Campylobacter spp. are the leading cause of human gastroenteritis worldwide. Epithelial cell invasion is thought to be essential for Campylobacter spp. infection. Previous invasion studies with intestinal epithelial cells revealed that the ability of different Campylobacter jejuni isolates to inva...
Kuana, Suzete Lora; dos Santos, Luciana Ruschel; Rodrigues, Laura Beatriz; Borsoi, Anderlise; Moraes, Hamilton Luis do Souza; Salle, Carlos Tadeu Pippi; do Nascimento, Vladimir Pinheiro
The aim of this study was to assess the antimicrobial susceptibility of 62 Campylobacter spp. strains obtained from broiler flocks using the agar diffusion method. The Campylobacter spp strains were isolated from 22 flocks aged between 3 and 5 weeks of life, isolated from cloacae swabs, stools and cecal droppings in the farm and from the carcass rinsing in the slaughterhouse. Campylobacter spp strains were tested on Mueller-Hilton (MH) agar (27 samples) and MH plus TTC agar (35 samples). The antimicrobial susceptibility test revealed a 62.5% resistance to at least one drug, especially to enrofloxacin (71%), neomycin (50%), lincomycin (50%), tetracycline (43%), penicillin (42%), ceftiofur (33%) amoxicillin (27%), spiramycin (20%), ampicillin (18%) and norfloxacin (14%), whereas a lower percentage of strains was resistant to erythromycin (10%) and doxycycline (10%). All strains were sensitive to gentamicin and lincomycin-spectinomycin and 80% of them to colistin. These results indicate that it is necessary to reduce the use of antimicrobials in veterinary and human medicine. PMID:24031299
Kuana, Suzete Lora; Dos Santos, Luciana Ruschel; Rodrigues, Laura Beatriz; Borsoi, Anderlise; Moraes, Hamilton Luis do Souza; Salle, Carlos Tadeu Pippi; do Nascimento, Vladimir Pinheiro
The aim of this study was to assess the antimicrobial susceptibility of 62 Campylobacter spp. strains obtained from broiler flocks using the agar diffusion method. The Campylobacter spp strains were isolated from 22 flocks aged between 3 and 5 weeks of life, isolated from cloacae swabs, stools and cecal droppings in the farm and from the carcass rinsing in the slaughterhouse. Campylobacter spp strains were tested on Mueller-Hilton (MH) agar (27 samples) and MH plus TTC agar (35 samples). The antimicrobial susceptibility test revealed a 62.5% resistance to at least one drug, especially to enrofloxacin (71%), neomycin (50%), lincomycin (50%), tetracycline (43%), penicillin (42%), ceftiofur (33%) amoxicillin (27%), spiramycin (20%), ampicillin (18%) and norfloxacin (14%), whereas a lower percentage of strains was resistant to erythromycin (10%) and doxycycline (10%). All strains were sensitive to gentamicin and lincomycin-spectinomycin and 80% of them to colistin. These results indicate that it is necessary to reduce the use of antimicrobials in veterinary and human medicine.
García-Peña, F J; Pérez-Boto, D; Jiménez, C; San Miguel, E; Echeita, A; Rengifo-Herrera, C; García-Párraga, D; Ortega-Mora, L M; Pedraza-Díaz, S
The presence of Campylobacter spp. was investigated in 41 Antarctic fur seals (Arctocephalus gazella) and 9 Weddell seals (Leptonychotes weddellii) at Deception Island, Antarctica. Infections were encountered in six Antarctic fur seals. The isolates, the first reported from marine mammals in the Antarctic region, were identified as Campylobacter insulaenigrae and Campylobacter lari.
Humski, Andrea; Njari, Bela; Ostović, Mario; Duvnjak, Sanja; Cvetnić, Željko
Summary In order to detect thermotolerant Campylobacter spp., 241 samples of fresh chicken meat, at retail in Croatia, were analysed according to a standard method, followed by biochemical test and molecular polymerase chain reaction/restriction enzyme analysis for exact species determination. Campylobacter spp. prevalence was 73.86%. Campylobacter jejuni and Campylobacter coli were isolated from 53.53 and 15.35% of the samples, respectively. In 4.98% of isolates thermotolerant Campylobacter spp. were not determined. The multilocus sequence typing method was used to evaluate genetic diversity of eight Campylobacter jejuni and four Campylobacter coli isolates. To our knowledge, these results of genotyping provided the first data on the presence of sequence types (STs) and clonal complexes (CCs) of Campylobacter jejuni and C. coli isolates in Croatia. By applying the multilocus sequence typing, a new allele of tkt gene locus was discovered and marked tkt508. The C. jejuni ST 6182 and C. coli ST 6183 genotypes were described for the first time, and all other identified genotypes were clustered in the previously described sequence types and clonal complexes. These findings provide useful information on the prevalence and epidemiology of Campylobacter jejuni and C. coli in Croatia. PMID:28115906
Varela, Norma P.; Friendship, Robert M.; Dewey, Cate E.
This study aimed to establish the prevalence of Campylobacter spp. in 80 Ontario grower-finisher pig herds. Ninety-nine percent of the isolates yielded Campylobacter, C. coli being the most common species detected. Control of this microorganism must rely on careful food processing and storage of pork, rather than on an on-farm approach. PMID:17542372
Fraqueza, M J; Martins, A; Borges, A C; Fernandes, M H; Fernandes, M J; Vaz, Y; Bessa, R J B; Barreto, A S
The aim of the current work was to evaluate the prevalence and antimicrobial susceptibility of Campylobacter spp. isolated from different chicken production systems at the slaughterhouse level. Chicken sampling at slaughterhouse was performed for cecum, carcass, and breast meat from flocks of organic (n = 6), extensive indoor (n = 14), and intensive production (n = 14), totaling 34 ceca pools, 64 neck skin pools, and 132 breasts, representing 96,386 chickens. A collection of 167 strains were identified as Campylobacter coli (n = 85) and Campylobacter jejuni (n = 82) and were tested for susceptibility to 11 antimicrobial agents by the disk diffusion method. The frequency of Campylobacter in chicken samples from different production systems was between 79 and 100%. Campylobacter isolated from all origins were resistant to the fluoroquinolones studied (80-98%). However, for ciprofloxacin and ofloxacin, the Campylobacter isolates from extensive indoor chicken were significantly (P < 0.05) less resistant (77 and 58%) than that from organic (97 and 91%) and intensive production (96 and 95%). A high probability of tetracycline resistance occurrence was also found for the Campylobacter spp. tested (58% for C. jejuni and 76% for C. coli). A more frequent profile of multidrug resistance was noticed for isolates from intensive and organic production than for extensive indoor production. These results reinforce the need of efficient strategy implementation to control and reduce Campylobacter in chickens at production and slaughter levels, and the necessity to reduce the use of antimicrobials in poultry sector.
Stone, Diana; Davis, Margaret; Baker, Katherine; Besser, Tom; Roopnarine, Rohini; Sharma, Ravindra
This study determined whether multilocus sequence types (MLST) of Campylobacter from poultry in 2 farms in Grenada, West Indies, differed by farm, antimicrobial resistance and farm antibiotic use. Farm A used fluoroquinolones in the water and Farm B used tetracyclines. The E-test was used to determine resistance of isolates to seven antibiotics. PCR of the IpxA gene confirmed species and MLST was used to characterize 38 isolates. All isolates were either C. jejuni or C. coli. Farm antibiotic use directly correlated with antimicrobial resistance of Campylobacter isolates. Almost 80% of the isolates from Farm A were fluoroquinolone resistant and 17.9% of the isolates from Farm B were fluoroquinolone resistant. All Campylobacter isolates from Farm A were tetracycline sensitive, whereas 35.7% of isolates from Farm B were tetracycline resistant. Six previously recognized sequence types (STs) and 2 novel STs were identified. Previously recognized STs were those overwhelmingly reported from poultry and humans globally. Isolates with the same ST did not always have the same antibiotic resistance profile. There was little ST overlap between the farms suggesting that within-farm transmission of Campylobacter genotypes may dominate. MLST typing was useful for tracking Campylobacter spp. among poultry units and can help elucidate Campylobacter host-species population structure and its relevance to human health.
Dipineto, Ludovico; De Luca Bossa, Luigi Maria; Russo, Tamara Pasqualina; Cutino, Eridania Annalisa; Gargiulo, Antonio; Ciccarelli, Francesca; Raia, Pasquale; Menna, Lucia Francesca; Fioretti, Alessandro
A total of 170 birds of prey admitted to two Wildlife Rescue and Rehabilitation Centers of Italy were examined. Birds were divided by diurnal (n = 15) and nocturnal (n = 7) species, sampled by cloacal swabs, and examined for Campylobacter spp. by cultural and molecular methods. Campylobacter spp. were isolated in 43 out of the 170 (25.3%) birds of prey examined. Among these, 43/43 (100%) were identified as Campylobacter jejuni and 10/43 (23.3%) were identified as Campylobacter coli recovered from mixed infections. Diurnal birds of prey showed a significantly higher prevalence value (P = 0.0006) for Campylobacter spp. than did nocturnal birds of prey.
Ku, Bok Kyung; Kim, Hae Ji; Lee, Young Ju; Kim, Young Ihl; Choi, Jung Su; Park, Mi Young; Kwon, Jin Wook; Nam, Hyang-Mi; Kim, Yong Hwan; Jung, Suk-Chan; Lee, Sun Jin; Kim, Sang Hyun; Kim, Jong Hyun
This study was conducted to examine the in vitro activity of antimicrobials against Campylobacter spp. isolates from chicken and human sources and the genetic interrelation among them. During 2004-2008, a total of 173 Campylobacter spp. isolated from chicken meats (60 domestic and 62 imported chicken meats) and humans (n = 51) were tested for susceptibility to nine antimicrobials. Of 173 isolates, 140 (80.9%) showed multidrug resistance (MDR) against three to eight antimicrobials. The most frequent pattern type was MDR to four antimicrobials: ciprofloxacin, nalidixic acid, ampicillin, and tetracycline. Over 52.6% (91/173) of the isolates tested were resistant to these four antibiotics simultaneously. Especially, two and five isolates originated from Korea and Brazil showed resistance against all antibiotics tested, except for florfenicol. Further, 95% (57/60) of the isolates originated from domestic chicken showed resistance to ciprofloxacin, the antimicrobial agent of choice for treatment of human campylobacteriosis. Genotypic characterization of all Campylobacter isolates performed by pulsed-field gel electrophoresis yielded 74 types among the 173 isolates. Isolates sharing the same or similar genetic clusters were detected in different countries at different times. The pulsed-field gel electrophoresis patterns of chicken-related isolates were closely related to those of isolates from humans with gastroenteritidis. The results of this study suggest that MDR Campylobacter spp. are widespread and that Campylobacter with similar genotypes are circulating both in humans and in chicken meat in Korea.
Sato, K.; Bartlett, P. C.; Kaneene, J. B.; Downes, F. P.
The prevalence and antimicrobial susceptibilities of Campylobacter spp. isolates from bovine feces were compared between organic and conventional dairy herds. Thirty organic dairy herds, where antimicrobials are rarely used for calves and never used for cows, were compared with 30 neighboring conventional dairy farms, where antimicrobials were routinely used for animals for all ages. Fecal specimens from 10 cows and 10 calves on 120 farm visits yielded 332 Campylobacter isolates. The prevalence of Campylobacter spp. in organic and conventional farms was 26.7 and 29.1%, and the prevalence was not statistically different between the two types of farms. Campylobacter prevalence was significantly higher in March than in September, higher in calves than in cows, and higher in smaller farms than in large farms. The rates of retained placenta, pneumonia, mastitis, and abortion were associated with the proportion of Campylobacter isolation from fecal samples. The gradient disk diffusion MIC method (Etest) was used for testing susceptibility to four antimicrobial agents: ciprofloxacin, gentamicin, erythromycin, and tetracycline. Two isolates were resistant to ciprofloxacin, and none of isolates was resistant to gentamicin or erythromycin. Resistance to tetracycline was 45% (148 of 332 isolates). Tetracycline resistance was found more frequently in calves than in cows (P = 0.042), but no difference was observed between organic and conventional farms. When we used Campylobacter spp. as indicator bacteria, we saw no evidence that restriction of antimicrobial use on dairy farms was associated with prevalence of resistance to ciprofloxacin, gentamicin, erythromycin, and tetracycline. PMID:15006764
Sato, K; Bartlett, P C; Kaneene, J B; Downes, F P
The prevalence and antimicrobial susceptibilities of Campylobacter spp. isolates from bovine feces were compared between organic and conventional dairy herds. Thirty organic dairy herds, where antimicrobials are rarely used for calves and never used for cows, were compared with 30 neighboring conventional dairy farms, where antimicrobials were routinely used for animals for all ages. Fecal specimens from 10 cows and 10 calves on 120 farm visits yielded 332 Campylobacter isolates. The prevalence of Campylobacter spp. in organic and conventional farms was 26.7 and 29.1%, and the prevalence was not statistically different between the two types of farms. Campylobacter prevalence was significantly higher in March than in September, higher in calves than in cows, and higher in smaller farms than in large farms. The rates of retained placenta, pneumonia, mastitis, and abortion were associated with the proportion of Campylobacter isolation from fecal samples. The gradient disk diffusion MIC method (Etest) was used for testing susceptibility to four antimicrobial agents: ciprofloxacin, gentamicin, erythromycin, and tetracycline. Two isolates were resistant to ciprofloxacin, and none of isolates was resistant to gentamicin or erythromycin. Resistance to tetracycline was 45% (148 of 332 isolates). Tetracycline resistance was found more frequently in calves than in cows (P = 0.042), but no difference was observed between organic and conventional farms. When we used Campylobacter spp. as indicator bacteria, we saw no evidence that restriction of antimicrobial use on dairy farms was associated with prevalence of resistance to ciprofloxacin, gentamicin, erythromycin, and tetracycline.
Scott, L; Menzies, P; Reid-Smith, R J; Avery, B P; McEwen, S A; Moon, C S; Berke, O
The objectives of this study were to determine the prevalence of antimicrobial resistance (AMR) in faecal Campylobacter spp. from lambs and adult sheep and associations between antimicrobial use (AMU) and AMR. A total of 275 faecal samples collected during initial and final visits from 51 sheep flocks, including one feedlot, across southern Ontario were tested for the presence of Campylobacter spp. Campylobacter jejuni was detected in 52% (143/275) of the faecal samples, Campylobacter coli in 7% (19/275), Campylobacter lari in 1% (2/275) and 2% (4/275) were non-speciated Campylobacter. Broth microdilution was used to test antimicrobial susceptibility of 162 isolates to nine antimicrobials. Campylobacter jejuni isolates (n = 142) were resistant to tetracycline (39%), ciprofloxacin (4%), nalidixic acid (4%) and telithromycin (1%). C. coli isolates (n = 19) were resistant to tetracycline (74%), and azithromycin, clindamycin, erythromycin, and telithromycin (5%). The C. lari isolate displayed resistance to nalidixic acid. No statistically significant associations were found between AMU and AMR during multivariate modelling in this study.
García-Peña, F J; Llorente, M T; Serrano, T; Ruano, M J; Belliure, J; Benzal, J; Herrera-León, S; Vidal, V; D'Amico, V; Pérez-Boto, D; Barbosa, A
The presence of Campylobacter species was studied in three Antarctic penguin species, Adélie (Pygoscelis adeliae), chinstrap (Pygoscelis antarctica) and gentoo (Pygoscelis papua). A total of 390 penguins were captured in 12 different rookeries along the Antarctic Peninsula with differences in the amount of human visitation: six colonies were highly visited [Stranger Point, King George Island (P. papua and P. adeliae); Hannah Point, Livingston Island (P. papua and P. antarctica); Deception Island (P. antarctica); and Paradise Bay, Antarctic Peninsula (P. papua)], and six colonies were rarely visited [Devil's Point, Byers Peninsula, Livingston Island (P. papua); Cierva Cove, Antarctic Peninsula (P. papua); Rongé Island (P. papua and P. antarctica); Yalour Island (P. adeliae); and Avian Island (P. adeliae)]. A total of 23 strains were isolated from penguins from nine different rookeries. Campylobacter lari subsp. lari was isolated from eight samples (seven from P. papua and one from P. adeliae); C. lari subsp. concheus from 13 (ten from P. adeliae and three from P. antarctica) and C. volucris from two samples (both from P. papua). We did not find any significant differences in the prevalence of Campylobacter spp. between the populations in highly and rarely visited areas. This is the first report of C. lari subsp. concheus and C. volucris isolation from penguins in the Antarctic region.
Deckert, Anne; Valdivieso-Garcia, Alfonso; Reid-Smith, Richard; Tamblyn, Susan; Seliske, Patrick; Irwin, Rebecca; Dewey, Cate; Boerlin, Patrick; McEwen, Scott A
Campylobacter is an important enteric pathogen of humans and can cause diarrhea, fever, and abdominal pain. Campylobacter infections have frequently been associated with the handling and consumption of raw and undercooked poultry. Antimicrobial resistance among Campylobacter strains is of concern in the treatment of campylobacteriosis in vulnerable populations. A 2-year multidisciplinary study was conducted in the Perth and Wellington-Dufferin-Guelph public health units in Ontario, Canada, to investigate the prevalence and antimicrobial resistance of Campylobacter spp. in retail chicken. Retail chicken samples were collected from randomly selected stores in these health units. Resulting Campylobacter isolates were tested for susceptibility to amoxicillin-clavulanic acid (AMC), ampicillin (AMP), chloramphenicol (CHL), ciprofloxacin (CIP), clindamycin (CLI), erythromycin (ERY), gentamicin (GEN), nalidixic acid (NAL), tetracycline (TCY), and trimethoprim-sulfamethoxazole (SXT) using the E test. The prevalence of Campylobacter in 1,256 retail chicken samples was 59.6%. Of these positive samples, 9% contained Campylobacter coli, 1% contained Campylobacter lari, and 90% contained Campylobacter jejuni. Of the chicken isolates that were resistant to one or more antimicrobial agents, 301 isolates (40%) were resistant to one agent, 374 (50%) were resistant to two, 39 (5%) were resistant to three, 20 (3%) were resistant to four, and 6 (1%) were resistant to five. Nine isolates (1%) were susceptible to all antimicrobial agents tested. All isolates were susceptible to AMC, CHL, and GEN. Less than 10% of isolates were resistant to NAL, CIP, CLI, ERY, and AMP. Resistance to TCY was common (56%). No isolates had a resistance pattern that included all three antimicrobials important in the treatment of human campylobacteriosis (CIP, ERY, and TCY); however, 24 isolates (3.2%) were resistant to at least two of these antimicrobials.
Frediani-Wolf, V; Stephan, R
The aim of this study was to determine resistance patterns of strains of Campylobacter spp. isolated from poultry carcasses in one of the two big Swiss poultry slaughterhouses. A variety of antibiotics with clinical relevance in human and/or in veterinary medicine was tested. In addition, the results of the disc diffusion method, E-test and microdilution broth methods were compared. Of the 195 Campylobacter jejuni strains isolated from 195 poultry carcasses from 21 flocks, 134 strains were susceptible in vitro to all tested antibiotics. Sixty-one strains (31.3%, from eight flocks) showed resistance. Forty-one strains were resistant to a single antibiotic-34 to streptomycin, 6 to ampicillin and 1 to ciprofloxacin. Eighteen strains (from two flocks) showed combined resistance to erythromycin and streptomycin, two strains to ciprofloxacin and streptomycin. None of the isolates was resistant to tetracycline. The data of this first study in Switzerland show a favourable resistance situation for C. jejuni strains against erythromycin, tetracycline and ciprofloxacin. The disc diffusion method was found to be a reliable and easy tool for monitoring the prevalence of resistant C. jejuni strains. For surveillance of changes in the susceptibility concentration levels to antimicrobial agents, however, a MIC method should be used. Further investigations along the whole poultry production chain (farm, slaughterhouse and retail levels) are now necessary in order to confirm the resistance situation.
Tamborini, Ana L; Casabona, Luis M; Viñas, María R; Asato, Valeria; Hoffer, Alicia; Farace, María I; Lucero, María C; Corso, Alejandra; Pichel, Mariana
The prevalence of Campylobacter spp. was investigated in 327 patients suffering from diarrhea and in 36 animals (dogs, cats and chickens) owned by the patients that presented infection by Campylobacter in Santa Rosa, La Pampa, Argentina. Campylobacter spp. was isolated in 50/327 patients and in 12/36 animals, being Campylobacter jejuni the most common species. Resistance to ciprofloxacin (65 %) and tetracycline (32 %) was found among 35 isolates of human origin studied. Seven genetic subtypes were observed among 13 C. jejuni isolates by pulsed field gel electrophoresis. Two subtypes grouped isolates belonging to patients and their respective dogs whereas another subtype grouped one isolate of human origin and two isolates from the patient's chickens. The results of this investigation highlight the need to strengthen surveillance of Campylobacter spp. not only in poultry, which is recognized as the main reservoir, but also in pets, which were shown to be asymptomatic carriers of the pathogen.
Bojanić, K; Midwinter, A C; Marshall, J C; Rogers, L E; Biggs, P J; Acke, E
Campylobacter causes acute gastroenteritis in people worldwide and is frequently isolated from food, animals and the environment. The disease is predominately food-borne but many routes of transmission and sources of infection have been described, including contact with pets. The prevalence of Campylobacter spp. in dogs and cats varies widely, and data on New Zealand pets are limited. This study aimed to investigate the prevalence of Campylobacter spp. in dogs, cats and retail raw meat pet food products in New Zealand and to characterize Campylobacter jejuni isolates using multilocus sequence typing (MLST). Ninety dogs and 110 cats examined at the Massey University Veterinary Teaching Hospital for elective procedures, and fifty locally purchased retail raw meat pet diets were sampled. Two culture protocols combining Bolton broth enrichment and mCCDA and CAT agars in a microaerobic atmosphere at 42°C and 37°C with species identification using PCR were performed. The prevalence of Campylobacter spp., C. jejuni, Campylobacter upsaliensis and Campylobacter helveticus was 36%, 13%, 23% and 1% in dogs and 16%, 5%, 5% and 7% in cats, respectively. One dog had Campylobacter lari confirmed, and three dogs and one cat had multiple Campylobacter spp. detected. Significantly more animals tested positive using CAT than mCCDA agar (P < 0.001). Being neutered, vaccinated for Bordetella bronchiseptica, fed dry diets and brought in for neutering were protective factors for dogs, whereas attendance for dental treatment was a risk factor for cats. Campylobacter spp. were isolated from 28%, C. jejuni 22%, C. lari 6% and Campylobacter coli 6% of food samples. Six isolates positive by Campylobacter genus PCR were identified as Arcobacter butzleri. Poultry meat was more likely to be positive than non-poultry meat (P = 0.006). Of the 13 C. jejuni pet isolates with full MLST profiles, eight were of different sequence types (ST) and all nine food isolates were of different STs.
Santaniello, Antonio; Dipineto, Ludovico; Veneziano, Vincenzo; Mariani, Ugo; Fioretti, Alessandro; Menna, Lucia Francesca
Thermotolerant Campylobacter spp. were isolated from 118/240 (49.2%) rectal swabs from commercially farmed hares (Lepus europaeus) in southern Italy. Using multiplex PCR, Campylobacter coli was identified in 118/118 (100%) positive samples, while 17/118 (14.4%) positive samples were also positive for Campylobacter jejuni. Adult hares had a higher prevalence of infection with Campylobacter spp. than juvenile hares.
Wieczorek, Kinga; Kania, Iwona; Osek, Jacek
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.
Duarte, Andreia; Santos, Andrea; Manageiro, Vera; Martins, Ana; Fraqueza, Maria J; Caniça, Manuela; Domingues, Fernanda C; Oleastro, Mónica
Infections by Campylobacter jejuni and Campylobacter coli are considered the major cause of bacterial gastroenteritis in humans, with food being the main source of infection. In this study, a total of 196 Campylobacter strains (125 isolates from humans, 39 from retail food and 32 from food animal sources) isolated in Portugal between 2009 and 2012 were characterised by multilocus sequence typing (MLST) and flaA short variable region (SVR) typing. Susceptibility to six antibiotics as well as the mechanisms underlying antibiotic resistance phenotypes was also studied. Based on MLST typing, C. coli strains were genetically more conserved, with a predominant clonal complex (CC828), than C. jejuni strains. In contrast, C. coli isolates were genetically more variable than C. jejuni with regard to flaA-SVR typing. A high rate of resistance was observed for quinolones (100% to nalidixic acid, >90% to ciprofloxacin) and, in general, resistance was more common among C. coli, especially for erythromycin (40.2% vs. 6.7%). In addition, most isolates (86%) were resistant to multiple antimicrobial families. Besides the expected point mutations associated with antibiotic resistance, detected polymorphisms in the cmeABC locus likely play a role in the multiresistant phenotype. This study provides for the first time an overview of the genetic diversity of Campylobacter strains from Portugal. It also shows a worrying antibiotic multiresistance rate and the emergence of Campylobacter strains resistant to antibiotics of human use.
Bakhshi, B.; Kalantar, M.; Rastegar-Lari, A.; Fallah, F.
A total of 70 samples were collected from chicken meat obtained from 10 markets in Tehran, Iran from which 39 Campylobacter coli were isolated. Among 10 antibiotics used, maximum resistance was seen to trimethoprim-sulphamethoxazole (SXT) (97.36%), nalidixic acid (94.8%), ciprofloxacin (87.7%), streptomycin (89.72%), and tetracycline (97.4%). No resistance was to gentamycin was observed. None of the Campylobacter strains under study harbored integron, suggesting the involvement of other resistance mechanisms in emergence of multi drug resistance (MDR) phenotype among the isolates. Two major types (A and B) and 15 subtypes (A1-A8 and B1-B7) were identified. Pulsed-field gel electrophoresis (PFGE) analysis demonstrated a high degree of homogeneity while the majority of the isolates shared identical or very similar PFGE genotypes. Isolates with identical genotypes differed in their resistance profile, although all of them assigned to MDR phenotype. To our knowledge, this is the first molecular survey from Iran characterizing Campylobacter isolates from poultry, which adds to our knowledge the epidemiological linkage of Campylobacter isolates with MDR properties from different sources and emphasizes the need for cautious use of antimicrobials in different fields of food production chain. PMID:27822247
Carrillo, Catherine D; Kenwell, Robyn; Iugovaz, Irene; Oyarzabal, Omar A
The recovery of Campylobacter species from food and environmental sources is challenging due to the slow growth of these bacteria and the need to suppress competing organisms during the isolation procedures. The addition of multiple selective antimicrobials to growth media can negatively impact recovery of some Campylobacter spp. Here, we describe our current method for the isolation of thermotolerant Campylobacter species, mainly C. jejuni and C. coli, from food and environmental samples. We emphasize the use of membrane filtration during plating for the specific isolation of Campylobacter spp. and a reduced use of antimicrobial supplements throughout the whole isolation process.
Yoo, Jin-Hee; Choi, Na-Young; Bae, Young-Min; Lee, Jung-Su; Lee, Sun-Young
This study was conducted to develop a selective medium for the detection of Campylobacter spp. in fresh produce. Campylobacter spp. (n=4), non-Campylobacter (showing positive results on Campylobacter selective agar) strains (n=49) isolated from fresh produce, indicator bacteria (n=13), and spoilage bacteria isolated from fresh produce (n=15) were plated on four Campylobacter selective media. Bolton agar and modified charcoal cefoperazone deoxycholate agar (mCCDA) exhibited higher sensitivity for Campylobacter spp. than did Preston agar and Hunt agar, although certain non-Campylobacter strains isolated from fresh produce by using a selective agar isolation method, were still able to grow on Bolton agar and mCCDA. To inhibit the growth of non-Campylobacter strains, Bolton agar and mCCDA were supplemented with 5 antibiotics (rifampicin, polymyxin B, sodium metabisulfite, sodium pyruvate, ferrous sulfate) and the growth of Campylobacter spp. (n=7) and non-Campylobacter strains (n=44) was evaluated. Although Bolton agar supplemented with rifampicin (BR agar) exhibited a higher selectivity for Campylobacter spp. than did mCCDA supplemented with antibiotics, certain non-Campylobacter strains were still able to grow on BR agar (18.8%). When BR agar with various concentrations of sulfamethoxazole-trimethoprim were tested with Campylobacter spp. (n=8) and non-Campylobacter (n=7), sulfamethoxazole-trimethoprim was inhibitory against 3 of 7 non-Campylobacter strains. Finally, we validated the use of BR agar containing 50mg/L sulfamethoxazole (BRS agar) or 0.5mg/L ciprofloxacin (BRCS agar) and other selective agars for the detection of Campylobacter spp. in chicken and fresh produce. All chicken samples were positive for Campylobacter spp. when tested on mCCDA, BR agar, and BRS agar. In fresh produce samples, BRS agar exhibited the highest selectivity for Campylobacter spp., demonstrating its suitability for the detection of Campylobacter spp. in fresh produce.
Ferro, I D; Benetti, T M; Oliveira, T C R M; Abrahão, W M; Farah, S M S S; Luciano, F B; Macedo, R E F
1. The aim of the present study was to evaluate the antimicrobial resistance of Campylobacter strains (C. jejuni, C. coli and C. lari) isolated from broiler carcasses processed in the State of Paraná, Brazil. 2. Rates of microbial resistance and susceptibility were assessed by both Disk Diffusion (DD) and Etest (Minimum Inhibitory Concentration) techniques. Antibiotics were tested using DD (12 antibiotics) and/or MIC (7 antibiotics) methods. 3. A total of 95.8% of the strains were resistant to at least two agents. In terms of multidrug resistance, 75% of strains were resistant to three or more groups of antibiotics. The highest rates of resistance were detected for cefalotin, ciprofloxacin, tetracycline and nalidixic acid. A high rate of susceptibility of the strains to erythromycin (95.8%) was found confirming that this is considered the agent of choice for treating campylobacteriosis. Comparison of the microbial resistance and susceptibility, as determined simultaneously by the two methods, found the techniques to be statistically equivalent for 5 out of the 6 antibiotics tested. 4. The results of this study suggest the need for adopting measures to control the use of antibiotics in broiler production to prevent multidrug resistance of Campylobacter strains and reduce the risk of serious human diseases caused by the consumption of contaminated chicken meat.
Patrick, Mary; Gonzalez, Anthony; Akin, Joshua; Polage, Christopher R.; Wymore, Kate; Gillim-Ross, Laura; Xavier, Karen; Sadlowski, Jennifer; Monahan, Jan; Hurd, Sharon; Dahlberg, Suzanne; Jerris, Robert; Watson, Renee; Santovenia, Monica; Mitchell, David; Harrison, Cassandra; Tobin-D'Angelo, Melissa; DeMartino, Mary; Pentella, Michael; Razeq, Jafar; Leonard, Celere; Jung, Carrianne; Achong-Bowe, Ria; Evans, Yaaqobah; Jain, Damini; Juni, Billie; Leano, Fe; Robinson, Trisha; Smith, Kirk; Gittelman, Rachel M.; Garrigan, Charles; Nachamkin, Irving
The use of culture-independent diagnostic tests (CIDTs), such as stool antigen tests, as standalone tests for the detection of Campylobacter in stool is increasing. We conducted a prospective, multicenter study to evaluate the performance of stool antigen CIDTs compared to culture and PCR for Campylobacter detection. Between July and October 2010, we tested 2,767 stool specimens from patients with gastrointestinal illness with the following methods: four types of Campylobacter selective media, four commercial stool antigen assays, and a commercial PCR assay. Illnesses from which specimens were positive by one or more culture media or at least one CIDT and PCR were designated “cases.” A total of 95 specimens (3.4%) met the case definition. The stool antigen CIDTs ranged from 79.6% to 87.6% in sensitivity, 95.9 to 99.5% in specificity, and 41.3 to 84.3% in positive predictive value. Culture alone detected 80/89 (89.9% sensitivity) Campylobacter jejuni/Campylobacter coli-positive cases. Of the 209 noncases that were positive by at least one CIDT, only one (0.48%) was positive by all four stool antigen tests, and 73% were positive by just one stool antigen test. The questionable relevance of unconfirmed positive stool antigen CIDT results was supported by the finding that noncases were less likely than cases to have gastrointestinal symptoms. Thus, while the tests were convenient to use, the sensitivity, specificity, and positive predictive value of Campylobacter stool antigen tests were highly variable. Given the relatively low incidence of Campylobacter disease and the generally poor diagnostic test characteristics, this study calls into question the use of commercially available stool antigen CIDTs as standalone tests for direct detection of Campylobacter in stool. PMID:26962088
Repérant, E; Laisney, M J; Nagard, B; Quesne, S; Rouxel, S; Le Gall, F; Chemaly, M; Denis, M
Investigating Campylobacter epidemiology requires adequate technique and media to ensure optimal culturing and accurate detection and isolation of Campylobacter strains. In the present study, we investigated the performances of three enrichment durations in Bolton broth (0, 24 and 48h) and compared four isolation media (mCCDA, Karmali, Butzler no. 2 and CampyFood agar (CFA)) for the detection of Campylobacter positive samples and the identification of Campylobacter species, from naturally contaminated broiler chicken samples (caeca, neck skin from carcasses, and skin from thighs). We compared our local results to those we obtained with samples from a European survey (caeca and neck skin) and a national survey (neck skin, thigh skin, and breast). Direct plating favored the detection of positive samples highly contaminated by Campylobacter (caeca and neck skin from carcasses) whatever the media. A longer enrichment reduced the rates of Campylobacter recovery except when using Butzler no. 2, more particularly for neck skin which background microflora was less important than in caeca. As a matter of fact, enrichment allowed a higher detection rate of positive samples with low Campylobacter contamination levels (breast, thigh skin), this detection being enhanced when using Butzler no. 2. When comparing the 3 other selective media, CFA was the 2nd most efficient media prior to mCCDA and Karmali. Interestingly, enrichment promoted the growth of Campylobacter coli but this promotion was least with Butzler no. 2 agar. Our study has confirmed the need to adapt the method to the types of samples for improving the detection of Campylobacter and that the method may affect the prevalence of the species.
Ingresa-Capaccioni, S; González-Bodí, S; Jiménez-Trigos, E; Marco-Jiménez, F; Catalá, P; Vega, S; Marin, C
Campylobacter is the most common bacterial cause of human gastrointestinal disease in most developed countries. It is generally accepted that poultry products are a significant source of foodborne Campylobacter infections in humans. Assessing the effectiveness of any potential intervention at farm level requires monitoring of the Campylobacter status of broiler flocks, using appropriate sampling methods. The aim of this study was to assess the influence of the sample type across the rearing period for the detection of Campylobacter spp. at farm level. During this study, 21 commercial broiler farms were intensively sampled. Each farm was visited and sampled at different times during the rearing period (d 1, 7, 14, 21, 28, 35, and 42). On the first day of rearing, the status of the house and the day-old flock was evaluated, and environmental and cecal samples were collected. During rearing, 4 different sample types were collected: feces with sock swabs (sock swabs), feces directly from the litter (feces), cloacal swabs, and cecal content. All samples were analyzed according to ISO 10272-1:2006 (Annex E) and also by direct culture. The results of this study showed that Campylobacter spp. were detected in all of the sample types on d 14 of rearing. From this point on, the detection increased significantly, with a maximum detection rate by the end of rearing, regardless of the sample type. All samples that were negative upon direct culture were also negative after pre-enrichment. At the end of rearing, the percentage of samples positive for Campylobacter spp. was 71.4% for cecal samples, 61.9% for cloacal swabs, 45.2% for sock swabs, and 69.1% for fecal samples. C. jejuni was detected in all the sample types, with positive rates ranging from 67.1 to 76.0% for cecal samples and cloacal content, respectively. Cecal samples, cloacal swabs, and fecal samples cultured by direct plating onto modified charcoal cefoperazone deoxycholate agar (mCCDA) without pre-enrichment have
Viswanathan, M; Pearl, D L; Taboada, E N; Parmley, E J; Mutschall, S; Jardine, C M
The objectives of this study were to (i) compare the carriage of Campylobacter and antimicrobial-resistant Campylobacter among livestock and mammalian wildlife on Ontario farms, and (ii) investigate the potential sharing of Campylobacter subtypes between livestock and wildlife. Using data collected from a cross-sectional study of 25 farms in 2010, we assessed associations, using mixed logistic regression models, between Campylobacter and antimicrobial-resistant Campylobacter carriage and the following explanatory variables: animal species (beef, dairy, swine, raccoon, other), farm type (swine, beef, dairy), type of sample (livestock or wildlife) and Campylobacter species (jejuni, coli, other). Models included a random effect to account for clustering by farm where samples were collected. Samples were subtyped using a Campylobacter-specific 40 gene comparative fingerprinting assay. A total of 92 livestock and 107 wildlife faecal samples were collected, and 72% and 27% tested positive for Campylobacter, respectively. Pooled faecal samples from livestock were significantly more likely to test positive for Campylobacter than wildlife samples. Relative to dairy cattle, pig samples were at significantly increased odds of testing positive for Campylobacter. The odds of isolating Campylobacter jejuni from beef cattle samples were significantly greater compared to dairy cattle and raccoon samples. Fifty unique subtypes of Campylobacter were identified, and only one subtype was found in both wildlife and livestock samples. Livestock Campylobacter isolates were significantly more likely to exhibit antimicrobial resistance (AMR) compared to wildlife Campylobacter isolates. Campylobacter jejuni was more likely to exhibit AMR when compared to C. coli. However, C. jejuni isolates were only resistant to tetracycline, and C. coli isolates exhibited multidrug resistance patterns. Based on differences in prevalence of Campylobacter spp. and resistant Campylobacter between
Campylobacter jejuni and C. coli are the two important species responsible for most of the Campylobacter infections in humans. Reliable isolation and detection of Campylobacter spp. from food samples are challenging due to the interferences from complex food substances and the fastidious growth requ...
Tadesse, Daniel A; Bahnson, Peter B; Funk, Julie A; Thakur, Siddhartha; Morrow, William E Morgan; Wittum, Thomas; DeGraves, Fred; Rajala-Schultz, Paivi; Gebreyes, Wondwossen A
We conducted a study to compare the prevalence and antimicrobial resistance profile of Campylobacter isolated from 34 farm-slaughter pair cohorts of pigs raised in conventional and antimicrobial-free (ABF) production systems. Isolates originated from four different states of two geographic regions (region 1--Ohio and Michigan; region 2--Wisconsin and Iowa). A total of 838 fecal and 1173 carcass samples were examined. Campylobacter isolates were speciated using multiplex polymerase chain reaction targeting ceuE and hipO genes. The minimum inhibitory concentration was determined using agar dilution to a panel of six antimicrobials: chloramphenicol, erythromycin, gentamicin, ciprofloxacin, nalidixic acid, and tetracycline. Campylobacter spp. was isolated from 472 of 838 pigs (56.3%). Campylobacter prevalence did not vary significantly based on production system (conventional [58.9%] and ABF [53.7%], odds ratio [OR] 1.4, 95% confidence interval [CI] 0.8-2.6, p = 0.24) or geographic region (region 1 [54.1%] and region 2 [58.2%], OR 1.02, 95% CI 0.6-1.9, p = 0.92). At slaughter plant, Campylobacter prevalence varied based on processing stages (19.4% at pre-evisceration, 25.3% at postevisceration, and 3.2% at postchill). Resistance was common to tetracycline (64.5%), erythromycin (47.9%), and nalidixic acid (23.5%). Campylobacter isolates from conventional production systems were more likely to be erythromycin resistant than from ABF (OR 3.2, 95% CI 1.4-7.2, p = 0.01). The proportion of ciprofloxacin-resistant Campylobacter coli isolates were 3.7% and 1.2% from ABF and conventional production systems, respectively. Thirty-seven out of 1257 C. coli (2.9%) were resistant to both erythromycin and ciprofloxacin, drugs of choice for treatment of invasive human campylobacteriosis. The finding of ciprofloxacin resistance, particularly from ABF herds, has significant implications on the potential role of risk factors other than mere antimicrobial use for production
Migura-Garcia, Lourdes; Ramos, Raül; Cerdà-Cuéllar, Marta
Wildlife is a natural reservoir of Salmonella and Campylobacter, the most important human foodborne pathogens worldwide. Free-living birds have the potential to transport, over large distances, such zoonotic bacteria that may harbor antimicrobial resistance traits. On the northeastern Iberian coast, we assessed the role of Yellow-legged Gulls ( Larus michahellis ) as reservoirs of antimicrobial resistance in Salmonella and thermophilic Campylobacter isolates recovered from gulls at three colonies, with varying degrees of dependence on refuse dumps as food sources. Of the 39 Salmonella isolates we tested, 17 were multiresistant (resistance to three antimicrobial families), with eight being Salmonella enterica serovar Typhimurium. Other clinically relevant Salmonella serovars showing multiresistance included Hadar, Bredeney, and Virchow. Relevant Campylobacter antimicrobial resistances were detected among three Campylobacter jejuni isolates, of which all three showed resistance to nalidixic acid, two were resistant to ciprofloxacin, one was resistant to enrofloxacin, and one was resistant to tetracycline. Our results highlight the importance of free-living gulls with opportunistic feeding habits in the dissemination of enteric pathogens resistant to multiple antimicrobial agents of public health concern.
M’ikanatha, Nkuchia M.; Dettinger, Lisa A.; Perry, Amanda; Rogers, Paul; Reynolds, Stanley M.
In 2010, we surveyed 176 clinical laboratories in Pennsylvania regarding stool specimen testing practices for enteropathogens, including Campylobacter spp. Most (96.3%) routinely test for Campylobacter spp. In 17 (15.7%), a stool antigen test is the sole method for diagnosis. We recommend that laboratory practice guidelines for Campylobacter spp. testing be developed. PMID:22377086
Andrzejewska, M; Klawe, J J; Szczepańska, B; Spica, D
The presence of the flaA, cadF, cdtB and iam genes of Campylobacter spp. was determined with the PCR method. The materials to investigate were 56 C. jejuni and 23 C. coli strains isolated from clinical samples (children and domestic animals). It was found that all of the Campylobacter spp. isolates from children with diarrhoea and domestic animals had cadF gene, responsible for adherence. The flaA gene was present in all Campylobacter spp. isolates derived from children and cats. Occurrence of flaA gene was confirmed in 100% of C. jejuni strains obtained from dogs. The high prevalence of the cdtB gene associated with toxin production was observed in this study (100%-Campylobacter spp. isolates obtained from dogs and cats, 97.9%-Campylobacter spp. isolates from children). The isolates showed a wide variation for the presence of iam gene. The lowest prevalence (23.5%) was detected in Campylobacter spp. obtained from dogs. The highest rates of iam detection (91.6%) were revealed in C. coli isolates from children.
Ladrón de Guevara, C; Gonzalez, J; Peña, P
The genus Campylobacter has become increasingly recognised as the cause of various infections. Campylobacter jejuni and C coli cause acute gastroenteritis in man all over the world. C jejuni enteritis can lead to bacteraemia, but its actual incidence remains unknown. Seven cases of bacteraemia caused by C jejuni or C coli are reported, from the blood of seven patients: five immune deficient adults; a newborn baby; and a patient who had had abdominal surgery. Patients who develop diarrhoea as a result of Campylobacter infection are at risk of bacteraemia thereafter. PMID:8132835
Wassenaar, T M
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. PMID:9227862
Antimicrobial Resistance Profiles of Campylobacter spp. Isolated from Broiler Chicken Meat of Estonian, Latvian and Lithuanian Origin at Estonian Retail Level and from Patients with Severe Enteric Infections in Estonia.
Mäesaar, M; Kramarenko, T; Meremäe, K; Sõgel, J; Lillenberg, M; Häkkinen, L; Ivanova, M; Kovalenko, K; Hörman, A; Hänninen, M-L; Roasto, M
The resistance patterns of Campylobacter spp. isolated from retail broiler chicken meat originating either from Estonia, Lithuania or Latvia collected in Estonia were determined. Additionally, in collaboration with the laboratories of several Estonian hospitals, antimicrobial susceptibility patterns were determined for Campylobacter isolates from patients with severe Campylobacter enteric infections. The isolates were identified at the species level by the PCR method. Respectively, 88.8% of the isolates were C. jejuni, and 11.2% were C. coli. In total, 126 Campylobacter isolates of broiler chicken meat and human origin were tested for minimal inhibitory concentrations (MICs) with the broth microdilution VetMIC(TH) method (National Veterinary Institute; Uppsala, Sweden) for a total of six antimicrobials. Resistance to one or more antimicrobials was detected in 62 (63.3%) of Campylobacter broiler chicken meat isolates and in 20 (71.4%) of human-origin isolates. Large proportions of the broiler chicken meat isolates were resistant to ciprofloxacin (60.2%). Multidrug resistance (i.e. to three or more unrelated antimicrobials) was detected in five (5.1%) C. jejuni isolates. Among the human isolates, 20 (71.4%) were resistant to fluoroquinolones, and two (7.1%) C. jejuni isolates exhibited multidrug resistance. The chicken meat isolates of Estonian origin were the most susceptible. However, a high proportion of fluoroquinolone-resistant C. jejuni isolates were found in Latvian and Lithuanian products. The results of this study indicate that the problems caused by the inappropriate use of antimicrobials extend beyond the country in which a food originates; therefore, both domestic and international interventions and agreements are required to implement common policies on antimicrobial usage and to minimize the emergence of Campylobacter drug resistance.
Campylobacter is a foodborne pathogen which has a potential public health concern worldwide. Due to discriminatory problems encountered by conventional isolation of Campylobacter spp. and its genetic similarities, rapid molecular techniques for its genetic characterization are useful. In this study,...
Comparative performance of isolation methods using Preston broth, Bolton broth and their modifications for the detection of Campylobacter spp. from naturally contaminated fresh and frozen raw poultry meat.
Seliwiorstow, T; De Zutter, L; Houf, K; Botteldoorn, N; Baré, J; Van Damme, I
The performance of different isolation methods was evaluated for the detection of Campylobacter from naturally contaminated raw poultry meat. Therefore, fresh and frozen poultry meat samples were analysed using the standard procedure (ISO 10272-1:2006), enrichment in Preston broth, and enrichment in modified Bolton broth (supplemented with (i) potassium clavulanate (C-BB), (ii) triclosan (T-BB), (iii) polymyxin B (P-BB)). The enrichment cultures were streaked onto both modified charcoal cefoperazone deoxycholate agar (mCCDA) and RAPID'Campylobacter agar (RCA). Moreover, direct plating on mCCDA and RCA was performed to quantify Campylobacter. In total, 33 out of 59 fresh retail meat samples (55.9%) were Campylobacter positive. For both fresh and frozen poultry meat samples, enrichment in Bolton broth (ISO 10272-1:2006) resulted in a higher number of positive samples than enrichment in Preston broth. Supplementation of Bolton broth with potassium clavulanate (C-BB) and triclosan (T-BB) enhanced the Campylobacter recovery from fresh poultry meat compared to non-supplemented Bolton broth, although the use of C-BB was less applicable than T-BB for Campylobacter recovery from frozen samples. Additionally, the use of RCA resulted in a higher isolation rate compared to mCCDA. The present study demonstrates the impact of culture medium on the recovery of Campylobacter from fresh and frozen naturally contaminated poultry meat samples and can support laboratories in choosing the most appropriate culturing method to detect Campylobacter.
von Altrock, Alexandra; Hamedy, Ahmad; Merle, Roswitha; Waldmann, Karl-Heinz
The objective of the study was to determine the prevalence of Campylobacter spp. on surfaces of slaughtered pig livers. Multilocus sequence typing (MLST) was performed to determine the sequence types (STs) of selected Campylobacter coli isolates. Additionally, C. coli and Campylobacter jejuni isolates were tested for antimicrobial susceptibility by the broth dilution method. The minimal inhibitory concentrations were determined for erythromycin, gentamicin, ampicillin, ampicillin/sulbactam, nalidixic acid, ciprofloxacin, tetracycline and trimethoprim/sulphamethoxazole. Samples were taken during the slaughtering process in a slaughterhouse in Lower Saxony, Germany. Altogether, 10% of 1500 surfaces of pig livers from 50 fattening herds was found to be Campylobacter positive, with C. coli as the predominant species (76%) followed by C. jejuni (21%). Resistance to erythromycin and tetracycline was higher in C. jejuni compared to C. coli, whereas C. coli were more resistant to quinolone compared to C. jejuni. Fluoroquinolone resistance is usually associated with cross-resistance to quinolone, but in the presented investigation C. coli as well as C. jejuni showed a higher resistance to ciprofloxacin (28.6% and 20.0%, respectively) than to nalidixic acid (9.5% and 0%, respectively). A high genetic diversity of the C. coli isolates was demonstrated by MLST. Differences in STs and antimicrobial resistance pattern indicate that the Campylobacter strains originated from the pig itself and not from the slaughterhouse. A comparison of the STs with those reported in the C. jejuni/coli PubMLST database showed an overlap of porcine and human isolates, indicating that C. coli isolates from pigs should be considered as potential sources of human infection.
Gilpin, Brent; Robson, Beth; Lin, Susan; Scholes, Paula; On, Stephen
Pulsed-field gel electrophoresis (PFGE) analysis demonstrated that while 76% of patients had only one genotype of campylobacter, 10% carried two different but related genotypes (Dice coefficients > 0.78), and 14% carried at least two unrelated genotypes (Dice coefficients < 0.65). This supports the clustering of Campylobacter isolates with similar PFGE patterns, highlights the need to analyze multiple isolates from both sources and patients, and confirms that caution should be exercised before epidemiological links between patients or sources are dismissed.
Campylobacter spp. are among the most common cause of bacterial foodborne diarrheal illness; poultry has been linked as a primary source of contamination. Detection and enumeration of low numbers of naturally occurring Campylobacter spp. on poultry is difficult due to the presence of competing micro...
Denis, M; M Tanguy; Chidaine, B; Laisney, M-J; Mégraud, F; Fravalo, P
Presence or absence of Campylobacter spp. in water of five rivers upstream from an intake point for drinking water production was investigated, and isolates genetically compared with human, pig and poultry isolates in order to determine their source. River water and drinking water obtained from these rivers were sampled one time per month, over a period of one year, and tested for Campylobacter. Isolates were typed by PFGE. Campylobacter was not detected in treated drinking water, but 50% of the river samples were contaminated. Contamination was observed on the four seasons. In total, 297 Campylobacter isolates were collected and generated 46 PFGE profiles. Campylobacter jejuni was the most frequently detected species in samples (74.1% of the isolates), followed by Campylobacter coli (17.8%) and Campylobacter lari (8.1%). Forty-two of the 46 PFGE profiles were unique. Only one genotype was detected three times in a river during the year and four genotypes in two different rivers. When compared to animal and human Campylobacter PFGE profiles, 14, 11 and one Campylobacter genotypes from water were genetically closed to human, poultry, and pig Campylobacter genotypes, respectively. The Campylobacter population displayed a high level of genetic diversity, suggesting that contamination originated from various origins. Human, poultry and pig were sources of contamination of the river by Campylobacter. Finally, no Campylobacter were detected in drinking water, indicating that the risk of outbreaks due to consumption of drinking water is low.
Chokboonmongkol, C; Patchanee, P; Gölz, G; Zessin, K-H; Alter, T
This study was conducted to determine the prevalence of Campylobacter spp. in broiler flocks by testing cecal contents at slaughter and to detect and quantify Campylobacter on broiler carcass skin samples of the corresponding slaughter batches, to determine antimicrobial resistance patterns of the Campylobacter isolates, and to genotype selected Campylobacter jejuni isolates using multilocus sequence typing analysis. Ninety-eight broiler flocks were included in the study. Intact ceca were randomly taken at the time of evisceration throughout a slaughter batch to detect Campylobacter spp. at the broiler flock level and one whole carcass per slaughter batch was taken for the detection of Campylobacter spp. on broiler skin. The prevalences of Campylobacter spp. in broiler ceca and broiler skin samples were 11.2% (11/98) and 51% (50/98), respectively. Even though most Campylobacter-positive broiler skin samples were contaminated with only up to 230 most probable number per gram, a substantial share (13.3%) showed very high Campylobacter numbers on the broiler skin samples (most probable number = ∞; lower confidence limit T(0) 580/g). From 32 C. jejuni and Campylobacter coli isolates tested, the highest antimicrobial resistance rates were found for ciprofloxacin (81.2%), followed by tetracycline (40.6%), ampicillin (31.2%), and erythromycin (9.4%). All tested strains were sensitive to gentamicin. By multilocus sequence typing analysis, a total of 9 different sequence types were identified among 16 C. jejuni isolates. Campylobacter jejuni isolated from cecal content and carcass skin of the same farm or slaughter batch showed corresponding allelic profiles. Our data suggest that intense cross-contamination during the slaughter process led to a strong increase of Campylobacter prevalence on broiler skin compared with the prevalence in broiler ceca. To reduce Campylobacter prevalences on broiler skin, on-farm biosecurity measures need to be accompanied by control measures
Sánchez, R; Fernández-Baca, V; Díaz, M D; Muñoz, P; Rodríguez-Créixems, M; Bouza, E
Erythromycin, new macrolides, and quinolones are alternatives for the treatment of Campylobacter infections. Concerns related to the emergence of resistance to both groups of drugs have been raised. We studied the evolution of antimicrobial susceptibilities of 275 clinical isolates of microorganisms of the genus Campylobacter isolated in our institution during a 5-year period (1988 to 1992). The microorganisms studied were C. jejuni (n = 230), C. coli (n = 42), and C. fetus (n = 3). The overall resistance rates (determined by the agar dilution method and the recommendations of the National Committee for Clinical Laboratory Standards) were as follows: erythromycin, 2.3%; clarithromycin, 2.3%; azithromycin, 1.9%; ciprofloxacin, 28.5%; norfloxacin, 31%; ofloxacin, 26.3%; and nalidixic acid, 36.8%. The evolution of resistance (percent resistance in 1988 versus percent resistance in 1992) was as follows: erythromycin, 2.6 versus 3.1; clarithromycin, 2.6 versus 3.1; azithromycin, 2.6 versus 3.1; ciprofloxacin, 0 versus 49.5; norfloxacin, 2.6 versus 55.5; ofloxacin, 0 versus 45.6; nalidixic acid, 2.6 versus 56.8. Our data show stable macrolide activity against Campylobacter spp. and the rapid development of quinolone resistance over the last 5 years.
Sison, F B; Chaisowwong, W; Alter, T; Tiwananthagorn, S; Pichpol, D; Lampang, K N; Baumann, M P O; Gölz, G
This study was performed to determine the prevalence and to semiquantify Campylobacter spp. on chicken meat samples at 4 selected local wet markets in Nueva Ecija, Philippines, and to determine the antimicrobial resistance patterns of the Campylobacter isolates. Out of 120 chicken meat samples, 57 (47.5%) were Campylobacter spp. positive. The majority of isolated Campylobacter strains were identified as Campylobacter coli (54.4%) and 45.6% as Campylobacter jejuni. Most of these positive samples (52.6%) showed a very high quantitative Campylobacter contamination (most probable number > 2,400/g, lower confidence limit 580/g). For antimicrobial resistance testing, 44 C. coli/jejuni isolates were tested using the agar disk diffusion method. Out of these, 77.3% were resistant to ampicillin, followed by ciprofloxacin (70.4%), tetracycline (54.6%), erythromycin (20.2%), and gentamicin (11.4%). Of the isolates, 36.4% (n = 16) were resistant to 1 antimicrobial agent, 34.1% (n = 15) were resistance to 3 antimicrobial agents, 13.6% (n = 6) to 2 antimicrobial agents, 9.1% (n = 4) to 4 antimicrobial agents, and 6.8% (n = 3) to all 5 antimicrobial agents tested. Our data demonstrate a high contamination of fresh chicken meat with Campylobacter spp. at retail in the Philippines. The detected high Campylobacter prevalences and quantitative loads on chicken meat at retail in the Philippines highlight the need to implement efficient intervention measures along the food chain and to encourage sanitary handling of poultry meat.
Giacomelli, M; Follador, N; Coppola, L M; Martini, M; Piccirillo, A
Campylobacteriosis is among the most common bacterial causes of human gastroenteritis worldwide and pet ownership has been identified as a risk factor for Campylobacter infection in humans. Since canine and feline prevalence data are scarce in Italy, the present study was carried out to assess the prevalence, species distribution and risk factors for Campylobacter infection in dogs and cats under different husbandry conditions. Rectal swabs were collected from 171 dogs (household pets, n = 100; shelter-housed dogs, n = 50; dogs from breeding kennels, n = 21) and 102 cats (household pets, n = 52; shelter-housed cats, n = 21; free-roaming cats n = 29) in Northern Italy. Campylobacter was isolated from 17% (n = 29) of dogs and 14.7% (n = 15) of cats. C. jejuni was the most common isolate in both species (Campylobacter spp.-positive dogs, 55.2%; Campylobacter spp.-positive cats, 53.3%), followed by C. upsaliensis (Campylobacter spp.-positive dogs, 27.6%; Campylobacter spp.-positive cats, 40%). Other Campylobacter species were rarely detected, but included C. hyointestinalis subsp. hyointestinalis, C. lari and C. coli in dogs and C. coli and C. helveticus in cats. Among considered variables (sex, age, origin, diarrhoea, season of sampling), origin was identified as a risk factor for dogs, with shelter-housed dogs at higher risk than household dogs (odds ratio, 2.84; 95% CI 1.17, 6.92; P = 0.021). The results of this study, particularly the high prevalence of C. jejuni in Campylobacter-positive animals, demonstrated that household and stray dogs and cats in Northern Italy might pose a zoonotic risk for humans. Moreover, biosecurity measures should be improved in dog shelters.
Kashoma, Isaac P.; Kumar, Anand; Sanad, Yasser M.; Gebreyes, Wondwossen; Kazwala, Rudovick R.; Garabed, Rebecca
Abstract Poultry are recognized as a main reservoir of Campylobacter spp. However, longitudinal studies investigating the persistence of Campylobacter on commercial meat turkeys are rare. The objectives of this study were to determine the prevalence, antimicrobial susceptibility, and persistence of genotypically related strains of Campylobacter spp. recovered from three commercial turkey farms in Ohio belonging to a single producer. Eight hundred ten samples were collected from birds aged 1 week to slaughter, consisting of 750 fecal droppings and 60 ceca at slaughter. Overall Campylobacter prevalence was 55.9%. Multiplex polymerase chain reaction (PCR) confirmed 72.3% of all isolates as C. coli, 5.3% as C. jejuni, 10.6% as both, and 11.9% as other Campylobacter spp. PCR restriction fragment length polymorphism of the flaA gene subtyping detected 70 types—62 for C. coli and 8 for C. jejuni isolates—with most (80%) of flaA-types constituting farm homogeneous groups. Multilocus sequence typing of 99 selected Campylobacter isolates resulted in 23 sequence types (STs), consisting of 8 STs for C. jejuni and 15 STs for C. coli isolates. Six novel STs—four for C. jejuni and two—for C. coli, were detected. In a subset of isolates (n=98) tested for antimicrobial resistance, the most common resistance was to tetracycline (95%), followed by azithromycin (43%), while 42% and 18% of the isolates were resistant to ciprofloxacin and erythromycin, respectively. All isolates were susceptible to florfenicol. C. coli isolates displayed a higher proportion of resistance than C. jejuni to most antimicrobials. This study highlights the high prevalence, genotypic diversity, and antimicrobial resistance of Campylobacter spp. in commercial turkey from farm to slaughter. PMID:25184688
Arvanitidou, M; Stathopoulos, G A; Constantinidis, T C; Katsouyannopoulos, V
In order to assess Salmonella, Campylobacter and Yersinia spp. occurrence in surface waters and to compare it with the standard faecal indicator bacteria, 86 river and lake samples, from eight sampling sites in Northern Greece were examined for the presence of these pathogens in parallel to total and faecal coliforms and faecal streptococci. A total of 17 Salmonellae, 14 Campylobacters and 9 Yersiniae were isolated. Only in Salmonella positive samples the geometric means of total and faecal coliforms were found significantly higher (p < 0.01) than in the negative samples, whereas the presence of Campylobacters and Yersiniae may not be predicted by the standard indicator bacteria.
Lin, S M; Molan, P C; Cursons, R T
We report the antimicrobial effect of manuka honey against Campylobacter spp. isolated by a diagnostic laboratory from specimens from a community in New Zealand. The isolates were differentiated according to species level using multiplex PCR. C. jejuni (20 strains) and C. coli (7 strains) were identified. The clinical isolates identified and type culture collection strains of these species were subjected to testing to determine the minimum inhibitory concentration (MIC) of manuka honey using a microdilution technique. The MIC of the manuka honey against all of the Campylobacter tested was found to be around 1% (v/v) honey. The low MIC values suggest that honey might still inhibit the growth of campylobacteria after dilution by fluid in the gut, but the actual concentration of honey that can be achieved in the intestine is unknown. Therefore, clinical investigation is required to establish the efficacy of honey against Campylobacter spp. in the gut environment.
Lay, Kruy Sun; Vuthy, Yith; Song, Ping; Phol, Khem; Sarthou, Jean Louis
Salmonella and Campylobacter are common bacterial pathogens associated with human gastro-enteritis; and raw poultry is considered to be an important source of these bacteria. To evaluate whether the Salmonella serovars and Campylobacter spp. bacteria could be monitored for the purpose of microbial presence, enumeration and antimicrobial resistance in raw poultry, 152 poultry carcasses were randomly selected from 10 markets in retail outlets of Phnom Penh during March 2006 to February 2007. The majority of poultry samples was contaminated by Salmonella serovars (88.2%) and Campylobacter spp. (80.9%). A very high contamination of Salmonella was found at 3-4 log₁₀ CFU/g for 22.4% of samples and of Campylobacter at 7-8 log₁₀ CFU/g for 1.3% of samples. Fifty nine different Salmonella serovars contaminated 134 poultry carcasses; five most prevalent serovars covered 29.1% of serovars isolates (Anatum, Typhimurium, Corvallis, Stanley and Enteritidis). Three Campylobacter species contaminating 123 raw poultry were Campylobacter jejuni (50.0%), Campylobacter coli (29.0%) and Campylobacter lari (21.0%). High antibiotic resistance percentages were found among Salmonella serovars and Campylobacter spp. isolates. This study revealed that raw poultry at the retail outlets in Phnom Penh markets are contaminated with high prevalences of food-borne pathogens, and communicating the importance of minimizing this risk in reducing human infections.
Li, Beibei; Ma, Licai; Li, Yingli; Jia, Haiyan; Wei, Jianchao; Shao, Donghua; Liu, Ke; Shi, Yuanyuan; Qiu, Yafeng; Ma, Zhiyong
This study was conducted to determine the prevalence of antimicrobial resistance in Campylobacter spp. isolates from broilers in live bird markets (LBMs). A total of 209 Campylobacter spp. isolates (84 Campylobacter jejuni; 125 Campylobacter coli) were recovered from 364 broiler cecum samples collected from five LBMs in Shanghai, China. Minimum inhibitory concentrations of 13 antimicrobials were determined using agar dilution method. More than 96% of the Campylobacter spp. isolates were resistant to quinolones and tetracyclines. A high prevalence of macrolide resistance (erythromycin, 84.0%; azithromycin, 80.8%) was observed in C. coli, but not in C. jejuni (erythromycin, 6.0%; azithromycin, 2.4%). C. coli also showed significantly higher resistance than C. jejuni to clindamycin, gentamicin, and kanamycin. In contrast, C. coli isolates had lower resistance to florfenicol than the C. jejuni isolates. The majority of the C. jejuni (88.1%) and C. coli (97.6%) isolates exhibited multidrug resistance (MDR) to three or more classes of antimicrobials. All of the 208 ciprofloxacin-resistant Campylobacter spp. isolates were positive for the C257T mutation of the gyrA gene. In addition, the tet(O) gene was identified in all of the 202 doxycycline-resistant Campylobacter spp. isolates. Furthermore, 75.7% and 20.4% of the 103 azithromycin-resistant Campylobacter spp. isolates were positive for the A2075G mutation of the 23S rRNA gene and the presence of the erm(B) gene, respectively. Moreover, the cat gene was found in 14.3% (8/56) and 76.8% (73/95) of the chloramphenicol-resistant C. jejuni and C. coli isolates, respectively. To the best of our knowledge, this is the first report of the prevalence of antimicrobial resistance among Campylobacter spp. isolates originating from LBMs. The high prevalence of MDR Campylobacter spp. isolates in LBMs highlights the need to implement efficient intervention measures to control not only Campylobacter contamination in LBMs but also
Rhynd, Kamara J R; Leighton, Patrick A; Elcock, David A; Whitehall, Pamela J; Rycroft, Andrew; Macgregor, Shaheed K
From April to July 2005, rectal swabs were collected from 48 free-ranging small Asian mongooses (Herpestes javanicus) on the east and south coasts of Barbados and analyzed for Salmonella and Campylobacter spp. Salmonella was recovered in 21.12% (7/33) of mongooses at the east-coast site and 26.67% (4/15) at the south-coast site. Four serotypes were isolated: Salmonella enterica serovar Rubislaw, Kentucky, Javiana, and Panama. One east-coast sample of 11 tested for Campylobacter was positive (9.09%). These results indicate that mongooses in Barbados are carriers and shedders of Salmonella and Campylobacter spp. and are a potential wildlife reservoir for these enteropathogens.
Griggs, Deborah J.; Johnson, Maggie M.; Frost, Jennifer A.; Humphrey, Tom; Jørgensen, Frieda; Piddock, Laura J. V.
Five commercial broiler flocks were treated with a fluoroquinolone for a clinically relevant infection. Fresh feces from individual chickens and environmental samples were cultured for campylobacters before, during, and weekly posttreatment until slaughter. Both Campylobacter jejuni and C. coli were isolated during all treatment phases. An increased proportion of quinolone-resistant strains was seen during treatment, and these strains persisted posttreatment. One quinolone-resistant isolate of each species, each serotype, and each phage type from each sample at all treatment phases was examined for its phenotype and mechanism of resistance. Two resistant phenotypes were isolated: Nalr Cipr and Nalr Cips. The majority (269 of 290) of fluoroquinolone-resistant isolates, whether they were C. jejuni or C. coli, had a mutation in gyrA that resulted in the substitution Thr-86→Ile. The other gyrA mutations detected were Thr-86→Ala (n = 17) and Asp-90→Asn (n = 10). The genotypic variation, based on the silent mutations in gyrA identified by the denaturing high-performance liquid chromatography pattern and DNA sequencing, was used to supplement typing data and provided evidence for both the spread of preexisting resistant strains and the selection of spontaneous resistant mutants in treated flocks. Multidrug resistance was significantly (P < 0.01) associated with resistance to ciprofloxacin. Twenty-five percent (73 of 290) of ciprofloxacin-resistant isolates but only 13% (24 of 179) of susceptible isolates were resistant to three or more unrelated antimicrobial agents. In conclusion, quinolone-resistant campylobacters were isolated from commercial chicken flocks in high numbers following therapy with a veterinary fluoroquinolone. Most ciprofloxacin-resistant isolates had the GyrA substitution Thr-86→Ile. Resistant isolates were isolated from the feces of some flocks up to the point of slaughter, which may have consequences for public health. PMID:15673754
Dudzic, A; Urban-Chmiel, R; Stępień-Pyśniak, D; Dec, M; Puchalski, A; Wernicki, A
1. The aim of this study was to evaluate the occurrence of Campylobacter spp. in domestic and free-living pigeons and to evaluate the antibiotic resistance profiles. 2. The material consisted of cloacal swabs obtained from 108 homing pigeons and fresh faeces from 72 wild birds from Lublin and its vicinity. The identification of strains isolated on differential/selective media for Campylobacter spp. was carried out by MALDI-TOF and PCR. The susceptibility to antibiotics was evaluated by minimum inhibitory concentration (MIC) in Mueller-Hinton broth. 3. A total of 35 strains of Campylobacter spp. were isolated; 27 were identified as Campylobacter jejuni and 8 as Campylobacter coli. Over half of the isolates were resistant to erythromycin and streptomycin, 40% of strains were resistant to tetracycline and ampicillin and 37% isolates were resistant to amoxicillin. Resistance to two or more antibiotics was observed in all strains tested. 4. The results indicate that both domestic and free-living pigeons are reservoirs for bacteria of the genus Campylobacter, which are characterised by varied and growing resistance to commonly used antibiotics.
Rahimi, E; Momtaz, H; Ameri, M; Ghasemian-Safaei, H; Ali-Kasemi, M
The objective of this study was to determine the prevalence and antimicrobial resistance of Campylobacter spp. isolated from chicken carcasses during different stages of broiler processing in a major commercial poultry processing plant in southwestern Iran. Overall, 84 chicken carcasses were sampled from 4 sites along the processing line during a total of 7 visits. In addition, 14 water samples from the chiller tank were taken. Using the cultural method, 186 of 336 (55.4%) carcasses were positive for Campylobacter. Campylobacter jejuni was more frequently isolated (89.4%) than Campylobacter coli (10.6%). The frequency of Campylobacter spp. on carcasses was 54.8% after defeathering, 51.2% after evisceration, 69.0% 20 min after the chilling period started, and 46.4% 24 h after the chilling period completed. Campylobacter was positive in 85.7% of the samples taken from the chilling water. The frequency of Campylobacter spp.-positive carcasses was reduced in complete chilled chickens but not during the slaughtering process. Susceptibilities of Campylobacter isolates were determined for 10 antimicrobial drugs using the disk diffusion method. Of the 198 Campylobacter isolates tested, 92.9% were resistant to one or more antimicrobial agents. Resistance to tetracycline was the most common finding (78.3%), followed by resistance to ciprofloxacin (62.1%), nalidixic acid (58.6%), and enrofloxacin (44.4%).
O'Sullivan, N; Benjamin, J; Skirrow, M B
Isolates of Campylobacter jejuni, C coli, C fetus and C laridis were tested for agglutination reactions with a panel of five lectins: Arachis hypogaea, Bauhinia purpurea, Solanum tuberosum, Triticum vulgaris and Wisteria floribunda. Twenty three patterns of agglutination (lectin types) were recorded among 376 isolates. Patterns were consistent and reproducible. Only 4.5% of isolates were untypable because of autoagglutination. Some lectin types were found exclusively or predominantly in a species, but others were shared between species. Forty two per cent of C jejuni and 35% of C coli isolates belonged to lectin type 4. There was no apparent correlation between lectin type and serotype; different lectin types were found among strains of single Penner and Lior serotypes. Lectin typing is a simple and economical procedure suitable for use in non-specialist laboratories, either as an adjunct to serogrouping or, after further development, as a sole typing scheme. PMID:2262570
Han, Feifei; Lestari, Shofiyah Ika; Pu, Shuaihua; Ge, Beilei
Effective in September 2005, enrofloxacin, a fluoroquinolone group antimicrobial, was withdrawn from use in the U.S. poultry farms. In this 1-year study initiated in October 2006, we isolated and characterized Campylobacter spp. from Louisiana retail conventionally raised (n = 141) and organic (n = 53) chickens as a comparison to evaluate the postban bacterial resistance to antimicrobials. Campylobacter was present in 43.3% of the chickens; similar rates were observed among conventional and organic chickens. A total of 165 Campylobacter isolates were recovered, with Campylobacter jejuni being the predominant species (66.7%). No apparent seasonal trend was deduced from the prevalence data. Further, the two main conventional and one organic chicken brands did not carry significantly different rates of Campylobacter (p > 0.05). The most common resistance observed was to tetracycline (31.5%), followed by erythromycin (20%) and ciprofloxacin (6.1%). No resistance to gentamicin was identified. All Campylobacter isolates recovered from organic chickens (n = 48) were susceptible to ciprofloxacin, compared to 8.5% resistance rate for those from conventional chickens (n = 117). Additionally, the resistance rate to erythromycin was significantly higher in Campylobacter isolates from conventional chickens (23.9%) than those from organic chickens (10.4%; p < 0.05). Our results demonstrated a low prevalence and low ciprofloxacin resistance rate of Campylobacter in Louisiana retail chickens after the enrofloxacin ban. Further studies involving a larger sample size over time are warranted to better assess the effects of banning enrofloxacin use in poultry and the levels of fluoroquinolone-resistant Campylobacter.
During samplings of reptiles for Epsilonproteobacteria, Campylobacter strains were isolated from lizards and chelonians not belonging to any of the established taxa. Initial AFLP, PCR, and 16S rRNA sequence analysis showed that these strains were most closely related to Campylobacter fetus and Campy...
Rahimi, Ebrahim; Ameri, Mehrdad; Kazemeini, Hamid Reza
Campylobacter spp. are one of the most common causes of acute bacterial gastroenteritis in human beings which are transmitted mostly via food originating from animals. This study was conducted to determine the prevalence and antimicrobial resistance of Campylobacter spp. isolated from retail raw meats in Iran. From June 2008 to June 2009, a total of 722 raw meat samples from camel (n = 107), beef (n = 190), lamb (n = 225), and goat (n = 180) were purchased from randomly selected retail outlets in Isfahan and Yazd, Iran, and were evaluated for the presence of Campylobacter spp. In this study, 50 of the 722 meat samples (6.9%) were contaminated with Campylobacter spp. The highest prevalence of Campylobacter spp. was found in lamb meat (12.0%), followed by goat meat (9.4%), beef meat (2.4%), and camel meat (0.9%). The most prevalent Campylobacter spp. isolated from the meat samples was Campylobacter jejuni (84.0%); the remaining isolates were Campylobacter coli (16.0%). Susceptibilities of 50 Campylobacter isolates were determined for 10 antimicrobial drugs using the disk-diffusion assay. Resistance to tetracycline was the most common finding (68.0%), followed by resistance to ciprofloxacin (46.0%) and nalidixic acid (40.0%). All of the isolates were susceptible to erythromycin, gentamicin, and chloramphenicol. Significantly higher prevalence rates of Campylobacter spp. (p < 0.05) were found in the lamb meat samples taken in spring (20.0%) and summer (18.9%). To our knowledge, this study is the first report of the isolation of Campylobacter spp. from raw camel, lamb, and goat meat in Iran.
Stoddard, Robyn A; Gulland, M D Frances; Atwill, E Rob; Lawrence, Judy; Jang, Spencer; Conrad, Patricia A
Campylobacter and Salmonella spp. prevalence and antimicrobial drug sensitivity were determined in northern elephant seals that had not entered the water and seals that were stranded on the California coast. Stranded seals had a higher prevalence of pathogenic bacteria, possibly from terrestrial sources, which were more likely to be resistant.
Korsak, Dorota; Maćkiw, Elżbieta; Rożynek, Elżbieta; Żyłowska, Monika
The purpose of the present study was to determine the prevalence of thermophilic Campylobacter in poultry, pork, and beef meat at the retail level and to identify the main categories of meat representing the most significant reservoirs of Campylobacter. A monitoring study was conducted throughout Poland from 2009 to 2013. A total of 1,700 fresh meat samples were collected from supermarkets, large retail outlets, and smaller stores. Thermophilic Campylobacter species were detected in 690 (49.3%) of 1,400 poultry samples collected from retail trade. Strains were isolated from 50.2 and 41.1% of raw chicken and turkey meat samples, respectively, and from 50.1 and 42.6% of raw chicken and turkey giblets. The incidence of Campylobacter spp. on pork (10.6%) and beef (10.1%) was significantly lower than on poultry. Campylobacter jejuni was the most prevalent Campylobacter species in chicken (46.6%), pork (68.6%), and beef (66.7%), and Campylobacter coli was the most frequently isolated Campylobacter species in turkey meat (71.2%). This study revealed that retail raw meats are often contaminated with Campylobacter; however, the prevalence of these pathogens is markedly different in different meats. Raw retail meats are potential vehicles for transmitting foodborne diseases, and our findings stress the need for increased implementation of hazard analysis critical control point programs and consumer food safety education efforts.
Gilbert, Maarten J; Kik, Marja; Miller, William G; Duim, Birgitta; Wagenaar, Jaap A
During sampling of reptiles for members of the class Epsilonproteobacteria, strains representing a member of the genus Campylobacter not belonging to any of the established taxa were isolated from lizards and chelonians. Initial amplified fragment length polymorphism, PCR and 16S rRNA sequence analysis showed that these strains were most closely related to Campylobacter fetus and Campylobacter hyointestinalis. A polyphasic study was undertaken to determine the taxonomic position of five strains. The strains were characterized by 16S rRNA and atpA sequence analysis, matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectrometry and conventional phenotypic testing. Whole-genome sequences were determined for strains 1485E(T) and 2463D, and the average nucleotide and amino acid identities were determined for these strains. The strains formed a robust phylogenetic clade, divergent from all other species of the genus Campylobacter. In contrast to most currently known members of the genus Campylobacter, the strains showed growth at ambient temperatures, which might be an adaptation to their reptilian hosts. The results of this study clearly show that these strains isolated from reptiles represent a novel species within the genus Campylobacter, for which the name Campylobacter iguaniorum sp. nov. is proposed. The type strain is 1485E(T) ( = LMG 28143(T) = CCUG 66346(T)).
Hald, B; Madsen, M
Living in a household with a dog or cat has previously been identified as a significant risk factor for acquiring campylobacteriosis, in particular, with reference to Campylobacter upsaliensis infection. In a cross-sectional study carried out in Denmark between August and December 1996, 72 healthy puppies and 42 healthy kittens, aged between 11 and 17 weeks, were sampled for fecal campylobacter shedding by culture of rectal swab specimens on blood-free agar base with cefoperazone at 32 mg/liter and amphotericin at 10 mg/liter and on blood-free agar base with cefoperazone at 8 mg/liter, teicoplanin at 4 mg/liter, and amphotericin at 10 mg/liter. Additionally, with respect to the C. upsaliensis transmission potential of poultry, a chicken cloacal swab sample from each of 100 different broiler flocks was included in the study for comparison. We found 21 (29%) of the puppies positive for Campylobacter spp., with a species distribution of 76% C. jejuni, 5% C. coli, and 19% C. upsaliensis. Of the kittens examined, two (5%) excreted campylobacters; both strains were C. upsaliensis. None of the chicken samples examined were found to be positive for C. upsaliensis. We concluded that young puppies and kittens are potential transmitters of human-pathogenic Campylobacter spp., including C. upsaliensis, while poultry seems negligible in C. upsaliensis epidemiology. PMID:9399557
Jacobs-Reitsma, W F; van de Giessen, A W; Bolder, N M; Mulder, R W
Broiler flocks on two Dutch poultry farms were screened weekly for the presence of campylobacter in fresh caecal droppings during eight consecutive production cycles. Hatchery and fresh litter samples were taken at the start of each new cycle. Water, feed, insects, and faeces of domestic animals, present on the farms were also included in the sampling. Penner serotyping of isolates was used to identify epidemiological factors that contribute to campylobacter colonization in the broiler flocks. Generally, broiler flocks became colonized with campylobacter at about 3-4 weeks of age with isolation percentages of 100%, and stayed colonized up to slaughter. A similar pattern of serotypes was found within the various broiler houses on one farm during one production cycle. New flocks generally showed also a new pattern of serotypes. Most serotypes isolated from the laying hens, pigs, sheep and cattle were different from those isolated from the broilers at the same time. Campylobacter serotypes from darkling beetles inside the broiler houses were identical to the ones isolated from the broilers. No campylobacter was isolated from any of the hatchery, water, feed or fresh litter samples. Conclusive evidence of transmission routes was not found, but results certainly point towards horizontal transmission from the environment. Horizontal transmission from one broiler flock to the next one via a persistent contamination within the broiler house, as well as vertical transmission from breeder flocks via the hatchery to progeny, did not seem to be very likely.
Schroeder, Matthew W; Eifert, Joseph D; Ponder, Monica A; Schmale, David G
Campylobacter spp. have been isolated from live poultry, production environments, processing facilities, and raw poultry products. Environmental sampling in a poultry grow-out house, combined with carcass rinse sampling from the same flock, may provide a relative relationship between pre- and postharvest Campylobacter contamination. Air samples, fecal/litter samples, and feed/drink line samples were collected from 4 commercial chicken grow-out houses in western Virginia between September 2011 and January 2012. Birds from each sampled house were the first flock slaughtered the following day and were then sampled by postchill carcass rinses. Campylobacter, from postenrichment samples, was detected in 27% (32/120) of house environmental samples and 37.5% (45/120) of carcass rinse samples. All environmental sample types from each house included at least one positive sample except the house 2 air samples. The sponge sample method was found to have a significantly higher (P < 0.05) proportion of Campylobacter-positive samples (45%) than the fecal/litter samples (20%) and air samples (15%) when sample types of all the houses were compared. The proportion positive for the fecal/litter samples postenrichment, for each flock, had the highest correlation (0.85) to the proportion of positive carcass rinse samples for each flock. Environmental samples from house 1 and associated carcass rinses accounted for the largest number of Campylobacter positives (29/60). The fewest number of Campylobacter positives, based on both house environmental (4/30) and carcass rinse samples (8/30), was detected from flock B. The results of this study suggest that environmental sampling in a poultry grow-out house, combined with carcass rinse sampling from the same flock, have the potential to provide an indication of Campylobacter contamination and transmission. Campylobacter qualitative levels from house and processing plant samples may enable the scheduled processing of flocks with lower
Stella, Simone; Soncini, Gabriella; Ziino, Graziella; Panebianco, Antonio; Pedonese, Francesca; Nuvoloni, Roberta; Di Giannatale, Elisabetta; Colavita, Giampaolo; Alberghini, Leonardo; Giaccone, Valerio
Retail poultry meat is a crucial vehicle for consumers' exposure to Campylobacters, but no official controls are currently applied in Italy. The aim of this study was the evaluation of Campylobacter contamination of a wide range of poultry meats marketed in Italy. N. 472 chicken and turkey meat samples (sectioned meats, offal, meat preparations and products) were taken from slaughterhouses, deboning plants and different retailers and submitted to detection/enumeration of Campylobacter spp. The isolates were identified by phenotypic and biomolecular techniques. Campylobacter spp. was detected in 34.1% of the samples, with general low counts. Higher values were observed in offal (especially liver) and sectioned meats, with significantly higher rates in skin-on samples (86.8% vs 32.7%). Minced meat preparations showed lower prevalence (22.4% vs 58.3%) and counts than whole pieces. Decreasing rates were observed among slaughterhouses (80%), deboning plants (49%), butcher's shops (37%) and large scale retailers (25%). Sectioned chicken meats were significantly more contaminated than turkey meats. Almost all the isolates were identified as C. jejuni or C. coli, with similar prevalences (18.4% and 20.5%, respectively); C. jejuni was predominant only in samples from slaughterhouses/deboning plants. For setting future control programs, meat typology should be considered the main critical factor.
Badlík, Marián; Holoda, Emil; Pistl, Juraj; Koscová, Jana; Sihelská, Zuzana
This work focused on the isolation of thermophilic Campylobacter spp. in samples obtained from dog droppings. There were 135 samples collected and examined from both clinically healthy and diseased dogs from households, clinics, rehabilitation centres and dog shelters in eastern Slovakia. The isolation of the Campylobacter spp. was achieved by the use of combined selective cultivation methods, followed by confirmation and species identification of the isolates using the PCR method.The overall prevalence of Campylobacter in dogs was 30.4%. Statistically significant differences were recorded (P < 0.05) within the age groups of all dogs examined: 40.6% of the older dogs (> or = 1 year) tested positive, compared to 19.7% of the younger ones (< 1 year). There was no significant difference in relation to dog gender. The most frequently isolated species was Campylobacter (C.) jejuni, present in 51.2% of all positive samples. Campylobacter coli was present in 9.8% of the samples. The remaining positive samples (39%) were confirmed as C upsaliensis, based on phenotypic traits. The highest prevalence of Campylobacter was found in samples from shelters (50%) and the lowest in those from households (11.5%), with samples from rehabilitation centres (42.3%) and clinics (18.8%) coming in second and third place.The high prevalence of Campylobacter confirms the hypothesis that dogs, mainly the ones kept in groups, are a source of Campylobacter spp. Further investigation is required to determine to what extent infected dogs may be a potential source of infection in humans.
Khoshbakht, Rahem; Tabatabaei, Mohammad; Hoseinzadeh, Saeid; Raeisi, Mojtaba; Aski, Hesamaddin Shirzad; Berizi, Enayat
Although poultry meat is considered as the main source for human Campylobacter infections, there is limited information about non-poultry sources. The present study was aimed to investigate the prevalence and the antibiotic resistance of thermophilic Campylobacter spp. in fecal samples of the cattle and sheep in Shiraz, Iran. A total of 302 fecal samples were obtained from clinically healthy, slaughtered cattle and sheep from Shiraz slaughterhouse. The animals were clinically healthy before being slaughtered. The samples were cultured according to the specific cultivation method under thermophilic conditions. The susceptibility of Campylobacter isolates were determined for 13 antimicrobial agents. All enriched samples and cultured isolates were targeted for polymerase chain reaction (PCR) detection of 16S rRNA and multiplex PCR for determining their species. Among 302 fecal samples, 65 (21.5%) and 205 (67.8%) samples were positive for the presence of Campylobacter species with the cultivation and PCR techniques, respectively. All 65 distinct isolates were susceptible to neomycin and colistin and the isolates showed high resistance to cephalotin (83.0%) and ciprofloxacin (67.7%). After the multiplex PCR, 78.5% of total positive samples showed the simultaneous presence of Campylobacter jejuni and Campylobacter coli. In conclusion, the results emphasized that non-poultry farms are important as a possible source of Campylobacter infections. PMID:27872721
Tassew, Haimanot; Asrat, Daniel
Introduction. Campylobacter is one of the leading bacterial causes of food-borne disease. The prevalence of Campylobacter species resistant to antimicrobial agents is increasing. This study is intended to determine prevalence and antimicrobial susceptibility patterns of Campylobacter species among under-five children with diarrhea. Methodology. A cross-sectional study was conducted among 227 under-five children with diarrhea from July to October 2012 at Jimma town. Isolation and identification of Campylobacter species were performed using standard bacteriological techniques. Antimicrobial susceptibility test was performed following standard protocol. Chi-square and Fisher's exact tests were used for analysis. Results. From 227 under-five children, 16.7% were positive for Campylobacter spp.; isolates, C. jejuni, C. coli, and C. lari, accounted for 71.1%, 21.1%, and 7.9%, respectively. Higher rate of resistance was observed to ampicillin 76.3%, trimethoprim-sulfamethoxazole (68.4%), tetracycline (39.5%), chloramphenicol (31.6%), clindamycin (26.3%), and doxycycline (23.7%). Erythromycin, ciprofloxacin, gentamicin, norfloxacin, and nalidixic acid were effective for more than 80% of the isolates. Multiple drug resistance was observed among 78.9% of all the three spp. Conclusions. Isolation rate of Campylobacter spp. was high. C. lari was reported for the first time at this study area. Higher rate of resistance was observed to the commonly used drugs. PMID:26904735
Osaili, Tareq M; Alaboudi, Akram R; Al-Akhras, Rani R
A total of 140 broiler flocks presented for slaughtering at Amman slaughterhouse were tested for Campylobacter spp. via collection of cloacal swabs from live birds, feathered skin samples at prescalding, and skin samples at postscalding (62°C or 57°C scalding temperature), postevisceration, and postchilling. The results indicated that 40% of the flocks tested by cloacal swabs, 34% at prescalding, 32% at post 57°C scalding, and 32% postevisceration were harboring Campylobacter jejuni. None of the skin samples collected from dressed birds at postscalding (62°C) or postwashing-chilling steps (regardless of scalding temperature) revealed the presence of C. jejuni. Thirty eight isolates were tested for susceptibility to ten antimicrobials by using the microbroth dilution method. Almost 50% of the isolates were multidrug resistant to 9 or 10 out of the ten tested antimicrobials. The other half of tested isolates were sensitive to erythromycin, tetracycline, doxycyclin, chlortetracycline, ciprofloxacin, enorfloxacin, gentamycin, tilmicosin, amoxicillin, and trimethoprim.
Medeiros, Diane T; Sattar, Syed A; Farber, Jeffrey M; Carrillo, Catherine D
The occurrence of Campylobacter spp. in a variety of foods from Ottawa, Ontario, Canada, and raw milk samples from across Canada was determined over a 2-year period. The samples consisted of 55 raw foods (chicken, pork, and beef), 126 raw milk samples from raw milk cheese manufacturers, and 135 ready-to-eat foods (meat products, salads, and raw milk cheeses). Campylobacter jejuni was detected in 4 of the 316 samples analyzed: 1 raw beef liver sample and 3 raw chicken samples. An isolation rate of 9.7% was observed among the raw chicken samples tested. This study also investigated the role of cross-contamination in disseminating Campylobacter from raw poultry within a food service operation specializing in poultry dishes. Accordingly, kitchen surfaces within a restaurant in Ottawa, Ontario, were sampled between March and August 2001. Tests of the sampling method indicated that as few as 100 Campylobacter cells could be detected if sampling was done within 45 min of inoculation; however, Campylobacter spp. were not detected in 125 swabs of surfaces within the kitchens of this food service operation. Despite the reported high prevalence of Campylobacter spp. in raw poultry, this organism was not detected on surfaces within a kitchen of a restaurant specializing in poultry dishes.
Jurado-Tarifa, E; Torralbo, A; Borge, C; Cerdà-Cuéllar, M; Ayats, T; Carbonero, A; García-Bocanegra, I
Infections caused by thermotolerant Campylobacter spp. and Salmonella spp. are the leading causes of human gastroenteritis worldwide. Wild birds can act as reservoirs of both pathogens. A survey was carried out to determine the prevalence, genetic diversity and antimicrobial resistance of thermotolerant Campylobacter and Salmonella in waterfowl used as decoys and wild raptors in Andalusia (Southern Spain). The overall prevalence detected for Campylobacter was 5.9% (18/306; CI95%: 3.25-8.52) in decoys and 2.3% (9/387; CI95%: 0.82-3.83) in wild raptors. Isolates were identified as C. jejuni, C. coli and C. lari in both bird groups. Salmonella was isolated in 3.3% (10/306; CI95%: 2.3-4.3) and 4.6% (18/394; CI95%: 3.5-5.6) of the decoys and raptors, respectively. Salmonella Enteritidis and Typhimurium were the most frequently identified serovars, although Salmonella serovars Anatum, Bredeney, London and Mikawasima were also isolated. Pulsed-field gel electrophoresis analysis of isolates showed higher genetic diversity within Campylobacter species compared to Salmonella serovars. Campylobacter isolates showed resistance to gentamicin, ciprofloxacin and tetracycline, while resistance to erythromycin and tetracycline was found in Salmonella isolates. The results indicate that both decoys and raptors can act as natural carriers of Campylobacter and Salmonella in Spain, which may have important implications for public and animal health.
Rossi, M; Hänninen, M L; Revez, J; Hannula, M; Zanoni, R G
In order to study the occurrence and co-infection of different species of Campylobacter, enteric Helicobacter and Anaerobiospirillum in dogs and cats and define a possible association between these microrganisms and gastrointestinal disorders, 190 dogs and 84 cats, either healthy or with diarrhea, were sampled between 2002 and 2003. Thirty-three C. upsaliensis, 17 C. jejuni, 2 C. helveticus, 1 C. lari isolates from dogs and 14 C. helveticus, 7 C. jejuni, 6 C. upsaliensis isolates from cats were identified using species-specific PCR and phenotypic tests. Whole cell protein profile analysis, phenotypic tests, PCR-RFLP of gyrB and a phylogenetic study of partial groEL and 16S rRNA sequences were used to identify 37 H. bilis, 22 H. canis and 14 H. cinaedi in dogs and 12 H. canis, 5 H. bilis and 2 H. cinaedi in cats. Whole cell protein profile analysis, phenotypic tests and species-specific PCR of 16S rRNA were used to identify 14 A. succiniciproducens, 12 A. thomasii isolates and one unidentified Anaerobiospirillum sp. isolate in dogs and 3 A. thomasii isolates in cats. Fifty-two animals (19%) were positive for the isolation of more than one genus. No significant statistical correlation was found between any isolates of Campylobacter, Helicobacter or Anaerobiospirillum spp. or the various co-infection rates, and the presence of diarrhea in either dogs or cats. Campylobacter isolates were also tested for antibiotic resistance using the agar dilution method.
American Society for Microbioloc% Distribution and Polymorphism of the Flagellin Genes from Isolates of Campylobacter coli and Campylobacter jejuni RICHARD...in Campylobacter jejuni . serogroups both the flaA and flaB genes are extremely Mol. M;crobiol. 5:1151-1158. z homologous. Within most LIO heat-labile...irllwn hungatei. J1. Bacteriol. 123:-28 proteins of Campylobacter jejuni 81116. Infect. Immun. 59: 42. Thomashow, L S., and S. C. Rittenberg. 198
Trimoulinard, A; Beral, M; Henry, I; Atiana, L; Porphyre, V; Tessier, C; Leclercq, A; Cardinale, E
One of the most popular meat products of the local "cuisine" is sausage composed with 100% chicken or 100% pork. In this study, we aimed to determine the presence of Salmonella spp., Campylobacter spp. and Listeria spp. in chicken- and pork-sausages, quantify Salmonella spp. population and identify the factors that could be associated with contamination in the outlets. Two hundred and three batches of pork and chicken sausages were randomly collected from 67 local outlets (supermarkets, groceries and butcher shops). Salmonella spp. was detected in 11.8% (95% confidence interval (CI): [10.0; 13.5]) of samples, Campylobacter spp. in 1.5% [0.7; 4.2] and Listeria monocytogenes in 5.9% [4.4; 7.3]. Most probable number of Salmonella spp. varied between 6cfu per gram to 320cfu per gram. Salmonella serotypes isolated from pork and chicken sausages were S. Typhimurium (45.8%), S. London (20.8%), S. Derby (16.7%), S. Newport (8.33%), S. Blockley (4.2%) and S. Weltevreden (4.17%). Using a logistic (mixed-effect) regression model, we found that Salmonella spp. contamination was positively associated with sausages sold in papers or plastic bags and no control of rodents. Chicken sausages were associated with a decreasing risk of Salmonella contamination. Listeria monocytogenes contamination was positively associated with the presence of fresh rodent droppings in the outlet and negatively when the staff was cleaning regularly their hands with soap and water or water only. All the sampled outlets of Reunion Island were not equivalent in terms of food safety measures. Increasing awareness of these traders remains a cornerstone to limit the presence of Salmonella spp. and Listeria spp. in sausages, particularly in a tropical context (high temperature and humidity).
Introduction: Campylobacter spp. are known to be among the most common bacteria to cause diarrheal illness, with poultry being linked as the primary source of contamination related to foodborne illness. Enumeration and detection of low numbers of naturally occurring Campylobacter spp. on poultry pro...
Yee, Emma; Kik, Marja; Wagenaar, Jaap A.; Duim, Birgitta
Campylobacter iguaniorum has been isolated from reptiles. This Campylobacter species is genetically related to Campylobacter fetus and Campylobacter hyointestinalis. Here we present the first whole-genome sequence for this species. PMID:25146144
Gilbert, Maarten J; Miller, William G; Yee, Emma; Kik, Marja; Wagenaar, Jaap A; Duim, Birgitta
Campylobacter iguaniorum has been isolated from reptiles. This Campylobacter species is genetically related to Campylobacter fetus and Campylobacter hyointestinalis. Here we present the first whole-genome sequence for this species.
Wesley, I. V.; Wells, S. J.; Harmon, K. M.; Green, A.; Schroeder-Tucker, L.; Glover, M.; Siddique, I.
Campylobacter jejuni, Campylobacter coli, and Arcobacter spp. were detected in feces of healthy dairy cows by highly specific multiplex-PCR assays. For C. jejuni, at this one-time sampling, cows from 80.6% of farm operations (n = 31) and 37.7% of individual dairy cattle fecal samples (n = 2,085) were positive. Farm management factors were correlated with prevalence in herds in which >25% of cows were positive for C. jejuni. Statistical significance was set at a P of 0.20. Using these criteria, application of manure with broadcast spreaders (P = 0.17), feeding of whole cottonseed or hulls (P = 0.17) or alfalfa (P = 0.15), and accessibility of feed to birds (P = 0.17) were identified as possible risk factors for C. jejuni infection. C. coli was detected in at least one animal in 19.4% of operations and 1.8% of individual cows (n = 2,085). At the herd level, use of broadcaster spreaders was not a risk factor for C. coli infection. For Arcobacter, cows from 71% of dairy operations (n = 31) and 14.3% of individual dairy cattle fecal samples (n = 1,682) were positive. At the herd level, for Arcobacter spp., feeding of alfalfa (P = 0.11) and use of individual waterers (P = 0.19) were protective. This is the first description of Arcobacter spp. in clinically healthy dairy cattle and the first attempt to correlate their presence with C. jejuni. PMID:10788372
Wesley, I V; Wells, S J; Harmon, K M; Green, A; Schroeder-Tucker, L; Glover, M; Siddique, I
Campylobacter jejuni, Campylobacter coli, and Arcobacter spp. were detected in feces of healthy dairy cows by highly specific multiplex-PCR assays. For C. jejuni, at this one-time sampling, cows from 80.6% of farm operations (n = 31) and 37.7% of individual dairy cattle fecal samples (n = 2,085) were positive. Farm management factors were correlated with prevalence in herds in which >25% of cows were positive for C. jejuni. Statistical significance was set at a P of 0.20. Using these criteria, application of manure with broadcast spreaders (P = 0.17), feeding of whole cottonseed or hulls (P = 0.17) or alfalfa (P = 0.15), and accessibility of feed to birds (P = 0.17) were identified as possible risk factors for C. jejuni infection. C. coli was detected in at least one animal in 19.4% of operations and 1.8% of individual cows (n = 2,085). At the herd level, use of broadcaster spreaders was not a risk factor for C. coli infection. For Arcobacter, cows from 71% of dairy operations (n = 31) and 14.3% of individual dairy cattle fecal samples (n = 1,682) were positive. At the herd level, for Arcobacter spp., feeding of alfalfa (P = 0.11) and use of individual waterers (P = 0.19) were protective. This is the first description of Arcobacter spp. in clinically healthy dairy cattle and the first attempt to correlate their presence with C. jejuni.
epidemiologic assessment for the transmission (8000 g, 5 min) and resuspended in 15 ml of PBS, with 5 of the organism to broiler chickens is needed to...spectrophotometrically (O.D.260 ). The DNA (2 uig) was digested (3-4 h, 37°C), Serotyping with BgIlI or HhaI in a 20 d reaction mixture in buffers Both the...original (0) and the recovered (R) Campylobacter uppliee by the manufacturer. After digestion , 5 pul of spp. strains were coded and sent to cooperating
Bates, C; Hiett, K L; Stern, N J
Campylobacter, a foodborne pathogen closely associated with poultry, is considered to be an important agent of human gastroenteritis in New Zealand. The pathways involved in the contamination of poultry flocks remain unclear; however, many vectors, such as insects, rodents, and wild birds, have been implicated. Infestation of poultry houses by insects, particularly darkling beetles (Alphitobius diaperinus), is difficult to control. Furthermore, darkling beetles are known vectors for a variety of pathogens that include Salmonella, infectious bursal disease virus, Aspergillus, Escherichia coli, and Marek's disease virus. In this investigation, the relationship between darkling beetles and Campylobacter contamination of poultry flocks was investigated. A New Zealand breeder flock and four of its progeny broiler flocks were included in the study. Samples of beetles and of intestinal excreta of the birds were cultured for the presence of Campylobacter spp. A subset of the recovered isolates was subsequently genotyped using flaA short variable region (SVR) DNA sequence analysis. A large number of Campylobacter subtypes were isolated, indicating that Campylobacter colonization of poultry is likely to arise from a number of different reservoirs. However, a set of genetically distinct isolates were found to be common to the broiler flocks and to the beetles. This research provides data that indicates that Alphitobius diaperinus may serve as a source of Campylobacter contamination of poultry. A more thorough understanding of the relationship between beetle infestation and the Campylobacter status of poultry flocks should enable progress in further development of biosecurity control measures.
Jonker, A; Picard, J A
Campylobacter jejuni is one of the leading causes of sporadic food-borne bacterial disease in humans. In intensive poultry and pig rearing systems the use of oral antibiotics is essential to maintain health. Consequently, there is a high risk for the thermophilic Campylobacter jejuni and C. coli resident in the intestinal tract of food animals to develop resistance to commonly used antibiotics. Contamination of meat or eggs with pathogenic strains of resistant Campylobacter could, therefore, result in a form of campylobacteriosis in humans that is difficult to treat. The aim of this investigation was to determine the antimicrobial susceptibility of thermophilic Campylobacter spp. isolated from pigs and poultry by the broth microdilution minimum inhibitory concentration (MIC) test. A total of 482 samples from the Western Cape and Gauteng provinces was collected and analysed. Thirty-eight Campylobacter isolates were obtained. Analysis of data revealed that C. jejuni strains mainly of poultry origin were more resistant to the fluoroquinolones, macrolides and tetracyclines and the C. coli strains were more resistant to the macrolides and lincosamides. Multi-resistance was also detected in 4 Campylobacter strains from the Western Cape. With the exception of tetracyclines, strains from high health Gauteng broiler farms were susceptible to antibiotics used to treat Campylobacter infections.
Zhao, S; Young, S R; Tong, E; Abbott, J W; Womack, N; Friedman, S L; McDermott, P F
The emergence of antimicrobial resistance in Campylobacter spp. has been a growing public health concern globally. The objectives of this study were to determine the prevalence, antimicrobial susceptibility, and genetic relatedness of Campylobacter spp. recovered by the National Antimicrobial Resistance Monitoring System (NARMS) retail meat program. Retail meat samples (n = 24,566) from 10 U.S. states collected between 2002 and 2007, consisting of 6,138 chicken breast, 6,109 ground turkey, 6,171 ground beef, and 6,148 pork chop samples, were analyzed. A total of 2,258 Campylobacter jejuni, 925 Campylobacter coli, and 7 Campylobacter lari isolates were identified. Chicken breast samples showed the highest contamination rate (49.9%), followed by ground turkey (1.6%), whereas both pork chops and ground beef had <0.5% contamination. The most common resistance was to doxycycline/tetracycline (46.6%), followed by nalidixic acid (18.5%), ciprofloxacin (17.4%), azithromycin and erythromycin (2.8%), telithromycin (2.4%), clindamycin (2.2%), and gentamicin (<0.1%). In a subset of isolates tested, no resistance to meropenem and florfenicol was seen. C. coli isolates showed higher resistance rates to antimicrobials, with the exception of doxycycline/tetracycline, than those seen for C. jejuni. Pulsed-field gel electrophoresis (PFGE) fingerprinting resulted in 1,226 PFGE profiles among the 2,318 isolates, with many clones being widely dispersed throughout the 6-year sampling period.
Maal-Bared, Rasha; Bartlett, Karen H; Bowie, William R; Hall, Eric R
Despite its relevance to public health, presence and concentrations of Campylobacter spp. in biofilms in natural aquatic environments has not been investigated. This study examined the occurrence of Campylobacter spp. in biofilms on a variety of surfaces (river rock, slate rock, wood, Lexan™, sandpaper, and sediment) and in water from December 2005 to December 2006 to find a substratum that facilitated campylobacters detection in natural aquatic environments. Samples were collected at four sites in an agricultural watershed (Elk Creek, British Columbia). Campylobacter spp. presence was determined using culturing methods. Correlations between chemical, physical and microbiological water quality parameters and Campylobacter spp. distribution on different surface types were also investigated. Campylobacter spp. had a prevalence of 13% in the wet season, but was not recovered in the dry season. Its prevalence was highest in sediment (27%), followed by slate rock (22%), Lexan and wood (13%), river rock (9%) and water (8%), respectively. No Campylobacter spp. was found in sandpaper biofilms. Several other criteria were used to assess substrata effectiveness, such as correlation amongst Campylobacter spp., indicator bacteria and water quality parameters, cost and availability of substratum, potential for standardizing substratum, ease of biofilm removal and probability of substratum loss in situ. Results show that sediment, slate rock or wood could be used as substrata for Campylobacter spp. monitoring. The study also highlights the potential use of nitrates and enterococci as faecal contamination indicators to protect public health.
Kim, Jinyong; Oh, Euna; Banting, Graham S.; Braithwaite, Shannon; Chui, Linda; Ashbolt, Nicholas J.; Neumann, Norman F.; Jeon, Byeonghwa
Campylobacter jejuni is one of the leading foodborne pathogens worldwide. C. jejuni is isolated from a wide range of foods, domestic animals, wildlife, and environmental sources. The currently available culture-based isolation methods are not highly effective for wastewater samples due to the low number of C. jejuni in the midst of competing bacteria. To detect and isolate C. jejuni from wastewater samples, in this study, we evaluated a few different enrichment conditions using five different antibiotics (i.e., cefoperazone, vancomycin, trimethoprim, polymyxin B, and rifampicin), to which C. jejuni is intrinsically resistant. The selectivity of each enrichment condition was measured with Ct value using quantitative real-time PCR, and multiplex PCR to determine Campylobacter species. In addition, the efficacy of Campylobacter isolation on different culture media after selective enrichment was examined by growing on Bolton and Preston agar plates. The addition of polymyxin B, rifampicin, or both to the Bolton selective supplements enhanced the selective isolation of C. jejuni. The results of 16S rDNA sequencing also revealed that Enterococcus spp. and Pseudomonas aeruginosa are major competing bacteria in the enrichment conditions. Although it is known to be difficult to isolate Campylobacter from samples with heavy contamination, this study well exhibited that the manipulation of antibiotic selective pressure improves the isolation efficiency of fastidious Campylobacter from wastewater. PMID:27617011
We analyzed a selection of 101 Campylobacter spp. isolates sampled from Grenada, Puerto Rico and Alabama in order to evaluate the discriminatory strength of four prominent molecular fingerprinting methods: restriction fragment length polymorphism of the flaA gene (flaA-RFLP), pulsed-field gel electr...
Yang, Rongchang; Jacobson, Caroline; Gardner, Graham; Carmichael, Ian; Campbell, Angus J D; Ryan, Una
Faecal excretion of Campylobacter spp. and Salmonella enterica in sheep in Australia was determined using a quantitative multiplex PCR (qPCR) targeting the Campylobacter spp. purine biosynthesis gene (PurA) and the S. enterica outer membrane protein (ompF). The mutiplex qPCR was specific and Campylobacter spp. and S. enterica were each detected with a sensitivity of 5 organisms/µL faecal DNA extract. This multiplex qPCR was used to determine the prevalence and concentration of Campylobacter spp. and S. enterica in 3412 faecal samples collected from 1189 lambs on eight farms across South Australia (n = 2 farms), New South Wales (n = 1), Victoria (n = 2) and Western Australia (n = 3) at three sampling periods (weaning, post-weaning and pre-slaughter). The overall prevalences of Campylobacter spp. and S. enterica were 13.3% and 5.0%, respectively, with the highest prevalence for Campylobacter spp. in South Australia and the highest prevalence for S. enterica in New South Wales. Campylobacter jejuni was the only Campylobacter sp. identified from a subset of 120 positive samples sequenced at the 16S locus. S. enterica serovar Typhimurium was the only serovar of S. enterica identified from a subset of 120 positive samples sequenced at the ompF locus. Across all states, Campylobacter spp. had the highest median bacterial concentration in faeces at weaning and post-weaning (medians of 3.4 × 10(6) and 1.1 × 10(5), respectively), whereas S. enterica had the highest median bacterial concentration at pre-slaughter (1.8 × 10(5)/g faeces).
Soonthornchaikul, Nantika; Garelick, Hemda
Campylobacter species have been recognized as the most commonly reported cause of bacterial gastroenteritis in humans. The increase of resistance rates to drugs of choice used for treatment in campylobacteriosis is becoming a public health concern. In parallel, the increased use of antimicrobials in aquaculture may lead to the emergence of resistant microorganisms and is likely to cause additional health risk to humans through food consumption. The study assesses the presence of antimicrobial resistance in Campylobacter species isolated from three groups of bivalve molluscs (bloody cockles, green mussels, and oysters) purchased from markets in Bangkok. Thirty samples were collected from each group. Susceptibility to three antimicrobials was determined using the Epsilometer test. Rates of erythromycin, nalidixic acid, and ciprofloxacin resistance in Campylobacter isolates were 72-84%, 28-40%, and 21-25%, respectively. There was no statistically significant difference in the prevalence of each antimicrobial resistance between the three groups. This study demonstrates a significant level of antimicrobial resistance in the Campylobacter spp. isolated from molluscs with a particular high rate of resistance to erythromycin. Consumption of raw molluscs contaminated with antimicrobial-resistant Campylobacter spp. may therefore result in resistant infections in humans.
Kashoma, Isaac P.; Kassem, Issmat I.; Kumar, Anand; Kessy, Beda M.; Gebreyes, Wondwossen; Kazwala, Rudovick R.; Rajashekara, Gireesh
Foodborne Campylobacter infections pose a serious threat to public health worldwide. However, the occurrence and characteristics of Campylobacter in food animals and products remain largely unknown in Tanzania. The objective of this study was to determine the prevalence, antibiotic resistance, and genetic profiles (sequence types, STs) of Campylobacter isolated from feces of pigs and dairy and beef cattle in Tanzania. Overall, 259 (~30%) of 864 samples were positive for Campylobacter spp, which were detected in 32.5, 35.4, and 19.6% of the pig, dairy, and beef cattle samples, respectively. Multiplex PCR analysis identified 64.5 and 29.3% of the Campylobacter isolates as C. coli and C. jejuni, respectively. The majority (91.9%) of the isolates from pig samples were identified as C. coli, while C. jejuni accounted for 65.5% of the isolates from cattle. Antimicrobial susceptibility testing using the disk diffusion assay and the broth microdilution method revealed resistance to: ampicillin (Amp) (70.3% and 75.7%, respectively), gentamicin (Gen) (1.8% and 12.6%), streptomycin (Str) (65.8 and 74.8%), erythromycin (Ery) (41.4 and 48.7%), tetracycline (Tet) (18.9 and 23.4%), and ciprofloxacin (Cip) (14.4 and 7.2%). Resistance to nalidixic acid (Nal) (39.6%), azithromycin (Azm) (13.5%), and chloramphenicol (Chl) (4.5%) was determined using the disk diffusion assay only, while resistance to tylosin (Tyl) (38.7%) was quantified using the broth microdilution method. Multilocus sequence typing of 111 Campylobacter isolates resulted in the identification of 48 STs (26 C. jejuni and 22 C. coli) of which seven were novel (six C. jejuni and one C. coli). Taken together, this study revealed the high prevalence, genetic diversity and antimicrobial resistance of Campylobacter in important food animals in Tanzania, which highlights the urgent need for the surveillance and control of Campylobacter in this country. PMID:26617582
The aim of the work was to collect, evaluate, summarize and compare heat resistance data reported for Campylobacter, Enterococcus, Escherichia, Listeria, Salmonella and Yersinia spp. The work was limited to resistance in liquids with pH values 6–8. Results obtained under similar experimental conditions were sought. Thermal destruction lines for the various bacterial groups studied were constructed using log10 D values and treatment temperatures. There was a good linear relationship between log10 D and temperature with Escherichia coli, listerias and salmonellas. For campylobacters, enterococci and yersinias the relationships were weaker but, nevertheless, present. Using the slopes of the lines and their 95% confidence limits, z values and their 95% confidence limits were calculated. z values were compared with z values obtained from reports. The equations for the lines were also used for calculation of predicted means of D values at various treatment temperatures. 95% confidence limits on predicted means of D values and on predicted individual D values were also calculated. Lines and values are shown in figures and tables. Differences in heat resistance noted between and within the bacterial groups studied are discussed. PMID:14650540
St-Pierre, Karen; Lévesque, Simon; Frost, Eric; Carrier, Nathalie; Arbeit, Robert D; Michaud, Sophie
This study aimed to assess the importance of quantitatively detecting Campylobacter spp. in environmental surface water. The prevalence and the quantity of Campylobacter spp., thermotolerant coliforms, and Escherichia coli in 2,471 samples collected weekly, over a 2-year period, from 13 rivers and 12 streams in the Eastern Townships, Québec, Canada, were determined. Overall, 1,071 (43%), 1,481 (60%), and 1,463 (59%) samples were positive for Campylobacter spp., thermotolerant coliforms, and E. coli, respectively. There were weak correlations between the weekly distributions of Campylobacter spp. and thermotolerant coliforms (Spearman's rho coefficient = 0.27; P = 0.008) and between the quantitative levels of the two classes of organisms (Kendall tau-b correlation coefficient = 0.233; P < 0.0001). Well water samples from the Eastern Townships were also tested. Five (10%) of 53 samples from private surface wells were positive for Campylobacter jejuni, of which only 2 were positive for thermotolerant coliforms. These findings suggest that microbial monitoring of raw water by using only fecal indicator organisms is not sufficient for assessing the occurrence or the load of thermophilic Campylobacter spp. Insights into the role of environmental water as sources for sporadic Campylobacter infection will require genus-specific monitoring techniques.
Caner, Vildan; Cokal, Yavuz; Cetin, Cengiz; Sen, Aysin; Karagenc, Nedim
A total of 190 Campylobacter spp. isolates, of which 34 gave the result of very weak activity, and 156 gave the negative activity in the test for hippurate hydrolysis were characterized. The genomic DNA was isolated from a fresh culture of each isolate and the real-time PCR, targeting the hipO gene, was used to confirm the species distribution of Campylobacter isolates. The hipO gene was detected in 17 isolates (11%) within the total of 156 negative isolates for hippurate hydrolysis. Out of 34 isolates with very weak activity, 19 isolates (56%) were also found to be positive for hipO gene and characterized as C. jejuni. The real-time PCR assay used in this study could be employed for more accurate diagnosis of Campylobacter infections at species level after the biochemical characterization based on hippuricase activity of the isolates. This could also provide important data for the epidemiology of infections associated with these zoonotic pathogens.
Jonsson, M E; Norström, M; Sandberg, M; Ersbøll, A K; Hofshagen, M
This study was performed to investigate space-time patterns of Campylobacter spp. colonization in broiler flocks in Norway. Data on the Campylobacter spp. status at the time of slaughter of 16 054 broiler flocks from 580 farms between 2002 and 2006 was included in the study. Spatial relative risk maps together with maps of space-time clustering were generated, the latter by using spatial scan statistics. These maps identified the same areas almost every year where there was a higher risk for a broiler flock to test positive for Campylobacter spp. during the summer months. A modified K-function analysis showed significant clustering at distances between 2.5 and 4 km within different years. The identification of geographical areas with higher risk for Campylobacter spp. colonization in broilers indicates that there are risk factors associated with Campylobacter spp. colonization in broiler flocks varying with region and time, e.g. climate, landscape or geography. These need to be further explored. The results also showed clustering at shorter distances indicating that there are risk factors for Campylobacter spp. acting in a more narrow scale as well.
Kashoma, Isaac P; Kassem, Issmat I; John, Julius; Kessy, Beda M; Gebreyes, Wondwossen; Kazwala, Rudovick R; Rajashekara, Gireesh
Campylobacter species are commonly transmitted to humans through consumption of contaminated foods such as milk and meat. The aim of this study was to investigate the prevalence, antimicrobial resistance, and genetic determinants of resistance of Campylobacter isolated from raw milk and beef carcasses in Tanzania. The antimicrobial resistance genes tested included blaOXA-61 (ampicillin), aph-3-1 (aminoglycoside), tet(O) (tetracycline), and cmeB (multi-drug efflux pump). The prevalence of Campylobacter was 9.5% in beef carcasses and 13.4% in raw milk, respectively. Using multiplex-polymerase chain reaction (PCR), we identified 58.1% of the isolates as Campylobacter jejuni, 30.7% as Campylobacter coli, and 9.7% as other Campylobacter spp. One isolate (1.6%) was positive for both C. jejuni and C. coli specific PCR. Antimicrobial susceptibility testing using the disk diffusion assay and the broth microdilution method showed resistance to: ampicillin (63% and 94.1%), ciprofloxacin (9.3% and 11.8%), erythromycin (53.7% and 70.6%), gentamicin (0% and 15.7%), streptomycin (35.2% and 84.3%), and tetracycline (18.5% and 17.7%), respectively. Resistance to azithromycin (42.6%), nalidixic acid (64.8%), and chloramphenicol (13%) was determined using the disk diffusion assay only, while resistance to tylosin (90.2%) was quantified using the broth microdilution method. The blaOXA-61 (52.6% and 28.1%), cmeB (26.3% and 31.3%), tet(O) (26.3% and 31.3%), and aph-3-1 (5.3% and 3.0%) were detected in C. coli and C. jejuni. These findings highlight the extent of antimicrobial resistance in Campylobacter occurring in important foods in Tanzania. The potential risks to consumers emphasize the need for adequate control approaches, including the prudent use of antimicrobials to minimize the spread of antimicrobial-resistant Campylobacter.
Ganan, M; Carrascosa, A V; Martínez-Rodríguez, A J
The antimicrobial activities of three chitosans with different molecular masses against six gram-negative and three gram-positive bacteria were examined. Campylobacter spp. were the microorganisms most sensitive to chitosan, regardless of their molecular mass. The MIC of chitosan for Campylobacter ranged from 0.005 to 0.05%, demonstrating the global sensitivity of campylobacters to chitosan. Chitosan caused a loss in the membrane integrity of Campylobacter, measured as an increase in cell fluorescence due to the uptake of propidium iodide, a dye that is normally excluded from cells with intact membranes. As cells entered the stationary phase, there was a change in cell membrane resistance toward a loss of integrity caused by chitosan. This study demonstrates that chitosans could be a promising antimicrobial to control Campylobacter.
Cardinale, E; Tall, F; Guèye, E F; Cisse, M; Salvat, G
Our objective was to identify the risk factors for Campylobacter infection in Senegalese broiler flocks. Seventy broiler farms were studied around Dakar from January 2000 to December 2001 around Dakar. A questionnaire was administered to the farmers, and samples of fresh droppings were taken to assess the flocks' Campylobacter status. About 63% of the flocks were infected by Campylobacter spp.; Campylobacter jejuni was the most-prevalent species (P < 0.05). An elevated risk of Campylobacter infection was associated with other animals (mainly laying hens, cattle and sheep) being bred in the farm, the farm staff not wearing their work clothing exclusively in the poultry houses, uncemented poultry-house floors and the use of cartons that transport chicks from the hatchery to the farm as feed plates (rather than specifically designed feed plates). Alternatively, thorough cleaning and disinfection of poultry-house surroundings and manure disposal outside the farm were associated with decreased flock risk.
Gebhart, C J; Ward, G E; Kurtz, H J
The in vitro activities of 47 antimicrobial agents against 30 isolates of Campylobacter species from pigs were determined by the agar dilution technique. The isolates were obtained from pigs with proliferative enteritis and included 10 strains each of Campylobacter coli, Campylobacter sputorum subsp. mucosalis, and "Campylobacter hyointestinalis Gebhart et al." (this name is not on the Approved Lists). Carbadox, furazolidone, nitrofurantoin, gentamicin, and dimetridazole were the most active drugs, inhibiting all three Campylobacter species with a MIC for 50% of the isolates of 2 micrograms/ml or less. Trimethoprim-sulfamethoxazole, cefazolin, sulfachloropyridazine, novobiocin, vancomycin, sulfathiazole, cyclohexamide, bacitracin, p-arsanilic acid, and colistin were the least active, with MICs for 50% of the isolates ranging from 16 to greater than or equal to 128 micrograms/ml. PMID:3985597
Caballero, Moisés; Rivera, Isabel; Jara, Luis M; Ulloa-Stanojlovic, Francisco M; Shiva, Carlos
Feral pigeons (Columbia livia) live in close contact with humans and other animals. They can transmit potentially pathogenic and zoonotic agents. The objective of this study was to isolate and detect strains of diarrheagenic Escherichia coli and Campylobacter jejuni of urban feral pigeons from an area of Lima, Peru. Fresh dropping samples from urban parks were collected for microbiological isolation of E. coli strains in selective agar, and Campylobacter by filtration method. Molecular identification of diarrheagenic pathotypes of E.coli and Campylobacter jejuni was performed by PCR. Twenty-two parks were sampled and 16 colonies of Campylobacter spp. were isolated. The 100% of isolates were identified as Campylobacter jejuni. Furthermore, 102 colonies of E. coli were isolated and the 5.88% resulted as Enteropathogenic (EPEC) type and 0.98% as Shiga toxin-producing E. coli (STEC). The urban feral pigeons of Lima in Peru can act as a reservoir or carriers of zoonotic potentially pathogenic enteric agents.
Templeton, Jillian M; De Jong, Amanda J; Blackall, P J; Miflin, Jeanette K
Campylobacter infection is the most frequently reported notifiable food-borne disease in humans in Australia. Our studies investigated the persistence of Campylobacter spp. in or on darkling beetles (Alphitobius diaperinus) and their larvae. Our results in analyses with chickens confirm that, unless very short turnaround times are used (<72 h), beetles colonized in one production cycle (i.e., one batch of chickens) are most unlikely to still be colonized during the next cycle of chickens.
Templeton, Jillian M.; De Jong, Amanda J.; Blackall, P. J.; Miflin, Jeanette K.
Campylobacter infection is the most frequently reported notifiable food-borne disease in humans in Australia. Our studies investigated the persistence of Campylobacter spp. in or on darkling beetles (Alphitobius diaperinus) and their larvae. Our results in analyses with chickens confirm that, unless very short turnaround times are used (<72 h), beetles colonized in one production cycle (i.e., one batch of chickens) are most unlikely to still be colonized during the next cycle of chickens. PMID:17012593
Henry, Isabelle; Reichardt, Jef; Denis, Martine; Cardinale, Eric
Our objectives were to determine Campylobacter prevalence in broiler chicken flocks in Reunion Island and to define specific practices associated with the presence of Campylobacter spp. Infection in Reunionese broiler flocks. Fifty broiler flocks were studied in Reunion Island from May 2007 to February 2009. A questionnaire was submitted to the farmers and samples of fresh droppings were collected to assess the flock's Campylobacter status. Fifty four percent of the flocks were infected by Campylobacter spp.: 30% (95% CI: 28.71-31.29) were infected with Campylobacter coli and 17% (95% CI: 15.95-18.05) with Campylobacter jejuni; only 7% (95% CI: 6.28-7.72) were infected by both species at the same time. Several poultry houses in the farm (OR=11.2; [1.05-92]) and cleaning without any detergent (OR=13.1; [2.1-78.3]) increased the risk of Campylobacter infection. A distance higher than 500 m between broiler farms (OR=0.27; [0.1-0.8]) and use of disinfectant during the rearing period decreased this risk of infection (OR=0.15; [0.1-0.75]).
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.
Hill, Janet E; Paccagnella, Ana; Law, Kee; Melito, Pasquale L; Woodward, David L; Price, Lawrence; Leung, Amy H; Ng, Lai-King; Hemmingsen, Sean M; Goh, Swee Han
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.
Campylobacter spp. is a Gram negative bacterium and is the major cause of foodborne gastroenteritis worldwide. The microaerophilic nature of Campylobacter and its requirement of ~5% O2 for growth have complicated its recovery from foods. This is achieved with the addition to the enrichment media of ...
Dickins, M Avery; Franklin, Sharon; Stefanova, Rossina; Schutze, Gordon E; Eisenach, Kathleen D; Wesley, Irene; Cave, M Donald
Campylobacter spp. are a major contaminant of poultry. Eating undercooked chicken and handling raw poultry have been identified as risk factors for campylobacteriosis in humans. Previous studies have found Campylobacter spp. on 90% of poultry carcasses. In the present study, pulsed-field gel electrophoresis (PFGE) was used to assess the genetic diversity of strains on retail poultry carcasses. PFGE patterns of isolates from campylobacteriosis cases were compared to those from the poultry isolates. Over a 1-year study period (March 2000 through February 2001), whole fresh young chickens (n = 72) were obtained from three retail outlets in an urban community in the south-central United States. Campylobacter spp. were isolated from 82% of these carcasses. Strains (n = 70) were defined on the basis of their PFGE pattern. Sixty-seven percent of the carcasses from which Campylobacter spp. were isolated were contaminated with more than one PFGE-distinguishable strain. During the 1-year study period, most of the PFGE patterns (59%) were limited to isolates obtained from a single carcass. Forty-one percent of the PFGE-distinguishable strains were recovered from more than one carcass. Ninety-seven percent of the carcasses contaminated with the same strain were purchased at the same time from the same store. To examine the degree of genetic stability, four strains were followed in vitro over an estimated 1,000 doublings. The PFGE pattern of one of these isolates underwent minor changes during in vitro growth. The data indicate extensive variability in the PFGE patterns of Campylobacter spp. isolated from humans and from poultry carcasses. In spite of difficulties caused by such diversity and the fact that some carcasses are contaminated with more than one strain, the pattern variation provides a useful method for linking a particular strain to its source.
Campylobacter spp., considered a leading bacterial etiology of acute gastroenteritis in humans, is commonly associated with poultry. However, the factors involved in colonization of poultry with Campylobacter spp. remain unclear. Determination of colonization-associated factors should facilitate ou...
Nielsen, Hans Linde; Ejlertsen, Tove; Nielsen, Henrik
A total of 5963 diarrheic stool samples were cultivated for Campylobacter spp. with use of modified charcoal cefoperazone deoxycholate agar (mCCDA) plates as well as a polycarbonate (PC) filter technique on blood agar plates. A total of 376 Campylobacter jejuni/coli were isolated from both PC and mCCDA. Six and three were isolated from PC and mCCDA only, respectively (P = ns). The PC technique is noninferior to mCCDA for isolation of C. jejuni/coli.
Feodoroff, Benjamin; de Haan, Caroline P A; Ellström, Patrik; Sarna, Seppo; Hänninen, Marja-Liisa; Rautelin, Hilpi
Campylobacter jejuni bacteria are highly diverse enteropathogens. Seventy-three C. jejuni isolates from blood collected in Finland were analyzed by multilocus sequence typing and serum resistance. Approximately half of the isolates belonged to the otherwise uncommon sequence type 677 clonal complex. Isolates of this clonal complex were more resistant than other isolates to human serum.
The unacceptably high frequency of Campylobacter jejuni transmission from poultry to humans encourages scientists to consider and create alternative intervention strategies to control the pathogen in poultry production. Extremely high numbers of Campylobacter (often >108 cfu/g of poultry intestinal...
The unacceptably high frequency of Campylobacter jejuni transmission from poultry to humans encourages scientists to consider and create alternative intervention strategies to control the pathogen in poultry production. Extremely high numbers of Campylobacter (often >108 cfu/g of poultry intestinal...
Campylobacter continues to be one of the most common bacterial causes of diarrheal illness in the United States and worldwide. Infection with Campylobacter causes a spectrum of diseases including acute enteritis, extraintestinal infections, and postinfectious complications. The most common species of Campylobacter associated with human illness is Campylobacter jejuni, but other Campylobacter species can also cause human infections. This comprehensive review includes discussion of the taxonomy, clinical manifestations of infection, epidemiology and the different methods of laboratory detection of Campylobacter.
To determine if Campylobacter isolation method influenced antimicrobial susceptibility results, the minimum inhibitory concentrations (MIC) of nine antimicrobials were compared for 291 pairs of Campylobacter isolates recovered from chicken carcass rinse samples using direct plating and an enrichment...
Christidis, T; Pintar, K D M; Butler, A J; Nesbitt, A; Thomas, M K; Marshall, B; Pollari, F
Campylobacteriosis is the leading bacterial gastrointestinal disease internationally, contributing significantly to the enteric illness burden. Cases have been associated with the consumption of raw milk, a behavior that has garnered attention recently. Estimates of the prevalence and levels of Campylobacter spp. in raw milk are lacking, which hinders risk assessment attempts. This article is a systematic review and meta-analysis of reported prevalence and levels of zoonotic Campylobacter spp. in the raw milk of cows, goats, and sheep in Canada, the United States, Europe, Australia, and New Zealand. The relevant literature was reviewed, and trained reviewers examined the results for inclusion of articles in the meta-analysis. Relevant data (prevalence and/or level of Campylobacter in raw milk, country of origin, animal species, sample source, Campylobacter species identified, etc.) were extracted, and a meta-analysis was performed in Stata v. 12 (Metaprop command). The weighted mean prevalence of Campylobacter spp. in raw milk samples was 1.18%. Subgroup analyses were conducted to examine how prevalence varied by study characteristics, with the highest prevalence values in studies from the United Kingdom (by country, 6.4%), about cows (by animal species, 1.3%), and including samples taken from inline filters (by sample source, 1.75%) and in studies that included species that are not pathogenic to humans (by Campylobacter species, 1.14%). Two articles each included a single Campylobacter level, 0.16 ± 0.3 and approximately 0.047 most probable number per ml. Despite a relatively low prevalence, consumption of raw milk is inherently risky because no treatment has been used to inactivate pathogens. This potential risk further supports maintaining regulations to limit the sales of raw milk.
Turowski, E E; Shen, Z; Ducore, R M; Parry, N M A; Kirega, A; Dewhirst, F E; Fox, J G
Routine necropsies of 27 asymptomatic juvenile chinchillas revealed a high prevalence of gastric ulcers with microscopic lymphoplasmacytic gastroenteritis and typhlocolitis. Polymerase chain reaction (PCR) analysis using Campylobacter genus-specific partial 16S rRNA primers revealed the presence of Campylobacter spp. DNA in the faeces of 12 of 27 animals (44.4%). Species-specific partial 16S rRNA PCR and sequencing confirmed that these animals were colonized with Campylobacter lanienae, a gram-negative, microaerophilic bacterium that was first identified on routine faecal screening of slaughterhouse employees and subsequently isolated from faeces of livestock. Campylobacter lanienae was isolated from the faeces of six PCR-positive animals and identified with species-specific PCR and full 16S rRNA sequencing. Phylogenetic analysis showed that these isolates clustered with C. lanienae strain NCTC 13004. PCR analysis of DNA extracted from gastrointestinal tissues revealed the presence of C. lanienae DNA in the caecum and colon of these chinchillas. Gastrointestinal lesions were scored and compared between C. lanienae-positive and C. lanienae-negative animals. There was no correlation between colonization status and lesion severity in the stomach, liver, duodenum, or colon. Possible routes of C. lanienae infection in chinchillas could include waterborne transmission and faecal-oral transmission from wild mice and rats or livestock. Based on these findings, the authors conclude that C. lanienae colonizes the lower bowel of chinchillas in the absence of clinical disease. This is the first report of C. lanienae in any rodent species. Campylobacter lanienae isolates from different mammalian species demonstrate heterogeneity by 16S rRNA sequence comparison. Analysis using rpoB suggests that isolates and clones currently identified as C. lanienae may represent multiple species or subspecies.
Ryu, Hodon; Grond, Kirsten; Verheijen, Bram; Elk, Michael; Buehler, Deborah M.
Using 16S rRNA gene sequencing analysis, we examined the bacterial diversity and the presence of opportunistic bacterial pathogens (i.e., Campylobacter and Helicobacter) in red knot (Calidris canutus; n = 40), ruddy turnstone (Arenaria interpres; n = 35), and semipalmated sandpiper (Calidris pusilla; n = 22) fecal samples collected during a migratory stopover in Delaware Bay. Additionally, we studied the occurrence of Campylobacter spp., enterococci, and waterfowl fecal source markers using quantitative PCR (qPCR) assays. Of 3,889 16S rRNA clone sequences analyzed, the bacterial community was mostly composed of Bacilli (63.5%), Fusobacteria (12.7%), Epsilonproteobacteria (6.5%), and Clostridia (5.8%). When epsilonproteobacterium-specific 23S rRNA gene clone libraries (i.e., 1,414 sequences) were analyzed, the sequences were identified as Campylobacter (82.3%) or Helicobacter (17.7%) spp. Specifically, 38.4%, 10.1%, and 26.0% of clone sequences were identified as C. lari (>99% sequence identity) in ruddy turnstone, red knot, and semipalmated sandpiper clone libraries, respectively. Other pathogenic species of Campylobacter, such as C. jejuni and C. coli, were not detected in excreta of any of the three bird species. Most Helicobacter-like sequences identified were closely related to H. pametensis (>99% sequence identity) and H. anseris (92% sequence identity). qPCR results showed that the occurrence and abundance of Campylobacter spp. was relatively high compared to those of fecal indicator bacteria, such as Enterococcus spp., E. faecalis, and Catellicoccus marimammalium. Overall, the results provide insights into the complexity of the shorebird gut microbial community and suggest that these migratory birds are important reservoirs of pathogenic Campylobacter species. PMID:24413599
Campylobacter spp. is an important zoonotic microaerophilic bacterial pathogen that caused the majority of US outbreaks associated with nonpasteurized milk from 2007 to 2012. Bulk tank milk and milk filter samples were collected from 236 dairy operations in 17 top dairy states from March through Jul...
Morant-Miñana, M Carmen; Elizalde, J
Campylobacter spp. are responsible for acute bacterial diseases in human worldwide. Nowadays campilobacteriosis is considered the most common foodborne illness in the European Union. In this paper the first electrochemical genosensor based on thin-film gold electrodes deposited onto Cyclo Olefin Polymer (COP) substrates was fabricated for the detection of Campylobacter spp in food matrices. The sensing element is characterized by several surface techniques and the sensitivity of the biosensor have been studied. A good linear relationship was obtained for the concentrations of PCR amplicon of Campylobacter spp. between 1 and 25 nM with a limit of detection (LOD) of 90 pM. Real samples have been validated with poultry meat samples and results were comparable with the PCR product samples. This is the last step for the fabrication of a Lab on a Chip (LOC), a biodevice integrating DNA sensor technology into microfluidic system, believed to perform an automated and complete assay, including sample preparation, PCR amplification, and electrochemical detection of Campylobacter spp. in raw poultry meat samples.
Prevalence of Shiga toxin-producing Escherichia coli, Salmonella spp. and Campylobacter spp. in large game animals intended for consumption: relationship with management practices and livestock influence.
Díaz-Sánchez, S; Sánchez, S; Herrera-León, S; Porrero, C; Blanco, J; Dahbi, G; Blanco, J E; Mora, A; Mateo, R; Hanning, I; Vidal, D
Although wild ruminants have been identified as reservoirs of Shiga-toxin producing Escherichia coli (STEC), little information is available concerning the role of Salmonella spp. and Campylobacter spp. in large game species. We evaluated the presence of these pathogens in faeces (N=574) and carcasses (N=585) sampled from red deer (N=295), wild boar (N=333) and other ungulates (fallow deer, mouflon) (N=9). Animal sampling was done in situ from 33 hunting estates during two hunting seasons. Salmonella spp. and Campylobacter spp. strains associated with human campylobacteriosis were infrequently detected indicating that both pathogens had a limited zoonotic risk in our study area. The overall STEC prevalence in animals was 21% (134/637), being significantly higher in faeces from red deer (90 out of 264). A total of 58 isolates were serotyped. Serotypes O146:H- and O27:H30 were the most frequent in red deer and the majority of isolates from red deer and wild boar were from serotypes previously found in STEC strains associated with human infection, including the serotype O157:H7. The STEC prevalence in red deer faeces was significantly higher with the presence of livestock (p<0, 01) where high densities of red deer (p<0.001) were present. To the best of our knowledge, this is the first study reporting the occurrence of Salmonella spp. and STEC in carcasses of large game animals. Furthermore, this study confirmed by pulsed-field gel electrophoresis (PFGE) that cross contamination of STEC during carcass dressing occurred, implying the likelihood of these pathogens entering into the food chain.
Stone, Diana M; Chander, Yogesh; Bekele, Aschalew Z; Goyal, Sagar M; Hariharan, Harry; Tiwari, Keshaw; Chikweto, Alfred; Sharma, Ravindra
Rectal swabs from 155 sheep and 252 goats from Grenada were evaluated to determine the prevalence of Campylobacter spp., antibiotic resistance, and multilocus sequence types. Fifteen Campylobacter isolates were obtained (14 C. jejuni and 1 C. coli). The prevalence (3.7%) did not differ significantly between sheep (4.5%) and goats (3.2%). Among the seven antimicrobials tested, resistance was only detected for tetracycline (30.8%) and metronidazole (38.5%). Campylobacter isolates showed no significant difference between sheep and goats for type of antimicrobial resistance or percent of resistant isolates. Twelve of the isolates were successfully genotyped consisting of four recognized clonal complexes and three novel sequence types. Importantly, one isolate from one goat was identified as the C. jejuni sequence type-8, a zoonotic and tetracycline-resistant clone reported to be a highly virulent clone associated with ovine abortion in the USA. Although most samples were from comingled sheep and goat production units, there were no shared sequence types between these two host species. None of the sequence types identified in this study have previously been reported in poultry in Grenada, suggesting sheep- and goat-specific Campylobacter clones in Grenada. This is the first report of genotyping of Campylobacter isolates from sheep and goats in the Eastern Caribbean.
Fernando, Dinesh M.; Khan, Izhar U. H.; Patidar, Rakesh; Lapen, David R.; Talbot, Guylaine; Topp, Edward; Kumar, Ayush
Acinetobacter baumannii, a Gram-negative opportunistic pathogen, is known to cause multidrug resistant infections. This organism has primarily been isolated from clinical environments and its environmental reservoirs remain largely unknown. In the present study, we recovered seven isolates of A. baumannii growing under conditions selective for Campylobacter spp. (microaerophilic at 42°C and in the presence of antibiotics) from dairy cattle manure storage tank or surface water impacted by livestock effluents. Antibiotic susceptibility tests revealed that all of these isolates were less susceptible to at least two different clinically relevant antibiotics, compared to the type strain A. baumannii ATCC17978. Expression of resistance-nodulation-division efflux pumps, an important mechanism of intrinsic resistance in these organisms, was analyzed, and adeB was found to be overexpressed in one and adeJ was overexpressed in three isolates. Comparison of these isolates using genomic DNA Macro-Restriction Fragment Pattern Analysis (MRFPA) revealed relatively low relatedness among themselves or with some of the clinical isolates from previous studies. This study suggests that A. baumannii isolates are capable of growing under selective conditions for Campylobacter spp. and that this organism can be present in manure and water. PMID:27917170
Harrison, James W.; Dung, Tran Thi Ngoc; Siddiqui, Fariha; Korbrisate, Sunee; Bukhari, Habib; Tra, My Phan Vu; Hoang, Nguyen Van Minh; Carrique-Mas, Juan; Bryant, Juliet; Campbell, James I.; Studholme, David J.; Wren, Brendan W.; Baker, Stephen; Titball, Richard W.
A novel protein translocation system, the type-6 secretion system (T6SS), may play a role in virulence of Campylobacter jejuni. We investigated 181 C. jejuni isolates from humans, chickens, and environmental sources in Vietnam, Thailand, Pakistan, and the United Kingdom for T6SS. The marker was most prevalent in human and chicken isolates from Vietnam. PMID:24856088
Harrison, James W; Dung, Tran Thi Ngoc; Siddiqui, Fariha; Korbrisate, Sunee; Bukhari, Habib; Tra, My Phan Vu; Hoang, Nguyen Van Minh; Carrique-Mas, Juan; Bryant, Juliet; Campbell, James I; Studholme, David J; Wren, Brendan W; Baker, Stephen; Titball, Richard W; Champion, Olivia L
A novel protein translocation system, the type-6 secretion system (T6SS), may play a role in virulence of Campylobacter jejuni. We investigated 181 C. jejuni isolates from humans, chickens, and environmental sources in Vietnam, Thailand, Pakistan, and the United Kingdom for T6SS. The marker was most prevalent in human and chicken isolates from Vietnam.
Olkkola, Satu; Kovanen, Sara; Roine, Johanna; Hänninen, Marja-Liisa; Hielm-Björkman, Anna; Kivistö, Rauni
In recent years, increasing numbers of consumers have become interested in feeding raw food for their pet dogs as opposed to commercial dry food, in the belief of health advantages. However, raw meat and internal organs, possibly contaminated by pathogens such as Campylobacter spp., may pose a risk of transmission of zoonoses to the pet owners. Campylobacter jejuni is the leading cause of bacterial gastroenteritis in humans but C. upsaliensis has also been associated with human disease. In this study we investigated the effect of different feeding strategies on the prevalence of Campylobacter spp. in Finnish dogs. We further characterized the isolates using multilocus sequence typing (MLST), whole-genome (wg) MLST and antimicrobial susceptibility testing. Dogs were sampled before and after a feeding period consisting of commercial raw feed or dry pellet feed. Altogether 56% (20/36) of the dogs yielded at least one Campylobacter-positive fecal sample. C. upsaliensis was the major species detected from 39% of the dogs before and 30% after the feeding period. Two C. jejuni isolates were recovered, both from raw-fed dogs after the dietary regimen. The isolates represented the same genotype (ST-1326), suggesting a common infection source. However, no statistically significant correlation was found between the feeding strategies and Campylobacter spp. carriage. The global genealogy of MLST types of dog and human C. upsaliensis isolates revealed weakly clonal population structure as most STs were widely dispersed. Major antimicrobial resistance among C. upsaliensis isolates was against streptomycin (STR MIC > 4 mg/l). Apart from that, all isolates were highly susceptible against the antimicrobials tested. Mutations were found in the genes rpsL or rpsL and rsmG in streptomycin resistant isolates. In conclusion, increasing trend to feed dogs with raw meat warrants more studies to evaluate the risk associated with raw feeding of pets in transmission of zoonoses to humans.
Roine, Johanna; Hänninen, Marja-Liisa; Hielm-Björkman, Anna; Kivistö, Rauni
In recent years, increasing numbers of consumers have become interested in feeding raw food for their pet dogs as opposed to commercial dry food, in the belief of health advantages. However, raw meat and internal organs, possibly contaminated by pathogens such as Campylobacter spp., may pose a risk of transmission of zoonoses to the pet owners. Campylobacter jejuni is the leading cause of bacterial gastroenteritis in humans but C. upsaliensis has also been associated with human disease. In this study we investigated the effect of different feeding strategies on the prevalence of Campylobacter spp. in Finnish dogs. We further characterized the isolates using multilocus sequence typing (MLST), whole-genome (wg) MLST and antimicrobial susceptibility testing. Dogs were sampled before and after a feeding period consisting of commercial raw feed or dry pellet feed. Altogether 56% (20/36) of the dogs yielded at least one Campylobacter-positive fecal sample. C. upsaliensis was the major species detected from 39% of the dogs before and 30% after the feeding period. Two C. jejuni isolates were recovered, both from raw-fed dogs after the dietary regimen. The isolates represented the same genotype (ST-1326), suggesting a common infection source. However, no statistically significant correlation was found between the feeding strategies and Campylobacter spp. carriage. The global genealogy of MLST types of dog and human C. upsaliensis isolates revealed weakly clonal population structure as most STs were widely dispersed. Major antimicrobial resistance among C. upsaliensis isolates was against streptomycin (STR MIC > 4mg/l). Apart from that, all isolates were highly susceptible against the antimicrobials tested. Mutations were found in the genes rpsL or rpsL and rsmG in streptomycin resistant isolates. In conclusion, increasing trend to feed dogs with raw meat warrants more studies to evaluate the risk associated with raw feeding of pets in transmission of zoonoses to humans
Campylobacter spp. are the most commonly reported bacterial cause of acute diarrheal disease in humans throughout the world. Traditional cultural methods for the detection and quantitation of Campylobacterspp. are slow and tedious; therefore, specific, sensitive, and rapid methods for campylobacters are needed to collect sufficient data for risk assessment and food safety policy development. We developed several rapid methods based on polymerase chain reaction (PCR), DNA hybridization, hydrophobic grid membrane filters (HGMFs), and enzyme immunoassays (EIAs). A PCR assay targeting C. jejuni, combined with a simple sample preparation procedure, detects as few as 0.3 most probable number (MPN)/mL C. jejuni in naturally contaminated chicken rinses after 20-24 h enrichment. An HGMF-EIA method using a commercial polyclonal antibody for Campylobacter detects and enumerates thermophilic Campylobacter spp. from spiked chicken rinse and milk, and naturally contaminated chicken rinses. A C. jejuni-specific probe in an HGMF-DNA hybridization protocol specifically detects and quantitates C. jejuni in food samples. A dot-blot EIA combined with an MPN procedure quantitates thermophilic campylobacters from samples that might be difficult to filter through HGMFs.
Klein, Günter; Jansen, Wiebke; Kittler, Sophie; Reich, Felix
In contrast to other foodborne zoonotic agents an elimination of Campylobacter spp. from animal production, especially poultry production, seems not to be feasible. Therefore mitigation strategies focus on reduction of the Campylobacter spp. concentration in primary production and further minimalisation during processing. In primary production biosecurity measures (incl. hygiene barriers and restricted access) are the methods applied most commonly and most effectively so far. Experimental approaches and few field trials also showed that bacteriophages, electrolyzed oxidizing water, organic acids or medium chain fatty acids (applied via drinking water) are also effective in reducing Campylobacter prevalence and/or concentration However this reduction cannot be transferred in all cases to the situation in the slaughterhouse. Therefore additional measures have to be taken in account in the slaughterhouse to prevent cross-contamination. Logistic or scheduled slaughter can prevent cross-contamination but cannot further reduce Campylobacter concentration. Process parameters like elevated scalding temperature can contribute to such a reduction, but may also alter the product quality. Therefore no single pre- or harvest measure is sufficient for the reduction of Campylobacter concentration, but a combination of measures in both production levels is needed.
Kassa, Tesfaye; Gebre-Selassie, Solomon; Asrat, Daniel
Thermotolerant Campylobacter spp. are frequent causes of diarrhoea in humans worldwide mostly originating from poultry. It has been suggested that extensive veterinary use of antibiotics is largely responsible for resistance in human isolates. During a 4-month period from January to April 2004, 192 Campylobacter spp. were isolated from fecal samples of 485 healthy food animals. The in vitro susceptibility to 12 antibiotics was determined by the agar disk diffusion method. Among the 192 Campylobacter spp. isolated, 135 (70.3%) were identified to be C. jejuni, 51 (26.6%) were C. coli and 6 (3.1%) were C. lari. C. jejuni was the most prevalent species in chickens (80.8%) versus 16.2% C. coli and 3.0% C. lari. All isolates found in pigs were C. coli. All strains were sensitive to chloramphenicol and ciprofloxacin and all were resistant to cephalothin. More than 90% of the strains were sensitive to clindamycin, erythromycin, gentamicin, nalidixic acid, norfloxacin, streptomycin and tetracycline. Resistance was found against ampicillin in 20% and trimethoprim-sulphamethoxazole in 37.5%. Resistance was not statistically different among C. jejuni, C. coli and C. lari (p>0.05). Multidrug resistance to two or more drugs was detected in 14.5% of strains. In conclusion, the study showed that antimicrobial resistance is found only at relatively low frequencies for most antimicrobial agents tested except for ampicillin and trimethoprim-sulphamethoxazole. The low percentages of resistance to most antimicrobial agents tested in this study may be the result of low/no usage of these agents as a growth promoters or treatment in the Ethiopian animal farm setting. The detection of multidrug resistant isolates may pose a threat to humans and further limits therapeutic options.
Kashoma, Isaac P.; Kassem, Issmat I.; John, Julius; Kessy, Beda M.; Gebreyes, Wondwossen; Kazwala, Rudovick R.
Campylobacter species are commonly transmitted to humans through consumption of contaminated foods such as milk and meat. The aim of this study was to investigate the prevalence, antimicrobial resistance, and genetic determinants of resistance of Campylobacter isolated from raw milk and beef carcasses in Tanzania. The antimicrobial resistance genes tested included blaOXA-61 (ampicillin), aph-3-1 (aminoglycoside), tet(O) (tetracycline), and cmeB (multi-drug efflux pump). The prevalence of Campylobacter was 9.5% in beef carcasses and 13.4% in raw milk, respectively. Using multiplex-polymerase chain reaction (PCR), we identified 58.1% of the isolates as Campylobacter jejuni, 30.7% as Campylobacter coli, and 9.7% as other Campylobacter spp. One isolate (1.6%) was positive for both C. jejuni and C. coli specific PCR. Antimicrobial susceptibility testing using the disk diffusion assay and the broth microdilution method showed resistance to: ampicillin (63% and 94.1%), ciprofloxacin (9.3% and 11.8%), erythromycin (53.7% and 70.6%), gentamicin (0% and 15.7%), streptomycin (35.2% and 84.3%), and tetracycline (18.5% and 17.7%), respectively. Resistance to azithromycin (42.6%), nalidixic acid (64.8%), and chloramphenicol (13%) was determined using the disk diffusion assay only, while resistance to tylosin (90.2%) was quantified using the broth microdilution method. The blaOXA-61 (52.6% and 28.1%), cmeB (26.3% and 31.3%), tet(O) (26.3% and 31.3%), and aph-3-1 (5.3% and 3.0%) were detected in C. coli and C. jejuni. These findings highlight the extent of antimicrobial resistance in Campylobacter occurring in important foods in Tanzania. The potential risks to consumers emphasize the need for adequate control approaches, including the prudent use of antimicrobials to minimize the spread of antimicrobial-resistant Campylobacter. PMID:26153978
Aims: Several bacteriocins (BCNs) identified from chicken commensal bacteria dramatically reduced Campylobacter colonization in poultry and aredirected toward on farm control of this important food-borne human pathogen. BCN resistance in C. jejuni is very difficult to develop in vitro. In this study...
Inglis, G D; Morck, D W; McAllister, T A; Entz, T; Olson, M E; Yanke, L J; Read, R R
Antimicrobial resistance (AMR) was temporally assessed in campylobacters isolated from beef cattle (7,738 fecal samples from 2,622 animals) in four commercial feedlots in Alberta. All calves were administered chlortetracycline and oxytetracycline in feed, and a majority of the animals (93%) were injected with long-acting oxytetracycline upon arrival at the feedlot. Fecal samples from individual animals were collected upon arrival (i.e., entry sample), 69 days (standard deviation [SD] = 3 days) after arrival (i.e., interim sample), and 189 days (SD = 33 days) after arrival (i.e., exit sample) at the feedlot. In total, 1,586 Campylobacter isolates consisting of Campylobacter coli (n = 154), Campylobacter fetus (n = 994), Campylobacter jejuni (n = 431), Campylobacter hyointestinalis (n = 4), and Campylobacter lanienae (n = 3) were recovered and characterized. The administration of antimicrobials did not decrease carriage rates of campylobacters, and minimal resistance (< or =4%) to azithromycin, ciprofloxacin, enrofloxacin, gentamicin, and meropenem was observed. In contrast, substantive increases in the prevalence of isolates resistant to tetracycline and doxycycline (56 to 89%) for C. coli, C. fetus, and C. jejuni, as well as in the number of animals (7 to 42%) from which resistant isolates were recovered, were observed during the feedlot period. Increased resistance to erythromycin (total isolates and carriages rates) was also observed in isolates of C. coli over the three isolation times. The majority of C. fetus isolates recovered were resistant to nalidixic acid, but this was independent of when they were isolated. A relatively limited number of multidrug-resistant isolates were recovered and consisted primarily of C. coli resistant to tetracyclines and erythromycin (10% of isolates). Over the course of the feedlot period, considerable increases in antimicrobial resistance were observed in C. coli, C. fetus, and C. jejuni, but with the exception of erythromycin
Campylobacter spp. are human, foodborne, and bacterial pathogens that are frequently isolated from live poultry and processed poultry products. These pathogens are classified as microaerophiles; therefore, Campylobacter cultures are generally grown in atmospheres with reduced oxygen levels and elev...
Aarestrup, F M; Nielsen, E M; Madsen, M; Engberg, J
The MICs of 16 antimicrobial agents were determined for 202 Campylobacter jejuni isolates, 123 Campylobacter coli isolates, and 6 Campylobacter lari isolates from humans and food animals in Denmark. The C. jejuni isolates originated from humans (75), broilers (95), cattle (29), and pigs (3); the C. coli isolates originated from humans (7), broilers (17), and pigs (99); and the C. lari isolates originated from broilers (5) and cattle (1). All isolates were susceptible to apramycin, neomycin, and gentamicin. Only a few C. jejuni isolates were resistant to one or more antimicrobial agents. Resistance to tetracycline was more common among C. jejuni isolates from humans (11%) than among C. jejuni isolates from animals (0 to 2%). More resistance to streptomycin was found among C. jejuni isolates from cattle (10%) than among those from humans (4%) or broilers (1%). A greater proportion of C. coli than of C. jejuni isolates were resistant to the other antimicrobial agents tested. Isolates were in most cases either coresistant to tylosin, spiramycin, and erythromycin or susceptible to all three antibiotics. More macrolide-resistant isolates were observed among C. coli isolates from swine (79%) than among C. coli isolates from broilers (18%) and humans (14%). Twenty-four percent of C. coli isolates from pigs were resistant to enrofloxacin, whereas 29% of C. coli isolates from humans and none from broilers were resistant. More resistance to streptomycin was observed among C. coli isolates from swine (48%) than among C. coli isolates from broilers (6%) or humans (0%). The six C. lari isolates were susceptible to all antimicrobial agents except ampicillin and nalidixic acid. This study showed that antimicrobial resistance was found only at relatively low frequencies among C. jejuni and C. lari isolates. Among C. coli isolates, especially from swine, there was a high level of resistance to macrolides and streptomycin. Furthermore, this study showed differences in the resistance
Yee, Emma; Huynh, Stephen; Chapman, Mary H.; Parker, Craig T.
Campylobacter iguaniorum is a member of the C. fetus group of campylobacters and is one of two Campylobacter taxa isolated from reptiles. This study describes the whole-genome sequence of the C. iguaniorum strain RM11343, which was isolated from a California alpaca fecal sample. PMID:27365359
Campylobacter iguaniorum is a member of the C. fetus group of campylobacters and is one of two Campylobacter taxa isolated from reptiles. This study describes the whole-genome sequence of the C. iguaniorum strain RM11343, which was isolated from a California alpaca fecal sample....
Campylobacter iguaniorum is a member of the C. fetus group of campylobacters and is one of two Campylobacter taxa isolated from reptiles. This study describes the whole-genome sequence of the C. iguaniorum strain RM11343, which was isolated from a California alpaca fecal sample....
Gaudreau, Christiane; Boucher, France; Gilbert, Huguette; Bekal, Sadjia
From 2002 to 2013 in Montreal, Quebec, Canada, 38 Campylobacter coli isolates were more frequently erythromycin, tetracycline, and ciprofloxacin resistant than 440 Campylobacter jejuni subsp. jejuni isolates (18.4% versus 1.8%; P = 0.00005), of which the 148 isolates acquired abroad were more frequently erythromycin, tetracycline, and ciprofloxacin resistant than the 292 isolates acquired locally (5.4% versus 0%; P = 0.0001).
Campylobacter iguaniorum has been isolated from reptiles. This Campylobacter species is genetically related to C. fetus and C. hyointestinalis. Here we present the first whole genome sequence for this species....
Lee, K; Iwata, T; Nakadai, A; Kato, T; Hayama, S; Taniguchi, T; Hayashidani, H
To estimate the public and animal health risk that alien species pose, the prevalence of Salmonella, Yersinia, and Campylobacter spp. in feral raccoons (Procyon lotor, n=459) and masked palm civets (Paguma larvata, n=153), which are abundant alien species in Japan, was investigated in urban and suburban areas of Japan. Salmonella enterica was detected from 29 samples [26 raccoons, 5.7%, 95% confidence interval (CI) 7.8-3.5%; three masked palm civets, 2.0%, 95% CI 4.2-0%]. Many of the isolates belonged to serovars that are commonly isolated from human gastroenteritis patients (e.g. S. Infantis, S. Typhimurium, and S. Thompson). The antimicrobial susceptibility test showed that 26.9 % of the isolates from raccoons were resistant to at least one antimicrobial agent, whereas none of the isolates from masked palm civets were resistant. Yersinia sp. was detected from 193 samples (177 raccoons, 38.6%, 95% CI 43.0-34.1%; 16 masked palm civets, 10.5%, 95% CI 15.3-5.6%). All virulent Yersinia strains belonged to Yersinia pseudotuberculosis, which was isolated from seven (1.5%, 95% CI 2.6-0.4%) raccoons and six (3.9%, 95% CI 7.0-0.8%) masked palm civets. According to the detection of virulence factors, all the Y. pseudotuberculosis isolates belonged to the Far Eastern systemic pathogenicity type. Campylobacter spp. was detected from 17 samples (six raccoons, 1.3%, 95% CI 2.3-0.3%; 11 masked palm civets, 7.2%, 95% CI 11.3-3.1%). Among these, three isolates from raccoons were identified as C. jejuni. These results showed that these pathogens can be transmitted by human activities, other wild animals, and the environment to feral raccoons and masked palm civets, and vice versa. As these animals have omnivorous behaviour and a wide range of habitats, they can play an important role in the transmission of the enteric pathogens.
During independent diagnostic screenings of otariid seals in California (US) and phocid seals in Scotland (UK), Campylobacter-like isolates, which differed from the established Campylobacter taxa, were cultured from abscesses and internal organs of different seal species. A polyphasic study was unde...
Introduction: Campylobacter spp. are recognized as important agents of human foodborne gastroenteritis. To monitor trends in food safety and public health, antimicrobial susceptibility testing of Campylobacter derived from poultry products and infected patients has become common practice in both r...
Noormohamed, Aneesa; Fakhr, Mohamed K
Studies that investigate arsenic resistance in the foodborne bacterium Campylobacter are limited. A total of 552 Campylobacter isolates (281 Campylobacter jejuni and 271 Campylobacter coli) isolated from retail meat samples were subjected to arsenic resistance profiling using the following arsenic compounds: arsanilic acid (4-2,048 μg/mL), roxarsone (4-2048 μg/mL), arsenate (16-8,192 μg/mL) and arsenite (4-2,048 μg/mL). A total of 223 of these isolates (114 Campylobacter jejuni and 109 Campylobacter coli) were further analyzed for the presence of five arsenic resistance genes (arsP, arsR, arsC, acr3, and arsB) by PCR. Most of the 552 Campylobacter isolates were able to survive at higher concentrations of arsanilic acid (512-2,048 μg/mL), roxarsone (512-2,048 μg/mL), and arsenate (128-1,024 μg/mL), but at lower concentrations for arsenite (4-16 μg/mL). Ninety seven percent of the isolates tested by PCR showed the presence of arsP and arsR genes. While 95% of the Campylobacter coli isolates contained a larger arsenic resistance operon that has all of the four genes (arsP, arsR, arsC and acr3), 85% of the Campylobacter jejuni isolates carried the short operon (arsP, and arsR). The presence of arsC and acr3 did not significantly increase arsenic resistance with the exception of conferring resistance to higher concentrations of arsenate to some Campylobacter isolates. arsB was prevalent in 98% of the tested Campylobacter jejuni isolates, regardless of the presence or absence of arsC and acr3, but was completely absent in Campylobacter coli. To our knowledge, this is the first study to determine arsenic resistance and the prevalence of arsenic resistance genes in such a large number of Campylobacter isolates.
Van Hoang, Ky; Stern, Norman J.; Lin, Jun
Aims Several bacteriocins (BCNs) that were identified from chicken commensal bacteria dramatically reduced Campylobacter colonization in poultry and are being directed toward on-farm control of this important foodborne human pathogen. A recent study has shown that BCN resistance in C. jejuni is very difficult to develop in vitro. In this study, in vivo development and stability of BCN resistance in Campylobacter was examined. Methods and Results Chickens infected with C. jejuni NCTC 11168 were treated with BCN E-760 at the dose of 5 mg/kg body weight/day via oral gavages for three consecutive days, which selected BCN-resistant (BCNr) mutants in the treated birds. However, all the in vivo-selected mutants only displayed low-levels of resistance to BCN (MIC = 2–8 mg/L) when compared to parent strain (MIC = 0.5 mg/L). Inactivation of CmeABC efflux pump of the BCNr mutants led to increased susceptibility to BCN (8–32 fold MIC reduction). Three different BCNr Campylobacter strains (in vitro- or in vivo-derived) were examined for the stability of BCN resistance using both in vitro and in vivo systems. The low-level of BCN resistance in these strains was not stable in vitro or in vivo in the absence of BCN selection pressure. Conclusions Usage of BCN E-760 only selected low-level BCNr C. jejuni mutants in vivo and the low-level BCN resistance was not stable in vitro and in vivo. Significance and Impact of the Study The study provides helpful information for risk assessment of the future practical application of the anti-Campylobacter BCNs in animals. PMID:21973216
Hiett, Kelli L
Currently, there is no universally accepted standard media or method for the recovery of Campylobacter species. This is likely due to the ubiquity of the organism in nature, the complex sample matrices from which the organism is often recovered, as well as the fragile/viable-but nonculturable state the organism assumes in response to stress. The use of a sterile filter placed upon a nonselective Brucella Agar Blood Plate (BAB), followed by incubation at 37 °C in a hydrogen-containing atmosphere (Campycheck), is one method to recover stressed and emerging Campylobacter spp. from complex environmental matrices; however, this technique does not currently allow for the enumeration of the recovered organisms. Enumeration is performed using serial dilutions of sample homogenate plated onto modified Campy-Cefex media followed by incubation at either 37 °C or 42 °C in a microaerobic atmosphere.
Bratz, Katharina; Gölz, Greta; Janczyk, Pawel; Nöckler, Karsten; Alter, Thomas
Campylobacter (C.) spp. are well recognised as the leading cause of bacterial food-borne diarrheal disease worldwide, with C. jejuni and C. coli as the most important species: C. coli is highly abundant in pigs and pork meat has often been implicated as a source for human infection. Intestinal colonisation of C. coli in pigs plays a role in carcass contamination during slaughter. Different pre-harvest intervention measures are proposed to reduce the C. coli burden in the porcine intestine. Among others, the use of probiotics to prevent or reduce the colonisation of intestinal pathogens is discussed. One aim of this study was to screen a variety of probiotics to evaluate their inhibitory activity against Campylobacter spp. in vitro. Therefore, cell-free culture supernatants of Lactobacillus spp., Bifidobacterium spp., Enterococcus (E.) faecium NCIMB 10415, and Escherichia coli Nissle 1917 were tested against C. jejuni and C. coli by a well-diffusion agar assay. Seven out of eleven Lactobacillus strains showed an inhibitory activity against at least one of the three tested Campylobacter strains. This antagonistic activity against Campylobacter spp. was caused by the production of organic acids that lowered the pH. Application with pH neutralised cell-free culture supernatants abolished this inhibitory effect. Other tested strains with probiotic properties showed no inhibitory activity against any Campylobacter spp. strain. The strain E. faecium NCIMB 10415 was chosen to test its inhibitory activity against C. coli in vivo. Twenty weaned piglets were allocated into two groups, a probiotic group and a control group.The diet of the probiotic group was supplemented with E. faecium NCIMB 10415 (10(9) cfu/kg feed, Cylactin) since weaning, whereas the control group received no probiotic treatment. All piglets were naturally colonised with C. coli. The excretion load of C. coli was monitored for 28 days. The results indicate that dietary supplementation of E. faecium NCIMB
Moore, J E; Caldwell, P S; Millar, B C; Murphy, P G
The occurrence of Campylobacter spp was examined in a variety of waters in Northern Ireland. Conventional cultural techniques were employed with 768 water specimens, including drinking waters (tap, spring, bore hole and bottled) and recreational waters (swimming pool, lough, river and sea). Positive waters included 1/11 (9.1%) drinking water from untreated well water, as well as 5/12 (41.7%) untreated surface waters from loughs and 7/8 (87.5%) untreated river waters. Overall, untreated surface waters may represent a source of contamination with Campylobacter spp. in Northern Ireland, where they have a recreational involvement or are used as a drinking source by man or agricultural livestock. Therefore waterborne campylobacteriosis should be considered in patients presenting with acute enteritis and a history of participation in water sports/activities. As faecal coliform organisms have been previously shown to be poor markers of water quality, especially for Campylobacter spp, new criteria should be established to assess the risk of this infection and to evaluate and monitor the quality of water used for recreational purposes.
de Boer, P; Rahaoui, H; Leer, R J; Montijn, R C; van der Vossen, J M B M
The major disadvantage of the current gold standard for detection of the food pathogen Campylobacter, i.e. culturing, is the lengthy procedure. In this study we assessed the use of real-time PCR for detection of Campylobacter. To this end, 926 poultry samples, taken from transport containers and broiler caeca in The Netherlands in 2007, were subjected to three different real-time PCR detection methods: one targeting the Campylobacter jejuni hipO gene, one targeting the Campylobacter coli glyA gene, and one generically targeting Campylobacter spp. 16S rDNA sequence. The PCR results from the three different PCR protocols were compared to the work of Nauta et al. (2009) who analyzed the same set of samples collected from 62 broiler flocks by means of enrichment culturing. The results indicate that the generic 16S campylobacter PCR detection is equally reliable but much faster (4 h instead of ≥2 days) than detection by means of culturing. Moreover, PCR detection targeting the hipO and the glyA gene provide the possibility of C. jejuni and C. coli species discrimination. The generic Campylobacter spp. PCR analysis also confirmed the high incidence of Campylobacter spp. in poultry samples (∼90%) and the species specific PCR showed the simultaneous presence of C. jejuni and C. coli in ∼24% of the samples. Furthermore, the results from the three PCR analyses suggested the occurrence of alternative Campylobacter species in almost 10% of the samples. The campylobacter PCR detection methods reported here can replace traditional culturing because of being quicker and more reliable.
Sampers, Imca; Habib, Ihab; De Zutter, Lieven; Dumoulin, Ann; Uyttendaele, Mieke
The survival of Campylobacter spp. under defined conditions of freezing (-22 degrees C) was studied in naturally contaminated chicken skin and minced chicken meat. A decline of approximately one log(10) cfu/g was observed after 1 day of freezing. No further significant reduction was achieved by prolonged storage in the freezer, although a tendency for further gradual reduction of the numbers of Campylobacter spp. present was noted. Campylobacter spp. could still be detected qualitatively (per 0.1g) after 84 days. In a second part of this study, the survival of Campylobacter spp. in a typical minced meat preparation (minced meat supplemented with 1.5% salt (NaCl)) stored at refrigeration (4 degrees C) or frozen (-22 degrees C) was studied. No significant reduction of the pathogen was observed if the minced chicken meat was kept at 4 degrees C for 14 days, opposite to approximately one log(10) cfu/g reduction after 1 day when the minced meat preparation was stored in the freezer (-22 degrees C) for 14 days. The latter reduction is imputed to the effect of freezing as mentioned above and not due to the supplementation of NaCl to minced meat or the combination of NaCl and freezing, because similar reductions of Campylobacter spp. were noticed when minced meat (without addition of NaCl) was frozen. Finally, in a third part of the study, the survival of Campylobacter spp. subjected to a heat treatment, conform to consumer-based pan-frying, in inoculated (4.5+/-0.2 cfu/g) as well as naturally contaminated chicken burgers (2.1+/-0.1 cfu/g) was studied. The Campylobacter spp. numbers declined after 2 min (internal temperature reached circa 38 degrees C), where after 4 min (internal temperature reached circa 57.5 degrees C) they dropped below detectable levels (<10 cfu/g).
Wei, Bai; Cha, Se-Yeoun; Kang, Min; Roh, Jae-Hee; Seo, Hye-Suk; Yoon, Ran-Hee; Jang, Hyung-Kwan
Campylobacter is a food-borne zoonotic pathogen that causes human gastroenteritis worldwide. Campylobacter bacteria are commensal in the intestines of many food production animals, including ducks and chickens. The objective of the study was to determine the prevalence of Campylobacter species in domestic ducks, and the agar dilution method was used to determine resistance of the isolates to eight antibiotics. In addition, multilocus sequence typing (MLST) was performed to determine the sequence types (STs) of selected Campylobacter isolates. Between May and September 2012, 58 duck farms were analyzed, and 56 (96.6%) were positive for Campylobacter. Among the isolates, 82.1% were Campylobacter jejuni, 16.1% were C. coli, and one was unidentified by PCR. Of the 46 C. jejuni isolates, 87.0%, 10.9%, and 21.7% were resistant to ciprofloxacin, erythromycin, and azithromycin, respectively. Among the C. coli isolates, all 9 strains were resistant to ampicillin, and 77.8% and 33.3% were resistant to ciprofloxacin and azithromycin, respectively. The majority of the Campylobacter isolates were classified as multidrug resistant. Twenty-eight STs were identified, including 20 STs for C. jejuni and 8 STs for C. coli. The most common clonal complexes in C. jejuni were the ST-21 complex and the ST-45 complex, while the ST-828 complex predominated in C. coli. The majority of isolates were of STs noted in ducks and humans from earlier studies, along with seven STs previously associated only with human disease. These STs overlapped between duck and human isolates, indicating that Campylobacter isolates from ducks should be considered potential sources of human infection.
Fennell, C L; Totten, P A; Quinn, T C; Patton, D L; Holmes, K K; Stamm, W E
Thirteen Campylobacter-like organisms (CLOs) isolated from rectal cultures from homosexual men were studied. Like catalase-positive Campylobacter species, CLOs were curved gram-negative rods that did not grow aerobically, were motile, were oxidase- and catalase-positive, and did not utilize glucose. However, CLOs could not be classified within any of the Campylobacter species because they grew slowly and had unusual colony morphology; did not grow at 25 C, hydrolyze hippurate, produce H2S in triple sugar-iron agar, or tolerate 2% NaCl; were inhibited by 30-micrograms disks of nalidixic acid; and tolerated 1% glycine and 0.04% triphenyltetrazolium chloride. Three groups of CLOs were identified based on differences in nitrate reduction, growth at 42 C, and sensitivity to cephalothin. By the colony hybridization technique, whole-cell DNA isolated from a strain in each CLO group hybridized with DNA from other strains in the same group, but not with strains in other groups or with reference strains of catalase-positive Campylobacter species.
Nielsen, Hans Linde; Engberg, Jørgen; Ejlertsen, Tove; Nielsen, Henrik
One thousand seven hundred ninety-one diarrheic stool samples were cultivated for Campylobacter spp. We found a high prevalence of Campylobacter concisus with use of a polycarbonate filter (n = 114) compared to a cellulose acetate filter (n = 79) (P < .0001). The polycarbonate filter is superior to the commonly used cellulose acetate filter for detection of C. concisus.
Murinda, S E; Nguyen, L T; Nam, H M; Almeida, R A; Headrick, S J; Oliver, S P
Six visits were conducted to four dairy farms to collect swab, liquid, and solid dairy farm environmental samples (165 to 180/farm; 15 sample types). The objective of the study was to determine on-farm sources of Campylobacter jejuni, Salmonella spp., Listeria monocytogenes, and Shiga toxin-producing Escherichia coli (STEC), which might serve as reservoirs for transmission of pathogens. Samples were analyzed using mostly U.S. Food and Drug Administration's Bacteriological Analytical Manual protocols; however, Salmonella spp., L. monocytogenes and STEC were co-enriched in universal pre-enrichment broth. Campylobacter jejuni were enriched in Bolton broth containing Bolton broth supplement. Pathogens were isolated on agar media, typed biochemically, and confirmed using multiplex polymerase chain reaction protocols. Campylobacter jejuni, Salmonella spp., L. monocytogenes, Sorbitol-negative (SN)-STEC O157:H7, and sorbitol-positive (SP)-STEC, respectively, were isolated from 5.06%, 3.76%, 6.51%, 0.72%, and 17.3% of samples evaluated. Whereas other pathogens were isolated from all four farms, SN-STEC O157:H7 were isolated from only two farms. Diverse serotypes of SP-STEC including O157:H7, O26:H11, O111, and O103 were isolated. None of the five pathogen groups studied were isolated from bulk tank milk (BTM). Most pathogens (44.2%) were isolated directly from fecal samples. Bovine fecal samples, lagoon water, bedding, bird droppings, and rat intestinal contents constituted areas of major concern on dairy farms. Although in-line milk filters from two farms tested positive for Salmonella or L. monocytogenes, none of the pathogens were detected in the corresponding BTM samples. Good manure management practices, including control of feral animals, are critical in assuring dairy farm hygiene. Identification of on-farm pathogen reservoirs could aid with implementation of farm-specific pathogen reduction programs.
Lehner, Y; Reich, F; Klein, G
Campylobacter spp. are the most important food-borne pathogens in broilers. Exposure of the consumer can be influenced by the reduction of contaminated broiler meat at various steps along the production line. This study was performed at a poultry slaughterhouse in Germany. Steps within the slaughter process were defined by the slaughterhouse quality control for potential Campylobacter reduction. Their impact was tested for two process variations. The first process variation was the increase of the temperature of the scalding water from 53.0 to 53.9 °C. The second step was the application of an additional outside sprayer which was placed after plucking. The increase of the scalding water temperature was the most effective measure (>2 log reduction), but resulted in defects to the broiler skin. This would limit marketing of fresh broiler meat with skin. The additional water spray after plucking had no additional effect. In fact, numbers of Campylobacter were lower before introduction of the sprayer. In conclusion, modifications of the processing technology have to be evaluated carefully, but can have additional effects for Campylobacter reduction.
Messelhäusser, Ute; Thärigen, Diana; Fella, Christiane; Schreiner, Hermann; Busch, Ulrich; Höller, Christiane
Thermotolerant Campylobacter spp. rank among the most important foodborne pathogens in Germany. Therefore a necessity for rapid and routinely useable detection methods exists also in the area of food microbiology. A reliable, cultura qualitative, but also quantitative detection of thermotolerant Campylobacter spp. pose a challenge, at least concerning special food matrices, especially because in the context of official food control the cultural detection of thermotolerant Campylobacter spp. is needed. This was the reason, why different cultural detection methods, beside the standard procedure of ISO 10272:2006, in combination with molecular and immunological screening methods were tested at the Bavarian Health and Food Safety Authority (LGL) during the last years for the use in routine diagnostic using different food matrices of animal and plant origin. The results of the comparative studies showed clearly that no enrichment broth tested gave completely satisfactory results for an only culture-based detection the combination with a screening method is therefore recommended for a rapid and reliable detection. But in this case the user should take into account that the sensitivity of such molecular and immunological methods is normally so high that in some cases, depending on the food matrix and processing step, the isolation of the pathogen would not be possible in samples, which were positive in the screening methods.
Bertasi, Barbara; Losio, Marina Nadia; Daminelli, Paolo; Finazzi, Guido; Serraino, Andrea; Piva, Silvia; Giacometti, Federica; Massella, Elisa; Ostanello, Fabio
In temperate climates, a seasonal trend was observed in the incidence of human campylobacteriosis cases, with peaks reported in spring and autumn in some countries, or in summer in others; a similar trend was observed in Campylobacter spp. dairy cattle faecal shedding, suggesting that cattle may play a role in the seasonal peak of human infection. The objectives of this study were to assess if a seasonal trend in thermophilic Campylobacter spp. contamination of raw milk exists and to evaluate a possible relation between this and the increase of human campylobacteriosis incidence in summer months. The results showed a mean prevalence of 1.6% of milk samples positive for thermophilic Campylobacter spp. with a wide range (0.0-3.1%) in different months during the three years considered. The statistical analysis showed a significant difference (P<0.01) of the prevalence of positive samples for thermophilic Campylobacter spp. between warmer and cooler months (2.3 vs 0.6%). The evidence of a seasonal trend in thermophilic Campylobacter spp. contamination of raw milk sold for direct consumption, with an increase of the prevalence in warmer months, may represent one of the possible links between seasonal trend in cattle faecal shedding and seasonal trend in human campylobacteriosis. PMID:27853714
Bertasi, Barbara; Losio, Marina Nadia; Daminelli, Paolo; Finazzi, Guido; Serraino, Andrea; Piva, Silvia; Giacometti, Federica; Massella, Elisa; Ostanello, Fabio
In temperate climates, a seasonal trend was observed in the incidence of human campylobacteriosis cases, with peaks reported in spring and autumn in some countries, or in summer in others; a similar trend was observed in Campylobacter spp. dairy cattle faecal shedding, suggesting that cattle may play a role in the seasonal peak of human infection. The objectives of this study were to assess if a seasonal trend in thermophilic Campylobacter spp. contamination of raw milk exists and to evaluate a possible relation between this and the increase of human campylobacteriosis incidence in summer months. The results showed a mean prevalence of 1.6% of milk samples positive for thermophilic Campylobacter spp. with a wide range (0.0-3.1%) in different months during the three years considered. The statistical analysis showed a significant difference (P<0.01) of the prevalence of positive samples for thermophilic Campylobacter spp. between warmer and cooler months (2.3 vs 0.6%). The evidence of a seasonal trend in thermophilic Campylobacter spp. contamination of raw milk sold for direct consumption, with an increase of the prevalence in warmer months, may represent one of the possible links between seasonal trend in cattle faecal shedding and seasonal trend in human campylobacteriosis.
Havelaar, A. H.; de Wit, M. A.; van Koningsveld, R.; van Kempen, E.
Infection with thermophilic Campylobacter spp. usually leads to an episode of acute gastroenteritis. Occasionally, more severe diseases may be induced, notably Guillain Barré syndrome and reactive arthritis. For some, the disease may be fatal. We have integrated available data in one public health measure, the Disability Adjusted Life Year (DALY). DALYs are the sum of Years of Life Lost by premature mortality and Years Lived with Disability, weighted with a factor between 0 and 1 for the severity of illness. The mean health burden of campylobacter-associated illness in the Dutch population in the period 1990-5 is estimated as 1400 (90% CI 900-2000) DALY per year. The main determinants of health burden are acute gastroenteritis (440 DALY), gastroenteritis related mortality (310 DALY) and residual symptoms of Guillain-Barré syndrome (340 DALY). Sensitivity analysis demonstrated that alternative model assumptions produced results in the above-mentioned range. PMID:11218201
Rosengren, Leigh B; Waldner, Cheryl L; Reid-Smith, Richard J; Valdivieso-Garcia, Alfonso
Campylobacter spp. (n = 405), isolated from the feces of apparently healthy grow-finish pigs in 20 herds, were tested for susceptibility to 10 antimicrobials representing seven classes. Twelve percent of the isolates were susceptible to all drugs, while 64% were resistant to two or more antimicrobial classes. Resistance was most common to clindamycin, azithromycin, and erythromycin (71% each), and 10% of the isolates were resistant to ciprofloxacin. An antimicrobial use risk-factor analysis and a variance analysis explored the connection between antimicrobial resistance and the herd. The antimicrobial exposure of each production phase of each herd, through feed and water, was evaluated as a potential risk factor for resistance to macrolides and quinolones. Every 100,000 pig days of macrolide exposure in nursery pigs increased the odds of resistance to macrolides by a factor of 1.3. In contrast, the odds of resistance to a quinolone were nine times higher in Campylobacter from herds without beta-lactam exposure in grow-finish pigs compared with those with exposure. The variance analysis identified remarkably high clustering between isolates within herds; the intraclass correlations for resistances ranged from 0.52 to 0.82. Such extreme clustering demonstrates the potential for herd-level interventions to influence antimicrobial resistance in Campylobacter. The three key findings of this study, i.e., the prevalent resistance to macrolides, the association between macrolide exposure and Campylobacter resistance to macrolides, and the high clustering of resistance within herds, illustrate the need for continued study of antimicrobial-resistant Campylobacter on pig farms and the importance of judicious antimicrobial use in pork production.
Fontanot, Marco; Iacumin, Lucilla; Cecchini, Francesca; Comi, Giuseppe; Manzano, Marisa
The detection of Campylobacter, the most commonly reported cause of foodborne gastroenteritis in the European Union, is very important for human health. The most commonly recognised risk factor for infection is the handling and/or consumption of undercooked poultry meat. The methods typically applied to evaluate the presence/absence of Campylobacter in food samples are direct plating and/or enrichment culture based on the Horizontal Method for Detection and Enumeration of Campylobacter spp. (ISO 10272-1B: 2006) and PCR. Molecular methods also allow for the detection of cells that are viable but cannot be cultivated on agar media and that decrease the time required for species identification. The current study proposes the use of two molecular methods for species identification: dot blot and PCR. The dot blot method had a sensitivity of 25 ng for detection of DNA extracted from a pure culture using a digoxigenin-labelled probe for hybridisation; the target DNA was extracted from the enrichment broth at 24 h. PCR was performed using a pair of sensitive and specific primers for the detection of Campylobacter jejuni and Campylobacter coli after 24 h of enrichment in Preston broth. The initial samples were contaminated by 5 × 10 C. jejuni cells/g and 1.5 × 10(2)C. coli cells/g, thus the number of cells present in the enrichment broth at 0 h was 1 or 3 cell/g, respectively.
Vaz, C S L; Voss-Rech, D; Pozza, J S; Coldebella, A; Silva, V S
Conventional culturing methods enable the detection of Campylobacter in broiler flocks. However, laboratory culture of Campylobacter is laborious because of its fastidious behavior and the presence of competing nontarget bacteria. This study evaluated different protocols to isolate Campylobacter from broiler litter, feces, and cloacal and drag swabs. Samples taken from commercial Brazilian broiler flocks were directly streaked onto Preston agar (PA), Campy-Line agar (CLA), and modified charcoal cefoperazone deoxycholate agar (mCCDA) and also enriched in blood-free Bolton broth (bfBB) for 24 and 48 h followed by plating onto the different selective media. Higher numbers of Campylobacter-positive cloacal and drag swab samples were observed using either direct plating or enrichment for 24 h before plating onto PA, compared with enrichment for 48 h (P < 0.05). Furthermore, direct plating was a more sensitive method to detect Campylobacter in broiler litter and feces samples. Analysis of directly plated samples revealed that higher Campylobacter levels were detected in feces streaked onto PA (88.8%), cloacal swabs plated onto mCCDA (72.2%), drag swabs streaked onto CLA or mCCDA (69.4%), and litter samples inoculated onto PA (63.8%). Preston agar was the best agar to isolate Campylobacter from directly plated litter samples (P < 0.05), but there was no difference in the efficacies of PA, mCCDA, and CLA in detecting Campylobacter in other samples. The isolated Campylobacter strains were phenotypically identified as Campylobacter jejuni or Campylobacter coli. The predominant contaminant observed in the Campylobacter cultures was Proteus mirabilis, which was resistant to the majority of antimicrobial agents in selective media. Together, these data showed that direct plating onto PA and onto either CLA or mCCDA as the second selective agar enabled the reliable isolation of thermophilic Campylobacter species from broiler samples. Finally, Campylobacter was detected in all
Evangelopoulou, Grammato; Filioussis, Georgios; Kritas, Spyridon; Kantere, Maria; Burriel, Angeliki R
The presence of Gram-negative bacteria species, other than Salmonella spp., in the gallbladder of pigs was examined. Isolated Gram-negative bacteria were assigned to species using the Microgen™ GnA+B-ID Systems. Of the 64 isolated strains 43 were identified as Escherichia coli, seven as Enterobacter spp., three each as Klebsiella spp., Citrobacterfreundii, Aeromonas hydrophila and Cronobacter sakazakii and one each as Escherichiafergusonii and Trabulsiella guamensis. Their antibiograms showed very high resistance to ampicillin, amoxicillin, tetracycline, chloramphenicol and sulfamethoxazole/trimethoprim. It was concluded that the pigs' gallbladder is a reservoir of potentially pathogenic Gram-negative bacteria for pork consumers.
Evans, Mary C; Wegener, Henrik C
The use of antimicrobial growth promoters in Danish food animal production was discontinued in 1998. Contrary to concerns that pathogen load would increase; we found a significant decrease in Salmonella in broilers, swine, pork, and chicken meat and no change in the prevalence of Campylobacter in broilers.
Dhamabutra, N; Kamol-Rathanakul, P; Pienthaweechai, K
The 100 ml of canal water samples of 36 canals in Bangkok Metropolitan Area were examined in three periods starting from July-September 1988, November 1988-January 1989 and February-April 1989. Each time the 52 water samples were checked. Of 156 water samples, 116 strains of Campylobacter species were isolated. They were 63.79 per cent (74 strains) of C. cryaerophila and 36.21 per cent (42 strains) of C. cryaerophila-like organisms. The differentiation was determined by urease activity test. C. cryaerophila was isolated from 44.23 per cent (23 strains), 51.19 per cent (27 strains) and 46.15 per cent (24 strains) and also C. cryaerophila-like organism from 28.85 per cent (15 strains), 19.23 per cent (10 strains) and 32.69 per cent (17 strains) of the 52 samples during each period respectively. Since C. cryaerophila and C. cryaerophila-like are aerotolerant Campylobacter, they grow well in aerobic conditions at 25 degrees-36 degrees C. On the contrary, thermophilic Campylobacter such as C. jejuni, C. coli and C. laridis require atmosphere containing 5 per cent O2, 10 per cent CO2, 85 per cent N2 and temperature at 36 degrees-42 degrees C, so the environment in the canals is unfavorable for their growth. The etiological role of C. cryaerophila in pathogenesis in humans is still unknown, and requires furthers study. This study shows that canals can be an important source of these two Campylobacter species that might be potential pathogens in the future.
Nathues, C; Grüning, P; Fruth, A; Verspohl, J; Blaha, T; Kreienbrock, L; Merle, R
Campylobacter spp., Salmonella enterica, and Yersinia enterocolitica are common causes of foodborne infections in humans with pork as a potential source. Monitoring programs at farm level are, to date, only implemented for S. enterica, while epidemiological knowledge of the other two pathogens is still lacking. This study aimed to assess the pathogen load (in the pigs' environment) in fattening pig herds, their simultaneous occurrence, and the occurrence of Campylobacter spp. and Y. enterocolitica in herds in different Salmonella risk categories. In 50 fattening pig herds in northern Germany, four pooled fecal samples and 10 swab samples from the pigs' direct environment (pen walls, nipple drinkers), indirect environment (hallways, drive boards), and flies and rodent droppings were collected from each herd and submitted for cultural examination. Campylobacter spp. were detected in 38.1% of fecal, 32.7% of direct environment, 5.3% of indirect environment, and 4.6% of flies/pests samples collected, and Y. enterocolitica in 17.1, 8.1, 1.2, and 3.1% and S. enterica in 11.2, 7.7, 4.1, and 1.5%, respectively. For Campylobacter spp., Y. enterocolitica, and S. enterica, 80, 48, and 32% of herds were positive, respectively; 22 herds were positive for both Campylobacter spp. and Y. enterocolitica, 12 for Campylobacter spp. and S. enterica, and 7 for Y. enterocolitica and S. enterica. There was no significant association between the pathogens at herd level. Campylobacter spp. and Y. enterocolitica were found more often in samples from the low Salmonella risk category (odds ratio, 0.51; confidence interval, 0.36 to 0.73, and 0.3, 0.17 to 0.57), and this was also the case for Y. enterocolitica at herd level (odds ratio, 0.08; confidence interval, 0.02 to 0.3). This study provides evidence that the pigs' environment should be accounted for when implementing control measures on farms against Campylobacter spp. and Y. enterocolitica. An extrapolation from the current Salmonella
Schweitzer, Nóra; Dán, Ádám; Kaszanyitzky, Éva; Samu, Péterné; Tóth, Ádám György; Varga, János; Damjanova, Ivelina
Campylobacter spp. are the most common cause of bacterial enteritis in Hungary, and the aim of this study was to identify the distribution, genotypes, and antimicrobial susceptibility of Campylobacter species in the most important food-producing animals at the time of slaughter during 2008 and 2009. Of 1,110 samples, 266 were identified as Campylobacter coli (23.9%) and 143 as C. jejuni (12.9%) by real-time PCR. Resistance to enrofloxacin-ciprofloxacin and nalidixic acid was significant, especially in C. jejuni (73.3%) and C. coli (77.2%) from broilers. Higher erythromycin (P = 0.043) and tetracycline (P = 1.865e-14) resistance rates were found among C. coli isolates (9.7 and 74.1%, respectively) than among C. jejuni isolates (3.1 and 36.6%, respectively). A total of 47 fla short variable region sequences were identified among 73 selected C. coli and C. jejuni isolates, with 35 fla types detected only once. At the nucleotide level, fla types A66 and A21 were the most common. Using the pulsed-field gel electrophoresis method, 66% of strains exhibited unique profiles after Sma I digestion. Forty-two isolates assigned to 18 Sma I clusters were further typed by Kpn I, and of these, 24 were assigned to 10 Kpn I clusters. For isolates in five Kpn I clusters, epidemiological links were observed. Stable C. jejuni and C. coli clones were detected, indicating that further studies involving broiler and human isolates need to be conducted to elucidate the importance of these stable clones in human infections.
van de Giessen, A. W.; Tilburg, J. J.; Ritmeester, W. S.; van der Plas, J.
Transmission routes of Campylobacter spp. in broilers and possibilities for prevention of infections were studied on two Dutch broiler farms. The occurrence of Campylobacter spp. was studied in successive broiler flocks, in the environment of the farms and in some of the parent flocks involved. Isolates of Campylobacter spp. were typed by using randomly amplified polymorphic DNA (RAPD) analysis. The results indicate that broiler flocks become infected from environmental sources. The typing results suggest that on one farm transmission of Campylobacter spp. occurred from cattle to broilers via the farmer's footwear. After several campylobacter positive broiler cycles hygiene measures, including thorough cleaning and disinfection procedures, change of footwear at the entrance of each broiler house, control of vermin and other hygienic precautions, were introduced on both farms in order to prevent transmission of Campylobacter spp. from the farm environment to the broilers. The results indicate that the application of hygiene measures significantly reduced campylobacter infections of broiler flocks on both farms. PMID:9747756
subtyping Salmonella spp ., Shigella spp ., and Vibrio spp ., in addition to C. jejttni 112,13]. Unlike other enteric bacteria, Campylobacter is a...characterizing C. jejuni and C. coli and identifying specific Campylobacter spp . in outbreak studies [14-16]. Furthermore, the combination of PFGE...ibilities for Campylobacter spp . [32-35]. However, good agreement of antimicrobial susceptibility test between disk diffusion and agar dilution tests has
Gabriele-Rivet, Vanessa; Fairbrother, Julie-Hélène; Tremblay, Donald; Harel, Josée; Côté, Nathalie; Arsenault, Julie
Feral pigeons (Columbia livia) can harbor a range of zoonotic pathogens. A transversal study was undertaken to estimate the prevalence of feral pigeons infected by various pathogens in public areas in Montreal, Quebec. Cloacal swabs from captured birds were cultured for Salmonella spp. and Campylobacter spp. and tested by real-time polymerase chain reaction (RT-PCR) for the detection of Coxiella burnetii. An oropharyngeal swab was also submitted to real-time reverse-transcription polymerase chain reaction (RRT-PCR) for the detection of Newcastle disease virus. Among the 187 pigeons tested from 10 public areas, 9.1% (95% CI: 3.0 to 15.2) were positive for Campylobacter spp. with all strains identified as Campylobacter jejuni. The Campylobacter status of birds was not associated with individual characteristics of birds, with the exception of body score. None of the pigeons tested positive for the other pathogens. Direct or indirect contacts with feral pigeons may constitute a potential risk for Campylobacter infection in humans. PMID:26733736
Gabriele-Rivet, Vanessa; Fairbrother, Julie-Hélène; Tremblay, Donald; Harel, Josée; Côté, Nathalie; Arsenault, Julie
Feral pigeons (Columbia livia) can harbor a range of zoonotic pathogens. A transversal study was undertaken to estimate the prevalence of feral pigeons infected by various pathogens in public areas in Montreal, Quebec. Cloacal swabs from captured birds were cultured for Salmonella spp. and Campylobacter spp. and tested by real-time polymerase chain reaction (RT-PCR) for the detection of Coxiella burnetii. An oropharyngeal swab was also submitted to real-time reverse-transcription polymerase chain reaction (RRT-PCR) for the detection of Newcastle disease virus. Among the 187 pigeons tested from 10 public areas, 9.1% (95% CI: 3.0 to 15.2) were positive for Campylobacter spp. with all strains identified as Campylobacter jejuni. The Campylobacter status of birds was not associated with individual characteristics of birds, with the exception of body score. None of the pigeons tested positive for the other pathogens. Direct or indirect contacts with feral pigeons may constitute a potential risk for Campylobacter infection in humans.
Smith, Shaun; Meade, Joseph; McGill, Kevina; Gibbons, James; Bolton, Declan; Whyte, Paul
Extended spectrum β-lactamase (ESBL) producing Escherichia coli have emerged as a contaminant on modified charcoal cefoperazone deoxycholate agar (mCCDA) when attempting to selectively isolate Campylobacter spp. from poultry. E. coli are particularly problematic given their ability to grow under microaerophilic conditions and have been shown to outcompete Campylobacter species making Campylobacter detection or enumeration difficult. This paper recommends a novel method for restoring the selectivity of mCCDA using tazobactam, a β-lactamase inhibitor. The method significantly inhibited ESBL E. coli growth in spiked or naturally contaminated broiler caecal samples (p≤0.01) when compared to conventional mCCDA. This effect was seen at concentrations as low as 1mg/L tazobactam. TmCCDA(1) was found to inhibit up to 8 log10 CFU/mL of ESBL E. coli in mixed pure cultures and 7.5 log10 CFU/mL in caecal samples. Furthermore TmCCDA concentrations up to 10 mg/L had no statistically significant inhibitory effect (p≥0.05) on the recovery of a panel of 27 Campylobacter jejuni and 5 Campylobacter coli isolates when compared to conventional mCCDA. From this study it is suggested that tazobactam, which is more chemically stable than clavulanic acid or sulbactam, is more suitable for restoring the selectivity of mCCDA for the detection or isolation of campylobacters.
A polyphasic study was undertaken to determine the taxonomic position of 13 Campylobacter fetus-like isolates from humans (n=8) and reptiles (n=5). Phenotypic characterization, Genusgenus-specific and sap insertion-PCR initially identified all human isolates as type A Campylobacter fetus. Phylogenet...
Kaur, Taranjit; Singh, Jatinder; Huffman, Michael A; Petrzelková, Klára J; Taylor, Nancy S; Xu, Shilu; Dewhirst, Floyd E; Paster, Bruce J; Debruyne, Lies; Vandamme, Peter; Fox, James G
The transmission of simian immunodeficiency and Ebola viruses to humans in recent years has heightened awareness of the public health significance of zoonotic diseases of primate origin, particularly from chimpanzees. In this study, we analyzed 71 fecal samples collected from 2 different wild chimpanzee (Pan troglodytes) populations with different histories in relation to their proximity to humans. Campylobacter spp. were detected by culture in 19/56 (34%) group 1 (human habituated for research and tourism purposes at Mahale Mountains National Park) and 0/15 (0%) group 2 (not human habituated but propagated from an introduced population released from captivity over 30 years ago at Rubondo Island National Park) chimpanzees, respectively. Using 16S rRNA gene sequencing, all isolates were virtually identical (at most a single base difference), and the chimpanzee isolates were most closely related to Campylobacter helveticus and Campylobacter upsaliensis (94.7% and 95.9% similarity, respectively). Whole-cell protein profiling, amplified fragment length polymorphism analysis of genomic DNA, hsp60 sequence analysis, and determination of the mol% G+C content revealed two subgroups among the chimpanzee isolates. DNA-DNA hybridization experiments confirmed that both subgroups represented distinct genomic species. In the absence of differential biochemical characteristics and morphology and identical 16S rRNA gene sequences, we propose to classify all isolates into a single novel nomenspecies, Campylobacter troglodytis, with strain MIT 05-9149 as the type strain; strain MIT 05-9157 is suggested as the reference strain for the second C. troglodytis genomovar. Further studies are required to determine whether the organism is pathogenic to chimpanzees and whether this novel Campylobacter colonizes humans and causes enteric disease.
To speciate Campylobacter strains from the ceca of chickens in Grenada by PCR and to evaluate DNA-based typing methods for the characterization of these isolates. Isolates were speciated with two multiplex PCR assays and were typed with flaA-RFLP, PFGE and MLST. Results confirmed that C. coli strain...
Cornelius, A J; Nicol, C; Hudson, J A
Sheep liver samples were tested for the presence and numbers of Campylobacter jejuni and C. coli during both spring and autumn. Over the same period, isolates were obtained from human clinical cases from the same geographical area as where the food samples were purchased. A subset of the C. jejuni isolates was typed by both Penner serotyping and pulsed field gel electrophoresis using the restriction enzyme SmaI, to estimate the proportion of liver isolate types that were also isolated from human cases of campylobacteriosis. Of the 272 liver samples tested, 180 (66.2%) contained Campylobacter. Most of the positive samples contained <3 MPN/g of the organism, and only 12 (6.7%) were contaminated at a level exceeding 100 MPN/g. A total of 180 C. jejuni isolates were obtained from sheep liver and another 200 from human faeces. Of these, 212 isolates were randomly selected for typing, half from raw liver and half from human faeces. More than half (61.1%) of the 106 C. jejuni isolates from liver were of subtypes that were also isolated from human cases. While the C. jejuni present in sheep liver were mostly of subtypes also isolated from human cases, the significance of this food as a vehicle of human campylobacteriosis needs to be examined further in respect to other factors such as dose-response information, consumption data, frequency of undercooking and cross contamination.
CABALLERO, Moisés; RIVERA, Isabel; JARA, Luis M.; ULLOA-STANOJLOVIC, Francisco M.; SHIVA, Carlos
SUMMARY Feral pigeons (Columbia livia) live in close contact with humans and other animals. They can transmit potentially pathogenic and zoonotic agents. The objective of this study was to isolate and detect strains of diarrheagenic Escherichia coli and Campylobacter jejuni of urban feral pigeons from an area of Lima, Peru. Fresh dropping samples from urban parks were collected for microbiological isolation of E. coli strains in selective agar, and Campylobacter by filtration method. Molecular identification of diarrheagenic pathotypes of E.coli and Campylobacter jejuni was performed by PCR. Twenty-two parks were sampled and 16 colonies of Campylobacter spp. were isolated. The 100% of isolates were identified as Campylobacter jejuni. Furthermore, 102 colonies of E. coliwere isolated and the 5.88% resulted as Enteropathogenic (EPEC) type and 0.98% as Shiga toxin-producing E. coli (STEC). The urban feral pigeons of Lima in Peru can act as a reservoir or carriers of zoonotic potentially pathogenic enteric agents. PMID:26603225
Agunos, Agnes; Waddell, Lisa; Léger, David; Taboada, Eduardo
Campylobacter and antimicrobial-resistant Campylobacter are frequently isolated from broiler chickens worldwide. In Canada, campylobacteriosis is the third leading cause of enteric disease and the regional emergence of ciprofloxacin-resistant Campylobacter in broiler chickens has raised a public health concern. This study aimed to identify, critically appraise, and synthesize literature on sources of Campylobacter in broilers at the farm level using systematic review methodology. Literature searches were conducted in January 2012 and included electronic searches in four bibliographic databases. Relevant studies in French or English (n = 95) conducted worldwide in any year and all study designs were included. Risk of Bias and GRADE criteria endorsed by the Cochrane collaboration was used to assess the internal validity of the study and overall confidence in the meta-analysis. The categories for on-farm sources were: broiler breeders/vertical transfer (number of studies = 32), animals (n = 57), humans (n = 26), environment (n = 54), and water (n = 63). Only three studies examined the antimicrobial resistance profiles of Campylobacter from these on-farm sources. Subgroups of data by source and outcome were analyzed using random effect meta-analysis. The highest risk for contaminating a new flock appears to be a contaminated barn environment due to insufficient cleaning and disinfection, insufficient downtime, and the presence of an adjacent broiler flock. Effective biosecurity enhancements from physical barriers to restricting human movement on the farm are recommended for consideration to enhance local on-farm food safety programs. Improved sampling procedures and standardized laboratory testing are needed for comparability across studies. Knowledge gaps that should be addressed include farm-level drug use and antimicrobial resistance information, further evaluation of the potential for vertical transfer, and improved genotyping methods to
Behringer, Megan; Miller, William G; Oyarzabal, Omar A
We analyzed 100 Campylobacter spp. isolates (C. jejuni and C. coli) from Grenada, Puerto Rico and Alabama, which were collected from live broilers or retail broiler meat. We analyzed these isolates with four molecular typing methods: restriction fragment length polymorphism of the flaA gene (flaA-RFLP), multilocus sequence typing (MLST), pulsed-field gel electrophoresis (PFGE), and automated repetitive extragenic palindromic polymerase chain reaction (REP-PCR) using the DiversiLab system. All methods performed similarly for the typing of C. jejuni and C. coli. The DNA extraction method appears to influence the results obtained with REP-PCR. This method was better for the typing of C. jejuni than C. coli, however both REP-PCR and flaA-RFLP generated types that were indistinguishable between C. jejuni and C. coli and appeared to be random, without any relationship to species, location, or source of isolates. PFGE and MLST generated typing results that had a better correlation with the geographic location of the isolates and showed higher concordance with the Wallace coefficient. The adjusted Rand coefficient did not show higher concordance among the methods, although the PFGE/MLST combination exhibited the highest concordance. PFGE and MLST revealed a better discriminatory power for C. coli isolates than REP-PCR or flaA-RFLP. The use of readily available online tools to calculate the confidence interval of the Simpson's index of diversity and the adjusted Rand and Wallace coefficients helped estimate the discriminatory power of typing methods. Further studies using different C. jejuni and C. coli strains may expand our understanding of the benefits and limitations of each of these typing methods for epidemiological studies of Campylobacter spp.
On, S L
A strain from human diarrhea originally identified as Campylobacter mucosalis (NCTC 12408) was examined by using 64 phenotypic characters. The similarity of this strain to 297 isolates of Campylobacter, Helicobacter, Arcobacter, and related taxa was then determined with a computer-supported data analysis program, MVSP. NCTC 12408 showed closest similarity to 20 type, reference, and field isolates of Campylobacter concisus. These strains were clearly separated from those of C. mucosalis in the numerical analysis of phenotypic tests; a table was constructed from the data used to aid in differentiating these two species in the clinical laboratory. The identity of NCTC 12408 was confirmed as C. concisus by visual comparison of its sodium dodecyl sulfate-polyacrylamide gel whole-cell protein electrophoregram with those of type strains of C. concisus and C. mucosalis. These data suggest that genuine human infection with C. mucosalis has not yet been reported.
Stable concentrated emulsions of the 1-monoglyceride of capric acid (monocaprin) with microbicidal activities against the food-borne bacteria Campylobacter jejuni, Salmonella spp., and Escherichia coli.
Thormar, Halldor; Hilmarsson, Hilmar; Bergsson, Gudmundur
Of 11 fatty acids and monoglycerides tested against Campylobacter jejuni, the 1-monoglyceride of capric acid (monocaprin) was the most active in killing the bacterium. Various monocaprin-in-water emulsions were prepared which were stable after storage at room temperature for many months and which retained their microbicidal activity. A procedure was developed to manufacture up to 500 ml of 200 mM preconcentrated emulsions of monocaprin in tap water. The concentrates were clear and remained stable for at least 12 months. They were active against C. jejuni upon 160- to 200-fold dilution in tap water and caused a >6- to 7-log(10) reduction in viable bacterial count in 1 min at room temperature. The addition of 0.8% Tween 40 to the concentrates as an emulsifying agent did not change the microbicidal activity. Emulsions of monocaprin killed a variety of Campylobacter isolates from humans and poultry and also killed strains of Campylobacter coli and Campylobacter lari, indicating a broad anticampylobacter activity. Emulsions of 1.25 mM monocaprin in citrate-lactate buffer at pH 4 to 5 caused a >6- to 7-log(10) reduction in viable bacterial counts of Salmonella spp. and Escherichia coli in 10 min. C. jejuni was also more susceptible to monocaprin emulsions at low pH. The addition of 5 and 10 mM monocaprin emulsions to Campylobacter-spiked chicken feed significantly reduced the bacterial contamination. These results are discussed in view of the possible utilization of monocaprin emulsions in controlling the spread of food-borne bacteria from poultry to humans.
Sirirak, T; Voravuthikunchai, S P
The antibacterial activity of ethanolic extracts of selected Thai medicinal plants (Rhodomyrtus tomentosa (Aiton) Hassk., Quercus infectoria G. Olivier, and Eleutherine americana Merr.) against Campylobacter spp. was investigated. Sixty-five Campylobacter, including 39 isolates from humans and 26 isolates from chicken samples, were tested. Reference Campylobacter spp. that are commonly encountered in gastroenteritis were included. The ethanolic extract of E. americana demonstrated good antibacterial activity against all the tested isolates. Inhibition zones ranged from 10 to 37 mm. Minimum inhibitory concentration (MIC) of the extract against Campylobacter isolates from humans and chicken samples ranged from 31.25 to 500 μg/mL and 62.50 to 1,000 μg/mL, respectively. The minimum bactericidal concentration ranged from 31.25 to 1,000 μg/mL for isolates from humans and 125 to 1,000 μg/mL from chicken isolates. The bactericidal activity of the ethanolic extracts of E. americana against important Campylobacter spp., including Campylobacter coli MUMT 18630, Campylobacter fetus ATCC 27374, Campylobacter jejuni ATCC 81176, Campylobacter lari ATCC 43675, and Campylobacter upsaliensis DMST 19055, were assessed at MIC, 2 MIC, and 4 MIC by counting viable cells after various time intervals. At 4 MIC, the level of the tested isolates decreased by 2 to 5 log-fold within 8 h. The ethanolic extract of E. americana demonstrated antibacterial activity against all Campylobacter spp. from both human and chicken isolates. Further investigation of this plant species may provide an alternative medicine for Campylobacter infection and an effective food additive to prevent the infection.
A cross-sectional study examining Campylobacter and other zoonotic enteric pathogens in dogs that frequent dog parks in three cities in south-western Ontario and risk factors for shedding of Campylobacter spp.
Procter, T D; Pearl, D L; Finley, R L; Leonard, E K; Janecko, N; Reid-Smith, R J; Weese, J S; Peregrine, A S; Sargeant, J M
An estimated 6 million pet dogs live in Canadian households with the potential to transmit zoonotic pathogens to humans. Dogs have been identified as carriers of Salmonella, Giardia and Campylobacter spp., particularly Campylobacter upsaliensis, but little is known about the prevalence and risk factors for these pathogens in pet dogs that visit dog parks. This study examined the prevalence of these organisms in the faeces of dogs visiting dog parks in three cities in south-western Ontario, as well as risk factors for shedding Campylobacter spp. and C. upsaliensis. From May to August 2009, canine faecal samples were collected at ten dog parks in the cities of Guelph and Kitchener-Waterloo, Ontario, Canada. Owners were asked to complete a questionnaire related to pet characteristics and management factors including age, diet and activities in which the dog participates. Faecal samples were collected from 251 dogs, and 189 questionnaires were completed. Salmonella, Giardia and Campylobacter spp. were present in 1.2%, 6.4% and 43.0% of faecal samples, respectively. Of the Campylobacter spp. detected, 86.1% were C. upsaliensis, 13% were C. jejuni and 0.9% were C. coli. Statistically significant sparing factors associated with the shedding of Campylobacter spp. included the feeding of a commercial dry diet and the dog's exposure to compost. Age of dog had a quadratic effect, with young dogs and senior dogs having an increased probability of shedding Campylobacter spp. compared with adult dogs. The only statistically significant risk factor for shedding C. upsaliensis was outdoor water access including lakes and ditches, while dogs >1 year old were at a lower risk than young dogs. Understanding the pet-related risk factors for Campylobacter spp. and C. upsaliensis shedding in dogs may help in the development of awareness and management strategies to potentially reduce the risk of transmitting this pathogen from dogs to humans.
Esson, Diane; Mather, Alison E.; Scanlan, Eoin; Gupta, Srishti; de Vries, Stefan P. W.; Bailey, David; Harris, Simon R.; McKinley, Trevelyan J.; Méric, Guillaume; Berry, Sophia K.; Mastroeni, Pietro; Sheppard, Samuel K.; Christie, Graham; Thomson, Nicholas R.; Parkhill, Julian; Maskell, Duncan J.; Grant, Andrew J.
Campylobacter jejuni, the most common cause of bacterial diarrhoeal disease, is normally helical. However, it can also adopt straight rod, elongated helical and coccoid forms. Studying how helical morphology is generated, and how it switches between its different forms, is an important objective for understanding this pathogen. Here, we aimed to determine the genetic factors involved in generating the helical shape of Campylobacter. A C. jejuni transposon (Tn) mutant library was screened for non-helical mutants with inconsistent results. Whole genome sequence variation and morphological trends within this Tn library, and in various C. jejuni wild type strains, were compared and correlated to detect genomic elements associated with helical and rod morphologies. All rod-shaped C. jejuni Tn mutants and all rod-shaped laboratory, clinical and environmental C. jejuni and Campylobacter coli contained genetic changes within the pgp1 or pgp2 genes, which encode peptidoglycan modifying enzymes. We therefore confirm the importance of Pgp1 and Pgp2 in the maintenance of helical shape and extended this to a wide range of C. jejuni and C. coli isolates. Genome sequence analysis revealed variation in the sequence and length of homopolymeric tracts found within these genes, providing a potential mechanism of phase variation of cell shape. PMID:27910897
Hidano, Arata; Yamamoto, Takehisa; Hayama, Yoko; Muroga, Norihiko; Kobayashi, Sota; Nishida, Takeshi; Tsutsui, Toshiyuki
A rapid increase in antimicrobial resistance in Campylobacter has been posing a serious concern for human health. In this study, we aimed to demonstrate the overall trend in antimicrobial resistance among Campylobacter isolates obtained from chicken meat and offal products collected from a wide geographic area throughout Japan. Resistance to Enrofloxacin was most frequently observed, with significantly higher rate of resistance among isolates obtained from offal (55.6%) than from meat (27.3%) samples (p = 0.05). These results highlight need for a better understanding of the characteristics of Campylobacter isolates obtained from chicken meat and offal products.
Campylobacter jejuni is the leading foodborne pathogen that causes human acute bacterial gastroenteritis worldwide. Human cases have been linked to consumption and/or handling of contaminated poultry products. Although Campylobacter jejuni is commonly regarded as a commensal in broiler cecal micro...
Bonetta, Si; Pignata, C; Lorenzi, E; De Ceglia, M; Meucci, L; Bonetta, Sa; Gilli, G; Carraro, E
The aim of this study was the evaluation of the occurrence of pathogenic Campylobacter, Escherichia coli O157:H7, E. coli virulence genes and Salmonella spp. in different wastewater treatment plants (WWTPs) using a method based on an enrichment step and PCR. This method was sensitive enough to detect low levels (∼2 CFU100 ml(-1) of raw sewage) of all the investigated pathogens. In the WWTP samples, E. coli O157:H7 DNA and the eae gene were never found, but 33 % of influents and effluents exhibited amplicons corresponding to Shiga-like toxin I. Twenty-five percent of the influent and 8 % of the effluent exhibited the presence of Shiga-like toxin II. Campylobacter jejuni and C. coli DNA were identified in 50 and 25 % of the influents and in 8 and 25 % of the effluents, respectively. Salmonella spp. DNA was present in all the samples. Considering the results obtained, the method tested here offers a reliable and expeditious tool for evaluating the efficiency of the effluent treatment in order to mitigate contamination risk. Influent contamination by Salmonella spp. and Campylobacter spp. provides indirect information about their circulation; moreover, their presence in effluents underlines the role of WWTPs in the contamination of the receiving surface waters, which affects public health directly or indirectly.
Tambalo, Dinah D; Boa, Tyler; Aryal, Bijaya; Yost, Christopher K
Campylobacter spp. are a substantial cause of gastroenteritis worldwide. Human infection can result from ingestion of contaminated food or water from a variety of sources, including the consumption of fresh produce that is contaminated with the pathogen via the use of contaminated irrigation water. Using molecular methods, we investigated the occurrence of Campylobacter in the Qu'Appelle River watershed, an important source of irrigation water for vegetable producers in southern Saskatchewan, Canada. Water samples were collected from 7 sampling sites from April to September 2009 (145 samples), and from 5 sampling sites from May to October 2013 (116 samples). Campylobacter was detected in 57% and 16% of the samples collected in 2009 and 2013, respectively. Campylobacter detection was highest in May and June for both sampling years. In 2009, the predominant species were Campylobacter lari and Campylobacter jejuni, with prevalences of 84% and 41%, respectively. Other Campylobacter spp. were detected less frequently. Only C. lari was detected in 2013. The results in 2009 demonstrate the species richness of Campylobacter in water sources within the watershed. The occurrence of Campylobacter in the study area also underscores the importance of monitoring irrigation water used to irrigate fresh produce from a public health prospective.
Samosornsuk, Worada; Asakura, Masahiro; Yoshida, Emi; Taguchi, Takashi; Eampokalap, Bunchuay; Chaicumpa, Wanpen; Yamasaki, Shinji
Campylobacter-induced diarrhea is increasingly recognized worldwide. However, little information is available regarding the Campylobacter strains associated with diarrheal patients in Thailand. In this study, we attempted to isolate Campylobacter strains from diarrheal patients in Thailand and to characterize the species using a cytolethal distending toxin (cdt) gene-based C. jejuni, C. coli, and C. fetus-specific multiplex PCR assay. Campylobacter species were also confirmed using 16S rRNA gene sequencing and hipO gene detection. From 2,500 diarrheal stool specimens, 76 Campylobacter-like organisms were isolated and identified via conventional culture methods. Among these 76 organisms, 73 were identified as Campylobacter species (43 C. jejuni, 29 C. coli, and 1 C. fetus) via multiplex PCR, whereas 3 remained unidentified. Two Campylobacter-like organisms yielded 2 amplicons corresponding to cdt genes from C. jejuni and C. coli. Subsequently, C. jejuni and C. coli were reisolated from each sample. The third isolate was identified as C. hyointestinalis via 16S rRNA gene sequencing. To our knowledge, this is the first report on the isolation of C. hyointestinalis from a diarrheal patient in Thailand. These data indicate that C. jejuni (58%) and C. coli (40%) are prevalent among diarrheal patients in Thailand.
Sifré, Elodie; Salha, Ben Amor; Ducournau, Astrid; Floch, Pauline; Chardon, Hubert; Mégraud, Francis; Lehours, Philippe
Antimicrobial susceptibility testing of Campylobacter isolates is of great importance for treatment options especially in systemic diseases. The European Committee for Antimicrobial Susceptibility Testing (EUCAST) recently proposed epidemiological cut-offs (ECOFFs) for a limited number of antimicrobial compounds and for Campylobacter jejuni and Campylobacter coli only. In the present study, the EUCAST method was used after minor modifications to define antimicrobial susceptibility patterns for, 1997 C. jejuni, 419 C. coli and 100 Campylobacter fetus strains received at the French National Reference Center for Campylobacters and Helicobacters. Our results show that the ECOFFs defined by EUCAST for tetracycline and ciprofloxacin can be used for C. jejuni and C. coli. The same ECOFF can be used for erythromycin for the three species. The C. jejuni and C. coli ECOFFs for ciprofloxacin however cannot be applied to C. fetus. We also provide data to categorise two 2 β-lactams of interest for systemic diseases, ampicillin and amoxicillin+clavulanate, for the three species.
We developed a biphasic method for recovery and enumeration of campylobacters. The biphasic system was composed of 96-well microtiter plates containing Campy-Line Agar (CLA) prepared 2X for all ingredients except for 1X agar (0.1 ml per well). Samples of pure Campylobacter spp. isolates, post-chi...
Valdivieso-Garcia, Alfonso; Harris, Kathleen; Riche, Edward; Campbell, Stephanie; Jarvie, Anne; Popa, Maria; Deckert, Anne; Reid-Smith, Richard; Rahn, Kris
Culture procedures for isolation of thermophilic campylobacters from food matrices are complex, labor intensive, and time-consuming. Most available methods include the use of antibiotics as selective agents to prevent the growth of competing microflora. A simple procedure for isolation of thermophilic campylobacters after enrichment in Rosef's enrichment broth was developed using a hydrophobic grid membrane filter (HGMF) on semisolid medium (SSM). SSM contains no antibiotics, and the HGMF physically separates Campylobacter from the enrichment broth, allowing isolation based on differential motility. The HGMF-SSM method was compared to the Agriculture and Agri-Food Canada Food Safety Procedures Manual (FSPM-10) method (Isolation of Thermophilic Campylobacters from Fresh Pork, Beef Veal, Poultry and Ready-to-Eat Meat Products), which includes the use of selective antibiotics. During the initial study, after enrichment the HGMF-SSM method yielded pure cultures of campylobacters after 16 to 18 h (overnight) compared with 48 h for the FSPM-10 method. Ninety-four turkey samples collected at local retail stores and 38 frozen pig fecal samples were processed by both methods. Thirty-five samples (26.5%) were positive by the HGMF-SSM method; 24 (18.2%) of these positive samples contained Campylobacter jejuni and 11 (8.3%) contained Campylobacter coli. With the FSPM-10 method, 25 samples (18.9%) were positive: 21 (15.9%) with C. jejuni and 4 (3%) with C. coli. For a subsequent field study, only the HGMF-SSM method was used to isolate Campylobacter from 1,200 chicken samples and 454 turkey samples sold at retail. Analysis of five subisolates from various samples indicated that only one type of Campylobacter was recovered by the HGMF-SSM method, as ascertained by MICs for 10 antimicrobials, sequencing of the short variable region of the flaA gene, and fingerprinting based on amplified fragment length polymorphism. The absence of antibiotics in the SSM may explain the higher
Ryu, Hodon; Vogel, Jason; Santo Domingo, Jorge; Ashbolt, Nicholas J.
The risk to human health of the annual sandhill crane (Grus canadensis) migration through Nebraska, which is thought to be a major source of fecal pollution of the central Platte River, is unknown. To better understand potential risks, the presence of Campylobacter species and three fecal indicator bacterial groups (Enterococcus spp., Escherichia coli, and Bacteroidetes) was assayed by PCR from crane excreta and water samples collected during their stopover at the Platte River, Nebraska, in 2010. Genus-specific PCR assays and sequence analyses identified Campylobacter jejuni as the predominant Campylobacter species in sandhill crane excreta. Campylobacter spp. were detected in 48% of crane excreta, 24% of water samples, and 11% of sediment samples. The estimated densities of Enterococcus spp. were highest in excreta samples (mean, 4.6 × 108 cell equivalents [CE]/g), while water samples contained higher levels of Bacteroidetes (mean, 5.1 × 105 CE/100 ml). Enterococcus spp., E. coli, and Campylobacter spp. were significantly increased in river water and sediments during the crane migration period, with Enterococcus sp. densities (∼3.3 × 105 CE/g) 2 to 4 orders of magnitude higher than those of Bacteroidetes (4.9 × 103 CE/g), E. coli (2.2 × 103 CE/g), and Campylobacter spp. (37 CE/g). Sequencing data for the 16S rRNA gene and Campylobacter species-specific PCR assays indicated that C. jejuni was the major Campylobacter species present in water, sediments, and crane excreta. Overall, migration appeared to result in a significant, but temporary, change in water quality in spring, when there may be a C. jejuni health hazard associated with water and crops visited by the migrating birds. PMID:23584775
Taylor, N M; Wales, A D; Ridley, A M; Davies, R H
Data on husbandry practices, performance, disease and drug use were collected during a cross-sectional survey of 89 poultry meat farms in England and Wales to provide information on possible risk factors for the occurrence of fluoroquinolone (FQ)-resistant bacteria. Faeces samples were used to classify farms as "affected" or "not affected" by FQ-resistant (FQr) Escherichia coli or Campylobacter spp. Risk factor analysis identified the use of FQ on the farms as having by far the strongest association, among the factors considered, with the occurrence of FQr bacteria. Resistant E. coli and/or Campylobacter spp. were found on 86% of the farms with a history of FQ use. However, a substantial proportion of farms with no history of FQ use also yielded FQr organisms, suggesting that resistant bacteria may transfer between farms. Further analysis suggested that for Campylobacter spp., on-farm hygiene, cleaning and disinfection between batches of birds and wildlife control were of most significance. By contrast, for E. coli biosecurity from external contamination was of particular importance, although the modelling indicated that other factors were likely to be involved. Detailed studies on a small number of sites showed that FQr E. coli can survive routine cleaning and disinfection. It appears difficult to avoid the occurrence of resistant bacteria when FQ are used on a farm, but the present findings provide evidence to support recommendations to reduce the substantial risk of the incidental acquisition of such resistance by farms where FQ are not used.
Pielsticker, C.; Glünder, G.; Rautenschlein, S.
Campylobacter is the most common bacterial food-borne pathogen worldwide. Poultry and specifically chicken and raw chicken meat is the main source for human Campylobacter infection. Whilst being colonized by Campylobacter spp. chicken in contrast to human, do scarcely develop pathological lesions. The immune mechanisms controlling Campylobacter colonization and infection in chickens are still not clear. Previous studies and our investigations indicate that the ability to colonize the chicken varies significantly not only between Campylobacter strains but also depending on the original source of the infecting isolate. The data provides circumstantial evidence that early immune mechanisms in the gut may play an important role in the fate of Campylobacter in the host. PMID:24611122
Dufour, Virginie; Alazzam, Bachar; Ermel, Gwennola; Thepaut, Marion; Rossero, Albert; Tresse, Odile; Baysse, Christine
Food-borne human infection with Campylobacter jejuni is a medical concern in both industrialized and developing countries. Efficient eradication of C. jejuni reservoirs within live animals and processed foods is limited by the development of antimicrobial resistances and by practical problems related to the use of conventional antibiotics in food processes. We have investigated the bacteriostatic and bactericidal activities of two phytochemicals, allyl-isothiocyanate (AITC), and benzyl isothiocyanate (BITC), against 24 C. jejuni isolates from chicken feces, human infections, and contaminated foods, as well as two reference strains NCTC11168 and 81-176. AITC and BITC displayed a potent antibacterial activity against C. jejuni. BITC showed a higher overall antibacterial effect (MIC of 1.25-5 μg mL(-1)) compared to AITC (MIC of 50-200 μg mL(-1)). Both compounds are bactericidal rather than bacteriostatic. The sensitivity levels of C. jejuni isolates against isothiocyanates were neither correlated with the presence of a GGT (γ-Glutamyl Transpeptidase) encoding gene in the genome, with antibiotic resistance nor with the origin of the biological sample. However the ggt mutant of C. jejuni 81-176 displayed a decreased survival rate compared to wild-type when exposed to ITC. This work determined the MIC of two ITC against a panel of C. jejuni isolates, showed that both compounds are bactericidal rather than bacteriostatic, and highlighted the role of GGT enzyme in the survival rate of C. jejuni exposed to ITC.
Price, Lance B; Johnson, Elizabeth; Vailes, Rocio; Silbergeld, Ellen
The use of fluoroquinolones (FQs) in poultry production is an important issue in public health today. In February 2002, two prominent U.S. poultry companies pledged to stop using FQs for flock-wide treatment. One year later, we began a survey of Campylobacter isolates on chicken products from these two companies and from two producers claiming total abstention from antibiotic use. Using both standard isolation methods and new methods modified to enhance detection of FQ-resistant Campylobacter, we compared rates of FQ-resistant Campylobacter among these products. Four major findings were drawn from this study: a) antibiotic-free brands were not more likely to be contaminated with Campylobacter; b) a high percentage of products from the two conventional brands were contaminated with FQ-resistant Campylobacter (43 and 96%); c) these conventional brands had significantly higher odds of carrying resistant strains compared with antibiotic-free products; and d) supplementing media with FQs increased the sensitivity of detecting FQ-resistant strains among mixed populations of Campylobacter, thus reducing a bias toward underestimating the prevalence of FQ-resistant Campylobacter on samples. These results suggest that FQ resistance may persist in the commercial poultry environment in the absence of FQ-selective pressure and that these strains contaminate a larger proportion of foods than reported previously.
Wang, Yang; Zhang, Maojun; Deng, Fengru; Shen, Zhangqi; Wu, Congming; Zhang, Jianzhong
Antibiotic-resistant Campylobacter constitutes a serious threat to public health, and resistance to macrolides is of particular concern, as this class of antibiotics is the drug of choice for clinical therapy of campylobacteriosis. Very recently, a horizontally transferrable macrolide resistance mediated by the rRNA methylase gene erm(B) was reported in a Campylobacter coli isolate, but little is known about the dissemination of erm(B) among Campylobacter isolates and the association of erm(B)-carrying isolates with clinical disease. To address this question and facilitate the control of antibiotic-resistant Campylobacter, we determined the distribution of erm(B) in 1,554 C. coli and Campylobacter jejuni isolates derived from food-producing animals and clinically confirmed human diarrheal cases. The results revealed that 58 of the examined isolates harbored erm(B) and exhibited high-level resistance to macrolides, and most were recent isolates, derived in 2011-2012. In addition, the erm(B)-positive isolates were all resistant to fluoroquinolones, another clinically important antibiotic used for treating campylobacteriosis. The erm(B) gene is found to be associated with chromosomal multidrug resistance genomic islands (MDRGIs) of Gram-positive origin or with plasmids of various sizes. All MDRGIs were transferrable to macrolide-susceptible C. jejuni by natural transformation under laboratory conditions. Molecular typing of the erm(B)-carrying isolates by pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST) identified diverse genotypes and outbreak-associated diarrheal isolates. Molecular typing also suggested zoonotic transmission of erm(B)-positive Campylobacter. These findings reveal an emerging and alarming trend of dissemination of erm(B) and MDRGIs in Campylobacter and underscore the need for heightened efforts to control their further spread. PMID:24982085
associated with significant foodborne disease. Campylobacter jejuni and C. coli are the two most prevalent species contributing to human diarrheal disease. The objective of this study was to determine the routes of transmission for Campylobacter throughout turkey production and processing. A floc...
Piccirillo, Alessandra; Niero, Giulia; Calleros, Lucía; Pérez, Ruben; Naya, Hugo; Iraola, Gregorio
During a screening study to determine the presence of species of the genus Campylobacter in reptiles, three putative strains (RC7, RC11 and RC20T) were isolated from different individuals of the western Hermann's tortoise (Testudo hermanni hermanni). Initially, these isolates were characterized as representing Campylobacterfetus subsp. fetus by multiplex PCR and partial 16S rRNA gene sequence analysis. Further whole- genome characterization revealed considerable differences compared to other Campylobacter species. A polyphasic study was then undertaken to determine the exact taxonomic position of the isolates. The three strains were characterized by conventional phenotypic tests and whole genome sequencing. We generated robust phylogenies that showed a distinct clade containing only these strains using the 16S rRNA and atpA genes and a set of 40 universal proteins. Our phylogenetic analysis demonstrates their designation as representing a novel species and this was further confirmed using whole- genome average nucleotide identity within the genus Campylobacter (~80 %). Compared to most Campylobacter species, these strains hydrolysed hippurate, and grew well at 25 °C but not at 42 °C. Phenotypic and genetic analyses demonstrate that the three Campylobacter strains isolated from the western Hermann's tortoise represent a novel species within the genus Campylobacter, for which the name Campylobactergeochelonis sp. nov. is proposed, with RC20T (=DSM 102159T=LMG 29375T) as the type strain.
Background: Foodborne illness is a serious public health problem. According to the U.S. Food and Drug Administration Campylobacter jejuni is the leading cause of bacterial diarrheal illness in the United States, causing more disease than Shigella spp. and Salmonella spp. combined. The CDC estima...
El-Zamkan, Mona A.; Hameed, Karima G. Abdel
Aim: This study was accomplished to test raw milk and certain dairy products sold in local markets of Qena, Egypt, for the presence of Campylobacter coli and Campylobacter jejuni. Materials and Methods: A total of 150 samples of raw milk, kareish cheese, and yoghurt (50 samples each) were subjected first to enrichment in Bolton broth at 42°C for 2 days under a microaerobic condition, subsequently campylobacter blood free selective agar plates were cultured and incubated in the same condition of the broth. Based on the morphological and biochemical themes of the growing colonies, it was further classified into Campylobacter spp. The identified isolates were later affirmed by polymerase chain reaction using primers that were designed to locate hipO genes in C. jejuni and glyA in C. coli. Results: Of the total 150 examined samples of raw milk and soft cheese samples; 37 (24.6%) samples were contaminated with Campylobacter spp. C. jejuni was dominating in this study in 20%, 14%, and 8% of the examined raw milk, kareish cheese, and yoghurt samples, respectively. No sample harbored C. coli. Conclusion: Campylobacter spp. could be detected in 24.6% of the investigated samples. C. jejuni isolated from 14% of the total tested samples, while C. coli could not be detected from the examined samples. Campylobacter spp. is rampant in the areas of poor hygienic conditions making products made from raw milk of public health hazard. PMID:27847427
Ramonaitė, Sigita; Novoslavskij, Aleksandr; Zakarienė, Gintarė; Aksomaitienė, Jurgita; Malakauskas, Mindaugas
The occurrence, seasonal variation and genetic diversity of Campylobacter spp. in pigeons and crows over a 1-year period were evaluated. Campylobacter spp. were isolated from 166 (34.6 %) out of 480 wild bird faecal samples. The occurrence of Campylobacter spp. in faecal samples was higher among crows (39.2 %) than pigeons (30.0 %), (P < 0.05). Campylobacter jejuni was the most common species detected among wild bird faecal samples (98.2 %). Meanwhile, Campylobacter coli prevalence in wild bird faecal samples was low-6 %. The Simpson's diversity index of C. jejuni flaA RFLP types was lower in pigeons (D = 0.88) compared with C. jejuni isolates detected in crows (D = 0.97). Obtained results revealed that C. jejuni are widely prevalent among crows and pigeons, indicating these wild birds as potential infection sources to humans. Further studies are required to determine crows and pigeons role in zoonotic transmission of Campylobacter.
González, Mario; Paz Villanueva, Maria; Debruyne, Lies; Vandamme, Peter; Fernández, Heriberto
Campylobacter insulaenigrae have been isolated from different pinnipeds but not from South American sea lion (Otaria flavescens).The aim of this work is to report the first isolation of C. insulaenigrae from South American sea lion (Otaria flavescens).The isolate, identified by its phenotypic and molecular characteristics, allow recognizing O. flavescens as a new host for C. insulaenigrae.
González, Mario; Paz Villanueva, Maria; Debruyne, Lies; Vandamme, Peter; Fernández, Heriberto
Campylobacter insulaenigrae have been isolated from different pinnipeds but not from South American sea lion (Otaria flavescens).The aim of this work is to report the first isolation of C. insulaenigrae from South American sea lion (Otaria flavescens).The isolate, identified by its phenotypic and molecular characteristics, allow recognizing O. flavescens as a new host for C. insulaenigrae. PMID:24031630
Zhao, S; Tyson, G H; Chen, Y; Li, C; Mukherjee, S; Young, S; Lam, C; Folster, J P; Whichard, J M; McDermott, P F
The objectives of this study were to identify antimicrobial resistance genotypes for Campylobacter and to evaluate the correlation between resistance phenotypes and genotypes using in vitro antimicrobial susceptibility testing and whole-genome sequencing (WGS). A total of 114 Campylobacter species isolates (82 C. coli and 32 C. jejuni) obtained from 2000 to 2013 from humans, retail meats, and cecal samples from food production animals in the United States as part of the National Antimicrobial Resistance Monitoring System were selected for study. Resistance phenotypes were determined using broth microdilution of nine antimicrobials. Genomic DNA was sequenced using the Illumina MiSeq platform, and resistance genotypes were identified using assembled WGS sequences through blastx analysis. Eighteen resistance genes, including tet(O), blaOXA-61, catA, lnu(C), aph(2″)-Ib, aph(2″)-Ic, aph(2')-If, aph(2″)-Ig, aph(2″)-Ih, aac(6')-Ie-aph(2″)-Ia, aac(6')-Ie-aph(2″)-If, aac(6')-Im, aadE, sat4, ant(6'), aad9, aph(3')-Ic, and aph(3')-IIIa, and mutations in two housekeeping genes (gyrA and 23S rRNA) were identified. There was a high degree of correlation between phenotypic resistance to a given drug and the presence of one or more corresponding resistance genes. Phenotypic and genotypic correlation was 100% for tetracycline, ciprofloxacin/nalidixic acid, and erythromycin, and correlations ranged from 95.4% to 98.7% for gentamicin, azithromycin, clindamycin, and telithromycin. All isolates were susceptible to florfenicol, and no genes associated with florfenicol resistance were detected. There was a strong correlation (99.2%) between resistance genotypes and phenotypes, suggesting that WGS is a reliable indicator of resistance to the nine antimicrobial agents assayed in this study. WGS has the potential to be a powerful tool for antimicrobial resistance surveillance programs.
Ewnetu, Desalegne; Mihret, Adane
In this study, the isolation and antimicrobial resistance of Campylobacter jejuni and Campylobacter coli strains from chickens and humans in Bahir Dar, Ethiopia, were analyzed. Two hundred and ten human and 220 chicken samples were analyzed between October 2007 and April 2008. Seventeen human and 160 chicken Campylobacter species were isolated. The overall prevalence of thermophilic campylobacters was 8% and 72.7% in humans and chickens, respectively. In humans, 94.1% of the isolates were C. jejuni and 5.9% were C. coli. C. jejuni was a predominant species of thermophilic campylobacters in all categories of patients. In chicken, 92.5% of thermophilic campylobacters isolated were C. jejuni and 7.5% were C. coli. Among the 16 isolates of C. jejuni in humans, 18.8%, 12.5%, 12.5%, 18.8%, 25%, and 22.2% were resistant to ampicillin, ciprofloxacin, erythromycin, nalidixic acid, streptomycin, and tetracycline, respectively, whereas among the 148 C. jejuni isolates from chicken, 17.5%, 14.9%, 12.2%, and 13.5% were resistant to ampicillin, erythromycin, streptomycin, and tetracycline, respectively. Among the 12 isolates of C. coli in chicken, 16.6%, 8.3%, and 16.6% were resistant to ampicillin, streptomycin, and tetracycline, respectively. The overall level of resistance was not significantly different in C. jejuni and C. coli isolates of both humans and poultry. The detection of resistant isolates for commonly used antimicrobials may cause a threat to humans and chickens by limiting therapeutic options.
Jokinen, Cassandra C; Koot, Jacqueline M; Carrillo, Catherine D; Gannon, Victor P J; Jardine, Claire M; Mutschall, Steven K; Topp, Edward; Taboada, Eduardo N
Improved isolation techniques from environmental water and animal samples are vital to understanding Campylobacter epidemiology. In this study, the efficiency of selective enrichment in Bolton Broth (BB) followed by plating on charcoal cefoperazone deoxycholate agar (CCDA) (conventional method) was compared with an approach combining BB enrichment and passive filtration (membrane method) adapted from a method previously developed for testing of broiler meat, in the isolation of thermophilic campylobacters from surface water and animal fecal samples. The conventional method led to recoveries of Campylobacter from 36.7% of the water samples and 78.0% of the fecal samples and similar numbers, 38.3% and 76.0%, respectively, were obtained with the membrane method. To investigate the genetic diversity of Campylobacter jejuni and Campylobacter coli obtained by these two methods, isolates were analyzed using Comparative Genomic Fingerprinting, a high-resolution subtyping technique. The conventional and membrane methods yielded similar numbers of Campylobacter subtypes from water (25 and 28, respectively) and fecal (15 and 17, respectively) samples. Although there was no significant difference in recovery rates between the conventional and membrane methods, a significant improvement in isolation efficiency was obtained by using the membrane method, with a false-positive rate of 1.6% compared with 30.7% obtained using the conventional method. In conclusion, although the two methods are comparable in sensitivity, the membrane method had higher specificity, making it a cost-effective procedure for the enhanced isolation of C. jejuni and C. coli from water and animal fecal samples.
Bernal, Johan F.; Donado-Godoy, Pilar; Valencia, María Fernanda; León, Maribel; Gómez, Yolanda; Rodríguez, Fernando; Agarwala, Richa; Landsman, David
Campylobacter coli, along with Campylobacter jejuni, is a major agent of gastroenteritis and acute enterocolitis in humans. We report the whole-genome sequences of two multidrug-resistance C. coli strains, isolated from the Colombian poultry chain. The isolates contain a variety of antimicrobial resistance genes for aminoglycosides, lincosamides, fluoroquinolones, and tetracycline. PMID:26988048
Bernal, Johan F; Donado-Godoy, Pilar; Valencia, María Fernanda; León, Maribel; Gómez, Yolanda; Rodríguez, Fernando; Agarwala, Richa; Landsman, David; Mariño-Ramírez, Leonardo
Campylobacter coli, along with Campylobacter jejuni, is a major agent of gastroenteritis and acute enterocolitis in humans. We report the whole-genome sequences of two multidrug-resistance C. coli strains, isolated from the Colombian poultry chain. The isolates contain a variety of antimicrobial resistance genes for aminoglycosides, lincosamides, fluoroquinolones, and tetracycline.
Giacomelli, M; Andrighetto, C; Rossi, F; Lombardi, A; Rizzotti, L; Martini, M; Piccirillo, A
Genetic variability and genotypic antimicrobial resistance (AMR) of Campylobacter jejuni and Campylobacter coli from commercial broiler farms were investigated in this study. Campylobacter isolates were genetically characterized by random amplified polymorphic DNA (RAPD)-polymerase chain reaction (PCR) and flaA-SVR and flaB-SVR sequence-based typing. Eight RAPD types were identified in C. jejuni and three in C. coli, while 16 fla profiles were detected among all isolates. Further, 13 flaA-SVR and 13 flaB-SVR alleles were identified. Both typing methods detected a high level of genetic diversity, but fla-SVR typing showed a higher discriminatory power. Indeed, Simpson's index of fla typing (D=0.920) was higher than that of RAPD typing (D=0.814). Moreover, the association of flaA-SVR and flaB-SVR sequence analysis showed a higher discriminatory power compared with the sequence analysis of single loci. Isolates were also analysed by the mismatch amplification mutation assay PCR test and the detection of cmeB gene to determine the occurrence of genetic determinants of AMR to macrolides and fluoroquinolones and multidrug resistance. The A2074C and A2075G mutations in the 23S rRNA gene, the C257T mutation in the gyrA gene, and the cmeB gene were higher in C. coli (19.0%, 67.0%, 100.0% and 100.0%, respectively) than in C. jejuni (0.0%, 3.1%, 48.3% and 48.3%, respectively). This study showed a high degree of genetic diversity and a high prevalence of genetic determinants of macrolide resistance, fluoroquinolone resistance and multidrug resistance among C. jejuni and C. coli isolates from Italian commercial broiler farms.
The emerging pathogen Campylobacter ureolyticus has been isolated from human and animal genital infections, human periodontal infections, domestic and food animals, and from cases of human gastroenteritis. We report the whole-genome sequence of the human clinical isolate RIGS 9880, which is the firs...
We examined the prevalence, quantity, and diversity of Campylobacter species in the excreta of 159 California gull samples using PCR and qPCR based detection assays. While Campylobacter prevalence and abundance was relatively high in the gull excreta examined, molecular data ind...
The unacceptably high frequency of Campylobacter jejuni transmission from poultry to humans encourages scientists to consider and create alternative intervention strategies to control the pathogen in poultry production. Extremely high numbers of Campylobacter (often >108 cfu/g of poultry intestinal...
Chandrasena, G. I.; Deletic, A.; McCarthy, D. T.
Knowledge of pathogen removal in stormwater biofilters (also known as stormwater bioretention systems or rain gardens) has predominately been determined using bacterial indicators, and the removal of reference pathogens in these systems has rarely been investigated. Furthermore, current understanding of indicator bacteria removal in these systems is largely built upon laboratory-scale work. This paper examines whether indicator organism removal from urban stormwater using biofilters in laboratory settings are representative of the removal of pathogens in field conditions, by studying the removal of Escherichia coli (a typical indicator microorganism) and Campylobacter spp. (a typical reference pathogen) from urban stormwater by two established field-scale biofilters. It was found that E. coli log reduction was higher than that of Campylobacter spp. in both biofilters, and that there was no correlation between E. coli and Campylobacter spp. log removal performance. This confirms that E. coli behaves significantly differently to this reference pathogen, reinforcing that single organisms should not be employed to understand faecal microorganism removal in urban stormwater treatment systems. The average reduction in E. coli from only one of the tested biofilters was able to meet the log reduction targets suggested in the current Australian stormwater harvesting guidelines for irrigating sports fields and golf courses. The difference in the performance of the two biofilters is likely a result of a number of design and operational factors; the most important being that the biofilter that did not meet the guidelines was tested using extremely high influent volumes and microbial concentrations, and long antecedent dry weather periods. As such, the E. coli removal performances identified in this study confirmed laboratory findings that inflow concentration and antecedent dry period impact overall microbial removal. In general, this paper emphasizes the need for the
Vidal, A B; Colles, F M; Rodgers, J D; McCarthy, N D; Davies, R H; Maiden, M C J; Clifton-Hadley, F A
The genetic diversity of Campylobacter jejuni and Campylobacter coliisolates from commercial broiler farms was examined by multilocus sequence typing (MLST), with an assessment of the impact of the sample type and laboratory method on the genotypes of Campylobacter isolated. A total of 645C. jejuniand 106C. coli isolates were obtained from 32 flocks and 17 farms, with 47 sequence types (STs) identified. The Campylobacter jejuniisolates obtained by different sampling approaches and laboratory methods were very similar, with the same STs identified at similar frequencies, and had no major effect on the genetic profile of Campylobacter population in broiler flocks at the farm level. ForC. coli, the results were more equivocal. While some STs were widely distributed within and among farms and flocks, analysis of molecular variance (AMOVA) revealed a high degree of genetic diversity among farms forC. jejuni, where farm effects accounted for 70.5% of variance, and among flocks from the same farm (9.9% of variance for C. jejuni and 64.1% forC. coli). These results show the complexity of the population structure of Campylobacterin broiler production and that commercial broiler farms provide an ecological niche for a wide diversity of genotypes. The genetic diversity of C. jejuni isolates among broiler farms should be taken into account when designing studies to understand Campylobacter populations in broiler production and the impact of interventions. We provide evidence that supports synthesis of studies on C. jejuni populations even when laboratory and sampling methods are not identical.
Tyson, G. H.; Chen, Y.; Li, C.; Mukherjee, S.; Young, S.; Lam, C.; Folster, J. P.; Whichard, J. M.; McDermott, P. F.
The objectives of this study were to identify antimicrobial resistance genotypes for Campylobacter and to evaluate the correlation between resistance phenotypes and genotypes using in vitro antimicrobial susceptibility testing and whole-genome sequencing (WGS). A total of 114 Campylobacter species isolates (82 C. coli and 32 C. jejuni) obtained from 2000 to 2013 from humans, retail meats, and cecal samples from food production animals in the United States as part of the National Antimicrobial Resistance Monitoring System were selected for study. Resistance phenotypes were determined using broth microdilution of nine antimicrobials. Genomic DNA was sequenced using the Illumina MiSeq platform, and resistance genotypes were identified using assembled WGS sequences through blastx analysis. Eighteen resistance genes, including tet(O), blaOXA-61, catA, lnu(C), aph(2″)-Ib, aph(2″)-Ic, aph(2′)-If, aph(2″)-Ig, aph(2″)-Ih, aac(6′)-Ie-aph(2″)-Ia, aac(6′)-Ie-aph(2″)-If, aac(6′)-Im, aadE, sat4, ant(6′), aad9, aph(3′)-Ic, and aph(3′)-IIIa, and mutations in two housekeeping genes (gyrA and 23S rRNA) were identified. There was a high degree of correlation between phenotypic resistance to a given drug and the presence of one or more corresponding resistance genes. Phenotypic and genotypic correlation was 100% for tetracycline, ciprofloxacin/nalidixic acid, and erythromycin, and correlations ranged from 95.4% to 98.7% for gentamicin, azithromycin, clindamycin, and telithromycin. All isolates were susceptible to florfenicol, and no genes associated with florfenicol resistance were detected. There was a strong correlation (99.2%) between resistance genotypes and phenotypes, suggesting that WGS is a reliable indicator of resistance to the nine antimicrobial agents assayed in this study. WGS has the potential to be a powerful tool for antimicrobial resistance surveillance programs. PMID:26519386
Ovesen, Sandra; Kirk, Karina Frahm; Nielsen, Hans Linde; Nielsen, Henrik
Campylobacter concisus is an emerging pathogen associated with gastrointestinal disorders such as gastroenteritis and inflammatory bowel diseases (IBD), but the species is also found in healthy subjects. The heterogeneous genome of C. concisus increases the likelihood of varying virulence between strains. Flagella motility is a crucial virulence factor for the well-recognized Campylobacter jejuni; therefore, this study aimed to analyze the motility of C. concisus isolated from saliva, gut biopsies, and feces of patients with IBD, gastroenteritis, and healthy subjects. The motility zones of 63 isolates from 52 patients were measured after microaerobic growth in soft-agar plates for 72 hours. The motility of C. concisus was significantly lower than that of Campylobacter jejuni and Campylobacter fetus subsp. fetus. The motility of C. concisus varied between isolates (4-22 mm), but there was no statistical significant difference between isolates from IBD patients and healthy subjects (p = 0.14). A tendency of a larger motility zones was observed for IBD gut mucosa isolates, although it did not reach statistical significance (p = 0.13), and no difference was found between oral or fecal isolates between groups. In conclusion, the varying motility of C. concisus could not be related to disease outcome or colonization sites.
Krueger, C L; Sheikh, W
A new medium, pseudomonas selective isolation agar, was developed to isolate Pseudomonas spp. from water. It consists of 350 micrograms of nitrofurantoin per ml and 2 micrograms of crystal violet per ml in a nutrient agar base. It is more selective for Pseudomonas spp. than are available commercial media. Its ingredients are inexpensive and readily available, and it is easy to prepare. PMID:3579287
Colles, F. M.; Rodgers, J. D.; McCarthy, N. D.; Davies, R. H.; Maiden, M. C. J.; Clifton-Hadley, F. A.
The genetic diversity of Campylobacter jejuni and Campylobacter coli isolates from commercial broiler farms was examined by multilocus sequence typing (MLST), with an assessment of the impact of the sample type and laboratory method on the genotypes of Campylobacter isolated. A total of 645 C. jejuni and 106 C. coli isolates were obtained from 32 flocks and 17 farms, with 47 sequence types (STs) identified. The Campylobacter jejuni isolates obtained by different sampling approaches and laboratory methods were very similar, with the same STs identified at similar frequencies, and had no major effect on the genetic profile of Campylobacter population in broiler flocks at the farm level. For C. coli, the results were more equivocal. While some STs were widely distributed within and among farms and flocks, analysis of molecular variance (AMOVA) revealed a high degree of genetic diversity among farms for C. jejuni, where farm effects accounted for 70.5% of variance, and among flocks from the same farm (9.9% of variance for C. jejuni and 64.1% for C. coli). These results show the complexity of the population structure of Campylobacter in broiler production and that commercial broiler farms provide an ecological niche for a wide diversity of genotypes. The genetic diversity of C. jejuni isolates among broiler farms should be taken into account when designing studies to understand Campylobacter populations in broiler production and the impact of interventions. We provide evidence that supports synthesis of studies on C. jejuni populations even when laboratory and sampling methods are not identical. PMID:26873321
Waldenström, Jonas; Broman, Tina; Carlsson, Inger; Hasselquist, Dennis; Achterberg, René P.; Wagenaar, Jaap A.; Olsen, Björn
A total of 1,794 migrating birds trapped at a coastal site in southern Sweden were sampled for detection of Campylobacter spp. All isolates phenotypically identified as Campylobacter jejuni and a subset of those identified as non-C. jejuni were identified to the species level by PCR-based techniques. C. jejuni was found in 5.0% of the birds, Campylobacter lari was found in 5.6%, and Campylobacter coli was found in 0.9%. An additional 10.7% of the tested birds were infected with hippurate hydrolysis-negative Campylobacter spp. that were not identified to the species level. The prevalence of Campylobacter spp. differed significantly between ecological guilds of birds. Shoreline-foraging birds feeding on invertebrates and opportunistic feeders were most commonly infected (76.8 and 50.0%, respectively). High prevalence was also shown in other ground-foraging guilds, i.e., ground-foraging invertebrate feeders (11.0%), ground-foraging insectivores (20.3%), and plant-eating species (18.8%). Almost no Campylobacter spp. were found in ground-foraging granivores (2.3%), arboreal insectivores (0.6%), aerial insectivores (0%), or reed- and herbaceous plant-foraging insectivores (3.5%). During the autumn migration, a high proportion of samples from juveniles were positive (7.1% in passerines, 55.0% in shorebirds), indicating transmission on the breeding grounds or during the early part of migration. Prevalence of Campylobacter spp. was associated with increasing body mass among passerine bird species. Furthermore, prevalence was higher in short-distance migrants wintering in Europe than in long-distance migrants wintering in Africa, the Middle East, or Asia. Among ground-foraging birds of the Muscicapidae, those of the subfamily Turdinae (i.e., Turdus spp.) showed a high prevalence of Campylobacter spp., while the organism was not isolated in any member of the subfamily Muscicapinae (i.e., Erithacus and Luscinia). The prevalence of Campylobacter infection in wild birds thus
Engberg, J.; Aarestrup, F. M.; Taylor, D. E.; Gerner-Smidt, P.; Nachamkin, I.
The incidence of human Campylobacter jejuni and C. coli infections has increased markedly in many parts of the world in the last decade as has the number of quinolone-resistant and, to a lesser extent, macrolide-resistant Campylobacter strains causing infections. We review macrolide and quinolone resistance in Campylobacter and track resistance trends in human clinical isolates in relation to use of these agents in food animals. Susceptibility data suggest that erythromycin and other macrolides should remain the drugs of choice in most regions, with systematic surveillance and control measures maintained, but fluoroquinolones may now be of limited use in the empiric treatment of Campylobacter infections in many regions. PMID:11266291
Colles, Frances M; McCarthy, Noel D; Sheppard, Samuel K; Layton, Ruth; Maiden, Martin C J
Relatively little is known about the Campylobacter genotypes colonizing extensively reared broiler flocks and their survival through the slaughter process, despite the increasing demand for free-range and organic products by the consumer. Campylobacter isolates from a free-range boiler flock, sampled before and after slaughter, were genotyped by MLST (multilocus sequence typing) and sequence analysis of the flaA short variable region (SVR). The Campylobacter genotypes isolated before and after slaughter were diverse, with up to five sequence types (STs) (seven-locus allelic profiles resulting from MLST) identified per live bird, up to eight STs identified per carcass and 31 STs identified in all. The majority (72.0%) of isolates sampled from carcasses post-slaughter were indistinguishable from those isolated from the live flock before slaughter by ST and flaA SVR type, however, sampling 'on-farm' failed to capture all of the diversity seen post-slaughter. There were statistically significant increases in the genetic diversity of Campylobacter (p=0.005) and the proportion of C. coli (p=0.002), with some evidence for differential survival of genotypes contaminating the end product. C. coli genotypes isolated after slaughter were more similar to those from free-range and organic meat products sampled nationally, than from the live flock sampled previously. This study demonstrated the utility of MLST in detecting genetic diversity before and after the slaughter process.
Lehtopolku, Mirva; Kotilainen, Pirkko; Puukka, Pauli; Nakari, Ulla-Maija; Siitonen, Anja; Eerola, Erkki; Huovinen, Pentti; Hakanen, Antti J
The agar dilution method has been standardized by the CLSI for the susceptibility testing of Campylobacter species, and according to these standards, the disk diffusion method should be used only in screening for macrolide and ciprofloxacin resistance. Nevertheless, the disk diffusion test is currently widely used, since it is easy to perform in clinical microbiology laboratories. In this study, the disk diffusion method was compared to the agar dilution method by analyzing the in vitro activities of seven antimicrobial agents against 174 Campylobacter strains collected in Finland between 2003 and 2008. Recommendations of the CLSI were followed using Mueller-Hinton agar plates with 5% of sheep blood. For each strain, the disk diffusion tests were performed two to four times. Of the 33 erythromycin-resistant strains (MIC, ≥16 μg/ml), 24 (73%) constantly showed a 6-mm erythromycin inhibition zone (i.e., no inhibition), while for seven strains the inhibition zone varied from 6 to 44 mm in repeated measurements. Among the 141 erythromycin-susceptible strains (MIC, <16 μg/ml), erythromycin inhibition zones varied between 6 and 61 mm. Of the 87 ciprofloxacin-resistant strains, 47 (54%) showed 6-mm inhibition zones, while 40 strains showed inhibition zones between 6 and 60 mm. Significant differences between the repetitions were observed in the disk diffusion for all antimicrobial agents and all strains except for the macrolide-resistant strains regarding the macrolides. For 17 (10%) strains, the variation in repeated measurements was substantial. These results show that the disk diffusion method may not be a reliable tool for the susceptibility testing of Campylobacter spp. Further studies are needed to assess whether the disk diffusion test could be improved or whether all susceptibilities of campylobacters should be tested using an MIC-based method.
Iraola, Gregorio; Pérez, Ruben; Naya, Hugo; Paolicchi, Fernando; Harris, David; Lawley, Trevor D.; Rego, Natalia; Hernández, Martín; Calleros, Lucía; Carretto, Luis; Velilla, Alejandra; Morsella, Claudia; Méndez, Alejandra
Campylobacter fetus subsp. venerealis is the causative agent of bovine genital campylobacteriosis, a sexually transmitted disease distributed worldwide. Campylobacter fetus subsp. venerealis biovar Intermedius strains differ in their biochemical behavior and are prevalent in some countries. We report the first genome sequence for this biovar, isolated from bull prepuce. PMID:23908278
Iraola, Gregorio; Pérez, Ruben; Naya, Hugo; Paolicchi, Fernando; Harris, David; Lawley, Trevor D; Rego, Natalia; Hernández, Martín; Calleros, Lucía; Carretto, Luis; Velilla, Alejandra; Morsella, Claudia; Méndez, Alejandra; Gioffre, Andrea
Campylobacter fetus subsp. venerealis is the causative agent of bovine genital campylobacteriosis, a sexually transmitted disease distributed worldwide. Campylobacter fetus subsp. venerealis biovar Intermedius strains differ in their biochemical behavior and are prevalent in some countries. We report the first genome sequence for this biovar, isolated from bull prepuce.
Introduction: The importance of Salmonella, Campylobacter, E.coli, and Enterococcus as carriers of antimicrobial resistance is well known, but limited work has been done to examine the relationship between this phenotypic characteristic and genotypic attributes among strains isolated in similar set...
Our objective was to identify temperature-related risk factors associated with the colonization of broiler-chicken flocks with Campylobacter spp. in Iceland, with an underlying assumption that at minimum ambient temperatures, flies (Musca domestica) play a role in the epidemiology and seasonality of...
García, Patricia C; Valenzuela, Natalia S; Rodríguez, M Victoria L; León, Eugenia C; Fernández, Heríberto J
Campylobacter jejuni is a common agent of enterocolitis in humans. Campylobacteriosis has been recognized as a zoonotic disease whose reservoir is the intestinal flora of poultry. The reposition of fluid and electrolytes is the recommended treatment, and antimicrobials are required only in severe and/or in prolonged disease. Given the emergence of resistance to drugs commonly used in the treatment of acute diarrhea, we studied the antimicrobial susceptibility of 73 strains of Campylobacter jejuni isolated from stool culture. The antimicrobials tested were: erythromycin, azithromycin, ampicillin and ciprofloxacin. Of the 73 strains tested by E-test, 32.4% were resistant to ciprofloxacin and 6.4% were resistant to ampicillin. Resistance to erythromycin and azithromycin was not detected. The surveillance of antimicrobial resistance of Campylobacter jejuni is important in the evaluation of empirically used antimicrobials in the treatment of bacterial enterocolitis.
Arsi, K; Donoghue, A M; Woo-Ming, A; Blore, P J; Donoghue, D J
Campylobacter is a leading cause of foodborne illness worldwide. It is common in poultry, and human infections are often associated with consumption of contaminated poultry products. One strategy to reduce Campylobacter colonization in poultry is the use of oral probiotics, but this produces variable results, possibly because the probiotics are destroyed in the stomach's acidic environment. Protection (e.g., encapsulation) of isolates may overcome this problem, but there is no assurance that these isolates will have efficacy in the lower gastrointestinal tract. Therefore, screening candidate isolates by directly placing them in the lower intestinal tract via cloacal inoculation may eliminate the time and expense of encapsulating ineffective isolates. Thus, the purpose of this study was to collect bacterial isolates with anti-Campylobacter activity in vitro and evaluate their efficacy in vivo upon either oral or intracloacal administration. Bacterial isolates were collected from healthy birds and were evaluated for efficacy against C. jejuni in vitro. Isolates having generally regarded as safe status and demonstrating in vitro anti-Campylobacter properties were evaluated after oral or intracloacal inoculation into chicks on day 1 (n = 10 birds per isolate per route of administration). On day 7, birds were dosed by oral gavage with a four-strain mixture of wild-type Campylobacter containing at least 1 × 10(7) CFU/ml organisms. On day 14, birds were euthanized and the ceca were collected aseptically for Campylobacter enumeration. When dosed orally, only one isolate had a 1-log reduction in cecal Campylobacter counts, whereas when administered intracloacally, six of these isolates produced a 1- to 3-log reduction in cecal Campylobacter counts in 14-day-old chickens. These results support the strategy of evaluating the efficacy of potential probiotic isolates via cloacal inoculation prior to undergoing the effort of encapsulating isolates for oral administration.
Owens, Jane; Barton, Mary D; Heuzenroeder, Michael W
Six hundred and sixty one samples - primarily fresh chicken faeces - were processed to isolate wild type Campylobacter jejuni bacteriophages, via overlay agar methods using C. jejuni NCTC 12662. The aims of this study were to isolate and purify bacteriophages and then test for their ability to lyse field strains of C. jejuni in vitro. Of all samples processed, 130 were positive for bacteriophages. A distinct difference was observed between samples from different poultry enterprises. No bacteriophages could be isolated from indoor broilers. The majority of bacteriophages were isolated from free range poultry - both broilers and egg layers. Bacteriophages were purified and then selected for characterization based on their ability to produce clear lysis on plaque assay, as opposed to turbid plaques. Two hundred and forty one C. jejuni field isolates were tested for sensitivity to the bacteriophages. Lysis was graded subjectively and any minimal lysis was excluded. Using this system, 59.0% of the C. jejuni isolates showed significant sensitivity to at least one bacteriophage. The sensitivity to individual bacteriophages ranged from 10.0% to 32.5% of the C. jejuni isolates. Five bacteriophages were examined by electron microscopy and determined to belong to the Myoviridae family. The physical size, predicted genetic composition and genome size of the bacteriophages correlated well with other reported Campylobacter bacteriophages. The reasons for the observed difference between indoor broilers and free range poultry is unknown, but are postulated to be due to differences in the Campylobacter population in birds under different rearing conditions.
Dipineto, Ludovico; Borrelli, Luca; Pace, Antonino; Romano, Violante; D'Orazio, Stefano; Varriale, Lorena; Russo, Tamara Pasqualina; Fioretti, Alessandro
Avian species are considered as the main reservoir of Campylobacter spp. However, few data are available on the presence of this microorganism in pet birds. This study was therefore performed to determine the prevalence of Campylobacter spp. in pet birds bred in southern Italy. Faecal samples were collected from 88 cages housing different species of pet birds and examined by bacteriological culture and polymerase chain reaction. A total of 13.6% of the cage samples were positive for Campylobacter coli. Other Campylobacter spp. were not found. The study shows that C. coli can be isolated from the cages of apparently healthy pet birds, which should therefore be considered as potential carriers of C. coli and a possible source of infection for humans and companion animals.
Hannon, Sherry J; Allan, Brenda; Waldner, Cheryl; Russell, Margaret L; Potter, Andrew; Babiuk, Lorne A; Townsend, Hugh G G
This fecal prevalence study targeted cattle from 7 large (10,000 to > 40,000 head) commercial feedlots in Alberta as a means of establishing Campylobacter levels in cattle just prior to animals entering the food chain. Overall, 87% [95% confidence interval (CI) = 86-88] of 2776 fresh pen-floor fecal samples were culture positive for Campylobacter species, with prevalences ranging from 76% to 95% among the 7 feedlots. Campylobacter spp. prevalence was 88% (95% CI = 86-90) in the summer (n = 1376) and 86% (95% CI = 85-88) in the winter (n = 1400). In addition, 69% (95% CI = 66-71) of 1486 Campylobacter spp. positive samples were identified as Campylobacter jejuni using hippurate hydrolysis testing. Of those, 64% (95% CI = 58-70) of 277 and 70% (95% CI = 67-72) of 1209 Campylobacter isolates were identified as C. jejuni in winter and summer, respectively. After accounting for clustering within pen and feedlot, feedlot size and the number of days on feed were associated with Campylobacter spp. isolation rates. The high isolation rates of Campylobacter spp. and C. jejuni in feedlot cattle feces in this study suggest a potential role for feedlot cattle in the complex epidemiology of campylobacters in Alberta.
Uyttendaele, M; Baert, K; Ghafir, Y; Daube, G; De Zutter, L; Herman, L; Dierick, K; Pierard, D; Dubois, J J; Horion, B; Debevere, J
The objective of this study was to do an exercise in risk assessment on Campylobacter spp. for poultry based meat preparations in Belgium. This risk assessment was undertaken on the demand of the competent national authorities as one of the supportive factors to define risk-based microbiological criteria. The quantitative risk assessment model follows a retail to table approach and is divided in different modules. The contamination of raw chicken meat products (CMPs) was represented by a normal distribution of the natural logarithm of the concentration of Campylobacter spp. (ln[Camp]) in raw CMPs based on data from surveillance programs in Belgium. To analyse the relative impact of reducing the risk of campylobacteriosis associated with a decrease in the Campylobacter contamination level in these types of food products, the model was run for different means and standard deviations of the normal distribution of the ln[Camp] in raw CMPs. The limitation in data for the local situation in Belgium and on this particular product and more precisely the semi-quantitative nature of concentration of Campylobacter spp. due to presence/absence testing, was identified as an important information gap. Also the knowledge on the dose-response relationship of Campylobacter spp. was limited, and therefore three different approaches of dose-response modelling were compared. Two approaches (1 and 2), derived from the same study, showed that the reduction of the mean of the distribution representing the ln[Camp] in raw CMPs is the best approach to reduce the risk of Campylobacter spp. in CMPs. However, for the simulated exposure and approach 3 it was observed that the reduction of the standard deviation is the most appropriate technique to lower the risk of campylobacteriosis. Since the dose-response models used in approach 1 and 2 are based on limited data and the reduction of the mean corresponds with a complete shift of the contamination level of raw CMPs, demanding high efforts
Doern, C.; Fader, R.; Ferraro, M. J.; Pillai, D. R.; Rychert, J.; Doyle, L.; Lainesse, A.; Karchmer, T.; Mortensen, J. E.
Diarrhea due to enteric bacterial pathogens causes significant morbidity and mortality in the United States and worldwide. However, bacterial pathogens may be infrequently identified. Currently, culture and enzyme immunoassays (EIAs) are the primary methods used by clinical laboratories to detect enteric bacterial pathogens. We conducted a multicenter evaluation of the BD Max enteric bacterial panel (EBP) PCR assay in comparison to culture for the detection of Salmonella spp., Shigella spp., Campylobacter jejuni, and Campylobacter coli and an EIA for Shiga toxins 1 and 2. A total of 4,242 preserved or unpreserved stool specimens, including 3,457 specimens collected prospectively and 785 frozen, retrospective samples, were evaluated. Compared to culture or EIA, the positive percent agreement (PPA) and negative percent agreement (NPA) values for the BD Max EBP assay for all specimens combined were as follows: 97.1% and 99.2% for Salmonella spp., 99.1% and 99.7% for Shigella spp., 97.2% and 98.4% for C. jejuni and C. coli, and 97.4% and 99.3% for Shiga toxins, respectively. Discrepant results for prospective samples were resolved with alternate PCR assays and bidirectional sequencing of amplicons. Following discrepant analysis, PPA and NPA values were as follows: 97.3% and 99.8% for Salmonella spp., 99.2% and 100% for Shigella spp., 97.5% and 99.0% for C. jejuni and C. coli, and 100% and 99.7% for Shiga toxins, respectively. No differences in detection were observed for samples preserved in Cary-Blair medium and unpreserved samples. In this large, multicenter study, the BD Max EBP assay showed superior sensitivity compared to conventional methods and excellent specificity for the detection of enteric bacterial pathogens in stool specimens. PMID:25740779
Multicenter evaluation of the BD max enteric bacterial panel PCR assay for rapid detection of Salmonella spp., Shigella spp., Campylobacter spp. (C. jejuni and C. coli), and Shiga toxin 1 and 2 genes.
Harrington, S M; Buchan, B W; Doern, C; Fader, R; Ferraro, M J; Pillai, D R; Rychert, J; Doyle, L; Lainesse, A; Karchmer, T; Mortensen, J E
Diarrhea due to enteric bacterial pathogens causes significant morbidity and mortality in the United States and worldwide. However, bacterial pathogens may be infrequently identified. Currently, culture and enzyme immunoassays (EIAs) are the primary methods used by clinical laboratories to detect enteric bacterial pathogens. We conducted a multicenter evaluation of the BD Max enteric bacterial panel (EBP) PCR assay in comparison to culture for the detection of Salmonella spp., Shigella spp., Campylobacter jejuni, and Campylobacter coli and an EIA for Shiga toxins 1 and 2. A total of 4,242 preserved or unpreserved stool specimens, including 3,457 specimens collected prospectively and 785 frozen, retrospective samples, were evaluated. Compared to culture or EIA, the positive percent agreement (PPA) and negative percent agreement (NPA) values for the BD Max EBP assay for all specimens combined were as follows: 97.1% and 99.2% for Salmonella spp., 99.1% and 99.7% for Shigella spp., 97.2% and 98.4% for C. jejuni and C. coli, and 97.4% and 99.3% for Shiga toxins, respectively. Discrepant results for prospective samples were resolved with alternate PCR assays and bidirectional sequencing of amplicons. Following discrepant analysis, PPA and NPA values were as follows: 97.3% and 99.8% for Salmonella spp., 99.2% and 100% for Shigella spp., 97.5% and 99.0% for C. jejuni and C. coli, and 100% and 99.7% for Shiga toxins, respectively. No differences in detection were observed for samples preserved in Cary-Blair medium and unpreserved samples. In this large, multicenter study, the BD Max EBP assay showed superior sensitivity compared to conventional methods and excellent specificity for the detection of enteric bacterial pathogens in stool specimens.
Ahmed, Heba A; El Hofy, Fatma I; Ammar, Ahmed M; Abd El Tawab, Ashraf A; Hefny, Ahmed A
The public health importance of the genus Campylobacter is attributed to several species causing diarrhea in consumers. Poultry and their meat are considered the most important sources of human campylobacteriosis. In this study, 287 samples from chicken (131 cloacal swabs, 39 chicken skin, 78 chicken meat, and 39 cecal parts) obtained from retail outlets as well as 246 stool swabs from gastroenteritis patients were examined. A representative number of the biochemically identified Campylobacter jejuni isolates were identified by real-time PCR, confirming the identification of the isolates as C. jejuni. Genotyping of the examined isolates (n = 31) by enterobacterial repetitive intergenic consensus PCR (ERIC-PCR) revealed a high discriminatory index of ERIC-PCR (D = 0.948), dividing C. jejuni isolates of chicken and human origins into 18 profiles and four clusters. The 18 profiles obtained indicated the heterogeneity of C. jejuni. Dendrogram analysis showed that four clusters were generated; all human isolates fell into clusters I and III. These observations further support the existence of a genetic relationship between human and poultry isolates examined in the present study. In conclusion, the results obtained support the speculation that poultry and poultry meat have an important role as sources of infection in the acquisition of Campylobacter infection in humans.
Hoelzl, C; Mayerhofer, U; Steininger, M; Brüller, W; Hofstädter, D; Aldrian, U
Campylobacter infections are one of the most prominent worldwide food-related diseases. The primary cause of these infections is reported to be improper food handling, in particular cross-contamination during domestic preparation of raw chicken products. In the present study, food handling behaviors in Austria were surveyed and monitored, with special emphasis on Campylobacter cross-contamination. Forty participants (25 mothers or fathers with at least one child ≤10 years of age and 15 elderly persons ≥60 years of age) were observed during the preparation of a chicken salad (chicken slices plus lettuce, tomato, and cucumber) using a direct structured observational scoring system. The raw chicken carcasses and the vegetable part of the salad were analyzed for Campylobacter. A questionnaire concerning knowledge, attitudes, and interests related to food safety issues was filled out by the participants. Only 57% of formerly identified important hygiene measures were used by the participants. Deficits were found in effective hand washing after contact with raw chicken meat, but proper changing and cleaning of the cutting board was noted. Campylobacter was present in 80% of raw chicken carcasses, albeit the contamination rate was generally lower than the limit of quantification (10 CFU/g). In the vegetable part of the prepared product, no Campylobacter was found. This finding could be due to the rather low Campylobacter contamination rate in the raw materials and the participants' use of some important food handling behaviors to prevent cross-contamination. However, if the initial contamination had been higher, the monitored deficits in safe food handling could lead to quantifiable risks, as indicated in other published studies. The results of the observational trial and the questionnaire indicated knowledge gaps in the food safety sector, suggesting that further education of the population is needed to prevent the onset of foodborne diseases.
Frey, Steven K; Topp, Edward; Khan, Izhar U H; Ball, Bonnie R; Edwards, Mark; Gottschall, Natalie; Sunohara, Mark; Lapen, David R
This work investigated chlortetracycline, tylosin, and tetracycline (plus transformation products), and DNA-based quantitative Campylobacter spp. and Campylobacter tetracycline antibiotic resistant genes (tet(O)) in tile drainage, groundwater, and soil before and following a liquid swine manure (LSM) application on clay loam plots under controlled (CD) and free (FD) tile drainage. Chlortetracycline/tetracycline was strongly bound to manure solids while tylosin dominated in the liquid portion of manure. The chlortetracycline transformation product isochlortetracycline was the most persistent analyte in water. Rhodamine WT (RWT) tracer was mixed with manure and monitored in tile and groundwater. RWT and veterinary antibiotic (VA) concentrations were strongly correlated in water which supported the use of RWT as a surrogate tracer. While CD reduced tile discharge and eliminated application-induced VA movement (via tile) to surface water, total VA mass loading to surface water was not affected by CD. At both CD and FD test plots, the biggest 'flush' of VA mass and highest VA concentrations occurred in response to precipitation received 2d after application, which strongly influenced the flow abatement capacity of CD on account of highly elevated water levels in field initiating overflow drainage for CD systems (when water level <0.3m below surface). VA concentrations in tile and groundwater became very low within 10d following application. Both Campylobacter spp. and Campylobacter tet(O) genes were present in groundwater and soil prior to application, and increased thereafter. Unlike the VA compounds, Campylobacter spp. and Campylobacter tet(O) gene loadings in tile drainage were reduced by CD, in relation to FD.
Sáenz, Yolanda; Zarazaga, Myriam; Lantero, Marta; Gastañares, M. José; Baquero, Fernando; Torres, Carmen
Colonization by Campylobacter strains was investigated in human, broiler, and pig fecal samples from 1997- 1998, as well as in foods of animal origin, and antibiotic susceptibility testing was carried out for these strains. Campylobacter strains were isolated in the foods of animal origin (55 of 101 samples; 54.4%), intestinal samples from broilers (85 of 105; 81%), and pigs (40 of 45; 88.9%). A total of 641 Campylobacter strains were isolated from 8,636 human fecal samples of clinical origin (7.4%). Campylobacter jejuni was the most frequently isolated species from broilers (81%) and humans (84%), and Campylobacter coli was most frequently isolated from pigs (100%). An extremely high frequency of ciprofloxacin resistance was detected among Campylobacter strains, particularly those isolated from broilers and pigs (99%), with a slightly lower result for humans (72%); cross-resistance with nalidixic acid was almost always observed. A higher frequency of resistance to erythromycin (81.1%), ampicillin (65.7%), gentamicin (22.2%), and amikacin (21.6%) was detected in C. coli strains isolated from pigs compared to those isolated from humans (34.5, 29.3, 8.6, and 0%, respectively). A low frequency of erythromycin resistance was found in C. jejuni or C. coli isolated from broilers. A greater resistance to ampicillin and gentamicin (47.4 and 11.9%, respectively) was detected in C. jejuni isolated from broilers than in human strains (38 and 0.4%, respectively). β-Lactamase production was found in 81% of the Campylobacter strains tested, although 44% of them were characterized as ampicillin susceptible. The increasing rates of Campylobacter resistance make advisable a more conservative policy for the use of antibiotics in farm animals. PMID:10639348
Oh, Jae-Young; Kwon, Yong-Kuk; Wei, Bai; Jang, Hyung-Kwan; Lim, Suk-Kyung; Kim, Cheon-Hyeon; Jung, Suk-Chan; Kang, Min-Su
Thirty-nine human isolates of Campylobacter jejuni obtained from a national university hospital during 2007-2010 and 38 chicken isolates of C. jejuni were collected from poultry farms during 2009-2010 in South Korea were used in this study. Campylobacter genomic species and virulence-associated genes were identified by PCR. Pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST) were performed to compare their genetic relationships. All isolates were highly resistant to ciprofloxacin, nalidixic acid, and tetracycline. Of all isolates tested, over 94% contained seven virulence associated genes (flaA, cadF, racR, dnaJ, cdtA, cdtB, and cdtC). All isolates were classified into 39 types by PFGE clustering with 90% similarity. Some chicken isolates were incorporated into some PFGE types of human isolates. MLST analysis for the 39 human isolates and 38 chicken isolates resulted in 14 and 23 sequence types (STs), respectively, of which 10 STs were new. STs overlapped in both chicken and human isolates included ST-21, ST-48, ST-50, ST-51, and ST-354, of which ST-21 was the predominant ST in both human and chicken isolates. Through combined analysis of PFGE types and STs, three chicken isolates were clonally related to the three human isolates associated with food poisoning (VII-ST-48, XXII-ST-354, and XXVIII-ST-51). They were derived from geographically same or distinct districts. Remarkably, clonal spread of food poisoning pathogens between animals and humans was confirmed by population genetic analysis. Consequently, contamination of campylobacters with quinolone resistance and potential virulence genes in poultry production and consumption may increase the risk of infections in humans.
Duffy, Lesley L; Blackall, Patrick J; Cobbold, Rowland N; Fegan, Narelle
Poultry are considered a major source for campylobacteriosis in humans. A total of 1866 Campylobacter spp. isolates collected through the poultry processing chain were typed using flaA-restriction fragment length polymorphism to measure the impact of processing on the genotypes present. Temporally related human clinical isolates (n = 497) were also typed. Isolates were obtained from whole chicken carcass rinses of chickens collected before scalding, after scalding, before immersion chilling, after immersion chilling and after packaging as well as from individual caecal samples. A total of 32 genotypes comprising at least four isolates each were recognised. Simpson's Index of Diversity (D) was calculated for each sampling site within each flock, for each flock as a whole and for the clinical isolates. From caecal collection to after packaging samples the D value did not change in two flocks, decreased in one flock and increased in the fourth flock. Dominant genotypes occurred in each flock but their constitutive percentages changed through processing. There were 23 overlapping genotypes between clinical and chicken isolates. The diversity of Campylobacter is flock dependant and may alter through processing. This study confirms that poultry are a source of campylobacteriosis in the Australian population although other sources may contribute.
Kovačić, Ana; Carev, Merica; Tripković, Ingrid; Srečec, Siniša; Siško-Kraljević, Katarina
Consumption of poultry is considered to be an important source of human infection with Campylobacter. In the period from 2008 to 2010, 50 isolates of Campylobacter jejuni from human faeces were analysed and compared with 61 isolates from poultry by pulsed field gel electrophoresis using SmaI and KpnI. Based on the analysis of SmaI macrorestriction profiles, 86 isolates (77.5 %) were assigned to 15 S clusters: 31 (62 %) from humans and 55 from poultry (90.2 %). Altogether 21 isolates (19 %) exhibited macrorestriction profiles common to both humans and poultry after restriction with SmaI and KpnI. A total of five identical pulsotypes were isolated from both poultry and patients and one of them appeared in eight different locations in the time interval of one year. These results indicate that poultry could be an important source of Campylobacter infection in Split and Dalmatia County which is the biggest County in Croatia and the most important tourist destination.
Simultaneous occurrence of Salmonella enterica, Campylobacter spp. and Yersinia enterocolitica along the pork production chain from farm to meat processing in five conventional fattening pig herds in Lower Saxony.
Niemann, Jana-Kristin; Alter, Thomas; Gölz, Greta; Tietze, Erhard; Fruth, Angelika; Rabsch, Wolfgang; von Münchhausen, Christiane; Merle, Roswitha; Kreienbrock, Lothar
The objectives of this study were to gather data on the occurrence of Salmonella (S.) enterica, Campylobacter spp. and Yersinia (Y.) enterocolitica along the pork production chain and to further analyze detected Salmonella isolates by additionally applying MLVA (multiple-locus variable-number tandem repeat analysis). In total, 336 samples were collected at primary production, slaughter and meat processing from five conventional fattening pig farms and one common slaughterhouse. At farm level, S. enterica, Campylobacter spp. and Y. enterocolitica were detected in 19.4%, 38.9% and 11.1% of pooled fecal samples of fattening pigs. At slaughter, more than two-thirds of examined carcasses, 24% of carcass surfaces samples and about 60% of cecal content samples were positive for at least one of the examined pathogens. An amount of 4% of meat samples were positive for non-human pathogenic Y. enterocolitica. Identical MLVA patterns of Salmonella isolates from farm- and associated slaughterhouse samples demonstrated transmission across both production stages. Other MLVA patterns found at slaughter indicated possible colonization of pigs during transport or lairage and/or cross-contamination during slaughter. Identical MLVA patterns from risk tissues and the nearby carcass surface evidenced a direct contamination of carcasses as well. Overall, our data showed wide distribution ranges for all three examined pathogens within the pig production chain and underline the need for appropriate intervention strategies at pre- and postharvest.
Using rDNA sequencing analysis, we examined the bacterial diversity and the presence of opportunistic bacterial pathogens (i.e., Campylobacter and Helicobacter) in Red Knot (Calidris canutus, n=40), Ruddy Turnstone (Arenaria interpres, n=35), and Semipalmated Sandpiper (Calidris ...
Saludes, Paula; Araguás, Cristina; Sánchez-Delgado, Jordi; Dalmau, Blai; Font, Bernat
The isolation of Candida spp. in ascites of cirrhotic patients is an uncommon situation in clinical practice. Factors that have been associated with increased susceptibility to primary fungal peritonitis are exposure to broad-spectrum antibiotics and immunosuppression, a typical situation of these patients. We report seven episodes of Candida spp. isolation in ascites of cirrhotic patients detected in our hospital during the past 15years.
Gonsalves, Camila Cristina; Borsoi, Anderlise; Perdoncini, Gustavo; Rodrigues, Laura Beatriz; do Nascimento, Vladimir Pinheiro
Campylobacter spp. cause foodborne illnesses in humans primarily through the consumption of contaminated chicken. The aim of this study was to evaluate the United States Department of Agriculture's (USDA) recommended methodology, protocol MLG 41.02, for the isolation, identification and direct plate counting of Campylobacter jejuni and C. coli samples from the broiler slaughtering process. A plating method using both mCCDA and Campy-Cefex agars is recommended to recover Campylobacter cells. It is also possible to use this method in different matrices (cloacal swabs and water samples). Cloacal swabs, samples from pre-chiller and post-chiller carcasses and samples of pre-chiller, chiller and direct supply water were collected each week for four weeks from the same flock at a slaughterhouse located in an abattoir in southern Brazil. Samples were analyzed to directly count Campylobacter spp., and the results showed a high frequency of Campylobacter spp. on Campy-Cefex agar. For the isolated species, 72% were identified as Campylobacter jejuni and 38% as Campylobacter coli. It was possible to count Campylobacter jejuni and Campylobacter coli from different samples, including the water supply samples, using the two-agar method. These results suggest that slaughterhouses can use direct counting methods with both agars and different matrices as a monitoring tool to assess the presence of Campylobacter bacteria in their products.
Di Giannatale, Elisabetta; Di Serafino, Gabriella; Zilli, Katiuscia; Alessiani, Alessandra; Sacchini, Lorena; Garofolo, Giuliano; Aprea, Giuseppe; Marotta, Francesca
Campylobacter has developed resistance to several antimicrobial agents over the years, including macrolides, quinolones and fluoroquinolones, becoming a significant public health hazard. A total of 145 strains derived from raw milk, chicken faeces, chicken carcasses, cattle faeces and human faeces collected from various Italian regions, were screened for antimicrobial susceptibility, molecular characterization (SmaI pulsed-field gel electrophoresis) and detection of virulence genes (sequencing and DNA microarray analysis). The prevalence of C. jejuni and C. coli was 62.75% and 37.24% respectively. Antimicrobial susceptibility revealed a high level of resistance for ciprofloxacin (62.76%), tetracycline (55.86%) and nalidixic acid (55.17%). Genotyping of Campylobacter isolates using PFGE revealed a total of 86 unique SmaI patterns. Virulence gene profiles were determined using a new microbial diagnostic microarray composed of 70-mer oligonucleotide probes targeting genes implicated in Campylobacter pathogenicity. Correspondence between PFGE and microarray clusters was observed. Comparisons of PFGE and virulence profiles reflected the high genetic diversity of the strains examined, leading us to speculate different degrees of pathogenicity inside Campylobacter populations. PMID:24556669
Di Giannatale, Elisabetta; Di Serafino, Gabriella; Zilli, Katiuscia; Alessiani, Alessandra; Sacchini, Lorena; Garofolo, Giuliano; Aprea, Giuseppe; Marotta, Francesca
Campylobacter has developed resistance to several antimicrobial agents over the years, including macrolides, quinolones and fluoroquinolones, becoming a significant public health hazard. A total of 145 strains derived from raw milk, chicken faeces, chicken carcasses, cattle faeces and human faeces collected from various Italian regions, were screened for antimicrobial susceptibility, molecular characterization (SmaI pulsed-field gel electrophoresis) and detection of virulence genes (sequencing and DNA microarray analysis). The prevalence of C. jejuni and C. coli was 62.75% and 37.24% respectively. Antimicrobial susceptibility revealed a high level of resistance for ciprofloxacin (62.76%), tetracycline (55.86%) and nalidixic acid (55.17%). Genotyping of Campylobacter isolates using PFGE revealed a total of 86 unique SmaI patterns. Virulence gene profiles were determined using a new microbial diagnostic microarray composed of 70-mer oligonucleotide probes targeting genes implicated in Campylobacter pathogenicity. Correspondence between PFGE and microarray clusters was observed. Comparisons of PFGE and virulence profiles reflected the high genetic diversity of the strains examined, leading us to speculate different degrees of pathogenicity inside Campylobacter populations.
Campylobacter jejuni, a Gram-negative rod, is a zoonotic pathogen associated with human acute bacterial gastroenteritis worldwide. Poultry products are regarded as a major source of this bacterium for human infection. Although this bacterium is a commensal in chicken cecal microbiome, Campylobacte...
Castillo-Martínez, M L; Sánchez-Sánchez, S; Rodríguez-Montaño, R; Quiñones-Ramírez, E I; Lugo de la Fuente, G; Vázquez-Salinas, C
The human gastroenteritis caused by Campylobacter jejuni in some industrialized countries is higher than gastroenteritis produced by Salmonella and Shigella. This has induced the development of techniques to demonstrate the presence of the microorganism in different foods using some culture media combinations. There is not a method to isolate C. jejuni from roasted chicken and fried pork meat, which are popular foods in México. The sensitivity of two culture media combinations was compared: Rama broth (RB)-Rama agar (RA) and Preston broth (PB)-Skirrow agar (SA) to isolate C. jejuni from these foods. The RB-RA combination demonstrated to be the best one to isolate C. jejuni.
Waldenström, Jonas; Mevius, Dik; Veldman, Kees; Broman, Tina; Hasselquist, Dennis; Olsen, Björn
In order to determine the occurrence and frequency of resistant strains of the bacterium Campylobacter jejuni and to establish baseline MICs in isolates from an environmental reservoir, the resistance profiles of 10 antimicrobial substances were determined for 137 C. jejuni isolates from wild birds in Sweden. Observed MICs were generally low, with only low to moderate incidence of resistance to the tested compounds. One isolate, however, was resistant to nalidixic acid and ciprofloxacin, indicating that quinolone-resistant genotypes of C. jejuni have the potential to spread to wild bird hosts. PMID:15870331
Newell, D. G.; Elvers, K. T.; Dopfer, D.; Hansson, I.; Jones, P.; James, S.; Gittins, J.; Stern, N. J.; Davies, R.; Connerton, I.; Pearson, D.; Salvat, G.; Allen, V. M.
The prevention and control of Campylobacter colonization of poultry flocks are important public health strategies for the control of human campylobacteriosis. A critical review of the literature on interventions to control Campylobacter in poultry on farms was undertaken using a systematic approach. Although the focus of the review was on aspects appropriate to the United Kingdom poultry industry, the research reviewed was gathered from worldwide literature. Multiple electronic databases were employed to search the literature, in any language, from 1980 to September 2008. A primary set of 4,316 references was identified and scanned, using specific agreed-upon criteria, to select relevant references related to biosecurity-based interventions. The final library comprised 173 references. Identification of the sources of Campylobacter in poultry flocks was required to inform the development of targeted interventions to disrupt transmission routes. The approach used generally involved risk factor-based surveys related to culture-positive or -negative flocks, usually combined with a structured questionnaire. In addition, some studies, either in combination or independently, undertook intervention trials. Many of these studies were compromised by poor design, sampling, and statistical analysis. The evidence for each potential source and route of transmission on the poultry farm was reviewed critically, and the options for intervention were considered. The review concluded that, in most instances, biosecurity on conventional broiler farms can be enhanced and this should contribute to the reduction of flock colonization. However, complementary, non-biosecurity-based approaches will also be required in the future to maximize the reduction of Campylobacter-positive flocks at the farm level. PMID:21984249
Mokracka, Joanna; Koczura, Ryszard; Kaznowski, Adam
We analyzed the ability of extraintestinal strains of Enterobacter spp. and Citrobacter spp. to employ different siderophore-mediated strategies of iron acquisition. All strains produced iron-chelating compounds. Cross-feeding assays indicated that most isolates of both Enterobacter spp. and Citrobacter spp. excreted catecholate siderophore enterobactin, less produced aerobactin, and single strains excreted hydroxamates different from aerobactin. Besides, we analyzed if the strains had the ability to produce the siderophore yersiniabactin coded by the Yersinia high-pathogenicity island (HPI). The presence of HPI genes was observed in single isolates of three species: E. cloaceae, E. aerogenes and C. koseri. A detailed polymerase chain reaction analysis revealed differences in the genetic organization of the HPIs; however, in a cross-feeding test we proved that yersiniabactin was produced and the island was functional.
Hansson, I; Nyman, A; Lahti, E; Gustafsson, P; Olsson Engvall, E
A study was performed with the aim to investigate associations between Campylobacter in chicken caecum, carcass skin, underlying breast muscle and packaged breast fillets. Samples were taken from 285 chickens from 57 flocks and analysed according to ISO 10272. Campylobacter spp. were isolated from caecal samples from 41 flocks. From birds of the same 41 flocks Campylobacter could be detected and quantified in 194 (68%) skin samples. Moreover, Campylobacter spp. were enumerated in 13 (5%) underlying muscle samples originating from 9 of the 41 flocks. The mean number of Campylobacter spp. in the 194 skin samples which could be counted was 2.3 log cfu/g and for the 13 underlying muscle samples 1.3 log cfu/g. Campylobacter could only be quantified in those breast muscle samples with a finding in corresponding skin sample. Five packaged chicken fillets were taken from each 25 of the 57 flocks and analysed both quantitatively and qualitatively. In qualitative analysis Campylobacter was detected in 79 (63%) fillets from 16 flocks and quantified in 24 (19%) samples from 11 flocks. The results showed a significant association (P < 0.05) between findings of Campylobacter on carcass skin (log cfu/g) and the proportion of Campylobacter positive breast muscle samples.
LIM, Suk-Kyung; MOON, Dong-Chan; CHAE, Myung Hwa; KIM, Hae Ji; NAM, Hyang-Mi; KIM, Su-Ran; JANG, Gum-Chan; LEE, Kichan; JUNG, Suk-Chan; LEE, Hee-Soo
Resistance to antimicrobials was measured in 73 isolates of Campylobacter jejuni (C. jejuni) and 121 isolates of Campylobacter coli (C. coli) from chicken and swine feces and carcasses in Korea. Both bacterial species showed the highest resistance to (fluoro) quinolones (ciprofloxacin and nalidixic acid) out of the nine antimicrobials tested. Erythromycin resistance was much higher in C. coli (19.0%, 23/121) than in C. jejuni (6.8%, 5/73). The mutation in the 23S rRNA gene was primarily responsible for macrolide resistance in Campylobacter isolates. Several amino acid substitutions in the L4 and L22 ribosomal proteins may play a role in the mechanism of resistance, but the role requires further evaluation. A total of eight virulence genes were detected in 28 erythromycin-resistant Campylobacter isolates. All C. jejuni isolates carried more than four such genes, while C. coli isolates carried fewer than three such genes. The high rate of resistance highlights the need to employ more prudent use of critically important antimicrobials, such as fluoroquinolones and macrolides, in swine and poultry production, and to more carefully monitor antimicrobial resistance in Campylobacter isolates in food animals. PMID:27593510
Rippa, Paola; Battaglia, Luciana; Parisi, Nicola
Cronobacter spp. (Enterobacter sakazakii) is an opportunistic bacterial pathogen capable of causing disease and even fatalities in newborn infants within the first weeks of life if consumed as part of the diet. Premature and immunocompromised newborn infants are at particular risk. The microorganism has been isolated from a variety of foods including contaminated infant milk formula powder and milk powder substitute. The study aimed to evaluate the level of microbiological contamination in 47 samples of mozzarella cheese made with cow’s milk collected from artisan cheese producers in Southern Italy. Samples were collected from commercial sales points and underwent qualitative and quantitative microbiological analyses to test for the bacterial contaminants most commonly found in milk and cheese products. The 47 samples underwent qualitative and quantitative microbiological tests according to ISO UNI EN standards. Analyses focused on Staphylococcus aures, Salmonella spp., Listeria monocytogenes, Pseudomonas spp., E. coli, Yersinia spp., total coliforms and Cronobacter sakazakii. The ISO/TS 22964:2006 method was used to investigate possible contamination by C. sakazakii. Biochemical identification was carried out using an automated system for identification and susceptibility tests. None of the samples examined resulted positive for Salmonella spp. or Listeria spp. Only one sample resulted positive for Staphylococcus aureus. Pseudomonas spp. was isolated in 10 (21%) of 47 samples. High levels of total coliforms were found in 10 of 47 samples. Cronobacter spp. (Enterobacter sakazakii) was isolated in one sample. This is the first study to confirm isolation of C. sakazakii in artisan mozzarella cheese made from cow’s milk. The presence of C. sakazakii could be related to external contamination during the phases of production or to the use of contaminated milk. Since mozzarella is recommended in the diet of children and adults of all ages, this present study helps
Feizabadi, Mohammad Mehdi; Dolatabadi, Samaneh; Zali, Mohammad Reza
To detect campylobacteriosis and determine the drug susceptibility of causative organisms, we acquired 500 diarrheic samples in Cary-Blair transfer medium from two pediatric hospitals in Tehran between October 2004 and October 2005. The samples were also enriched in Preston broth (with supplements) and defibrinated sheep blood (7%). They were plated from both media on Brucella agar containing antibiotics and blood. Isolates were identified through biochemical tests and by the polymerase chain reaction method. Drug susceptibility testing was performed using the disk diffusion method. In total, 40 Campylobacter strains were isolated (8%). C. jejuni was the dominant species (85.8%) followed by C. coli (14.2%). The rates of resistance to antimicrobial agents were as follows: ciprofloxacin (61.7%), ceftazidime (47%), carbenicillin (35%), tetracycline (20.5%), cefotaxime (14.7%), ampicillin (11.7%), neomycin erythromycin and chloramphenicol (2.9%), gentamicin, streptomycin, imipenem and colistin (0.0%). Campylobacter is an important cause of diarrhea among Iranian children. The detection of Campylobacter increases by 25% if samples are treated in enrichment broth prior to plating. The high rate of resistance to ciprofloxacin is alarming, and further investigation into the possible reasons for this is imperative.
Garin, Benoit; Gouali, Malika; Wouafo, Marguerite; Perchec, Anne-Marie; Pham, Minh Thu; Ravaonindrina, Noro; Urbès, Florence; Gay, Manu; Diawara, Abdoulaye; Leclercq, Alexandre; Rocourt, Jocelyne; Pouillot, Régis
Quantitative data on Campylobacter contamination of food are lacking, notably in developing countries. We assessed Campylobacter contamination of chicken neck-skins at points of slaughter in 5 major cities in Africa (Dakar in Senegal, Yaounde in Cameroon), Oceania (Noumea in New Caledonia), the Indian Ocean (Antananarivo in Madagascar) and Asia (Ho Chi Minh City (HCMC) in Vietnam. One hundred and fifty slaughtered chickens were collected in each of the 5 major cities from semi-industrial abattoirs or markets (direct slaughter by the seller), and 65.5% (491/750) were found to be Campylobacter-positive. Two cities, Yaounde and Noumea, demonstrated high prevalence Campylobacter detection rates (92.7% and 96.7% respectively) in contrast with HCMC (15.3%). Four species were identified among 633 isolates, namely C. jejuni (48.3%), C. coli (37.3%), C. lari (11.7%) and C. upsaliensis (1%). HCMC was the only city with C. lari isolation as was Antananarivo for C. upsaliensis. C. coli was highly prevalent only in Yaounde (69.5%). Among the 491 samples positive in Campylobacter detection, 329 were also positive with the enumeration method. The number of Campylobacter colony-forming units (CFU) per gram of neck-skin in samples positive in enumeration was high (mean of the log(10): 3.2 log(10) CFU/g, arithmetic mean: 7900CFU/g). All the cities showed close enumeration means except HCMC with a 1.81 log(10) CFU/g mean for positive samples. Semi-industrial abattoir was linked to a significant lower count of Campylobacter contamination than direct slaughter by the seller (p=0.006). On 546 isolates (546/633, 86.3%) tested for antibiotic susceptibility, resistance to erythromycin, ampicillin and ciprofloxacin was observed for respectively 11%, 19% and 50%. HCMC was the city where antibiotic resistant rates were the highest (95%, p=0.014). Considering the 329 positive chickens in Campylobacter enumeration, the mean number of resistant isolates to at least 2 different antibiotic
Jay-Russell, M T; Bates, A; Harden, L; Miller, W G; Mandrell, R E
We report the isolation of Campylobacter species from the same population of feral swine that was investigated in San Benito County, California, during the 2006 spinach-related Escherichia coli O157:H7 outbreak. This is the first survey of Campylobacter in a free-ranging feral swine population in the United States. Campylobacter species were cultured from buccal and rectal-anal swabs, colonic faeces and tonsils using a combination of selective enrichment and antibiotic-free membrane filtration methods. Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF-MS, Bruker Daltonics, Inc., Billerica, MA, USA) was used to identify species followed by confirmatory multiplex PCR or 16S rRNA sequencing. Genetic relatedness of Campylobacter jejuni and Campylobacter coli strains was determined by multilocus sequence typing (MLST) and porA allele sequencing. Altogether, 12 (40%) of 30 feral swine gastrointestinal and oral cavity specimens were positive, and six species were isolated: Campylobacter coli, Campylobacter fetus, Campylobacter hyointestinalsis, Campylobacter jejuni, Campylobacter lanienae and Campylobacter sputorum. Campylobacter jejuni subtypes were closely related to MLST sequence type 21 (ST-21) and had identical porA sequences. Campylobacter coli subtypes were unrelated to isolates in the pubMLST/porA database. This feral swine population lived in close association with a 'grassfed' beef cattle herd adjacent to spinach and other leafy green row crop fields. The findings underscore the importance of protecting raw vegetable crops from faecal contamination by wild or feral animals. The study also illustrates a potential risk of Campylobacter exposure for hunters during handling and processing of wild swine meat.
Ugarte-Ruiz, M; Wassenaar, T M; Gómez-Barrero, S; Porrero, M C; Navarro-Gonzalez, N; Domínguez, L
We determined whether different methods to isolate Campylobacter (including the ISO standard 10272:2006-1) affected the genotypes detectable from poultry, at three points during slaughter: caecal content, neck skin and meat. Carcasses from 28 independent flocks were thus sampled (subset A). In addition, ten neck skin samples from four flocks, ten caecal samples from ten different flocks and ten unrelated meat samples obtained from local supermarkets were collected (subset B). Campylobacter was isolated using eight different protocols: with and without enrichment using Bolton broth, Preston broth or Campyfood broth (CFB), followed by culture on either modified Charcoal Cefoperazone Deoxycholate Agar (mCCDA) or Campyfood agar (CFA). All obtained isolates were genotyped for flaA-SVR, and over half of the isolates were also typed by MLST. The strain richness, as a measure of number of detected fla-genotypes, obtained from subset A neck skin and caecal samples was higher than that of meat samples. In half of the cases, within a flock, at least one identical fla-genotype was obtained at all three slaughter stages, suggestive of autologous contamination of carcasses. Enrichment reduced the observed richness of isolates, while CFA plates increased richness compared to mCCDA plates, irrespective of inclusion of an enrichment step. Because the isolation protocol used influences both the yield and the fla-genotype richness obtained from poultry, this variable should be taken into account when different studies are being compared.
Oyarzabal, O A; Conner, D E; Hoerr, F J
Avian species necropsied at the C. S. Roberts Veterinary Diagnostic Laboratory, Auburn, Alabama, from December 1993 until May 1994 were examined for the incidence of intestinal campylobacters. Ninety-one intestinal swabs, representing 66 separate cases and 17 different avian species, were collected and placed into Cary-Blair transport medium. Selective enrichment and culture media were used for initial isolation of Campylobacter spp. Presumptive colonies were identified as Campylobacter spp. by phase-contrast microscopy and Gram stain, and they were confirmed by serological latex agglutination. Campylobacter spp. were isolated in 18 (19.7%) of the 66 cases. From the remainder of the cases, 13 (15%) yielded presumptive colonies on Campy-Cefex agar; however, they were not confirmed serologically as Campylobacter spp. Use of Cary-Blair transport medium held in refrigeration for up to 24 days did not hinder the determination of campylobacters in intestinal samples. A variety of avian species, including chicken, emu, hawk, ostrich, and parrot, harbored commensal campylobacters and therefore should be considered potential reservoirs.
Umunnabuike, A C; Irokanulo, E A
A total of 690 adult cockroaches (Periplaneta americana (L.), the American cockroach, and Blatta orientalis (L.), the oriental cockroach) were captured alive within domestic kitchens and near poultry houses in Vom. Using selective media, their external surfaces and internal (gut) contents, after adequate decontamination of the external surfaces, were culturally examined for the presence of campylobacters. 4 isolates of Campylobacter subsp jejuni were made (0.5%); 3 from the gut contents and 1 from the external surface. Nocardia asteroides was isolated from the gut contents of a batch of ten cockroaches. The low isolation rate notwithstanding, our results suggest that cockroaches may be a potential vector of campylobacters from other sources to human food. The somewhat fortuitous isolation of Nocardia asteroides and its significance are discussed.
Characterization of Antimicrobial Susceptibility and Its Association with Virulence Genes Related to Adherence, Invasion, and Cytotoxicity in Campylobacter jejuni and Campylobacter coli Isolates from Animals, Meat, and Humans.
Lapierre, Lisette; Gatica, María A; Riquelme, Víctor; Vergara, Constanza; Yañez, José Manuel; San Martín, Betty; Sáenz, Leonardo; Vidal, Maricel; Martínez, María Cristina; Araya, Pamela; Flores, Roberto; Duery, Oscar; Vidal, Roberto
The aim of this research was to statistically analyze the association between antimicrobial susceptibility/resistance to erythromycine, gentamicin, ciprofloxacin, and tetracycline and 11 virulence genes associated with adherence, invasion, and cytotoxicity in 528 isolates of Campylobacter coli and Campylobacter jejuni obtained from retail meat and fecal samples from food-producing animals and human patients. A high percentage of Campylobacter strains were resistant to antimicrobials, specifically ciprofloxacin and tetracycline. Moreover, we observed a wide distribution of virulence genes within the analyzed strains. C. jejuni strains were more susceptible to antimicrobials, and showed greater number of virulence genes than C. coli strains. Genes related to invasion capability, such as racR, ciaB, and pldA, were associated with antimicrobial-susceptible strains in both species. The genes cdtA and dnaJ, a citotoxin unit and an adherence-related gene, respectively, were associated with antimicrobial-resistant strains in both species. In conclusion, Campylobacter strains show a statistically significant association between antimicrobial susceptibility and the presence of virulence genes.
Igimi, S; Okada, Y; Ishiwa, A; Yamasaki, M; Morisaki, N; Kubo, Y; Asakura, H; Yamamoto, S
Campylobacter is one of the most frequently diagnosed bacterial causes of human gastroenteritis in Japan and throughout the world. Resistance to quinolones in Campylobacter jejuni and C. coli isolated from humans has emerged in many countries during the past 15 years because fluoroquinolones are the drug of choice for the treatment of suspected bacterial gastroenteritis. Food contaminated with Campylobacter is the usual source of human infection; therefore, the presence of antimicrobial resistance strains in the food chain has raised concerns that the treatment of human infections will be compromised. The use of antimicrobial agents for food animals and in veterinary medicine is suspected to be correlated with an increase in quinolone-resistant strains of Campylobacter in food animals, especially in poultry products. In contrast to macrolide resistance in C. jejuni and C. coli isolated from humans showing a stable low rate, resistant Campylobacter spp. to quinolones have emerged in Japan. The paper summarizes food-borne Campylobacter infection in Japan, and the prevalence and trends of antimicrobial resistance of Campylobacter from the authors' data and other Japanese papers which reported the antimicrobial resistance of Campylobacter.
Burke, V; Robinson, J; Gracey, M; Peterson, D; Meyer, N; Haley, V
The recovery of Aeromonas spp. from the unchlorinated water supply for a Western Australian city of 21,000 people was monitored at several sampling points during a period of 1 year. Membrane filtration techniques were used to count colonies of Aeromonas spp., coliforms, and Escherichia coli in water sampled before entry to service reservoirs, during storage in service reservoirs, and in distribution systems. Aeromonas spp. were identified by subculture on blood agar with ampicillin, oxidase tests, and the use of Kaper medium and then were tested for production of enterotoxins and hemolysins. During the same period, two-thirds of all fecal specimens sent for microbiological examination were cultured on ampicillin-blood agar for Aeromonas spp. Recovery of Aeromonas spp. from water supplies at distribution points correlated with fecal isolations and continued during autumn and winter. Coliforms and E. coli were found most commonly in late summer to autumn. This pattern differs from the summer peak of Aeromonas isolations both from water and from patients with Aeromonas spp.-associated gastroenteritis in Perth, Western Australia, a city with a chlorinated domestic water supply. Of the Aeromonas strains from water, 61% were enterotoxigenic, and 64% produced hemolysins. PMID:6486783
There is no universally accepted standard method for the isolation of Campylobacter spp. and it is considered that currently available isolation media are not yet optimal for the recovery of Campylobacter spp. from a range of sample types. Almost all methods incorporate antibiotics into the isolation media to inhibit growth of other bacteria within the sample. It is established that the incorporation of such antibiotics into isolation media will inhibit the growth of some Campylobacter spp. as well as other bacteria. The results of the use of such suboptimal isolation methods are that the isolates which 'survive' the isolation procedure will be those which: (i) are able to 'out compete' the rest of the bacteria in the sample, i.e. they are able to grow faster; (ii) are resistant to the antibiotics used in the isolation media; and (iii) are randomly selected by the laboratory technician as being a 'typical'Campylobacter spp. It is clear that such a procedure is intrinsically biased and will mean that species resistant to the antibiotics used in the media will be isolated. This introduces real doubt that the bacteria isolated are truly representative of those initially found on the sample. It is also becoming clear that Campylobacter spp. are rather difficult to isolate as pure cultures and many are in fact mixtures of more than one strain. Again this introduces great uncertainty as to the prevalence and distribution of respective species from the different sample types. This is especially true when considering isolation of Campylobacter spp. causing disease in man as there is no certainty that the selected isolate is that which was responsible for disease. The incorporation of antibiotics into the isolation media not only introduces the issue of species bias but perhaps more importantly exposes the Campylobacter spp. to a cocktail of antibiotics thereby providing the potential for them to 'switch on' antibiotic resistance mechanisms. It might be argued that this has
Diker, K S; Yardimci, H
Several isolation methods and media and the effect of moisture content on colony morphology were compared for the primary isolation of Campylobacter jejuni. Of the 200 rectal swabs from cattle and sheep tested for the isolation of C. jejuni, 19.5% were positive on Butzler medium and 16.5% were positive on Skirrow medium. The transportation of samples in modified Cary-Blair medium increased the isolation rate. Of the 300 gallbladder from cattle and sheep tested for the isolation of C. jejuni, 5% were positive by direct inoculation of bile, 27% were positive by swabbing and 31% were positive by selective enrichment method. The organism produced two different type of colonies on fresh and dried media.
Jenkins, J.A.; Taylor, P.W.
Two solid bacteriologic media were compared for cultivating Aeromonas spp. from piscine sources: the Rimler-Shotts (RS) medium and a starch-glutamate-ampicillin-penicillin-based medium (SGAP-10C) used for the recovery of Aeromonas spp. from water samples. The selective and differential capacities of the media were assessed March through October 1992 by recovery rate and phenotype of 99 isolates representing 15 genera of bacteria. Recovery frequency of Aeromonas spp. (n = 62) was similar at 97% on RS and 95% on SGAP-10C. The SGAP-10C medium proved to be more specific than RS toward Aeromonas species (P ≤ 0.005). Use of SGAP-10C at 24 C for 48 hr offers a better choice for the laboratory recovery of Aeromonas spp. from clinical fish specimens.
Pearson, Bruce M; Louwen, Rogier; van Baarlen, Peter; van Vliet, Arnoud H M
CRISPR (clustered regularly interspaced palindromic repeats)-Cas (CRISPR-associated) systems are sequence-specific adaptive defenses against phages and plasmids which are widespread in prokaryotes. Here we have studied whether phylogenetic relatedness or sharing of environmental niches affects the distribution and dissemination of Type II CRISPR-Cas systems, first in 132 bacterial genomes from 15 phylogenetic classes, ranging from Proteobacteria to Actinobacteria. There was clustering of distinct Type II CRISPR-Cas systems in phylogenetically distinct genera with varying G+C%, which share environmental niches. The distribution of CRISPR-Cas within a genus was studied using a large collection of genome sequences of the closely related Campylobacter species Campylobacter jejuni (N = 3,746) and Campylobacter coli (N = 486). The Cas gene cas9 and CRISPR-repeat are almost universally present in C. jejuni genomes (98.0% positive) but relatively rare in C. coli genomes (9.6% positive). Campylobacter jejuni and agricultural C. coli isolates share the C. jejuni CRISPR-Cas system, which is closely related to, but distinct from the C. coli CRISPR-Cas system found in C. coli isolates from nonagricultural sources. Analysis of the genomic position of CRISPR-Cas insertion suggests that the C. jejuni-type CRISPR-Cas has been transferred to agricultural C. coli. Conversely, the absence of the C. coli-type CRISPR-Cas in agricultural C. coli isolates may be due to these isolates not sharing the same environmental niche, and may be affected by farm hygiene and biosecurity practices in the agricultural sector. Finally, many CRISPR spacer alleles were linked with specific multilocus sequence types, suggesting that these can assist molecular epidemiology applications for C. jejuni and C. coli.
Pearson, Bruce M.; Louwen, Rogier; van Baarlen, Peter; van Vliet, Arnoud H.M.
CRISPR (clustered regularly interspaced palindromic repeats)-Cas (CRISPR-associated) systems are sequence-specific adaptive defenses against phages and plasmids which are widespread in prokaryotes. Here we have studied whether phylogenetic relatedness or sharing of environmental niches affects the distribution and dissemination of Type II CRISPR-Cas systems, first in 132 bacterial genomes from 15 phylogenetic classes, ranging from Proteobacteria to Actinobacteria. There was clustering of distinct Type II CRISPR-Cas systems in phylogenetically distinct genera with varying G+C%, which share environmental niches. The distribution of CRISPR-Cas within a genus was studied using a large collection of genome sequences of the closely related Campylobacter species Campylobacter jejuni (N = 3,746) and Campylobacter coli (N = 486). The Cas gene cas9 and CRISPR-repeat are almost universally present in C. jejuni genomes (98.0% positive) but relatively rare in C. coli genomes (9.6% positive). Campylobacter jejuni and agricultural C. coli isolates share the C. jejuni CRISPR-Cas system, which is closely related to, but distinct from the C. coli CRISPR-Cas system found in C. coli isolates from nonagricultural sources. Analysis of the genomic position of CRISPR-Cas insertion suggests that the C. jejuni-type CRISPR-Cas has been transferred to agricultural C. coli. Conversely, the absence of the C. coli-type CRISPR-Cas in agricultural C. coli isolates may be due to these isolates not sharing the same environmental niche, and may be affected by farm hygiene and biosecurity practices in the agricultural sector. Finally, many CRISPR spacer alleles were linked with specific multilocus sequence types, suggesting that these can assist molecular epidemiology applications for C. jejuni and C. coli. PMID:26338188
Rahim, Z.; Khan, S. I.; Chopra, A. K.
Cytotoxic enterotoxin (Act) is a key virulence factor in the pathogenesis of infections caused by Aeromonas spp. The cytotoxic enterotoxin gene (act) was detected in 32 out of 69 environmental isolates of Aeromonas spp. by hybridization with the act gene probe. To evaluate the pathogenic potential of the act gene probe-positive isolates, 32 act gene probe-positive and 31 randomly selected act gene probe-negative isolates were tested for enterotoxicity in a suckling mice assay (SMA), for haemolytic activity on sheep blood agar plates, for the presence of CAMP-like factors, and for cytotoxicity in a Vero cell line. The act gene probe-positive isolates significantly differed from the toxin gene probe-negative ones with respect to enterotoxicity in the SMA (P=0.009) and haemolytic activity (P=0.005). The CAMP-haemolysin phenotype was significantly associated with the rabbit ileal loop assay (P= 0.08), Vero cell assay (P= 0.064), and haemolysin production under the microaerophilic conditions (P= 0.056) of the act gene probe-positive isolates of Aeromonas spp. These data indicated the role of Act in the pathogenesis of Aeromonas infections and that the enterotoxic potential of Aeromonas spp. could be assessed by simply performing a CAMP-haemolysin assay. PMID:15310164
Carreira, Ana Cláudia; Clemente, Lurdes; Rocha, Teresa; Tavares, Alcina; Geraldes, Margarida; Barahona, Maria José; Botelho, Ana; Cunha, Mónica V
Campylobacter is a major cause of human foodborne disease worldwide and has been associated with the consumption of contaminated poultry. The prevalence of Campylobacter species in broiler flocks from more than 200 producers widespread in mainland Portugal was assessed in 2008 in response to Commission Decision 2007/516/EC. Campylobacter isolates were obtained from 83.3% of 424 pooled cecal samples, with a higher prevalence of Campylobacter coli (61.2%) than Campylobacter jejuni (38.8%). Restriction fragment length polymorphism analysis of the flaA gene (flaA-RFLP) of 112 C. coli isolates and 67 C. jejuni isolates yielded 11 flaA-RFLP patterns with HinfI (Hunter Gaston diversity index [HGDI] = 0.62) and 48 flaA-RFLP patterns with DdeI (HGDI = 0.89), indicating a high level of genetic diversity. A wide geographic distribution of the most frequent restriction pattern was observed. MICs of five antimicrobials (ciprofloxacin, gentamicin, streptomycin, erythromycin, and tetracycline) were determined for 215 C. coli isolates and 136 C. jejuni isolates. The occurrence of non-wild-type isolates, exhibiting an acquired resistance phenotype, was higher for C. coli than C. jejuni for all antimicrobials tested. Sixty-three percent of C. jejuni and C. coli isolates were considered non-wild type based on their response to both ciprofloxacin and erythromycin, which are frequently used in the treatment of human infections. The high prevalence of antimicrobial-resistant Campylobacter strains detected supports the need for increased epidemiological surveillance and prevention in a country where large amounts of poultry meat are consumed.
Taveirne, Michael E.; Dunham, Drew T.; Perault, Andrew; Beauchamp, Jessica M.; Huynh, Steven; Parker, Craig T.
ABSTRACT Campylobacter jejuni is a leading cause of bacterially derived foodborne illness. Human illness is commonly associated with the handling and consumption of contaminated poultry products. Three C. jejuni strains were isolated from cecal contents of three different naturally colonized farm-raised chickens. The complete genomes of these three isolates are presented here. PMID:28126931
Korotkevich, Yu V
The isolates from foods were screened for sensitivity to clinically significant antibiotics to assess the actual situation related to the prevalence of the antibiotic-resistant microorganisms in food. The goal of this work was to study the phenotypic characteristics of the antibiotic susceptibility of Enterobacteriaceae and Enterococcus spp. isolated from the good quality foods, and evaluation of the prevalence of tetracycline resistance in this groups of microbial contaminants. 68 strains of Enterobacteriaceae family and Enterococcus spp. isolated from poultry and livestock meat, pasteurized dairy products, acquired in the retail in the Moscow region, were studied. The disk-diffusion method (DDM) analysis showed a rather high prevalence of bacteria that are resistant and forming resistance to broad-spectrum antibiotics: in general 38% of Enterobacteriaceae strains and 40% of Enterococcus spp., isolated from meat products were resistant to tetracycline and doxycycline, and 21 and 33% - from dairy products, respectively; 26% of milk isolates and 54% of meat isolates were resistant to ampicillin. Considering that the tetracyclines is the most frequently used in animal husbandry and veterinary, the incidence and levels of tetracycline resistance were evaluated using tests with higher sensitivity to minimum inhibitory concentration (MIC), than the DDM. It was shown that among the Enterobacteriaceae strains 26% of
Schweitzer, Nóra; Damjanova, Ivelina; Kaszanyitzky, Eva; Ursu, Krisztina; Samu, Péterné; Tóth, Adám György; Varga, János; Dán, Adám
During 2008 and 2009, within the framework of the Hungarian monitoring program of antibiotic resistance of zoonotic agents from food-producing animals, a significant number (43 strains) of Campylobacter lanienae were detected for the first time in Hungary. The isolates were genotyped using partial 16S rRNA gene sequencing and pulsed-field gel electrophoresis using three different restriction enzymes. The antimicrobial resistance of the isolates was determined by microtiter broth dilution. C. lanienae isolation was successful only from swine but not from other animal species. According to phylogenetic analysis, clustering of the isolates shows the same extensive genetic diversity as other Campylobacter species. Sequence analysis of the partial 16S rRNA gene showed that additional variations exist in variable regions Vc2 and Vc6. SmaI restriction enzyme proved to be the most efficient for pulsed-field gel electrophoresis analysis of C. lanienae. A significant tetracycline resistance (60.9%) and the presence of erythromycin-, enrofloxacin-, and multiresistant C. lanienae strains were found. Although the pathogenic potential of C. lanienae in humans is currently unknown, this study demonstrates that C. lanieanae is common in pigs in the country, provides further details on the genotypic and phenotypic properties of C. lanienae, and offers a genotyping method for use in source tracing.
Aarts, H J; van Lith, L A; Jacobs-Reitsma, W F
Thirty-four Campylobacter jejuni or coli strains, isolated from various livestock and darkling beetles from two Dutch poultry farms during different broiler production cycles, were subjected to Penner serotyping and polymerase chain reaction (PCR) fingerprint analysis. Ten different Penner serotypes were determined in the isolates. Visual scoring of the PCR fingerprints resulted in 14 clearly different profiles. Some strains with identical Penner serotypes exhibited different PCR fingerprints and conversely strains with different serotypes produced identical PCR fingerprints. Discrepancies between Penner serotyping and PCR fingerprinting were most obvious between isolates from different animal sources. Indications for the occurrence of genomic rearrangements were found. The inconsistency between serotyping and fingerprinting of Campylobacter strains suggests that conventional typing methods should be used in combination with fingerprinting if the epidemiological factors that contribute to Campylobacter colonization of live chickens are to be assessed reliably.
Signorini, M L; Zbrun, M V; Romero-Scharpen, A; Olivero, C; Bongiovanni, F; Soto, L P; Frizzo, L S; Rosmini, M R
Here, we developed a quantitative risk assessment for thermophilic Campylobacter spp. related to the consumption of salad prepared alongside broiler meat. The assessment considered initial contamination levels, cross-contamination and decontamination events during the broiler slaughter process and distribution, and storage and consumption patterns in Argentina and other Latin American countries. The model predicted an infection risk of 3.32×10(-4) per serving. This estimation was variable according to the dose-response model used. Considering the number of chickens slaughtered annually in Argentina, the estimated number of people who could suffer campylobacteriosis related to poultry meat consumption was, on average, 484,304. The risk of human campylobacteriosis was most sensitive to the probability of infection from a Campylobacter (r=0.72), the number of Campylobacter spp. per serving (r=0.40), the frequency of washing the cutting board (r=-0.31), the preparation of raw poultry before salad using the same cutting board (r=0.14), and the frequency of hand washing (r=-0.14). The most sensitive stages of the process identified through the risk assessment can be used as a basis for measures of risk management. Public campaigns on hygiene habits during food preparation at home should focus on the importance of washing the cutting board before preparing raw and ready-to-eat foods and of washing the hands during food preparation.
Serichantalergs, Oralak; Jensen, L B; Pitarangsi, C; Mason, C J; Dalsgaard, A
A total of 495 Campylobacterjejuni and 122 C. coli isolated from Thai children were screened for macrolide (erythromycin and azithromycin) resistance by disk diffusion assay. Minimum inhibitory concentrations for erythromycin, azithromycin, nalidixic acid, ciprofloxacin, tetracycline, streptomycin, gentamicin and chloramphenicol were further determined for these macrolide-resistant Campylobacter isolates. Presence of known point mutations resulting in reduced susceptibility to macrolides was investigated by PCR and DNA sequencing. Seventeen percent (23/122) of C. coli and 2.4% (12/495) of C. jejuni isolates were resistant to macrolides. By sequencing domain V of the 23S ribosomal DNA from all 35 macrolide-resistant isolates, a known point mutation of 23S rRNA associated with reduced susceptibility to macrolides was detected in all isolates except one. Among the macrolide-resistant isolates, all were multiply resistant to nalidixic acid and ciprofloxacin, of which the latter is the preferred antimicrobial used for diarrheal treatment in Thailand. Furthermore, most macrolide-resistant isolates were also resistant to tetracycline and streptomycin. The spread of macrolide and quinolone resistant Campylobacter should be monitored closely in Thailand and elsewhere as these antimicrobials are preferred drugs for treatment of diarrhea.
Rodríguez-Martínez, Sara; Blanky, Marina; Friedler, Eran; Halpern, Malka
Legionella, an opportunistic human pathogen whose natural environment is water, is transmitted to humans through inhalation of contaminated aerosols. Legionella has been isolated from a high diversity of water types. Due its importance as a pathogen, two ISO protocols have been developed for its monitoring. However, these two protocols are not suitable for analyzing Legionella in greywater (GW). GW is domestic wastewater excluding the inputs from toilets and kitchen. It can serve as an alternative water source, mainly for toilet flushing and garden irrigation; both producing aerosols that can cause a risk for Legionella infection. Hence, before reuse, GW has to be treated and its quality needs to be monitored. The difficulty of Legionella isolation from GW strives in the very high load of contaminant bacteria. Here we describe a modification of the ISO protocol 11731:1998 that enables the isolation and quantification of Legionella from GW samples. The following modifications were made:•To enable isolation of Legionella from greywater, a pre-filtration step that removes coarse matter is recommended.•Legionella can be isolated after a combined acid-thermic treatment that eliminates the high load of contaminant bacteria in the sample.
Rodríguez-Martínez, Sara; Blanky, Marina; Friedler, Eran; Halpern, Malka
Legionella, an opportunistic human pathogen whose natural environment is water, is transmitted to humans through inhalation of contaminated aerosols. Legionella has been isolated from a high diversity of water types. Due its importance as a pathogen, two ISO protocols have been developed for its monitoring. However, these two protocols are not suitable for analyzing Legionella in greywater (GW). GW is domestic wastewater excluding the inputs from toilets and kitchen. It can serve as an alternative water source, mainly for toilet flushing and garden irrigation; both producing aerosols that can cause a risk for Legionella infection. Hence, before reuse, GW has to be treated and its quality needs to be monitored. The difficulty of Legionella isolation from GW strives in the very high load of contaminant bacteria. Here we describe a modification of the ISO protocol 11731:1998 that enables the isolation and quantification of Legionella from GW samples. The following modifications were made:•To enable isolation of Legionella from greywater, a pre-filtration step that removes coarse matter is recommended.•Legionella can be isolated after a combined acid-thermic treatment that eliminates the high load of contaminant bacteria in the sample. PMID:26740925
Banowary, Banya; Dang, Van Tuan; Sarker, Subir; Connolly, Joanne H; Chenu, Jeremy; Groves, Peter; Ayton, Michelle; Raidal, Shane; Devi, Aruna; Vanniasinkam, Thiru; Ghorashi, Seyed A
Campylobacter spp. are important causes of bacterial gastroenteritis in humans in developed countries. Among Campylobacter spp. Campylobacter jejuni (C. jejuni) and C. coli are the most common causes of human infection. In this study, a multiplex PCR (mPCR) and high resolution melt (HRM) curve analysis were optimized for simultaneous detection and differentiation of C. jejuni and C. coli isolates. A segment of the hippuricase gene (hipO) of C. jejuni and putative aspartokinase (asp) gene of C. coli were amplified from 26 Campylobacter isolates and amplicons were subjected to HRM curve analysis. The mPCR-HRM was able to differentiate between C. jejuni and C. coli species. All DNA amplicons generated by mPCR were sequenced. Analysis of the nucleotide sequences from each isolate revealed that the HRM curves were correlated with the nucleotide sequences of the amplicons. Minor variation in melting point temperatures of C. coli or C. jejuni isolates was also observed and enabled some intraspecies differentiation between C. coli and/or C. jejuni isolates. The potential of PCR-HRM curve analysis for the detection and speciation of Campylobacter in additional human clinical specimens and chicken swab samples was also confirmed. The sensitivity and specificity of the test were found to be 100% and 92%, respectively. The results indicated that mPCR followed by HRM curve analysis provides a rapid (8 hours) technique for differentiation between C. jejuni and C. coli isolates.
El-Adawy, Hosny; Hotzel, Helmut; Düpre, Sabine; Tomaso, Herbert; Neubauer, Heinrich; Hafez, Hafez M
The emergence of antimicrobial resistance among Campylobacter isolates recovered from turkeys has increased dramatically. Monitoring the progress of this resistance becomes a growing public health issue. The aim of the present study was to provide information of the current status of antibiotic resistance patterns in Campylobacter jejuni from turkeys. Seventy-six C. jejuni isolates were recovered from 67 epidemiologically unrelated meat turkey flocks in different regions of Germany in 2010 and 2011. The isolates were typed by flaA genotyping and were investigated for antimicrobial susceptibility against 12 antibiotics by using a broth microdilution test as well as testing the genetic determination of ciprofloxacin, tetracycline, and erythromycin resistance. All isolates (n = 76) were sensitive to gentamicin and chloramphenicol. The numbers of isolates that were sensitive to streptomycin, erythromycin, neomycin, and amoxicillin were 69 (90.8%), 61 (80.2%), 58 (76.4%), and 44 (57.9%), respectively. Only one isolate was sensitive to all tested antibiotics. The emergence of a high resistance rate and multidrug resistance to three or more classes of antimicrobial agents were observed. The resistance against sulphamethoxazole/trimethoprim, metronidazole, ciprofloxacin, naladixic acid, and tetracycline was 58 (76.3%), 58 (76.3%), 53 (69.7%), 51 (67.1%), and 42 (55.3%), respectively. None of the isolates was resistant to all antibiotics. Multidrug resistance to three or more classes of antimicrobial agents was found and ranged from 3.9% to 40.8%. Replacement of the Thr-86-->Ile in gyrA gene and detection of the tet(O) gene were the main resistance mechanisms for fluoroquinolones and tetracycline, respectively, while the lack of mutation in position 2074 and 2075 on the 23S rRNA gene was responsible for macrolide resistance. The phenotypic and genotypic resistance profiles were compatible in the case of ciprofloxacin and tetracycline but were not completely congruent with
Haruna, M; Sasaki, Y; Murakami, M; Ikeda, A; Kusukawa, M; Tsujiyama, Y; Ito, K; Asai, T; Yamada, Y
Campylobacter was isolated from 67 (47.2%) of 142 broiler flocks between September 2009 and February 2010. The prevalence of Campylobacter in broiler flocks was significantly lower during January and February than it was from September to December. Campylobacter colonization was more common in flocks that were not provided with a disinfected water supply, which was consistent with the findings of a previous study. The prevalence of antimicrobial drug-resistant Campylobacter spp. was investigated, and the minimum inhibitory concentrations of eight antimicrobial agents were determined for 122 Campylobacter jejuni isolates and 46 Campylobacter coli isolates from broiler flocks between 2007 and 2010. In this study, 29.5% (36/122) of C. jejuni isolates and 41.3% (19/46) of C. coli isolates were resistant to enrofloxacin (ERFX), whereas all isolates were susceptible to erythromycin. Furthermore, the ERFX-resistant isolates were tested for susceptibility to other classes of antimicrobial agents, and 55.6% (20/36) of ERFX-resistant C. jejuni isolates and 47.4% (9/19) of ERFX-resistant C. coli isolates were resistant to at least one of aminobenzyl penicillin, dihydrostreptomycin and oxytetracycline. To avoid an impact of antimicrobial drug-resistant Campylobacter spp. on the efficacy of antimicrobial treatment for human campylobacteriosis, prudent use of antimicrobial agents is a requisite. The use of antimicrobial agents should be accompanied by various approaches such as prevention of Campylobacter colonization in broiler flocks with the aim of lowering the occurrence of Campylobacter infection in humans.
Cook, Angela; Reid-Smith, Richard J; Irwin, Rebecca J; McEwen, Scott A; Young, Virginia; Ribble, Carl
This study estimated the prevalence of Salmonella, Campylobacter, and Escherichia coli isolates in fresh retail grain-fed veal obtained in Ontario, Canada. The prevalence and antimicrobial resistance patterns were examined for points of public health significance. Veal samples (n = 528) were collected from February 2003 through May 2004. Twenty-one Salmonella isolates were recovered from 18 (4%) of 438 samples and underwent antimicrobial susceptibility testing. Resistance to one or more antimicrobials was found in 6 (29%) of 21 Salmonella isolates; 5 (24%) of 21 isolates were resistant to five or more antimicrobials. No resistance to antimicrobials of very high human health importance was observed. Ampicillin-chloramphenicolstreptomycin-sulfamethoxazole-tetracycline resistance was found in 5 (3%) of 21 Salmonella isolates. Campylobacter isolates were recovered from 5 (1%) of 438 samples; 6 isolates underwent antimicrobial susceptibility testing. Resistance to one or more antimicrobials was documented in 3 (50%) of 6 Campylobacter isolates. No Campylobacter isolates were resistant to five or more antimicrobials or category I antimicrobials. E. coli isolates were recovered from 387 (88%) of 438 samples; 1,258 isolates underwent antimicrobial susceptibility testing. Resistance to one or more antimicrobials was found in 678 (54%) of 1,258 E. coli isolates; 128 (10%) of 1,258 were resistant to five or more antimicrobials. Five (0.4%) and 7 (0.6%) of 1,258 E. coli isolates were resistant to ceftiofur and ceftriaxone, respectively, while 34 (3%) of 1,258 were resistant to nalidixic acid. Ciprofloxacin resistance was not detected. There were 101 different resistance patterns observed among E. coli isolates; resistance to tetracycline alone (12.7%, 161 of 1,258) was most frequently observed. This study provides baseline prevalence and antimicrobial resistance data and highlights potential public health concerns.
Hart, W S; Heuzenroeder, M W; Barton, M D
The major influences on the amplification and spread of antibiotic-resistant bacteria are the therapeutic use of antibiotics in human medicine and their use in livestock for therapy, prophylaxis and growth promotion. The use of veterinary antibiotics has many benefits to the livestock industries ensuring animal health and welfare, but use at subtherapeutic levels also exerts great selective pressure on emergence of resistant bacteria. The possible effect on human health is a problem of current debate. This study involved sampling pig carcasses, pig meat and assessing the level of resistance in zoonotic enteric bacteria of concern to human health. In South Australian pigs, thermophilic Campylobacter species showed widespread resistance (60-100%) to tylosin, erythromycin, lincomycin, ampicillin and tetracycline. No resistance was seen to ciprofloxacin. The enterococci demonstrated little resistance (0-30%) to vancomycin or virginiamycin, but the overall results from the antibiotic sensitivity testing of the enterococci have demonstrated how widespread their resistance has become. Escherichia coli strains showed widespread resistance to tetracycline and moderately common resistance (30-60%) to ampicillin and sulphadiazine. Resistance to more than one antibiotic was common. Pigs from New South Wales were also sampled and differences in resistance patterns were noted, perhaps reflecting different antibiotic use regimens in that state.
Pérez-Boto, D; Herrera-León, S; García-Peña, F J; Abad-Moreno, J C; Echeita, M A
The aim of this study was to investigate the resistance mechanisms of quinolones, macrolides and tetracycline in campylobacter isolates from grandparent and parent broiler breeders in Spain. Twenty-six isolates were investigated for quinolone resistance, three isolates for macrolide resistance and 39 for tetracycline resistance. All of the quinolone-resistant isolates possessed the mutation Thr86Ile in the quinolone resistance-determining region of gyrA and one isolate possessed the mutation Pro104Ser. Only one Campylobacter coli population (defined by restriction fragment length polymorphism-polymerase chain reaction of flaA and pulsed field gel electrophoresis) was resistant to erythromycin, and the mutation A2075G (23S rDNA) was responsible for macrolide resistance. The tetO gene was found in all of the tetracycline-resistant isolates. Twenty-two out of the 39 isolates investigated by Southern blot possessed chromosomic location of tetO and 17 were located on plasmids. Most of the plasmids with tetO were of around 60 kb and conjugation was demonstrated in a selection of them. In conclusion, we showed that Thr86Ile is highly prevalent in quinolone-resistant isolates as well as mutation A2075G in macrolide-resistant isolates of poultry origin. More variability was found for tetO. The possibility of horizontal transmission of tetO among campylobacter isolates is also an issue of concern in public health.
Wiedmann, Martin; Weilmeier, Denise; Dineen, Sean S.; Ralyea, Robert; Boor, Kathryn J.
Putative Pseudomonas spp. isolated predominantly from raw and processed milk were characterized by automated ribotyping and by biochemical reactions. Isolates were biochemically profiled using the Biolog system and API 20 NE and by determining the production of proteases, lipases, and lecithinases for each isolate. Isolates grouped into five coherent clusters, predominated by the species P. putida (cluster A), P. fluorescens (cluster B), P. fragi (as identified by Biolog) or P. fluorescens (as identified by API 20 NE) (cluster C), P. fragi (as identified by Biolog) or P. putida (as identified by API 20 NE) (cluster D), and P. fluorescens (cluster E). Isolates within each cluster also displayed similar enzyme activities. Isolates in clusters A, C, and D were generally negative for all three enzyme activities; isolates in cluster B were predominantly positive for all three enzyme activities; and isolates in cluster E were negative for lecithinase but predominantly positive for protease and lipase activities. Thus, only isolates from clusters B and E produced enzyme activities associated with dairy product flavor defects. Thirty-eight ribogroups were differentiated among the 70 isolates. Ribotyping was highly discriminatory for dairy Pseudomonas isolates, with a Simpson's index of discrimination of 0.955. Isolates of the same ribotype were never classified into different clusters, and ribotypes within a given cluster generally showed similar ribotype patterns; thus, specific ribotype fragments may be useful markers for tracking the sources of pseudomonads in dairy production systems. Our results suggest that ribogroups are generally homogeneous with respect to nomenspecies and biovars, confirming the identification potential of ribotyping for Pseudomonas spp. PMID:10788386
Detecting and enumerating Campylobacter from poultry samples can be difficult without highly selective media because of competing microflora. We have found that adding 0.1 µg/ mL of Triclosan, an antibacterial agent, to Bolton enrichment broth prevents overgrowth of non-Campylobacter bacteria and si...
Bernal, Johan F.; Donado-Godoy, Pilar; Arévalo, Alejandra; Duarte, Carolina; Realpe, María E.; Díaz, Paula L.; Gómez, Yolanda; Rodríguez, Fernando; Agarwala, Richa; Landsman, David
Campylobacter coli is considered one of the main causes of food-borne illness worldwide. We report here the whole-genome sequence of multidrug-resistant Campylobacter coli strain COL B1-266, isolated from the Colombian poultry chain. The genome sequences encode genes for a variety of antimicrobial resistance genes, including aminoglycosides, β-lactams, lincosamides, fluoroquinolones, and tetracyclines. PMID:26988047
Campylobacter jejuni is a major cause of human foodborne illness worldwide with contaminated poultry products serving as a main source of human infection. C. jejuni strain MTVDSCj20 was isolated from the cecal contents of a farm-raised chicken naturally colonized with Campylobacter. The complete,...
Bernal, Johan F; Donado-Godoy, Pilar; Arévalo, Alejandra; Duarte, Carolina; Realpe, María E; Díaz, Paula L; Gómez, Yolanda; Rodríguez, Fernando; Agarwala, Richa; Landsman, David; Mariño-Ramírez, Leonardo
Campylobacter coli is considered one of the main causes of food-borne illness worldwide. We report here the whole-genome sequence of multidrug-resistant Campylobacter coli strain COL B1-266, isolated from the Colombian poultry chain. The genome sequences encode genes for a variety of antimicrobial resistance genes, including aminoglycosides, β-lactams, lincosamides, fluoroquinolones, and tetracyclines.
We report the isolation of Campylobacter species from the same population of feral swine that was investigated in San Benito County, California during the 2006 spinach-related Escherichia coli O157:H7 outbreak. This is the first survey of Campylobacter in a free-ranging feral swine population in the...
Crawshaw, Tim R; Chanter, Jeremy I; Young, Stuart C L; Cawthraw, Shaun; Whatmore, Adrian M; Koylass, Mark S; Vidal, Ana B; Salguero, Francisco J; Irvine, Richard M
The condition known as spotty liver disease or spotty liver syndrome can cause significant mortality in free range laying hen flocks. It has been described in Europe and Australia but the aetiology has not been established. There are similarities between spotty liver disease and avian vibrionic hepatitis, a condition which was reported in the 1950s. A Vibrio-like organism was suspected to be the cause of avian vibrionic hepatitis, although this organism was never fully characterised. We report the isolation of a novel Campylobacter from five separate outbreaks of spotty liver disease. The conditions required for culture, the growth characteristics, electron microscopical morphology and results of the phenotypic tests used in the identification of this novel Campylobacter sp. are described. The novel Campylobacter is slow growing and fastidious and does not grow on media routinely used for isolating Campylobacter sp. The morphology is typical for a Campylobacter sp. and phenotypic tests and a duplex real time PCR test differentiate the novel Campylobacter from other members of the genus. 16S rRNA analysis of 19 isolates showed an identical sequence which appears to represent a hitherto unknown sub lineage within the genus Campylobacter. Experimental intraperitoneal infection of four week old SPF chickens produced microscopic liver pathology indistinguishable from natural disease and the novel Campylobacter was recovered from the experimentally infected chicks. The isolates described appear to be a possible causal organism for spotty liver disease.
Lajhar, S A; Jennison, A V; Patel, B; Duffy, L L
Campylobacter jejuni is responsible for most foodborne bacterial infections worldwide including Australia. The aim of this study was to investigate a combination of typing methods in the characterization of C. jejuni isolated from clinical diarrhoeal samples (n = 20) and chicken meat (n = 26) in order to identify the source of infection and rank isolates based on their relative risk to humans. Sequencing of the flaA short variable region demonstrated that 86% of clinical isolates had genotypes that were also found in chicken meat. A polymerase chain reaction binary typing system identified 27 different codes based on the presence or absence of genes that have been reported to be associated with various aspects of C. jejuni pathogenicity, indicating that not all isolates may be of equal risk to human health. The lipooligosaccharide (LOS) of the C. jejuni isolates was classified into six classes (A, B, C, E, F, H) with 10·4% remaining unclassified. The majority (72·7%) of clinical isolates possessed sialylated LOS classes. Sialylated LOS classes were also detected in chicken isolates (80·7%). Antimicrobial tests indicated a low level of resistance, with no phenotypic resistance found to most antibiotics tested. A combination of typing approaches was useful to assign isolates to a source of infection and assess their risk to humans.
Poultry products serve as the main source of Campylobacter jejuni subsp. jejuni (Cjj) infections in humans. Cjj infections are a leading cause of foodborne gastroenteritis and are a prevalent antecedent to Guillain-Barré syndrome (GBS). This study describes the genome of Cjj HS:19 strain RM1285 isol...
Campylobacter jejuni is a leading cause of bacterially derived foodborne illness worldwide. Human illness is commonly associated with handling and consumption of contaminated poultry products. Three C. jejuni strains were isolated from cecal contents of three different naturally colonized, farm rais...
Serraino, A; Florio, D; Giacometti, F; Piva, S; Mion, D; Zanoni, R G
The objectives of this study were to investigate the presence of Campylobacter spp. and Arcobacter spp. in dairy herds authorized for the production and sale of raw milk and in a water buffalo dairy farm, and to test the antimicrobial susceptibility of the isolates. A total of 196 in-line milk filters were collected from 14 dairy farms (13 bovine and 1 water buffalo) for detection of Campylobacter spp. and Arcobacter spp. by microbiological culture. For each farm investigated, 1 isolate for each Campylobacter and Arcobacter species isolated was tested using the Etest method (AB Biodisk, Solna, Sweden) to evaluate the susceptibility to ciprofloxacin, tetracycline, chloramphenicol, ampicillin, erythromycin, and gentamicin. A total of 52 isolates were detected in 49 milk filters in 12 farms (85.7%) out of 14 and the isolates were identified as Campylobacter jejuni (6), Campylobacter hyointestinalis ssp. hyointestinalis (8), Campylobacter concisus (1), Campylobacter fetus ssp. fetus (1), Arcobacter butzleri (22), and Arcobacter cryaerophilus (14). The small number of isolates tested for antimicrobial susceptibility precludes any epidemiological consideration but highlights that all Campylobacter isolates were susceptible to macrolides, which are the first-choice drugs for the treatment of campylobacteriosis, and that resistance to fluoroquinolones and tetracycline was detected; for Arcobacter isolates, resistance to ampicillin and chloramphenicol was detected. The sale of raw milk for human consumption by self-service automatic vending machines has been allowed in Italy since 2004 and the presence of C. jejuni in in-line milk filters confirms that raw milk consumption is a significant risk factor for human infection. The high occurrence of emerging Campylobacter spp. and Arcobacter spp. discovered in dairy farms authorized for production and sale of raw milk represents an emerging hazard for human health.
Grinberg, Alex; Midwinter, Anne C.; Marshall, Jonathan C.; Collins-Emerson, Julie M.; French, Nigel P.
ABSTRACT Campylobacteriosis is one of the most important foodborne diseases worldwide and a significant health burden in New Zealand. Campylobacter jejuni is the predominant species worldwide, accounting for approximately 90% of human cases, followed by Campylobacter coli. Most studies in New Zealand have focused on C. jejuni; hence, the impact of C. coli strains on human health is not well understood. The aim of this study was to genotype C. coli isolates collected in the Manawatu region of New Zealand from clinical cases, fresh poultry meat, ruminant feces, and environmental water sources, between 2005 and 2014, to study their population structure and estimate the contribution of each source to the burden of human disease. Campylobacter isolates were identified by PCR and typed by multilocus sequence typing. C. coli accounted for 2.9% (n = 47/1,601) of Campylobacter isolates from human clinical cases, 9.6% (n = 108/1,123) from poultry, 13.4% (n = 49/364) from ruminants, and 6.4% (n = 11/171) from water. Molecular subtyping revealed 27 different sequence types (STs), of which 18 belonged to clonal complex ST-828. ST-1581 was the most prevalent C. coli sequence type isolated from both human cases (n = 12/47) and poultry (n = 44/110). When classified using cladistics, all sequence types belonged to clade 1 except ST-7774, which belonged to clade 2. ST-854, ST-1590, and ST-4009 were isolated only from human cases and fresh poultry, while ST-3232 was isolated only from human cases and ruminant sources. Modeling indicated ruminants and poultry as the main sources of C. coli human infection. IMPORTANCE We performed a molecular epidemiological study of Campylobacter coli infection in New Zealand, one of few such studies globally. This study analyzed the population genetic structure of the bacterium and included a probabilistic source attribution model covering different animal and water sources. The results are discussed in a global context. PMID:27208097
Viator, Ryan; Alles, Susan; Le, Quynh-Nhi; Hosking, Edan; Biswas, Preetha; Zhang, Lei; Tolan, Jerry; Meister, Evan; Tovar, Eric; Pinkava, Lisa; Mozola, Mark; Rice, Jennifer; Chen, Yi; Odumeru, Joseph; Ziemer, Wayne
A performance validation of the ANSR® for Campylobacter method was conducted in selected matrixes. This assay used selective nicking enzyme amplification technology to amplify target genes. Samples were enriched for 20 to 24 h and then lysed. The assay was completed within 50 min using real-time detection in a combination incubator/fluorescence detector and software. When 50 distinct strains of Campylobacter jejuni, C. lari, or C. coli were tested for inclusivity, all 50 strains produced positive results. In exclusivity testing, 31 strains of related organisms, including seven nontarget Campylobacter strains and other common species, were evaluated. All 31 species generated negative ANSR assay results, including the nontarget Campylobacter strains. The ANSR for Campylobacter method was compared to the U.S. Department of Agriculture, Food Safety and Inspection Service Microbiology Laboratory Guidebook reference method using naturally contaminated chicken carcass rinse or turkey carcass sponge samples. ANSR method performance was not statistically different from the reference method using two different enrichment options. Equivalent results were observed at both time points (20 and 24 h) and in both atmospheres (microaerobic and aerobic) to reference methods. Method performance with chicken carcass rinse was confirmed in an independent laboratory study. Additionally, in robustness testing, small, deliberate changes to the assay parameters minimally affected ANSR method performance. Finally, accelerated stability results from three independently manufactured lots supported a shelf life of 6 months when stored at 4°C. The ANSR assay offered greater efficiency and flexibility when compared to the reference method with a 20-24 h single-step enrichment in a microaerobic or an aerobic atmosphere.
Fortes-Gabriel, Elsa; Carreira, Teresa; Vieira, Maria Luísa
Rodents play an important role in the transmission of pathogenic Leptospira spp. However, in Angola, neither the natural reservoirs of these spirochetes nor leptospirosis diagnosis has been considered. Regarding this gap, we captured rodents in Luanda and Huambo provinces to identify circulating Leptospira spp. Rodent kidney tissue was cultured and DNA amplified and sequenced. Culture isolates were evaluated for pathogenic status and typing with rabbit antisera; polymerase chain reaction (PCR) and sequencing were also performed. A total of 37 rodents were captured: Rattus rattus (15, 40.5%), Rattus norvegicus (9, 24.3%), and Mus musculus (13, 35.2%). Leptospiral DNA was amplified in eight (21.6%) kidney samples. From the cultures, we obtained four (10.8%) Leptospira isolates belonging to the Icterohaemorrhagiae and Ballum serogroups of Leptospira interrogans and Leptospira borgpetersenii genospecies, respectively. This study provides information about circulating leptospires spread by rats and mice in Angola. PMID:26928840
Fortes-Gabriel, Elsa; Carreira, Teresa; Vieira, Maria Luísa
Rodents play an important role in the transmission of pathogenic Leptospira spp. However, in Angola, neither the natural reservoirs of these spirochetes nor leptospirosis diagnosis has been considered. Regarding this gap, we captured rodents in Luanda and Huambo provinces to identify circulating Leptospira spp. Rodent kidney tissue was cultured and DNA amplified and sequenced. Culture isolates were evaluated for pathogenic status and typing with rabbit antisera; polymerase chain reaction (PCR) and sequencing were also performed. A total of 37 rodents were captured: Rattus rattus (15, 40.5%), Rattus norvegicus (9, 24.3%), and Mus musculus (13, 35.2%). Leptospiral DNA was amplified in eight (21.6%) kidney samples. From the cultures, we obtained four (10.8%) Leptospira isolates belonging to the Icterohaemorrhagiae and Ballum serogroups of Leptospira interrogans and Leptospira borgpetersenii genospecies, respectively. This study provides information about circulating leptospires spread by rats and mice in Angola.
Bialvaei, Abed Zahedi; Kafil, Hossein Samadi; Asgharzadeh, Mohammad; Aghazadeh, Mohammad; Yousefi, Mehdi
This study was conducted in Iran in order to assess the distribution of CTX-M type ESBLs producing Enterobacteriaceae. From January 2012 to December 2013, totally 198 E. coli, 139 Klebsiella spp, 54 Salmonella spp and 52 Shigella spp from seven hospitals of six provinces in Iran were screened for resistance to extended-spectrum cephalosporins. After identification and susceptibility testing, isolates presenting multiple-drug resistance (MDR) were evaluated for ESBL production by the disk combination method and by Etest using (cefotaxime and cefotaxime plus clavulanic acid). All isolates were also screened for blaCTX-M using conventional PCR. A total of 42.92%, 33.81%, 14.81% and 7.69% of the E. coli, Klebsiella spp, Salmonella spp and Shigella spp isolates were MDR, respectively. The presence of CTX-M enzyme among ESBL-producing isolates was 85.18%, 77.7%, 50%, and 66.7%, in E. coli, Klebsiella spp, Salmonella spp and Shigella spp respectively. The overall presence of CTX-M genes in Enterobacteriaceae was 15.4% and among the resistant isolates was 47.6%. This study indicated that resistance to β-lactams mediated by CTX-M enzymes in Iran had similar pattern as in other parts of the world. In order to control the spread of resistance, comprehensive studies and programs are needed.
Miller, William G.; Yee, Emma; Chapman, Mary H.; Smith, Timothy P.L.; Bono, James L.; Huynh, Steven; Parker, Craig T.; Vandamme, Peter; Luong, Khai; Korlach, Jonas
The Campylobacter lari group is a phylogenetic clade within the epsilon subdivision of the Proteobacteria and is part of the thermotolerant Campylobacter spp., a division within the genus that includes the human pathogen Campylobacter jejuni. The C. lari group is currently composed of five species (C. lari, Campylobacter insulaenigrae, Campylobacter volucris, Campylobacter subantarcticus, and Campylobacter peloridis), as well as a group of strains termed the urease-positive thermophilic Campylobacter (UPTC) and other C. lari-like strains. Here we present the complete genome sequences of 11 C. lari group strains, including the five C. lari group species, four UPTC strains, and a lari-like strain isolated in this study. The genome of C. lari subsp. lari strain RM2100 was described previously. Analysis of the C. lari group genomes indicates that this group is highly related at the genome level. Furthermore, these genomes are strongly syntenic with minor rearrangements occurring only in 4 of the 12 genomes studied. The C. lari group can be bifurcated, based on the flagella and flagellar modification genes. Genomic analysis of the UPTC strains indicated that these organisms are variable but highly similar, closely related to but distinct from C. lari. Additionally, the C. lari group contains multiple genes encoding hemagglutination domain proteins, which are either contingency genes or linked to conserved contingency genes. Many of the features identified in strain RM2100, such as major deficiencies in amino acid biosynthesis and energy metabolism, are conserved across all 12 genomes, suggesting that these common features may play a role in the association of the C. lari group with coastal environments and watersheds. PMID:25381664
Thibodeau, Alexandre; Fravalo, Philippe; Garneau, Philippe; Masson, Luke; Laurent-Lewandowski, Sylvette; Quessy, Sylvain; Harel, Josée; Letellier, Ann
Campylobacter jejuni is an important worldwide foodborne pathogen commonly found as a commensal organism in poultry that can reach high numbers within the gut after colonization. Although information regarding some genes involved in colonization is available, little is known about their distribution in strains isolated specifically from chickens and whether there is a linkage between antimicrobial resistance (AMR) and colonization genes. To assess the distribution and relevance of genes associated with chicken colonization and AMR, a C. jejuni microarray was created to detect 254 genes of interest in colonization and AMR including variants. DNA derived from chicken-specific Campylobacter isolates collected in 2003 (n=29) and 2008 (n=28) was hybridized to the microarray and compared. Hybridization results showed variable colonization-associated gene presence. Acquired AMR genes were low in prevalence whereas chemotaxis receptors, arsenic resistance genes, as well as genes from the cell envelope and flagella functional groups were highly variable in their presence. Strains clustered into two groups, each linked to different control strains, 81116 and NCTC11168. Clustering was found to be independent of collection time. We also show that AMR weakly associated with the CJ0628 and arsR genes. Although other studies have implicated numerous genes associated with C. jejuni chicken colonization, our data on chicken-specific isolates suggest the opposite. The enormous variability in presumed colonization gene prevalence in our chicken isolates suggests that many are of lesser importance than previously thought. Alternatively, this also suggests that combinations of genes may be required for natural colonization of chicken intestines.
Kamei, Kazumasa; Hatanaka, Noritoshi; Asakura, Masahiro; Somroop, Srinuan; Samosornsuk, Worada; Hinenoya, Atsushi; Misawa, Naoaki; Nakagawa, Shinsaku
Campylobacter hyointestinalis isolated from swine with proliferative enteritis often is considered to be pathogenic. While the precise virulence mechanisms of this species remain unclear, we have recently identified a cytolethal distending toxin (cdt) gene cluster in C. hyointestinalis isolated from a patient with diarrhea (W. Samosornsuk et al., J Med Microbiol, 27 July 2015, http://dx.doi.org/10.1099/jmm.0.000145). However, the sequences of the cdt genes in C. hyointestinalis were found to be significantly different and the gene products are immunologically distinct from those of other Campylobacter species. In this study, we demonstrate the presence of a second variant of the cdt gene cluster in C. hyointestinalis, designated cdt-II, while the former is named cdt-I. Sequencing of the cdt-II gene cluster and deduced amino acid sequences revealed that homologies between the subunits CdtA, CdtB, and CdtC of ChCDT-I and ChCDT-II are 25.0, 56.0, and 24.8%, respectively. Furthermore, the CdtB subunit of ChCDT-II was found to be immunologically unrelated to that of ChCDT-I by Ouchterlony double gel diffusion test. Recombinant ChCDT-II also induced cell distention and death of HeLa cells by blocking the cell cycle at G2/M phase. Interestingly, the cdt-II genes were detected in all 23 animal isolates and in 1 human isolate of C. hyointestinalis, and 21 of these strains carried both cdt-I and cdt-II gene clusters. Altogether, our results indicate that ChCDT-II is an important virulence factor of C. hyointestinalis in animals. PMID:26283337
Olkkola, S; Nykäsenoja, S; Raulo, S; Llarena, A-K; Kovanen, S; Kivistö, R; Myllyniemi, A-L; Hänninen, M-L
Antimicrobial susceptibility was determined for 805 domestic Campylobacter jejuni isolates obtained from broilers (n = 459), bovines (n = 120), human patients (n = 95), natural waters (n = 80), wild birds (n = 35) and zoo animals/enclosures (n = 16) with known multilocus sequence types (MLST) for 450 isolates. The minimum inhibitory concentration (MIC) values for erythromycin, tetracycline, streptomycin, gentamicin and the quinolones ciprofloxacin and nalidixic acid were determined with the VetMIC method. MICs were compared with MLST types to find possible associations between sequence type and resistance. The proportions of resistant isolates were 5% (broilers), 6.3% (natural waters), 11.4% (wild birds), 11.6% (human patients), 16.7% (bovines) and 31.3% (zoo). The most common resistance among the human and bovine isolates was quinolone resistance alone while resistance to streptomycin alone was most often detected among the broiler isolates and tetracycline resistance was most commonly observed in the wild bird, water and zoo isolates. No or negligible resistance to erythromycin or gentamicin was detected. In all data, 12/26 of the tetracycline-resistant isolates were also resistant to streptomycin (P < 0.001) and the clonal complex (CC) ST-1034 CC showed a high proportion of 75% (9/12) of tetracycline-resistant isolates, most originating from the zoo and broilers with closely associated MLST types from these sources. No association between quinolone resistance and MLST type was seen. The low percentage of resistant isolates among the domestic Campylobacter infections is most probably due to the long-term controlled use of antimicrobials. However, the higher percentage of tetracycline resistance observed among the zoo isolates could present a risk for zoo visitors of acquisition of resistant C. jejuni. The resistance pattern of tetracycline and streptomycin most often found in ST-1034 CC could indicate a common resistance acquisition mechanism
Förstner, Konrad U.; Heidrich, Nadja; Reinhardt, Richard; Nieselt, Kay; Sharma, Cynthia M.
Campylobacter jejuni is currently the leading cause of bacterial gastroenteritis in humans. Comparison of multiple Campylobacter strains revealed a high genetic and phenotypic diversity. However, little is known about differences in transcriptome organization, gene expression, and small RNA (sRNA) repertoires. Here we present the first comparative primary transcriptome analysis based on the differential RNA–seq (dRNA–seq) of four C. jejuni isolates. Our approach includes a novel, generic method for the automated annotation of transcriptional start sites (TSS), which allowed us to provide genome-wide promoter maps in the analyzed strains. These global TSS maps are refined through the integration of a SuperGenome approach that allows for a comparative TSS annotation by mapping RNA–seq data of multiple strains into a common coordinate system derived from a whole-genome alignment. Considering the steadily increasing amount of RNA–seq studies, our automated TSS annotation will not only facilitate transcriptome annotation for a wider range of pro- and eukaryotes but can also be adapted for the analysis among different growth or stress conditions. Our comparative dRNA–seq analysis revealed conservation of most TSS, but also single-nucleotide-polymorphisms (SNP) in promoter regions, which lead to strain-specific transcriptional output. Furthermore, we identified strain-specific sRNA repertoires that could contribute to differential gene regulation among strains. In addition, we identified a novel minimal CRISPR-system in Campylobacter of the type-II CRISPR subtype, which relies on the host factor RNase III and a trans-encoded sRNA for maturation of crRNAs. This minimal system of Campylobacter, which seems active in only some strains, employs a unique maturation pathway, since the crRNAs are transcribed from individual promoters in the upstream repeats and thereby minimize the requirements for the maturation machinery. Overall, our study provides new insights into
Jokinen, Cassandra C; Schreier, Hans; Mauro, William; Taboada, Eduardo; Isaac-Renton, Judith L; Topp, Edward; Edge, Thomas; Thomas, James E; Gannon, Victor P J
In this study, we wished to assess the prevalence and determine the sources of three zoonotic bacterial pathogens (Salmonella, Campylobacter, and Escherichia coli O157:H7) in the Salmon River watershed in southwestern British Columbia. Surface water, sewage, and animal faecal samples were collected from the watershed. Selective bacterial culture and PCR techniques were used to isolate these three pathogens and indicator bacteria from these samples and characterize them. Campylobacter was the most prevalent pathogen in all samples, followed by Salmonella, and E. coli O157:H7. E. coli O157:H7 and Salmonella isolation rates from water, as well as faecal coliform densities correlated positively with precipitation, while Campylobacter isolation rates correlated negatively with precipitation. Analysis of DNA extracted from water samples for the presence of Bacteroides host-species markers, and comparisons of C. jejuni flaA-RFLP types and Salmonella serovars from faecal and water samples provided evidence that human sewage and specific domestic and wild animal species were sources of these pathogens; however, in most cases the source could not be determined or more than one source was possible. The frequent isolation of these zoonotic pathogens in the Salmon River highlights the risks to human health associated with intentional and unintentional consumption of untreated surface waters.
González-Hein, Gisela; Huaracán, Bernardo; García, Patricia; Figueroa, Guillermo
Campylobacter jejuni isolates of different origins (bovine, broiler meat, human) were screened by polymerase chain reaction for the presence of 4 genes cdtB, cst-II, ggt, and virB11, previously linked to virulence such as adherence, invasion, colonization, molecular mimicry, and cytotoxin production. In addition, the isolates were screened for the presence of the global gene regulator csrA linked to oxidative stress responses, biofilms formation, and cell adhesion. All the C. jejuni isolates were positive for cdtB gene. The csrA gene was detected in 100% and 92% of C. jejuni isolates from human and animal origin and the virB11 gene was detected in 7.3% and 3.6% isolates from chicken and human respectively. All isolates from bovine were negative for the virB11 gene. The isolates showed a wide variation for the presence of the remaining genes. Of the C. jejuni recovered from human 83.6%, and 32.7% were positive for cst-II, and ggt respectively. Out of the isolates from chicken 40% and 5.5% isolates revealed the presence of cst-II, and ggt, respectively. Finally of the C. jejuni isolates from bovine, 97.7% and 22.7% were positive for cst-II, and ggt respectively. We conclude that the genes of this study circulate among humans and animals. These results led us to hypothesize that the isolates associated with enteritis (cdtB positives) are not selected by environmental or host-specific factors. On the other hand, the high frequencies of csrA gene in C. jejuni show that this gene is important for the survival of C. jejuni in animals and humans.
González-Hein, Gisela; Huaracán, Bernardo; García, Patricia; Figueroa, Guillermo
Campylobacter jejuni isolates of different origins (bovine, broiler meat, human) were screened by polymerase chain reaction for the presence of 4 genes cdtB, cst-II, ggt, and virB11, previously linked to virulence such as adherence, invasion, colonization, molecular mimicry, and cytotoxin production. In addition, the isolates were screened for the presence of the global gene regulator csrA linked to oxidative stress responses, biofilms formation, and cell adhesion. All the C. jejuni isolates were positive for cdtB gene. The csrA gene was detected in 100% and 92% of C. jejuni isolates from human and animal origin and the virB11 gene was detected in 7.3% and 3.6% isolates from chicken and human respectively. All isolates from bovine were negative for the virB11 gene. The isolates showed a wide variation for the presence of the remaining genes. Of the C. jejuni recovered from human 83.6%, and 32.7% were positive for cst-II, and ggt respectively. Out of the isolates from chicken 40% and 5.5% isolates revealed the presence of cst-II, and ggt, respectively. Finally of the C. jejuni isolates from bovine, 97.7% and 22.7% were positive for cst-II, and ggt respectively. We conclude that the genes of this study circulate among humans and animals. These results led us to hypothesize that the isolates associated with enteritis (cdtB positives) are not selected by environmental or host-specific factors. On the other hand, the high frequencies of csrA gene in C. jejuni show that this gene is important for the survival of C. jejuni in animals and humans. PMID:24688515
Pintar, Katarina D. M.; Christidis, Tanya; Thomas, M. Kate; Anderson, Maureen; Nesbitt, Andrea; Keithlin, Jessica; Marshall, Barbara; Pollari, Frank
Canada and abroad. Within this literature, knowledge gaps were identified, and include: a lack of concentration data reported in the literature for Campylobacter spp. in animal feces, a distinction between ill and diarrheic pets in the reported studies, noted differences in shedding and concentrations for various subtypes of Campylobacter, and consistent reporting between studies. PMID:26683667
Pintar, Katarina D M; Christidis, Tanya; Thomas, M Kate; Anderson, Maureen; Nesbitt, Andrea; Keithlin, Jessica; Marshall, Barbara; Pollari, Frank
Canada and abroad. Within this literature, knowledge gaps were identified, and include: a lack of concentration data reported in the literature for Campylobacter spp. in animal feces, a distinction between ill and diarrheic pets in the reported studies, noted differences in shedding and concentrations for various subtypes of Campylobacter, and consistent reporting between studies.
Pham, Ngan Thi Kim; Thongprachum, Aksara; Tran, Dinh Nguyen; Nishimura, Shuichi; Shimizu-Onda, Yuko; Trinh, Quang Duy; Khamrin, Pattara; Ukarapol, Nuthapong; Kongsricharoern, Tipachan; Komine-Aizawa, Shihoko; Okitsu, Shoko; Maneekarn, Niwat; Hayakawa, Satoshi; Ushijima, Hiroshi
A total of 29 Campylobacter jejuni and C. coli strains were isolated from Thai and Japanese children with diarrhea using the Loop-mediated Isothermal Amplification method. The samples were evaluated for mutations in gyrA and 23S rRNA in order to assess resistance against fluoroquinolones and macrolides, respectively. Among the isolated strains, 9 (8 C. jejuni and 1 C. coli) were from Thai children, and the other 20 (C. jejuni) were isolated from Japanese children. High fluoroquinolone resistance rates were observed in Thai (66.7%) and Japanese (90%) children. Macrolide resistance was not observed in Japanese children but was observed at a considerable rate of 12.5% of C. jejuni isolated in the Thai cohort. The results indicate that continuous monitoring of resistance of Campylobacter strains to fluoroquinolones and macrolides is definitely necessary.
Soerjadi-Liem, A S; Snoeyenbos, G H; Weinack, O M
Resistance of young chicks to Campylobacter fetus subsp. jejuni was substantially increased by early exposure to native gut microflora. Protection was demonstrated against two human isolates and a chicken isolate of C. fetus subsp. jejuni. Significant protection against the chicken isolate was observed throughout a 91-day test period. Infection reached 100% (25/25) in the untreated group at 56 days of age and only 4% (1/25) in the group treated with native gut microflora. Campylobacter fetus subsp. jejuni was isolated from the ceca and less frequently from the gall bladder and liver of chicks that actively shed the bacteria. Cultures of feces from chicks reared on wood-shavings litter were often negative, suggesting that culturing litter as an indicator of infection has limited value.
González, L M; Villalobos, N; Montero, E; Morales, J; Sanz, R Alamo; Muro, A; Harrison, L J S; Parkhouse, R M E; Gárate, T
In the present work, the species-specific identification of Taeniid spp. cysticerci and sarcocystis cysts isolated from infected pigs and cattle was achieved by PCR. In particular: (i) multiplex-PCR derived from HDP2 DNA fragment, specific for Taenia saginata/Taenia solium; (ii) PCRs and PCR-RFLPs of the rDNA internal transcribed spacers 1 and 2 (ITS1 and ITS2) for the differential diagnosis of taeniids; (iii) PCR derived from the 18S rRNA gene and sequencing, specific for Sarcoystis spp. The combined application of these three PCR protocols provided an unequivocally specific diagnosis of T. saginata, T. solium, T. hydatigena, Sarcocystis hominis and Sarcocystis suihominis, and may have practical application in the identification of calcified degenerating or morphologically dubious cysts, for example in the slaughter house situation or in human biopsy samples.
Plaza-Rodríguez, C; Appel, B; Kaesbohrer, A; Filter, M
Within the European activities for the 'Monitoring and Collection of Information on Zoonoses', annually EFSA publishes a European report, including information related to the prevalence of Campylobacter spp. in Germany. Spatial epidemiology becomes here a fundamental tool for the generation of these reports, including the representation of prevalence as an essential element. Until now, choropleth maps are the default visualization technique applied in epidemiological monitoring and surveillance reports made by EFSA and German authorities. However, due to its limitations, it seems to be reasonable to explore alternative chart type. Four maps including choropleth, cartogram, graduated symbols and dot-density maps were created to visualize real-world sample data on the prevalence of Campylobacter spp. in raw chicken meat samples in Germany in 2011. In addition, adjacent and coincident maps were created to visualize also the associated uncertainty. As an outcome, we found that there is not a single data visualization technique that encompasses all the necessary features to visualize prevalence data alone or prevalence data together with their associated uncertainty. All the visualization techniques contemplated in this study demonstrated to have both advantages and disadvantages. To determine which visualization technique should be used for future reports, we recommend to create a dialogue between end-users and epidemiologists on the basis of sample data and charts. The final decision should also consider the knowledge and experience of end-users as well as the specific objective to be achieved with the charts.
Hormeño, Lorena; Palomo, Gonzalo; Ugarte-Ruiz, María; Porrero, M Concepción; Borge, Carmen; Vadillo, Santiago; Píriz, Segundo; Domínguez, Lucas; Campos, Maria J; Quesada, Alberto
Among zoonotic diseases, campylobacteriosis stands out as the major bacterial infection producing human gastroenteritis. Antimicrobial therapy, only recommended in critical cases, is challenged by resistance mechanisms that should be unambiguously detected for achievement of effective treatments. Quinolone (ciprofloxacin) resistance of Campylobacter jejuni and Campylobacter coli, the 2 main Campylobacter detected in humans, is conferred by the mutation gyrA C-257-T, which can be genotyped by several methods that require a previous identification of the pathogen species to circumvent the sequence polymorphism of the gene. A multiplex PCR, based on degenerated oligonucleotides, has been designed for unambiguous identification of the quinolone resistance determinant in Campylobacter spp. isolates. The method was verified with 249 Campylobacter strains isolated from humans (141 isolates) and from the 3 most important animal sources for this zoonosis: poultry (34 isolates), swine (38 isolates), and cattle (36 isolates). High resistance to ciprofloxacin, MIC above 4μg/mL, linked to the mutated genotype predicted by MAMA-DEG PCR (mismatch amplification mutation assay PCR with degenerated primers) was found frequently among isolates from the different hosts.
Cha, Wonhee; Mosci, Rebekah; Wengert, Samantha L.; Singh, Pallavi; Newton, Duane W.; Salimnia, Hossein; Lephart, Paul; Khalife, Walid; Mansfield, Linda S.; Rudrik, James T.; Manning, Shannon D.
Campylobacter jejuni is a zoonotic pathogen and the most common bacterial cause of human gastroenteritis worldwide. With the increase of antibiotic resistance to fluoroquinolones and macrolides, the drugs of choice for treatment, C. jejuni was recently classified as a serious antimicrobial resistant threat. Here, we characterized 94 C. jejuni isolates collected from patients at four Michigan hospitals in 2011 and 2012 to determine the frequency of resistance and association with phylogenetic lineages. The prevalence of resistance to fluoroquinolones (19.1%) and macrolides (2.1%) in this subset of C. jejuni isolates from Michigan was similar to national reports. High frequencies of fluoroquinolone-resistant C. jejuni isolates, however, were recovered from patients with a history of foreign travel. A high proportion of these resistant isolates were classified as multilocus sequence type (ST)-464, a fluoroquinolone-resistant lineage that recently emerged in Europe. A significantly higher prevalence of tetracycline-resistant C. jejuni was also found in Michigan and resistant isolates were more likely to represent ST-982, which has been previously recovered from ruminants and the environment in the U.S. Notably, patients with tetracycline-resistant C. jejuni infections were more likely to have contact with cattle. These outcomes prompt the need to monitor the dissemination and diversification of imported fluoroquinolone-resistant C. jejuni strains and to investigate the molecular epidemiology of C. jejuni recovered from cattle and farm environments to guide mitigation strategies. PMID:27199922
Ugarte-Ruiz, M; Stabler, R A; Domínguez, L; Porrero, M C; Wren, B W; Dorrell, N; Gundogdu, O
Infections from Campylobacter jejuni pose a serious public health problem and are now considered the leading cause of foodborne bacterial gastroenteritis throughout the world. Sequencing of C. jejuni genomes has previously allowed a number of loci to be identified, which encode virulence factors that aid survival and pathogenicity. Recently, a Type VI secretion system (T6SS) consisting of 13 conserved genes was described in C. jejuni strains and recognised to promote pathogenicity and adaptation to the environment. In this study, we determined the presence of this T6SS in 63 Spanish C. jejuni isolates from the food chain and urban effluents using whole-genome sequencing. Our findings demonstrated that nine (14%) strains harboured the 13 ORFs found in prototype strain C. jejuni 108. Further studies will be necessary to determine the prevalence and importance of T6SS-positive C. jejuni strains.
Sørensen, Martine C Holst; Gencay, Yilmaz Emre; Birk, Tina; Baldvinsson, Signe Berg; Jäckel, Claudia; Hammerl, Jens A; Vegge, Christina S; Neve, Horst; Brøndsted, Lone
In this study we isolated novel bacteriophages, infecting the zoonotic bacterium Campylobacter jejuni. These phages may be used in phage therapy of C. jejuni colonized poultry to prevent spreading of the bacteria to meat products causing disease in humans. Many C. jejuni phages have been isolated using NCTC12662 as the indicator strain, which may have biased the selection of phages. A large group of C. jejuni phages rely on the highly diverse capsular polysaccharide (CPS) for infection and recent work identified the O-methyl phosphoramidate modification (MeOPN) of CPS as a phage receptor. We therefore chose seven C. jejuni strains each expressing different CPS structures as indicator strains in a large screening for phages in samples collected from free-range poultry farms. Forty-three phages were isolated using C. jejuni NCTC12658, NCTC12662 and RM1221 as host strains and 20 distinct phages were identified based on host range analysis and genome restriction profiles. Most phages were isolated using C. jejuni strains NCTC12662 and RM1221 and interestingly phage genome size (140 kb vs. 190 kb), host range and morphological appearance correlated with the isolation strain. Thus, according to C. jejuni phage grouping, NCTC12662 and NCTC12658 selected for CP81-type phages, while RM1221 selected for CP220-type phages. Furthermore, using acapsular ∆kpsM mutants we demonstrated that phages isolated on NCTC12658 and NCTC12662 were dependent on the capsule for infection. In contrast, CP220-type phages isolated on RM1221 were unable to infect non-motile ∆motA mutants, hence requiring motility for successful infection. Hence, the primary phage isolation strain determines both phage type (CP81 or CP220) as well as receptors (CPS or flagella) recognised by the isolated phages.
Mąka, Łukasz; Popowska, Magdalena
This review summarizes current data on resistance among Salmonella spp. isolates of food origin from countries in different regions of the world. The mechanisms of resistance to different groups of antimicrobial compounds are also considered. Among strains resistant to quinolones and/or fluoroquinolones the most prevalent mechanism is amino acid substitutions in quinolone resistance-determining region (QRDR) of genes gyrA, parC but mechanism of growing importance is plasmid-mediated quinolone resistance (PMQR) associated with genes qnrA, qnrB, qnrC, qnrD, qnrS but frequency of their detection is different. Resistance to sulfonamides is mostly associated with genes sul1 and sul2, while resistance to trimethoprim is associated with various variants of dhfr ( dfr) genes. Taking into account Salmonella spp. strains isolated from food, resistance to β-lactams is commonly associated with β-lactamases encoding by blaTEM genes. However strains ESBL and AmpC – positive are also detected. Resistance to aminoglicosides is commonly result of enzymatic inactivation. Three types of aminoglycoside modifying enzyme are: acetyltransferases (AAC), adenyltransferases (ANT) and phosphotransferases (APH). Resistance to tetracyclines among Salmonella spp. isolated from food is most commonly associated with active efflux. Among numerous genetic determinants encoding efflux pumps tetA, tetB, tetC, tetD, tetE and tetG are reported predominatingly. One of the most common mechanisms of resistance against chloramphenicol is its inactivation by chloramphenicol acetyltrasferases (CATs), but resistance to this compound can be also mediated by chloramphenicol efflux pumps encoded by the genes cmlA and floR. It is important to monitor resistance of Salmonella isolated from food, because the globalization of trade, leading to the long-distance
We have purified a bacteriocin peptide (termed CUV-3), produced by a poultry cecal isolate of Campylobacter jejuni (strain CUV-3) with inhibitory activity against Gram positive bacteria including Clostridium perfringens (38 strains), Staphylococcus aureus, Staphylococcus epidermidis and Listeria mon...
Sippy, Rachel; Sandoval-Green, Claudette M J; Sahin, Orhan; Plummer, Paul; Fairbanks, W Sue; Zhang, Qijing; Blanchong, Julie A
Wildlife harbor a variety of Campylobacter spp. and may play a significant role in the transmission of Campylobacter to livestock. Although studies have been conducted on wildlife-associated Campylobacter isolates from farms in other countries, there are little data available for livestock farms in the United States. In addition, the critical questions of whether wildlife harbor Campylobacter that is pathogenic to ruminants and/or antibiotic-resistant Campylobacter have yet to be addressed. We captured wild small mammals (n=142) and small birds (n=188) at livestock farms in central Iowa and sampled them for thermophilic Campylobacter during autumn 2009, spring 2010, and autumn 2010. Overall prevalence was 4.79%, with isolates found only in wild birds. Molecular typing revealed four multilocus sequence types (STs), three of which are novel. The remaining ST (ST-806) was found in two house sparrows and is an ST previously associated with ruminant abortion cases. Further analysis of ST-806 wild bird and ruminant abortion isolates by pulsed-field gel electrophoresis, resistance gene location, and antibiotic susceptibility tests indicated that the isolates are nearly identical. This is the first account of isolation of Campylobacter types from wild birds that are known to be pathogenic to ruminants. Furthermore, these same two wild bird isolates are resistant to the antibiotic fluoroquinolone. Our results indicate there is an overall low prevalence of Campylobacter in selected wildlife in Iowa, but suggest that wildlife may play a role in the epidemiology of pathogenic Campylobacter for domestic livestock, and may also serve as a reservoir for antibiotic-resistant Campylobacter.
Background Campylobacter concisus is an emerging enteric pathogen, yet it is commonly isolated from feces and the oral cavities of healthy individuals. This genetically complex species is comprised of several distinct genomospecies which may vary in pathogenic potential. Results We compared pathogenic and genotypic properties of C. concisus fecal isolates from diarrheic and healthy humans residing in the same geographic region. Analysis of amplified fragment length polymorphism (AFLP) profiles delineated two main clusters. Isolates assigned to AFLP cluster 1 belonged to genomospecies A (based on genomospecies-specific differences in the 23S rRNA gene) and were predominantly isolated from healthy individuals. This cluster also contained a reference oral strain. Isolates assigned to this cluster induced greater expression of epithelial IL-8 mRNA and more frequently contained genes coding for the zonnula occludins toxin and the S-layer RTX. Furthermore, isolates from healthy individuals induced greater apoptotic DNA fragmentation and increased metabolic activity than those from diarrheic individuals, and isolates assigned to genomospecies A (of which the majority were from healthy individuals) exhibited higher haemolytic activity compared to genomospecies B isolates. In contrast, AFLP cluster 2 was predominated by isolates belonging to genomospecies B and those from diarrheic individuals. Isolates from this cluster displayed greater mean epithelial invasion and translocation than cluster 1 isolates. Conclusion Two main genetically distinct clusters (i.e., genomospecies) were identified among C. concisus fecal isolates from healthy and diarrheic individuals. Strains within these clusters differed with respect to clinical presentation and pathogenic properties, supporting the hypothesis that pathogenic potential varies between genomospecies. ALFP cluster 2 isolates were predominantly from diarrheic patients, and exhibited higher levels of epithelial invasion and
The transmission of Campylobacter spp. and baseline level of antimicrobial resistance associated with these organisms has significant implications for environmental, animal, and human health. One focus is the use of antibiotics in animal agriculture and the effects on antibiotic resistant bacterial ...
Klein-Jöbstl, Daniela; Sofka, Dmitri; Iwersen, Michael; Drillich, Marc; Hilbert, Friederike
Human campylobacteriosis is primarily associated with poultry but also cattle. In this study, 55 Campylobacter jejuni strains isolated from 382 dairy calves' feces were differentiated by multilocus sequence typing and tested for antimicrobial resistance. The most prevalent sequence type (ST) was ST883 (20.0%), followed by ST48 (14.5%), and ST50 (9.1%). In contrast to ST48 and ST50, ST883 has rarely been described in cattle previously. Furthermore, risk factor analysis was performed for the presence of the most prevalent STs in these calves. Multiple regression analysis revealed that the type of farm (organic vs. conventional) and calf housing (place, and individual vs. group) were identified as significantly (p < 0.05) associated with the presence of ST883 in calves, and ST50 was associated with calf diarrhea. Antimicrobial resistance was detected in 58.2% of the isolates. Most of the resistant isolates (81.3%) were resistant to more than one antimicrobial. Most frequently, resistance to ciprofloxacin (49.1%), followed by nalidixic acid (42.8%), and tetracycline (14.5%) was observed. The results of the present study support the hypothesis that dairy calves may serve as a potential reservoir for C. jejuni and pose a risk for transmission, including antimicrobial resistant isolates to the environment and to humans.
Klein-Jöbstl, Daniela; Sofka, Dmitri; Iwersen, Michael; Drillich, Marc; Hilbert, Friederike
Human campylobacteriosis is primarily associated with poultry but also cattle. In this study, 55 Campylobacter jejuni strains isolated from 382 dairy calves’ feces were differentiated by multilocus sequence typing and tested for antimicrobial resistance. The most prevalent sequence type (ST) was ST883 (20.0%), followed by ST48 (14.5%), and ST50 (9.1%). In contrast to ST48 and ST50, ST883 has rarely been described in cattle previously. Furthermore, risk factor analysis was performed for the presence of the most prevalent STs in these calves. Multiple regression analysis revealed that the type of farm (organic vs. conventional) and calf housing (place, and individual vs. group) were identified as significantly (p < 0.05) associated with the presence of ST883 in calves, and ST50 was associated with calf diarrhea. Antimicrobial resistance was detected in 58.2% of the isolates. Most of the resistant isolates (81.3%) were resistant to more than one antimicrobial. Most frequently, resistance to ciprofloxacin (49.1%), followed by nalidixic acid (42.8%), and tetracycline (14.5%) was observed. The results of the present study support the hypothesis that dairy calves may serve as a potential reservoir for C. jejuni and pose a risk for transmission, including antimicrobial resistant isolates to the environment and to humans. PMID:26870027
Szczepańska, Bernadeta; Kamiński, Piotr; Andrzejewska, Małgorzata; Śpica, Dorota; Kartanas, Edmund; Ulrich, Werner; Jerzak, Leszek; Kasprzak, Mariusz; Bocheński, Marcin; Klawe, Jacek J
The aim of this study was to investigate the role of white stork Ciconia ciconia as a potential reservoir of Campylobacter spp. Antimicrobial resistance and the presence of putative virulence genes of the isolates were also examined. A total of 398 white stork chicks sampled in Western Poland in habitats with high density of breeding were examined. Rectal swabs were collected during breeding season 2009-2012 from storks developing in a relatively pure environment (Odra meadows), in polluted areas (a copper mining-smelting complex), and in suburbs. Of the anal swabs collected, 7.6% were positive for Campylobacter among chicks (5.3% samples positive for C. jejuni and 2.3% samples positive for C. coli). Samples from polluted areas had the highest prevalence of Campylobacter (12.2%). The prevalence of resistance among C. jejuni and C. coli isolates from young storks was as follows: to ciprofloxacin (52.4%, 44.4%), and to tetracycline (19%, 77.8%). All of the analyzed isolates were susceptible to macrolides. The resistance to both classes of antibiotics was found in the 23.3% of Campylobacter spp. All Campylobacter spp. isolates had cadF gene and flaA gene responsible for adherence and motility. CdtB gene associated with toxin production was present in 88.9% of C. coli isolates and 57.1% of C. jejuni isolates. The iam marker was found more often in C. coli strains (55.6%) compared to C. jejuni isolates (42.9%). Our results confirm the prevalence of Campylobacter spp. in the white stork in natural conditions and, because it lives in open farmlands with access to marshy wetlands, the environmental sources such as water reservoirs and soil-water can be contaminated from white stork feces and the pathogens can be widely disseminated. We can thus conclude that Campylobacter spp. may easily be transmitted to waterfowl, other birds, and humans via its environmental sources and/or by immediate contact.
Nawaz, Mohamed; Khan, S A; Tran, Q; Sung, K; Khan, A A; Adamu, I; Steele, R S
A study was undertaken to isolate and characterize tetracycline and nalidixic acid-resistant Klebsiella spp. in farm-raised, imported shrimp sold in the United States. Sixty-seven multiple antibiotic-resistant Klebsiella spp. strains were isolated from imported shrimp samples. Using morphological and biochemical methods, fifty-three strains were tentatively identified as Klebsiella pneumoniae and fourteen as K. oxytoca. Although all isolates were resistant to tetracycline, only 8 were resistant to nalidixic acid. These 8 isolates were further screened by PCR for quinolone resistance genes (qnrA, B, S, gyrA, B and parC). PCR protocols failed to amplify any qnr genes. The purified PCR amplicons of gyrA, gyrB and parC were sequenced and analyzed for point mutations that confer resistance to fluoroquinolone antibiotics. Analysis of the sequences of the gyrA amplicons from nalidixic acid-resistant Klebsiella spp. indicated two point mutations in gyrA at positions 83 (Ser→Phe) and 87 (Asp→Ala). Sequence analysis of the parC amplicons indicated an amino acid change at position 80 (Ser→Ile). No mutations were detected in gyrB. Template DNA from all isolates was screened for tetracycline resistance genes (tetA-E). Oligonucleotide primers specifically targeting a 305-bp region of tetB and a 477-bp region of tetD successfully amplified sequences from 91.0 and 44.0% of the isolates, respectively. None of the isolates contained tetA, tetC or tetE genes. Plasmids (2.0-16.0kb) were found in 23 of the 67 isolates. XbaI-PFGE identified 32 distinct macro restriction patterns (mrps) among the 61 multiple drug-resistant Klebsiella spp. that were typable. Our results indicate that imported shrimp is a reservoir for multidrug resistant Klebsiella spp. and potential health risks posed by such strains should not be underestimated.
Sandalakis, Vassilios; Chochlakis, Dimosthenis; Goniotakis, Ioannis; Tselentis, Yannis; Psaroulaki, Anna
In Greece standard tests are performed in the watering and cooling systems of hotels' units either as part of the surveillance scheme or following human infection. The purpose of this study was to establish the minimum inhibitory concentration (MIC) distributions of environmental Legionella isolates for six antimicrobials commonly used for the treatment of Legionella infections, by MIC-test methodology. Water samples were collected from 2004 to 2011 from 124 hotels from the four prefectures of Crete (Greece). Sixty-eight (68) Legionella isolates, comprising L. pneumophila serogroups 1, 2, 3, 5, 6, 8, 12, 13, 15, L. anisa, L. rubrilucens, L. maceachernii, L. quinlivanii, L. oakridgensis, and L. taurinensis, were included in the study. MIC-tests were performed on buffered charcoal yeast extract with α-ketoglutarate, L-cysteine, and ferric pyrophosphate. The MICs were read after 2 days of incubation at 36 ± 1 °C at 2.5% CO2. A large distribution in MICs was recorded for each species and each antibiotic tested. Rifampicin proved to be the most potent antibiotic regardless of the Legionella spp.; tetracycline appeared to have the least activity on our environmental isolates. The MIC-test approach is an easy, although not so cost-effective, way to determine MICs in Legionella spp. These data should be kept in mind especially since these Legionella species may cause human disease.
Background The Cryptococcus spp is currently composed of encapsulated yeasts of cosmopolitan distribution, including the etiological agents of cryptococcosis. The fungus are found mainly in substrates of animal and plant origin. Human infection occurs through inhalation of spores present in the environment. Methods Eighty-four swab collections were performed on dust found on books in three libraries in the city of Cuiabá, state of Mato Grosso, Brazil. The material was seeded in Sabouraud agar and then observed for characteristics compatible with colonies with a creamy to mucous aspect; the material was then isolated in birdseed (Niger) agar and cultivated at a temperature of 37°C for 5 to 7 days. Identification of isolated colonies was performed by microscopic observation in fresh preparations dyed with India ink, additional tests performed on CGB (L-canavanine glycine bromothymol blue), urea broth, and carbohydrate assimilation tests (auxanogram). Results Of the 84 samples collected from book dust, 18 (21.4%) were positive for Cryptococcus spp totalizing 41 UFC’s. The most frequently isolated species was C. gattii 15 (36.6%); followed by C. terreus, 12 (29.3%); C. luteolus 4 (9.8%); C. neoformans, and C. uniguttulatus 3 (7.3%), and C. albidus and C. humiculus with 2 (4.6%) of the isolates. Conclusion The high biodiversity of the yeasts of the Cryptococcus genus, isolated from different environmental sources in urban areas of Brazil suggests the possibility of individuals whose immune systems have been compromised or even healthy individuals coming into sources of fungal propagules on a daily bases throughout their lives. This study demonstrates the acquisition possible of cryptococcosis infection from dust in libraries. PMID:22682392
Abay, Secil; Kayman, Tuba; Otlu, Baris; Hizlisoy, Harun; Aydin, Fuat; Ertas, Nurhan
In this study, the investigation of clonal relations between human and poultry Campylobacter jejuni isolates and the determination of susceptibilities of isolates to various antibiotics were aimed. A total of 200 C. jejuni isolates concurrently obtained from 100 chicken carcasses and 100 humans were genotyped by the Pulsed-Field Gel Electrophoresis (PFGE) and automated Repetitive Extragenic Palindromic PCR (Rep-PCR, DiversiLab system) methods and were tested for their susceptibility to six antibiotics with disk diffusion method. The minimum inhibitory concentration (MIC) values of ciprofloxacin (CI), enrofloxacin (EF) and erythromycin (EM) were evaluated by E-test. By using PFGE 174 of (87.0%) the isolates were able to be typed. The clonally related strains were placed in 35 different clusters and 115 different genotypes were obtained. All of the two hundred isolates could be typed by using Rep-PCR and were divided into 133 different genotypes. One hundred and fourteen clonally related isolates (57.0%) were included in 47 clusters. In disk diffusion test, while the susceptibility rates of AMC and S to human and chicken derived C. jejuni isolates were 84.0%-96.0% and 96.0%-98.0%, respectively, all isolates were susceptible to gentamicin. The resistance rates of human isolates to AMP, NA and TE were detected as 44.0%, 84.0% and 38.0% of the resistances of chicken isolates to these antibiotics were 34.0%, 95.0% and 56.0%, respectively. The MIC values of human and chicken isolates to CI, EF and EM were detected as 81.0-93.0%, 85.0-88.0% and 6.0-7.0%, respectively. The clonal proximity rates were detected between human and poultry origin C. jejuni isolates. The discriminatory power of PFGE and Rep-PCR was similar, with Simpson's diversity indexes of 0.993 and 0.995, respectively. Concordance of the two methods as determined by Adjusted Rand coefficient was 0.198 which showed the low congruence between Rep-PCR and PFGE. High rates of quinolone resistance were detected in
Shin, Eunju; Oh, Younghee; Kim, Moosang; Jung, Jihun; Lee, Yeonhee
A total of 121 Campylobacter isolates from 4,788 humans with gastroenteritis were identified and characterized by biochemical detection methods, polymerase chain reaction, and multilocus sequence typing (MLST). These samples were obtained during a 3-year period, from January 2007 to December 2009, using the National Notifiable Diseases Surveillance System at the Research Institute of Public Health and Environment in Seoul Metropolitan, Korea. Antimicrobial susceptibilities of the bacterium were also determined with the agar dilution method. All 121 isolates were identified as Campylobacter jejuni, with all (100%) of them having two virulence genes (ceuE and cadF) and a toxin gene (cdtB). Twenty-three different sequence types (STs), including 9 new STs, were determined by MLST. The most prevalent ST and clonal complex (CC) observed in this study were ST-45 (28.9%) and ST-45 CC (53.7%), respectively. Percentages of antimicrobial-resistant isolates were 1.9% for ampicillin, 0.8% for chloramphenicol, 24% for ciprofloxacin, 46.3% for enrofloxacin, 0.8% for erythromycin, 6.6% for gentamicin, and 46.3% for tetracycline. This study demonstrated that the majority of the Campylobacter isolates obtained from human samples in Korea were C. jejuni with ST-45 CC, which has been detected mainly in broilers worldwide, and all strains with new STs were uniformly resistant to enrofloxacin and tetracycline. This study indicates that broilers may be a breeding ground for bacteria as well as an important potential source of human campylobacteriosis.
Hao, Haihong; Ren, Ni; Han, Jing; Foley, Steven L.; Iqbal, Zahid; Cheng, Guyue; Kuang, Xiuhua; Liu, Jie; Liu, Zhenli; Dai, Menghong; Wang, Yulian; Yuan, Zonghui
The aim of this study was to reveal the molecular mechanism involved in multidrug resistance and virulence of Campylobacter jejuni isolated from broiler chickens. The virulence of six multidrug resistant C. jejuni was determined by in vitro and in vivo methods. The de novo whole genome sequencing technology and molecular biology methods were used to analyze the genomic features associated with the multidrug resistance and virulence of a selected isolate (C. jejuni 1655). The comparative genomic analyses revealed a large number of single nucleotide polymorphisms, deletions, rearrangements, and inversions in C. jejuni 1655 compared to reference C. jejuni genomes. The co-emergence of Thr-86-Ile mutation in gyrA gene, A2075G mutation in 23S rRNA gene, tetO, aphA and aadE genes and pTet plasmid in C. jejuni 1655 contributed its multidrug resistance to fluoroquinolones, macrolides, tetracycline, and aminoglycosides. The combination of multiple virulence genes may work together to confer the relative higher virulence in C. jejuni 1655. The co-existence of mobile gene elements (e.g., pTet) and CRISPR-Cas system in C. jejuni 1655 may play an important role in the gene transfer and immune defense. The present study provides basic information of phenotypic and genomic features of C. jejuni 1655, a strain recently isolated from a chicken displaying multidrug resistance and relatively high level of virulence. PMID:27790202
Background Campylobacter jejuni strain 11168 was demonstrated to have a broad specificity for eukaryotic surface glycosylation using glycan array analysis. The initial screen indicated that sialic acid and mannose are important binding partners after environmental stress, while galactose and fucose structures are likely to be involved in persistent infection. Results In this broader study, five additional human/clinical isolates and six chicken isolates were fully assessed to determine their glycan binding capacity using an extended glycan array. C. jejuni 11168 was rescreened here due to the presence of glycoaminoglycan (GAG) and other structures that were not available on our previous glycan array. The current array analysis of additional C. jejuni strains confirmed the growth condition dependent differences in glycan binding that was previously observed for C. jejuni 11168. We noted strain to strain variations, particularly for the human isolates C. jejuni 520 and 81116 and the chicken isolate C. jejuni 331, with the majority of differences observed in galactose, mannose and GAG binding. Chicken isolates were found to bind to a broader range of glycans compared to the human isolates, recognising branched mannose and carageenan (red seaweed) glycans. Glycan array data was confirmed using cell-based lectin inhibition assays with the fucose (UEA-I) and mannose (ConA) binding lectins. Conclusions This study confirms that all C. jejuni strains tested bind to a broad range of glycans, with the majority of strains (all except 81116) altering recognition of sialic acid and mannose after environmental stress. Galactose and fucose structures were bound best by all strains when C. jejuni was grown under host like conditions confirming the likelihood of these structures being involved in persistent infection. PMID:24119179
Lim, Patrick Wilson N; Tiam-Lee, Daphne C; Paclibare, Phyllis Anne P; Subejano, Ma Socorro Edden P; Cabero-Palma, Juvy Ann S; Penuliar, Gil M
A total of 265 chicken parts were collected from 15 wet markets and 15 supermarkets in Metro Manila, Philippines. Campylobacter spp. was isolated on modified charcoal cefoperazone deoxycholate agar plates and identified through biochemical tests and PCR amplification of genus and species specific genes. Antimicrobial resistance profiles were determined following Clinical and Laboratory Standards Institute protocols. Two hundred seven (78%) Campylobacter spp. were isolated. C. jejuni and C. coli were detected from 170 (64%) and 32 (12%) of the samples, respectively. Liver and skin samples had the greatest level of contamination. Most of the isolates were resistant to clindamycin (98.6%), erythromycin (98.6%), nalidixic acid (98.1%), tetracycline (94.2%), gentamicin (65.2%), and chloramphenicol (52.6%). The results indicated that poultry meat sold in markets in Metro Manila is contaminated with drug resistant Campylobacter.
Amon, P.; Klein, D.; Springer, B.; Jelovcan, S.; Sofka, D.; Hilbert, F.
Campylobacter jejuni is a major cause of the Guillain–Barré syndrome (GBS) and related diseases. These autoimmune diseases are caused by antibodies cross-reacting with the peripheral (GBS) and central neural tissue (Miller Fisher syndrome – MFS, Bicker-staff’s brainstem encephalitis – BBE), leading to acute polyneuropathy. Recently, specific gene loci in C. jejuni have been distinguished which are associated with the onset of GBS, despite a molecular or phenotypic clustering. In this study, we used PCR to analyse C. jejuni isolates of different origin (i.e. bovine, poultry, human) for these genes. A total of 196 isolates were tested for cst-II and neuA. Of these, 101 isolates harboured the cst-II locus and 102 the neuA locus. Eighty-six isolates (44%) hold both genes. The frequency of cst-II in different sources of isolates of bovine, poultry and human isolates did not vary significantly (52, 50 and 52%, respectively). In contrast, the neuA locus was less often found in poultry isolates. Two human strains – from a family outbreak of campylobacteriosis (in 1989 in Austria) in which one person developed MFS – harboured both genes. Thus, although only one in more than 3000 patients with Campylobacter-associated enteritis develop GBS, about half of Campylobacter jejuni strains found in different environments are possibly able to cause GBS. These strains almost equally distributed in bovine, poultry and human isolates. Our results suggest that isolates associated with GBS are not selected by environmental or host-specific factors. Accordingly, this study indicates that host factors such as humoral and cellular immunity are possibly responsible for the development of these autoimmune diseases. PMID:24611117
Wu, Zuowei; Sippy, Rachel; Sahin, Orhan; Plummer, Paul; Vidal, Ana; Newell, Diane; Zhang, Qijing
Campylobacter infection is a leading cause of ovine abortion worldwide. Historically, genetically diverse Campylobacter fetus and Campylobacter jejuni strains have been implicated in such infections, but since 2003 a highly pathogenic, tetracycline-resistant C. jejuni clone (named SA) has become the predominant cause of sheep abortions in the United States. Whether clone SA was present in earlier U.S. abortion isolates (before 2000) and is associated with sheep abortions outside the United States are unknown. Here, we analyzed 54 C. jejuni isolates collected from U.S. sheep abortions at different time periods and compared them with 42 C. jejuni isolates associated with sheep abortion during 2002 to 2008 in Great Britain, using multilocus sequence typing (MLST), pulsed-field gel electrophoresis (PFGE), and array-based comparative genomic hybridization (CGH). Although clone SA (ST-8) was present in the early U.S. isolates, it was not as tetracycline resistant (19% versus 100%) or predominant (66% versus 91%) as it was in the late U.S isolates. In contrast, C. jejuni isolates from Great Britain were genetically diverse, comprising 19 STs and lacking ST-8. PFGE and CGH analyses of representative strains further confirmed the population structure of the abortion isolates. Notably, the Great Britain isolates were essentially susceptible to most tested antibiotics, including tetracycline, while the late U.S. isolates were universally resistant to this antibiotic, which could be explained by the common use of tetracyclines for control of sheep abortions in the United States but not in Great Britain. These results suggest that the dominance of clone SA in sheep abortions is unique to the United States, and the use of tetracyclines may have facilitated selection of this highly pathogenic clone.
Glupczynski, Y; Delmee, M; Bruck, C; Labbe, M; Avesani, V; Burette, A
Forty-nine isolates of Campylobacter pylori were tested for their susceptibility to twenty antibiotics and four anti-ulcer agents by an agar dilution technique. Penicillin and amoxycillin were the most active drugs (MIC90, 0.06 microgram/ml); erythromycin, cefazolin, minocycline, ciprofloxacin, ofloxacin and gentamicin were slightly less active (MIC90, less than or equal to 1 microgram/ml). Moderate activity was found for doxycyclin, rifampin, nitrofurantoin, norfloxacin, pefloxacin, enoxacin, paromomycin, metronidazole and tinidazole. All strains were resistant to trimethoprim (MIC greater than 512 micrograms/ml). Nalidixic acid (MIC90, greater than 256 micrograms/ml) and colistin (MIC90, greater than 64 micrograms/ml) had little to no activity. Of four anti-ulcer drugs, only bismuth subcitrate showed activity (MIC90, 64 micrograms/ml). Strains resistant to all 4-quinolones were found in patients who had previously received ofloxacin as part of a clinical trial aimed at eradication of C. pylori. These isolates remained susceptible to amoxycillin, tetracyclines and to other classes of antibiotics.
Schmutz, Claudia; Mäusezahl, Daniel; Jost, Marianne; Baumgartner, Andreas; Mäusezahl-Feuz, Mirjam
Clinical isolates of Campylobacter spp. and Salmonella spp. are notifiable in Switzerland. In 1995, Campylobacter replaced Salmonella as the most frequently reported food-borne pathogen. We analysed notification data (1988-2013) for these two bacterial, gastrointestinal pathogens of public health importance in Switzerland. Notification rates were calculated using data for the average resident population. Between 1988 and 2013, notified campylobacteriosis cases doubled from 3,127 to 7,499, while Salmonella case notifications decreased, from 4,291 to 1,267. Case notifications for both pathogens peaked during summer months. Campylobacter infections showed a distinct winter peak, particularly in the 2011/12, 2012/13 and 2013/14 winter seasons. Campylobacter case notifications showed more frequent infection in males than females in all but 20-24 year-olds. Among reported cases, patients' average age increased for campylobacteriosis but not for salmonellosis. The inverse trends observed in case notifications for the two pathogens indicate an increase in campylobacteriosis cases. It appears unlikely that changes in patients' health-seeking or physicians' testing behaviour would affect Campylobacter and Salmonella case notifications differently. The implementation of legal microbiological criteria for foodstuff was likely an effective means of controlling human salmonellosis. Such criteria should be decreed for Campylobacter, creating incentives for producers to lower Campylobacter prevalence in poultry.
Ingresa-Capaccioni, S; Jiménez-Trigos, E; Marco-Jiménez, F; Catalá, P; Vega, S; Marin, C
While horizontal transmission is a route clearly linked to the spread of Campylobacter at the farm level, few studies support the transmission of Campylobacter spp. from breeder flocks to their offspring. Thus, the present study was carried out to investigate the possibility of vertical transmission. Breeders were monitored from the time of housing day-old chicks, then throughout the laying period (0 to 60 wk) and throughout their progeny (broiler fattening, 1 to 42 d) until slaughter. All samples were analyzed according with official method ISO 10272:2006. Results revealed that on breeder farms, Campylobacter isolation started from wk 16 and reached its peak at wk 26, with 57.0% and 93.2% of positive birds, respectively. After this point, the rate of positive birds decreased slightly to 86.0% at 60 wk. However, in broiler production all day-old chicks were found negative for Campylobacter spp, and the bacteria was first isolated at d 14 of age (5.0%), with a significant increase in detection during the fattening period with 62% of Campylobacter positive animals at the end of the production cycle. Moreover, non-positive sample was determined from environmental sources. These results could be explained because Campylobacter may be in a low concentration or in a non-culturable form, as there were several studies that successfully detected Campylobacter DNA, but failed to culture. This form can survive in the environment and infect successive flocks; consequently, further studies are needed to develop more modern, practical, cost-effective and suitable techniques for routine diagnosis.
Lycken, Lena; Borch, Elisabeth
Of 42 spoiled cheese spread products, 35 were found to harbor Clostridium spp. Typical signs of spoilage were gas production and off-odor. The identity was determined for about half of the isolates (n = 124) by Analytab Products (API), Biolog, the RiboPrinter System, 16S rDNA sequencing, cellular fatty acid analysis, or some combination of these. The majority of isolates were identified as Clostridium sporogenes (in 33% of products), but Clostridium cochlearium (in 12% of products) and Clostridium tyrobutyricum (in 2% of products) were also retrieved. Similarity analysis of the riboprint patterns for 21 isolates resulted in the identification of 10 ribogroups. A high degree of relatedness was observed between isolates of C. sporogenes originating from products produced 3 years apart, indicating a common and, over time, persistent source of infection. The spoilage potential of 11 well-characterized isolates and two culture collection strains was analyzed by inoculating shrimp cheese spread with single cultures and then storing them at 37 degrees C. Tubes inoculated with C. tyrobutyricum did not show any visible signs of growth (e.g., coagulation, discoloration, gas formation) in the cheese spread. After 2 weeks of incubation, tubes inoculated with C. cochlearium or C. sporogenes showed gas-holes, syneresis with separation of coagulated casein and liquid, and a change in color of the cheese. The amount of CO2 produced by C. cochlearium strains was approximately one-third that produced by the majority of C. sporogenes strains. To our knowledge, this is the first study to isolate and identify C. cochlearium as a spoilage organism in cheese spread.
Polat, Zubeyda Akin; Karakus, Gulderen
Acanthamoeba keratitis (AK) is a potentially devastating and sight-threatening infection of the cornea caused by the ubiquitous free-living amoebae, Acanthamoeba species. Its eradication is difficult because the amoebas encyst, making it highly resistant to anti-amoebic drugs. Acriflavine neutral (ACF) has been used for treatment of microbial infections for humans and fishes. The aim of our study was to evaluate the time-dependent cytotoxicities of ACF against Acanthamoeba spp. Trophozoites and cysts of three different strains (strain PAT06 Acanthamoeba castellanii, strain 2HH Acanthamoeba hatchetti, and strain 11DS A. hatchetti) of Acanthamoeba spp. were tested. All strains had been isolated from patients suffering from a severe AK. The effects of the ACF with the concentrations ranging from 15 to 500 mg mL(-1) on the cytotoxicity of Acanthamoeba strains were examined. ACF showed a time- and dose-dependent amebicidal action on the trophozoites and cysts. Pat06 (A. castellanii) was the most resistant, while strain 11DS (A. hatchetti) was the most sensitive. As a result, ACF could be concluded as a new agent for the treatment of Acanthamoeba infections. On the other hand, it still needs to be further evaluated by in vivo test systems to confirm the efficiency of its biological effect.
Manfreda, Gerardo; Parisi, Antonio; De Cesare, Alessandra; Mion, Domenico; Piva, Silvia; Zanoni, Renato G
In this retrospective study, typing ability, discriminatory power, and concordance between typing results obtained on 123 Campylobacter jejuni turkey isolates, collected in 1998, within 14 different farms, applying multilocus sequence typing (MLST), pulsed-field gel electrophoresis (PFGE), antibiotic resistance profile, and virulence gene pattern, were assessed and compared. Overall, 33 sequence types, 28 pulsotypes, 10 resistotypes, and 5 pathotypes were identified. MLST and PFGE showed the better discriminatory ability (i.e., Simpson's diversity index >0.90) as well as unidirectional (i.e., Wallace and adjusted Wallace coefficients >0.86) and bidirectional (i.e., adjusted Rand coefficient >0.60) concordance. Moreover, both methods showed a good unidirectional and bidirectional concordance with the resistotype. On the contrary, the congruence of both genotyping methods and resistotype with the pathotype seemed due to chance alone. A clonal relationship was identified among 66.7% of the isolates. Furthermore, 59.7% of the investigated isolates were resistant to two or more antimicrobials and 92% to tetracycline. All the isolates harbored cadF and pldA genes, whereas a flaA gene product and a cdtB gene product were amplified from 85.4% and 79.7% of the isolates, respectively, using the primers designed by Bang et al. (2003). The results of this study clarify the level of genetic diversity among the C. jejuni originating from turkeys. MLST level of correlation with PFGE, resistotype, and pathotype is assessed. This result supports the selection of type and number of typing methods to use in epidemiological studies. Finally, the identification of clonal complexes (i.e., groups of profiles differing by no more than one gene from at least one other profile of the group using the entire Campylobacter MLST database) shared between turkey and human isolates suggests that turkeys could be a possible source of Campylobacter infection.
Janda, W M; Morello, J A; Lerner, S A; Bohnhoff, M
Oropharyngeal, urethral, and rectal cultures for pathogenic Neisseria spp. were collected from 815 homosexual men attending a community clinic in Chicago. Meningococci were characterized by serogrouping and antimicrobial susceptibility testing. Gonococci were auxotyped, and susceptibilities to penicillin and tetracycline were determined. Of the 815 men tested, 42.5% carried meningococci in the oropharynx. Gonococci were recovered from the urethra, rectum, and oropharynx of 18.5, 16.3, and 5.6%, respectively. Meningococci were also recovered from the urethra (6 patients) and the rectum (15 patients). Some of these isolates were identical to the isolates from the oropharynges of the same patients, whereas others were distinct from the oropharyngeal isolates by serogroup or antimicrobial susceptibilities. Serogroups B, W135, and C comprised over 90% of the meningococci. Almost 80% of the gonococcal strains required minimal inhibitory concentrations greater than 0.06 micrograms of penicillin per ml, whereas greater than 90% of the meningococci were inhibited at this concentration. Auxotyping demonstrated three major auxotypes: Zero (required none of the nutrients tested), 60%; arginine requiring, 19.4%; and proline requiring, 12.3%. Only four strains (1.2%) required arginine, hypoxanthine, and uracil. PMID:6826712
Kim, Jung-Beom; Kang, Suk-Ho; Park, Yong-Bae; Choi, Jae-Ho; Park, Sung-Jin; Cho, Seung-Hak; Park, Mi-Sun; Lee, Hae Kyung; Choi, Na-Jung; Kim, Ha-Na; Oh, Deog-Hwan
This study was conducted to investigate the phenotypic and genotypic characteristics of Korean isolates of Cronobacter spp. (Enterobacter sakazakii). A total of 43 Cronobacter spp., including 5 clinical isolates, 34 food isolates, 2 environmental isolates, and 2 reference strains (C. sakazakii ATCC 29004 and C. muytjensii ATCC51329) were used in this study. Korean isolates of Cronobacter spp. were divided into 11 biogroups according to their biochemical profiles and 3 genomic groups based on the analysis of their 16S rRNA gene sequences. Biogroups 1 and 2 contained the majority of isolates (n=26), most of which were contained in 16S rRNA cluster 1 (n=34). Korean isolates of Cronobacter spp. showed diverse biochemical profiles. Biogroup 1 contained C. sakazakii GIHE (Gyeonggido Research Institute of Health and Environment) 1 and 2, which were isolated from babies that exhibited symptoms of Cronobacter spp. infection such as gastroenteritis, sepsis, and meningitis. Our finding revealed that Biogroup 1, C. sakazakii, is more prevalent and may be a more pathogenic biogroup than other biogroups, but the pathogenic biogroup was not represented clearly among the 11 biogroups tested in this study. Thus, all biogroups of Cronobacter spp. were recognized as pathogenic bacteria, and the absence of Cronobacter spp. in infant foods should be constantly regulated to prevent food poisoning and infection caused by Cronobacter spp.
Müllner, Petra; Collins-Emerson, Julie M; Midwinter, Anne C; Carter, Philip; Spencer, Simon E F; van der Logt, Peter; Hathaway, Steve; French, Nigel P
In New Zealand the number of campylobacteriosis notifications increased markedly between 2000 and 2007. Notably, this country's poultry supply is different than that of many developed countries as the fresh and frozen poultry available at retail are exclusively of domestic origin. To examine the possible link between human cases and poultry, a sentinel surveillance site was established to study the molecular epidemiology of Campylobacter jejuni over a 3-year period from 2005 to 2008 using multilocus sequence typing. Studies showed that 60.1 to 81.4% of retail poultry carcasses from the major suppliers were contaminated with C. jejuni. Differences were detected in the probability and level of contamination and the relative frequency of genotypes for individual poultry suppliers and humans. Some carcasses were contaminated with isolates belonging to more than one sequence type (ST), and there was evidence of both ubiquitous and supplier-associated strains, an epidemiological pattern not recognized yet in other countries. The common poultry STs were also common in human clinical cases, providing evidence that poultry is a major contributor to human infection. Both internationally rare genotypes, such as ST-3069 and ST-474, and common genotypes, such as ST-45 and ST-48, were identified in this study. The dominant human sequence type in New Zealand, ST-474, was found almost exclusively in isolates from one poultry supplier, which provided evidence that C. jejuni has a distinctive molecular epidemiology in this country. These results may be due in part to New Zealand's geographical isolation and its uniquely structured poultry industry.
Carvalheira, Ana; Casquete, Rocio; Silva, Joana; Teixeira, Paula
The prevalence and antibiotic resistance of Acinetobacter spp. from fifty samples of meat (chicken, turkey, beef and pork) were evaluated. Acinetobacter spp. was recovered from all samples and the clonal relatedness of 223 isolates identified to belong to the genus Acinetobacter was established by PFGE. A high genetic diversity was observed and 166 isolates from different samples, 141 representing different PFGE profiles, were further identified to the species level by rpoB gene sequencing. Thirteen distinct Acinetobacter species were identified among 156 isolates. The remaining ten isolates may represent three putatively novel species since rpoB sequence homologies with type strains of all available described Acinetobacter species, were <95%. The most common species was Acinetobacter guillouiae with a prevalence of 34.9%. However 18.7% of the strains belong to the Acinetobacter baumannii group (n=31) which include the species Acinetobacter baumannii (n=7), Acinetobacter pittii (n=12), Acinetobacter seifertii (n=8) and Acinetobacter nosocomialis (n=4) that are the species most frequently associated with nosocomial infections worldwide. In general, strains were resistant to some of the antimicrobials most frequently used to treat Acinetobacter infections such as piperacillin-tazobactam (64.9% of strains resistant), ceftazidime (43.5%), ciprofloxacin (42.9%), as well as to colistin (41.7%) and polymyxin B (35.1%), the last-resort drugs to treat infections caused by multidrug-resistant Acinetobacter. The percentage of resistant strains to trimethoprim-sulfamethoxazole, tetracycline, aminoglycosides (amikacin and tobramycin) and ampicillin-sulbactam was >10% (23.2%, 23.2%, 14.3%, 12.5%, 12.5%, respectively). However, resistances to meropenem, imipenem and minocycline were only sporadically observed (8.3%, 1.2% and 1.2%, respectively). Overall, 51.2% of the strains were considered as multidrug-resistant (MDR) and 9.6% as extensively drug-resistant (XDR). The prevalence
Hill, Stephen; Nowak, Eva; Edge, Thomas A.
This large-scale study compared incubation temperatures (37°C versus 42°C) to study the detection of thermophilic Campylobacter species, including Campylobacter jejuni, C. coli, and C. lari, in various surface water samples and bird fecal droppings around Hamilton Harbor, Lake Ontario. The putative culture isolates obtained from incubation temperatures of 37 and 42°C were confirmed by Campylobacter genus- and species-specific triplex PCR assays targeting the 16S rRNA gene and the 16S-23S rRNA gene internal transcribed spacer (ITS) region. A total of 759 water, wastewater, and bird fecal dropping samples were tested. Positive amplification reactions for the genus Campylobacter were found for 454 (60%) samples incubated at 37°C, compared to 258 (34%) samples incubated at 42°C. C. jejuni (16%) and C. lari (12%) were detected significantly more frequently at the 42°C incubation temperature than at 37°C (8% and 5%, respectively). In contrast, significantly higher rates of C. coli (14%) and other Campylobacter spp. (36%) were detected at the 37°C incubation temperature than at 42°C (8% and 7%, respectively). These results were consistent across surface water, wastewater, and bird fecal dropping samples. At times, Campylobacter spp. were recovered and detected at 37°C (3% for C. jejuni, 10% for C. coli, and 3% for C. lari) when the same samples incubated at 42°C were negative. A significantly higher rate of other Campylobacter spp. was detected only at 37°C (32%) than only at 42°C (3%). These results indicate that incubation temperature can significantly influence the culturability and detection of thermophilic and other fastidious Campylobacter spp. and that a comprehensive characterization of the Campylobacter spp. in surface water, wastewaters, or bird fecal droppings will require incubation at both 37 and 42°C. PMID:24077717
Three pathogens, Campylobacter, Salmonella, and Shiga toxin producing Escherichia coli (STEC), are leading causes of bacterial gastroenteritis in the United States and worldwide. For example, Campylobacter species are responsible for 17% of all hospitalizations related to illness, and although Campy...
Nagamine, Claude M; Shen, Zeli; Luong, Richard H; McKeon, Gabriel P; Ruby, Norman F; Fox, James G
We report the isolation of a novel helicobacter isolated from the caecum of the Siberian hamster (Phodopus sungorus). Sequence analysis showed 97% sequence similarity to Helicobacter ganmani. In addition, we report the co-infection of these Siberian hamsters with a Campylobacter sp. and a second Helicobacter sp. with 99% sequence similarity to Helicobacter sp. flexispira taxon 8 (Helicobacter bilis), a species isolated previously from patients with bacteraemia. Gross necropsy and histopathology did not reveal any overt pathological lesions of the liver and gastrointestinal tract that could be attributed to the Helicobacter or Campylobacter spp. infections. This is the first helicobacter to be identified in the Siberian hamster and the first report of co-infection of Helicobacter spp. and Campylobacter sp. in asymptomatic Siberian hamsters.
Shen, Zeli; Luong, Richard H.; McKeon, Gabriel P.; Ruby, Norman F.; Fox, James G.
We report the isolation of a novel helicobacter isolated from the caecum of the Siberian hamster (Phodopus sungorus). Sequence analysis showed 97 % sequence similarity to Helicobacter ganmani. In addition, we report the co-infection of these Siberian hamsters with a Campylobacter sp. and a second Helicobacter sp. with 99 % sequence similarity to Helicobacter sp. flexispira taxon 8 (Helicobacter bilis), a species isolated previously from patients with bacteraemia. Gross necropsy and histopathology did not reveal any overt pathological lesions of the liver and gastrointestinal tract that could be attributed to the Helicobacter or Campylobacter spp. infections. This is the first helicobacter to be identified in the Siberian hamster and the first report of co-infection of Helicobacter spp. and Campylobacter sp. in asymptomatic Siberian hamsters. PMID:25752854
Lin, Jun; Yan, Meiguan; Sahin, Orhan; Pereira, Sonia; Chang, Yun-Juan; Zhang, Qijing
In this work we conducted both in vitro and in vivo experiments to examine the development and mechanisms of erythromycin (Ery) resistance in Campylobacter jejuni and Campylobacter coli. In vitro plating revealed that both Campylobacter species had similar but low spontaneous mutation frequencies (3 x 10(-9) to <5.41 x 10(-10)) for Ery resistance. Chickens infected with C. jejuni or C. coli were subjected to single or multiple treatments with medicated water containing tylosin (0.53 g/liter), which transiently reduced the level of Campylobacter colonization but did not select for Ery-resistant (Ery(r)) mutants in the treated birds. However, when tylosin was given to the chickens in feed at a growth-promoting dose (0.05 g/kg feed), Ery(r) mutants emerged in the birds after prolonged exposure to the antibiotic. The vast majority of the in vitro- and in vivo-selected Campylobacter mutants with Ery MICs of 8 to 256 microg/ml lacked the known resistance-associated mutations in the 23S rRNA gene, while the highly resistant mutants (Ery MIC > 512 microg/ml) had the A2074G mutation in the 23S rRNA gene. Inactivation of CmeABC, a multidrug efflux pump, dramatically reduced the Ery MIC in all of the examined mutants regardless of the presence of the A2074G mutation. Together, these results reveal distinct features associated with Ery resistance development in Campylobacter, demonstrate the significant role of CmeABC in Ery resistance, and suggest that long-term use of a macrolide as a growth promoter selects for the emergence of Ery(r) Campylobacter in animal reservoirs.
Oishi, Akira; Murakami, Koichi; Etoh, Yoshiki; Sera, Nobuyuki; Horikawa, Kazumi
Recently, there has been a marked increase in the number of reports of fluoroquinolone-resistant Campylobacter jejuni and Campylobacter coli. The aim of this study was to evaluate the prevalence of antimicrobial resistance and its genetic determinants in Campylobacter species isolated from meat and human subjects in Fukuoka Prefecture, Japan. Between 2011 and 2013, 55 and 64 isolates were collected from meat (chicken meat and beef liver) and humans, respectively, in this prefecture. Antimicrobial susceptibility tests were conducted using the agar dilution method in accordance with the Clinical and Laboratory Standards Institute guidelines, using the following 11 antimicrobial agents : cephalexin, cefoxitin, nalidixic acid, ciprofloxacin, levofloxacin, tetracycline, minocycline, ampicillin, streptomycin, kanamycin and erythromycin. The susceptibility rates of the isolates to three quinolones (nalidixic acid, ciprofloxacin, levofloxacin) were 43.7%, 41.2%, 40.3%, respectively. All the isolates were multidrug resistant. Whereas 46.9%-51.6% of the human isolates were resistant to one or more of the quinolones, only 32.7%-34.5% of the meat isolates were resistant to one or more of the drugs. DNA sequencing showed that of the 50 quinolone resistant isolates 44 had position 86 isoleucine (Ile) substituted for threonine (Thr) in the GyrA protein (Thr86Ile). This amino acid substitution resulted from ACA to ATA and ACT to ATT mutations of codon 86 in C. jejuni and C. coli, respectively. Furthermore, two of the four C. jejuni isolates lacking the Thr86Ile mutation had combined Ser22Gly-Asn203Ser substitutions, while the remaining two isolates had combined Ser22Gly-Asn203Ser-Ala 206Val substitutions. These four isolates also had cmeABC sequences that differed from the quinolone sensitive C. jejuni ATCC33560(T) strain. In conclusion, C. jejuni and C. coli have relatively high quinolone resistance, and are resistant to other antibiotics. The new combination of amino acid
Arslan, S; Eyi, A; Özdemir, F
Pseudomonas spp. are aerobic, gram-negative bacteria that are recognized as major food spoilage microorganisms. A total of 32 (22.9%) Pseudomonas spp. from 140 homemade white cheese samples collected from the open-air public bazaar were isolated and characterized. The aim of the present study was to investigate the biochemical characteristics, the production of extracellular enzymes, slime and β-lactamase, and antimicrobial susceptibility of Pseudomonas spp. isolated from cheeses. The identified isolates including Pseudomonas pseudoalcaligenes, Pseudomonas alcaligenes, Pseudomonas aeruginosa, Pseudomonas fluorescens biovar V, and P. pseudoalcaligenes ssp. citrulli were found to produce extracellular enzymes, respectively: protease and lecithinase production (100%), and lipase activity (85.7, 42.9, 100, and 100%, and nonlipolytic, respectively). The isolates did not produce slime and had no detectable β-lactamase activity. The antimicrobial susceptibility of the isolates was tested using the disk diffusion method. Pseudomonas spp. had the highest resistance to penicillin G (100%), then sulphamethoxazole/trimethoprim (28.1%). However, all Pseudomonas spp. isolates were 100% susceptible to ceftazidime, ciprofloxacin, amikacin, gentamicin, and imipenem. Multidrug-resistance patterns were not observed among these isolates. In this study, Pseudomonas spp., exhibiting spoilage features, were isolated mainly from cheeses. Isolation of this organism from processed milk highlights the need to improve the hygienic practices. All of the stages in the milk processing chain during manufacturing have to be under control to achieve the quality and safety of dairy products.
Vegge, Christina S.; Jansen van Rensburg, Melissa J.; Rasmussen, Janus J.; Maiden, Martin C. J.; Johnsen, Lea G.; Danielsen, Morten; MacIntyre, Sheila; Ingmer, Hanne; Kelly, David J.
Isolates of the zoonotic pathogen Campylobacter are generally considered to be unable to metabolize glucose due to lack of key glycolytic enzymes. However, the Entner-Doudoroff (ED) pathway has been identified in Campylobacter jejuni subsp. doylei and a few C. coli isolates. A systematic search for ED pathway genes in a wide range of Campylobacter isolates and in the C. jejuni/coli PubMLST database revealed that 1.7% of >6,000 genomes encoded a complete ED pathway, including both C. jejuni and C. coli from diverse clinical, environmental and animal sources. In rich media, glucose significantly enhanced stationary phase survival of a set of ED-positive C. coli isolates. Unexpectedly, glucose massively promoted floating biofilm formation in some of these ED-positive isolates. Metabolic profiling by gas chromatography–mass spectrometry revealed distinct responses to glucose in a low biofilm strain (CV1257) compared to a high biofilm strain (B13117), consistent with preferential diversion of hexose-6-phosphate to polysaccharide in B13117. We conclude that while the ED pathway is rare amongst Campylobacter isolates causing human disease (the majority of which would be of agricultural origin), some glucose-utilizing isolates exhibit specific fitness advantages, including stationary-phase survival and biofilm production, highlighting key physiological benefits of this pathway in addition to energy conservation. PMID:27920773
Cabezas, Luisa; Calderon, Carolina; Medina, Luis Miguel; Bahamon, Isabela; Cardenas, Martha; Bernal, Adriana Jimena; Gonzalez, Andrés; Restrepo, Silvia
Endophytes are microorganisms that asymptomatically invade plant tissues. They can stimulate plant growth and/or provide defense against pathogen attacks through the production of secondary metabolites. Most endophyte species are still unknown, and because they may have several applications, the study of their metabolic capabilities is essential. We characterized 100 endophytes isolated from Espeletia spp., a genus unique to the paramo ecosystem, an extreme environment in the Andean mountain range. We evaluated the cellulolytic potential of these endophytes on the saccharification of the oil palm empty fruit bunch (OPEFB). The total cellulolytic activity was measured for each endophyte on filter paper (FPA). In addition, the specific carboxymethyl cellulase (CMCase), exoglucanase, and β-glucosidase activities were determined. We found four fungi positive for cellulases. Of these fungi, Penicillium glabrum had the highest cellulolytic activity after partial purification, with maximal CMCase, exoglucanase and β-glucosidase enzyme activities of 44.5, 48.3, and 0.45 U/ml, respectively. Our data showed that the bioprospection of fungi and the characterization of their enzymes may facilitate the process of biofuel production.
Macé, Sabrina; Haddad, Nabila; Zagorec, Monique; Tresse, Odile
Campylobacter is the leading cause of bacterial enteritis in the world. For this reason, this pathogen is widely studied. As a microaerophilic and capnophilic microorganism, this foodborne pathogen requires an atmosphere with reduced oxygen (O2) and elevated carbon dioxide (CO2) concentrations for its optimal growth in vitro. According to the procedure for Campylobacter spp. isolation and cultivation from food products and environmental samples, European and American standards recommend gas proportions of 5% O2 and 10% CO2, complemented with nitrogen (N2). However, in the literature, the reported proportion of O2 for microaerobic growth conditions of Campylobacter spp. can range from 2.5% to 15% and the reason for this variation is usually not explained. The use of different gas generating systems and media to detect and to grow Campylobacter from foodstuff and the lack of information about gas producing systems are the main sources of the loss of consistancy between data. In this review, the relevance, strengths and weaknesses of these methods and their impact on Campylobacter biology are discussed. In conclusion the minimum information concerning microaerobic gaseous atmospheres are suggested in order to better harmonize data obtained from research studies for a better understanding of Campylobacter features.
Whatmore, Adrian M; Davison, Nicholas; Cloeckaert, Axel; Al Dahouk, Sascha; Zygmunt, Michel S; Brew, Simon D; Perrett, Lorraine L; Koylass, Mark S; Vergnaud, Gilles; Quance, Christine; Scholz, Holger C; Dick, Edward J; Hubbard, Gene; Schlabritz-Loutsevitch, Natalia E
Two Gram-negative, non-motile, non-spore-forming coccoid bacteria (strains F8/08-60(T) and F8/08-61) isolated from clinical specimens obtained from baboons (Papio spp.) that had delivered stillborn offspring were subjected to a polyphasic taxonomic study. On the basis of 16S rRNA gene sequence similarities, both strains, which possessed identical sequences, were assigned to the genus Brucella. This placement was confirmed by extended multilocus sequence analysis (MLSA), where both strains possessed identical sequences, and whole-genome sequencing of a representative isolate. All of the above analyses suggested that the two strains represent a novel lineage within the genus Brucella. The strains also possessed a unique profile when subjected to the phenotyping approach classically used to separate species of the genus Brucella, reacting only with Brucella A monospecific antiserum, being sensitive to the dyes thionin and fuchsin, being lysed by bacteriophage Wb, Bk2 and Fi phage at routine test dilution (RTD) but only partially sensitive to bacteriophage Tb, and with no requirement for CO2 and no production of H2S but strong urease activity. Biochemical profiling revealed a pattern of enzyme activity and metabolic capabilities distinct from existing species of the genus Brucella. Molecular analysis of the omp2 locus genes showed that both strains had a novel combination of two highly similar omp2b gene copies. The two strains shared a unique fingerprint profile of the multiple-copy Brucella-specific element IS711. Like MLSA, a multilocus variable number of tandem repeat analysis (MLVA) showed that the isolates clustered together very closely, but represent a distinct group within the genus Brucella. Isolates F8/08-60(T) and F8/08-61 could be distinguished clearly from all known species of the genus Brucella and their biovars by both phenotypic and molecular properties. Therefore, by applying the species concept for the genus Brucella suggested by the ICSP
Nadeau, Eric; Messier, Serge; Quessy, Sylvain
The in vitro virulence properties of 197 temporally and geographically related Campylobacter isolates from chicken broilers and humans were compared. Comparisons of the virulence properties associated with genotypes and biotypes were made. All isolates adhered to, and 63% invaded, INT-407 cells, whereas 13% were cytotoxic for CHO cells. CHO cell-cytotoxic extracts were also cytotoxic for INT-407 cells, but the sensitivity for Vero cells was variable. The proportion of isolates demonstrating a high invasiveness potential (>1,000 CFU ml(-1)) or Vero cell cytotoxicity was significantly higher for human than for poultry isolates. Invasiveness was associated with Campylobacter jejuni isolates of biotypes 1 and 2, whereas CHO and INT-407 cell cytotoxicity was associated with C. jejuni isolates of biotypes 3 and 4. Cytotoxic isolates were also clustered according to pulsed-field gel electrophoresis profiles.
Halbert, Lisa W; Kaneene, John B; Linz, John; Mansfield, Linda S; Wilson, Dave; Ruegg, Pamela L; Warnick, Lorin D; Wells, Scott J; Fossler, Charles P; Campbell, Amy M; Geiger-Zwald, Angela M
Campylobacter is one of the most common causes of gastroenteritis and can be acquired through contact with farm animals or the consumption of raw milk. Because of concerns over the role of food-producing animals in the dissemination of antimicrobial resistance to humans, we evaluated the prevalence of antimicrobial resistance in Campylobacter isolates from dairy farms and the genetic mechanism conferring the observed resistance. Evaluation of antimicrobial resistance was completed on 912 isolates from conventional and 304 isolates from organic dairy farms to eight drugs (azithromycin, chloramphenicol, ciprofloxacin, clindamycin, erythromycin, gentamicin, nalidixic acid, and tetracycline) with microbroth dilution. Resistance to seven of eight drugs was very low and did not differ by farm type. However, tetracycline resistance was common in Campylobacter isolated from both organic and conventional dairy farms, with 48 and 58% of isolates affected, respectively. By multiplex PCR, we determined that tetracycline resistance was highly associated with the carriage of tetO in Campylobacter isolates (X2 = 124, P < 0.01, kappa = 0.86).
Riboldi, Gustavo Pelicioli; Frazzon, Jeverson; d’Azevedo, Pedro Alves; Frazzon, Ana Paula Guedes
Fifty-six Enterococcus spp. strains were isolated from foods in Southern Brazil, confirmed by PCR and classified as Enterococcus faecalis (27), Enterococcus faecium (23) and Enterococcus spp (6). Antimicrobial susceptibility tests showed resistance phenotypes to a range of antibiotics widely administrated in humans such as gentamycin, streptomycin, ampicillin and vancomycin. PMID:24031330
Ohishi, Takayuki; Aoki, Kotaro; Ishii, Yoshikazu; Usui, Masaru; Tamura, Yutaka; Kawanishi, Michiko; Ohnishi, Kenji; Tateda, Kazuhiro
In this research, we analyzed the main sequence types (ST) and ST complexes of human- and chicken-derived isolates of Campylobacter jejuni in Japan by using multilocus sequence typing (MLST). We also analyzed lipooligosaccharide biosynthesis locus classes (LOS locus classes) and the numbers of isolates carrying genes coding resistance factors against various antibiotics, and observed their relationships. ST-21 complex was the main ST complex in isolates from humans (n = 38) and chickens (n = 25). None of the isolates showed resistance to imipenem, chloramphenicol, or erythromycin. Few isolates were resistant to ampicillin and streptomycin (1.3%-15%), whereas many showed resistance to tetracycline, ciprofloxacin, and nalidixic acid (38%-48%). Among the ST-21 complex isolates, ST4526 was detected at a very high rate. Those isolates showed resistance to tetracycline and ciprofloxacin, and were susceptible to ampicillin. Among the chicken-derived isolates, 37 of the 38 isolates that showed resistance to ciprofloxacin and nalidixic acid had threonine to isoleucine amino acid substitution in GyrA at codon 86 (T86I). Among the human-derived isolates, 17 of the 47 isolates that showed resistance to ciprofloxacin and 16 of the 48 isolates that showed resistance to nalidixic acid did not have T86I amino acid mutations in GyrA. The human-derived ST-21 complex isolates were classified into LOS locus classes A, B, C, D, and E. The chicken-derived ST-21 complex isolates, with the exception of one isolate, were all classified into LOS locus classes C and D. Among chicken-derived isolates, the most prevalent was ST51 (ST-443 complex) (10 isolates) and all of those were LOS locus class E.
Wells, J G; Morris, G K
Recovery of Shigella spp. from fecal specimens transported in buffered glycerol saline and Cary-Blair media held at frozen, refrigerated, or room temperature was compared with recovery by direct plating of fecal specimens. Buffered glycerol saline was the better transport medium for the recovery of Shigella spp. Refrigerated or frozen transport temperatures were superior to room temperature for recovery from either medium.
Shivaji, S; Rao, N S; Saisree, L; Sheth, V; Reddy, G S; Bhargava, P M
Ten cultures of Pseudomonas spp. were established from soil samples collected in and around a lake in Antarctica. Based on their morphology, biochemical and physiological characteristics, and moles percent G + C of their DNA, they were identified as P. fluorescens, P. putida, and P. syringae. This is the first report on the identification of Pseudomonas spp. from continental Antarctica. PMID:2930174
Royden, A; Wedley, A; Merga, J Y; Rushton, S; Hald, B; Humphrey, T; Williams, N J
Campylobacter is the leading cause of bacterial diarrhoeal disease worldwide, with raw and undercooked poultry meat and products the primary source of infection. Colonization of broiler chicken flocks with Campylobacter has proved difficult to prevent, even with high levels of biosecurity. Dipteran flies are proven carriers of Campylobacter and their ingress into broiler houses may contribute to its transmission to broiler chickens. However, this has not been investigated in the UK. Campylobacter was cultured from 2195 flies collected from four UK broiler farms. Of flies cultured individually, 0·22% [2/902, 95% confidence interval (CI) 0-0·53] were positive by culture for Campylobacter spp. Additionally, 1293 flies were grouped by family and cultured in 127 batches: 4/127 (3·15%, 95% CI 0·11-6·19) from three broiler farms were positive for Campylobacter. Multilocus sequence typing of isolates demonstrated that the flies were carrying broiler-associated sequence types, responsible for human enteric illness. Malaise traps were used to survey the dipteran species diversity on study farms and also revealed up to 612 flies present around broiler-house ventilation inlets over a 2-h period. Therefore, despite the low prevalence of Campylobacter cultured from flies, the risk of transmission by this route may be high, particularly during summer when fly populations are greatest.
Jeffrey, J. S.; Hunter, A.; Atwill, E. R.
The objective of this study was to produce an economical, easy to prepare, field-suitable enrichment medium for detection of Campylobacter jejuni in small numbers. A semisolid aerobic enrichment medium was developed. Rates of recovery from inoculated medium, sterile swabs, and mixed cultures of C. jejuni and coliform bacteria were tested. PMID:10747165
INTRODUCTION: Significant interest in studying the lipooligosaccharide (LOS) of Campylobacter jejuni stemmed from its potential role in post-infection paralytic disorders. METHODS: In this study we present PCR screening of five LOS locus classes (A, B, C, D, and E), for a collection of 116 C. jeju...
Lévesque, Simon; Frost, Eric; Arbeit, Robert D; Michaud, Sophie
Molecular strain typing is essential for deciphering the epidemiology of Campylobacter jejuni infections. We applied two different methods, multilocus sequence typing (MLST) and analysis of the flaA short variable repeat (SVR), to 289 isolates (163 human, 56 chicken, 34 raw milk, and 36 environmental water isolates) collected in the province of Québec, Canada, over 3 years; in addition, the analysis included the pulsed-field gel electrophoresis (PFGE) typing results for a subset of 131 isolates studied previously. MLST defined 96 sequence types (STs) and 20 clonal complexes (CCs), including 49 STs (73 isolates, 25%) and 39 alleles not previously documented in an international database. The frequency of new STs was significantly higher among water isolates than among isolates from other sources (18/36 [50%] and 55/253 [22%], respectively; P < 0.001). Nine of the 10 most prevalent CCs included isolates from humans and at least one other source; five CCs comprised exclusively or mostly human and chicken isolates. However, water and milk were the predominant nonhuman sources among the remaining CCs, suggesting that sporadic C. jejuni infections in humans may frequently arise from sources other than chickens. All three typing systems were discriminatory (discriminatory index > 0.9). Among 131 isolates analyzed by PFGE, each of the 20 types represented by two or more isolates corresponded to a single CC. In contrast, among the 14 most prevalent types detected by analysis of the flaA SVR (5 to 27 isolates each), 8 (57%) included isolates that represented multiple different CCs. The basis for these discordant results was uncertain. Antimicrobial resistance was randomly distributed among the CCs and appeared to be more closely related to the source of an isolate than its genotype. Although MLST is labor-intensive and expensive, it remains the single best method for the genotyping of C. jejuni isolates and deciphering the epidemiologic relationships among isolates.
Maschio, Vinicius José; Corção, Gertrudes; Rott, Marilise Brittes
Acanthamoeba is a “Trojan horse” of the microbial world. The aim of this study was to identify the presence of Pseudomonas as an amoeba-resistant microorganism in 12 isolates of Acanthamoeba. All isolates showed the genus Pseudomonas spp. as amoeba-resistant microorganisms. Thus, one can see that the Acanthamoeba isolates studied are hosts of Pseudomonas. PMID:25651331
José Maschio, Vinicius; Corção, Gertrudes; Rott, Marilise Brittes
Acanthamoeba is a "Trojan horse" of the microbial world. The aim of this study was to identify the presence of Pseudomonas as an amoeba-resistant microorganism in 12 isolates of Acanthamoeba. All isolates showed the genus Pseudomonas spp. as amoeba-resistant microorganisms. Thus, one can see that the Acanthamoeba isolates studied are hosts of Pseudomonas.
SOMROOP, Srinuan; HATANAKA, Noritoshi; AWASTHI, Sharda Prasad; OKUNO, Kentaro; ASAKURA, Masahiro; HINENOYA, Atsushi; YAMASAKI, Shinji
Cytolethal distending toxin (CDT) consisting of CdtA, CdtB and CdtC has been reported to be a possible virulence factor of campylobacters including Campylobacter upsaliensis. In our previous study, the cdtB gene-based PCR-restriction fragment length polymorphism (RFLP) assay for detection and differentiation of 7 Campylobacter species yielded 3 different RFLP patterns (Cu-I to Cu-III). In this study, entire cdt (Cucdt) genes of each pattern were sequenced to see whether there are any differences in cdt genes, its amino acid sequences and biological activity of CuCDT. We found that all 3 representative strains harbor the entire Cucdt genes and homology between prototype and newly determined Cucdt genes was 94 to 98% with cdtA, 93 to 94% with cdtB and 92 to 93% with cdtC, while that between amino acids of CuCDT was 95 to 99% with CdtA, 97 to 98% with CdtB and 92 to 93% with CdtC. Furthermore, CDT activity produced by C. upsaliensis strains was examined by cytotoxicity assay with HeLa cells. Interestingly, C. upsaliensis produced 64 to 2,340 times higher CDT titer in comparison to other campylobacters did. In addition, Cu-III showed 64 times higher CDT titer than Cu-II, although CDT production level was almost the same by western blotting. These data suggest that CDT produced by C. upsaliensis might contribute more to human diseases in comparison to that produced by other campylobacters and Cu-III CDT seems to be more toxic to HeLa cells in comparison to Cu-I and Cu-II CDTs. PMID:28202878
Quantitative risk assessment of thermophilic Campylobacter spp. and cross-contamination during handling of raw broiler chickens evaluating strategies at the producer level to reduce human campylobacteriosis in Sweden.
Lindqvist, Roland; Lindblad, Mats
Campylobacter is a major bacterial cause of infectious diarrheal illness in Sweden and in many other countries. Handling and consumption of chicken has been identified as important risk factors. The purpose of the present study was to use data from a national baseline study of thermophilic Campylobacter spp. in raw Swedish broiler chickens in order to evaluate some risk management strategies and the frequency of consumer mishandling, i.e., handling leading to possible cross-contamination. A probabilistic model describing variability but not uncertainty was developed in Excel and @Risk. The output of the model was the probability of illness per handling if the chicken was mishandled. Uncertainty was evaluated by performing repeated simulations and substituting model parameters, distributions and software (Analytica). The effect of uncertainty was within a factor of 3.2 compared to the baseline scenario. For Campylobacter spp. prevalence but not concentration, there was a one-to-one relation with risk. The effect of a 100-fold reduction in the levels of Campylobacter spp. on raw chicken reduced the risk by a factor of 12 (fresh chicken) to 30 (frozen chicken). Highly-contaminated carcasses contributed most to risk and it was estimated that by limiting the contamination to less than 4 log CFU per carcass, the risk would be reduced to less than 17% of the baseline scenario. Diverting all positive flocks to freezing was estimated to result in 43% as many cases as the baseline. The second best diversion option (54% of baseline cases) was to direct all chickens from the two worst groups of producers, in terms of percentages of positive flocks delivered, to freezing. The improvement of using diverting was estimated to correspond to between 5 to 767 fewer reported cases for the different strategies depending on the assumptions of the proportion of reported cases (1 to 50%) caused by Campylobacter spp. from Swedish chicken. The estimated proportion of consumer mishandlings
Zaidi, Mussaret B; McDermott, Patrick F; Campos, Freddy D; Chim, Rodolfo; Leon, Magda; Vazquez, Gabriela; Figueroa, Gloria; Lopez, Estela; Contreras, Jesus; Estrada-Garcia, Teresa
We describe prevalence and antimicrobial susceptibility results for thermophilic Campylobacter isolates collected from humans, food, and food-animals in an integrated food chain surveillance network in Mexico. From 2003 to 2006, stool samples were collected from children with diarrhea at state sentinel hospitals. Concurrently, fecal samples from asymptomatic children in kindergartens, as well as raw chicken, pork and beef from retail outlets, and food-animal intestines from slaughterhouses were all collected in 65 cities from four different states. C. jejuni was identified with a standardized hippurate test. Hippurate negative, indoxyl acetate positive isolates were classified as Campylobacter spp. Susceptibility testing was performed by agar dilution according to Clinical and Laboratory Standards Institute guidelines. A total of 1,259 C. jejuni and 1,797 Campylobacter spp. isolates were recovered from 11,811 samples. Chicken was significantly more contaminated for both intestinal samples (93.6%) and meat products (58.3%), compared with swine (71.4%)/pork (14.6%) samples, and cattle (25.1%)/beef (5.3%) samples (p<0.001). Campylobacter was recovered from 5.1% of children with diarrhea and from 3.2% of asymptomatic children. Chicken was significantly more likely to harbor ciprofloxacin-resistant C. jejuni (85.8%) than swine (62.5%, OR=3.6), cattle (39.8%, OR=9.3), or humans (58.2%, OR=4.4). No significant differences were found for ciprofloxacin-resistant Campylobacter spp. among food-animals, but the rate in food-animals was significantly higher than in humans (84% vs. 56.7%, OR=4.0). Swine was significantly more likely to harbor erythromycin-resistant C. jejuni (14.8%) than chicken (3.5%, OR=4.9), cattle (1.8%, OR=9.3), or humans (3.0%, OR=5.7), and was associated with higher rates of erythromycin-resistant Campylobacter spp. (41.9%) than chicken (10.5%, OR=6.1) and humans (11.9%, OR=5.3). The high resistance rates to ciprofloxacin preclude the use of
Fitzgerald, Collette; Tu, Zheng Chao; Patrick, Mary; Stiles, Tracy; Lawson, Andy J; Santovenia, Monica; Gilbert, Maarten J; van Bergen, Marcel; Joyce, Kevin; Pruckler, Janet; Stroika, Steven; Duim, Birgitta; Miller, William G; Loparev, Vladimir; Sinnige, Jan C; Fields, Patricia I; Tauxe, Robert V; Blaser, Martin J; Wagenaar, Jaap A
A polyphasic study was undertaken to determine the taxonomic position of 13 Campylobacter fetus-like strains from humans (n = 8) and reptiles (n = 5). The results of matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) MS and genomic data from sap analysis, 16S rRNA gene and hsp60 sequence comparison, pulsed-field gel electrophoresis, amplified fragment length polymorphism analysis, DNA-DNA hybridization and whole genome sequencing demonstrated that these strains are closely related to C. fetus but clearly differentiated from recognized subspecies of C. fetus. Therefore, this unique cluster of 13 strains represents a novel subspecies within the species C. fetus, for which the name Campylobacter fetus subsp. testudinum subsp. nov. is proposed, with strain 03-427(T) ( = ATCC BAA-2539(T) = LMG 27499(T)) as the type strain. Although this novel taxon could not be differentiated from C. fetus subsp. fetus and C. fetus subsp. venerealis using conventional phenotypic tests, MALDI-TOF MS revealed the presence of multiple phenotypic biomarkers which distinguish Campylobacter fetus subsp. testudinum subsp. nov. from recognized subspecies of C. fetus.
Mund, Norah Lynn-Anne; Masanta, Wycliffe Omurwa; Goldschmidt, Anne-Marie; Lugert, Raimond; Groß, Uwe; Zautner, Andreas E.
Campylobacter jejuni’s flagellar locomotion is controlled by eleven chemoreceptors. Assessment of the distribution of the relevant chemoreceptor genes in the C. jejuni genomes deposited in the National Center for Biotechnology Information (NCBI) database led to the identification of two previously unknown tlp genes and a tlp5 pseudogene. These two chemoreceptor genes share the same locus in the C. jejuni genome with tlp4 and tlp11, but the gene region encoding the periplasmic ligand binding domain differs significantly from other chemoreceptor genes. Hence, they were named tlp12 and tlp13. Consequently, it was of interest to study their distribution in C. jejuni subpopulations of different clonality, and their cooccurrence with the eleven previously reported chemoreceptor genes. Therefore, the presence of all tlp genes was detected by polymerase chain reaction (PCR) in 292 multilocus sequence typing (MLST)-typed C. jejuni isolates from different hosts. The findings show interesting trends: Tlp4, tlp11, tlp12, and tlp13 appeared to be mutually exclusive and cooccur in a minor subset of isolates. Tlp4 was found to be present in only 33.56% of all tested isolates and was significantly less often detected in turkey isolates. Tlp11 was tested positive in only 17.8% of the isolates, while tlp12 was detected in 29.5% of all isolates, and tlp13 was found to be present in 38.7%. PMID:27766165
Jamali, Hossein; Radmehr, Behrad
The aims of this study were to determine the prevalence, characteristics and antimicrobial resistance of Listeria spp. isolated from bovine clinical mastitis in Iran. Listeria spp. were detected in 21/207 bovine mastitic milk samples from dairy farms in Iran, comprising L. monocytogenes (n=17), L. innocua (n=3) and L. ivanovii (n=1). L. monocytogenes isolates were grouped into serogroups '4b, 4d, 4e', '1/2a, 3a', '1/2b, 3b, 7' and '1/2c, 3c'; all harboured inlA, inlC and inlJ virulence genes. Listeria spp. were most frequently resistant to penicillin G (14/21 isolates, 66.7%) and tetracyclines (11/21 isolates, 52.4%).
For approximately three decades, the genus Campylobacter has had increased focus as a threat to food safety, due to the rise in enteritis in humans caused by consumption or handling of foods contaminated with the organism. For this reason, numerous research studies have been conducted and books wri...
During 2009 and 2010, 45 isolates of Rhizoctonia spp. were recovered from onion bulb crops in the semi-arid Columbia Basin of Oregon and Washington, in which patches of severely stunted onion plants developed following rotation with winter cereal cover crops. Characterization of isolates recovered f...
Aims: The aims of this study were firstly to compare five published methods for the isolation of Arcobacter spp. from animal faeces in order to determine the most sensitive and specific method. Secondly, we analyzed the resulting isolates by multi-locus sequence typing (MLST) in order to investiga...
Cody, Alison J; McCarthy, Noel D; Bray, James E; Wimalarathna, Helen M L; Colles, Frances M; Jansen van Rensburg, Melissa J; Dingle, Kate E; Waldenström, Jonas; Maiden, Martin C J
The contribution of wild birds as a source of human campylobacteriosis was investigated in Oxfordshire, United Kingdom (UK) over a 10 year period. The probable origin of human Campylobacter jejuni genotypes, as described by multilocus sequence typing, was estimated by comparison with reference populations of isolates from farm animals and five wild bird families, using the STRUCTURE algorithm. Wild bird-attributed isolates accounted for between 476 (2.1%) and 543 (3.5%) cases annually. This proportion did not vary significantly by study year (P = 0.934) but varied seasonally, with wild bird-attributed genotypes comprising a greater proportion of isolates during warmer compared with cooler months (P = 0.003). The highest proportion of wild bird-attributed illness occurred in August (P < 0.001), with a significantly lower proportion in November (P = 0.018). Among genotypes attributed to specific groups of wild birds, seasonality was most apparent for Turdidae-attributed isolates, which were absent during cooler, winter months. This study is consistent with some wild bird species representing a persistent source of campylobacteriosis, and contributing a distinctive seasonal pattern to disease burden. If Oxfordshire is representative of the UK as a whole in this respect, these data suggest that the national burden of wild bird-attributed isolates could be in the order of 10,000 annually.
Levrè, E; Valentini, P; Brunetti, M; Sacchelli, F
Domestic and wild animals have been always considered very important as reservoir of agents of human infections. Particularly birds, because of their great mobility from a continent to another or within the limits of the same ecosystem may transfer pathogenic micro-organisms. The present survey was undertaken in order to evaluate the presence of Campylobacter, Yersinia and Salmonella in migratory and permanent birds. During the period October 1986 to March 1988 intestinal loops were collected from a total of 217 birds representing 17 different species shot, during hunting seasons, in the inland of Versilia in the district of Lucca. Each sample was divided into three parts and examined for the presence of Campylobacter, Yersinia and Salmonella. Campylobacter was isolated from 74 of the 217 birds examined (34.10%). Yersinia was recovered from 26 birds (11.98%), while only 8 birds (3.68%) harboured Salmonella. Most of the samples carried only one of the three bacterial genera investigated while 9 harboured at the same time Yersinia and Campylobacter, 1 Salmonella and Campylobacter, and 1 Salmonella, Yersinia and Campylobacter. Campylobacter spp.: On the ground of Lior's biotyping scheme, the 74 strains, isolated from 16 of the 17 species of birds examined, have assigned to three biochemically different species. C. coli was the most commonly isolated followed by C. jejuni and C. laridis. Of the 54 isolates of C. coli 30 belonged to biotype I and 24 to biotype II. 19 C. jejuni organisms were differentiated into 5 belonging to biotype I and 14 to biotype II. The only C. laridis isolated belonged to biotype I. Yersinia spp.: 37 strains belonging to the genus Yersinia were isolated from 26 birds. In 10 samples different types of Yersinia were identified. Most of the strains could be ascribed to Yersinia enterocolitica (28 strains), 3 to Yersinia frederiksenii, 3 to Yersinia intermedia and 1 to Yersinia pseudotuberculosis. 2 strains were identified as atypical Yersinia
Poly, Frédéric; Serichantalergs, Oralak; Kuroiwa, Janelle; Pootong, Piyarat; Mason, Carl; Guerry, Patricia; Parker, Craig T.
Campylobacter jejuni produces a polysaccharide capsule that is the major determinant of the Penner serotyping scheme. This passive slide agglutination typing system was developed in the early 1980’s and was recognized for over two decades as the gold standard for C. jejuni typing. A preliminary multiplex PCR technique covering 17 serotypes was previously developed in order to replace this classic serotyping scheme. Here we report the completion of the multiplex PCR technology that is able to identify all the 47 Penner serotypes types known for C. jejuni. The number of capsule types represented within the 47 serotypes is 35. We have applied this method to a collection of 996 clinical isolates from Thailand, Cambodia and Nepal and were able to successfully determine capsule types of 98% of these. PMID:26630669
Crespo, M D; Altermann, E; Olson, J; Miller, W G; Chandrashekhar, K; Kathariou, S
In Campylobacter spp., resistance to the antimicrobials kanamycin and tetracycline is frequently associated with plasmid-borne genes. However, relatively few plasmids of Campylobacter jejuni have been fully characterized to date. A novel plasmid (p11601MD; 44,095nt) harboring tet(O) was identified in C. jejuni strain 11601MD, which was isolated from the jejunum of a turkey produced conventionally in North Carolina. Analysis of the p11601MD sequence revealed the presence of a high-GC content cassette with four genes that included tet(O) and a putative aminoglycoside transferase gene (aphA-3) highly similar to kanamycin resistance determinants. Several genes putatively involved in conjugative transfer were also identified on the plasmid. These findings will contribute to a better understanding of the distribution of potentially self-mobilizing plasmids harboring antibiotic resistance determinants in Campylobacter spp. from turkeys and other sources.
Campylobacter spp. are microaerophilic and capnophilic microorganisms. Methods to increase exposure of Campylobacter to appropriate microaerobic conditions could theoretically improve recovery of stressed cells. The porous nature of a sponge greatly increases the sample surface area exposed to mic...
Mąka, Łukasz; Maćkiw, Elżbieta; Ścieżyńska, Halina; Modzelewska, Magdalena; Popowska, Magdalena
Antimicrobial resistance of pathogenic bacteria, including Salmonella spp., is an emerging problem of food safety. Antimicrobial use can result in selection of resistant organisms. The food chain is considered a route of transmission of resistant pathogens to humans. In many European countries, sulfonamides are one of the most commonly used antimicrobials. The aim of our investigation was to assess the prevalence of sul genes and plasmid occurrence among sulfonamide-resistant Salmonella spp. Eighty-four sulfonamide-resistant isolates were collected in 2008 and 2013 from retail products in Poland. Minimal inhibitory concentration of all of these isolates was ≥1024 μg/mL. Resistant isolates were tested for the presence of sul1, sul2, sul3, and int1 genes by using multiplex polymerase chain reaction. In total, 44.0% (37/84) isolates carried the sul1 gene, 46.4% (39/84) were sul2 positive, while the sul3 gene was not detected in any of the sulfonamide-resistant isolates tested. It was found that 3.6% (3/84) of resistant Salmonella spp. contained sul1, sul2, and intI genes. All 33 intI-positive isolates carried the sul1 gene. Eleven of the sulfonamide-resistant isolates were negative for all the sul genes. Most of the sulfonamide-resistant Salmonella spp. harbored plasmids; only in eight isolates were no plasmids detected. Generally, the size of the plasmids ranged from approximately 2 kb to ≥90 kb. Our results revealed a relatively a high prevalence of sulfonamides-resistant Salmonella spp. isolated from retail food. Additionally, we have detected a high dissemination of plasmids and class 1 integrons that may enhance the spread of resistance genes in the food chain.
Introduction: Campylobacter spp. are considered to be a leading bacterial etiologic agent of acute food-borne gastroenteritis among human populations. Epithelial cell invasion is hypothesized to be necessary for human infection and cell invasion assays have been utilized to demonstrate that distinc...
Patchanee, Prapas; Chokboonmongkol, Chomporn; Zessin, Karl-Hans; Alter, Thomas; Pornaem, Sarinya; Chokesajjawatee, Nipa
We compared rapid fingerprinting using repetitive sequence-based PCR (rep-PCR) for subtyping Campylobacter jejuni isolates to the widely used multilocus sequence typing (MLST). Representative C. jejuni isolates (n = 16) from broilers were analyzed using MLST and rep-PCR. Both techniques demonstrated an equal discriminatory power of 0.8917, and 9 subgroups were identified. Clonal identification of all 16 isolates was identical for both techniques. The rep-PCR as described in this study may be used as a rapid and cost-effective alternative for subtyping of C. jejuni isolates, or as an effective screening tool in large epidemiological studies.
Xu, Jiancheng; Wang, Liqiang; Wang, Kai; Zhou, Qi
This study was to investigate the antimicrobial resistance of Enterococcus spp. isolated in 8 consecutive years in the First Bethune Hospital. Disk diffusion test was used to study the antimicrobial resistance. The data were analyzed by WHONET 5 software according to Clinical and Laboratory Standards Institute (CLSI). Most of 1446 strains of Enterococcus spp. were collected from urine 640 (44.3%), sputum 315 (21.8%), secretions and pus 265 (18.3%) during the past 8 years. The rates of high-level aminoglycoside resistance in Enterococcus faecalis and Enterococcus faecium were 57.4%∼75.9% and 69.0%∼93.8% during the past 8 years, respectively. No Enterococcus spp. was resistant to vancomycin. The antimicrobial resistance of Enterococcus spp. had increased in recent 8 years. The change of the antimicrobial resistance should be investigated in order to direct rational drug usage in the clinic and prevent bacterial strain of drug resistance from being transmitted.
Queiroz Júnior, Luiz de Pádua; de Camargo, Zoilo Pires; Tadano, Tomoko; Rodrigues, Anderson Messias; Takarara, Doracilde Terumi; Gegembauer, Gregory; Araujo, Leticia Mendes; Hahn, Rosane Christine
Clinical Paracoccidioides spp. isolates from patients with paracoccidioidomycosis (PCM) in Mato Grosso, Brazil exhibit different patterns of serologic reactivity. The results observed for reactions of radial immunodiffusion against the commonly used exoantigens containing a 43-kDa glycoprotein (gp43) suggest that this fungus exhibits major antigenic variability by geographic region. There is a phylogenetic gap between Paracoccidioides spp. isolates among different regions of Latin America. In particular, those from the central region of Brazil (i.e. Mato Grosso state) exhibit a lower rate of genetic similarity. We aimed at investigating the phylogenetic classification of clinical isolates of Paracoccidioides spp. in Central Brazil and the different antigenic profiles that produce. Exoantigens were obtained from five clinical isolates: two P. brasiliensis (Pb166 and Pb2880) and three P. lutzii (PL2875, PL9840, and PL2912). The protein/glycoprotein profiles of P. lutzii exoantigens were different from each other. Isolate PL9840 exhibited the most distinct bands, and isolates PL2875 and PL2912 exhibited more diffuse bands and a very intense band between 50 and 60 kDa. P. brasiliensis isolates had similar protein profiles, exhibiting a low-intensity band at 220 kDa and a diffuse band between 50 and 60 kDa. P. lutzii isolates exhibit high species-specific antigen variability, which we have already been assessed in proteomic studies.
Adzitey, Frederick; Huda, Nurul; Ali, Gulam Rusul Rahmat
Campylobacter, Salmonella, and Listeria monocytogenes are important bacterial pathogens associated with gastroenteritis. The consumption of poultry meat and their products is considered as a major and leading source of human infection. While surveys of chicken meat and products, and its association with foodborne pathogens are widely available, such information on ducks is scarce. This survey examines the prevalence and antibiotic resistance of Campylobacter, Salmonella and L. monocytogenes isolated from ducks. Data obtained from key surveys are summarized. The observed prevalence of these pathogens and their resistance to various antibiotics varies from one study to the other. The mean prevalence (and range means from individual surveys) are duck 53.0% (0.0-83.3%), duck meat and parts 31.6% (12.5-45.8%), and duck rearing and processing environment 94.4% (92.0-96.7%) for Campylobacter spp. For Salmonella spp., the mean prevalence data are duck 19.9% (3.3-56.9%), duck meat and parts 28.4% (4.4-75.6%), duck egg, shell, and content 17.5% (0-4.17%), and duck rearing and processing environment 32.5% (10.5-82.6%). Studies on the prevalence and antibiotic resistance of L. monocytogenes in ducks are by far very rare compared to Campylobacter and Salmonella, although ducks have been noted to be a potential source for these foodborne pathogens. From our survey, ducks were more frequently contaminated with Campylobacter than Salmonella. Campylobacter and Salmonella spp. also exhibited varying resistance to multiple antibiotics.
Furuhata, Katsunori; Ishizaki, Naoto; Sogawa, Kazuyuki; Kawakami, Yasushi; Lee, Shin-Ichi; Sato, Masahiro; Fukuyama, Masafumi
From May 2014 to February 2015, 319 university students (male, n=173; female n=146) of 18 to 24 years of age who carried mobile phones or computer tablets were selected as subjects. Staphylococcus spp. were detected in 101 of 319 samples (31.7%). In the present study, 11 strains of S. aureus were isolated and identified, not all of which were methicillin-resistant Staphylococcus aureus (MRSA). Overall, 14 species were identified, with 11 strains (10.9%) of S. xylosus being isolated at the highest frequency. Following this were eight strains (7.9%) of S. cohnii and seven strains (6.9%) each of S. capitis and S. haemolyticus. Staphylococcus spp. isolation was performed with bacterial samples obtained from the mobile phones of 22 specific subjects (males, n=12; females, n=10). Staphylococcus spp. isolation was performed on days -1, 7 and 30 of the experiment. Staphylococcus spp. were positively detected one or more times in 12 subjects (54.5%). In one subject (8.3%), all three tests were positive. Furthermore, two tests were positive in three (25.0%). In the eight remaining subjects (66.7%) Staphylococcus spp. were detected only once. For the three abovementioned tests, we investigated the pulsed-field gel electrophoresis (PFGE) patterns of the strains derived from the mobile phone and from the fingers of three subjects in whom the same bacterial species were isolated twice. From the cases with similarities between strains derived from the fingers and the mobile phones and cases, with consistency in the strains derived from the mobile phone at different times, commonality was observed in the strains derived from the fingers and mobile phones along with chronological uniformity in the strains derived from the mobile phones. A total of 101 Staphylococcus spp. strains were isolated from mobile phones. According to drug susceptibility tests, 99 strains (98.0%) were found to have some degree of resistance to drugs (excluding one strain each of S. aureus and S. haemolyticus
Brandão, Marcelo Luiz Lima; Umeda, Natália Scudeller; Jackson, Emily; Forsythe, Stephen James; de Filippis, Ivano
Several Cronobacter species are opportunistic pathogens that cause infections in humans. The aim of this study was to detect Cronobacter spp. from 90 samples of retail foods in Brazil, and characterize the strains by phenotypic tests, molecular assays and antibiotic susceptibility. Three isolation methodologies were evaluated using different selective enrichments and the isolates were identified using Vitek 2.0, PCRs protocols, fusA allele sequencing and multilocus sequence typing (MLST). Thirty-eight samples (42.2%) contained Cronobacter spp., and the highest percentage was found in flours (66.7%, 20/30), followed by spices and herbs (36.7%, 11/30), and cereal mixes for children (23.3%, 7/30). The 45 isolates included four species: C. sakazakii (n = 37), C. malonaticus (n = 3), C. dublinensis (n = 3), and C. muytjensii (n = 2); that presented 20 different fusA alleles. MLST analysis revealed 32 sequence types (STs), 13 of which were newly identified. All strains were sensitive to all antibiotics (n = 10) tested. The combination of CSB/v enrichment with DFI plating was considered the most efficient for Cronobacter spp. isolation. This study revealed the presence of Cronobacter spp. in foods commercialized in Brazil and the isolates showed a high diversity after MLST analysis and included two strains of the C. sakazakii ST4 neonatal meningitic pathovar.
Soler, Carla; Esteban, J Guillermo; Jiménez, Ricardo; Mañes, Jordi; Soriano, José Miguel
Introducción: la transmisión de patógenos por insectos es una creciente preocupación para la salud pública. Más concretamente, las moscas son conocidas por ser capaces de transmitir el agente infeccioso mecánicamente. Objetivo: el presente trabajo muestra un estudio en los servicios de restauración en los que se aisló por primera vez en la literatura Megaselia spp, detectándose la presencia de microorganismos en estas moscas. Método: se basa en análisis microbiológicos y entomológicos. Resultados: la presencia de aerobios mesófilos y Enterobacteriaceae se han encontrado en todas las muestras, superando los límites establecidos en el 41,7% (5/12) para las bacterias aerobias mesófilas y el 66,7% (8/12) para Enterobacteriaceae. Por otra parte, en el 25 y 66,7% de las moscas analizadas se detectó la presencia de Escherichia coli y Staphylococcus aureus, respectivamente. Conclusiones: hay un binomio entre la presencia de microorganismos y Megaselia spp., lo que demuestra la importancia de mantener una vigilancia más estricta en las medidas higiénico-sanitarias en los servicios de restauración.
Kasai, J; Ando, F; Kuwashima, M
Legionella spp., the causative organism of legionnaires' disease, were isolated from more than 80% of water samples in cooling towers before washing. Therefore, we evaluated the effect of microbicide treatment of cooling tower water on Legionella spp., other bacteria and protozoa. 2-Bromo-2-nitropane-1,3-dial, 2,4-dibromo-5,5-dimethylhydantoin or silver nitrate-treated silica gel was added to cooling tower water. The isolation rate of Legionella spp. in the cooling tower water was 50% after microbiocide treatment with 2-bromo-2-nitropane-1,3-dial being the most effective. The microbicide treatment had no effect on other bacteria or protozoa. These findings indicated the importance of regular washing and water exchange of cooling tower water with microbicide treatment.
Ziech, Rosangela Estel; Lampugnani, Camila; Perin, Ana Paula; Sereno, Mallu Jagnow; Sfaciotte, Ricardo Antônio Pilegi; Viana, Cibeli; Soares, Vanessa Mendonça; Pinto, José Paes de Almeida Nogueira; Bersot, Luciano dos Santos
The aim of this study was to investigate the occurrence of multidrug-resistant, extended spectrum beta-lactamase (ESBL) producing Salmonella spp. isolated from conveyor belts of broiler cutting rooms in Brazilian broiler processing plants. Ninety-eight strains of Salmonella spp. were analyzed. Multidrug resistance was determined by the disk diffusion test and the susceptibility of the isolated bacteria was evaluated against 18 antimicrobials from seven different classes. The double disk diffusion test was used to evaluate ESBL production. Of the 98 strains tested, 84 were multidrug resistant. The highest rates of resistance were against nalidixic acid (95%), tetracycline (91%), and the beta-lactams: ampicillin and cefachlor (45%), followed by streptomycin and gentamicin with 19% and 15% of strain resistance, respectively. By contrast, 97% of the strains were sensitive to chloramphenicol. 45% of the strains were positive for the presence of ESBL activity. In this study, high rates of multidrug resistance and ESBL production were observed in Salmonella spp.
Elvers, Karen T.; Helps, Christopher R.; Wassenaar, Trudy M.; Allen, Vivien M.; Newell, Diane G.
The identification of sites resulting in cross-contamination of poultry flocks in the abattoir and determination of the survival and persistence of campylobacters at these sites are essential for the development of intervention strategies aimed at reducing the microbial burden on poultry at retail. A novel molecule-based method, using strain- and genus-specific oligonucleotide probes, was developed to detect and enumerate specific campylobacter strains in mixed populations. Strain-specific oligonucleotide probes were designed for the short variable regions (SVR) of the flaA gene in individual Campylobacter jejuni strains. A 16S rRNA Campylobacter genus-specific probe was also used. Both types of probes were used to investigate populations of campylobacters by colony lift hybridization. The specificity and proof of principle of the method were tested using strains with closely related SVR sequences and mixtures of these strains. Colony lifts of campylobacters were hybridized sequentially with up to two labeled strain-specific probes, followed by the generic 16S rRNA probe. SVR probes were highly specific, differentiating down to 1 nucleotide in the target sequence, and were sufficiently sensitive to detect colonies of a single strain in a mixed population. The 16S rRNA probe detected all Campylobacter spp. tested but not closely related species, such as Arcobacter skirrowi and Helicobacter pullorum. Preliminary field studies demonstrated the application of this technique to target strains isolated from poultry transport crate wash tank water. This method is quantitative, sensitive, and highly specific and allows the identification and enumeration of selected strains among all of the campylobacters in environmental samples. PMID:18281428
Campylobacter is a leading cause of foodborne illness worldwide. It is common in poultry, and human infections are often associated with consumption of contaminated poultry products. One strategy to reduce Campylobacter colonization in poultry is by using oral probiotics. Unfortunately, oral probiot...
Hernández-Domínguez, C; Guzmán-Franco, A W; Carrillo-Benítez, M G; Alatorre-Rosas, R; Rodríguez-Leyva, E; Villanueva-Jiménez, J A
Spittlebugs from the genus Aeneolamia are important pests of sugarcane. Although the use of the entomopathogenic fungus Metarhizum anisopliae s.l. for control of this pest is becoming more common in Mexico, fundamental information regarding M. anisopliae in sugarcane plantations is practically non-existent. Using phylogenetic analysis, we determined the specific diversity of Metarhizium spp. infecting adult spittlebugs in sugarcane plantations from four Mexican states. We obtained 29 isolates of M. anisopliae s.str. Haplotype network analysis revealed the existence of eight haplotypes. Eight selected isolates, representing the four Mexican states, were grown at different temperatures in vitro; isolates from Oaxaca achieved the greatest growth followed by isolates from Veracruz, San Luis Potosi and Tabasco. No relationship was found between in vitro growth and haplotype diversity. Our results represent a significant contribution to the better understanding of the ecology of Metarhizum spp. in the sugarcane agroecosystem.
Kanan, T A; Abdulla, Z A
All 250 children presenting with diarrhoea at 2 teaching hospitals in Mosul, Iraq over a 9-month period were studied for the presence of Yersinia spp. in stools by cold-enrichment culture at 4 degrees C for 21 days. Pathogenicity of the isolated Yersinia was determined. Antibodies to Y. enterocolitica were raised for rapid Yersinia detection in the stool. Yersinia spp. were isolated from the stools of only 4 patients; 3 isolates were identified as Y. enterocolitica and 1 was Y. pseudotuberculosis. The blood culture was also positive for Y. enterocolitica in 1 case. The antibiogram test for the isolated Yersinia was determined. Cross-reaction between Y. pseudotuberculosis and Salmonella typhi or S. paratyphi B, and between Y. enterocolitica and Brucella was detected serologically.
Background: Pathogenic nontuberculous Mycobacteria spp. (NTM) are not known to be transmitted among persons, but may be acquired from exposure to contaminated media such as soil, food and water. We examined the spectrum of NTM isolated from human specimens in King County, WA.
Symbiodinium spp. were isolated from Porites porites (JR02F2 and RD03), Montipora capitata (JR12A7), Madracis mirabolis (RD02), Montastrea faveolata (RD04), Pocillopora damicornis (JR13E1), and an unknown coral (RD01). Growth rates and the distribution between motile gymnodinoid ...
In this study, we investigated antimicrobial resistance of Enterococcus spp. from different environmental compartments including litter from two farms, 12 surface and 28 groundwater sites in an area of intensive poultry production and litter application. The enumerated isolates (n=250) were tested ...
Lu, Jingrang; Chen, Feng; Hodson, Robert E.
The abundance of cyanophages infecting marine Synechococcus spp. increased with increasing salinity in three Georgia coastal rivers. About 80% of the cyanophage isolates were cyanomyoviruses. High cross-infectivity was found among the cyanophages infecting phycoerythrin-containing Synechococcus strains. Cyanophages in the river estuaries were diverse in terms of their morphotypes and genotypes. PMID:11425754
Dorrell, Nick; Mangan, Joseph A.; Laing, Kenneth G.; Hinds, Jason; Linton, Dennis; Al-Ghusein, Hasan; Barrell, Bart G.; Parkhill, Julian; Stoker, Neil G.; Karlyshev, Andrey V.; Butcher, Philip D.; Wren, Brendan W.
Campylobacter jejuni is the leading cause of bacterial food-borne diarrhoeal disease throughout the world, and yet is still a poorly understood pathogen. Whole genome microarray comparisons of 11 C. jejuni strains of diverse origin identified genes in up to 30 NCTC 11168 loci ranging from 0.7 to 18.7 kb that are either absent or highly divergent in these isolates. Many of these regions are associated with the biosynthesis of surface structures including flagella, lipo-oligosaccharide, and the newly identified capsule. Other strain-variable genes of known function include those responsible for iron acquisition, DNA restriction/modification, and sialylation. In fact, at least 21% of genes in the sequenced strain appear dispensable as they are absent or highly divergent in one or more of the isolates tested, thus defining 1300 C. jejuni core genes. Such core genes contribute mainly to metabolic, biosynthetic, cellular, and regulatory processes, but many virulence determinants are also conserved. Comparison of the capsule biosynthesis locus revealed conservation of all the genes in this region in strains with the same Penner serotype as strain NCTC 11168. By contrast, between 5 and 17 NCTC 11168 genes in this region are either absent or highly divergent in strains of a different serotype from the sequenced strain, providing further evidence that the capsule accounts for Penner serotype specificity. These studies reveal extensive genetic diversity among C. jejuni strains and pave the way toward identifying correlates of pathogenicity and developing improved epidemiological tools for this problematic pathogen. PMID:11591647
Wang, Liping; Jeon, Byeonghwa; Sahin, Orhan; Zhang, Qijing
Arsenic is commonly present in the natural environment and is also used as a feed additive for animal production. Poultry is a major reservoir for Campylobacter jejuni, a major food-borne human pathogen causing gastroenteritis. It has been shown that Campylobacter isolates from poultry are highly resistant to arsenic compounds, but the molecular mechanisms responsible for the resistance have not been determined, and it is unclear if the acquired arsenic resistance affects the susceptibility of Campylobacter spp. to other antimicrobials. In this study, we identified a four-gene operon that contributes to arsenic resistance in Campylobacter. This operon encodes a putative membrane permease (ArsP), a transcriptional repressor (ArsR), an arsenate reductase (ArsC), and an efflux protein (Acr3). PCR analysis of various clinical C. jejuni isolates indicated a significant association of this operon with elevated resistance to arsenite and arsenate. Gene-specific mutagenesis confirmed the role of the ars operon in conferring arsenic resistance. It was further shown that this operon is subject to regulation by ArsR, which directly binds to the ars promoter and inhibits the transcription of the operon. Arsenite inhibits the binding of ArsR to the ars promoter DNA and induces the expression of the ars genes. Mutation of the ars genes did not affect the susceptibility of C. jejuni to commonly used antibiotics. These results identify the ars operon as an important mechanism for arsenic resistance and sensing in Campylobacter.
Goldman, Cinthia G.; Matteo, Mario J.; Loureiro, Julio D.; Almuzara, Marisa; Barberis, Claudia; Vay, Carlos; Catalano, Mariana; Heredia, Sergio Rodríguez; Mantero, Paula; Boccio, Jose R.; Zubillaga, Marcela B.; Cremaschi, Graciela A.; Solnick, Jay V.; Perez-Perez, Guillermo I.; Blaser, Martin J.
The mammalian gastric and oral mucosa may be colonized by mixed Helicobacter and Campylobacter species, respectively, in individual animals. To better characterize the presence and distribution of Helicobacter and Campylobacter among marine mammals, we used PCR and 16S rDNA sequence analysis to examine gastric and oral samples from ten dolphins (Tursiops gephyreus), one killer whale (Orcinus orca), one false killer whale (Pseudorca crassidens), and three wild La Plata river dolphins (Pontoporia blainvillei). Helicobacter spp. DNA was widely distributed in gastric and oral samples from both captive and wild cetaceans. Phylogenetic analysis demonstrated two Helicobacter sequence clusters, one closely related to H. cetorum, a species isolated from dolphins and whales in North America. The second related cluster was to sequences obtained from dolphins in Australia and to gastric non-Helicobacter pylori helicobacters, and may represent a novel taxonomic group. Dental plaque sequences from four dolphins formed a third cluster within the Campylobacter genus that likely represents a novel species isolated from marine mammals. Identification of identical Helicobacter spp. DNA sequences from dental plaque, saliva and gastric fluids from the same hosts, suggests that the oral cavity may be involved in transmission. These results demonstrate that Helicobacter and Campylobacter species are commonly distributed in marine mammals, and identify taxonomic clusters that may represent novel species. PMID:21592686
Hariharan, Harry; Sharma, Shilpa; Chikweto, Alfred; Matthew, Vanessa; DeAllie, Claude
One hundred and twenty five chickens from Grenada, consisting of 77 broilers and 48 layers were examined for carriage of thermophilic campylobacters in their ceca by culture. Seventy nine percent of chickens were positive for campylobacters, with an isolation rate of 93.5% for broilers and 56.3% for layers, the difference being significant. Sixty-four pure cultures comprising 39 Campylobacter coli, 21 Campylobacter jejuni, and 4 Campyilobacter lari isolates were tested for their resistance against 7 antibiotics using the E-test. None of the isolates were resistant to chloramphenicol and gentamicin. Resistance rates to other drugs were: ampicillin, 9.4%; ciprofloxacin, 12.5%; erythromycin, 3.1%; metronidazole, 9.4%, and tetracycline, 50% with MICs of >or=256 microg/mL for tetracycline. There were no significant differences in resistance rates between C. coli and C. jejuni. Multiple resistance to >or=2 drugs was seen in 15.6% of total isolates. All C. lari isolates were resistant to ciprofloxacin, and 3 of 4 isolates had multiple drug resistance. Overall, erythromycin, which is the drug of choice for treatment of Campylobacter infections in humans, is effective in vitro against 97% of chicken isolates in Grenada.
Huang, Hongsheng; Brooks, Brian W; Lowman, Ruff; Carrillo, Catherine D
Campylobacter species, particularly thermophilic campylobacters, have emerged as a leading cause of human foodborne gastroenteritis worldwide, with Campylobacter jejuni, Campylobacter coli, and Campylobacter lari responsible for the majority of human infections. Although most cases of campylobacteriosis are self-limiting, campylobacteriosis represents a significant public health burden. Human illness caused by infection with campylobacters has been reported across Canada since the early 1970s. Many studies have shown that dietary sources, including food, particularly raw poultry and other meat products, raw milk, and contaminated water, have contributed to outbreaks of campylobacteriosis in Canada. Campylobacter spp. have also been detected in a wide range of animal and environmental sources, including water, in Canada. The purpose of this article is to review (i) the prevalence of Campylobacter spp. in animals, food, and the environment, and (ii) the relevant testing programs in Canada with a focus on the potential links between campylobacters and human health in Canada.
Zumdick, S; Petereit, F; Luftmann, H; Hensel, A
The oligomeric procyanidins (OPC) from Hawthorn leaves and flowers (Crataegi folium cum flore) are considered to be in part responsible for the cardiotonic clinical activity of the herbal material. Effective methods for rapid isolation of these heterogenous oligomeric clusters with defined molecular weight as reference compounds are not published until now. Therefore the water soluble fraction of an acetone/water (7 + 3) extract of Hawthorn leaves and flowers was fractionated by a combination of MPLC on RP-18 material and preparative HPLC using a diol stationary phase. This procedure resulted in the effective isolation of procyanidins with a distinct degree of polymerization (DP) from dimers DP2 up to tridecamers DP13. Exact mass measurements with negative ESI-TOF/MS were employed to confirm the respective structures of the isolated procyanidins.
Walochnik, Julia; Duchêne, Michael; Seifert, Karin; Obwaller, Andreas; Hottkowitz, Thomas; Wiedermann, Gerhard; Eibl, Hansjörg; Aspöck, Horst
Free-living amoebae of the genus Acanthamoeba are causing serious chronic conditions such as destructive keratitis in contact lens wearers or granulomatous amoebic encephalitis in individuals with compromised immune systems. Both are characterized by the lack of availability of sufficiently effective and uncomplicated, manageable treatments. Hexadecylphosphocholine (miltefosine) is licensed for use as a topical antineoplastic agent, but it is also active in vitro against several protozoan parasites, and it was applied very successfully for the treatment of human visceral leishmaniasis. The aim of our study was to evaluate the efficacy of hexadecylphosphocholine and other alkylphosphocholines (APCs) against Acanthamoeba spp. The in vitro activities of eight different APCs against three Acanthamoeba strains of various pathogenicities were determined. All substances showed at least amoebostatic effects, and some of them disrupted the amoebae, as shown by the release of cytoplasmic enzyme activity. Hexadecylphosphocholine exhibited the highest degree of cytotoxicity against trophozoites, resulting in complete cell death at a concentration as low as 40 μM, and also displayed significant cysticidal activity. Hexadecylphosphocholine may be a promising new candidate for the topical treatment of Acanthamoeba keratitis and, conceivably, even for the oral treatment of granulomatous amoebic encephalitis. PMID:11850250
Walochnik, Julia; Duchêne, Michael; Seifert, Karin; Obwaller, Andreas; Hottkowitz, Thomas; Wiedermann, Gerhard; Eibl, Hansjörg; Aspöck, Horst
Free-living amoebae of the genus Acanthamoeba are causing serious chronic conditions such as destructive keratitis in contact lens wearers or granulomatous amoebic encephalitis in individuals with compromised immune systems. Both are characterized by the lack of availability of sufficiently effective and uncomplicated, manageable treatments. Hexadecylphosphocholine (miltefosine) is licensed for use as a topical antineoplastic agent, but it is also active in vitro against several protozoan parasites, and it was applied very successfully for the treatment of human visceral leishmaniasis. The aim of our study was to evaluate the efficacy of hexadecylphosphocholine and other alkylphosphocholines (APCs) against Acanthamoeba spp. The in vitro activities of eight different APCs against three Acanthamoeba strains of various pathogenicities were determined. All substances showed at least amoebostatic effects, and some of them disrupted the amoebae, as shown by the release of cytoplasmic enzyme activity. Hexadecylphosphocholine exhibited the highest degree of cytotoxicity against trophozoites, resulting in complete cell death at a concentration as low as 40 microM, and also displayed significant cysticidal activity. Hexadecylphosphocholine may be a promising new candidate for the topical treatment of Acanthamoeba keratitis and, conceivably, even for the oral treatment of granulomatous amoebic encephalitis.
Loffler, Sylvia Grune; Pavan, Maria Elisa; Vanasco, Bibiana; Samartino, Luis; Suarez, Olga; Auteri, Carmelo; Romero, Graciela; Brihuega, Bibiana
Leptospirosis is the most widespread zoonosis in the world and significant efforts have been made to determine and classify pathogenic Leptospira strains. This zoonosis is maintained in nature through chronic renal infections of carrier animals, with rodents and other small mammals serving as the most important reservoirs. Additionally, domestic animals, such as livestock and dogs, are significant sources of human infection. In this study, a multiple-locus variable-number tandem repeat analysis (MLVA) was applied to genotype 22 pathogenic Leptospira strains isolated from urban and periurban rodent populations from different regions of Argentina. Three MLVA profiles were identified in strains belonging to the species Leptospira interrogans (serovars Icterohaemorrhagiae and Canicola); one profile was observed in serovar Icterohaemorrhagiae and two MLVA profiles were observed in isolates of serovars Canicola and Portlandvere. All strains belonging to Leptospira borgpetersenii serovar Castellonis exhibited the same MLVA profile. Four different genotypes were isolated from urban populations of rodents, including both mice and rats and two different genotypes were isolated from periurban populations. PMID:24676656
Loffler, Sylvia Grune; Pavan, Maria Elisa; Vanasco, Bibiana; Samartino, Luis; Suarez, Olga; Auteri, Carmelo; Romero, Graciela; Brihuega, Bibiana
Leptospirosis is the most widespread zoonosis in the world and significant efforts have been made to determine and classify pathogenic Leptospira strains. This zoonosis is maintained in nature through chronic renal infections of carrier animals, with rodents and other small mammals serving as the most important reservoirs. Additionally, domestic animals, such as livestock and dogs, are significant sources of human infection. In this study, a multiple-locus variable-number tandem repeat analysis (MLVA) was applied to genotype 22 pathogenic Leptospira strains isolated from urban and periurban rodent populations from different regions of Argentina. Three MLVA profiles were identified in strains belonging to the species Leptospira interrogans (serovars Icterohaemorrhagiae and Canicola); one profile was observed in serovar Icterohaemorrhagiae and two MLVA profiles were observed in isolates of serovars Canicola and Portlandvere. All strains belonging to Leptospira borgpetersenii serovar Castellonis exhibited the same MLVA profile. Four different genotypes were isolated from urban populations of rodents, including both mice and rats and two different genotypes were isolated from periurban populations.
Mohan, Vathsala; Stevenson, Mark; Marshall, Jonathan; Fearnhead, Paul; Holland, Barbara R; Hotter, Grant; French, Nigel P
Abstract A repeated cross-sectional study was conducted to determine the prevalence of Campylobacter spp. and the population structure of C. jejuni in European starlings and ducks cohabiting multiple public access sites in an urban area of New Zealand. The country's geographical isolation and relatively recent history of introduction of wild bird species, including the European starling and mallard duck, create an ideal setting to explore the impact of geographical separation on the population biology of C. jejuni, as well as potential public health implications. A total of 716 starling and 720 duck fecal samples were collected and screened for C. jejuni over a 12 month period. This study combined molecular genotyping, population genetics and epidemiological modeling and revealed: (i) higher Campylobacter spp. isolation in starlings (46%) compared with ducks (30%), but similar isolation of C. jejuni in ducks (23%) and starlings (21%), (ii) significant associations between the isolation of Campylobacter spp. and host species, sampling location and time of year using logistic regression, (iii) evidence of population differentiation, as indicated by FST, and host-genotype association with clonal complexes CC ST-177 and CC ST-682 associated with starlings, and clonal complexes CC ST-1034, CC ST-692, and CC ST-1332 associated with ducks, and (iv) greater genetic diversity and genotype richness in ducks compared with starlings. These findings provide evidence that host-associated genotypes, such as the starling-associated ST-177 and ST-682, represent lineages that were introduced with the host species in the 19th century. The isolation of sequence types associated with human disease in New Zealand indicate that wild ducks and starlings need to be considered as a potential public health risk, particularly in urban areas. We applied molecular epidemiology and population genetics to obtain insights in to the population structure, host-species relationships, gene flow and
Prabhakaran, Pranesha; Ashraf, Muhammad Aqeel; Aqma, Wan Syaidatul
Increased anthropogenic activities such as mining have contributed to accumulation of heavy metals in the environment. Microorganisms that are found in the contaminated area develop resistance toward the heavy metals after a prolonged exposure. Screenings for bacterial resistance to metal tin were conducted using isolated Thiobacillus spp. from a tin mining area at Bestari Jaya, Selangor. Two isolated Thiobacillus spp. able to show growth without inhibition up to 300 ppm. IC50 that indicate 50% inhibition of the bacterial growth for both isolates, TB1 and TB2 are 496.03 ppm and 514.27 ppm respectively under tin-stressed condition. Adaptation of these microorganisms toward heavy metal could be exploited for bioremediation of heavy metals at contaminated sites.
Shirzad Aski, Hesamaddin; Tabatabaei, Mohammad; Khoshbakht, Rahem; Raeisi, Mojtaba
This study is conducted to determine the occurrence and antimicrobial resistance of Arcobacter spp. isolated from clinically healthy food animals. A total of 308 samples from cattle (200) and sheep (108) were collected from Shiraz slaughterhouse, southern Iran to investigate the presence of the important Arcobacter spp. using cultivation and Polymerase Chain Reaction (PCR) methods. Antimicrobial susceptibility of Arcobacter isolates was determined for 18 antibiotics using disk diffusion method. Among 308 samples, 27 (8.7%) and 44 (14.28%) were positive for the presence of Arcobacter species with cultivation and PCR procedures, respectively. The predominant species was A. butzleri in both cattle (58.33%) and sheep (55%). In addition, concurrent incidence of the species was observed in 25% of the positive samples. All Arcobacter isolates were resistant to rifampicin, vancomycin, ceftriaxone, trimethoprim and cephalothin. The isolates showed high susceptibility to tetracycline, oxytetracycline, erythromycin, ciprofloxacin, kanamycin, amikacin, gentamicin and enrofloxacin. No significant difference among cattle and sheep isolates in resistance pattern was observed. The results indicate that cattle and sheep are significant intestinal carriers for Arcobacter spp. Moreover, tetracycline and aminoglycosides showed great effects on Arcobacter species in antibiogram test and can be used for treatment of human Arcobacter infections.
Beeton, M L; Spiller, O B
There is growing global concern regarding the rise of antibiotic-resistant organisms. Many of these reports have focused on various Gram-positive and Gram-negative pathogens, with little attention to the genus Ureaplasma. Ureaplasma spp. are associated with numerous infectious diseases affecting pregnant women, neonates and the immunocompromised. Treatment options are extremely limited due to high levels of intrinsic resistance resulting from the unique physiology of these organisms and further restricted in cases of the developing fetus or neonate, often limiting therapeutic options to predominantly macrolides or rarely fluoroquinolones. The increasing presence of macrolide- and fluoroquinolone-resistant strains among neonatal infections may result in pan-drug resistance and potentially untreatable conditions. Here, we review the requirements for accurate measurement of antimicrobial susceptibility, provide a comprehensive review of the antimicrobial resistance (AMR) for Ureaplasma species in the literature and contextualize these results relative to some investigators' reliance on commercial kits that are not CLSI compliant when determining AMR. The dramatic variation in the resistance patterns and impact of high levels of AMR amongst neonatal populations suggests the need for continued surveillance. Commercial kits represent an excellent tool for initial antibiotic susceptibility determination and screening. However, AMR reporting must utilize internationally standardized methods, as high-titre samples, or Mycoplasma hominis-contaminated samples routinely give false AMR results. Furthermore, there is a requirement for future reports to determine the underlying AMR mechanisms and determine whether expanding AMR is due to spontaneous mutation, transmission of resistance genes on mobile elements or selection and expansion of resistant clones.
Eryılmaz, Müjde; Akın, Ahmet; Arıkan Akan, Ozay
Nosocomial infections which exhibit an increasing trend worldwide, are important contributors to morbidity and mortality. Most bacteria that cause nosocomial infections can retain their viability even after exposure to disinfectants in routine practice. This study was conducted to determine the susceptibilities of nosocomial Staphylococcus aureus and Enterococcus spp. isolates to various disinfectants. A total of 30 S.aureus [16 were methicillin-resistant (MRSA), 14 were methicillin-susceptible (MSSA)] and 21 Enterococcus spp. (13 E.faecalis, 7 E.faecium, 1 non-typable Enterococcus spp.) strains isolated from clinical samples of hospitalized patients as nosocomial infection agents in the Central Microbiology Laboratory of Ibn-i Sina Hospital, Ankara University, Faculty of Medicine, were included in the study. Glutaraldehyde (2% wt/vol), chlorhexidine gluconate (4% wt/vol), 2-propanol (70% vol/vol), povidone iodine (7.5% wt/vol), povidone iodine (10% wt/vol) and hydrogen peroxide (3% wt/vol) susceptibilities of the isolates were investigated by quantitative suspension test at contact times of 3, 5, and 10 minutes. All of the isolates were found susceptible to glutaraldehyde (2%), chlorhexidine gluconate (4%), povidone iodine (7.5%), povidone iodine (10%) and 2-propanol (70%) at all tested contact times. However, 12 S.aureus (5 MSSA, 7 MRSA) and 3 enterococci (2 E.faecium, 1 E.faecalis) isolates were found susceptible to hydrogen peroxide (3%) at 3 minutes contact time; 11 S.aureus (4 MSSA, 7 MRSA) and 7 E.faecalis isolates were found susceptible at 5 minutes contact time, and 6 S.aureus (4 MSSA, 2 MRSA) and 3 enterococci (1 E.faecium, 2 E.faecalis) isolates were found susceptible at 10 minutes contact time. One MSSA and 8 enterococci (4 E.faecium, 3 E.faecalis, 1 Enterococcus spp.) isolates were found resistant to hydrogen peroxide (3%) at 10 minutes contact time. In conclusion, glutaraldehyde (2%), chlorhexidine gluconate (4%), povidone iodine (7.5%), povidone
Michaud, Sophie; Ménard, Suzanne; Arbeit, Robert D
The goal of the present study was to assess the contribution of real-time molecular typing, used alone or with clinical surveillance, to the prompt identification of clusters of Campylobacter enteritis. Potential poultry sources were sought by comparing the pulsed-field gel electrophoresis genotypes of human and fresh whole retail chicken isolates collected during the same study period. Among 183 human isolates, 82 (45%) had unique genotypes, 72 (39%) represented 26 clusters of 2 to 7 isolates each, and 29 (16%) represented three clusters of 8 to 11 isolates each. Molecular typing was useful for the confirmation of outbreaks suspected on the basis of epidemiological surveillance, but for most small clusters, no epidemiological link could be established. Thus, the added value of real-time molecular typing is questionable, since the numerous small clusters identified were of unclear public health significance. Among 177 chickens, 41 (23%) yielded campylobacter isolates; of these, 19 (46%) had genotypes similar to those of 41 (22%) human isolates. However, a temporal association was demonstrated in only a minority of cases, and most genotypes were present only in a single species, suggesting that sources other than chickens are important in human campylobacteriosis. Further investigation with samples from water and other possible environmental sources is needed to define the most efficient strategy for the application of molecular typing and identification of the source(s) of sporadic cases of campylobacteriosis.
Kittinger, Clemens; Lipp, Michaela; Baumert, Rita; Folli, Bettina; Koraimann, Günther; Toplitsch, Daniela; Liebmann, Astrid; Grisold, Andrea J.; Farnleitner, Andreas H.; Kirschner, Alexander; Zarfel, Gernot
Spread and persistence of antibiotic resistance pose a severe threat to human health, yet there is still lack of knowledge about reservoirs of antibiotic resistant bacteria in the environment. We took the opportunity of the Joint Danube Survey 3 (JDS3), the world's biggest river research expedition of its kind in 2013, to analyse samples originating from different sampling points along the whole length of the river. Due to its high clinical relevance, we concentrated on the characterization of Pseudomonas spp. and evaluated the resistance profiles of Pseudomonas spp. which were isolated from eight sampling points. In total, 520 Pseudomonas isolates were found, 344 (66.0%) isolates were identified as Pseudomonas putida, and 141 (27.1%) as Pseudomonas fluorescens, all other Pseudomonas species were represented by less than five isolates, among those two P. aeruginosa isolates. Thirty seven percent (37%) of all isolated Pseudomonas species showed resistance to at least one out of 10 tested antibiotics. The most common resistance was against meropenem (30.4%/158 isolates) piperacillin/tazobactam (10.6%/55 isolates) and ceftazidime (4.2%/22 isolates). 16 isolates (3.1%/16 isolates) were multi-resistant. For each tested antibiotic at least one resistant isolate could be detected. Sampling points from the upper stretch of the River Danube showed more resistant isolates than downriver. Our results suggest that antibiotic resistance can be acquired by and persists even in Pseudomonas species that are normally not in direct contact with humans. A possible scenario is that these bacteria provide a reservoir of antibiotic resistance genes that can spread to related human pathogens by horizontal gene transfer. PMID:27199920
Jacobs, Liezl; Chenia, Hafizah Y
An increasing incidence of multidrug resistance amongst Aeromonas spp. isolates, which are both fish pathogens and emerging opportunistic human pathogens, has been observed worldwide. This can be attributed to the horizontal transfer of mobile genetic elements, viz.: plasmids and class 1 integrons. The antimicrobial susceptibilities of 37 Aeromonas spp. isolates, from tilapia, trout and koi aquaculture systems, were determined by disc-diffusion testing. The plasmid content of each isolate was examined using the alkaline lysis protocol. Tet determinant type was determined by amplification using two degenerate primer sets and subsequent HaeIII restriction. The presence of integrons was determined by PCR amplification of three integrase genes, as well as gene cassettes, and the qacEDelta1-sulI region. Thirty-seven Aeromonas spp. isolates were differentiated into six species by aroA PCR-RFLP, i.e., A. veronii biovar sobria, A. hydrophila, A. encheleia, A. ichtiosoma, A. salmonicida, and A. media. High levels of resistance to tetracycline (78.3%), amoxicillin (89.2%), and augmentin (86.5%) were observed. Decreased susceptibility to erythromycin was observed for 67.6% of isolates. Although 45.9% of isolates displayed nalidixic acid resistance, majority of isolates were susceptible to the fluoroquinolones. The MAR index ranged from 0.12 to 0.59, with majority of isolates indicating high-risk contamination originating from humans or animals where antibiotics are often used. Plasmids were detected in 21 isolates, with 14 of the isolates displaying multiple plasmid profiles. Single and multiple class A family Tet determinants were observed in 27% and 48.7% of isolates, respectively, with Tet A being the most prevalent Tet determinant type. Class 1 integron and related structures were amplified and carried different combinations of the antibiotic resistance gene cassettes ant(3'')Ia, aac(6')Ia, dhfr1, oxa2a and/or pse1. Class 2 integrons were also amplified, but the
Er, Buket; Demirhan, Burak; Onurdag, Fatma Kaynak; Ozgacar, Selda Özgen; Oktem, Aysel Bayhan
Salmonella spp. are widespread foodborne pathogens that contaminate egg and poultry meats. Attachment, colonization, as well as biofilm formation capacity of Salmonella spp. on food and contact surfaces of food may cause continuous contamination. Biofilm may play a crucial role in the survival of salmonellae under unfavorable environmental conditions, such as in animal slaughterhouses and processing plants. This could serve as a reservoir compromising food safety and human health. Addition of antimicrobial preservatives extends shelf lives of food products, but even when products are supplemented with adequate amounts of preservatives, it is not always possible to inhibit the microorganisms in a biofilm community. In this study, our aims were i) to determine the minimum inhibitory concentrations (MIC) and minimum biofilm inhibitory concentrations (MBIC) of selected preservatives against planktonic and biofilm forms of Salmonella spp. isolated from chicken samples and Salmonella Typhimurium SL1344 standard strain, ii) to show the differences in the susceptibility patterns of same strains versus the planktonic and biofilm forms to the same preservative agent, and iii) to determine and compare antimicrobial and antibiofilm effects of selected food preservatives against Salmonella spp. For this purpose, Salmonella Typhimurium SL1344 standard strain and 4 Salmonella spp. strains isolated from chicken samples were used. Investigation of antimicrobial and antibiofilm effects of selected food preservatives against Salmonella spp. was done according to Clinical and Laboratory Standards Institute M100-S18 guidelines and BioTimer assay, respectively. As preservative agents, pure ciprofloxacin, sodium nitrite, potassium sorbate, sodium benzoate, methyl paraben, and propyl paraben were selected. As a result, it was determined that MBIC values are greater than the MIC values of the preservatives. This result verified the resistance seen in a biofilm community to food
Tiewcharoen, Supathra; Komalamisra, Narumon; Junnu, Virach
A genetic approach was cited for species detection of the ameba genus Naegleria using allozyme electrophoresis to characterize the trophozoite stage of three strains of Naegleria fowleri isolated from patients with primary amebic meningoencephalitis, five thermophilic (45 degrees C) Naegleria spp isolated from natural water sources in the Taling Chan district, and a reference control strain, Naegleria fowleri CDC VO 3081. Isoenzymes of ameba whole-cell extracts were analyzed by vertical polyacrylamide slab gel electrophoresis to determine whether there was any correlation between different strains of the ameba. The results showed that five out of fifteen enzymes; aldehyde oxidase (ALDOX), aldolase (ALD), a-glycerophosphate dehydrogenase (a-GPDH), xanthine dehydrogenase (XDH), and glutamate oxaloacetate transaminase (GOT), were undetectable in the pathogenic strains, while the other enzymes; esterase (EST), fumerase (FUM), glucose-6-phosphate dehydrogenase (G-6-PDH), glucose phosphate isomerase (GPI), isocitate dehydrogenase (IDH), lactate dehydrogenase (LDH), leucine aminopeptidase (LAP), malic enzyme (ME), glucose phosphomutase (GPM), and malate dehydrogenase (MDH), were detected. Naegleria fowleri strains were biochemically the most homogeneous. They showed intraspecific isoenzyme variation that allowed them to be grouped. In contrast, the allozyme patterns (EST 1-7, IDH) of Naegleria spp isolated from the environment showed interspecific isoenzyme variations from the pathogenic Naegleria strain. In conclusion, this study recognized the zymograms of the Naegleria fowleri strains were heterogenically different from the thermophilic 45 degrees C Naegleria spp isolated from the environment.
Chen, Wanyi; Yang, Jielin; You, Chunping; Liu, Zhenmin
Cronobacter spp. has caused life-threatening neonatal infections mainly resulted from consumption of contaminated powdered infant formula. A total of 102 vegetable samples from retail markets were evaluated for the presence of Cronobacter spp. Thirty-five presumptive Cronobacter isolates were isolated and identified using API 20E and 16S rDNA sequencing analyses. All isolates and type strains were characterized using enterobacterial repetitive intergenic consensus sequence PCR (ERIC-PCR), and genetic profiles of cluster analysis from this molecular typing test clearly showed that there were differences among isolates from different vegetables. A polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) based on the amplification of the gyrB gene (1258 bp) was developed to differentiate among Cronobacter species. A new PCR-RFLP assay based on the amplification of the gyrB gene using Alu I and Hinf I endonuclease combination is established and it has been confirmed an accurate and rapid subtyping method to differentiate Cronobacter species. Sequence analysis of the gyrB gene was proven to be suitable for the phylogenetic analysis of the Cronobacter strains, which has much better resolution based on SNPs in the identification of Cronobacter species specificity than PCR-RFLP and ERIC-PCR. Our study further confirmed that vegetables are one of the most common habitats or sources of Cronobacter spp. contamination in the middle-east coastline of China.
Mohammed, Hussni O; Stipetic, Korana; Salem, Ahmed; McDonough, Patrick; Chang, Yung Fu; Sultan, Ali
Escherichia coli O157:H7, non-O157 E. coli, and Campylobacter spp. are among the top-ranked pathogens that threaten the safety of food supply systems around the world. The associated risks and predisposing factors were investigated in a dynamic animal population using a repeat-cross-sectional study design. Animal and environmental samples were collected from dairy and camel farms, chicken processing plants, and abattoirs and analyzed for the presence of these pathogens using a combination of bacterial enrichment and real-time PCR tests without culture confirmation. Data on putative risk factors were also collected and analyzed. E. coli O157:H7 was detected by PCR at higher levels in sheep and camel feces than in cattle feces (odds ratios [OR], 6.8 and 21.1, respectively). Although the genes indicating E. coli O157:H7 were detected at a relatively higher rate (4.3%) in fecal samples from dairy cattle, they were less common in milk and udder swabs from the same animals (1 and 2%, respectively). Among the food adulterants, E. coli O103 was more common in cattle fecal samples, whereas O26 was more common in sheep feces and O45 in camel feces compared with cattle (OR, 2.6 and 3.1, respectively). The occurrence of E. coli in the targeted populations differed by the type of sample and season of the year. Campylobacter jejuni and Campylobacter coli were more common in sheep and camel feces than in cattle feces. Most of the survey and surveillance of E. coli focused on serogroup O157 as a potential foodborne hazard; however, based on the PCR results, non-O157 Shiga toxin-producing E. coli serotypes appeared to be more common, and efforts should be made to include them in food safety programs.
Choo, L C; Saleha, A A; Wai, S S; Fauziah, N
Insects, in particular house flies and cockroaches, have been shown to be associated with the spread of pathogens in livestock farms and in human disease outbreaks: among these pathogens are salmonellae and campylobacters. A total of 60 flies were caught in three locations: an animal teaching facility and a cafeteria in a university campus, and a poultry farm. Five percent (5%) and 13.3% of flies sampled were found to carry Campylobacter and Salmonella, respectively.
Paszko-Kolva, C; Yamamoto, H; Shahamat, M; Sawyer, T K; Morris, G; Colwell, R R
Forty eyewash units were sampled for protozoa, bacteria, and fungi. Total heterotrophic bacterial counts on nutrient agar and R2A agar (Difco Laboratories, Detroit, Mich.) ranged from 0 to 10(5) CFU/ml, with Pseudomonas spp. being the most frequently isolated. Total counts of 10(4) and 10(8) cells per ml were obtained with the acridine orange staining procedure. All samples were examined for Legionella spp. by direct fluorescent-antibody staining and by culturing on buffered charcoal-yeast extract agar containing alpha-ketoglutarate and glycine and supplemented with cycloheximide, vancomycin, and polymyxin B. DNA-DNA hybridization was used to confirm identification of the Legionella isolates. Legionellae were detected in 35 of 40 (87.5%) samples by direct fluorescent-antibody staining, with 3 samples yielding both Legionella spp. and amoebae. Amoebae identified as Hartmannella, Vahlkampfia, Acanthamoeba, and Cochliopodium spp. were detected in 19 of 40 (47:5%) samples. Sabouraud dextrose agar was used to obtain a crude estimate of viable fungal populations, pH, hardness, and ammonia, alkalinity, chlorine, copper, and iron contents were recorded for all water samples collected from eyewash stations; 33% of the samples had greater than or equal to 10 mg of CO2 per liter. It is concluded that eyewash stations not regularly flushed and/or cleaned and used to flush traumatized eye tissue may be a source of infection and can contaminate laboratory environments via aerosol transmission. PMID:2036003
Chaillon, Antoine; Baty, Gaelle; Lauvin, Marie Agnès; Besnier, Jean Marc; Goudeau, Alain; Lanotte, Philippe
Campylobacter spp. are common causes of gastrointestinal infections. Campylobacter fetus is a much rarer pathogen in humans, and usually causes bacteraemia and systemic complications in patients with predisposing conditions. We report a case of spondylodiscitis caused by C. fetus subsp. fetus as revealed by vertebral biopsy culture. This identification was confirmed by sequencing the 16S rRNA gene and by phylogenetic analysis. Treatment consisted of 6 weeks antimicrobial therapy combined with a strict initial immobilization, followed by a re-education program. The patient's recovery was uneventful.
Ghorbanalizadgan, Mahdi; Bakhshi, Bita; Najar-Peerayeh, Shahin
The aims of this study were to determine the genetic relatedness among 20 clinical Campylobacter jejuni samples isolated from children with diarrhea in Iran and to introduce the best method of discrimination based on flagellin gene (flaA) sequence divergence. A total of 400 stool specimens were obtained from children under 5 years of age from July 2012 to June 2013. Primers were designed based on conserved sequences flanking the flaA gene that encompassed and amplified the entire flaA gene and followed by sequencing and data analysis with MEGA version 6.0.6 software. Ninety amino acids and 560 nucleotide polymorphic sequences were detected within 1,681 bp of the flaA sequence of which 43 (2.5%) and 12 (0.7%) were singletons, respectively. New repeat boxes within the flaA sequences were found in this study. Unweighted Pair Group Method with Arithmetic Mean dendrogram based on nucleotides of the full length flaA gene, the flaA short variable region gene and the in silico flaA phylogenic tree of DdeI restriction fragment length polymorphism (RFLP) profiles produced very similar clustering with a diversity index of 0.86 for each of the 3 methods. We conclude that flaA typing based on DdeI RFLP of the PCR products is a cheap, rapid, and reliable method for the epidemiological study of C. jejuni isolates of clinical origin in resource-limited regions or in large-scale population surveillance.
Octavia, Sophie; Day, Andrew S.; Riordan, Stephen M.; Grimm, Michael C.; Lan, Ruiting; Lemberg, Daniel; Tran, Thi Anh Tuyet; Zhang, Li
Background Campylobacter concisus, a bacterium colonizing the human oral cavity, has been shown to be associated with inflammatory bowel disease (IBD). This study investigated if patients with IBD are colonized with specific oral C. concisus strains that have potential to cause enteric diseases. Methodology Seventy oral and enteric C. concisus isolates obtained from eight patients with IBD and six controls were examined for housekeeping genes by multilocus sequence typing (MLST), Caco2 cell invasion by gentamicin-protection-assay, protein analysis by mass spectrometry and SDS-PAGE, and morphology by scanning electron microscopy. The whole genome sequenced C. concisus strain 13826 which was isolated from an individual with bloody diarrhea was included in MLST analysis. Principal Findings MLST analysis showed that 87.5% of individuals whose C. concisus belonged to Cluster I had inflammatory enteric diseases (six IBD and one with bloody diarrhea), which was significantly higher than that in the remaining individuals (28.6%) (P<0.05). Enteric invasive C. concisus (EICC) oral strain was detected in 50% of patients with IBD and none of the controls. All EICC strains were in Cluster 1. The C. concisus strain colonizing intestinal tissues of patient No. 1 was closely related to the oral C. concisus strain from patient No. 6 and had gene recombination with the patient’s own oral C. concisus. The oral and intestinal C. concisus strains of patient No. 3 were the same strain. Some individuals were colonized with multiple oral C. concisus strains that have undergone natural recombination. Conclusions This study provides the first evidence that patients with IBD are colonized with specific oral C. concisus strains, with some being EICC strains. C. concisus colonizing intestinal tissues of patients with IBD at least in some instances results from an endogenous colonization of the patient’s oral C. concisus and that C. concisus strains undergo natural recombination. PMID
Klena, John D.; Parker, Craig T.; Knibb, Krista; Ibbitt, J. Claire; Devane, Phillippa M. L.; Horn, Sharon T.; Miller, William G.; Konkel, Michael E.
We describe a multiplex PCR assay to identify and discriminate between isolates of Campylobacter coli, Campylobacter jejuni, Campylobacter lari, and Campylobacter upsaliensis. The C. jejuni isolate F38011 lpxA gene, encoding a UDP-N-acetylglucosamine acyltransferase, was identified by sequence analysis of an expression plasmid that restored wild-type lipopolysaccharide levels in Escherichia coli strain SM105 [lpxA(Ts)]. With oligonucleotide primers developed to the C. jejuni lpxA gene, nearly full-length lpxA amplicons were amplified from an additional 11 isolates of C. jejuni, 20 isolates of C. coli, 16 isolates of C. lari, and five isolates of C. upsaliensis. The nucleotide sequence of each amplicon was determined, and sequence alignment revealed a high level of species discrimination. Oligonucleotide primers were constructed to exploit species differences, and a multiplex PCR assay was developed to positively identify isolates of C. coli, C. jejuni, C. lari, and C. upsaliensis. We characterized an additional set of 41 thermotolerant isolates by partial nucleotide sequence analysis to further demonstrate the uniqueness of each species-specific region. The multiplex PCR assay was validated with 105 genetically defined isolates of C. coli, C. jejuni, C. lari, and C. upsaliensis, 34 strains representing 12 additional Campylobacter species, and 24 strains representing 19 non-Campylobacter species. Application of the multiplex PCR method to whole-cell lysates obtained from 108 clinical and environmental thermotolerant Campylobacter isolates resulted in 100% correlation with biochemical typing methods. PMID:15583280
Ghosh, Santanu; Pazhani, Gururaja P; Niyogi, Swapan Kumar; Nataro, James P; Ramamurthy, Thandavarayan
Phenotypic and genetic characteristics of Shigella spp. isolated from diarrhoeal and asymptomatic children aged up to 5 years were analysed in this study. In total, 91 and 17 isolates were identified from diarrhoeal (case) and asymptomatic (control) children, respectively. All the isolates were tested for antimicrobial resistance, the presence of integrons, plasmid-mediated quinolone resistance (PMQR), virulence-associated genes and Shigella pathogenicity island (SH-PAI). The majority of the Shigella spp. from cases (68.1%) and controls (82.3%) were found to be resistant to fluoroquinolones. Integron carriage was detected more in cases (76.9%) than in controls (35.5%). Atypical class 1 integron was detected exclusively in Shigella flexneri from cases but not from the controls. PMQR genes such as aac(6')-Ib-cr and qnrS1 were detected in 82.4 and 14.3% of the isolates from cases and in 53 and 17.6% in controls, respectively. Shigella isolates from cases as well as from controls were positive for the invasive plasmid antigen H-encoding gene ipaH. The other virulence genes such as virF, sat, setA, setB, sen and ial were detected in Shigella isolates in 80.2, 49.4, 27.4, 27.4, 80.2 and 79.1% of cases and in 64.7, 52.9, 17.6, 17.6, 64.7 and 64.7% of controls, respectively. The entire SH-PAI was detected in S. flexneri serotype 2a from cases and controls. In an isolate from a control child, the SH-PAI was truncated. Integrons, PMQR and virulence-encoding genes were detected more frequently in cases than in controls. In diarrhoea endemic areas, asymptomatic carriers may play a crucial role in the transmission of multidrug-resistant Shigella spp. with all the putative virulence genes.
The annual Sandhill crane (Grus canadensis) migration through Nebraska is thought to be a major source of fecal pollution to the Platte River, but of unknown human health risk. To better understand potential risks, the presence of Campylobacter species and fecal bacteria were exa...
A total of 689 co-grazing small ruminants along with 446 wild-living birds were tested during two springs and autumns under two management systems at two Mid-Atlantic locations (~187 km in aerial distance) of the U.S. Fecal shedding of Campylobacter jejuni and Salmonella were, respectively, detected...
significant high isolation rate of C. jejuni in Thai- land and potential role of the FspA protein as one of vaccine candidate lead us to further...foreigners and Thai adults at Bumrungrad Hospital in Bangkok. All C. jejuni isolates were kept at -70 oc in glycerol medium and later were
Jones, D R; Guard, J; Gast, R K; Buhr, R J; Fedorka-Cray, P J; Abdo, Z; Plumblee, J R; Bourassa, D V; Cox, N A; Rigsby, L L; Robison, C I; Regmi, P; Karcher, D M
The housing of laying hens is important for social, industrial, and regulatory aspects. Many studies have compared hen housing systems on the research farm, but few have fully examined commercial housing systems and management strategies. The current study compared hens housed in commercial cage-free aviary, conventional cage, and enriched colony cage systems. Environmental and eggshell pool samples were collected from selected cages/segments of the housing systems throughout the production cycle and monitored for Salmonella and Campylobacter prevalence. At 77 wk of age, 120 hens per housing system were examined for Salmonella and Campylobacter colonization in the: adrenal glands, spleen, ceca, follicles, and upper reproductive tract. All isolates detected from environmental swabs, eggshell pools, and tissues were identified for serotype. Two predominant Salmonella were detected in all samples:S.Braenderup andS.Kentucky.Campylobacter coli and C. jejuni were the only Campylobacter detected in the flocks. Across all housing systems, approximately 7% of hens were colonized with Salmonella, whereas >90% were colonized with Campylobacter Salmonella Braenderup was the isolate most frequently detected in environmental swabs (P<0.0001) and housing system impacted Salmonella spp. shedding (P<0.0001).Campylobacter jejuni was the isolate most frequently found in environmental swabs (P<0.01), while housing system impacted the prevalence of C. coli and jejuniin ceca (P<0.0001). The results of this study provide a greater understanding of the impact of hen housing systems on hen health and product safety. Additionally, producers and academia can utilize the findings to make informed decisions on hen housing and management strategies to enhance hen health and food safety.
Horvath, Lynn L.; Hospenthal, Duane R.; Murray, Clinton K.; Dooley, David P.
CHROMagar Candida is a selective and differential chromogenic medium that has been shown to be useful for identification of Candida albicans, Candida krusei, Candida tropicalis, and perhaps Candida glabrata. Colony morphology and color have been well defined when CHROMagar Candida has been used to isolate yeast directly from clinical specimens, including stool, urine, respiratory, vaginal, oropharyngeal, and esophageal sources. Direct isolation of yeast on CHROMagar Candida from blood cultures has not been evaluated. We evaluated whether the color and colony characteristics produced by Candida spp. on CHROMagar Candida were altered when yeasts were isolated directly from blood cultures. Fifty clinical isolates of Candida were inoculated into aerobic and anaerobic blood culture bottles and incubated at 35°C in an automated blood culture system. When growth was detected, an aliquot was removed and plated onto CHROMagar Candida. As a control, CHROMagar Candida plates were inoculated with the same isolate of yeast grown on Sabouraud dextrose agar simultaneously. No significant difference was detected in color or colony morphology between the blood and control isolates in any of the tested organisms. All C. albicans (n = 12), C. tropicalis (n = 12), C. glabrata (n = 9), and C. krusei (n = 5) isolates exhibited the expected species-specific colony characteristics and color, whether isolated directly from blood or from control cultures. CHROMagar Candida can be reliably used for direct isolation of yeast from blood cultures. Direct isolation could allow mycology laboratories to more rapidly identify Candida spp., enable clinicians to more quickly make antifungal agent selections, and potentially decrease patient morbidity and mortality. PMID:12791890
Marcos-Zambrano, Laura Judith; Escribano, Pilar; Sánchez-Carrillo, Carlos; Bouza, Emilio; Guinea, Jesús
Trailing is a well-known phenomenon that is defined as reduced but persistent visible growth of Candida spp. at fluconazole concentrations above the MIC. Trailing is commonly detected using the CLSI M27-A3 method, although little is known about its frequency when investigated with EUCAST. We assessed the frequency and scope of fluconazole trailing after using EUCAST EDef 7.2. against a large number of Candida spp. isolates from patients with candidemia. We studied 639 fluconazole-susceptible non-krusei Candida spp. isolates from 570 patients admitted to Gregorio Marañón Hospital. Isolates were tested in vitro for fluconazole susceptibility according to the EUCAST EDef 7.2 procedure; trailing was defined as the presence of any residual growth in wells containing fluconazole concentrations above the MIC. According to the mean percentage of trailing observed, isolates were classified as residual trailers (0.1-5%), slight trailers (6%-10%), moderate trailers (11%-15%), and heavy trailers (>15%). The relationship between trailing and genotyping was assessed. The mean overall percentage of trailing was 6.8%, with C. albicans and C. tropicalis showing the highest percentages (9.75% and 9.29%, respectively; P < .001). C. albicans and C. tropicalis had the highest percentage of heavy trailers (>15%). Trailing was not genotype-specific. Fluconazole trailing was observed frequently when EUCAST was used for antifungal susceptibility testing, particularly in isolates of C. albicans and C. tropicalis The cut-off proposed enabled us to classify the isolates according to the degree of trailing and can be used as the basis for future studies to evaluate the clinical impact of this phenomenon.
Karunasagar, Anusha; Maiti, Biswajit; Shekar, Malathi; Shenoy M, Shalini; Karunasagar, Indrani
The prevalence of OXA-type carbapenemase genes, ISAba1 insertion sequence, carbapenem resistance, biofilm forming ability and genetic heterogeneity in clinical isolates of Acinetobacter spp. from hospitals in Mangalore, South India was studied. Based on the presence of the bla(OXA-51) -like gene, the 62 isolates of Acinetobacter spp. were identified as 48 A. baumannii and 14 other Acinetobacter spp. The prevalence of bla(OXA-23) -like, bla(OXA-24) -like and bla(OXA-58) -like genes in A. baumannii was 47.9%, 22.9% and 4.2%, while in other Acinetobacter spp. it was 28.5%, 64.3% and 35.7% respectively. Several A. baumannii isolates (16/48) harbored the insertion sequence ISAba1 in the upstream region of the bla(OXA-23) -like gene. Resistance to meropenem was seen in 39.6% and 14.2% of A. baumannii and other Acinetobacter spp. isolates, respectively. The ability to form biofilm was observed to be higher among A. baumannii in comparison to other Acinetobacter spp. The present study shows that bla(OXA-23) -like genes are more common in A. baumannii,whereas bla(OXA-24) -like genes are common to other Acinetobacter spp. The study revealed genetic heterogeneity among the isolates, indicating multiple sources in the hospitals.
Soltan Dallal, Mohammad Mehdi; Mazaheri Nezhad Fard, Ramin; Kavan Talkhabi, Morteza; Aghaiyan, Leyla; Salehipour, Zohre
Background Aeromonas spp. cause various intestinal and extraintestinal diseases. These bacteria are usually isolated from fecal samples, especially in children under five years old. The aim of this study was to assess the prevalence of Aeromonas spp. and their antimicrobial resistance profile in children with diarrhea referred to the Children Medical Center in Tehran, between 2013 and 2014. Methods A total number of 391 stool samples were collected from children with ages between 1 day and 14 years old, with diarrhea (acute or chronic), referred to the Children Hospital, Tehran, Iran, between 2013 and 2014. Samples were enriched in alkaline peptone water broth for 24 hours at 37 °C and then cultured. Suspicious colonies were analyzed through biochemical tests. Furthermore, antimicrobial susceptibility tests were carried out for the isolates. Isolates were further studied for act, ast, alt, aerA and hlyA virulence genes using polymerase chain reaction. Results In total, 12 isolates (3.1%) were identified as Aeromonas spp.; all were confirmed using the API-20E test. Of these isolates, five A. caviae (42%), four A. veronii (33%) and three A. hydrophila (25%) were identified in cases with gastroenteritis. Second to ampicillin (which was included in the growth medium used), the highest rate of antimicrobial resistance was seen against nalidixic acid and trimethoprim-sulfamethoxazole (5 isolates each, 41.6%) and the lowest rate of antimicrobial resistance was seen against gentamicin, amikacin and cefepime (none of the isolates). Results included 76.4% act, 64.7% ast, 71.5% alt, 83.3% aerA and 11.7% hlyA genes. Conclusion Aeromonas spp. are important due to their role in diarrhea in children; therefore, isolation and identification of these fecal pathogens should seriously be considered in medical laboratories. Since virulence genes play a significant role in gastroenteritis symptoms caused by these bacteria, Aeromonas species that include virulence genes are potentially
Fernández, Heriberto; Hitschfeld, Marianne
The prevalence of Campylobacter jejuni and Campylobacter coli and their biotypes in beef and dairy cattle from the South of Chile was established. Campylobacter were statistically more prevalent among beef cattle (35.9%) than among dairy cattle (21.3%), being C. jejuni the species most frequently isolated. PMID:24031386
Fajardo-Cavazos, Patricia; Nicholson, Wayne
As part of an ongoing effort to catalog spore-forming bacterial populations in environments conducive to interplanetary transfer by natural impacts or by human spaceflight activities, spores of Bacillus spp. were isolated and characterized from the interior of near-subsurface granite rock collected from the Santa Catalina Mountains, AZ. Granite was found to contain approximately 500 cultivable Bacillus spores and approximately 10(4) total cultivable bacteria per gram. Many of the Bacillus isolates produced a previously unreported diffusible blue fluorescent compound. Two strains of eight tested exhibited increased spore UV resistance relative to a standard Bacillus subtilis UV biodosimetry strain. Fifty-six isolates were identified by repetitive extragenic palindromic PCR (rep-PCR) and 16S rRNA gene analysis as most closely related to B. megaterium (15 isolates), B. simplex (23 isolates), B. drentensis (6 isolates), B. niacini (7 isolates), and, likely, a new species related to B. barbaricus (5 isolates). Granite isolates were very closely related to a limited number of Bacillus spp. previously found to inhabit (i) globally distributed endolithic sites such as biodeteriorated murals, stone tombs, underground caverns, and rock concretions and (ii) extreme environments such as Antarctic soils, deep sea floor sediments, and spacecraft assembly facilities. Thus, it appears that the occurrence of Bacillus spp. in endolithic or extreme environments is not accidental but that these environments create unique niches excluding most Bacillus spp. but to which a limited number of Bacillus spp. are specifically adapted.
Fajardo-Cavazos, Patricia; Nicholson, Wayne
As part of an ongoing effort to catalog spore-forming bacterial populations in environments conducive to interplanetary transfer by natural impacts or by human spaceflight activities, spores of Bacillus spp. were isolated and characterized from the interior of near-subsurface granite rock collected from the Santa Catalina Mountains, AZ. Granite was found to contain ∼500 cultivable Bacillus spores and ∼104 total cultivable bacteria per gram. Many of the Bacillus isolates produced a previously unreported diffusible blue fluorescent compound. Two strains of eight tested exhibited increased spore UV resistance relative to a standard Bacillus subtilis UV biodosimetry strain. Fifty-six isolates were identified by repetitive extragenic palindromic PCR (rep-PCR) and 16S rRNA gene analysis as most closely related to B. megaterium (15 isolates), B. simplex (23 isolates), B. drentensis (6 isolates), B. niacini (7 isolates), and, likely, a new species related to B. barbaricus (5 isolates). Granite isolates were very closely related to a limited number of Bacillus spp. previously found to inhabit (i) globally distributed endolithic sites such as biodeteriorated murals, stone tombs, underground caverns, and rock concretions and (ii) extreme environments such as Antarctic soils, deep sea floor sediments, and spacecraft assembly facilities. Thus, it appears that the occurrence of Bacillus spp. in endolithic or extreme environments is not accidental but that these environments create unique niches excluding most Bacillus spp. but to which a limited number of Bacillus spp. are specifically adapted. PMID:16597992
Bianchini, Valentina; Borella, Laura; Benedetti, Valentina; Parisi, Antonio; Miccolupo, Angela; Santoro, Eliana; Recordati, Camilla
Thermotolerant Campylobacter spp. are frequently the cause of human gastroenteritis and have assumed more importance in Italy following the increased consumption of raw milk. Our objectives were to determine the prevalence and genotypes of Campylobacter spp. in dairy herds and to investigate the possible sources of bulk milk contamination. Bulk milk from dairy herds (n = 282) was cultured for Campylobacter spp. and Enterobacteriaceae. At three Campylobacter jejuni-positive farms, bovine feces, pigeon intestines, milk, and water points were also investigated. Isolates were identified by PCR and genotyped using multilocus sequence typing (MLST). C. jejuni was detected in 34 (12%) bulk milk samples. The strains belonged to 14 sequence types, and the most common clonal complexes were CC-21, CC-48, and CC-403. No association was demonstrated between the presence of C. jejuni and high levels of Enterobacteriaceae in bulk milk. At the three farms examined, C. jejuni was isolated from bovine feces (25/82 [30.5%]), pigeon intestines (13/60 [21.7%]), bulk milk (10/24 [41.7%]), and water points (4/16 [25%]). MLST revealed lineages that were common between milk and bovine feces but distinct between cattle and pigeons. In one herd, C. jejuni with the same genotype was isolated repeatedly from bulk milk and a cow with an udder infection. Our results showed a high prevalence of C. jejuni in bulk milk and suggested that udder excretion, in addition to fecal matter, may be a route of bulk milk contamination. MLST analysis indicated that pigeons are probably not relevant for the transmission of C. jejuni to cattle and for milk contamination. PMID:24413598
Estepa, Vanesa; Rojo-Bezares, Beatriz; Torres, Carmen; Sáenz, Yolanda
Raw food is a reservoir of Pseudomonas isolates that could be disseminated to consumers. The presence of Pseudomonas spp. was studied in food samples, and the phenotypic and genotypic characterizations of the recovered isolates were analyzed. Two samples of meat (3%, turkey and beef) and 13 of vegetables (22%, 7 green peppers and 6 tomatoes) contained Pseudomonas spp. A total of 20 isolates were identified, and were classified as follows (number of isolates): P. aeruginosa (5), P. putida (5), P. nitroreducens (4), P. fulva (2), P. mosselli (1), P. mendocina (1), P. monteilii (1), and Pseudomonas sp. (1). These 20 Pseudomonas isolates were clonally different by pulsed-field-gel-electrophoresis, and were resistant to the following antibiotics: ticarcillin (85%), aztreonam (30%), cefepime (10%), imipenem (10%), and meropenem (5%), but were susceptible to ceftazidime, piperacillin, piperacillin-tazobactam, doripenem, gentamicin, tobramycin, amikacin, ciprofloxacin, norfloxacin, and colistin. Only one strain (Ps158) presented a class 1 integron lacking the 3' conserved segment. The five P. aeruginosa strains were typed by multilocus sequence typing in five different sequence-types (ST17, ST270, ST800, ST1455, and ST1456), and different mutations were detected in protein OprD that were classified in three groups. One strain (Ps159) showed a new insertion sequence (ISPa47) truncating the oprD gene, and conferring resistance to imipenem.
Carvalho, Fátima C. T.; Sousa, Oscarina V.; Carvalho, Edirsana M. R.; Hofer, Ernesto; Vieira, Regine H. S. F.
This study investigated the presence and antibiotic resistance of Salmonella spp. in a shrimp farming environment in Northeast Region of Brazil. Samples of water and sediments from two farms rearing freshwater-acclimated Litopenaeus vannamei were examined for the presence of Salmonella. Afterwards, Salmonella isolates were serotyped, the antimicrobial resistance was determined by a disk diffusion method, and the plasmid curing was performed for resistant isolates. A total of 30 (16.12%) of the 186 isolates were confirmed to be Salmonella spp., belonging to five serovars: S. serovar Saintpaul, S. serovar Infantis, S. serovar Panama, S. serovar Madelia, and S. serovar Braenderup, along with 2 subspecies: S. enterica serovar houtenae and S. enterica serovar enterica. About twenty-three percent of the isolates were resistant to at least one antibiotic, and twenty percent were resistant to at least two antibiotics. Three strains isolated from water samples (pond and inlet canal) exhibited multiresistance to ampicillin, tetracycline, oxytetracycline, and nitrofurantoin. One of them had a plasmid with genes conferring resistance to nitrofurantoin and ampicillin. The incidence of bacteria pathogenic to humans in a shrimp farming environment, as well as their drug-resistance pattern revealed in this study, emphasizes the need for a more rigorous attention to this area. PMID:24455280
Carvalho, A A; Cardoso, L L; Nogueira, H S; Menezes, E V; Xavier, M A S; Barreto, N A P; Fernandes, L F; Xavier, A R E O
Acinetobacter sp isolates deserve special attention once they have emerged globally in healthcare institutions because they display numerous intrinsic and acquired drug-resistance mechanisms. This study assessed the antibiotic susceptibility profile, the presence of the genetic marker blaOXA-23, and the clonal relationship among 34 nosocomial isolates of Acinetobacter spp obtained at a hospital in southeastern Brazil. Antibiotic sensitivity analysis was performed by the standard disc-diffusion method. All isolates were found to be extensively resistant to several drugs, but sensitive to polymyxin B. A polymerase chain reaction (PCR) assay was used to detect the blaOXA-23 gene, which is associated with carbapenem resistance. The genetic profile and the clonal relationship among isolates were analyzed via enterobacterial repetitive intergenic consensus (ERIC)-PCR. The Acinetobacter spp were divided into four groups with 22 distinct genetic subgroups. ERIC-PCR analysis revealed the genetic diversity among isolates, which, despite having a heterogeneous profile, displayed 100% clonality among 56% (19/34) of them.
Zhang, Ping; Hozalski, Raymond M; Leach, Lynne H; Camper, Anne K; Goslan, Emma H; Parsons, Simon A; Xie, Yuefeng F; LaPara, Timothy M
Haloacetic acids are a class of disinfection byproducts formed during the chlorination and chloramination of drinking water that have been linked to several human health risks. In this study, we isolated numerous strains of haloacetic acid-degrading Afipia spp. from tap water, the wall of a water distribution pipe, and a granular activated carbon filter treating prechlorinated water. These Afipia spp. harbored two phylogenetically distinct groups of alpha-halocarboxylic acid dehalogenase genes that clustered with genes previously detected only by cultivation-independent methods or were novel and did not conclusively cluster with the previously defined phylogenetic subdivisions of these genes. Four of these Afipia spp. simultaneously harbored both the known classes of alpha-halocarboxylic acid dehalogenase genes (dehI and dehII), which is potentially of importance because these bacteria were also capable of biodegrading the greatest number of different haloacetic acids. Our results suggest that Afipia spp. have a beneficial role in suppressing the concentrations of haloacetic acids in tap water, which contrasts the historical (albeit erroneous) association of Afipia sp. (specifically Afipia felis) as the causative agent of cat scratch disease.
Nardoni, S; Merildi, V; Frangioni, S; Ariti, G; Verin, R; Vannucci, P; Mancianti, F
Malassezia spp. genus is represented by several lipophilic yeasts, normally present on the skin of many warm-blooded vertebrates, including man. Swine are one of the less investigated animal species. The aim of the present work was to study the occurrence of Malassezia spp. in the external ear canal of 408 healthy swine of different breeds, under different breeding conditions. For this purpose N. 185 free-ranging wild boars, N. 107 large size pigs and 116 Cinta Senese breed were selected. Animals were of both genders, with age ranging from 8 months to 4 years. The subjects were culturally and molecularly checked for Malassezia spp. Ninety-two out of 408 animals scored positive for Malassezia yeasts (22.5%). Malassezia pachydermatis, Malassezia sympodialis and Malassezia furfur were recognized. M. pachydermatis was the sole species isolated from wild boars (12.9%), Cinta Senese (20.7%) and juvenile large size pigs (13.6%); 88% of large size breeds adult subjects scored positive for M. sympodialis (63.6%) and M. furfur (22.7%), respectively. The study focus on scarcely investigated epidemiological aspects of Malassezia spp. in this animal species.
Wuthiekanun, Vanaporn; Amornchai, Premjit; Paris, Daniel H.; Langla, Sayan; Thaipadunpanit, Janjira; Chierakul, Wirongrong; Smythe, Lee D.; White, Nicholas J.; Day, Nicholas P. J.; Peacock, Sharon J.
Pathogenic Leptospira spp., the causative agents of leptospirosis, are slow-growing Gram-negative spirochetes. Isolation of Leptospira from clinical samples and testing of antimicrobial susceptibility are difficult and time-consuming. Here, we describe the development of a new solid medium that facilitates more-rapid growth of Leptospira spp. and the use of this medium to evaluate the Etest's performance in determining antimicrobial MICs to drugs in common use for leptospirosis. The medium was developed by evaluating the effects of numerous factors on the growth rate of Leptospira interrogans strain NR-20157. These included the type of base agar, the concentration of rabbit serum (RS), and the concentration and duration of CO2 incubation during the initial period of culture. The highest growth rate of NR-20157 was achieved using a Noble agar base supplemented with 10% RS (named LVW agar), with an initial incubation at 30°C in 5% CO2 for 2 days prior to continuous culture in air at 30°C. These conditions were used to develop the Etest for three species, L. interrogans (NR-20161), L. kirschnerii (NR-20327), and L. borgpetersenii (NR-20151). The MICs were read on day 7 for all samples. The Etest was then performed on 109 isolates of pathogenic Leptospira spp. The MIC90 values for penicillin G, doxycycline, cefotaxime, ceftriaxone, and chloramphenicol were 0.64 units/ml and 0.19, 0.047, 0.5, and 2 μg/ml, respectively. The use of LVW agar, which enables rapid growth, isolation of single colonies, and simple antimicrobial susceptibility testing for Leptospira spp., provides an opportunity for new areas of fundamental and applied research. PMID:23114772
Campylobacter spp. has been identified as one of the leading causative agents of food borne diarrheal illness. Epi-demiological evidence has shown that poultry is the main source for human infection. Currently there are no consistently effective treatments to eliminate Campylobacter from poultry flo...
Asato, Valeria C; Vilches, Viviana E; Pineda, María G; Casanueva, Enrique; Cane, Alejandro; Moroni, Mirian P; Brengi, Silvina P; Pichel, Mariana G
Cronobacter species are opportunistic pathogens associated with severe infections in neonates and immunocompromised infants. From January 2009 through September 2010, two cases of neonatal infections associated with Cronobacter malonaticus and one case associated with Cronobacter sakazakii, two of them fatal, were reported in the same hospital. These are the first clinical isolates of Cronobacter spp. in Argentina. The objective of this work was to characterize and subtype clinical isolates of Cronobacter spp. in neonate patients, as well as to establish the genetic relationship between these isolates and the foodborne isolates previously identified in the country. Pulsed-field gel electrophoresis analysis showed a genetic relationship between the C. malonaticus isolates from two patients. Different results were found when the pulsed-field gel electrophoresis patterns of clinical isolates were compared with those deposited in the National Database of Cronobacter spp.
Taboada, Eduardo N.; Acedillo, Rey R.; Carrillo, Catherine D.; Findlay, Wendy A.; Medeiros, Diane T.; Mykytczuk, Oksana L.; Roberts, Michael J.; Valencia, C. Alexander; Farber, Jeffrey M.; Nash, John H. E.
We have used comparative genomic hybridization (CGH) on a full-genome Campylobacter jejuni microarray to examine genome-wide gene conservation patterns among 51 strains isolated from food and clinical sources. These data have been integrated with data from three previous C. jejuni CGH studies to perform a meta-analysis that included 97 strains from the four separate data sets. Although many genes were found to be divergent across multiple strains (n = 350), many genes (n = 249) were uniquely variable in single strains. Thus, the strains in each data set comprise strains with a unique genetic diversity not found in the strains in the other data sets. Despite the large increase in the collective number of variable C. jejuni genes (n = 599) found in the meta-analysis data set, nearly half of these (n = 276) mapped to previously defined variable loci, and it therefore appears that large regions of the C. jejuni genome are genetically stable. A detailed analysis of the microarray data revealed that divergent genes could be differentiated on the basis of the amplitudes of their differential microarray signals. Of 599 variable genes, 122 could be classified as highly divergent on the basis of CGH data. Nearly all highly divergent genes (117 of 122) had divergent neighbors and showed high levels of intraspecies variability. The approach outlined here has enabled us to distinguish global trends of gene conservation in C. jejuni and has enabled us to define this group of genes as a robust set of variable markers that can become the cornerstone of a new generation of genotyping methods that use genome-wide C. jejuni gene variability data. PMID:15472310