Campylobacter Concentrations Research Articles

Detection probability of Campylobacter

A rapid presence/absence test for Campylobacter in chicken faeces is being evaluated to support the scheduling of highly contaminated broiler flocks as a measure to reduce public health risks [Nauta, M. J., & Havelaar, A. H. (2008). Risk-based standards for Campylobacter in the broiler meat chain. Food Control, 19, 372–381]. Although the presence/absence test is still under development, an example data set of test results is analysed to illustrate the benefit of the detection probability concept. The detection probability of Campylobacter increases with the logarithm of the Campylobacter concentration in faeces according to an S-shaped curve which stretches about 2–3 log units. The detection probability is 50% at a Campylobacter concentration of 7.4 × 10 6 cfu/g. The uncertainty in the detection probability is 32% at the most for a 90% confidence interval. This type of information allows for realistic calculations on the Campylobacter status of different food processing paths after splitting. Usable quantitative estimates on detection probability await a data set of test results from a test that is ready for use or has similar properties.

Campylobacter biofilm phenotype exhibits reduced colonization potential in young chickens and altered in vitro virulence

In this study, we evaluated the ability of different Campylobacter phenotypes (biofilm versus planktonic) to colonize young poultry. It has been suggested that a persistent Campylobacter biofilm reservoir may be involved in the initial contamination of poultry flocks. Campylobacter jejuni cultured adherent to agar was utilized as the biofilm model and C. jejuni cultured in broth was evaluated as the planktonic model. In 2 independent trials, 1-d-old broiler chicks were given 1 of 3 treatments: 1) 10(5) cfu.mL(-1) of C. jejuni cultured in broth, 2) 10(5) cfu.mL(-1) of C. jejuni cultured adherent to agar, or 3) no C. jejuni (negative control). Cecal contents of all birds were evaluated by culturing 12 d after the initial challenge with C. jejuni. In both trials, birds challenged with C. jejuni cultured in broth had approximately 3 to 4 log higher cecal Campylobacter concentration than birds challenged with C. jejuni cultured adherent to agar. Using 2 cell lines (INT 407 and DF1), virulence of C. jejuni cultured in broth versus adherent to agar also was evaluated by challenging monolayers of eukaryotic cells with 1 of 3 treatments: 1) 10(5) cfu.mL(-1) of C. jejuni cultured in broth, 2) 10(5) cfu.mL(-1) of C. jejuni cultured adherent to agar, or 3) no C. jejuni (negative control). The virulence study also showed differences of C. jejuni cultured in broth or agar in attachment and invasion abilities to tissue culture cells, but differences were not as consistent as with the chick colonization study. This study indicates that phenotype may play a role in colonization of chickens and virulence by C. jejuni.

Reduction of thermotolerant Campylobacter species on broiler carcasses following physical decontamination at slaughter.

To reduce the incidences of human Campylobacter infections, a number of countries are investigating methods for reducing human exposure to Campylobacter from broiler meat. In addition to implementing biosecurity measures at the farm, Campylobacter may be controlled by reducing Campylobacter counts through physical decontamination of the meat. The current study was conducted to compare the Campylobacter-reducing ability of three physical decontamination techniques, forced air chilling, crust freezing, and steam-ultrasound, performed in the plant with naturally contaminated broiler chickens. The effects of all three techniques were evaluated and compared with the effect of freezing. Mean reductions obtained were 0.44 log CFU per carcass, 0.42 log CFU per sample, and > or = 2.51 log CFU per carcass, respectively. All techniques resulted in significant reductions of the Campylobacter concentration on the carcasses (P < 0.05). However, none of the techniques were as effective as freezing based on reductions in Campylobacter counts and on adverse effects. The increase in Campylobacter counts on carcasses following visceral rupture during the evisceration operation also was examined. Visceral rupture resulted in an increase of 0.9 log CFU per carcass, suggesting that Campylobacter counts also may be reduced by optimizing the hygienic design of equipment or by physical removal of fecal contamination.

Purified Cell Wall of Saccharomyces cerevisiae Increases Protection Against Intestinal Pathogens in Broiler Chickens

A study was conducted to determine effects of a mannanoligosaccharide prebiotic, derived from cell wall of the yeast Saccharomyces cerevisiae, on morphological development of the intestines and microbial populations of the ceca and litter. Dietary treatments included: antibiotic-free diet (CTL), diet 1 + virginiamycin (VIRG; 16.5 mg/kg feed) and diet 1 + ActiveMOS (MOS; 1.5 kg/T starter diet and 1 kg/T grower diet). Each treatment was assigned to 3 pen replicates (55 birds/pen). At day 14, 24 and 34, cecal contents were used for Lactobacilli, Bifidobacteria, E. coli and Campylobacter quantification whereas litter was analyzed for Campylobacter and E. coli. At same time points, jejunum samples were used in histological analysis. MOS significantly increased goblet cell number in the jejunum (p<0.05) at day 24 and 34. In contrast to the CTL and VIRG diet, MOS consistently increased cecal populations of Bifidobacteria (p<0.05) at all times. Moreover, at day 34, MOS increased cecal populations of Lactobacilli (p<0.05) and reduced E. coli and Campylobacter concentrations (p<0.05). None of the dietary treatments altered E. coli and Campylobacter concentrations in the litter. In comparison to antibiotics, MOS, therefore, improved intestinal health conditions by increasing goblet cell number into the villi membrane, stimulating growth of beneficial bacteria and reducing colonization by pathogenic bacteria.

Microbial Recovery from Genetically Featherless Broiler Carcasses after Forced Cloacal Fecal Expulsion

A study was conducted to determine external microbiology of genetically featherless broiler carcasses after forced cloacal fecal expulsion. Full-fed featherless broilers were placed into coops, transported, unloaded, shackled, stunned, suffocated, weighed, and divided into 3 treatments groups. Carcasses were transferred to a separate shackle line and passed through a machine designed to induce defecation (squeeze) and then remove external feces (wash). Treatments were obtained by turning the squeezing and washing components on or off. Treatments were as follows: S carcasses were squeezed but not washed; W carcasses were not squeezed but were washed; and SW carcasses were squeezed and washed. Concentrations of total aerobic microorganisms (AB), Escherichia coli (EC), coliforms (CF), and Campylobacter (CPY) recovered from whole carcass rinses did not vary with treatment (P > 0.05). However, counts of Salmonella (SAL) in rinses of S carcasses were 1.4 log10 cfu/mL greater than counts of SAL found in rinses of SW carcasses (P < 0.05). The SAL prevalence was similar for S (86% positive), W (90% positive), and SW (83% positive) carcasses (P > 0.05). Populations of AB and CF recovered from wash water (water applied in the machine after fecal expulsion) for SW carcasses were significantly higher by 3.1 and 1.5 log10 cfu/mL, respectively, than the populations of the same bacteria recovered from wash water for W carcasses (P < 0.05). Levels of EC and CPY recovered from wash water did not vary with treatment. There was no difference in CPY and SAL prevalence in water collected after washing W carcasses or SW carcasses (P > 0.05). Data from the present study show that controlled cloacal fecal expulsion followed by carcass washing immediately after slaughter can be used to minimize the numbers of carcass Salmonella and can reduce the likelihood of visible carcass fecal contamination or cross-contamination to other carcasses and processing equipment.

Prevalence and Concentration ofCampylobacterin Rumen Contents and Feces in Pasture and Feedlot-Fed Cattle

Campylobacter are important human foodborne pathogens known to colonize the gastrointestinal tract of cattle. The incidence of Campylobacter in cattle may be seasonal and may vary among age groups and type (beef versus dairy). Less is known about other factors that could influence the prevalence, colonization site, and shedding of Campylobacter in cattle. The objectives of this study were to evaluate the prevalence and enumerate Campylobacter at two sites along the digestive tract of beef and dairy type cattle consuming either grass or feedlot diets. In an initial study, Campylobacter was not recovered from rumen samples of any of 10 ruminally cannulated (six dairy and four beef type) pasture-reared cattle and there was no difference (p > 0.05) between cattle types on fecal Campylobacter recovery, with 50% of each type yielding culture-positive feces (overall mean +/- SE, 0.75 +/- 0.001 SEM log(10) colony-forming units [CFU]/g feces). When calculated from Campylobacter culture-positive animals only, mean fecal concentrations were 1.50 +/- 0.001 SEM log(10) CFU/g. In a follow-up study with feedlot and pasture-reared cattle (n = 18 head each), 78% of rumen and 94% of fecal samples from pastured cattle were positive for Campylobacter while 50% of the rumen and 72% of the fecal samples were positive in concentrate-fed animals. Overall mean concentration of Campylobacter was greater in feces than ruminal fluid (p < 0.05). When only Campylobacter-positive animals were analyzed, concentrations recovered from feces were higher (p < 0.05) in concentrate-fed than in pasture-fed cattle (4.29 vs. 3.34 log(10) CFU/g, respectively; SEM = 0.29). Our results suggest that the rumen environment and its microbial population are less favorable for the growth of Campylobacter and that concentrate diets may provide a more hospitable lower gastrointestinal tract for Campylobacter.

Baseline data from a Belgium-wide survey of Campylobacter species contamination in chicken meat preparations and considerations for a reliable monitoring program.

From February to November 2007, chicken meat preparations (n = 656) were sampled at 11 processing companies across Belgium. All samples were tested for Campylobacter by enrichment culture and by direct plating according to standard methods. Almost half (48.02%) of the samples were positive for Campylobacter spp. The mean Campylobacter count was 1.68 log(10) CFU/g with a standard deviation of +/- 0.64 log(10) CFU/g. The study revealed a statistically significant variation in Campylobacter contamination levels between companies; processors with a wider frequency distribution range of Campylobacter counts provided chicken meat preparations with higher Campylobacter incidences and concentrations. There was no significant difference between the counts of Campylobacter spp. in various preparation types. However, the Campylobacter counts and incidences in chicken wings were the highest and portioned-form products (legs, wings, and breasts) showed a higher probability of being Campylobacter positive compared to minced-form products (sausages, burgers, and minced meat). The proportion of Campylobacter-positive samples was significantly higher in July than in other months. Recovery of Campylobacter spp. recovery by direct plating was higher (41.0%) compared to detection after enrichment (24.2%). Statistical modeling of the survey data showed that the likelihood of obtaining a positive result by enrichment culture increases with an increase in the Campylobacter concentration in the sample. In the present study, we provide the first enumeration data on Campylobacter contamination in Belgian chicken meat preparations and address proposals for improving Campylobacter monitoring programs.

Caprylic Acid Supplemented in Feed Reduces Enteric Campylobacter jejuni Colonization in Ten-Day-Old Broiler Chickens ,

Campylobacter is one of the leading causes of human foodborne illness in the United States, and epidemiological evidence indicates that poultry and poultry products are a significant source of human Campylobacter infections. Reducing Campylobacter in the intestinal tract would reduce contamination of poultry products and eggs. Caprylic acid, an 8-carbon medium-chain fatty acid has been shown to be bactericidal against several pathogenic bacteria. It has, however, not been tested in the control of Campylobacter in chickens. Four trials were carried out to evaluate the efficacy of caprylic acid against cecal Campylobacter jejuni colonization in 10-d-old chicks. In the first 2 trials, day-of-hatch chicks (n=40 per trial) were assigned to negative controls (no Campylobacter, no caprylic acid), positive controls (Campylobacter, no caprylic acid), and a low (0.7%) and a high (1.4%) dose of caprylic acid supplemented in regular chick starter feed (n=10 chicks/treatment). Two more trials were carried out to evaluate a wider range of caprylic acid doses on cecal Campylobacter counts, in which day-of-hatch chicks (n=90 per trial) were assigned to 9 treatments: negative controls (no Campylobacter, no caprylic acid) and caprylic acid doses of 0 (positive controls), 0.35, 0.525, 0.7, 0.875, 1.05, 1.225, and 1.4% (n=10 chicks/treatment). Except for the negative controls, chicks were orally gavaged with approximately 1 x 10(6) cfu Campylobacter on d 3. On d 10, cecal contents were collected and Campylobacter concentrations were determined in each trial. In all 4 trials, the 0.7% dose of caprylic acid consistently reduced Campylobacter content counts compared with the positive control. In trials 3 and 4, doses less than 1.05% consistently reduced cecal Campylobacter content in both trials. At the higher doses, caprylic acid reduced feed consumption and body weight, but did not affect feed conversion when compared with the positive controls. These data suggest that low-dose supplementation with caprylic acid in feed may reduce Campylobacter colonization in young chickens.

Summary of the Swedish Campylobacter Program in Broilers, 2001 through 2005

A Campylobacter monitoring program in broiler chickens was carried out in Sweden from 2001 through 2005. The objective was to reduce the occurrence of Campylobacter in the food chain through preventive measures, starting with primary production. The program involved collecting samples from all broiler flocks at slaughter and occasional additional times. The annual incidence of Campylobacter-positive slaughter batches progressively decreased from 20% in 2002 to 13% in 2005. Most of the positive batches had a high within-flock prevalence of Campylobacter. However, about 18% of the positive batches had a low-within-flock prevalence; Campylobacter spp. were isolated from at most 50% of the cloacal samples. The incidence of batches contaminated at slaughter ranged between 6 and 9% during the study period. During all 5 years, a seasonal peak of incidence was observed in the summertime. In an additional study, quantitative analyses were performed on neck skin samples and carcass rinse samples. Those results were compared with the positive and negative findings of the cloacal, cecum, and neck skin samples at slaughter. When Campylobacter was found in the cecum, there was a higher level of Campylobacter in the quantitative analyses. Those batches where Campylobacter already had been found on the farm had a higher concentration of Campylobacter than those batches in which Campylobacter was found only at slaughter. During the study period, about one-third of producers seldom delivered Campylobacter-positive batches (<10% positive batches per year). Thus, it is possible to produce Campylobacter-free broilers in Sweden.

A collaborative study on a Nordic standard protocol for detection and enumeration of thermotolerant Campylobacter in food (NMKL 119, 3. Ed., 2007)

A Nordic standard protocol for detection and enumeration of thermotolerant Campylobacter in food has been elaborated (NMKL 119, 3. Ed., 2007). Performance and precision characteristics of this protocol were evaluated in a collaborative study with participation of 14 laboratories from seven European countries. The laboratories performed qualitative, semi-quantitative and quantitative analyses on samples of chicken meat, cut lettuce, and milk artificially inoculated with different concentrations including blank duplicates of one strain of Campylobacter coli (SLV-271) and one strain of Campylobacter jejuni (SLV-542). Expected concentrations (95% C.I.) (cfu g − 1 or ml − 1 ) of both strains in matrices were 0.6–1.4 and 23–60 for qualitative detection, and 0.6–1.4; 23–60; and 420–1200 for semi-quantitative detection. For quantitative determination, the expected concentrations of C. jejuni/ C. coli were 420–1200/580–1100; 2100–6000/6300–11,000; and 4100–11,000/53,000–97,000 cfu g − 1 or ml − 1 . The qualitative and semi-quantitative techniques resulted in comparable detection. The overall specificity and sensitivity of the detection techniques was 98.6% (95% C.I.: 95.1–99.8%) and 82.8% (C.I.: 78.4–86.6%), respectively. The sensitivity was not influenced by food matrix ( P = 0.359), but was significantly lower for C. coli compared to C. jejuni ( P = 0.007) and for concentrations below 1.4 cfu g − 1 ( P < 0.001). The detection techniques were therefore only considered satisfactory for detection of Campylobacter concentrations above approximately 25 cfu g − 1 for all matrices tested, which was supported by calculation of values for accordance, concordance, and concordance odds ratio. No statistical difference was found between enumerations obtained by the semi-quantitative and quantitative techniques for comparable concentrations of Campylobacter (420–1200 cfu g − 1 or cfu ml − 1 ) ( P = 0.104). Both techniques underestimated concentrations of thermotolerant Campylobacter in milk. The semi-quantitative technique estimated low inoculation levels of Campylobacter more correctly than the high inoculation levels. Counts obtained on the two selective plating media, Abeyta–Hunt–Bark agar added to it 0.1% triphenyl tetrazolium chloride and modified charcoal cephoperazone desoxycholate agar were not significantly different ( P = 0.143). Expressed as an absolute difference between log 10-transformed results, the overall values for repeatability ( r) and reproducibility ( R) were r = log 10 0.43 and R = log 10 1.99, respectively. By omitting results from laboratories with high level of variability in results, R was reduced to log 10 1.88. We suggest that the poor detection of low numbers, the underestimation in milk samples, and the large variation between laboratories can be explained by the general difficulties in handling Campylobacter. In conclusion, NMKL 119, 3. Ed., 2007, is regarded as an acceptable protocol for detection of thermotolerant Campylobacter at concentrations above 25 cfu g − 1 and also for enumeration of thermotolerant Campylobacter in chicken meat.

Effects of Short‐Chain Nitrocompounds against Campylobacter jejuni and Campylobacter coli in vitro

Effects of 2-nitro-1-propanol, 2-nitroethanol, nitroethane, and 2-nitro-methyl-propionate (0, 10, and 20 mM) on growth of Campylobacter jejuni were tested during culture in Bolton broth adjusted to pH 5.6, 7.0, or 8.2. The nitrocompounds were similarly tested against C. coli but at pH 8.2 only. Viable cell counts measured during incubation revealed main effects (P < 0.05) of all nitrocompounds on the survivability of C. jejuni. An effect of pH (P < 0.05) on the survivability of C. Jejuni during incubation with nitrocompounds was observed, with greater inhibition observed at pH 8.2 than at pH 5.6 or 7.0 for nitroethane, 2-nitro-l-propanol, and 2-nitroethanol, but not for 2-nitro-methyl-propionate, which showed greatest inhibition at pH 5.6. Except for 2-nitro-methyl-propionate, which was ineffective, all nitrocompounds elicited similar effects on C. coli. The effect of nitroethane and 2-nitro-l-propanol (10 mM) on naturally occurring Campylobacter was investigated during incubation of porcine fecal suspensions, where Campylobacter concentrations decreased more rapidly (P < 0.05) in suspensions with added 2-nitro-l-propanol than in unsupplemented or nitroethane-supplemented suspensions, thus reiterating the superior inhibitory effect of 2-nitro-l-propanol.

Zoonotic bacterial populations, gut fermentation characteristics and methane production in feedlot steers during oral nitroethane treatment and after the feeding of an experimental chlorate product

Nitroethane inhibits the growth of certain zoonotic pathogens such as Campylobacter and Salmonella spp., foodborne pathogens estimated to cause millions of human infections each year, and enhances the Salmonella- and Escherichia coli-killing effect of an experimental chlorate product being developed as a feed additive to kill these bacteria immediately pre-harvest. Limited studies have shown that nitroethane inhibits ruminal methane production, which represents a loss of 2–12% of the host's gross energy intake and contributes to global warming and destruction of the ozone layer. The present study was conducted to assess the effects of 14-day oral nitroethane administration, 0 (0X), 80 (1X) or 160 (2X) mg nitroethane/kg body weight per day on ruminal and fecal E. coli and Campylobacter, ruminal and fecal methane-producing and nitroethane-reducing activity, whole animal methane emissions, and ruminal and fecal fermentation balance in Holstein steers ( n = 6 per treatment) averaging 403±26 (SD) kg BW. An experimental chlorate product was fed the day following the last nitroethane administration to determine effects on E. coli and Campylobacter. The experimental chlorate product decreased ( P < 0.001 ) fecal, but not ruminal ( P > 0.05 ) E. coli concentrations by 1000- and 10-fold by 24 and 48 h, respectively, after chlorate feeding when compared to pre-treatment concentrations (>5.7 log 10 colony forming units/g). No effects ( P > 0.05 ) of nitroethane or the experimental chlorate product were observed on fecal Campylobacter concentrations; Campylobacter were not recovered from ruminal contents. Nitroethane treatment decreased ( P < 0.01 ) ruminal (8.46, 7.91 and 4.74±0.78 μmol/g/h) and fecal (3.90, 1.36 and 1.38±0.50 μmol/g/h) methane-producing activity for treatments 0X, 1X and 2X, respectively. Administration of nitroethane increased ( P < 0.001 ) nitroethane-reducing activity in ruminal, but not fecal samples. Day of study affected ruminal ( P < 0.0001 ) but not fecal ( P > 0.05 ) methane-producing and nitroethane-reducing activities ( P < 0.01 ); treatment by day interactions were not observed ( P > 0.05 ). Ruminal accumulations of acetate decreased ( P < 0.05 ) in 2X-treated steers when compared with 0X- and 1X-treated steers, but no effect ( P > 0.05 ) of nitroethane was observed on propionate, butyrate or the acetate to propionate ratio. Whole animal methane emissions, expressed as L/day or as a proportion of gross energy intake (%GEI), were unaffected by nitroethane treatment ( P > 0.05 ), and were not correlated ( P > 0.05 ) with ruminal methane-producing activity. These results demonstrate that oral nitroethane administration reduces ruminal methane-producing activity but suggest that a microbial adaptation, likely due to an in situ enrichment of ruminal nitroethane-reducing bacteria, may cause depletion of nitroethane, at least at the 1X administration dose, to concentrations too low to be effective. Further research is warranted to determine if the optimization of dosage of nitroethane or related nitrocompouds can maintain the enteropathogen control and anti-methanogen effect in fed steers.

A Risk Assessment Model for Campylobacter in Broiler Meat

A quantitative microbiological risk assessment model describes the transmission of Campylobacter through the broiler meat production chain and at home, from entering the processing plant until consumption of a chicken breast fillet meal. The exposure model is linked to a dose-response model to allow estimation of the incidence of human campylobacteriosis. The ultimate objective of the model is to serve as a tool to assess the effects of interventions to reduce campylobacteriosis in the Netherlands. The model describes some basic mechanistics of processing, including the nonlinear effects of cross-contamination between carcasses and their leaking feces. Model input is based on the output of an accompanying farm model and Dutch count data of Campylobacters on the birds' exterior and in the feces. When processing data are lacking, expert judgment is used for model parameter estimation. The model shows that to accurately assess of the effects of interventions, numbers of Campylobacter have to be explicitly incorporated in the model in addition to the prevalence of contamination. Also, as count data usually vary by several orders of magnitude, variability in numbers within and especially between flocks has to be accounted for. Flocks with high concentrations of Campylobacter in the feces that leak from the carcasses during industrial processing seem to have a dominant impact on the human incidence. The uncertainty in the final risk estimate is large, due to a large uncertainty at several stages of the chain. Among others, more quantitative count data at several stages of the production chain are needed to decrease this uncertainty. However, this uncertainty is smaller when relative risks of interventions are calculated with the model. Hence, the model can be effectively used by risk management in deciding on strategies to reduce human campylobacteriosis.

Effect of Oral Administration of Bismuth Compounds on Campylobacter Colonization in Broilers

Bismuth compounds have been used since the 18th century to treat gastrointestinal ailments in man. Colloidal bismuth subcitrate (De-Nol) is currently used in combination with antibiotics to reduce enteric Helicobacter pylori colonization as a treatment of stomach ulcers. We investigated whether bismuth citrate or its parent compound, colloidal bismuth subcitrate, would reduce colonization of the closely related foodborne pathogen, Campylobacter jejuni in chickens. In 2 studies, birds were either fed 0, 50, or 200 ppm bismuth citrate or bismuth subcitrate (De-Nol) for 10 or 21 d and were orally challenged with 7 combined strains of C. jejuni (n = 6 birds/treatment). For both treatment groups, cecal Campylobacter colonization was reduced when birds were fed 200 ppm for 10 d but not 21 d. For the 50 ppm treatment group, only birds dosed with bismuth citrate for 21 d demonstrated any reduction in cecal Campylobacter concentrations when compared with controls. These data suggest that bismuth citrate and colloidal bismuth subcitrate may reduce cecal colonization by Campylobacter in broilers, but these effects are inconsistent.

Bacteriocins Reduce Campylobacter Colonization and Alter Gut Morphology in Turkey Poults ,

Campylobacter is a leading cause of foodborne illness in the United States. Recent evidence has demonstrated that bacteriocins produced by Bacillus circulans and Paenibacillus polymyxa reduce cecal Campylobacter colonization in broiler chickens infected with Campylobacter jejuni. As Campylobacter coli is the most prevalent Campylobacter isolate recovered in turkeys, the objectives of the present study were to evaluate the efficacy of these bacteriocins against C. coli colonization and their influence on the gastrointestinal architecture of young turkeys. In 3 separate trials, a total of 135 day-of-hatch poults (n = 45/trial) were orally challenged on d 3 with approximately 106 cfu of a mixture of 3 C. coli isolates. Immediately before bacteriocin treatment (d 10), cecal Campylobacter concentrations averaged 1.1 × 107 cfu/ g of cecal contents (n = 15/trial). On d 10 to 12 posthatch, 2 bacteriocin treatment groups were given free access to feed supplemented with purified, microencapsulated bacteriocins, whereas the positive control treatment group had access to untreated feed (n = 10/treatment group per trial). At the end of the 3-d dosing period, ceca and duodenal loops were collected for analysis. In each of the 3 separate trials, treatment with bacteriocin eliminated detectable ceca Campylobacter concentrations (detection limit, 1 × 102 cfu/g of cecal contents) vs. controls (1.0 × 106 cfu of Campylobacter/g of cecal contents). Duodenum crypt depth and goblet cell numbers were also reduced in turkeys treated with either bacteriocin vs. controls (P < 0.05). The dynamic reduction in crypt depth and goblet cell density in turkeys dosed with bacteriocin may provide clues to how bacteriocins inhibit enteric Campylobacter.

Coliforms, Escherichia coli, Campylobacter, and Salmonella in a Counterflow Poultry Scalder with a Dip Tank

Suspended bacteria were enumerated in scald water and carcass rinse samples from a commercial broiler chicken processing plant with a multiple-tank, counterflow scalder. Coliforms, Escherichia coli, and Campylobacter were enumerated and the Most Probable Number (MPN) of salmonellae was determined in water samples from each of three scald tanks, from a dip tank located between defeathering machines, and in rinses of carcasses removed from the processing line immediately after defeathering. Mean coliform concentrations in Tanks 1, 2, and 3 were 4.6, 2.5, and 1.6 log (cfu/ml), respectively. E. coli 10 concentrations followed the same pattern with means of 4.4, 2.1, and 1.4 in Tanks 1, 2, and 3, respectively, with significant differences (P<.05) in the concentrations of both coliforms and E. coli between tanks. Mean Campylobacter concentration in four positive samples from Tank 1 was 4.0 log (cfu/ml), but only one water 10 sample from Tank 2 and none from Tank 3 were Campylobacter positive. Coliforms and E. coli were found in dip tank samples in only two instances, with no isolations of Campylobacter or salmonellae. Mean numbers of coliforms, E. coli, and Campylobacter in carcass rinses were 3.1, 2.7, and 3.3 log (cfu/ml). 10 Salmonellae were isolated from five of six water samples from Tank 1 with a mean MPN of 13.3/100mL, but were isolated from only three of six water samples from Tank 2 and two of six from Tank 3. Salmonellae were isolated from half (18/36) of all carcass rinses. Most bacteria suspended in scald water were found in the first tank, with no Campylobacter or salmonellae found in the dip tank. Counterflow, multiple-tank scalders appear to reduce the opportunity for cross-contamination during scalding.

Efficacy of Iron Chelators on Campylobacter Concentrations in Turkey Semen

Campylobacter is a leading bacterial cause of human foodborne infections in the United States. Recent studies suggest that the organism is highly prevalent in poultry semen and may contribute to vertical transmission between the breeder hen and offspring. Because Campylobacter requires iron for its growth and survival, the objective of this study was to determine if the addition of natural and synthetic chelators such as ovotransferrin, desferrioxaime, EDTA, or 2,2′-dipyridyl could reduce or eliminate Campylobacter in turkey semen. In a preliminary study without semen, a commercial poultry semen extender was supplemented with various concentrations of ovotransferrin, desferrioxaime, EDTA, or 2,2′-dipyridyl and inoculated with an average of 108 cfu/mL of a wild-type Campylobacter coli turkey semen isolate. At 6 and 24 h of storage at 4°C, a sample was taken from each treatment group and enumerated for Campylobacter. In all 3 trials, Campylobacter was undetectable (<102) in the commercial poultry semen extender supplemented with 20 mg/mL of 2,2′-dipyridyl. There were no differences observed in Campylobacter concentrations in the commercial poultry semen extender supplemented with ovotransferrin, desferrioxaime, or EDTA compared with unsupplemented controls. In a follow-up study, pooled semen samples were randomly collected from toms, diluted with a commercial poultry semen extender supplemented with 5, 10, or 20 mg/mL of 2,2′-dipyridyl and inoculated with an average of 108 cfu/mL of a wild-type C. coli turkey semen isolate. At 6 and 24 h of storage at 4°C, samples were taken from each treatment group, enumerated for Campylobacter, and evaluated for sperm viability. In all 3 trials, supplementing the commercial poultry semen extender with 20 mg/mL of 2,2′-dipyryidyl significantly reduced (3 to 4 logs) Campylobacter concentrations when compared with the positive controls. Sperm viability was also reduced with this treatment, and, therefore, the use of 2,2′-dipyridyl may not be a practical treatment for reducing Campylobacter in poultry semen.

Comparative, Collaborative, and On-Site Validation of a TaqMan PCR Method as a Tool for Certified Production of Fresh, Campylobacter -Free Chickens

Certified Campylobacter-free poultry products have been produced in Denmark since 2002, the first example of fresh (unprocessed and nonfrozen) chickens labeled "Campylobacter free." This success occurred partly through use of a 4-hour gel-based PCR testing scheme on fecal swabs. In this study, a faster, real-time PCR approach was validated in comparative and collaborative trials, based on recommendations from the Nordic system for validation of alternative microbiological methods (NordVal). The comparative real-time PCR trial was performed in comparison to two reference culture protocols on naturally contaminated samples (99 shoe covers, 101 cloacal swabs, 102 neck skins from abattoirs, and 100 retail neck skins). Culturing included enrichment in both Bolton and Preston broths followed by isolation on Preston agar and mCCDA. In one or both culture protocols, 169 samples were identified as positive. The comparative trial resulted in relative accuracy, sensitivity, and specificity of 98%, 95%, and 97%, respectively. The collaborative trial included nine laboratories testing neck skin, cloacal swab, and shoe cover samples, spiked with low, medium, and high concentrations of Campylobacter jejuni. Valid results were obtained from six of the participating laboratories. Accuracy for high levels was 100% for neck skin and cloacal swab samples. For low levels, accuracy was 100% and 92% for neck skin and cloacal swab samples, respectively; however, detection in shoe cover samples failed. A second collaborative trial, with an optimized DNA extraction procedure, gave 100% accuracy results for all three spiking levels. Finally, on-site validation at the abattoir on a flock basis was performed on 400 samples. Real-time PCR correctly identified 10 of 20 flocks as positive; thus, the method fulfilled the NordVal validation criteria and has since been implemented at a major abattoir.

El agente Campylobacter en la producción animal y las estrategias de control para reducir la incidencia de la campilobacteriosis humana

Campylobacteriosis is one of the most important bacterial food-borne illnesses in humans. One significant source of infection is the handling and consumption of poultry meat, although other sources also contribute considerably. Controlling Campylobacter in broilers reduces the human burden of illness. Broilers can easily become colonised with Campylobacter and preventive measures in primary production have a limited and unpredictable effect. Vaccination, competitive exclusion, bacteriophage therapy and the use of bacteriocins are not yet commercially available. However, measures in the slaughterhouse can reduce contamination in the final product. At present, the most promising control strategy is to keep colonised and non-colonised flocks separate during slaughter ('scheduled processing'). The virtually Campylobacter-free meat can supply the fresh poultry meat market, while the meat from infected flocks can be treated to reduce the Campylobacter concentration. Meat from infected flocks can be treated by freezing but chemical decontamination appears to be more cost effective. A variant of this scenario is to treat only highly contaminated meat. The authors conclude that, until new techniques become commercially available, scheduled processing is the most cost-effective approach. Finally, the authors describe trends in antimicrobial resistance in Campylobacter.

Combined use of microbiological and non-microbiological data to assess treatment efficacy

The treatment efficacy for reducing Campylobacter concentrations by a drinking water treatment plant was assessed using a stochastic Monte Carlo model. The goal of the study was to reduce uncertainty of the results by combining microbiological and non-microbiological data in an advanced treatment assessment. Combining raw water Campylobacter and E. coli data reduced the uncertainty on raw water (peak) concentrations five-fold. Similar improvement was achieved for rapid sand filtration. Ozone disinfection was modelled based on ozone concentrations, contact time and temperature. Since this data was available, whereas most microbiological analyses at this point were negative, uncertainty was reduced three-fold. The slow sand filtration assessment could not be improved; however, since previous steps contained less uncertainty, this did not increase uncertainty by much. The study showed that using appropriate data for each treatment step can greatly reduce uncertainty in treatment assessment.