Quantitative Determination of Campylobacter on Broilers along 22 United Kingdom Processing Lines To Identify Potential Process Control Points and Cross-Contamination from Colonized to Uncolonized Flocks.

ABSTRACT: As part of a program to reduce numbers of the human pathogen Campylobacter on retail chickens, 22 broiler processing lines, representing more than 90% of UK production, were characterized by enumerating Campylobacter on pooled neck skins after exsanguination, scalding, defeathering, evisceration, crop removal, inside-outside washing, and air-chilling stages of processing. Sixteen of the processing lines investigated showed significant (P < 0.05) reductions in Campylobacter numbers because of carcass scalding. However, in all of these lines, the following defeathering stage caused a significant increase in Campylobacter contamination that effectively negated the reductions caused by scalding. On four processing lines, primary chilling also caused a significant reduction in numbers of Campylobacter. On three lines, there was a significant microbiological benefit from inside-outside washing. The stages where Campylobacter numbers were reduced require further investigation to determine the specific mechanisms responsible so that the observed pathogen reductions can be optimized and then more widely implemented. The transfer of up to 4 log CFU Campylobacter per g of neck skin from a colonized flock to a following uncolonized flock was observed. Cross-contamination was substantial and still detectable after 5,000 carcasses from an uncolonized flock had been processed. Numbers of Campylobacter recovered from the uncolonized flocks were highest on the first of the uncolonized birds to pass along the line, and in general, the numbers declined as more uncolonized birds were processed. Air sampling recovered low numbers at the processing stages monitored, indicating that airborne transmission was unlikely to be the primary transfer mechanism operating for cross-contamination between flocks.

The association between threatened preterm labour and perinatal outcomes at term: a population-based cohort study.

OBJECTIVE: To estimate the association between threatened preterm labour (TPTL) and perinatal outcomes of infants born at term. DESIGN: A population-based cohort study of perinatal outcomes following TPTL <37 weeks of gestation with delivery at term. SETTING: Nova Scotia, Canada. POPULATION: All non-anomalous, singleton pregnancies ≥37 weeks of gestation without antepartum haemorrhage from 1988 to 2019. METHODS: Using data from the Nova Scotia Atlee Perinatal Database, TPTL was defined as pregnancies with a hospital admission between 20 and 37 weeks of gestation, with a diagnosis code denoting TPTL with administration of antenatal corticosteroids, or with administration of any tocolysis. Poisson regression models were used to estimate the risk ratios (RR) with 95% CI of maternal and perinatal outcomes in women who had an episode of TPTL relative to those who did not. MAIN OUTCOME MEASURES: Birthweight for gestational age below the tenth centile and a composite of perinatal mortality or severe perinatal morbidity. RESULTS: Of 256 599 term deliveries meeting the inclusion criteria, 2278 (0.9%) involved TPTL. The risks of the primary outcomes were higher among those with TPTL relative to those without: birthweight for gestational age below the tenth centile (RR 1.24, 95% CI 1.11-1.39) and the composite of perinatal mortality/severe perinatal morbidity (RR 1.33, 95% CI 1.15-1.54). CONCLUSIONS: Although the prevalence of TPTL in term deliveries is low, affected pregnancies are at increased risk for adverse perinatal outcomes. Increased fetal surveillance should be considered in the management of pregnancies affected by TPTL.

An improved cleaning system to reduce microbial contamination of poultry transport crates in the United Kingdom.

AIM: Following previous research on improving the cleaning of crates used to transport broiler chickens from the farm to the abattoir, a demonstration project was undertaken to investigate improvements in crate washing on a commercial scale. METHODS AND RESULTS: The soak tank of a conventional crate washing system was replaced with a high-performance washer fitted with high-volume, high-pressure nozzles. The wash water could be heated, and a greatly improved filtration system ensured that the nozzles did not lose performance or become blocked. Visual cleanliness scores and microbial counts were determined for naturally contaminated crates which had been randomly assigned to different cleaning protocols. CONCLUSIONS: When a combination of mechanical energy, heat and chemicals (i.e. detergent and disinfectant) was used, the results showed significant improvements to crate cleaning. Reductions of up to 3·6 and 3·8 log10 CFU per crate base were achieved for Campylobacter and Enterobacteriaceae, respectively, along with a marked improvement in visual cleanliness. SIGNIFICANCE AND IMPACT OF THE STUDY: Broiler transport crates may become heavily contaminated with faeces and this may contribute to the spread of disease between farms. The results of this trial may be of use in reducing the spread of zoonotic pathogens in the poultry meat supply chain.

A longitudinal observational study of Salmonella shedding patterns by commercial turkeys during rearing and fattening, showing limitations of some control measures.

1. The onset and progression of Salmonella infections was investigated in commercial turkey flocks from placement at 1 d old until slaughter in "brood and move" systems using a longitudinal observational approach based on faeces and environmental sampling with subsequent culture of Salmonella. 2. Persistent Salmonella Newport contamination was found within rearing houses and on their external concrete aprons after cleaning and disinfection between crops of heavily shedding young birds. 3. Salmonella shedding was often detected by 5 d of age and the frequency of positive samples peaked at 14-35 d. Thereafter Salmonella isolations declined, especially in the later (fattening) stages. Samples were still Salmonella-positive at low prevalence in half of the intensively sampled houses at slaughter age. 4. A number of management interventions to combat Salmonella infection of flocks, including sourcing policy, competitive exclusion cultures and cleaning and disinfection, were inadequate to prevent flock infection, although improved disinfection on one unit was associated with a delay in the onset of flock infection.

Estimating the time at which commercial broiler flocks in Great Britain become infected with Campylobacter: a Bayesian approach.

Campylobacter is a common cause of intestinal disease in humans and is often linked to the consumption of contaminated poultry meat. Despite considerable research on the topic there is a large amount of uncertainty associated with Campylobacter epidemiology. A Bayesian model framework was applied to multiple longitudinal datasets on Campylobacter infection in UK broiler flocks to estimate the time at which each flock was first infected with Campylobacter. The model results suggest that the day of first infection ranges from 10 to 45 days; however, over half had a time of infection between 30 and 35 days. When considering only those flocks which were thinned, 48% had an estimated day of infection within 2 days of the day of thinning, thus suggesting an association between thinning and Campylobacter infection. These results demonstrate how knowledge of the time of infection can be correlated to known events to identify potential risk factors for infection.

Application of a national administrative case definition for the identification of pre-existing diabetes mellitus in pregnancy.

INTRODUCTION: Accurate ascertainment of pregnant women with pre-existing diabetes allows for the comprehensive surveillance of maternal and neonatal outcomes associated with this chronic disease. METHOD: To determine the accuracy of case definitions for pre-existing diabetes mellitus when applied to a pregnant population, a cohort of women who were pregnant in Nova Scotia, Canada, between 1991 and 2003 was obtained from a population-based provincial perinatal database, the Nova Scotia Atlee Perinatal Database (NSAPD). Person-level data from administrative databases using hospital discharge abstract data and outpatient physician services data were linked to this cohort. Various algorithms for defining diabetes mellitus from the administrative data, including the algorithm suggested by the National Diabetes Surveillance System (NDSS), were compared to a reference standard definition from the NSAPD. RESULTS: Validation of the NDSS case definition applied to this pregnant population demonstrated a sensitivity of 87% and a positive predictive value (PPV) of 66.4%. Use of ICD-9 and ICD-10 diagnostic codes among hospitalizations with diabetes mellitus in pregnancy showed important increases in sensitivity and PPV, especially for those pregnancies delivered in tertiary centres. In this population, pregnancy-related administrative data from the hospitalization database alone appear to be a more accurate data source for identifying pre-existing diabetes than applying the NDSS case definition, particularly when pregnant women are delivered in a tertiary hospital. CONCLUSION: Although the NDSS definition of diabetes performs reasonably well compared to a reference standard definition of diabetes, using this definition for evaluating maternal and perinatal outcomes associated with diabetes in pregnancy will result in a certain degree of misclassification and, therefore, biased estimates of outcomes.

Biosecurity-based interventions and strategies to reduce Campylobacter spp. on poultry farms.

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.

Influence of production system on the rate of onset of Campylobacter colonization in chicken flocks reared extensively in the United Kingdom.

1. Because thermophilic Campylobacter spp. are common in chicken flocks reared extensively, cross-sectional and longitudinal studies were carried out on organic and free-range farms to determine the onset of colonisation (lag phase) and likely sources of flock infection. 2. For 14 organic and 14 free range flocks, there was a difference in lag phases, with the former being colonized at a mean of 14·1 d in comparison with 31·6 d for the latter. Whereas most free-range flocks became colonized when released on to pasture, those reared organically were usually colonized at the housed brooding stage. 3. Further study of organic flocks on three farms over 7 successive crop cycles confirmed that colonisation was strongly influenced by the prevailing husbandry conditions and was not a consequence of the length of the rearing period. 4. Molecular epidemiological investigations on a farm showing the shortest lag phase, using PFGE typing with two different restriction enzymes (SmaI and KpnI) and flaA SVR sequence typing, revealed that potential sources of colonisation for organic chickens were already present on the farm at the time of chick placement. Such sources included the ante area of the brooding house, surrounding pasture and other livestock being kept on the farm. 5. Overall, the study demonstrated that, under UK conditions, the prevalence of colonisation was greater in extensive flocks (95-100%) than it was for conventional broilers (55%), similar to the situation in other countries, but all three management systems showed comparable levels of caecal carriage in positive birds (log(10)/g 6·2-6·7).

Effect of delivery method on the efficacy of Salmonella vaccination in chickens.

To investigate whether the efficacy of live vaccines is influenced by the mode of vaccine delivery, a widely-used UK live commercial Salmonella Enteritidis vaccine was delivered to pullet chicks either by spray, in drinking water, or in combination with a bivalent vaccine containing inactivated Salmonella Enteritidis and Salmonella Typhimurium. The birds were subsequently challenged with 10(2) or 10(8) colony-forming units (cfu) of Salmonella Enteritidis through drinking water at either six or 20 weeks of age. Ten days after the challenge, the birds were euthanased and their caecal contents cultured for Salmonella. All of the vaccinated groups contained fewer Salmonella Enteritidis-positive birds than the unvaccinated groups. The 'spray-vaccinated' group contained significantly fewer Salmonella Enteritidis-positive birds than the 'water-vaccinated' group after challenge with 10(8) cfu at 20 weeks. However, there was little or no difference at the other challenge time points between the groups that received vaccine through different modes of delivery.

Salmonella colonisation of laying hens following vaccination with killed and live attenuated commercial Salmonella vaccines.

The aim of this study was to determine the efficacy of a killed Salmonella vaccine and three live vaccines in preventing caecal colonisation of Hy-line Brown pullets by Salmonella Enteritidis PT 4. The lowest number of Salmonella-positive birds following the largest challenge (10(8) cfu) was recorded for live vaccine 1. However, birds treated with the killed vaccine had a significantly lower number of salmonellae in their caeca compared with both the control group and the other vaccine groups (P<0.05).

Low levels of maternal serum PAPP-A in early pregnancy and the risk of adverse outcomes.

OBJECTIVES: To determine if low maternal serum level of pregnancy associated plasma protein A (PAPP-A) measured in early pregnancy can predict adverse pregnancy outcomes and to examine the gestational age (GA) sampling interval for these outcomes. METHODS: This was a nested case-control study from a prospective cohort of women recruited at <20 weeks of gestation in Halifax, NS. Cases (n=248) were defined as women who had a fetal loss or developed preeclampsia, severe pregnancy-induced hypertension (PIH), or small for gestational age infant (SGA). Controls (n=244) were frequency matched to cases by GA at the time of serum sampling (6 to <20 weeks GA). Participant information was obtained from questionnaires and medical chart reviews. RESULTS: Women with a low PAPP-A measure [0.4 MoM). However, performance as a screening test was poor [sensitivity=38.7%; specificity=81.6%; positive likelihood ratio (LR)=2.1; negative LR=0.75]. In the adjusted model, the 10- to 14-week GA period was the only time period where low PAPP-A was significantly associated with adverse outcomes. CONCLUSIONS: Women with a low PAPP-A early in their pregnancy have twice the risk of an adverse outcome, though PAPP-A as a one-time single marker test has limited value.

Effect of ultrasonic treatment during cleaning on the microbiological condition of poultry transport crates.

1. Small sections cut from commercial crates used to transport live poultry to the processing plant were artificially contaminated with effluent taken from a commercial crate-cleaning system. 2. Laboratory trials, involving the immersion of these sections in an ultrasonic water bath (4 kW energy) showed that aerobic plate counts (APC) and counts of Enterobacteriaceae were progressively reduced as the immersion time was increased from 0 to 120 s and the water temperature raised from 35 to 58 degrees C. 3. In subsequent trials at a processing plant, using commercially cleaned crates, there was relatively little effect of ultrasound (or pressure washing) on the biofilm present. However, ultrasonic treatment in combination with an immersion temperature of 60 degrees C reduced counts of Enterobacteriaceae to below the detection limit (log(10) 2.3 cfu) within 1 to 3 min, while APC were reduced by >2 log(10) units after 3 min. 4. It was concluded that ultrasonic treatment has a possible role in the crate-cleaning process, when used in conjunction with higher immersion temperatures. In this way, it could contribute significantly to hygiene control.

Evaluation of the performance of different cleaning treatments in reducing microbial contamination of poultry transport crates.

1. The present systems for cleaning the plastic crates (drawers) used to transport live poultry to the processing plant are known to be inadequate for removing microbial contamination. 2. To investigate possible improvements, a mobile experimental rig was constructed and operated in the lairage of a poultry processing plant. The cleaning rig could simulate the conditions of commercial cleaning systems and utilise freshly emptied crates from the processing plant. 3. The aim of the study was to improve cleaning by enhancing the removal of adherent organic material on the crates and by reducing microbial contamination by at least 4 log(10) units. 4. Trials showed that the most effective treatments against Campylobacter were either (a) the combination of soaking at 55 degrees C, brushing for 90 s, washing for 15 s at 60 degrees C, followed by the application of disinfectant (Virkon S in this study) or (b) the use of ultrasound (4 kW) at 65 degrees C for 3 to 6 min, with or without mechanical brushing of crates. 5. Both of these treatments also achieved a 4 log(10) reduction or more in the counts of Enterobacteriaceae but were less effective in reducing aerobic plate counts. 6. It was noted that there was little correlation between the visual assessment of crate cleanliness and microbiological counts. 7. It was concluded that the demonstrated enhanced cleaning could contribute significantly to overall hygiene control in poultry meat production.

Sources and spread of thermophilic Campylobacter spp. during partial depopulation of broiler chicken flocks.

The practice of partial depopulation or thinning (early removal of a portion of birds from a commercial broiler flock) is a reported risk factor for Campylobacter colonization of residual birds because of the difficulty in maintaining biosecurity during the thinning process. The effect of this practice was studied in detail for 51 target flocks, each at a different growing farm belonging to one of seven major poultry companies throughout the United Kingdom. On 21 of these farms, the target flock was already colonized by Campylobacter, and at slaughter all cecal samples examined were positive, with a mean of 8 log CFU/g. An additional 27 flocks became positive within 2 to 6 days of the start of thinning and had similarly high levels of cecal carriage at slaughter. Just before the thinning process, Campylobacter was isolated frequently from the farm driveways, transport vehicles, equipment, and personnel. Strains from seven farms on which flocks became colonized after thinning were examined by pulsed-field gel electrophoresis typing. An association was found between strains occurring at specific sampling sites and those isolated subsequently from the thinned flocks. The results indicated that particular strains had spread from one farm to another when the farms were jointly owned by the same company and employed the same bird-catching teams and/or vehicles. These results highlight the need for better hygiene control in relation to catching equipment and personnel and more effective cleaning and disinfection of vehicles and bird-transport crates.

Neonatal outcomes with caesarean delivery at term.

OBJECTIVE: To estimate the impact of caesarean delivery on the incidence of selected neonatal outcomes. PATIENTS AND METHODS: A 15-year, population-based, cohort study (1988-2002) using the Nova Scotia Atlee Perinatal Database compared neonatal outcomes in term newborns born by spontaneous and assisted vaginal delivery, with newborns born by caesarean delivery, with and without labour, using multiple logistic regression. RESULTS: From a total of 142 929 deliveries, there were 27 263 caesarean deliveries, 61% of which were performed in labour. Relative risks were adjusted for year of birth, maternal age, parity, smoking, maternal weight at delivery, hypertensive diseases, diabetes, previous caesarean delivery, use of regional anaesthesia, induction of labour, gestational age at delivery and large and small for gestational age, where significant. Caesarean delivery in labour, but not caesarean delivery without labour, had increased risks for depression at birth and neonatal respiratory conditions compared with spontaneous or assisted vaginal delivery. Compared with spontaneous vaginal delivery and assisted vaginal delivery, the risk of major neonatal birth trauma was decreased for infants after caesarean delivery with labour (odds ratio (OR) = 0.34, 95% CI 0.21 to 0.56 and OR = 0.07, 95% CI 0.04 to 0.11, respectively) and caesarean delivery without labour (OR = 0.20, 95% CI 0.08 to 0.52 and OR = 0.04, 95% CI 0.02 to 0.10, respectively). CONCLUSION: Caesarean delivery in labour, compared with vaginal delivery, is more likely to be associated with an increased risk for respiratory conditions and depression at birth than caesarean delivery without labour. Caesarean delivery appears protective against neonatal birth trauma, especially when performed without labour.

Bacteriophage therapy to reduce salmonella colonization of broiler chickens.

Acute enteric infections caused by salmonellas remain a major public health burden worldwide. Poultry, particularly chickens, are known to be the main reservoir for this zoonotic pathogen. Although some progress has been made in reducing Salmonella colonization of broiler chickens by using biosecurity and antimicrobials, it still remains a considerable problem. The use of host-specific bacteriophages as a biocontrol is one possible intervention by which Salmonella colonization could be reduced. A total of 232 Salmonella bacteriophages were isolated from poultry farms, abattoirs, and wastewater in 2004 and 2005. Three phages exhibiting the broadest host ranges against Salmonella enterica serotypes Enteritidis, Hadar, and Typhimurium were characterized further by determining their morphology and lytic activity in vitro. These phages were then administered in antacid suspension to birds experimentally colonized with specific Salmonella host strains. The first phage reduced S. enterica serotype Enteritidis cecal colonization by > or = 4.2 log10 CFU within 24 h compared with controls. Administration of the second phage reduced S. enterica serotype Typhimurium by > or = 2.19 log10 CFU within 24 h. The third bacteriophage was ineffective at reducing S. enterica serotype Hadar colonization. Bacteriophage resistance occurred at a frequency commensurate with the titer of phage being administered, with larger phage titers resulting in a greater proportion of resistant salmonellas. The selection of appropriate bacteriophages and optimization of both the timing and method of phage delivery are key factors in the successful phage-mediated control of salmonellas in broiler chickens.

Campylobacter spp. contamination of chicken carcasses during processing in relation to flock colonisation.

The presence and numbers of campylobacters on chicken carcasses from 26 slaughter groups, originating from 22 single-house flocks and processed in four UK plants, were studied in relation to the level of flock colonisation determined by examining the caecal contents of at least ten birds per group. The prevalence of campylobacters on carcasses from five campylobacter-negative flocks processed just after other negative flocks was low (8.0 log(10) cfu) than carcasses originating from low prevalence flocks (average of 2.3 log(10) cfu; range: <1.1 to 4.1 log(10) cfu). There was a reduction in the numbers of campylobacters on carcasses between plucking and chilling in eight of ten fully colonised flocks. In another eight flocks, a significant (P<0.001) decrease (0.8 log(10) cfu) in the number of campylobacters on carcasses from just before to after chilling was detected. Campylobacter spp. could be isolated from aerosols, particles and droplets in considerable numbers in the hanging-on, defeathering and evisceration areas but not in the chillers. This was the case even when campylobacters were not isolated from the target flock. Campylobacters on carcasses from two partly colonised flocks were either the same subtype, as determined by speciation, Multi-Locus Sequence Typing (MLST) and flaA Restricted Fragment Length Polymorphism (RFLP) typing, as those in the fully colonised flocks processed previously, although not necessarily the most prevalent ones; or were the same subtypes as those found in the caeca of the flock itself. The prevalences of the different campylobacter subtypes found on carcasses from two fully colonised flocks did not closely reflect those found in the caeca. MLST combined with flaA RFLP provided a good method for ascertaining the relatedness of strains isolated from carcasses and caecal contents. This study showed that carcass contamination is related to the within-flock prevalence of campylobacter colonisation, but that contamination from previously processed flocks was also significant, especially on carcasses from low prevalence flocks. Forced dry air cooling of carcasses reduced contamination levels.

A novel method for assessing the role of air in the microbiological contamination of poultry carcasses.

This paper describes a novel method of measuring the contamination of raw foods with airborne bacteria during primary processing. To demonstrate the approach, this study aimed to quantify the role of airborne bacteria in the contamination of broiler chicken carcasses undergoing processing in an evisceration room. Settle plates and broiler carcasses were exposed to the evisceration room air or to ultra-clean air provided by a high efficiency particulate air (HEPA) unit located within the room. The use of ultra-clean air reduced the total aerobic counts on horizontal settle plates by 68-fold, and on vertical settle plates by 14-fold. The use of ultra-clean air had no significant effect on the total aerobic counts on carcasses as measured by sponging (3.5 log(10) CFU cm(-2)) or skin excision (4.0 log(10) CFU cm(-2)). The novel approach was able to show that the carcasses entering the room were so heavily contaminated that the airborne bacteria in the evisceration room contributed less than 1% of the total numbers of bacteria on the carcasses.

Predictors of respiratory depression at birth in the term infant.

OBJECTIVE: To evaluate predictive factors for respiratory depression at birth in infants >/=37 weeks. DESIGN: A population-based cohort study of respiratory depression at birth at term and post-term. SETTING: Nova Scotia, Canada. POPULATION: All 126 604 nonanomalous, singleton deliveries >/=37 weeks in cephalic presentation from 1988-2002. METHODS: An analysis of maternal, antenatal, intrapartum, and neonatal factors associated with respiratory depression at birth >/=37 weeks. MAIN OUTCOME MEASURES: A composite outcome of delay in initiating and maintaining respiration after birth, 5-minute Apgar score

Measurement of Campylobacter numbers on carcasses in British poultry slaughterhouses.

An assessment of the proposed new International Organization for Standardization quantitative method for Campylobacter was undertaken on poultry carcass samples collected after the chilling phase of processing. Using a critical differences method, we determined the uncertainty associated with log-transformed Campylobacter numbers by dual analyses of 346 samples collected from 22 processing plants located throughout the United Kingdom. Overall, using log-transformed Campylobacter numbers that ranged between -1 and 5 log, we calculated the expanded measurement of uncertainty (EMU) to be 3.889 for the new method. The EMU changed when ranges of bacterial numbers were grouped for analyses. For low numbers of Campylobacter (< 1 log), the EMU was calculated to be 5.622. There was less measurement error with higher bacterial numbers because the EMU was found to be 0.612 for samples containing Campylobacter numbers of 3 log or above. The draft method was used to measure numbers of Campylobacters on poultry carcasses collected from 18 United Kingdom processing plants in summer and winter. Numbers were significantly lower in winter. We conclude that, although the new method is adequate at quantifying high numbers of Campylobacter on poultry carcasses, further development is required to improve the measurement of small numbers of this causative agent of foodborne illness.