Effect of iron concentration on toxin production in Campylobacter jejuni and Campylobacter coli

1986 ◽  
Vol 32 (5) ◽  
pp. 395-401 ◽  
Author(s):  
Barbara A. McCardell ◽  
Joseph M. Madden ◽  
John T. Stanfield
Keyword(s):  
Campylobacter Jejuni ◽  
Ferrous Sulfate ◽  
Culture Media ◽  
Iron Concentration ◽  
Toxin Production ◽  
Enzyme Linked Immunosorbent Assay ◽  
Campylobacter Coli ◽  
Casamino Acids ◽  
Culture Supernatants ◽  
Campylobacter Spp

The effect of iron concentrations in culture media on supernatant yields of Campylobacter cytotonic toxin (CCT) was studied. Of the 118 Campylobacter spp. strains surveyed, 78.8% produced toxin in brucella broth or in casamino acids – yeast extract (CYE) broth. When the iron concentration of CYE was increased from 0.44 μg/mL (7.9 μM) to 0.65 μg/mL (11.6 μM) by the addition of ferric chloride, 94.9% of the strains were positive for toxin in a ganglioside GM1 based, enzyme-linked immunosorbent assay, using antibody to affinity-purified CCT. The addition of iron as ferrous sulfate was less effective. When four toxin-positive strains were grown in a deferrated medium of conalbumin-treated CYE with 0.04–0.08 μg iron/mL (0.72–1.43 μM), two of the culture supernatants became negative (absorbance at 410 nm, < 0.1 and < 10 ng CCT/mL), and two produced about 90% less CCT but were still classified as positive (absorbance, ≥ 0.1 and ≥ 10 ng CCT/mL). It was therefore concluded that the production of CCT by Campylobacter spp. is influenced by iron concentration.

Prevalence and Antimicrobial Resistance of Campylobacter spp. Isolated from Poultry Carcasses in Poland

2013 ◽  
Vol 76 (8) ◽  
pp. 1451-1455 ◽  
Author(s):  
KINGA WIECZOREK ◽  
IWONA KANIA ◽  
JACEK OSEK
Keyword(s):  
Campylobacter Jejuni ◽  
23S Rrna ◽  
Rrna Gene ◽  
Campylobacter Coli ◽  
Genetic Determinants ◽  
Gyra Gene ◽  
23S Rrna Gene ◽  
Pcr Method ◽  
Almost All ◽  
Campylobacter Spp

The purpose of the present study was to determine the prevalence of Campylobacter in poultry carcasses at slaughter in Poland. For the isolated strains, resistance to selected antibiotics and the associated genetic determinants were identified. A total of 498 Campylobacter isolates were obtained from 802 poultry samples during the 2-year study period. Strains were identified to species with the PCR method; 53.6% of the strains were Campylobacter jejuni and 46.4% were Campylobacter coli. A high percentage of the tested Campylobacter strains were resistant to ciprofloxacin and nalidixic acid (74.1 and 73.5%, respectively) followed by tetracycline (47.4%) and streptomycin (20.5%). Only one C. jejuni and two C. coli isolates were resistant to gentamicin. Seventy-nine (15.9%) of the 498 strains were resistant to three or more classes of antibiotics examined. Higher levels of resistance, irrespective of the antimicrobial agent tested, were found within the C. coli group. Almost all strains resistant to quinolones (99.5%) and to tetracycline (99.6%) carried the Thr-86-to-Ile mutation in the gyrA gene and possessed the tet(O) marker, respectively. All isolates resistant to erythromycin had the A2075G mutation in the 23S rRNA gene. These results reveal that poultry carcasses in Poland are a reservoir of potentially pathogenic and antimicrobial-resistant Campylobacter strains for humans, which may pose a public health risk.

scholarly journals Multiplex PCR Assay for Identifi cation and Differentiation of Campylobacter jejuni and Campylobacter coli Isolates

Folia Medica ◽  
2016 ◽  
Vol 58 (2) ◽  
pp. 95-100 ◽  
Author(s):  
Maria R. Pavlova ◽  
Elina G. Dobreva ◽  
Katucha I. Ivanova ◽  
Galina D. Asseva ◽  
Ivan N. Ivanov ◽  
...  
Keyword(s):  
Multiplex Pcr ◽  
Campylobacter Jejuni ◽  
Accurate Method ◽  
Clinical Isolates ◽  
Campylobacter Coli ◽  
Pcr Assay ◽  
Biochemical Tests ◽  
Multiplex Pcr Assay ◽  
Hydrolysis Of ◽  
Campylobacter Spp

AbstractIntroduction: Campylobacter spp. are important causative agents of gastrointestinal infections in humans. The most frequently isolated strains of this bacterial genus are Campylobacter jejuni and Campylobacter coli. To date, genetic methods for bacterial identification have not been used in Bulgaria. We optimized the multiplex PSR assay to identify Campylobacter spp. and differentiate C. jejuni from C. coli in clinical isolates. We also compared this method with the routinely used biochemical methods.Aim: To identify Campylobacter spp. and discriminate C. coli from C. jejuni in clinical isolates using multiplex PCR assay.Materials and methods: Between February 2014 and January 2015 we studied 93 stool samples taken from patients with diarrheal syndrome and identified 40 species of Campylobacter spp. in them. The clinical material was cultured in microaerophilic atmosphere, the isolated strains being biochemically diff erentiated (hydrolysis of sodium hippurate for C. jejuni, and hydrolysis of indoxyl acetate for C. coli). DNA was isolated from the strains using QiaAmp MiniKit (QIAGEN, Germany). Twenty strains were tested with multiplex PCR for the presence of these genes: cadF, characteristic for Campylobacter spp., hipO for C. jejuni and asp for C. coli.Results and discussion: The biochemical tests identified 16 strains of C. jejuni, 3 strains of C. coli, and 1 strain of C. upsaliensis. After the multiplex PCR assay the capillary gel electrophoresis confirmed 16 strains of C. jejuni, 2 strains of C. coli and 2 strains of Campylobacter spp. - because of the presence of the gene cadF. C. jejuni has the gene hipO, and it is possible that this gene may not be expressed in the biochemical differentiation yielding a negative reaction as a result. In comparison, we can conclude that the genetic differentiation is a more accurate method than the biochemical tests.Conclusion: The multiplex PCR assay is a fast, accurate method for identifi cation of Campylobacter spp. which makes it quite necessary in the clinical diagnostic practice.

Detection of Campylobacter jejuni and Campylobacter coli from Broiler Chicken–Related Samples Using BAX PCR and Conventional International Organization for Standardization Culture

2008 ◽  
Vol 71 (4) ◽  
pp. 835-838 ◽  
Author(s):  
LISA K. WILLIAMS ◽  
ALISDAIR MCMEECHAN ◽  
TAMSIN BAALHAM ◽  
LAURA WARD ◽  
TOM J. HUMPHREY ◽  
...  
Keyword(s):  
Campylobacter Jejuni ◽  
Broiler Chicken ◽  
Culture Method ◽  
International Organization ◽  
Campylobacter Coli ◽  
Poultry Production ◽  
Production Chain ◽  
Enrichment Cultures ◽  
International Organization For Standardization ◽  
Campylobacter Spp

In this study, the conventional International Organization for Standardization (ISO) culture method was compared with the DuPont Qualicon BAX system, a high-throughput, rapid molecular assay that can be used to detect several bacterial species, including Campylobacter jejuni and Campylobacter coli in diverse sample types. Standard enrichment culture is a time-consuming process, taking up to 6 days to obtain a confirmed result. Rapid molecular assays have been developed that provide results within 24 h. Naturally contaminated samples from the poultry production chain were examined for the presence of Campylobacter spp. Samples from broiler chicken ceca (n = 100), fresh chicken carcass rinses (n = 60), and bootsocks (gauze sock walked through a broiler chicken house; n = 50) were enriched according to the ISO 10272 method in Bolton broth specifically designed to detect Campylobacter spp. in complex sample types. Samples were enriched without blood for use with the BAX system using the Campylobacter BAX kits for the detection of C. jejuni and C. coli. Samples also were directly plated onto modified charcoal cefperazone deoxycholate agar, and results were compared with those from the enriched samples for the ability to detect Campylobacter spp. Campylobacter spp. were isolated from 49% of samples with conventional enrichment cultures, from 48% with direct culture, from 68% with the BAX system and enrichment cultures, and from 62% with the BAX system used directly with samples. Overall, the BAX system detected more positive samples than did the conventional culture method and is an effective methodology for the rapid and reliable detection of Campylobacter spp. from diverse sample types.

Prevalence of putative virulence markers inCampylobacter jejuniandCampylobacter coliisolated from hospitalized children, raw chicken, and raw beef in Tehran, Iran

2011 ◽  
Vol 57 (2) ◽  
pp. 143-148 ◽  
Author(s):  
Mohammad Hamidian ◽  
Maryam Sanaei ◽  
Mehdi Bolfion ◽  
Hossein Dabiri ◽  
Mohammad-Reza Zali ◽  
...  
Keyword(s):  
Campylobacter Jejuni ◽  
Rrna Gene ◽  
Campylobacter Coli ◽  
Hospitalized Children ◽  
Unequal Distribution ◽  
Exact Test ◽  
Virulence Markers ◽  
Dnaj Gene ◽  
Different Origins ◽  
Campylobacter Spp

The incidence of the virulence-associated genes cdtA, cdtB, cdtC, cadF, dnaJ, racR, and pldA has been investigated in Campylobacter jejuni and Campylobacter coli collected from raw chicken and beef from retailers in Tehran, Iran, and from hospitalized children (age, ≤14 years) suffering from diarrhea. Campylobacter spp. were collectively identified by morphological and biochemical methods. Campylobacter jejuni and C. coli were discriminated from other Campylobacter spp. by amplification of a specific conserved fragment of the 16S rRNA gene. The distinction between C. jejuni and C. coli was subsequently made by molecular determination of the presence of the hipO gene in C. jejuni or the ask gene in C. coli. Fragments of the studied virulence-associated genes, cdtA, cdtB, cdtC, cadF, racR, dnaJ, and pldA, were amplified by PCR and subjected to horizontal gel electrophoresis. A total of 71 isolates of C. jejuni and 24 isolates of C. coli from meat were analyzed, while the numbers of isolates from the hospitalized children were 28 and 9, respectively. The unequal distribution of C. jejuni and C. coli in the samples has also been reported in other studies. Statistical analyses by the use of the two-tailed Fisher’s exact test of the occurrence of the virulence genes in the isolates of different origins showed that the occurrence of the dnaJ gene was consistently significantly higher in all C. jejuni isolates than in C. coli. The occurrence of the other virulence markers did not differ significantly between species in the majority of the isolates. The PCR results also showed that the occurrence of the virulence markers in the analyzed isolates was much lower than in other studies, which may be caused by a divergent genomic pool of our isolates in comparison with others.

Enrichment Broth for the Detection of Campylobacter jejuni and Campylobacter coli in Fresh Produce and Poultry

2017 ◽  
Vol 80 (11) ◽  
pp. 1842-1850 ◽  
Author(s):  
Youmi Jo ◽  
Hye-Min Oh ◽  
Yohan Yoon ◽  
Sun-Young Lee ◽  
Ji-Hyoung Ha ◽  
...  
Keyword(s):  
Campylobacter Jejuni ◽  
Fresh Produce ◽  
International Organization ◽  
Natural Food ◽  
Campylobacter Coli ◽  
Standardization Method ◽  
Potassium Tellurite ◽  
Enrichment Broth ◽  
International Organization For Standardization ◽  
Campylobacter Spp

ABSTRACT Although campylobacteriosis caused by Campylobacter jejuni and Campylobacter coli has been increasingly reported worldwide owing to the consumption of contaminated poultry and fresh produce, the current detection protocols are not selective enough to inhibit unspecific microbes other than these pathogens. Five antibiotics were separately added to Bolton broth, and the survival rates of 18 Campylobacter spp. and 79 non-Campylobacter spp. were evaluated. The survival rate of the non-Campylobacter spp. was the lowest in Bolton broth with rifampin (6.3%), followed by cefsulodin (12.7%), novobiocin (16.5%), and potassium tellurite and sulfamethozaxole (both 17.7%). Also the most effective concentration of rifampin was found to be 12.5 mg/L, which markedly inhibited non-Campylobacter strains while not affecting the survival of Campylobacter strains. After the Campylobacter spp. were enriched in Bolton broth supplemented with 12.5 mg/L rifampin (R-Bolton broth), CampyFood Agar (CFA) was found to be better in selectively isolating the pathogens in the enrichment broth than the International Organization for Standardization method of using modified charcoal cefoperazone deoxycholate agar (mCCDA) for this step. When applied to natural food samples—here, romaine lettuce, pepper, cherry tomato, Korean leek, and chicken—the R-Bolton broth–CFA combination decreased the number of false-positive results by 50.0, 4.2, 20.8, 50.0, and 94.4%, respectively, compared with the International Organization for Standardization method (Bolton broth–mCCDA combination). These results demonstrate that the combination of R-Bolton broth and CFA is more efficient in detecting C. jejuni and C. coli in poultry and fresh produce and thus should replace the Bolton broth–mCCDA combination.

Production and Characterization of a Monoclonal Antibody to Campylobacter jejuni

2009 ◽  
Vol 72 (4) ◽  
pp. 870-875 ◽  
Author(s):  
S. A. HEO ◽  
R. NANNAPANENI ◽  
M. G. JOHNSON ◽  
J. S. PARK ◽  
K. H. SEO
Keyword(s):  
Monoclonal Antibody ◽  
Campylobacter Jejuni ◽  
Immunosorbent Assay ◽  
Enzyme Linked Immunosorbent Assay ◽  
Campylobacter Coli ◽  
Carbonate Buffer ◽  
Horse Blood ◽  
Microaerobic Conditions ◽  
Immunoglobulin Subclass

Campylobacter species are a group of spiral-shaped bacteria that can cause disease in humans and animals. We developed a high-affinity monoclonal antibody (MAb) probe that recognizes Campylobacter jejuni cells. Cell suspensions grown under microaerobic conditions at 42°C for 20 h on Bolton agar plates with lysed horse blood were used as live and heat-killed preparations, centrifuged at 8,000 × g for 20 min, and resuspended in carbonate buffer (pH 9.6) for coating on the enzyme-linked immunosorbent assay plates. BALB/c mice were immunized with C. jejuni sonicated cells at 107 CFU/ml to generate MAb-producing hybridoma clones. Of about 500 initial hybridoma clones, MAb 33D2, which reacted with C. jejuni and Campylobacter coli, was selected for further evaluation. MAb 33D2 is in the immunoglobulin subclass G2a and had relatively weaker reactivity with the C. coli strains tested. MAb 33D2 did not show any cross-reactions with the nine non-Campylobacter bacteria tested in the enzyme-linked immunosorbent assay and had a stronger affinity for C. jejuni as live versus heat-killed cells. In Western blot assays, MAb 33D2 recognized two major antigens of 62 and 43 kDa in extracts from C. jejuni cells but only one antigen of 62 kDa in extracts from C. coli cells.

scholarly journals Toxin production by Campylobacter spp.

1997 ◽  
Vol 10 (3) ◽  
pp. 466-476 ◽  
Author(s):  
T M Wassenaar
Keyword(s):  
Animal Model ◽  
Campylobacter Jejuni ◽  
Virulence Factors ◽  
Clinical Data ◽  
Related Species ◽  
Toxin Production ◽  
Biochemical Properties ◽  
Future Directions ◽  
Model Studies ◽  
Campylobacter Spp

Of all the virulence factors that were proposed for Campylobacter jejuni and related species to cause disease in humans, the discovery of toxin production was the most promising but led to a rather confusing and even disappointing stream of data. The discussion of whether proteinaceous exotoxins are relevant in disease remains open. One important reason for this lack of consensus is the anecdotal nature of the literature reports. To provide a basis for an unbiased opinion, this review compiles all described exotoxins, compares their reported properties, and provides a summary of animal model studies and clinical data. The toxins are divided into enterotoxins and cytotoxins and are sorted according to their biochemical properties. Since many Campylobacter toxins have been compared with toxins of other species, some key examples of the latter are also discussed. Future directions of toxin research that appear promising are defined.

scholarly journals Isolation, identification, and antimicrobial susceptibility pattern of Campylobacter jejuni and Campylobacter coli from cattle, goat, and chicken meats in Mekelle, Ethiopia

PLoS ONE ◽  
2021 ◽  
Vol 16 (2) ◽  
pp. e0246755
Author(s):  
Yohans Hagos ◽  
Getachew Gugsa ◽  
Nesibu Awol ◽  
Meselu Ahmed ◽  
Yisehak Tsegaye ◽  
...  
Keyword(s):  
Campylobacter Jejuni ◽  
Antibiotic Susceptibility ◽  
Diffusion Method ◽  
Campylobacter Coli ◽  
Susceptibility Pattern ◽  
Isolation And Identification ◽  
Raw Meat ◽  
Antibiotic Susceptibility Test ◽  
Meat Samples ◽  
Campylobacter Spp

Campylobacter jejuni and Campylobacter coli are globally recognized as a major cause of bacterial foodborne gastroenteritis. A cross-sectional study was conducted from October 2015 to May 2016 in Mekelle city to isolate, identify, and estimate the prevalence of C. jejuni and C. coli in raw meat samples and to determine their antibiotic susceptibility pattern. A total of 384 raw meat samples were randomly collected from bovine (n = 210), goat (n = 108), and chicken (n = 66), and isolation and identification of Campylobacter spp. were performed using standard bacteriological techniques and PCR. Antibiotic susceptibility test was performed using disc diffusion method. Of the total 384 raw meat samples, 64 (16.67%) were found positive for Campylobacter spp. The highest prevalence of Campylobacter spp. was found in chicken meat (43.93%) followed by bovine meat (11.90%) and goat meat (9.25%). The most prevalent Campylobacter spp. isolated from meat samples was C. jejuni (81.25%). The overall prevalence of Campylobacter in restaurants, butcher shops, and abattoir was 43.93%, 18.30%, and 9.30%, respectively. 96.8%, 81.25%, 75%, and 71% of the Campylobacter spp. isolates were sensitive to norfloxacin, erythromycin, chloramphenicol, and sulphamethoxazole-trimethoprim, respectively. However, 96.9%, 85.9%, and 50% of the isolates were resistant to ampicillin, amoxicillin, and streptomycin, respectively. Strains that developed multi-drug resistant were 68.7%. The result of this study revealed the occurrence of Campylobacter in bovine, goat, and chicken meats. Hence, there is a chance of acquiring infection via consumption of raw or undercooked meat. Thus, implementation of hygienic practices from a slaughterhouse to the retailers, proper handling and cooking of foods of meat are very important in preventing Campylobacter infection.

scholarly journals Prevalence of Campylobacter among goats and retail goat meat in Congo

2014 ◽  
Vol 8 (02) ◽  
pp. 168-175 ◽  
Author(s):  
Rosette Kabwang A Mpalang ◽  
Raphaël Boreux ◽  
Pierrette Melin ◽  
Khang'Mate Akir Ni Bitiang ◽  
Georges Daube ◽  
...  
Keyword(s):  
Campylobacter Jejuni ◽  
Democratic Republic Of Congo ◽  
Campylobacter Coli ◽  
Sample Collection ◽  
Goat Meat ◽  
Faecal Samples ◽  
Significant Difference ◽  
Meat Goat ◽  
Open Air Markets ◽  
Campylobacter Spp

Background: The prevalence of Campylobacter jejuni and Campylobacter coli was determined in goat and goat meat sold at retail outlets in Lubumbashi, Democratic Republic of Congo (DR Congo). Methodology: A total of 644 samples, including 177 goat meat, 86 goat stomachs, 139 ready to eat (RTE) goat skewers, and 242 goat faecal samples were examined for the presence of Campylobacter jejuni and Campylobacter coli using polymerase chain reaction. Results: Overall, Campylobacter spp. were found in 34.6% of the examined samples. C. jejuni was isolated in 10.1% and C. coli in 26.7% of samples. Only 2.2% of all samples were positive for both species. There was a significant association between the prevalence of C. coli and the type of sample (p < 0.05). The overall prevalence of Campylobacter in different sample groups was 41.2%, 37.2%, 23.7%, and 35.1% for goat meat, goat stomachs, RTE goat skewers, and goat faecal samples, respectively. There was no significant difference (p > 0.05) between the prevalence observed in the rainy season (16.7%) and the dry season (20.0%). Moreover, the overall prevalence of Campylobacter in slaughter sites, open-air markets, warehouses, and semi-open-air markets was 28.2%, 34.2%, 35.4%, and 42.9%, respectively. Statistically, there was no influence of the sample collection site on the frequency of isolation of Campylobacter (p > 0.05). Conclusion: This study shows that, considering the relatively high prevalence of this pathogen, live goat and goat meat are major sources of human and environmental contamination by Campylobacter spp. in Lubumbashi.

Campylobacter jejuni/coli: Methodology of Isolation and Possible Interfering Factors in Primary Culture

1996 ◽  
Vol 59 (4) ◽  
pp. 429-432 ◽  
Author(s):  
MARIA HELENA C. AQUINO ◽  
JOSÉ CARLOS P. CARVALHO ◽  
ANITA TIBANA ◽  
ROBSON M. FRANCO
Keyword(s):  
Primary Culture ◽  
Campylobacter Jejuni ◽  
Campylobacter Coli ◽  
Enrichment Broth ◽  
Direct Plating ◽  
Indicator Microorganisms ◽  
Enrichment Procedure ◽  
Campylobacter Spp

The prevalence of Campylobacter jejuni and Campylobacter coli was investigated in 64 samples of fresh retail chicken purchased from commercial slaughterhouses located in Brazil. Campylobacter spp. were isolated from 40 (62.5%) of 64 analyzed samples. The strains biotyped according to Lior were classified as C. jejuni biotypes I and II, and C. coli biotypes I and II. The efficiency of different procedures for recovering Campylobacter spp. from chicken carcasses was tested. The enrichment procedure was significantly less effective than direct plating (P &lt; 0.05), detecting 19 of 40 (47.5%) as opposed to 38 of 40 (95%) positive samples. Using direct plating the efficiency of Blaser's selective supplement was significantly more effective (P &lt; 0.05) than Skirrow's selective supplement. To verify which factors could be affecting Campylobacter spp. growth in enrichment broth, the pH was measured after incubation for 48 h at 42°C and lactobacilli, coliforms, and enterococci were enumerated. Most of the Campylobacter-negative samples presented high levels of indicator microorganisms, which may have hindered the recovery of Campylobacter spp. during the enrichment procedure.

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