Comparative study using amplified fragment length polymorphism fingerprinting, PCR genotyping, and phenotyping to differentiate Campylobacter fetus strains isolated from animals.
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A collection of Campylobacter fetus strains, including both C. fetus subsp. fetus and C. fetus subsp. venerealis, were phenotypically identified to the subspecies level and genotypically typed by PCR and amplified fragment length polymorphism (AFLP) analysis. Phenotypic subspecies determination methods were unreliable. Genotyping of the strains by PCR and AFLP showed a clear discrimination between the two subspecies. (+info)
Campylobacter fetus uses multiple loci for DNA inversion within the 5' conserved regions of sap homologs.
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Campylobacter fetus cells possess multiple promoterless sap homologs, each capable of expressing a surface layer protein (SLP) by utilizing a unique promoter present on a 6.2-kb invertible element. Each sap homolog includes a 626-bp 5' conserved region (FCR) with 74 bp upstream and 552 bp within the open reading frame. After DNA inversion, the splice is seamless because the FCRs are identical. In mutant strain 23D:ACA2K101, in which sapA and sapA2 flanking the invertible element in opposite orientations were disrupted by promoterless chloramphenicol resistance (Cm(r)) and kanamycin resistance (Km(r)) cassettes, respectively, the frequency of DNA inversion is 100-fold lower than that of wild-type strain 23D. To define the roles of a 15-bp inverted repeat (IR) and a Chi-like site (CLS) in the FCR, we mutagenized each upstream of sapA2 in 23D:ACA2K101 by introducing NotI and KpnI sites to create strains 23D:ACA2K101N and 23D:ACA2K101K, respectively. Alternatively selecting colonies for Cm(r) or Km(r) showed that mutagenizing the IR or CLS had no apparent effect on the frequency of the DNA inversion. However, mapping the unique NotI or KpnI site in relation to the Cm(r) or Km(r) cassette in the cells that changed phenotype showed that splices occurred both upstream and downstream of the mutated sites. PCR and sequence analyses also showed that the splice could occur in the 425-bp portion of the FCR downstream of the cassettes. In total, these data indicate that C. fetus can use multiple sites within the FCR for its sap-related DNA inversion. (+info)
Identification, characterization, and variation in expression of two serologically distinct O-antigen epitopes in lipopolysaccharides of Campylobacter fetus serotype A strains.
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Monoclonal antibodies (MAbs) to the lipopolysaccharide (LPS) O-antigens of Campylobacter fetus serotype A and B strains were produced. Eight MAbs specific for serotype A LPS were characterized on immunoblots of C. fetus serotype A LPS. Two immunoblot patterns were observed and were used to divide the eight MAbs into two groups. MAbs M1177 and M1194 were selected as representative of the two groups and were used in an enzyme-linked immunosorbent assay (ELISA) to examine the LPS O-antigen epitopes of 37 serotype A C. fetus subsp. fetus and C. fetus subsp. venerealis strains. Thirty-three strains (89%) reacted with both M1177 and M1194, 2 strains reacted only with M1177, and 2 strains reacted only with M1194. To further characterize the O-antigen epitopes, purified serotype A LPS was treated using various temperature and pH conditions and the effect of the treatments on the reactivity of the LPS with MAbs M1177 and M1194 was evaluated by ELISA. While no difference among several treatments was observed, heating serotype A LPS under alkaline conditions decreased the reaction with M1177 to background levels and increased the reaction with M1194. MAbs M1177 and M1194 were also used with ELISA to investigate in vivo and in vitro expression of the two O-antigen epitopes. There was substantial variation in expression of the two epitopes among weekly isolates of two C. fetus serotype A strains recovered from experimentally infected heifers. There was minimal variation in expression of the two epitopes in successive subcultures of three C. fetus serotype A strains. (+info)
Campylobacter-induced fetal death in a rhesus monkey.
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A pregnant 4-year-old rhesus monkey (Macaca mulatta) was presented with a history of acute vaginal bleeding. Physical examination revealed an open cervix. An ultrasound scan demonstrated a viable early third-trimester fetus, approximately 16 weeks of gestational age. Hematology results showed that the monkey was anemic, with a normal leukogram and Dohle bodies. A subsequent cervical culture was positive for Campylobacter fetus. The fetus died 3 days later, and a necropsy of the fetus and placenta was performed. Microscopic examination of the placenta revealed villitis, perivillitis, and deciduitis with S-shaped and gull wing-shaped bacteria. C. fetus was considered the cause of the placental lesions and fetal death; however, the pathogenesis of the infection (hematogenous vs. ascending from the maternal genital tract) was not clear. This is the first report of a Campylobacter-induced fetal death in the rhesus monkey. Because macaques can be asymptomatic carriers and Campylobacter-induced diarrhea is common, this finding has implications for breeding success in nonhuman primate breeding colonies. (+info)
Thermo-tolerant Campylobacter fetus bacteraemia identified by 16S ribosomal RNA gene sequencing: an emerging pathogen in immunocompromised patients.
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Eight Campylobacter isolates that were able to grow at 25 degrees C and 42 degrees C and had the same biochemical profile were isolated from the blood of eight immunocompromised patients. Conventional biochemical tests were unable to determine whether they were isolates of thermo-tolerant C. fetus, H2S-negative C hyointestinalis, or a new Campylobacter species. Sequencing of the 16S ribosomal RNA genes showed that all eight isolates had the same nucleotide sequence and this was identical to that of C. fetus (GenBank accession no. AF219233). All eight patients had underlying disease and two died despite antibiotic treatment. Because of the ability of C fetus to grow over a wide range of temperatures and a higher incidence of bacteraemia by this organism than C. jejuni in the past 5 years in Hong Kong, thermo-tolerant C fetus may be an emerging pathogen in immunocompromised patients in the years to come. (+info)
Isolation of Campylobacter fetus subsp. fetus from a patient with cellulitis.
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Campylobacter fetus subsp. fetus is a gram-negative, slender, spirally curved bacterial pathogen. It has been isolated from human blood, spinal fluid, and abscesses, but cellulitis associated with bacteremia is rare. We report its isolation from a blood culture of a human patient with cellulitis as well as difficulties encountered in determining the identity of the subspecies of C. fetus. (+info)
Role of S-layer protein antigenic diversity in the immune responses of sheep experimentally challenged with Campylobacter fetus subsp. fetus.
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Surface layer proteins (SLPs) are essential for induction of abortion by Campylobacter fetus subsp. fetus in experimentally challenged ewes. These proteins are encoded by multiple sap genes and vary in size and antigenicity. The role of SLP antigenic variation during experimental ovine infection was investigated. Following subcutaneous challenge, the SLPs were highly antigenic, and antibodies were detected in serum, milk, bile, and urine. Fecal anti-SLP antibodies were detected only in animals challenged orally. Ewes challenged with wild-type strain 23D with variable SLPs developed detectable circulating anti-SLP immunoglobulin G (IgG) antibodies by 2 weeks postchallenge. In contrast, ewes challenged with mutants of 23D that had fixed expression of a single SLP developed antibodies within 1 week postchallenge, suggesting that antigenic variation in SLPs may delay the host antibody response. Although not statistically significant, the data from challenge experiments in which vaccinated ewes were used suggested that SLP-expressing vaccines could protect animals from abortion and that this effect was independent of the SLP expressed, indicating involvement of conserved epitopes in the SLP. The conserved 184-amino-acid N-terminal region of the SLP, identified from previously published sequences, was epitope mapped with rabbit anti-SLP antisera by using overlapping synthetic 20-mer peptides. Two putative epitopes were identified at amino acids 81 to 110 and 141 to 160. Amino acids 81 to 100 also bound serum IgG antibodies from experimentally challenged sheep. Conserved antigenic regions of the SLP that induce protective immune responses may enable development of synthetic vaccine candidates for C. fetus subsp. fetus-associated ovine abortion. (+info)
Epidemiology and antimicrobial susceptibilities of 111 Campylobacter fetus subsp. fetus strains isolated in Quebec, Canada, from 1983 to 2000.
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The epidemiology and antimicrobial susceptibilities of 111 Campylobacter fetus subsp. fetus strains isolated from 103 patients from 1983 to 2000 in Quebec, Canada, were determined. The median number of patients infected annually with this bacteria was seven, with an incidence of 0.1 per 100,000 population. The male-to-female ratio was 1.1 to 1.0. The patients originated from 13 of the 18 Quebec socioeconomic regions. The age range of the patients was 6 months to 90 years old, 53% being > or = 70 years old and 2% being <20 years old. The isolation site was blood for 69% of the patients, stools for 20%, and other body fluids for 11% of them. Three patients suffered a relapse, with the same strain being isolated from the same site at different times as confirmed by pulse-field gel electrophoresis. All isolates were susceptible to ampicillin, gentamicin, meropenem, and imipenem, with 90% minimal inhibitory concentrations of 4, 1, 0.12, and < or = 0.06 microg/ml, respectively. Three percent and two percent of the strains were, respectively, resistant and intermediate to ciprofloxacin. Thirty-four percent of the strains were resistant to tetracycline. There was a nonsignificant increase in resistance to ciprofloxacin (P = 0.27) and to tetracycline (P = 0.65) in recent years. The percentages of intermediate and resistant MICs were, respectively, 12 and 1% for cefotaxime and 71 and 0% for erythromycin. All strains were beta-lactamase negative. (+info)