Drug use and antimicrobial resistance among Escherichia coli and Enterococcus spp. isolates from chicken and turkey flocks slaughtered in Quebec,Canada

An observational study was conducted of chicken and turkey flocks slaughtered at federal processing plants in the province of Quebec, Canada. The objectives were to estimate prevalence of drug use at hatchery and on farm and to identify antimicrobial resistance (AMR) in cecal Escherichia coli and Enterococcus spp. isolates and factors associated with AMR. Eighty-two chicken flocks and 59 turkey flocks were sampled. At the hatchery, the most used antimicrobial was ceftiofur in chickens (76% of flocks) and spectinomycin in turkeys (42% of flocks). Virginiamycin was the antimicrobial most frequently added to the feed in both chicken and turkey flocks. At least 1 E. coli isolate resistant to third-generation cephalosporins was present in all chicken flocks and in a third of turkey flocks. Resistance to tetracycline, streptomycin, and sulfisoxazole was detected in > 90% of flocks for E. coli isolates. Antimicrobial resistance (AMR) was observed to bacitracin, erythromycin, lincomycin, quinupristin-dalfopristin, and tetracycline in both chicken and turkey flocks for Enterococcus spp. isolates. No resistance to vancomycin was observed. The use of ceftiofur at hatchery was significantly associated with the proportion of ceftiofur-resistant E. coli isolates in chicken flocks. In turkey flocks, ceftiofur resistance was more frequent when turkeys were placed on litter previously used by chickens. Associations between drug use and resistance were observed with tetracycline (turkey) in E. coli isolates and with bacitracin (chicken and turkey), gentamicin (turkey), and tylosin (chicken) in Enterococcus spp. isolates. Further studies are needed to provide producers and veterinarians with alternative management practices and tools in order to reduce the use of antimicrobial feed additives in poultry.     

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.     

Invoices and farmer interviews for vaccine-exposure measurement in turkey-broiler production

Exposure measurement in pharmaco-epidemiological studies can be based on various sources that do not always concur. However, reliability is an important criterion when selecting the method used to assess exposure and interpreting the results obtained. An analysis based on invoices might be more informative and more accurate to assess vaccines exposure (yes/no) in turkey broiler production than a questionnaire administered to farmers, which is nevertheless more feasible and less time-consuming. We compared the two methods to assess vaccination exposure in 239 turkey broiler flocks reared in 129 farms in 2000–2001. The agreement (crude agreement and kappa) was calculated, and association between discrepancy and farm and flock characteristics was investigated.

Marek's-disease vaccine, Newcastle-disease vaccine, turkey haemorrhagic-enteritis vaccine and turkey-rhinotracheitis vaccine exposures were reported on the questionnaire for 2.1, 27.6, 93.0, and 98.3% of flocks, respectively, and for 2.1, 29.3, 89.4, and 86.6%, of invoices for the flocks studied, respectively. A discrepancy was observed in 24.9% of flocks. A discrepancy was observed more frequently in specialised farms without any other animal production (OR = 3.6; CI = 1.5, 8.9) and when the farmer did not know whether vaccination had been performed in the hatchery (OR = 7.1; CI = 2.6, 19.7).     

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.     

Impact of antimicrobial drug usage measures on the identification of heavy users, patterns of usage of the different antimicrobial classes and time-trends evolution

Several measurement units are available to quantify antimicrobial usage in veterinary medicine, to obtain diverse measures such as the weight of active substance used, the live weight (LW) treated, the fraction of animals exposed, the number of treatments recorded or the cost represented. These measures can be applied to study practices variability between farms, to characterize patterns of usage of the different antimicrobial classes or to follow evolution of antimicrobial usage with time. An investigation was carried out to specifically explore the influence of measurement units on the conclusions obtained from such studies. Antimicrobial exposure was explored in a sample of turkey and chicken broiler flocks, using six different units [kg of active compound, treatments, days of administration, kg of LW treated, animal daily dose to treat 1 kg of LW (ADD_kg) and euros] to compare flocks usage variability and patterns of use of the different antimicrobial classes. Time-trends evolutions of macrolides usage in turkey broilers, characterized by percentage of flocks exposed and LW treated, were also compared. In all analyses, the measure gave different results without equivalences, highlighting the necessity for care in choosing the measurement unit and caution in interpreting the figures obtained.     

An original system for the continuous monitoring of antimicrobial use in poultry production in France

Data collection at the flock level on a continuous basis, for antimicrobial use surveillance purposes, provides a complex challenge. Using existing on-farm records, produced as part of the regulations for meat inspection, a sampling scheme was implemented to collect on a simple, representative and continuous basis, the forms providing history of antimicrobial consumption of poultry flocks slaughtered in Brittany. Relational database-stored information provided by the forms includes treatment purpose, antimicrobial drug administered, age at treatment and treatment duration. The aim of this study was to collect information describing antimicrobial drug use quantitatively and qualitatively, by computing the frequency of antimicrobial use according to species, therapeutic class, treatment purpose, duration, and animal age at treatment. A study was carried out to check the validity of farmers' declarations. Examples of data and results are presented such as the relative importance of the different antimicrobial classes in relation to the live weight treated, which shows that beta-lactams and tetracyclines are the main classes used in broilers and turkeys respectively. Average bird weight at treatment was 0.5 kg for a broiler and 3 kg for a turkey. Nevertheless, several 'at-risk' periods could be identified. Initial results after 1 year of data collection showed that this surveillance system provides valid, detailed and useful data about antimicrobial drug use in poultry production at a low cost and with only a moderate time involvement.     

Risk Factors Associated with Detection of Salmonella in Broiler Litter at the Time of New Flock Placement

In this study, we investigated risk factors associated with the probability to detect Salmonella in samples of litter collected within 2 h prior to new flock placement in 76 grow‐out houses on 38 conventional broiler farms located in the US states of Mississippi, Alabama and Texas. We evaluated characteristics of location and layout of the farm; area adjacent to and surrounding the house; house construction; condition and type of equipment in the house; litter management and other production, sanitation, visitation and biosecurity practices; non‐broiler animal species on the farm; and weather conditions on the 3 days leading up to flock placement. Logistic regression was used to model the relationships between probability to detect Salmonella in litter and potential risk factors. In the screening process, each risk factor was evaluated as a single fixed effects factor in a multilevel model that accounted for variability among the sampled farms and their production complexes and companies. Of almost 370 risk factors screened, 24 were associated with the probability to detect Salmonella in litter. These were characteristics of the surroundings of the house, house construction and conditions, litter management, length of downtimes between flocks in the house, biosecurity and farm location. After investigation of collinearity between these variables and building of models for important risk factor categories, the list of candidate variables for the final model was refined to eight factors. The final model demonstrated that a higher probability of detecting Salmonella in litter was strongly associated with the use of wood to construct the base of the walls or to cover the inside of the broiler house foundation, and with the use of fresh wood shavings to top‐dress or completely replace the litter between flocks.     

Risk Factors Associated with Salmonella Status of Broiler Flocks Delivered to Grow‐Out Farms

In a prospective field observational study in the southeastern USA, we sampled gastrointestinal (GI) tracts from chicks of 65 broiler flocks delivered to conventional grow‐out farms for rearing. The flocks were hatched at seven broiler hatcheries. The mean within‐flock prevalence of Salmonella‐positive samples was 6.5% and ranged from 0% to 86.7%. Of the 65 flocks studied, 25 (38.5%) had at least one Salmonella‐positive sample. Accounting for confounding variability among the hatcheries and broiler companies, we tested whether the probability of detecting Salmonella in GI tracts of the chicks delivered was associated with certain characteristics of parent breeder flocks; hatchery production volume; hatchery ventilation system; hatchery egg‐room conditions; egg incubation, candling, hatching, eggshell and bird separation, and bird‐processing procedures; management of hatchery‐to‐farm transportation; day of week of hatch; weather conditions during transportation; or season of the hatch. Two risk factor models were adopted. The first model indicated that a greater number of parent flocks, manual separation of eggshell and bird, and a greater amount of fluff and feces on tray liners used during hatchery‐to‐farm transportation at delivery were associated with increased probability of detecting Salmonella in chick GI tracts, whereas a greater number of birds in the delivery vehicle was associated with decreased probability. The second model indicated that broiler flocks hatched on Tuesdays versus either Mondays or Thursdays (with no hatches on Wednesdays, Fridays or week‐ends), increased average hatchability of the eggs from the parent flocks, and greater amounts of fluff and feces on the transport tray liners at delivery were all associated with increased probability of detecting Salmonella in chick GI tracts. The results of this study suggest potential management decisions to lessen Salmonella contamination of broilers supplied by commercial hatcheries and areas for further research.     

Reduced incidence of Clostridium perfringens-associated lesions and improved performance in broiler chickens treated with normal intestinal bacteria from adult fowl

The dosing of young chicks with cultures of normal gut flora has been termed "competitive exclusion" (CE). This study was undertaken to examine, under field conditions, the effect of CE treatment on counts of intestinal Clostridium perfringens (CP) and on the occurrence of CP-associated disease in broiler chickens. A farm having recurrent CP-associated health problems was selected as study site. The study comprised four broiler houses, with one treated and one untreated flock per house. Treated birds were sprayed with the CE product Broilact upon arrival at the farm. All flocks were offered feed containing the ionophorous anticoccidial agent narasin. The feed did not contain growth promoters. Treatment was associated with positive but statistically nonsignificant effects on gut health. Delayed intestinal proliferation of CP and delayed appearance of CP-associated gut lesions were found in CE-treated flocks. This delay was associated with improved production performance at slaughter.     

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.     

A retrospective study on salmonella infection in Danish broiler flocks

A retrospective longitudinal study was conducted to identify risk factors associated with Salmonella enterica infection in Danish broiler production. The study was based on information in the antemortem database (AM database) where data were available for all broiler flocks slaughtered over the 2-year period from 1992 to 1993 in Denmark. The AM database contains information collected by the ante-mortem veterinarians, from the slaughterhouses, and from the salmonella examinations carried out at the National Veterinary Laboratory. The epidemiological unit was the individual broiler flock. The salmonella status of the flock was determined by examining the caecal tonsils from 16 3-week-old chickens from each flock. This procedure would detect a salmonella-infected flock, with a probability above 95%, if the prevalence is above 20%. Furthermore, the structure and quality of the collected data have been evaluated.

Fourteen variables were selected for analysis by multivariable logistic regression. An increased risk of salmonella infection in the broiler flocks was associated with the biggest hatcheries and feedmill, with an increasing number of houses on the farm, if the preceding flock was infected, and if the flock was reared in the autumn. Additionally, the main variables of the model were analysed by including a random effect at the house level. This resulted only in minor changes of the parameter estimates.     

A field study of naturally occurring specific antibodies against Clostridium perfringens alpha toxin in Norwegian broiler flocks

Necrotic enteritis (NE), a disease associated with high numbers of the intestinal bacterium Clostridium perfringens, is common in intensive broiler production. Antimicrobial feed additives may control the disease, but their use is now being questioned in many countries. A field study was undertaken at the end of 1997 to study the level of naturally occurring specific humoral immunity against phospholipase C (PLC; C perfringens alpha toxin) in Norwegian broiler flocks. Blood samples were collected at hatch from 61 study flocks, and the sampling was repeated for 56 of the same flocks at processing. The level of specific antibodies against PLC was analyzed in an enzyme-linked immunosorbent assay (ELISA) test. Data on production performance and weekly mortality were recorded. The relationship between the age of the hens and the level of specific maternal antibodies in the progenies was studied. The association between the level of the maternal antibodies and the production performance, including mortality, was analyzed. The level of specific antibodies against PLC in day-old broiler flocks was relatively high and varied considerably compared with the levels in the broilers at processing. The progenies from the oldest hens had significantly higher levels of specific antibodies than the chicks from younger hens. No outbreak of NE occurred during the study period, making it impossible to analyze the association between naturally occurring specific immunity against PLC and the occurrence of the disease. However, the results showed that the flocks with high titers of specific maternal antibodies against PLC had lower mortality during the production period than flocks with low titers.     

A longitudinal study of Campylobacter distribution in a turkey production chain

Background

Campylobacter is the most common cause of bacterial enteritis worldwide. Handling and eating of contaminated poultry meat has considered as one of the risk factors for human campylobacteriosis.Campylobacter contamination can occur at all stages of a poultry production cycle. The objective of this study was to determine the occurrence of Campylobacter during a complete turkey production cycle which lasts for 1,5 years of time. For detection of Campylobacter, a conventional culture method was compared with a PCR method. Campylobacter isolates from different types of samples have been identified to the species level by a multiplex PCR assay.

Methods

Samples (N = 456) were regularly collected from one turkey parent flock, the hatchery, six different commercial turkey farms and from 11 different stages at the slaughterhouse. For the detection of Campylobacter, a conventional culture and a PCR method were used. Campylobacter isolates (n = 143) were identified to species level by a multiplex PCR assay.

Results

No Campylobacter were detected in either the samples from the turkey parent flock or from hatchery samples using the culture method. PCR detected Campylobacter DNA in five faecal samples and one fluff and eggshell sample. Six flocks out of 12 commercial turkey flocks where found negative at the farm level but only two were negative at the slaughterhouse.

Conclusion

During the brooding period Campylobacter might have contact with the birds without spreading of the contamination within the flock. Contamination of working surfaces and equipment during slaughter of a Campylobacter positive turkey flock can persist and lead to possible contamination of negative flocks even after the end of the day''s cleaning and desinfection. Reduction of contamination at farm by a high level of biosecurity control and hygiene may be one of the most efficient ways to reduce the amount of contaminated poultry meat in Finland. Due to the low numbers of Campylobacter in the Finnish turkey production chain, enrichment PCR seems to be the optimal detection method here.     

Campylobacter spp. in broiler flocks at farm level and the potential for cross-contamination during slaughter

Screening of broiler flocks for their Campylobacter carriage on farm level and consequently the spread of Campylobacter spp. during slaughtering can help to identify hygiene control points. Therefore, between December 2001 and August 2002 in total 51 broiler flocks from three farms of different geographical regions in Germany were analysed for thermophilic Campylobacter. Campylobacter spp. were isolated from 45% of the broiler flocks examined. Subsequently, 1101 samples were taken from 22 flocks during different stages of processing. Samples were collected from: transport crates before and after cleaning/disinfection, evisceration, post-scalded and post-chilled carcasses and endproducts. Additionally, 45 selected Campylobacter isolates of droppings were genotyped by pulsed-field gel electrophoresis (PFGE). Campylobacter carriage of flocks showed seasonal variation, with the highest contamination rate during the period of June to August. No evidence was found for a horizontal transmission from one broiler flock to the next via a persistent house-contamination. In each positive flock, one to three different genotypes were found. One or two clones dominated isolations obtained from the farm level. The fact that in different flocks indistinguishable isolates of clonal origin were detected during the same rearing period suggested a transmission between the broiler flocks or an intermittent common external source. In one case, isolates of clonal origin were detected in various farms during different rearing periods. Sampling during processing confirmed that the entrance of a positive flock resulted in contamination of the abattoir environment. Campylobacter spp. were isolated from all sampling stages along the processing line, with a percentage of 91.1-100 of isolates at different stages of slaughtering.     

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]).     

Multiple typing for the epidemiological study of contamination of broilers with thermotolerant Campylobacter

This study aims to investigate the genetic diversity of thermotolerant Campylobacter in commercial broiler flocks and in the environment of broiler farms in Belgium. Seven out of 18 investigated flocks became colonized during rearing. Fluorescent amplified fragment length polymorphism (FAFLP), pulsed field gel electrophoresis (PFGE), restriction fragment length polymorphism of the flagellin A gene (flaA-RFLP) and antimicrobial resistance profile (ARP) were used for typing of the isolates. By the combination of FAFLP and PFGE, 22 Campylobacter genotypes could be distinguished. Colonization was almost exclusively with Campylobacter jejuni and unique genotypes were found in each flock. Multiple genotypes were detected in the broilers of 3 flocks, either simultaneously or successively. In 5 flocks, strains that were resistant to at least one antibiotic (mostly tetracycline) were found. The presence of other broiler houses on the farm did not result in a higher probability of colonization. The nipple water was contaminated with the same genotype as the broilers, illustrating its importance for transmission of Campylobacter. The same genotype was detected in a water puddle and in the broiler flock during rearing in 3 flocks. Once, the same genotype was isolated from the ditch water shortly before it was detected in the broilers.     

The dynamics of antimicrobial-resistant Campylobacter jejuni on Japanese broiler farms

We investigated for dynamics of Campylobacter clones on 2 different managerial broiler farms. Campylobacter isolates were differentiated by resistance typing and molecular typing methods. On farm I, the same C. jejuni clones resistant to fluoroquinolone and oxytetracycline were isolated after one and half years again and another susceptible clone was invaded. The susceptible clone was isolated again after half year. Broiler flocks on the farm may be repeatedly infected with a few C. jejuni clones. On farm II, new clones including antimicrobial resistant one, were often invaded. The change of predominant C. jejuni clone in each flock on both the farms was observed, in the absence of antimicrobial selective pressure.     

Quantifying risk factors of coccidiosis in broilers using on-farm data based on a veterinary practice

A study was done to find and quantify risk factors for coccidiosis. The study population consisted of 4774 broiler flocks kept on 177 farms. Flocks were considered a case when at least one bird in the flock showed microscopic presence of oocysts in intestinal scrapings in a grow-out cycle. Other flocks were defined as controls. This was done for three types of Eimeria: Eimeria acervulina, Eimeria tenella and Eimeria maxima. Logistic regression was used to assess variables that influence the occurrence of Eimeria species. There were 49 variables, based on animal, flock or farm level. There was an enhanced risk of coccidiosis due to environmental and management factors that increase the risk of introducing contamination or that are related to hygienic measures. These include lack of use of overalls by visitors, a farmyard which is difficult to clean, bad hygienic status, personnel who might also be working on other farms, presence of other animals on the farm, and feeding and drinking systems which are more difficult to clean. Also, the presence of other diseases on the farm and Eimeria species found in the previous flock increased the risk: of coccidiosis.     

Risk factors for the presence of Campylobacter spp. in Dutch broiler flocks

The campylobacter status of 495 broiler flocks sampled in The Netherlands between April 1997 and December 2000 was related to farm- and flock-specific information obtained from questionnaires to identify potential risk factors for campylobacter presence. Approximately 30% of the flocks tested positive for Campylobacter spp. in at least one pooled faecal sample. Multivariable logistic regression showed significant risk increments for: ages 29-35 days (OR = 2.34) and 36-42 days (OR = 3.96) compared to 22-28 days; > or =5 broiler houses on the premises (OR = 3.02); the presence of other farm animals on the farm (OR = 1.88); the presence of animals on farms within 1 km (OR = 9.56); and summer (OR = 3.48) and fall (OR = 2.59) compared to winter. Furthermore, hatcheries differed (ORs from 5.42 to 20.2), although this variable showed strong collinearity with the variables explaining feed mill and integrated poultry operations. We also identified interaction in which an increased risk from children entering the broiler house (OR = 28.0) was diminished by the use of broiler-specific workclothes. Population attributable fractions (PAFs) suggested that animals on farms within 1 km (PAF = 0.76) and hatchery (PAF = 0.67) had the highest impacts on campylobacter presence in Dutch broiler flocks. These factors, however, lack an easy interpretation of the mechanism behind the suggested effect.     

A prospective study of management and litter variables associated with cellulitis in California broiler flocks

Cellulitis has emerged as an economically important disease of broiler chickens. The impact of environmental risk factors on the incidence of cellulitis has not been evaluated in the United States. Escherichia coli (E. coli), the causative agent, is introduced through skin scratches during the grow out. Our previous work suggested that the litter was an important reservoir for cellulitis-associated E. coli. We hypothesized that factors contributing to a positive environment for E. coli growth would increase the opportunity for exposure of a broiler to an infectious dose of E. coli, capable of initiating a cellulitis lesion. This prospective study of 304 flocks on five farms from two integrated broiler companies was conducted to determine the effect of environmental factors on the prevalence of cellulitis in California broiler flocks. Environmental variables included temperature, wind velocity, and relative humidity (RH) at the litter surface. Litter variables measured included E. coli and total gram-negative bacteria load (colony forming units/g dry matter), water activity, and pH. Management variables such as clean out, the number of flocks reared on the same litter (litter run, LR), and downtime (DT) between flocks were also evaluated. Cellulitis ranged from 0.197% to 6.04%. Significant associations were identified using linear regression between farm, LR, DT, ambient temperature during the brooding period, gram-negative bacteria load in the litter during the brooding period, RH mid-grow out, and E. coli load late in the grow out. The significant variation in the rate of cellulitis between farms combined with the strong association of LR and DT with cellulitis demonstrated that management choices were highly influential in this disease syndrome. Based on these data and our previous findings, managers would be advised to increase DT between flocks and perform a total clean out of the house when a flock processes with a high incidence of cellulitis.