Campylobacter Counts Research Articles

Usefulness of indicator bacteria as potential marker of Campylobacter contamination in broiler carcasses

According to Regulation (EC) No 2017/1495, amending Regulation (EC) No 2073/2005, a slaughterhouse process hygiene criterion based on a limit of 103 CFU/g of Campylobacter for no more than 20 (for the years 2018 and 2019), 15 (for the years 2020–2024) and 10 (starting from 2025) of 50 neck skin samples of broiler carcasses could be an effective measure to reduce the incidence of human campylobacteriosis. In order to stimulate the poultry industry to improve the control of Campylobacter along the slaughter-line, the quantification of indicator bacteria such as Escherichia coli or Enterobacteriaceae could be a useful strategy. The aims of this study were: a) to investigate the possible relationship between Campylobacter and indicator bacteria counts at two different points of the broiler slaughter-line and b) to evaluate the probability that carcasses have Campylobacter counts above 103 CFU/g in relation to indicator bacteria counts. E. coli, Enterobacteriaceae and Campylobacter were simultaneously enumerated on neck skin samples of broiler carcasses sampled at the post-evisceration (n = 75) and at the post-chilling points (n = 75) of three Italian poultry slaughterhouses. In general, the log counts of all the investigated microorganisms were significantly lower at the post-chilling point, and the indicator bacteria (E. coli and Enterobacteriaceae) counts were significantly higher than Campylobacter counts at both sampling points. A multilevel linear mixed model, relating the Campylobacter log10 counts with the E. coli and Enterobacteriaceae log10 counts, showed that the Campylobacter log10 counts increased significantly for every additional E. coli log10 count, and this was more evident at the post-chilling than at the post-evisceration sampling point. With regards to the Enterobacteriaceae, the increase was similar at the two sampling points. An additional model, developed to assess the probability that carcasses would have Campylobacter counts above 3 log10 CFU/g, showed that this probability increased significantly if the level of E. coli count also increased. Carcasses classified as Campylobacter > 3 log10 CFU/g according to the observed results of E. coli ≥ 4 log10 CFU/g did have high Campylobacter counts. Conversely, it was not possible to conclude anything for carcasses with E. coli < 4 log10 CFU/g. These findings support the hypothesis that the monitoring of poultry carcasses for E. coli load could be useful to identify those heavily contaminated with Campylobacter, in order to implement control measures on the farms of origin of such batches, or improving the slaughter process of the plants where the heavily contaminated batches are found.

Quantification of the Campylobacter contamination on broiler carcasses during the slaughter of Campylobacter positive flocks in semi-industrialized slaughterhouses

Campylobacter contamination of broiler carcasses has been little studied in semi-industrialized slaughterhouses in developing countries, where several steps are carried out manually or with limited technology. In this study, we performed quantification of the Campylobacter contamination on carcasses at four steps in the slaughter process in three Ecuadorian slaughterhouses. Therefore, 15 Campylobacter positive batches were sampled in three commercial slaughterhouses. For every batch, caecal content and five samples of breast skin were taken and examined for Campylobacter counts at the following steps: after plucking, after evisceration, after final washing and after water chilling. Slaughterhouse C was the only slaughterhouse in which Campylobacter counts increased significantly after evisceration. No significant differences were found between counts after evisceration and after final washing (P > 0.05). In all slaughterhouses, a significant reduction of Campylobacter counts (0.11 to 2.55 log10 CFU/g) was found after the chilling step. The presence of chlorine in the chilling water was associated with the highest reduction in Campylobacter counts on the carcasses. A high variability of Campylobacter counts was found within and between batches slaughtered in the same slaughterhouse. Campylobacter counts in caecal content samples were not correlated with counts on carcasses after plucking nor after evisceration.

The Prevalence of Campylobacter spp. in Polish Poultry Meat.

The prevalence, count and molecular identification of Campylobacter spp. in Polish poultry meat were analysed. 181 samples of meat from chicken (70), turkey (47), duck (54) and goose (10) were studied. Campylobacter spp. was found in 64% of meat samples. The highest prevalence of this pathogen was detected for duck meat. On average 80% of duck samples were contaminated with Campylobacter spp. The counts of Campylobacter spp. in positive samples remained under ten colony forming units per gram of product in 59% of poultry meat. C. jejuni was more frequently detected in poultry meat than C. coli.

Analysis of a poultry slaughter process: Influence of process stages on the microbiological contamination of broiler carcasses.

In a large-scale Swiss poultry abattoir, a microbiological process analysis of broiler carcasses was performed. At each selected process stage (scalding, plucking, evisceration, washing, and chilling), 90 carcasses from 30 flocks were sampled and examined for Campylobacter, Salmonella, Escherichia coli, Enterobacteriaceae, and extended-spectrum b-lactamases-producing Enterobacteriaceae. With regard to Campylobacter counts on carcasses, plucking tended to slightly increase the results (on average by 0.4 log CFU/g), whereas mean counts from plucked and chilled carcasses were comparable (3.1 log CFU/g after plucking, 3.0 log CFU/g in the chiller). The Campylobacter results of chilled carcasses are thereby likely to comply with the newly defined requirements of the European Union (process hygiene criterion for Campylobacter). With regard to Escherichia coli and Enterobacteriaceae counts on carcasses, plucking clearly reduced the results (on average by 0.8 and 0.9 log CFU/g), whereas mean counts from plucked and chilled carcasses were comparable (3.4 and 3.5 log CFU/g after plucking, 3.4 log CFU/g in the chiller). In contrast, Salmonella spp. were not detected on broiler carcasses and extended-spectrum b-lactamases- producing Enterobacteriaceae only rarely (1.8%). Such abattoir-specific data are of central importance for assessment of slaughter process performance and if necessary for the implementation of effective measures in the slaughter process.

β-Resorcylic Acid, a Phytophenolic Compound, Reduces Campylobacter jejuni in Postharvest Poultry

Human Campylobacter infections, a leading foodborne illness globally, has been linked with the high prevalence of this bacterium on raw retail chicken products. Reduction of Campylobacter counts on poultry products would greatly reduce the risk of subsequent infections in humans. To this end, this study investigated the potential of the phytophenolic compound β-resorcylic acid (BR) to reduce Campylobacter counts on postharvest poultry (chicken skin or meat). Four trials in total, two each on thigh skin or breast meat, were conducted in which chicken skin or meat samples (2 ± 0.1 g; 10 samples per treatment) were inoculated with 50 μL (∼106 CFU per sample) of a cocktail of four wild strains of C. jejuni. After 30 min of attachment, inoculated samples were dipped in a 0, 0.5, 1, or 2% BR solution for 30 s immediately followed by vigorously vortexing the samples in Butterfield's phosphate diluent and plating the supernatant for Campylobacter enumeration. In addition, the effect of BR on the color of skin and meat samples was studied. Moreover, the change in the expression of survival and virulence genes of C. jejuni exposed to BR was evaluated. Data were analyzed by the PROC MIXED procedure of SAS (P < 0.05; SAS Institute Inc., Cary, NC). All BR treatments significantly reduced Campylobacter populations on both chicken or meat samples by 1 to 3 log CFU/g compared with non-BR-treated washed controls. No significant difference in the lightness, redness, and yellowness of skin and meat samples was observed on exposure to BR wash (P > 0.05). Real-time PCR results revealed that BR treatment down-regulated expression of select genes coding for motility (motA, motB) and attachment (cadF, ciaB) in the majority of C. jejuni strains. Stress response genes (sodB, katA) were upregulated in C. jejuni S-8 (P < 0.05). Overall, our results suggest that BR could be effectively used as antimicrobial dip treatment during poultry processing for reducing Campylobacter on chicken carcasses.

Influence of different protein sources in the broiler diet on the presence of Campylobacter spp. in excreta and caecal content.

The aim of the present study was to examine possible effects of a different protein supply on the presence of Campylobacter in excreta and caecal content of broiler chickens. Therefore, 220 one-day-old chickens were fed five different diets in the experimental period (day 8 onwards). In the control group (CON; n=3 boxes), a soya bean-based industrially produced complete diet containing 5-10% whole wheat and coccidiostats was fed, whereas in the remaining groups, four own-made complete diets with four protein sources and combinations (4×2 boxes; soya bean meal-based diet=SBM; rapeseed meal-containing diet=RSM; haemoglobin meal-containing diet=HGP; algae-containing diet=ALG) were offered. During the 6-week trial period, data concerning performance parameters, excreta and litter characteristics and microbiological analysis of excreta and caecal content at dissection were collected. The qualitative bacteriological investigation was based on the EN ISO 10272-1:2006, taken from the official collection of analysis methods in accordance with § 64 LFBG. For quantitative bacteriological tests, a 10-fold dilution series was made followed by culture-based quantification methods on approved selective media in duplicate. Comparing the results of the final dissection (day 44/45), nearly all samples of animals receiving the own-made complete diet were Campylobacter positive in the caecal content, whereas only seven of fifteen samples were positive in the CON group. With regard to the CFU in the caecal content, the counts of Campylobacter were significantly lower for animals fed the commercial diet (2.47±3.06 vs. 7.36±0.66 log10 CFU/g). When only the positive animals were evaluated, significant differences were also found between the CON group and the total of the other groups (5.30±2.08 vs. 7.54±1.70 log10 CFU/g). Although significant differences were seen in the results, no final recommendation can be given how to create a diet to reduce an infection with Campylobacter.

Treatment with a low pH processing aid to reduce Campylobacter counts on broiler parts

New regulations and performance standards for Campylobacter have been implemented by the USDA - Food Safety and Inspection Service (FSIS). The objective of this study was to evaluate treatment with a low pH processing aid (CMS1 PoultrypHresh), a formulated low pH processing aid, to reduce numbers of Campylobacter which could help companies meet regulatory requirements. Two experiments (3 replicates each) were conducted. Experiment 1, in each of 3 replicates, skin-on split chicken breasts (n = 15) were obtained from a local grocery and divided into groups of 5. The skin of each part was inoculated with approximately 107 cells of a gentamicin resistant C. coli (CCGR) marker strain in an area of approximately 6.5 cm2. CCGR cells were allowed to attach for 5 min prior to treatment. Ten inoculated breasts were individually placed into separate 6 L plastic storage boxes containing either 3.5 L deionized water or PoultrypHresh solution at a pH of 1.4. Parts were subjected to agitation (bubbled air) for 25 s. After treatment, each part was removed, allowed to drain for 5 s, and placed into a plastic bag prior to mechanical rinsing with 150 mL of buffered peptone water for 60 s. Five inoculated breasts served as controls, were untreated with a dip or agitation and sampled as above. Experiment 2 procedures were repeated using skin-on thighs under the same conditions. Rinsates were collected from each chicken part, serially diluted, and plated onto Campy Cefex agar with 200 ppm gentamicin (CCGen). All plates were incubated microaerobically (5% O2, 10% CO2, 85% N2) for 48 h at 42°C, colonies were counted and the cfu/mL was log transformed. The use of PoultrypHresh on split breast produced a 99.6% reduction compared to untreated controls, while thighs showed a 99.4% reduction. This study demonstrated an approximate 3 log reduction (P < 0.05) using a 25 s air agitation treatment in PoultrypHresh at pH 1.4 with no observable damage, which will help processors meet FSIS regulations.

Monitoring Salmonella, Campylobacter, Escherichia coli and Staphylococcus aureus in traditional free-range 'Label Rouge' broiler production: a 23-year survey programme.

'Label Rouge' broiler free-range carcasses have been monitored since 1991, and broiler flocks since 2010, for contamination by the main foodborne zoonotic bacteria. Initially, the monitoring plan mainly focused on the surveillance of Salmonella, and on indicators of the overall microbiological quality of free-range broiler carcasses such as Staphylococcus aureus and coliforms, but was extended in 2007 to include Campylobacter enumeration on carcasses and in 2010, to Salmonella in the environment of live birds. Salmonella contamination of free-range broiler carcasses rose to a peak of 16% in 1994 but less than 1% of carcasses are now regularly found to be positive. Indicators of the overall microbiological quality of carcasses are also improving. These results correlate with the low prevalence of Salmonella in free-range broiler breeding and production flocks, and with the continuous improvement of hazard analysis and critical control points in slaughterhouses, the implementation of a good manufacturing practice guide since 1997 and the application of EU regulations on Salmonella since 1998 in France. Regarding Campylobacter counts on carcasses, the situation has been improving continuously over the last few years, even if 2·5% of the carcasses are still contaminated by more than 1000 Campylobacter per g of skin. Although the current control system focusing on Salmonella is based on firm epidemiologic data and offers effective means of control (e.g. slaughtering of positive breeder flocks), existing information on Campylobacter makes it more difficult to formulate an effective control plan for free-range broilers, due to their particular exposure to environmental contamination. This long-term surveillance programme provided an extended view of the evolution of the contamination of free-range broilers and a direct measurement of the impact of mandatory and profession-driven interventions on the microbiological quality of carcasses.

Impact of direct and indirect heating systems in broiler units on environmental conditions and flock performance

This study compared the impact of three indirect heating systems to direct gas flame heaters on a selection of flock performance and environmental indicators in commercial broiler units. No statistically significant differences (P≥0.05) were found in flock mortality rates, bird weight, water consumption, stress response, carbon dioxide, ammonia, temperature, relative humidity, litter quality, within-flock Campylobacter levels or mean Campylobacter counts when flock data from any of the three indirect heating systems were compared to flocks reared in houses with direct heating systems. Differences in litter quality were observed between upper and lower litter layers in all houses, regardless of heating type, which may have implications for bird health and welfare. Carbon dioxide concentrations in houses with direct heating systems were significantly higher than those in houses with indirect heating systems during the first 10 days of bird life (P≤0.05). This was due to the increased use of heating systems during this period of the flock cycle. Differences in CO2 concentrations had no effect on flock performance, possibly due to the fact that concentrations did not exceed known safe levels. A statistically significant increase in stress response was observed in birds as a result of partial depopulation (thinning) within houses, irrespective of heating system type used (P≤0.05). Stress associated with thinning may have consequences for bird welfare and food safety. In conclusion, the results of our study suggest that indirect heating systems do not appear to negatively impact on flock performance, stress response, within-flock Campylobacter levels or mean Campylobacter counts and do not appear to significantly alter environmental conditions within broiler houses when compared to houses equipped with direct heating systems. Indirect systems are a viable alternative for heating broiler houses in terms of flock performance, bird welfare and food safety.

TLR4 and TLR21 expression, MIF, IFN-β, MD-2, CD14 activation, and sIgA production in chickens administered with EFAL41 strain challenged with Campylobacter jejuni.

The protective effect of Enterococcus faecium EFAL41 on chicken's caecum in relation to the TLR (TLR4 and TLR21) activation and production of luminal IgA challenged with Campylobacter jejuni CCM6191 was assessed. The activation of MIF, IFN-β, MD-2 and CD14 was followed-up after bacterial infection. Day-old chicks (40) were divided into four groups (n=10): control (C), E. faecium AL41 (EFAL41), C. jejuni (CJ) and combined E. faecium AL41+C. jejuni (EFAL41+CJ). Relative mRNA expression of TLR4, TLR21 and CD14 was upregulated in the probiotic strain and infected (combined) group on day 4 and 7 post infection (p.i.). The caecal relative MD-2 mRNA expression was upregulated on day 4 p.i. in the EFAL41+CJ and CJ groups. MIF and IFN-β reached the highest levels in the combined groups on day 7 p.i. The concentration of the sIgA in intestinal flush was upregulated in EFAL41+CJ group on day 4 p.i. The results demonstrated that E. faecium EFAL41 probiotic strain can modulate the TLRs expression and modify the activation of MIF, IFN-β, MD-2 and CD14 molecules in the chickens caecum challenged with C. jejuni CCM 6191. The counts of EFAL41 were sufficient and high, similarly the counts of enterococci in both, caecum and faeces but without reduction of Campylobacter counts.

Campylobacter spp. and Escherichia coli contamination of broiler carcasses across the slaughter line in Danish slaughterhouses

This study presents levels of Campylobacter spp. and Escherichia coli on broiler carcasses across the slaughter line in three fully automated Danish slaughterhouses with the aim to investigate differences in slaughter hygiene between the lines, correlation between concentrations of E.coli and Campylobacter, and finally, the relationship between Campylobacter counts in caeca and on chilled meat. In total, 15 commercial, indoor flocks were examined and from each flock 24 caecal samples and 24 carcass samples were collected from each of the control points after plucking (AP), after evisceration (AE) and after chilling (AC). Results showed distinct differences between slaughterhouses. For slaughterhouse I the contamination level was high AP and decreased AE while for slaughterhouse II the contamination level was low AP and increased AE. For slaughterhouse III the contamination level varied insignificantly across the processes. Results also showed differences in contamination levels of E. coli and Campylobacter between slaughterhouses. Mean counts of the two organisms increased or decreased concurrently from after plucking to after evisceration within slaughterhouses; however, after chilling counts of E. coli were reduced to a larger extent than counts of Campylobacter. This suggests that for processing of Campylobacter positive broilers E. coli may be used as an indicator of faecal contamination during the processing steps up to the point of chilling but not as an indicator of Campylobacter contamination of chilled broiler meat. A correlation was found, though, between the mean number of Campylobacter in caeca and the mean number of Campylobacter on broiler meat after chilling which means that the level in the gut at slaughter significantly impacts the level on the chilled meat. In conclusion, our data confirm that less faecal contamination throughout processing, and/or less Campylobacter in the gut at the point of slaughter will lead to less Campylobacter contamination on the meat and thereby improve food safety. Exchange of information between slaughterhouses on best hygiene practices and compliance with these is an option to reduce numbers of Campylobacter in broiler meat.

Application of TRiMiCri for the evaluation of risk based microbiological criteria for Campylobacter on broiler meat

A potential solution for the reduction of consumer exposure to Campylobacter is establishing a microbiological criterion (MC) for Campylobacter on broiler meat. In the present study the freely available software tool TRiMiCri was applied to evaluate risk-based microbiological criteria by two approaches: the traditional one that implies a microbiological limit (ML-MC) and the second one which is based on the relative risk estimate (RRL-MC). A baseline risk was estimated based on the Belgian baseline data, whereas the data for the evaluated batches were Campylobacter counts from 28 Campylobacter positive batches sampled in six Belgian slaughterhouses. Results showed that approximately 60% of Campylobacter positive batches did not comply with ML criteria based on the n=5, m=1000 and c=0 for ML-MC and equivalently for RRL criteria when RRcrit=1. As expected, the less stringent MCs decreased the percentages of non-compliance (NC) but they are less effective, as reflected in increased minimum relative residual risks (MRRRs). When the Belgian baseline is used, more batches are found to be compliant than when the Danish baseline provided by TRiMiCri is used. Based on this, the application of microbiological criteria for Campylobacter in the EU is discussed. TRiMiCri provides user friendly software to evaluate the available data and can help risk managers in establishing risk based microbiological criteria for Campylobacter in broiler meat.

Efficacy of feed additives against Campylobacter in live broilers during the entire rearing period: Part B

A total of 636 day-of-hatch Ross 308 broilers chicks were used in 4 independent trials carried out to screen the effect of 12 feed additives on reducing cecal colonization of Campylobacterin broilers. The tested additives were probiotics based on B. subtilis and S. cerevisae, a garlic extract, a blend of herbal substances and essential oils, two different combinations of essential oils and organic acids (OA), two mixtures of flavoring compounds, medium chain fatty acids (MCFA), monoglycerides (MG) of MCFA and MG-MCFA+OA. At 14 days of age, all the birds were orally infected with 0.1 mL of a bacterial suspension of C. jejuni ST-45 diluted at 10(5) cfu/mL in tryptone salt broth. In each trial, there was a positive control group and 2 (Trials 1 and 2) or 4 (Trials 3 and 4) additional treatment groups supplemented with additives, which were added to feed or water only to the finisher (21 to 42 d) diet (Trials 1 and 2) or to the starter (0 to 21 d) and finisher diets (Trials 3 and 4). Feed and water were available ad libitum. On days 35 and 42 of age in Trials 1 and 2, and on days 21, 35 and 42 of age in Trials 3 and 4, 10 (Trials 1 and 2) or 12 birds (Trials 3 and 4) per group were euthanized for cecal sampling. In Trial 1, birds fed with MCFA and MG-MCFA had a significant (P<0.05) reduction in cecal Campylobacter colonization compared to control at 35 d, but only the group treated with MG-MCFA maintained the reduction at 42 d. In Trials 2 to 4, no significant differences (P>0.05) in cecal Campylobacter counts were found between the treated and control animals. In conclusion, although none of the treatments were able to completely prevent the colonization of chickens with C. jejuni, MCFA and MG-MCFAs could reduce the pathogen counts when supplemented from 21 days onwards.

Identification of risk factors for Campylobacter contamination levels on broiler carcasses during the slaughter process

Campylobacter carcass contamination was quantified across the slaughter line during processing of Campylobacter positive batches. These quantitative data were combined together with information describing slaughterhouse and batch related characteristics in order to identify risk factors for Campylobacter contamination levels on broiler carcasses. The results revealed that Campylobacter counts are influenced by the contamination of incoming birds (both the initial external carcass contamination and the colonization level of caeca) and the duration of transport and holding time that can be linked with feed withdrawal period. In addition, technical aspects of the slaughter process such as a dump based unloading system, electrical stunning, lower scalding temperature, incorrect setting of plucking, vent cutter and evisceration machines were identified as risk factors associated with increased Campylobacter counts on processed carcasses. As such the study indicates possible improvements of the slaughter process that can result in better control of Campylobacter numbers under routine processing of Campylobacter positive batches without use of chemical or physical decontamination. Moreover, all investigated factors were existing variations of the routine processing practises and therefore proposed interventions are practically and economically achievable.

The Effects of Sulfadimethoxine Administered to Control Campylobacter jejuni in Small-Scale Broiler Operations

Campylobacter jejuni causes human foodborne gastroenteritis known as campylobacteriosis. Antimicrobial therapy could be a potentially important tool in reducing the prevalence of C. jejuni in poultry. The purpose of this study was to determine the effects of sulfadimethoxine antibiotic on the prevalence of C. jejuni in growing broilers. Day-old broilers (n= 600) were allotted to two treatments 1) control (drinking water only) and 2) antibiotic (drinking water + 0.05% (wt/vol) sulfadimethoxine) with two replications. Each week, fecal samples were collected from individual chickens (n=300). All samples were plated on modified charcoal-cefoperazone-deoxycholate agar (mCCDA) to determine the log CFU/g and prevalence (%) of Campylobacter spp. Isolation of C. jejuni was verified with latex agglutination and hippurate hydrolysis test. Over the six week period, the bacterial counts of Campylobacter spp. in the antibiotic treatment (5.12 log CFU/broiler) were significantly lower (P<0.05) than in the control treatment (6.05 log CFU/broiler). Additionally, the prevalence of C. jejuni in the antibiotic treatment (50.0%) was significantly lower (P<0.05) than in the control treatment (56.0%). Our findings suggest that the antibiotic sulfadimethoxine may aid in reducing Campylobacter spp. and the prevalence on both Campylobacter spp. and C. jejuni in growing broilers.

Superoxide dismutase SodB is a protective antigen against Campylobacter jejuni colonisation in chickens

Campylobacter is the leading cause of foodborne diarrhoeal illness in the developed world and consumption or handling of contaminated poultry meat is the principal source of infection. Strategies to control Campylobacter in broilers prior to slaughter are urgently required and are predicted to limit the incidence of human campylobacteriosis. Towards this aim, a purified recombinant subunit vaccine based on the superoxide dismutase (SodB) protein of C. jejuni M1 was developed and tested in White Leghorn birds. Birds were vaccinated on the day of hatch and 14 days later with SodB fused to glutathione S-transferase (GST) or purified GST alone. Birds were challenged with C. jejuni M1 at 28 days of age and caecal Campylobacter counts determined at weekly intervals. Across three independent trials, the vaccine induced a statistically significant 1 log10 reduction in caecal Campylobacter numbers in vaccinated birds compared to age-matched GST-vaccinated controls. Significant induction of antigen-specific serum IgY was detected in all vaccinated birds, however the magnitude and timing of SodB-specific IgY did not correlate with lower numbers of C. jejuni. Antibodies from SodB-vaccinated chickens detected the protein in the periplasm and not membrane fractions or on the bacterial surface, suggesting that the protection observed may not be strictly antibody-mediated. SodB may be useful as a constituent of vaccines for control of C. jejuni infection in broiler birds, however modest protection was observed late relative to the life of broiler birds and further studies are required to potentiate the magnitude and timing of protection.

The efficacy of selected probiotic and prebiotic combinations in reducing Campylobacter colonization in broiler chickens

Campylobacter is one of the leading causes of foodborne illness worldwide and is associated with consumption of poultry products. One potential strategy to reduce Campylobacter colonization in poultry is the use of probiotics. In an attempt to reduce Campylobacter in poultry, we isolated 117 GRAS (generally recognized as safe) bacterial isolates from the ceca of healthy birds and evaluated their efficacy against Campylobacter, using a soft agar overlay technique. Twenty-six isolates (Bacillus and Lactobacillus spp.) with greatest ability to inhibit Campylobacter in vitro, were tested in birds. In these studies, bacterial isolates were administered orally on day-of-hatch and, chicks were challenged with a four-strain mixture of wild type C. jejuni (∼106 CFU/mL) on day 7. Cecal samples were collected on day 14 for Campylobacter enumeration. Out of 26 isolates, 3 isolates demonstrated a reduction in Campylobacter counts (1 to 2 logs). In an attempt to improve the efficacy of these 3 isolates, follow-up studies were conducted with various doses of 2 prebiotics (fructooligosaccharide [FOS] or mannan oligosaccharide [MOS]) in the feed. Campylobacter counts were not reduced for any of the FOS treatments (0.125%, 0.25%, or 0.5%) or the MOS treatments (0.04%, 0.08%, or 0.16%), but were reduced for the combination of isolate 3 with 0.04% MOS in feed. The results of this study indicate that the selection and administration of bacterial isolates in combination with selected prebiotics may reduce enteric Campylobacter counts in preharvest poultry.

Use of Caprylic Acid in Broiler Chickens: Effect on Campylobacter jejuni.

The effect of caprylic acid (CA) on Campylobacter jejuni in chickens was evaluated using two approaches: dietary supplementation or surface treatment of chilled chicken carcasses. To analyze the dietary effect of CA, individually housed broiler chickens (n = 48) were artificially infected with C. jejuni VFU612 (10(6) colony-forming units [CFU]/bird) on the 21st and 35th days of life. Dietary CA (2.5 and 5 g/kg of feed, fed throughout the entire experiment) significantly decreased C. jejuni shedding (p<0.05). However, the effect only lasted for 3-7 days after infection. The numbers of Campylobacter shed by the positive control birds reached its maximum on the 37th day of life, while on that same day, both Treatment I and Treatment II groups shed significantly lower (p<0.05) numbers of Campylobacter (by 0.8 and 1.8 log10 CFU/g, respectively). Also, peak shedding was delayed by 1 day in both treated groups. After euthanasia of each chicken on the 42nd day of life, no differences in Campylobacter counts in the crop, gizzard, ileum, and cecum were found between the positive control and the treated groups (p>0.05). Surface contamination of the chilled chicken halves was performed with C. jejuni VFU612 (clinical isolate) and CCM6214 (collection strain). Surface treatment with CA at 1.25 and 2.5 mg/mL for 1 min significantly reduced C. jejuni VFU612 contamination of chicken skin (p<0.05) by 0.29-0.53 and 1.14-1.58 log10 CFU/g of skin, respectively. Counts of C. jejuni CCM6214 were reduced by 0.68-1.65 log10 CFU/g of skin). In conclusion, dietary CA affected numbers of C. jejuni in the gastrointestinal contents of chickens, whereas surface treatment reduced C. jejuni contamination in processed chicken carcasses.

Assessment of microbial load in marketed broiler meat at Mymensingh district of Bangladesh and its public health implications

An investigation was conducted to assess the bacteriological quality of 30 samples of fresh broiler thigh meat samples sold in different retail markets in Mymensingh. Total viable count (TVC), total coliform count (TCC), total salmonella count (TSC) and total campylobacter count (TCpC) in meat samples of different broiler markets like K.R. at BAU campus, Boyra and Kewatkhali were determined. Mean of TVC, TCC, TSC and TCpC for the K.R. at BAU campus, Boyra and Kewatkhali markets were 5.69, 6.03, 6.17 log10 CFU/g, 4.52, 4.66, 4.69 log10 CFU/g, 3.35, 3.51, 3.61, log10 CFU/g and 2.31, 2.56, 2.66 log10 CFU/g, respectively. It was observed that the mean values of TVC, TCC, TSC and TCpC in case of Boyra and Kewatkhali market exceeded the ICMSF recommendations which may cause alarm to consumer’s health. The variation of TVC in meats of different broiler market was significant (P&lt;0.05) at 5% level of probability whereas TCC and TSC obtained from meat samples of different markets were not showed significant (P&gt;0.05).The mean values of TCpC in meats of three different market were highly significant with 1% level of probability (P&lt;0.01).There was no significant correlation found between TVC and TCC (P&gt;0.05), but a significant correlation found between TVC with TSC and TVC with TCpC in meats of three different markets respectively. Presence of Escherichia coli, Campylobacter and Salmonella spp. in meats must receive particular attention, as these organisms are responsible for causing harm to public health. Suggestion has been given to improve the present sanitary condition of meat processing to minimize bacterial load.Res. Agric., Livest. Fish.2(1): 87-95, April 2015

Intracloacal Inoculation, an Effective Screening Method for Determining the Efficacy of Probiotic Bacterial Isolates against Campylobacter Colonization in Broiler Chickens

Campylobacter is a leading cause of foodborne illness worldwide. It is common in poultry, and human infections are often associated with consumption of contaminated poultry products. One strategy to reduce Campylobacter colonization in poultry is the use of oral probiotics, but this produces variable results, possibly because the probiotics are destroyed in the stomach's acidic environment. Protection (e.g., encapsulation) of isolates may overcome this problem, but there is no assurance that these isolates will have efficacy in the lower gastrointestinal tract. Therefore, screening candidate isolates by directly placing them in the lower intestinal tract via cloacal inoculation may eliminate the time and expense of encapsulating ineffective isolates. Thus, the purpose of this study was to collect bacterial isolates with anti-Campylobacter activity in vitro and evaluate their efficacy in vivo upon either oral or intracloacal administration. Bacterial isolates were collected from healthy birds and were evaluated for efficacy against C. jejuni in vitro. Isolates having generally regarded as safe status and demonstrating in vitro anti-Campylobacter properties were evaluated after oral or intracloacal inoculation into chicks on day 1 (n = 10 birds per isolate per route of administration). On day 7, birds were dosed by oral gavage with a four-strain mixture of wild-type Campylobacter containing at least 1 x 107 CFU/ml organisms. On day 14, birds were euthanized and the ceca were collected aseptically for Campylobacter enumeration. When dosed orally, only one isolate had a 1-log reduction in cecal Campylobacter counts, whereas when administered intracloacally, six of these isolates produced a 1- to 3-log reduction in cecal Campylobacter counts in 14-day-old chickens. These results support the strategy of evaluating the efficacy of potential probiotic isolates via cloacal inoculation prior to undergoing the effort of encapsulating isolates for oral administration.