The role of litter beetles as potential reservoir for Salmonella enterica and thermophilic Campylobacter spp. between broiler flocks

We evaluated the role of beetles infesting broiler chicken rearing facilities as potential reservoirs for Salmonella enterica infections between successive broiler flocks. In addition, their role as potential reservoirs for thermophilic Campylobacter spp. was also investigated. Fourteen broiler houses located at 11 different farms were included in the study. The houses were nonrandomly selected on the basis of their salmonella status; nine were persistently contaminated with salmonella whereas five were salmonella negative. For each broiler house, two consecutive broiler flocks (i.e., 28 broiler flocks in all) as well as beetles collected during both rotations of production and in the empty period (after cleaning and disinfection) between these flocks were monitored for the presence of salmonella. Examinations for the presence of campylobacter in the same sample materials were also performed. Beetles sampled during production were positive for salmonella or campylobacter or both. Furthermore, in one house, the occurrence of Salmonella indiana in two consecutive broiler flocks coincided with the presence of S. indiana-contaminated beetles in the empty period between the flocks. The genotype of the identified S. indiana was in all cases identical when analyzed by pulsed-field gel electrophoresis. However, our results also suggest that salmonella from beetles may not always be transmitted to the chickens and that beetles living in contaminated houses can remain free of infection. All cases of campylobacter-positive beetle samples were detected in connection with a positive chicken flock; in no case was campylobacter isolated from beetles taken from the empty period between rotations. Four beetle species were identified during this study. Alphitobius diaperinus was found in all houses and was relatively abundant in most. Typhaea stercorea and Ahasverus advena were found in eight and nine houses, respectively, and were abundant in most of these. Carcinops pumilio was found in small numbers in eight houses. No other insect species was identified. These investigations have shown that beetles in broiler houses infrequently are positive for salmonella. However, transmission of S. indiana between two consecutive broiler flocks can coincide with the presence of salmonella-contaminated beetles in the empty period, indicating that the beetles were the reservoir of S. indiana between the two flocks. Concerning campylobacter, the results suggest that beetles do not play a significant role as a reservoir of campylobacter from one rotation to the next.     

The carry-over of Campylobacter isolates between sequential poultry flocks

The carry-over of Campylobacter strains from one flock to a subsequent flock in the same broiler house has been studied using molecular epidemiological techniques. In all, 524 Campylobacter strains, isolated from two sequential broiler flocks from 60 broiler houses, were typed by restriction fragment polymorphism of the polymerase chain reaction (PCR) product of the flaA and flaB genes (fla typing). Selected strains were also typed using pulsed field gel electrophoresis (PFGE). By fla typing, 15 (21%) of the 60 houses with Campylobacter-positive sequential flocks had identical genotypes. In 10 (16% overall) of these houses the strains were also identical by PFGE. The difference in PFGE patterns in the strains from the three remaining houses may be indicative of genetic instability. Overall, these results suggest that carry-over from one flock to a subsequent flock in the same house is a relatively infrequent event and, therefore, that routine broiler house cleansing and/or disinfection is largely adequate to eliminate Campylobacter contamination. An alternative explanation of the low level carry-over is a persistent source or reservoir, external to the environment of the broiler houses.     

Persistence of salmonella enteritidis in poultry units and poultry food

1. Studies on the survival of Salmonella enteritidis in poultry units and food were carried out over a two‐year period.

2. The organism persisted for at least one year in an empty trial house at the laboratory in which naturally‐infected broiler breeder birds had previously been housed. A similar survival period was seen in a building which had housed an infected layer breeder flock, although infection was not detected in a subsequent pullet flock.

3. Salmonella enteritidis was also frequently found surviving outside poultry houses in small pockets of litter and fan dust which had been left after cleansing and disinfection of the site. On some poultry units S. enteritidis was also found in wild bird droppings.

4. Salmonella contamination appeared to persist preferentially in association with dust particles swept from the floor and in food troughs and S. enteritidis survived at least 26 months in artificially contaminated poultry food.     

Introduction and spread of thermophilic campylobacters in broiler flocks.

Campylobacteriosis is the most commonly reported infectious cause of human gastroenteritis in developed countries and broiler chickens are considered to be the major food-borne source of the infection. The control of the infection in man depends upon its control in broiler flocks but the epidemiology in poultry is poorly understood. Up to 50 per cent of broiler flocks may be infected and most of the birds in an infected flock carry the organisms until slaughter. Vertical transmission through the egg appears unlikely but there are many other potential sources of the infection for the chicks; direct contact with infected animals or birds has been proposed and there is also evidence for indirect transmission through drinking water or insect vectors. It is suggested that the cross-sectional studies discussed in this review should be followed by well designed case-control studies to test the aetiological hypotheses put forward.     

Risk factors for Campylobacter spp. infection in Senegalese broiler-chicken flocks

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.     

Prevalence of antibody to chicken anaemia virus (CAV) in Swedish chicken breeding flocks correlated to outbreaks of blue wing disease (BWD) in their progeny

A serological survey for antibody to Chicken Anaemia Virus (CAV) was performed on broiler breeders as well as layer breeding birds in Sweden at the end of their rearing period. Grandparents (GP) of both types leaving quarantine were in 21 out of 26 cases free from antibody to CAV, but often became infected soon thereafter. A total of 10 outbreaks of blue wing disease (BWD) in 3 series were recorded in the broiler and layer parent generation, all of which were progeny of 3 late seroconverting GP-flocks. All but one of 22 layer parent flocks had been infected and had seroconverted during the rearing period. Subsequently BWD was not recorded from commercial layers. Broiler parent flocks were more protected from CAV infection during rearing. Eighteen out of 94 broiler parent flocks had not developed antibody to CAV before coming into lay. Outbreaks of BWD were reported in progeny flocks from all these broiler breeders, with the exception of those that had been vaccinated. Good hygienic routines along with isolation of the birds delayed the seroconversion to CAV in broiler breeders and vaccination of these breeders protected their progeny from outbreaks of BWD. Broiler flocks in houses where BWD had occurred recently had always antibodies to CAV at slaughter. It was possible to eradicate the infection from the house and prevent the infection between flocks by proper cleaning and disinfection of the broiler houses.     

Risk factors for Listeria monocytogenes contamination in French laying hens and broiler flocks

The objective of this study was to identify potential risk factors for Listeria monocytogenes contamination in French poultry production. Eighty-four flocks of layer hens kept in cages and 142 broiler flocks were included in this study. For each production type, a questionnaire was submitted to farmers and fecal samples were taken to assess the L. monocytogenes status of the flocks during a single visit to the farm. Two logistic regression models (specific to each production) were used to assess the association between management practices and the risk of L. monocytogenes contamination of the flock. The prevalence of L. monocytogenes-positive flocks was 30.9% (95% CI: 21.0; 40.9) and 31.7% (95% CI: 24.0; 39.4) for cage-layers and broiler flocks, respectively. For layer flocks, the risk of L. monocytogenes contamination was increased when pets were present on the production site. When droppings were evacuated by conveyor belt with deep pit storage, the risk of L. monocytogenes contamination decreased significantly. Feed meal was found to be associated with a higher risk of L. monocytogenes contamination than feed crumb. For broiler flocks, the risk of L. monocytogenes contamination was increased when farmers did not respect the principle of two areas (clean and dirty) at the poultry house entrance. A first disinfection by thermal fogging and the absence of pest control of the poultry house before the arrival of the next flock was found to increase the risk of contamination. When litter was not protected during storage and when farm staff also took care of other broiler chicken houses, the risk of L. monocytogenes contamination increased significantly. In the case of the watering system, nipples with cups were found to decrease the risk of contamination.     

Risk factors for Campylobacter spp. contamination in French broiler-chicken flocks at the end of the rearing period.

Our objectives were to identify risk factors for contamination of French broiler flocks by Campylobacter. We used 75 broiler farms in western France. A questionnaire was administered to the farmers and samples of fresh droppings were taken to assess the Campylobacter status of the broiler flocks. 42.7% of the flocks were positive for Campylobacter spp. The risk of contamination of the broiler flocks by Campylobacter was increased in summer/autumn, in houses with static air distribution, when two or more people took care of the flock, in poultry farms with three or more houses and when the drinking water for the chickens was acidified. The presence of litter-beetles in the change room also increased the risk of contamination. The administration of an antibiotic treatment following a disease decreased the risk of a flock being contaminated by Campylobacter.     

A trial of biosecurity as a means to control Campylobacter infection of broiler chickens

We ran a controlled intervention trial to assess whether the risk of a broiler flock becoming infected with Campylobacter could be reduced by biosecurity measures. These were a standard method of cleansing and disinfecting the poultry house prior to stocking, and a standard hygiene protocol followed by all personnel who entered the study house during the flock's life. Thirty-nine flocks were allocated to intervention or control groups in a ratio of 1:2. Intervention flocks were asked to follow the specified biosecurity measures; all flocks were monitored weekly for Campylobacter infection. Analysis of infection at 42 days of age and over the life of the flock showed that the risk of thermophilic Campylobacter infection of broilers was reduced by over 50% in intervention flocks. Parts of the intervention identified as significant in the univariable analysis included twice weekly replenishment of boot dip disinfectant; potential independent risk factors identified included the location of ventilation fans and daily sanitisation of the water supply. The non-random allocation of 10 flocks to the control group may have introduced some study bias (the effect of which is discussed in the paper).     

Economic effects of clinical chicken anemia agent infection on profitable broiler production.

An outbreak of anemia dermatitis syndrome caused by chicken anemia agent (CAA) occurred in 15 broiler flocks. An average of 29% of chickens in these flocks were derived from a common breeder flock. The breeder flock had no antibody to CAA at 20 weeks of age but had seroconverted by 31 weeks. Diseased broiler flocks were derived from eggs laid by the breeder flock between 25 and 30 weeks of age. CAA infection in the breeder flock was subclinical, with no apparent effects on mortality or performance. A strategic program of therapeutic and/or prophylactic antibiotic therapy was begun in affected broiler flocks as soon as the disease was diagnosed. Nevertheless, when the cost of therapy was taken into account, affected broiler flocks had a net income 17.3% to 19.6% lower than normal flocks. Average bird weights were 3.3% to 3.5% lower in affected flocks than in unaffected flocks, and affected flocks had a significantly greater proportion of lighter birds. Average mortality in affected flocks was 2.0% to 2.3% higher than in normal flocks, with peak mortality occurring in the third week of life. There was no apparent effect on feed-conversion ratio.     

Identification of risk factors for the prevalence and persistence of Salmonella in Belgian broiler chicken flocks

According to the European Food Safety Authority, salmonellosis is still one of the main causes of infectious foodborne gastroenteritis in humans. Broilers are an important source of salmonellosis after eggs and pork. Between 1987 and 1999 the trend of human salmonellosis incidence in Belgium increased constantly. However, from 2000 until 2005 a decrease in human cases was observed, probably following the sanitary measures implemented in the poultry breeder and laying sector. In order to decrease human infections it is essential to tackle the problem at the farm level to minimize cross-contamination from farm to fork. This paper seeks to answer two questions: (i) given the Salmonella status of the farm at a certain occasion (equal to the sampling time of the flock), what are the risk factors that the farm will be Salmonella positive at a following occasion? And (ii) what are the risk factors for a farm to be persistently positive for two consecutive flocks? We used surveillance data on 6824 broiler flocks studied for Salmonella infectivity from 2005 to 2006 in Belgium. The farms were tested regularly (3 weeks before slaughter of each broiler flock) for the presence of Salmonella based on multiple faecal samples per flock on a farm yielding clustered data. Generalized estimating equations, alternating logistic regression models, and random-intercept logistic regression models were employed to analyse these correlated binary data. Our results indicated that there are many factors that influence Salmonella risk in broiler flocks, and that they interact. Accounting for interactions between risk factors leads to an improved determination of those risk factors that increase infection with Salmonella. For the conditional analysis, the risk factors found to increase the risk of Salmonella infection on a farm at a current occasion given the previous Salmonella status included: Salmonella infection of day-old chicks (of the current flock); a previously infected flock even though the farm was equipped with a hygiene place to change clothes prior to entering the broiler house; having temporary workmen when there was a separation between birds of different species; and separating birds of different species in the Walloon region relative to the Flanders region. Sanitary measures such as a cleaning and disinfecting procedure conducted by an external cleaning firm, applying the all-in all-out procedure, and hand washing decreased the risk despite their interaction with other factors. From the joint analysis, the most important factors identified for increased risk for persistent Salmonella on a farm involved the interaction between having temporary workmen when there were poultry or farmers in contact with foreign poultry or persons, and the interaction between having temporary workmen when there were poultry or farmers in contact with external poultry or persons.     

Risk factors for Salmonella persistence after cleansing and disinfection in French broiler-chicken houses

A prospective survey was carried out in 86 broiler houses in western France to identify risk factors for Salmonella persistence in French broiler houses. The Salmonella status of the house after cleansing and disinfection, was assessed from gauze-swab samples taken from the walls, feeders, ventilation system and bedding (analysed with classical bacteriological methods). Thirty three (38%) houses had at least one contaminated sample and were classified as Salmonella-contaminated houses in the logistic regression. The absence of a terminal disinfection and a disinfection procedure performed by the farm staff rather than a contractor were positively related to the Salmonella contamination of the house after cleansing and disinfection. The risk for Salmonella persistence after decontamination was increased if rodents were observed by the farmer, if a large part of the access area to the house was accessible to trucks, and if a disease leading to a treatment occurred in the previous flock.     

Risk factors for Salmonella enterica subsp. enterica infection in senegalese broiler-chicken flocks

Our objective was to assess the association of managerial practices, general hygiene and Salmonella infection in Senegalese broiler flocks. Seventy broilers farms were studied from January 2000 to December 2001 around Dakar. A questionnaire was submitted to the farmers and samples of fresh broiler droppings were taken. A 28.6% of the flocks were infected by Salmonella (mainly Hadar and Brancaster serovars). Salmonella infection of the previous flock (OR=6.82) and of day-old chicks (OR=3.73), frequent poultry farmers’ visits (OR=5.38) and keeping sick birds inside the farm (OR=5.32) increased the risk of Salmonella infection. But, using antibiotics on day-old chicks (OR=0.17) and a detergent for cleaning (OR=0.16) decreased the risk.     

Prevalence and risk factors for Campylobacter spp. in chicken broiler flocks in Reunion Island (Indian Ocean)

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]).     

Sources of Salmonellae in broiler chickens in Ontario.

Sources of Salmonellae infecting broiler chicken flocks in Ontario were investigated from July, 1975 to April, 1976. Three broiler flocks were investigated on each of four farms which received chicks from a common hatchery. Samples of feed and new litter were preenriched in nonselective broth subcultured to Salmonella-selective enrichment broth and plated on Salmonella-selective differential agar.Samples of used litter, water, culled chicks, insects, mice, wild birds and environmental swabs were not cultured initially in the nonselective broth. Fecal samples from principal and occasional flock attendants were examined for Samonellae. Salmonella infection, as judged by contaminated flock litter was detected in six flocks on two of the farms while the flocks on the other farms remained negative. Salmonellae were isolated from 23 of 412 feed samples (nine serotypes), six of 35 new wood shaving samples (four serotypes), one of 29 pools of culled chick viscera (one serotype) and 44 of 267 used litter samples (14 serotypes). These results indicate that broiler chicken flocks were infected with diverse Salmonellae introduced in day old chicks, pelleted feeds, wood shavings and residual contamination from the preceding flock.     

A study of the epidemiology of infectious bursal disease in poultry in Queensland, 1976–1979

The epidemiology of infectious bursal disease (IBD) was studied by serology and sometimes by visual examination of the bursa of Fabricius in poultry flocks in Queensland during 1976–1979.
Ten flocks, each of approximately 30,000 meat breeding chickens, were surveyed. All chickens had maternally-derived antibody against IBD virus (IBDV) at hatching and active antibody was not detected while the chickens were brooded on rearing farms. When distributed to breeding farms, 7 of the flocks developed antibody when 11 to 25 weeks of age. The remaining 3 flocks were vaccinated by infection of 10% of the birds and within 4 weeks more than 80% of the chickens had developed precipitating antibody to IBDV.
Blood samples of 20 to 30 broiler chickens were collected at slaughter (7 to 9 weeks of age) from each of 312 broiler flocks raised on 37 contract farms. While the samples from 21 flocks were without detectable antibody to IBDV, all serum samples for 263 flocks contained antibody. The ratio of bursal weight to bodyweight was significantly lower in birds from 144 flocks having antibody to IBDV than in birds from 10 flocks that were without detectable antibody. In sequential studies, IBDV antibody became demonstrable in 27 of 30 flocks when the chickens were one to 6 weeks of age and was accompanied by bursal atrophy.
Serological investigation of 4 flocks of layer breeding chickens on a multi-age farm at approximately monthly intervals resulted in antibody to IBDV being detected at every examination.
Serological tests and bursal examinations were carried out weekly in 2 flocks each of 4000 layer chickens between one and 20 weeks of age. Serum antibody developed in one flock at 4 weeks of age and in the other at 17 weeks of age. In both flocks, bursal atrophy occurred concurrently with the development of antibody.     

Campylobacter spp. in conventional broiler flocks in Northern Ireland: epidemiology and risk factors

Risk factors for Campylobacter infection in conventional broiler flocks in the time period up to the first removal of birds to slaughter were investigated over a maximum of five consecutive production cycles in a cohort of 88 broiler farms in Northern Ireland. Samples for Campylobacter culture, which consisted of 14 cloacal swabs per flock, were collected from one house on each farm prior to the first depopulation of birds. In total 388 flocks were sampled, of which 163 tested positive for Campylobacter spp. (42.0%; 95% CI 35.1–48.9%).

Data on farm and flock variables were obtained from questionnaires and random-effects logistic regression modelling used to investigate the association between these and the Campylobacter status of flocks. Six variables, all of which were significant at p < 0.05, were included in the final multivariable model. These included a combined variable on the presence of rodents on farms, which showed an increased odds of infection in flocks where the farmer reported having observed rodents during the production cycle (OR = 2.1) and/or where rodent droppings were observed at the sampling visit (OR = 2.9). Other variables that were significantly associated with an increased odds of infection included the age of the birds at sampling (odds ratio for its linear effect = 1.16 for each day of increase in age), season (summer versus other seasons OR = 2.0), farms with three or more broiler houses (OR = 2.9 compared to those with one house), the frequency of footbath disinfectant changes (OR = 2.5 for once weekly and OR = 4.0 for less than once weekly compared to twice weekly changes) and a categorical variable on the standard of tidiness and cleanliness of the broiler house ante-room (OR = 2.0 and OR = 4.9 for flocks from houses with poorer standards). There was no significant evidence of direct carry-over of infection from one production cycle to the next, neither was there evidence of other farm species acting as a source of infection.     

Relationship of Campylobacter isolated from poultry and from darkling beetles in New Zealand

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.     

Evaluation of Salmonella serotype distributions from commercial broiler hatcheries and grower houses

By conventional trayliner (hatcheries) and drag swab assembly (broiler houses) culture methods, the isolation distribution of Salmonella serotypes from five commercial broiler hatcheries (three sample times) and 13 broiler farms (eight sample times) was evaluated. A total of 11 different Salmonella serotypes were isolated from hatcheries, with Salmonella heidelberg (9/30) and Salmonella kentucky (6/30) accounting for 50% of the total isolations. Of 700 chick paperpad trayliners sampled, regardless of lot (breeder flock source) or hatchery, 12% were positive for Salmonella. When 10 individual trayliners were cultured from individual lots (same breeder flock source), Salmonella was detected in 24/57 lots (42%). Multiple serotypes were simultaneously isolated from the same lot on three occasions (6%). Of the 21 lots that were serially sampled, the Salmonella serotype detected was different within lots eight times (38%) on at least one occasion of two or more sampling times. Of the 196 individual broiler houses sampled, 44 were positive for Salmonella (42%). Twelve different serotypes were isolated from broiler houses during this study. The serotypes isolated most frequently were S. heidelberg (34/94) and S. kentucky (22/94). These two serotypes accounted for 59.6% (56/94) of the total broiler house isolations. Of the 38 houses that were serially sampled, two or more serotypes were detected in the same broiler house on 20 occasions (53%). Of the 38 serially sampled houses (four or more times), a consistent Salmonella serotype was detected in five houses (13%). In only 5 of the 38 (13%) serially sampled houses did we fail to detect Salmonella on four or more samplings. No significant difference in Salmonella isolation frequency was observed between poultry houses using new or used litter. These data support previous findings indicating that paratyphoid Salmonella serotypes are prevalent in some broiler hatcheries and houses. Further, the observation of multiple serotypes simultaneously and serially isolated from the same breeder hatchery lots suggests that breeder flocks may be infected with more than one serotype, possibly providing a source for multiple serotype infections in progeny grower flocks.