity of Arcobacter butzleri by amplified fragment length polymorphism (AFLP) analysis.

AIMS: To investigate the potential of amplified fragment length polymorphism (AFLP) profiling for genotyping Arcobacter butzleri and to obtain further data on the genetic diversity of this organism. METHODS AND RESULTS: Seventy-three isolates of Danish, British, Turkish, Swedish, Nigerian and North American origin from human infections, chickens, turkeys, ducks, sheep and poultry abbatoir effluent were studied by use of a protocol that involved stringent PCR amplification of fragments derived from digestion of genomic DNA with restriction enzymes BglII and Csp6I. The mean similarity value of duplicate profiles of 10 isolates was 91.15%, indicating the method to be reproducible. Numerical analysis of all 73 isolates distinguished 51 subtypes at the 91% similarity level, of which 39 comprised single strains. The remaining 34 isolates were distributed among 12 subtypes, each of which contained strains homogeneous with respect to their respective source of isolation. However, contemporaneous strains from the same source could also be distinguished. CONCLUSIONS: AFLP profiling is an effective method for typing the genetically diverse organism A. butzleri. SIGNIFICANCE AND IMPACT OF THE STUDY: The study represents a comprehensive analysis of the genetic diversity of A. butzleri by use of isolates from six countries spanning three continents and also shows that several distinct A. butzleri genotypes may be found in a given environment. AFLP profiling appears to have considerable potential for molecular epidemiological studies of this ubiquitous emerging pathogen that is implicated as a causative agent of both human and animal disease.

Abattoirs↗

Genome mapping of Arcobacter butzleri.

A genome map of the potential food-poisoning pathogen Arcobacter butzleri, strain NCTC 12481, has been constructed. The map was ordered by a combination of one- and two-dimensional pulsed field gel electrophoresis, using the restriction enzymes EagI, SacII, SalI and XhoI, and Southern hybridization with digoxygenin-labelled probes. The size of the genome was estimated at 2.57+/-0.01 Mb; this is 0.9 Mb larger than the genome of the closely related species Campylobacter jejuni. We deduce that there are five copies of the 23S and 16S rRNA genes present in the A. butzleri NCTC 12481 genome and two copies of the gene glyA. A gene homologous to pglF, involved in bacterial glycosylation in C. jejuni, was also located on the genome map. In C. jejuni and Wolinella succinogenes, most glycosylation genes are clustered together and the gene order is similar. In A. butzleri NCTC 12481, no analogous organization was found in the region sequenced.

Arcobacter↗

Specific detection of Arcobacter and Campylobacter strains in water and sewage by PCR and fluorescent in situ hybridization.

The aim of this study was to evaluate PCR and fluorescent in situ hybridization (FISH) techniques for detecting Arcobacter and Campylobacter strains in river water and wastewater samples. Both 16S and 23S rRNA sequence data were used to design specific primers and oligonucleotide probes for PCR and FISH analyses, respectively. In order to assess the suitability of the methods, the assays were performed on naturally and artificially contaminated samples and compared with the isolation of cells on selective media. The detection range of PCR and FISH assays varied between 1 cell/ml (after enrichment) to 10(3) cells/ml (without enrichment). According to our results, both rRNA-based techniques have the potential to be used as quick and sensitive methods for detection of campylobacters in environmental samples.

Arcobacter↗

Rapid identification of Campylobacter, Arcobacter, and Helicobacter isolates by PCR-restriction fragment length polymorphism analysis of the 16S rRNA gene.

A rapid two-step identification scheme based on PCR-restriction fragment length polymorphism (PCR-RFLP) analysis of the 16S rRNA gene was developed in order to differentiate isolates belonging to the Campylobacter, Arcobacter, and Helicobacter genera. For 158 isolates (26 reference cultures and 132 clinical isolates), specific RFLP patterns were obtained and species were successfully identified by this assay.

Arcobacter↗

Arcobacter butzleri isolated from a diarrhoeic non-human primate.

The bacteriological examination of a faecal specimen from a 20-year-old female rhesus macaque (Macaca mulatta) with diarrhoeal illness revealed the presence of a large number of a relatively new enteric pathogen, Arcobacter butzleri. The animal was from a closed colony of about 60 females, some of them were showing intermittent diarrhoea possibly related to Giardia spp. Conditions for the isolation and identification of A. butzleri are reported, as well as discussions about its role as a primary pathogen and its zoonotic potential.

Animals↗