Lytic Activity of Various Phage Cocktails on Multidrug-Resistant Bacteria.

PURPOSE: A bacteriophage is a virus that infects and replicates within a bacterium following the injection of phage genome into the bacterial cytoplasm. They are seen as a possible therapy for multi-drug-resistant strains of many bacteria. The aim of this study is to evaluate the lytic activity of the Pyo, Intesti and Fersisi bacteriophage cocktails on P. aeruginosa and S. aureus. METHODS: Ten different S. aureus and P. aeruginosa strains, which were isolated from hospitalized patients in Turkey, were used in the study. The identification and antibiotic susceptibility of the isolates were performed using Vitec 2 system. The identities of the isolates were confirmed by a species-specific Polymerase Chain Reaction (PCR) assay. Lytic activity of the bacteriophage cocktails on bacteria was determined by spot test and plaque assay methods. RESULTS: The lytic activity of the Pyo phage cocktail was evaluated on P. aeruginosa and S. aureus strains. It was found that eight isolates of MDR S. aureus were susceptible to Pyo phage cocktail and two isolates were resistant. Nine isolates of antibiotic-resistant P. aeruginosa were found to be susceptible to this phage cocktail and one isolate was resistant. Thus, the Pyo, Intesti and Fersisi cocktails are very effective in treating clinical strains of multidrug-resistant P. aeruginosa and S. aureus isolated in Turkey. CONCLUSION: The Pyo, Intesti and Fersisi cocktails may prove useful in the treatment of various infections caused by those bacteria.

Emerging pathogen Arcobacter spp. in acute gastroenteritis: molecular identification, antibiotic susceptibilities and genotyping of the isolated arcobacters.

The aims of this study were as follows: (i) to isolate Arcobacter spp. from the stool samples of patients with gastroenteritis; (ii) to identify them with molecular methods; (iii) to genotype them using enterobacterial repetitive intergenic consensus (ERIC)-PCR; and (iv) to determine their antibiotic susceptibilities. For the study, a total of 3287 diarrhoeal stool samples submitted to the Microbiology Laboratory of the Kayseri Training and Research Hospital, Kayseri, Turkey, between 2010 and 2011 were analysed. Campylobacter blood-free selective medium supplemented with cefoperazone, amphotericin B and teicoplanin was used for isolation. Medium inoculated with stool samples was incubated microaerobically at 37 °C for 72-96 h. Phenotypic tests, a genus-specific PCR and a multiplex PCR were used to identify the arcobacters, whilst ERIC-PCR was used for genotyping and the antibiotic susceptibilities of the isolates were detected by E-test. Arcobacter spp. were isolated from nine of the 3287 samples. These nine isolates were identified as Arcobacter butzleri and all showed different ERIC-PCR profiles. All nine isolates were resistant to ampicillin and susceptible to gentamicin, tetracycline, erythromycin and ciprofloxacin. As far as is known, this is the first study in which A. butzleri has been isolated from human acute gastrointestinal infections in Turkey. According to these results, it is recommended that, when investigating the aetiology of infections of the digestive system in humans, Arcobacter spp. be considered for inclusion. The results of this study should contribute to our knowledge related to A. butzleri infections in humans.

Seroepidemiological investigation of brucellosis in sheep abortions in Kars, Turkey.

This study was undertaken to investigate the seroprevalence of brucellosis in unvaccinated sheep from the flocks having previous abortion cases in Kars and around, Turkey and to compare the efficacy of each serological test used. Four hundred serum samples collected from 16 different flocks of sheep having a history of abortions in Kars and its surrounding area in Turkey were examined for the presence of antibodies raised against Brucella using Rose Bengal Plate Test (RBPT), Serum Agglutination Test (SAT), Rivanol Agglutination Test (RAT) and Complement Fixation Test (CFT). All animals were unvaccinated against Brucella. Of the serum samples tested, 147 (%36.7), 142 (%35.5), 139 (%34.75) and 135 (%33.75) were found positive by SAT, RAT, RBPT and CFT, respectively. No statistically significant difference was found between the serological tests used (p > 0.05). It is concluded from this study that brucellosis continues to be an important problem for ovine abortions and poses a risk both for human and other animals in this area. Therefore, adequate intervention measures should be implemented to control and eradicate brucellosis. In addition, if conventional serological tests are used at least two tests, RPBT for screening and CFT for the confirmation of the positive samples are preferable, should be used in parallel for detection of brucellosis effectively.

Isolation of various Arcobacter species from domestic geese (Anser anser).

In this study, the prevalence and distribution of various Arcobacter spp. were investigated in samples taken from the cloacae of healthy domestic geese raised in Turkey. A membrane filtration technique with a non-selective blood agar was employed after enrichment in Arcobacter enrichment broth (AEB) to isolate a wide range of Arcobacter spp. In addition, the isolates were characterized phenotypically and identified at species level using a multiplex-PCR assay. A total of 90 cloacal swab samples taken from geese, collected on three farms (18, 25, 47 samples, respectively), were examined. Of the samples examined, 16 (18%) were found positive for Arcobacter. One Arcobacter species was isolated from each bird. Of the 16 Arcobacter isolates, 7 (44%), 7 (44%) and 2 (12.5%) were identified by m-PCR as A. cryaerophilus, A. skirrowii and A. butzleri, respectively. The present study indicates that domestic geese can harbour a variety of Arcobacter spp. in their cloacae. The presence of Arcobacter in geese may be of significance as reservoirs in their dissemination. Detailed research is needed for better understanding of the epidemiology and zoonotic potential of this emerging pathogen.

Prevalence and distribution of Arcobacter species in various sources in Turkey and molecular analysis of isolated strains by ERIC-PCR.

AIMS: To determine the prevalence of Arcobacter in various food, animal and water sources in Turkey and to subtype the isolated strains using enterobacterial repetitive intergenic consensus (ERIC)-PCR. METHODS AND RESULTS: A total of 806 samples consisting of chicken (100) and turkey meat (100); minced beef (27); rectal swabs from cattle (173), sheep (68) and dogs (62); cloacal swabs of broilers (100) and layers (100); gall bladders of cattle (50) and drinking water samples (26) were examined. A previously described membrane filtration method was used for the isolation. Isolates were identified at species level using multiplex-PCR and discriminated by ERIC-PCR for subtyping. Ninety-eight (12.1%) of the samples examined were found positive for arcobacters. Arcobacter spp. were isolated from 68%, 4%, 6.9%, 8% and 37% of chicken and turkey meats, rectal swabs and gall bladders of cattle and minced beef, respectively. No arcobacters were obtained from the rectal swabs of sheep and dogs, cloacal swabs of broilers and layers, and water samples examined. In total, 99 Arcobacter isolates were obtained. Of these isolates, 92 were identified as Arcobacter butzleri, five were Arcobacter skirrowii and two were Arcobacter cryaerophilus. Thirteen distinct DNA profiles among A. butzleri isolates were obtained by the ERIC-PCR. Of these profiles, eight were from chicken carcass, three from cattle rectal swab and two from minced beef meat isolates. Some of the isolates originated from different sources gave the same DNA profiles. All isolates of A. skirrowii and A. cryaerophilus gave different DNA profiles. CONCLUSIONS: Poultry carcasses, minced beef meat, rectal swabs and gall bladders of cattle were found to be positive for Arcobacter spp. A. butzleri was the predominant species isolated. In addition, large heterogeneity among the Arcobacter isolates was determined. SIGNIFICANCE AND IMPACT OF THE STUDY: Contamination of the poultry carcasses and minced beef meat, rectal and gall bladder samples of cattle with arcobacters poses a risk for both human and animal infections. Detection of several different Arcobacter strains may suggest multiple sources for contamination and infection.

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.

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.

Investigation of arcobacters in meat and faecal samples of clinically healthy cattle in Turkey.

AIMS: To investigate the presence of Arcobacter spp. in minced beef meat (n = 97) and rectal faecal samples (n = 200) collected from cattle immediately after slaughter at a local abattoir in Turkey. METHODS AND RESULTS: Meat samples were examined using three different isolation procedures (CAT-supplemented media, de Boer arcobacter isolation method and membrane filtration method), but only one method (CAT-supplemented media) was employed for faecal samples. The isolated Arcobacter strains were identified by genus- and species-(multiplex) specific PCR assays. Arcobacter spp. were isolated from 5 and 9.5% of meat and faecal samples respectively. Although the only Arcobacter sp. found in meat samples was Arcobacter butzleri, all three pathogenic species--A. butzleri, A. cryaerophilus and A. skirrowii--were detected in the rectal swabs. No Arcobacter was isolated when the de Boer method was used for minced meat samples but the same five meat samples were found positive for arcobacters when CAT-supplemented media and membrane filtration method were used. CONCLUSIONS: The membrane filtration method was found to be superior to the CAT-supplemented media, because it led to a reduction in competing microflora. However, the necessity for one filter and medium for each sample makes this method somewhat expensive. The multiplex-PCR (m-PCR) assay shortened significantly the time required for the identification of Arcobacter spp. and also removed the possibility of false positive results due to other campylobacteria. SIGNIFICANCE AND IMPACT OF THE STUDY: This study reports the isolation of Arcobacter spp. in cattle for the first time in Turkey. The m-PCR assay enables the identification and differentiation of all arcobacters simultaneously in one-step PCR.

Differentiation of Arcobacter species by numerical analysis of AFLP profiles and description of a novel Arcobacter from pig abortions and turkey faeces.

AIMS: To evaluate the efficacy of amplified fragment length polymorphism (AFLP)-based genetic profiling for taxonomic and epidemiological analyses of diverse Arcobacter species. METHODS AND RESULTS: Seventy-two isolates of A. butzleri, A. cryaerophilus, A. skirrowii and A. nitrofigilis, and a previously unclassified porcine abortion strain were studied. AFLP profiling was performed using a BglII-Csp6I-based protocol previously used to characterize Campylobacter species. Duplicate profiles of 20 isolates were 93.25% similar, indicating high reproducibility. Numerical analysis of all 72 strains revealed five phenons at the 29% similarity level, four of which represented each of the known species studied. The remaining phenon was further characterized by phenotypic and 16S rDNA sequence analyses, the results of which indicated it to be a novel Arcobacter species. The genetically distinct subgroups of A. cryaerophilus were differentiated at the 39.5% similarity level. For strain typing, 62 distinct types were defined, with evidence of clonal lineages within A. butzleri, A. cryaerophilus and A. skirrowii. CONCLUSIONS: AFLP profiling is an effective means of determining taxonomic and strain relationships for arcobacters. SIGNIFICANCE AND IMPACT OF THE STUDY: First use of AFLP profiling for diverse Arcobacter species; indication of clonality in A. butzleri, A. cryaerophilus and A. skirrowii; potentially novel Arcobacter taxon identified.

The prevalence of Arcobacter spp. on chicken carcasses sold in retail markets in Turkey, and identification of the isolates using SDS-PAGE.

In this study, the prevalence of Arcobacter spp. on chicken carcasses sold in various retail markets in Turkey was investigated. The isolates were characterized and identified using various phenotypic and molecular tests. The membrane filtration technique employing 0.45-microm pore size membrane filters laid onto a nonselective blood agar was used after enrichment in Oxoid Arcobacter Enrichment Broth (AEB) to examine a total of 75 chicken carcasses (44 fresh and 31 frozen). Species level identification was performed using SDS-PAGE of whole-cell proteins and a recently developed multiplex-PCR assay. All isolates were identified as Arcobacter butzleri. Of the 44 fresh chicken carcasses examined, 42 (95%) were positive for A. butzleri. A. butzleri was also recovered from seven (23%) of the 31 frozen carcasses examined.

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.

The isolation and characterization of Campylobacter jejuni subsp. jejuni from domestic geese (Anser anser).

AIMS: The objectives of this study were to determine the presence of thermophilic Campylobacter spp. in free range domestic geese, and to characterize isolated strains using phenotyping criteria and SDS-PAGE of whole-cell proteins. METHODS AND RESULTS: Forty cloacal swabs from two different flocks of domestic geese were examined. All Camp. jejuni strains isolated from geese were biotyped using the Lior biotyping scheme. Twelve Camp. jejuni isolates were also tested for their susceptibility to 17 different antibacterial agents by a disc diffusion METHOD: Fourteen of the isolates were also subjected to SDS-PAGE. All of the geese examined were found to harbour Camp. jejuni. Six geese carried more than one species of Campylobacter. All strains examined were susceptible to various antibiotics but resistant to penicillin G and cephalothin. Eleven strains (92%) were resistant to sodium cefuroxime, and eight (67%) were resistant to cloxacillin, ampicillin and colistin sulphate. Three strains (25%) were resistant to tetracycline, and one strain was resistant to sulfamethoxazole/trimethoprim and kanamycin. Nine strains were subtyped as Camp. jejuni subsp. jejuni biotype II and the remaining ones as biotype I. There were 96% and 100% similarities between all the strains examined by SDS-PAGE. CONCLUSION: This study showed that Camp. jejuni were common in the intestinal tract of domestic geese. SIGNIFICANCE AND IMPACT OF THE STUDY: Geese should be considered as potential reservoirs for human and animal campylobacteriosis. The antibiotic resistance data from this study also showed that fluoroquinolone resistance, which appears to be a problem in poultry isolates in some countries, is not yet a problem in these geese.

Clonality of Campylobacter sputorum bv. paraureolyticus determined by macrorestriction profiling and biotyping, and evidence for long-term persistent infection in cattle.

Eighteen strains of Campylobacter sputorum bv. paraureolyticus (isolated over a 12-month period from seven dairy cows contained in a single herd) were examined by resistotyping, and macrorestriction profiling using pulsed field gel electrophoresis (PFGE). The resistotypes of these strains were identical, although repeat testing indicated resistance to metronidazole was not a reliable trait for typing purposes. Five SmaI-derived genotypes were identified among the 18 strains. In 5 of 7 cows, isolates obtained from the same animal, but from different time periods, were genotypically indistinguishable, indicating persistence of infection. Macrorestriction profiles of 5 strains representing the 5 SmaI genotypes and 8 other strains of C. sputorum from various sources, were prepared using 4 endonucleases (SmaI, SalI, BamHI and KpnI). The only other strain of C. sputorum bv. paraureolyticus examined (a Canadian isolate from human faeces), was found to have a SmaI macrorestriction profile identical with one of the five clones isolated from the cattle. Moreover, SalI and BamHI profiles of all bv. paraureolyticus strains were similar, while digestion with KpnI was not observed. By contrast, the seven strains of C. sputorum bv. sputorum yielded various macrorestriction profiles with all the enzymes used, and features distinguishing the two biovars studied could be identified. This study indicates that C. sputorum can persist in cattle for at least 12 months and exhibits a clonal population genetic structure.

Diversity and prevalence of Arcobacter spp. in broiler chickens.

Ninety-nine strains of Arcobacter spp., isolated from 10 chicken carcasses purchased from a supermarket and 15 chicken carcasses collected from a poultry abattoir, were speciated using a variety of phenotypic identification methods. All were tested using API Campy test strips and the 16-test (Preston) identification scheme developed for campylobacters. Fifty strains were selected for examination using a more comprehensive probabilistic identification scheme, and the identity of representative strains confirmed by protein profiling using SDS-PAGE. All 25 carcasses yielded Arcobacter butzleri. Three supermarket and 10 abattoir carcasses also carried A. cryaerophilus, and two abattoir carcasses carried A. skirrowii. The API Campy scheme proved unsatisfactory for identifying these strains: only 20 of 99 strains were accurately identified, all of which were A. cryaerophilus, the only Arcobacter sp. included in the database. Moreover, 76 of 99 strains were misidentified. The 16-test scheme identified all the arcobacter strains as A. cryaerophilus, since neither A. butzleri nor A. skirrowii had been described when the scheme was developed. The computer-assisted probabilistic scheme succeeded in identifying all but one strain, the identity of which was clarified by the use of SDS-PAGE. To our knowledge this is the first time that arcobacters other than A. butzleri have been reported in poultry meat or any other food of animal origin. Their high prevalence in poultry products may be of significance to public health.

Identification of unusual Campylobacter-like isolates from poultry products as Helicobacter pullorum.

Twenty-six unclassified Campylobacter-like strains previously isolated from 15 chicken carcasses and caecal contents, together with two more strains isolated from chicken faeces on a different occasion, were identified as Helicobacter pullorum using various phenotypic identification methods. API Campy identification kits and a 16-test identification scheme developed for campylobacters failed to identify these bacteria, or identified them as Campylobacter spp. Eighteen strains (including the two isolated on a different occasion) were chosen for examination using a more comprehensive probabilistic identification scheme. Using this method, 14 of the 18 strains were identified as H. pullorum with ID scores > 95%; two strains were also identified as H. pullorum with lower ID scores. Of the remaining two strains, one was not identified with this scheme and the other was misidentified to the H. acinonyx pylori complex. Whole cell protein profiling by SDS-PAGE confirmed the identity of these isolates as H. pullorum, affirming the value of a polyphasic approach for accurately identifying campylobacteria. The comparatively high prevalence of H. pullorum in poultry determined in this study (60%) suggests that routine isolation and identification methods should be amended to enable a thorough evaluation of its role in human gastroenteritis and avian hepatitis. Some phenotypic characters useful in identifying poultry campylobacteria are presented which could be utilized, along with other technique(s), for improved differentiation of the campylobacteria that are found in poultry.

The isolation and prevalence of campylobacters from dairy cattle using a variety of methods.

Faecal samples from 94 dairy cows and 42 calves in three different herds were examined by a variety of techniques for campylobacters. Cefoperazone amphotericin teicoplanin (CAT) agar, modified cefoperazone charcoal deoxycholate agar (mCCDA), Karmali agar, and membrane filtration onto blood agar, were used with and without enrichment in CAT broth. Seventy-nine percent of cattle in herd A carried campylobacters, compared with 40% and 37.5% of cattle in herds B and C, respectively. Most animals carried only one species of Campylobacter. Campylobacter hyointestinalis was isolated most frequently (32% animals positive) with Camp. fetus subsp. fetus and Camp. jejuni subsp. jejuni detected in 11% and 7% of animals, respectively. In addition, a novel biotype of Camp. sputorum was isolated from 60% of 47 cows tested in herd A. Direct plating detected only two of the total of 40 animals positive for campylobacter. Enrichment in CAT broth before membrane filtration onto blood agar or CAT agar were the most successful methods of plating. Campylobacter sputorum was isolated from CAT agar and blood agar but not from mCCDA or Karmali agar. Karmali agar incubated at 30 degrees C was especially effective for isolating Camp. fetus subsp. fetus.

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.

Emended description of Campylobacter sputorum and revision of its infrasubspecific (biovar) divisions, including C. sputorum biovar paraureolyticus, a urease-producing variant from cattle and humans.

A polyphasic taxonomic study of 15 bovine and human strains assigned to the catalase-negative, urease-positive campylobacter (CNUPC) group identified these bacteria as a novel, ureolytic biovar of Campylobacter sputorum for which we propose the name C. sputorum bv. paraureolyticus: suitable reference strains are LMG 11764 (human isolate) and LMG 17590 (= CCUG 37579, bovine isolate). The present study confirmed previous findings showing that the salient biochemical tests used to differentiate C. sputorum bv. sputorum from C. sputorum bv. bubulus are not reproducible; and that the absolute validity of source-specific biovars of the species is questionable. A correlation between the results of numerical analysis of protein profiles and the reaction of strains in certain enzyme tests was, however, noted. Therefore, it is proposed that the infrasubspecific (biovar) divisions of C. sputorum should be revised to include bv. sputorum for catalase-negative strains; bv. fecalis for catalase-positive strains; and bv. paraureolyticus for urease-positive strains. Strains classified previously as bv. bubulus should be reclassified as bv. sputorum. The species description of C. sputorum is revised accordingly.

Isolation and characterization of a novel catalase-negative, urease-positive Campylobacter from cattle faeces.

Forty-four strains of a phenotypically unique Campylobacter were isolated from the faeces of 26 of 45 cows in a single herd. Isolation involved enrichment and membrane filtration onto blood agar or plating onto cefoperazone amphotericin teicoplanin agar. The strains exhibited phenotypic characteristics typical for Campylobacter species. However, they were unusual in that they produced urease and copious H2S in triple sugar iron (TSI) medium, but did not produce catalase. They did not grow aerobically. None of the strains grew on modified cefoperazone charcoal deoxycholate agar (mCCDA). Macrorestriction profiles of chromosomal DNA were prepared for 15 strains using pulsed-field gel electrophoresis (PFGE). Twelve of 15 profiles were identical and all appeared to be closely related. These catalase-negative, urease-positive campylobacters (CNUPC) represent a group not previously reported. Their sensitivity to antibiotics normally used in selective media for campylobacters might explain why they have not previously been encountered. Their ecological significance and importance with respect to human and animal disease remain to be assessed.