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Investigation on thermophilic Campylobacter spp. in commercial poultry farms in Northern Italy
Full text linkCampylobacteriosis, a human infection caused by thermophilic Campylobacter spp. (particularly C. jejuni and C. coli), is the most commonly reported zoonosis in the European Union and one of the main bacterial causes of human gastroenteritis worldwide. Domestic poultry are the main reservoir of these microorganisms and harbour them without clinical manifestations. Handling and consumption of contaminated poultry meat has been reported as main source of human infection. Although campylobacteriosis is generally a self-limiting illness, severe cases require antimicrobial treatment, with macrolides and fluoroquinolones used as first-line therapy, while tetracyclines are second-choice drugs. Nowadays resistance to these antimicrobials is increasingly observed in C. jejuni and C. coli, determining a worldwide concern for public health, since resistant strains can be easily transmitted from animals to humans via the food-chain. Despite the high number of commercial broiler and turkey farms in Northern Italy and the economic importance of this sector in the Italian animal production, few on-farm studies have been carried out to investigate on the colonization of poultry with thermophilic Campylobacter spp. in this area. Considering that, together with the remarkable public health implications of the presence of Campylobacter spp. in poultry farms, the present study was aimed at determining the occurrence, the genetic diversity and the antimicrobial resistance of thermophilic campylobacters in the two leading poultry species reared for meat production in Italy, i.e. broilers and meat turkeys. A total of 10 broiler flocks reared in 7 different farms in Veneto and Lombardy regions and 3 turkey flocks belonging to 3 farms located in Veneto region were examined for the presence of thermophilic Campylobacter spp. by collecting cloacal swabs from randomly selected birds. Broiler flocks were sampled once, while turkeys were monitored with several samplings for the whole production cycle in order to evaluate the development of the colonization over time, because it has not been deeply studied in turkeys. Campylobacter isolates were genetically characterized by flaA-SVR typing and then examined for their phenotypic antimicrobial resistance to a wide range of antimicrobial drugs by the disk diffusion method. The occurrence of class 1 and class 2 integrons was also investigated. A wide colonization by C. jejuni and C. coli was observed both in broiler flocks (7 positive out of 10) and in turkey flocks (all positive), with differences among flocks in the Campylobacter species distribution, although an overall dominance of C. jejuni was detected. Interestingly, the onset of colonization differed considerably among turkey flocks, but in all of them it persisted up to the time of slaughter. A high genetic diversity among isolates was detected both in broilers and turkeys, with most genotypes showing a flock- or farm-specificity. Most strains isolated from both avian species were resistant to a high number of antimicrobials, including those of first and second choice for human campylobacteriosis therapy. Conversely, either class 1 or class 2 integrons were not detected. However, it should be noted that differences between broilers and turkeys were detected both in genotypes colonizing them and in antimicrobial resistance, suggesting discrepancies among the two poultry species in the epidemiological dynamics of Campylobacter spp. Overall, this study provides new insights into the epidemiology of thermophilic Campylobacter spp. in commercial broilers and meat turkeys in Northern Italy and suggests that they may represent a concrete risk for the transmission of multidrug-resistant strains to humans in our countryLa campilobatteriosi, un’infezione umana provocata dalle specie termofile di Campylobacter (soprattutto da C. jejuni a C. coli), è la zoonosi più frequentemente riportata nell’Unione Europea e una delle principali cause di gastroenterite batterica nell’uomo a livello mondiale. Le specie avicole domestiche sono il principale reservoir di questi microrganismi, dei quali sono carrier asintomatici, e infatti la manipolazione e il consumo di carne avicola contaminata rappresentano le principali fonti d’infezione per l’uomo. Nonostante la campilobatteriosi sia una malattia autolimitante, nei casi gravi è necessario un trattamento antimicrobico. Macrolidi e fluorochinoloni costituiscono la terapia d’elezione, seguiti dalle tetracicline come molecole di seconda scelta. Attualmente il fenomeno dell’antimicrobico-resistenza è sempre più comune in C. jejuni e in C. coli, e in tutto il mondo rappresenta motivo di grande preoccupazione per la sanità pubblica, poiché ceppi antimicrobico-resistenti possono essere trasferiti dagli animali all’uomo attraverso gli alimenti. Nonostante l’elevata concentrazione di allevamenti intensivi di polli e tacchini da carne nel Nord Italia e l’importanza economica di questo settore nel comparto delle produzioni animali italiane, sono poche le indagini svolte negli allevamenti di quest’area geografica per valutare la presenza di Campylobacter termofili a livello di produzione primaria. Alla luce di questo e delle importanti implicazioni di sanità pubblica della presenza di questi microrganismi zoonotici negli allevamenti avicoli, questo studio è stato intrapreso allo scopo di valutare la presenza, la diversità genetica e l’antimicrobico-resistenza di Campylobacter termofili nelle due specie avicole da carne maggiormente allevate in Italia: polli broiler e tacchini. In totale sono stati esaminati 10 gruppi di broiler di 7 diversi allevamenti di Veneto e Lombardia, e 3 gruppi di tacchini di 3 allevamenti veneti. In ciascun gruppo di animali sono stati eseguiti tamponi cloacali a soggetti scelti causalmente. In ciascun gruppo di broiler è stato eseguito un campionamento, mentre nei tacchini sono stati svolti numerosi campionamenti per valutare l’andamento della colonizzazione durante tutto il ciclo produttivo, aspetto poco studiato in questa specie avicola. I Campylobacter isolati sono stati caratterizzati geneticamente mediante flaA-SVR typing e quindi sottoposti ad antibiogramma secondo il metodo della diffusione in agar per valutarne la sensibilità a più antimicrobici. Inoltre è stata valutata la presenza di integroni di classe 1 e 2. È stata rilevata una notevole diffusione di C. jejuni e C. coli sia nei broiler (7 gruppi positivi su 10), sia nei tacchini (positività di tutti e 3 i gruppi), con differenze tra i gruppi nella distribuzione delle due specie termofile e una complessiva predominanza di C. jejuni. Nei gruppi di tacchini la positivizzazione è avvenuta in tempi molto diversi, ma in tutti è persistita fino al termine del ciclo produttivo. É stata riscontrata una notevole diversità genetica nei ceppi isolati sia dai broiler, sia dai tacchini e la maggior parte dei genotipi era esclusiva di un gruppo di animali o di un allevamento. Inoltre, la maggioranza dei ceppi isolati da entrambe le specie avicole era resistente a molti antimicrobici, compresi quelli di prima e seconda scelta nella terapia della campilobatteriosi. Al contrario, non sono stati riscontrati integroni né di classe 1 né di classe 2. É importante notare che sono state riscontrate differenze tra broiler e tacchini sia relativamente ai genotipi dai quali erano colonizzati, sia nell’antimicrobico-resistenza, riscontri che suggeriscono il sussistere di differenze tra le popolazioni di Campylobacter che colonizzano l’una e l’altra specie avicola. Nel complesso, questo studio apporta nuovi elementi sull’epidemiologia di Campylobacter spp. in polli e tacchini da carne del Nord Italia, che possono rappresentare un rischio concreto per la trasmissione di ceppi multi-resistenti all’uom
Multilocus Sequence Typing (MLST) of campylobacters to identify potential sources and vehicles of colonization in turkey farms.
No full textA molecular investigation on potential sources and vehicles of Campylobacter infection in commercial turkey farms was carried out. C. jejuni and C. coli strains isolated from breeders, their progeny and the growing flocks reared in 2 different farms for 3 sequential production cycles were analysed by Multilocus Sequence Typing (MLST). Strains isolated from environmental sources (i.e. drinking water, anteroom and in-house floor, flies, Alphitobius diaperinus), collected during the same production cycles, were also analysed. MLST was performed according to the seven-loci schemes employing the primer sets and the experimental conditions suggested by the Campylobacter PubMLST database. Genetic similarity was detected between isolates from breeders and their progeny, as well as between birds and environmental sources, such as flies, Alphitobius diaperinus and drinking water. Results of the present study suggest that these environmental sources may be involved in Campylobacter colonization of meat turkeys
Indagine sulla colonizzazione da Campylobacter termofili in allevamenti di tacchini da carne: risultati preliminari.
No full textSince epidemiology of thermophilic campylobacters in turkeys has been scarcely investigated, we carried out longitudinal studies aimed at identifying potential sources and vehicles of Campylobacter infection in commercial meat turkey farms. A preliminary sampling in turkey breeders flocks was carried out in order to detect a Campylobacter-positive flock. Then, a monitoring of 2 meat turkey farms rearing the progeny of the positive turkey breeders flocks was performed. Meconium was collected from day-old chicks at the hatchery and then faecal swabs were collected three times at the farm (at the beginning, middle and end of the production cycle). Farm sampling consisted of multiple drinking water samples, surface swabs, air, overshoe samples in the anteroom and inside the house, and insects. Sampling was also carried out during downtime. Isolation and identification of Campylobacter spp. were performed by a conventional culture method and a multiplex end-point PCR assay, respectively. Day-old chicks tested negative for thermophilic campylobacters, whereas C. jejuni and C. coli were subsequently isolated from birds throughout the entire cycle in all flocks except for one. Both Campylobacter species were also frequently isolated from drinkers, flies and lesser mealworms. Water, air from inside the house, and surface swabs were always negative. These findings suggest that vertical transmission could not be an important source of flock infection, whereas horizontal transmission should be considered the major route for colonization of turkeys, as campylobacters were found in several environmental sources
Presenza e antibiotico-resistenza di Campylobacter termofili in allevamenti di polli e tacchini
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Pet reptiles as potential reservoir of Campylobacter species with zoonotic potential
No full textContext
Campylobacteriosis is a public health concern worldwide
and several Campylobacter species have been associated
with the disease in people. Companion animal ownership
has been shown to be associated with an elevated
risk of Campylobacter infection, but little is known about
Campylobacter in pet reptiles.
Main conclusion
The present study reports the isolation of Campylobacter
hyointestinalis from reptiles. Pet reptiles could be potential
reservoirs of Campylobacter fetus and C hyointestinalis and thus
a source of infection for people.
Approach
During 2011, a total of 109 cloacal swabs were collected
from captive reptiles (including 49 chelonians, 15 lizards
and 45 snakes) kept in a zoo (42 animals) and in private
households (67 animals) in northern Italy, in order to assess
the presence of Campylobacter species. Campylobacters were
isolated and identified by PCR.
Results
Eight (7.3 per cent) reptiles tested positive for C fetus subspecies
fetus or C hyointestinalis subspecies hyointestinalis.
C fetus subspecies fetus was isolated from four chelonians
(one Chelydra serpentina and three Testudo hermanni hermanni)
and one lizard (Tupinambis rufescens). C hyointestinalis
subspecies hyointestinalis was isolated from one chelonian
(Geochelone sulcata) and two lizards (Iguana iguana). All
snakes tested negative.
Interpretation
The Campylobacter species isolated in the present study are
known to be pathogenic to people. C fetus is reported to
cause gastroenteritis, septicaemia and disseminated infections.
C hyointestinalis has been isolated from both animals
and people and has been shown to be the cause of human
gastroenteritis and septicaemia. C fetus colonisation has previously
been reported in human-raised and pet shop reptiles.
Significance of findings
In contrast to other pet species, there is no perception of
reptiles as potential shedders and spreaders of Campylobacter.
The results of the present study suggest that the risk of
acquiring Campylobacter infection should be considered
when keeping reptiles as companion animals. Further
research is needed to better understand the distribution of
C fetus and C hyointestinalis, their excretion rate in reptiles
and their pathogenic potential for humans
Indagine sulla presenza e la sensibilità agli antibiotici di Campylobacter termofili isolati da polli broiler in Nord Italia
No full textThermophilic Campylobacter spp. infections are the main cause of human bacterial gastroenteritis worldwide, and currently representing a relevant public health problem. Furthermore, an increasing number of thermophilic Campylobacter isolated from animals, humans and foodstuffs is resistant to antimicrobial drugs commonly used in therapy of human campylobacteriosis. Since the leading role of poultry in transmitting the infection to humans, this study was carried out to evaluate the presence and the antimicrobial susceptibility of thermophilic Campylobacter spp. in commercial broiler farms in Northern Italy. A total of 200 cloacal swabs from 10 chicken flocks were examined. Seven chicken flocks were positive for thermophilic Campylobacter. C. jejuni was detected in 63.8% of positive samples and C. coli in 36.2%. The agar disk diffusion method (Kirby-Bauer, 1966) was used to test the susceptibility to 20 antimicrobial drugs of 36 C. jejuni isolates and 21 C. coli isolates. All isolates showed resistance to multiple antimicrobial drugs. Most isolates were resistant to quinolones, ampicillin and cephalosporins. A number of isolates was also resistant to oxytetracycline and sulphametoxazole+trimethoprim. Most isolates were susceptible to aminoglycosides, amoxicillin+clavulanic acid and cefotaxime, and all susceptible to chloramphenicol. Susceptibility to macrolides, clindamycin, oxytetracycline and streptomycin was different between C. coli and C. jejuni. Particularly, most of C. coli isolates were resistant to macrolides and clindamycin, while most C. jejuni were susceptible. This study showed a widespread presence of thermophilic Campylobacter in commercial broilers and a high occurrence of antimicrobial resistance among them. Chicken meat represents one of the main sources of food-borne infection in humans and antimicrobial resistant isolates could be transmitted to humans through food. Therefore, antimicrobial resistance profiles of Campylobacter strains should be strictly monitored due to the relevant impact on public health
Prima segnalazione della presenza di integroni di classe 1 e 2 in E. coli isolati da tacchini da carne
No full textThe aim of this study was to determine the presence of class 1 and class 2 integrons in multi-drug resistant Avian Pathogenic E. coli (APEC) isolated from diseased turkeys. Notoriously, class 1 and class 2 integrons play an important role in the selection of multidrug-resistant bacteria by facilitating the diffusion of antimicrobial resistance genes. Forty-eight multidrug-resistant strains were collected from commercial turkeys affected by colibacillosis and screened for the presence of integrons and associated gene cassettes. Eight isolates harbored class 1 integrons and 9 strains class 2 integrons. The gene cassettes combination most commonly found was dfrA1-aadA1 and dfrA1-sat2-aadA1 in class 1 and class 2 integrons, coding for aminoglycoside and trimethoprim resistance. One class 1 integron positive isolate carried a rare type of gene cassette arrangement (estX-aadA1). This study reports for the first time the presence of class 1 and 2 integrons in Avian Pathogenic E. coli (APEC) isolates from commercial turkeys
A longitudinal study on thermophilic Campylobacter spp. in commercial turkey flocks in Northern Italy: occurrence and genetic diversity
No full textPoultry are recognized as main reservoir of thermophilic campylobacters, but few studies have been carried out on commercial meat turkeys. This study was aimed at assessing the occurrence of thermophilic Campylobacter spp., their genetic diversity and the trend of the infection during the whole production cycle of 3 turkey flocks from different farms in Northern Italy. Flocks were monitored from the housing of day-old poults to slaughter time by collecting samples (meconium and cloacal swabs) at weekly intervals up to the recovery of Campylobacter spp., and then twice a month. A conventional culture method and a multiplex PCR assay were used for Campylobacter detection and identification. A subset of isolates was genetically characterized by RAPD-PCR and flaA-SVR sequencing. Although at different times, all flocks became colonized by C. jejuni and/or C. coli, which persisted throughout the whole production cycle. Overall, 9 RAPD types and 14 flaA-SVR types were detected, with differences in their distribution among flocks and sampling times. Moreover, changes in Campylobacter genotypes colonizing turkeys were observed over time within each flock. These findings suggest that Italian commercial turkeys might be widely colonized by different genotypes of C. jejuni and C. coli and differences in the distribution and epidemiological dynamics of these microorganisms might occur among flocks
Absence of class 1 and class 2 integrons among Campylobacter jejuni and Campylobacter coli isolated from poultry in Italy.
No full textThe paper reports on a study carried out in order to determine the presence of class 1 and 2 integrons in Campylobacter jejuni and Campylobacter coli isolated from intensively reared chickens and turkeys.
Despite the large consumption of chicken and turkey meat worldwide, the recognized role of poultry as reservoir of thermophilic campylobacters and the increasing of antimicrobial resistance in campylobacters, very few studies have been carried out to detect the presence of these genetic structures.
Although evidence of class 1 integron-carrying Campylobacter exists, no integrons were detected in our study. Therefore we can assume that the horizontal gene transfer mediated by integrons may not represent a significant mechanism for the dissemination of antimicrobial resistance determinants in Campylobacter organism
Antimicrobial resistance of Campylobacter jejuni and Campylobacter coli from poultry in Italy.
No full textThis study aimed at assessing the antimicrobial resistance (AMR) of Campylobacter isolates from broilers and turkeys reared in industrial farms in Northern Italy, given the public health concern represented by resistant campylobacters in food-producing animals and the paucity of data about this topic in our Country. Thirty-six C. jejuni and 24 C. coli isolated from broilers and 68 C. jejuni and 32 C. coli from turkeys were tested by disk diffusion for their susceptibility to apramycin, gentamicin, streptomycin, cephalothin, cefotaxime, ceftiofur, cefuroxime, ampicillin, amoxicillin + clavulanic acid, nalidixic acid, flumequine, enrofloxacin, ciprofloxacin, erythromycin, tilmicosin, tylosin, tiamulin, clindamycin, tetracycline, sulfamethoxazole + trimethoprim, chloramphenicol. All broiler strains and 92% turkey strains were multidrug resistant. Very high resistance rates were detected for quinolones, tetracycline and sulfamethoxazole + trimethoprim, ranging from 65% to 100% in broilers and from 74% to 96% in turkeys. Prevalence of resistance was observed also against ampicillin (97% in broilers, 88% in turkeys) and against at least 3 cephalosporins (93-100% in broilers, 100% in turkeys). Conversely, no isolates showed resistance to chloramphenicol and tiamulin. Susceptibility prevailed for amoxicillin + clavulanic acid and aminoglycosides in both species; for macrolides and clindamycin among turkey strains and among C. jejuni from broilers, while most C. coli strains from broilers (87,5%) were resistant. Other differences between C. jejuni and C. coli were observed markedly in broiler isolates, with the overall predominance of resistance in C. coli compared to C. jejuni. This study provides updates and novel data on the AMR of broiler and turkey campylobacters in Italy, revealing the occurrence of high resistance to several antimicrobials, especially key drugs for the treatment of human campylobacteriosis, representing a potential risk for public health
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