Recovery of Campylobacter from feed using different enrichment media.

Studies were conducted to investigate the recovery of Campylobacter from feed. The impact of feed moisture, water activity, pH, number of background microflora and the use of different antibiotic supplements in Campylobacter enrichment broth (CEB) on Campylobacter recovery were evaluated in five studies. Broiler starter feed was inoculated with 104 -105 cfu of Campylobacter/g and stored at 24 °C and 43% RH. Enrichment culture was conducted on the day of inoculation or 24 h post inoculation and every 48 h of storage thereafter for 14 d. Feed moisture, water activity, pH and level of background microflora were not correlated with Campylobacter recovery. The incubation of feed in CEB with no antibiotic supplement resulted in the number of background microflora increasing to 109 cfu/g and the pH of the media decreasing to pH 4-5 impacting recovery. Addition of certain antimicrobial supplements to CEB reduced background microflora growth and maintained a near neutral pH. Campylobacter was recovered up to 10 days post inoculation when using CEB containing antibiotic supplements compared to 1 day in CEB. These findings suggest that Campylobacter can be recovered from feed and the type of antimicrobial supplement utilized influences recovery by controlling extraneous microbial growth which occurs during enrichment.

Recovery of thermophilic Campylobacter by three sampling methods from river sites in Northeast Georgia, USA, and their antimicrobial resistance genes.

Sixteen sites in the watershed of the South Fork of the Broad River (SFBR) in Northeastern Georgia, USA, were sampled in two seasons to detect Campylobacter. Sites were classified as mostly influenced by forest, pasture, wastewater pollution control plants (WPC) or mixed use. Sampling was repeated in the late spring and late fall for 2 years for a total of 126 samples. Free-catch water and sediment grab samples were taken at each site; Moore's swabs were placed for up to 3 days at most sites. A total of 56 isolates of thermophilic Campylobacter were recovered. Thirteen samplings were positive by two or three methods, and 26 samplings were positive by only one method; once by Moore's swab only and 25 times by free-catch water only. Campylobacter was detected at 58% of cattle pasture sites, 30% of forested sites and 81% of WPC sites. Twenty-one of the isolates carried antimicrobial resistance genes, mostly blaOXA-61. Free-catch water samples were more efficient than Moore's swabs or sediment samples for recovery of Campylobacter, which was more likely to be detected in streams near cattle pastures and human communities than in forested land. SIGNIFICANCE AND IMPACT OF THE STUDY: The role of environmental water in transmitting Campylobacter was investigated, and methods for recovery of the organism were compared. The sequence types of recovered Campylobacter correlated with adjacent land use without regard to the method used to isolate the organisms. Sequence types and antimicrobial resistance genes associated with cattle were most prevalent near pastures. Even though types were recurrent at a given site, types appeared to be lost or replaced as the water flowed downstream.

Optimizing buffering chemistry to maintain near neutral pH of broiler feed during pre-enrichment for Salmonella.

Salmonella is a human pathogen that can accompany live broilers to the slaughter plant, contaminating fully processed carcasses. Feed is one potential source of Salmonella to growing broilers. Monitoring feed for the presence of Salmonella is part of good agricultural practice. The first step in culturing feed for Salmonella (which may be at low numbers and sub-lethally stressed) is to add it to a pre-enrichment broth which is incubated for 24 h. During the course of pre-enrichment, extraneous bacteria metabolize carbohydrates in some feed and excrete acidic byproducts, causing the pH to drop dramatically. An acidic pre-enrichment pH can injure or kill Salmonella resulting in a failure to detect, even if it is present and available to infect chickens. The objective of this study was to test an array of buffering chemistries to prevent formation of an injurious acidic environment during pre-enrichment of feed in peptone water. Five grams of feed were added to 45 mL of peptone water buffered with carbonate, Tris pH 8, and phosphate buffering ingredients individually and in combination. Feed was subjected to a pre-enrichment at 35°C for 24 h; pH was measured at 0, 18, and 24 h. Standard phosphate buffering ingredients at concentrations up to 4 times the normal formulation were unable to fully prevent acidic conditions. Likewise, carbonate and Tris pH 8 were not fully effective. The combination of phosphate, carbonate, and Tris pH 8 was the most effective buffer tested. It is recommended that a highly buffered pre-enrichment broth be used to examine feed for the presence of Salmonella.

Campylobacter spp. recovered from the Upper Oconee River Watershed, Georgia in a 4-year study.

Waterways should be considered in the migration routes of Campylobacter, and the genus has been isolated from several water sources. Inferences on migration routes can be made from tracking genetic types in populations found in specific habitats and testing how they are linked to other types. Water samples were taken over a 4-year period from waterways in the Upper Oconee River Watershed, Georgia, to recover isolates of thermophilic Campylobacter. The isolates were typed by multilocus sequence typing (MLST) and analyzed to determine the overall diversity of Campylobacter in that environment. Forty-seven independent isolates were recovered from 560 samples (8.4 %). Two (~4 %) isolates were Campylobacter coli, three (~6 %) isolates were putatively identified as Campylobacter lari, and the remaining 42 (~90 %) were Campylobacter jejuni. The C. jejuni and C. coli isolates were typed by the Oxford MLST scheme. Thirty sequence types (STs) were identified including 13 STs that were not found before in the MLST database, including 24 novel alleles. Of the 17 previously described STs, 10 have been isolated from humans, 6 from environmental water, and 6 from wild birds (five types from multiple sources). Seven sites had multiple positive samples, and on two occasions, the same ST was isolated at the same site. The most common type was STST61 with four isolates, and the most common clonal complex was CC179 with nine isolates. CC179 has been commonly associated with environmental water. Although some Campylobacter STs that were found in the Oconee River engage in widespread migration, most are tightly associated with or unique to environmental water sources.

Enrichment and Direct Plating for Detection of Campylobacter in Chicken Liver Rinse and Exudate.

ABSTRACT: Foodborne campylobacteriosis has been traced to undercooked chicken liver dishes; thus, it is important to use the best available culture methods when testing for the presence of Campylobacter. We compared two Campylobacter enrichment broths-Bolton formulation and Neogen formulation-in combination with three selective plating media-Campy-Cefex, Campy-Line and RF Campylobacter agars-for detection of Campylobacter from fresh retail chicken livers. In each of three experiments, nine replicate tubs of chicken livers were sampled by drawing exudate and a pooled rinse of five whole liver lobes. Results are reported as number positive and compared by Fisher's exact test. In experiment 1, no combination of enrichment and plating media significantly outperformed another for detection of Campylobacter (P > 0.05); all tubs were found to include Campylobacter in both exudate and liver rinse. In experiment 2, serial dilutions of samples were plated before and after enrichment. Exudate was found to be significantly more likely than rinse to support detection of Campylobacter by direct plating (P < 0.05); most exudate samples included at least 10 CFU Campylobacter per mL. Enrichment improved detection from rinse, but not exudate; all enrichment and plating combinations resulted ≥1,000 CFU/mL from most enriched samples. In experiment 3, samples were diluted before enrichment to determine effect of enrichment on ever lower numbers of Campylobacter. Enrichment did not improve recovery of Campylobacter from exudate or undiluted rinse (P > 0.05). However, when rinse samples were diluted to lower Campylobacter numbers, enrichment improved detection (P < 0.05). Overall, all media combinations tested were equivalent for detection of Campylobacter from chicken livers; sensitivity for detection seemed to be increased by using liver exudate compared with a pooled rinse of liver lobes.

Campylobacter, Salmonella, and Escherichia coli on broiler carcasses subjected to a high pH scald and low pH postpick chlorine dip.

The objective of this study was to determine the individual and combined effects of a high pH scald and a postpick chlorine dip on bacteria present on broiler carcasses. In each of 3 replications, a flock was sampled at several sites within a commercial broiler processing plant. Carcasses were sampled by whole carcass rinse before and after treated scalding at mean pH 9.89 or control scalding at mean pH 6.88. Other carcasses from the same flock run on both the treated and control scald lines were collected and sampled before and after a chlorine dip tank operated at mean total chlorine level of 83.3 mg/kg and pH 6.04. Rinses were cultured for numbers of Campylobacter and Escherichia coli and presence or absence of Salmonella. High pH scald was more effective than standard scald to lessen the prevalence and numbers of Campylobacter on broiler carcasses; a lower prevalence was maintained through the postpick chlorine dip tank. The pH of the scald tank made no difference in numbers of E. coli recovered from broiler carcasses at any tested point on the processing line. High pH scald was not more effective than standard scald to lessen Salmonella prevalence. Furthermore, it is unclear why the postpick chlorine dip effectively lessened Salmonella prevalence on only the control scald line. Although no evidence exists that these treatments have an additive effect when used in series, each treatment shows some promise individually. Further optimization may result in more effective decontamination of broiler carcasses.

Presence of Bacterial Pathogens and Levels of Indicator Bacteria Associated with Duck Carcasses in a Commercial Processing Facility.

ABSTRACT: Little information has been published on the microbiological aspects of U.S. commercial duck processing. The objective of this study was to measure prevalence and/or levels of bacteria in duck samples representing the live bird and partially or fully processed oven-ready duck meat. At 12 monthly sampling times, samples were collected at six sites along the processing line in a commercial duck slaughter plant. Crop and cecum samples were collected at the point of evisceration. Whole carcass rinse samples were collected before and after carcass immersion chilling plus application of an antimicrobial spray. Leg quarters were collected from the cut-up line before and after application of an antimicrobial dip treatment. All samples (five from each site per monthly replication) were directly plated and/or enriched for Salmonella and Campylobacter. For the last 10 replications, carcass and leg quarter rinse samples were also evaluated for enumeration of total aerobic bacteria, Escherichia coli, and coliforms. Most cecum, crop, and prechill carcass rinse samples were positive for Campylobacter (80, 72, and 67%, respectively). Carcass chilling and chlorinated spray significantly lowered Campylobacter prevalence (P < 0.01), and even fewer leg quarters were positive for Campylobacter (P < 0.01). Passage through a chlorinated dip did not further reduce Campylobacter prevalence on leg quarters. Salmonella was infrequently found in any of the samples examined (≤10%). Total aerobic bacteria, coliforms, and E. coli levels were reduced (P < 0.01) on whole carcasses by chilling but were not different after cut-up or leg quarter dip treatment. Overall, current commercial duck processing techniques as applied in the tested plant were effective for reducing the prevalence and levels of Campylobacter on duck meat products.

Research Note: Evaluation of several inoculation procedures for colonization of day-old broiler chicks with Salmonella Heidelberg.

Before starting a study with many birds, it helps to know the method of chick inoculation. The objective was to compare 3 methods of Salmonella challenge (oral gavage [OR], intracloacal inoculation [IC], and seeder bird [SB]). Day-old broiler chicks (n = 100) were inoculated with 106 colony forming units (CFU) per chick of a marker strain of Salmonella Heidelberg (SH) with each route of inoculation. Chicks (n = 25) inoculated by each route were placed in floor pens on fresh pine shavings litter. For the seeder batch, 5 colonized chicks, each orally gavaged with 106 CFUs, were placed with 20 pen mates. Two weeks after inoculation, 10 birds from each pen and the 5 inoculated seeder birds were euthanized, the ceca were aseptically removed and macerated with a rubber mallet and weighed, and 3 times (w/v) buffered peptone was added and stomached for 60 s. Serial dilutions were made and plated onto Brilliant Green Sulfa plates containing 200 ppm nalidixic acid. Plates were incubated along with the stomached ceca for 24 h at 37°C. If no colonies appeared on the plates, an additional plate was streaked from the preenriched bag and incubated for 24 h at 37°C. In addition to all seeder birds being positive, the number of SH-positive birds out of 20 sampled in each group was 13, 17, and 7 for OR, IC, and SB, respectively. The level of SH per g of ceca and cecal contents was log (SE) 3.0 (0.7), 2.0 (0.4), and 2.6 (0.4) for OR, IC, and SB, respectively. After enrichment, the number of colonized birds out of 20 was 18, 20, and 10 for OR, IC, and SB, respectively. In conclusion, this study suggests that IC is the method to use to ensure most of the challenged birds are colonized. However, if you prefer to have a smaller percentage of the birds colonized with higher levels, then OR might be better.

Genetic diversity of Campylobacter on broiler carcasses collected preevisceration and postchill in 17 U.S. poultry processing plants.

Campylobacter jejuni and Campylobacter coli are the most important human enteropathogens among the campylobacters. The objective of this study was to determine how diversity in Campylobacter populations found on chicken carcasses collected from 17 broiler processing plants in the United States is impacted by processing. Genetic diversity was determined for up to four isolates per carcass by sequencing the short variable region (SVR) of the flaA locus. On 70% of Campylobacter-positive carcasses, all isolates were indistinguishable by flaA SVR typing. The genetic diversity of Campylobacter decreased as carcasses proceeded through processing; Campylobacter populations obtained early in processing where carcasses are moved from the kill line to the evisceration line (rehang) were significantly more genetically diverse (P < 0.05) than those from carcasses sampled postchill (diversity indices of 0.9472 and 0.9235, respectively). Certain Campylobacter subtypes were found only at rehang and not at postchill. Other subtypes were found at postchill and not at rehang. These data suggest that some subtypes may not be able to survive processing, whereas others may persist on the carcass or within the equipment despite stressors encountered in the processing environment.

Campylobacter coli naturally resistant to elevated levels of gentamicin as a marker strain in poultry research.

Campylobacter inoculation studies are limited without a suitable marker strain. The lurpose of this study was to screen Campylobacter strains (n=2073) obtained from poultry carcass rinses through the Centers for Disease Control and Prevention's National Antimicrobial Resistant Monitoring System for resistance to gentamicin and evaluate one strain's efficacy as a marker. A C. coli strain was found resistant to gentamicin at >32 microg/ml. Gentamicin was incorporated into media (Campy-Cefex agar, Brucella agar, and blood agar) from 0 to 1000 microg/ml, and the upper level of gentamicin resistance was determined. C. coli strain's upper level of growth on Campy-Cefex plates, blood agar plates, and Brucella agar plates was 400, 300, and 200 pg/ml, respectively. Ceca and postpick carcass rinses were obtained and streaked onto Campy-Cefex agar at the above gentamicin levels to evaluate background microflora exclusion. Campy-Cefex agar containing gentamicin at 100 ag/ml prevented from the ceca, and reduced from the rinse, background microflora. The C. coli strain was orally or intracloacally inoculated into chicks. At 1, 3, and 6 weeks of age, inoculated broilers were removed and several tissue types sampled for the presence of the marker strain. At 6 weeks of age, 10 additional noninoculated penmates were sampled. The C. coli strain colonized chicks, disseminated to body tissues, colonized penmates, and persisted throughout the 6-week grow-out. The C. coli strain's unique characteristic, being resistant to high levels of gentamicin, allows for a marker that can be used in a wide range of Campylobacter research projects.

Evaluation of TECRA broth, Bolton broth, and direct plating for recovery of Campylobacter spp, from broiler carcass rinsates from commercial processing plants.

The purpose of this study was to compare a conventional culture broth method (Bolton enrichment), a newly developed proprietary broth method (TECRA Campylobacter enrichment), and direct plating for recovery of Campylobacter spp. from chicken carcass rinsates. Whole carcass rinses were taken from 140 carcasses at rehang (immediately after defeathering but before evisceration) and from 140 carcasses at postchill from eight different processing plants in the United States. The rinsate samples were packed in ice and shipped overnight to the laboratory. Aliquots of the rinsate were transferred into Bolton and TECRA enrichment broths and were direct plated. Standard laboratory procedures with Campy-cefex plates were followed for recovery of Campylobacter spp. For rehang carcasses, 94% were positive for Campylobacter spp. with the TECRA enrichment broth and 74% were positive with the Bolton enrichment broth. For postchill carcasses, 74% were positive for Campylobacter spp. with the TECRA enrichment broth and 71% were positive with the Bolton enrichment broth. Compared with the Bolton enrichment broth, TECRA enrichment broth significantly suppressed non-Campylobacter microflora (P < 0.05). Overall, TECRA enrichment broth yielded an 11% higher total number of Campylobacter-positive samples compared with the Bolton enrichment broth. Campylobacter spp. detection in postchill samples was significantly greater (P < 0.05) by enrichment (84%) than by direct plating (19%). The high number of Campylobacter-positive samples obtained with all procedures indicated that 99% of the carcass rinsates obtained at rehang and 84% obtained at postchill contained Campylobacter spp.

Prevalence, serotype, and antimicrobial resistance of Salmonella on broiler carcasses postpick and postchill in 20 U.S. processing plants.

The objective of this study was to measure the effect of broiler processing on the prevalence, serotype, and antimicrobial resistance profiles of salmonellae. Twenty U.S. commercial processing plants representing eight integrators in 13 states were included in the survey. In each of four replications, 10 carcasses from one flock were collected at rehang and 10 more carcasses were collected at postchill; each carcass was sampled by whole-carcass rinse. Salmonella organisms were isolated from carcass rinses by standard cultural techniques, serotypes were determined, and the resistance to 15 antimicrobials was measured. Overall, Salmonella was detected on 72% of carcasses at rehang (ranging from 35 to 97%) and on 20% of carcasses postchill (ranging from 2.5 to 60%). In every instance, a significant (P < 0.05) decrease in Salmonella prevalence was noted between rehang and postchill. The four most common serotypes, accounting for 64% of all Salmonella isolates, were Kentucky, Heidelberg, Typhimurium, and Typhimurium var. 5-; most isolates of Kentucky (52%), Heidelberg (79%), and Typhimurium (54%) serotypes were susceptible to all antimicrobial drugs tested. However, only 15% of the Typhimurium var. 5- isolates were pansusceptible; more than one-half of the isolates of this serotype were resistant to three or more drugs. No isolate of any serotype exhibited resistance to amikacin, ceftriaxone, ciprofloxacin, or trimethoprim-sulfamethoxazole. These data demonstrate that although processing lessens carcass contamination with Salmonella, antimicrobial-resistant isolates may still be present.

Effect of dry-air chilling on sensory descriptive profiles of cooked broiler breast meat deboned four hours after the initiation of chilling.

The objective of this study was to evaluate the effect of a dry air-chilling (AC) method on sensory texture and flavor descriptive profiles of broiler pectoralis major (fillet) and pectoralis minor (tender). The profiles of the muscles immersion-chilled and deboned at the same postmortem time and the profiles of the muscles hot-boned (or no chill) were used for the comparison. A total of 108 eviscerated carcasses (6-wk-old broilers) were obtained from a commercial processing line before the chillers. Carcasses were transported to a laboratory facility where they were either i) chilled by a dry AC method (0.7 degrees C, 150 min in a cold room), ii) chilled by immersion chilling (IC; 0.3 degrees C, 50 min in a chiller), or iii) not chilled (9 birds per treatment per replication). Both IC and AC fillets and tenders were removed from the bone at 4 h after the initiation of chilling (approximately 4.75 h postmortem) in a processing area (18 degrees C). The no-chill muscles were removed immediately upon arrival. The sensory properties (21 attributes) of cooked broiler breast meat were evaluated by trained panelists using 0- to 15-point universal intensity scales. The average intensity scores of the 9 flavor attributes analyzed ranged from 0.9 to 4.0. Regardless of breast muscle type, there were no significant differences in sensory flavor descriptive profiles between the 3 treatments. The average intensity scores of the 12 texture attributes ranged from 1.5 to 7.5 and there were no significant differences between the AC and IC samples. The average intensity scores of the texture attributes, cohesiveness, hardness, cohesiveness of mass, rate of breakdown, and chewiness of the no chill fillets and tenders were significantly higher than those of either of the chilled samples. These results demonstrate that chicken breast meat from AC retains sensory flavor profile characteristics but AC results in sensory texture profile differences when compared with no-chill meat. Sensory flavor and texture profiles of AC broiler breast meat do not differ from those of IC samples when the muscles are deboned at the same time after the initiation of chilling.

Campylobacter subtypes detected in broiler ceca and livers collected at slaughter1.

Foodborne campylobacteriosis has been linked to undercooked chicken liver. We have detected Campylobacter in chicken livers available at retail. The objective of the current project was to determine the prevalence and subtype of Campylobacter associated with livers and ceca of the same broiler carcasses at commercial slaughter. Within 2 min of commercial evisceration, we collected liver and ceca of one broiler carcass from each of 70 discreet flocks over a 12-mo period. Liver surface, liver internal tissue, and cecal contents were cultured for Campylobacter using standard methods. One example of the predominant colony type was selected from each positive sample for whole genome sequencing and multilocus sequence typing. We detected Campylobacter in at least one sample from 58 of 70 (83%) carcasses/flocks; 41 ceca, 57 liver surface samples, and 19 liver internal tissue samples were positive. For 11 of 18 carcasses from which all samples were positive, the predominant colony types were indistinguishable. However, some carcasses did have multiple subtypes of Campylobacter. Of carcasses with Campylobacter on the surface of the liver and within the ceca, it was more likely that the subtypes be the same than different (P < 0.01). However, Campylobacter subtypes detected in internal liver tissue were not more likely to be the same as those detected in ceca (P > 0.05). We detected different subtypes of Campylobacter from internal liver tissue and liver surface of seven broiler carcasses/flocks. Livers from a large percentage of broiler carcasses/flocks can have one or more subtypes of Campylobacter.

Effect of chemical sanitizers with and without ultrasonication on Listeria monocytogenes as a biofilm within polyvinyl chloride drain pipes.

As part of a biofilm in a floor drain, Listeria monocytogenes is exceedingly difficult to eradicate with standard sanitizing protocols. The objective of these studies was to test the use of ultrasonication to break up biofilm architecture and allow chemical sanitizers to contact cells directly. L. monocytogenes biofilms were created in model polyvinyl chloride drain pipes. Chemical sanitizers (quaternary ammonium, peroxide, or chlorine) were applied to the drain pipes with and without a 30-s ultrasonication treatment. Controls using sterile water were included for comparison. L. monocytogenes cells were enumerated from the liquid in the drain and the inside wall surface of the pipe. All chemicals lowered numbers of planktonic cells from 6.6 log CFU/ml in the water control to < 100 CFU/ml. Attached cells were also affected by the chemical sanitizers. Approximately 6.0 log CFU/cm2 of the inner wall surface was detected in water control pipes, and ultrasonication did not lower these numbers. With or without ultrasonication, the peroxide-based sanitizer was effective for reducing the numbers of attached L. monocytogenes cells, resulting in approximately 2.0 log CFU/cm2. Both the chlorine- and quaternary ammonium-based sanitizers reduced the number of attached L. monocytogenes cells to a lesser degree, resulting in 4.2 to 4.4 log CFU/cm2. However, addition of ultrasonication improved the performance of both these sanitizers, causing a further reduction to 3.1 and 2.9 CFU/ cm2 for quaternary ammonium- and chlorine-based chemicals, respectively. These results indicate that a peroxide-based sanitizer alone can be very effective against biofilm L. monocytogenes in drain pipes, and the addition of ultrasonication can improve the effectiveness of chlorine or quaternary ammonium sanitizers.

The effect of chilling in cold air or ice water on the microbiological quality of broiler carcasses and the population of Campylobacter.

Cold air or ice water can be used to chill poultry carcasses after slaughter. The objective of this study was to compare the effect of 2 chill methods on broiler carcass bacteria. Broiler carcasses were cut in half along the dorsal-ventral midline; one half was subjected to an ice-water immersion chill in an agitated bath for 50 min, whereas the reciprocal half was subjected to an air chill in a 1 degrees C cold room for 150 min. Total aerobic bacteria, coliforms, Escherichia coli, and Campylobacter were enumerated from half-carcass rinses. Species of Campylobacter isolates was determined by a commercial PCR method, which was followed by molecular subtyping with pulsed-field gel electrophoresis and determination of antimicrobial susceptibility to 9 drugs. Although significantly fewer of each bacterial type were detected per milliliter from immersion-chilled carcasses than from air-chilled carcasses, in each case the difference was less than 1 log(10) cfu/mL. Chilling method did not affect species; both Campylobacter jejuni and Campylobacter coli were recovered. Results of pulsed-field gel electrophoresis subtyping did not suggest that either chilling method selected for any specific subtypes; most subtypes were found on carcass halves used for both the air chill and water immersion chill. Resistance to 2 antimicrobial drugs was noted in 9 C. coli isolates, 6 from air-chilled carcass halves and 3 from immersion-chilled carcass halves. These data showed that immersion-chilled carcasses had lower numbers of bacteria; however, the difference was not large and may have been due to simple dilution. Both methods were effective for lowering carcass temperature, and neither chilling method seemed to select for specific species, subtypes, or antimicrobial-resistant Campylobacter.

Multilocus Sequence Subtypes of Campylobacter Detected on the Surface and from Internal Tissues of Retail Chicken Livers.

Foodborne campylobacteriosis has been traced to undercooked chicken liver. The objectives of this study were to determine the prevalence of Campylobacter associated with chicken livers at retail and to determine which subtypes are detected on the surface and in the internal tissues of the livers. Fifteen packages of fresh chicken livers, each representing a unique combination of processing plant and sell-by date, were collected at each of three retail grocery stores in Georgia. Three intact, undamaged livers per container ( n = 45) were selected and sampled using each three methods: outside swab, inside swab accessed by pressing through a heat-sterilized outer surface, and whole liver blended in enrichment broth. Each liver sample with 0.1 mL of exudate from packages was cultured for Campylobacter by plating on Campy-Cefex agar. The most prevalent Campylobacter colony type from each positive sample was subjected to whole genome sequencing and multilocus sequence typing. Campylobacter was detected in at least one sample from every package. Surface swabs were positive for 29 of 45 livers, but significantly fewer swabs of internal tissue were positive, 14 of 45 ( P < 0.01). Campylobacter was detected in 30 of 45 blended whole liver samples. Multiple subtypes were detected from eight livers. In four livers, a different subtype was dominant on the surface than was dominant internally. In one liver, three subtypes were detected. Various subtypes of Campylobacter can be readily isolated from fresh retail chicken livers; therefore, undercooked chicken livers pose a food safety risk.

Prevalence and numbers of Campylobacter on broiler carcasses collected at rehang and postchill in 20 U.S. processing plants.

Campylobacter is a human pathogen associated with chicken and chicken meat products. This study was designed to examine the prevalence and number of Campylobacter on broiler chicken carcasses in commercial processing plants in the United States. Carcass samples were collected from each of 20 U.S. plants four times, roughly approximating the four seasons of 2005. At each plant on each sample day, 10 carcasses were collected at rehang (prior to evisceration), and 10 carcasses from the same flock were collected postchill. A total of 800 carcasses were collected at rehang and another 800 were collected postchill. All carcasses were subjected to a whole-carcass rinse, and the rinse diluent was cultured for Campylobacter. The overall mean number of Campylobacter detected on carcasses at rehang was 2.66 log CFU per ml of carcass rinse. In each plant, the Campylobacter numbers were significantly reduced by broiler processing; the mean concentration after chill was 0.43 log CFU/ml. Overall prevalence was also reduced by processing from a mean of > or =30 of 40 carcasses at rehang to > or =14 of 40 carcasses at postchill. Seven different on-line reprocessing techniques were applied in the test plants, and all techniques resulted in <1 log CFU/ml after chilling. Use of a chlorinated carcass wash before evisceration did not affect the postchill Campylobacter numbers. However, use of chlorine in the chill tank was related to lower numbers on postchill carcasses. Overall, U.S. commercial poultry slaughter operations are successful in significantly lowering the prevalence and number of Campylobacter on broiler carcasses during processing.

Germicidal ultraviolet light to lower numbers of Listeria monocytogenes on broiler breast fillets.

Raw broiler breast fillets were subjected to germicidal ultraviolet (UV) light (dose of 1,000 microW/cm(2) for 5 min at a wavelength of 254 nm) to evaluate its potential to reduce Listeria monocytogenes numbers on raw product before shipment to a poultry further-processing plant. Boneless, skinless breast fillets were inoculated with 4 different strains of L. monocytogenes 5 min before treatment. After the UV treatment, breast fillets were stored at 4 degrees C for 24 h. Enumeration of remaining L. monocytogenes was performed using the spread plate method on modified Oxford agar. An approximate 2-log reduction in viable L. monocytogenes was observed with all 4 strains on UV-treated breast fillets as compared with the nontreated breast fillets. The UV treatment caused only slight changes in meat color (lightness, redness, and yellowness) on day of treatment or after 7 d of storage. This study suggests that UV treatment of raw breast fillets at a slaughter plant can significantly reduce L. monocytogenes without negatively affecting meat color. This process could be used to reduce the negative effect of raw poultry as a transmission vector of L. monocytogenes into a poultry further-processing plant.

Subtherapeutic tylosin phosphate in broiler feed affects Campylobacter on carcasses during processing.

Tylosin phosphate is an antimicrobial drug approved for use in broiler feed at subtherapeutic levels for growth promotion. Erythromycin is often the drug of choice for treating humans with campylobacteriosis. Both tylosin and erythromycin are classified as macrolide drugs and cross-resistance between these antimicrobials occurs. Commercial broiler chicks were placed in isolation grow-out chambers and colonized with Campylobacter jejuni. From 14 d of age through grow-out, broilers were fed ad libitim a diet that included 22 ppm of tylosin phosphate (20 g/ton). Control broilers received the same diet without tylosin phosphate. At 42 d of age, broilers were processed in a pilot plant with equipment that closely modeled commercial conditions. Carcass rinses were collected after feather removal, after inside and outside washing, and after immersion chilling. Campylobacter numbers recovered from carcasses after feather removal did not differ according to feed type (3.53 log cfu/mL of rinse for control carcasses, and 3.60 log cfu/mL of rinse for those fed medicated feed). Likewise, medicated feed did not affect Campylobacter numbers on carcasses after inside-outside washing (3.11 and 3.07 log cfu/mL of rinse). However, carcasses of broilers fed tylosin phosphate had lower numbers of Campylobacter after chilling (1.45 log cfu/mL of rinse) than control carcasses (2.31 log cfu/mL of rinse). No Campylobacter isolated from control carcasses were resistant to erythromycin; all Campylobacter recovered from carcasses fed tylosin phosphate were resistant to erythromycin. Application of tylosin phosphate in feed results in lower numbers of Campylobacter on chilled carcasses; however, the Campylobacter that do remain are resistant to erythromycin.