Advanced Food and Nutritional Sciences

Short Key Title: Adv Food Nutri Sci
Start Year: 2016

Landrum MA, NA Cox, JL Wilson, DE Cosby, ME Berrang, GR Gamble, MJ Da Costa, GM Pesti and A Hinton, Jr.
. 950 College Station Road Athens, GA 30605. T.: (706)546-3484 F.: (706)546-3633
Keywords: Campylobacter, CMS PoultrypHresh™, broiler, carcass rinse, immersion.

Campylobacter is a serious foodborne pathogen for which the USDA-FSIS has established stricter performance standards. Processors must establish effective treatment plans to lower the microbial levels of Campylobacter to meet these regulations. This study evaluated the low pH processing aid - CMS PoultrypHresh™ to reduce Campylobacter on carcasses (3 groups of 6) collected prior to the chiller that were individually placed into a 38 L container with either 20 L tap water (pH = 7.3) or 20 L of CMS PoultrypHresh™ solution (pH = 1.4) with air agitation. An untreated group was the control. After treatment, drained carcasses were placed into a plastic bag, and rinsed by hand in 400 mL of buffered peptone for 60 s. Rinsates were cultured for Campylobacter by direct plating on Campy-cefex agar and enrichment in Bolton's broth. If no Campylobacter was detected by direct plating, the incubated broth was plated and incubated under the same conditions. Confirmed Campylobacter were detected on 30/36 (83.3%) untreated carcasses, on 25/36 (69.4%) water treated carcasses, and on 2/36 (5.6%) of CMS PoultrypHresh™ treated carcasses. This treatment may be an option for processors to meet requirements and minimize Campylobacter to avoid regulatory action from FSIS.

Article Information

Identifiers and Pagination:
First Page:7
Last Page:13
Publisher Id:Adv Food Nutri Sci (2018 ). 3. 7-13
Article History:
Received:April 23, 2018
Accepted:June 8, 2018
Collection year:2018
First Published:June 11, 2018


USDA-Food Safety and Inspection Service (FSIS) has established new performance standards for the presence of Campylobacter and Salmonella on fresh poultry. Processors have responded by continuing to develop and test intervention methods that lower the prevalence of foodborne microorganisms in order to be in compliance with new regulations. Campylobacter is a major pathogen of concern which causes foodborne illness with gastrointestinal symptoms to more than 1.3 million people in the U.S. each year. [1] Campylobacter is most often reported being associated with poultry and has been found on as many as 88% of chicken carcasses. [2] Through the past seven years, USDA-FSIS has encouraged greater control of food safety measures, improved record keeping, proper labeling requirements, correct testing methods, and higher performance standards. [3] These regulations were made to reduce illnesses associated with foodborne pathogens found in poultry products and prevent an estimated 50,000 illnesses each year. [3] More specifically, recent regulatory changes aim to achieve a 32% reduction of illnesses from Campylobacter alone. [3] It is essential for broiler processing companies to establish an effective treatment plan to lower microbial levels of Campylobacter to pass rigorous performance standards set by USDA-FSIS. FSIS predicts that 46% of broiler companies will not be able to meet proposed 2016 performance standards for Campylobacter. [4]

Under regulations enacted in 2016, processing facilities are designated into categories 1, 2, 3, 4 or 5 by whether their microbial performance is consistent, variable, highly variable, passing, or failing to achieve FSIS Campylobacter standards. [5] Sampling to evaluate microbial control methods will become routine throughout the year, rather than infrequent sampling on consecutive days. [3] Results will be reported to the public, heightening the importance of reducing microbial levels to reach set standards and avoiding consumer rejection. Being listed on the FSIS website as “failing” to meet standards may have consequences for a company’s brand loyalty. The objective of this study was to evaluate a twenty second air agitated immersion in a proprietary antimicrobial treatment, PoultrypHresh™, at a pH of 1.4 as a means to reduce or eliminate viable Campylobacter on whole broiler carcasses.


Materials and Methods

Treatment Procedures

Eighteen whole, pre-chill broiler carcasses were randomly collected from the evisceration line of a commercial processing facility prior to chilling, individually bagged, placed on ice and transported to the laboratory. Within 60 min, six whole carcasses were individually dipped for 20 s into 38 L, high density polyethylene plastic containers holding 20 L of tap water at a pH of approximately 7.3. An additional 6 whole carcasses were dipped into similar containers for 20 s with 20 L of PoultrypHresh™ solution at a pH of 1.4. Air agitation of the solution in each bucket was achieved by pumping air (50 psi) from a compressor, through a six way manifold to the bottom of each container by means of plastic tubing (1/4” ID, McMaster-Carr, Elmherst, IL) which was secured to the side of the bucket by a metal pipe placed such that the tubing air exit was at the bottom of each container. After treatment (20 s immersion with agitation), each carcass was removed, and allowed to drip for 5 s before being subjected to a whole carcass rinse procedure in 400 mL of buffered peptone water (BPW) for 60 s using a mechanical rinsing machine. [6] The final six carcasses served as untreated controls, receiving no dip treatment before whole carcass rinse sampling, as described above. Six replications were conducted on 6 separate days, n=36 per treatment.

Campylobacter Detection Methods

Rinsates from each whole carcass were treated as described in the FSIS microbiology laboratory guidebook. [7] Initial rinses were used to spread 0.25 mL on four Campy-Cefex plates which were incubated microaerobically (5% O2, 10% CO2, 85% N2) for 48 h at 42°C. Thirty mL of each carcass rinsate was also placed into a tissue culture flask with 30 mL of 2X Bolton’s broth (2X-BEB) and incubated concurrently with the direct plates under the same conditions. Incubated enrichment broth was plated onto a Cefex agar plate and incubated for 48 h under the same conditions.  After incubation, plates were examined for typical Campylobacter colonies which were confirmed as members of the genus Campylobacter by observation of typical cellular morphology, motility under phase contrast microscopy and by use of a latex agglutination test kit (Microgen Bioproducts Ltd, Camberley, U.K.).  

Statistical Analysis

The study was replicated six times using naturally contaminated Campylobacter carcasses, totaling 36 whole birds for each treatment (N=108, n=36). In each replicate, 6 untreated, 6 tap waters treated, and 6 treated with PoultrypHresh™ at pH 1.4 were used. Each sample was recorded as either positive or negative for the presence of Campylobacter and data was analyzed using the Chi Square test for independence. Data was analyzed using a generalized mixed model (binomial distribution) where treatment was considered as a fixed effect and the treatment nested within the days of collection as a random effect. The GLIMMIX Procedure of SAS (SAS Inst. Inc., Cary, NC) was used: Proc GLIMMIX Data=campy; Class day rep trt; Model status = trt /dist=bin link=logit solution; Random day(trt).

Results and discussion

There was no difference in the presence of Campylobacter on the control or water treated samples from replication to replication. The covariance of treatment within replications was not significant (p=0.427). Overall, data from all six replicates indicated that 30/36 (83.3%) of untreated carcasses, 25/36 (69.4%) water treated, and 2/36 (5.6%) treated with PoultrypHresh™ were positive with Campylobacter (Figure 1). PoultrypHresh™ met FSIS standards for Campylobacter, which have a maximum acceptance of 15.7% in broiler carcasses. [4] These data demonstrate that an agitated dip treatment of PoultrypHresh™ significantly lowered the presence of Campylobacter on whole bird carcasses compared to either untreated or water treated carcasses (Table 1). Bird rinse samples directly plated with no enrichment demonstrated that 33.3% of untreated and 22.2% water treated carcasses were found to be positive for Campylobacter, while one of the 36 carcasses (2.8%) treated with PoultrypHresh™ at pH 1.4 were positive (Figure 2). Once rinsates were enriched using 2X Bolton’s broth, 75.0% of non-treated and 60.7% of water treated carcasses were contaminated with Campylobacter, while only 2.9% of carcasses treated with PoultrypHresh™ were contaminated (Figure 3).

The present study demonstrated that PoultrypHresh™ can significantly (P = 0.05) reduce the presence of natural Campylobacter on broiler carcasses. These data show that an antimicrobial dip can be effective in the elimination of Campylobacter and may be an option for lowering the prevalence in commercial processing facilities. PoultrypHresh™ applied as an agitated dip treatment for whole broiler carcasses at pH 1.4 for 20 s can significantly (P = 0.05) lower the number of Campylobacter positive broiler carcass rinse samples. A single hurdle application treatment of this antimicrobial may reduce the presence of Campylobacter on whole carcasses, helping poultry processors to succeed in meeting regulatory requirements. Additional PoultrypHresh™ applications at multiple intervention points throughout processing plants may further increase microbial reductions.

Table 1. Number of direct, enriched, and overall samples positive for Campylobacter in untreated, water treated, and PoultrypHresh™ treated at pH 1.4. Samples not positive on direct analysis were enriched and re-evaluated.







Pr > |t|1

































1 Probability that the treated groups are not different from the untreated control.

1 a,b values with different superscripts are significantly different by Chi Square test for independence.


Figure 1. Overall percentage of carcasses positive for Campylobacter in untreated, water treated, and PoultrypHresh™ treated at pH 1.41



2 a,b values with different superscripts are significantly different by Chi Square test for independence.


Figure 2. Percentage of Campylobacter in carcass rinses directly plated (no enrichment) of untreated, water treated, and PoultrypHresh™ at pH 1.4 treated carcasses in a 20 s agitated dip2



3 a,b values with different superscripts are significantly different by Chi Square test for independence.


Figure 3. Campylobacter presence in carcass rinse samples enriched for 48 h in 2X Bolton's Broth of untreated, water treated, and PoultrypHresh™ treatment at pH 1.43



Stricter performance standards by USDA-FSIS are forcing poultry processors to establish more effective means of treating foodborne microorganisms throughout plants. Campylobacter continues to be one of the main pathogens of concern for the poultry industry. The results showed a significant reduction in the presence of Campylobacter after carcasses were treated with PoultrypHresh™. This shows the product may be a valuable addition to processing plants and assist processors in meeting regulatory requirements.



1.       National Institute of Allergy and Infectious Diseases (NIH). Campylobacteriosis. United States Department of Health and Human Services. [Online] 2015 [cited 2018]. Available at: campylobacteriosis/Pages/default.aspx

2.       Guerrero-Legarreta I. Handbook of Poultry Science and Technology: Secondary Processing. John Wiley & Sons, Inc. 2010; 2:599-600.

3.       United States Department of Agriculture (USDA). USDA Finalizes New Food Safety Measures to Reduce Salmonella and Campylobacter in Poultry [Online] 2016 [cited 2018]. Available at: /02/0032. xml&printable=true&contentidonly=true

4.       Almanza AV. Changes to the Salmonella and Campylobacter Verification Testing Program. Food Safety and Inspection Service 2015; 80(16):3940-3950.

5.       Food Safety and Inspection Service (FSIS). New Performance Standards for Salmonella and Campylobacter in Not-Ready-to-Eat Comminuted Chicken and Turkey Products and Raw Chicken Parts and Changes to Related Agency Verification Procedures. United States Department of Agriculture, Federal Register 2016; 81(28):7285-7300.

6.       Dickens JA, Cox NA, Bailey JS, Thomson JE. Automated microbiological sampling of broiler carcasses. Poult Sci 1985; 64:1116-1120.

7.      MLG 41.04 United States Department of Agriculture. Laboratory Guidebook. Isolation and Identification of Campylobacter jejuni/coli/lari from Poultry Rinse, Sponge, and Raw Product Samples/ [Online] 2016 [cited 2018]. Available at: http://www.fsis.usda. gov/wps/wcm/connect /0273bc3d-2363-45b3-befb-1190c25f3c8b/MLG41.pdf? MOD=AJPERES

© 2016 The Author(s). This open access article is distributed under a Creative Commons Attribution (CC-BY) 4.0 license. You are free to: Share — copy and redistribute the material in any medium or format Adapt — remix, transform, and build upon the material for any purpose, even commercially. The licensor cannot revoke these freedoms as long as you follow the license terms. Under the following terms: Attribution — You must give appropriate credit, provide a link to the license, and indicate if changes were made. You may do so in any reasonable manner, but not in any way that suggests the licensor endorses you or your use. No additional restrictions You may not apply legal terms or technological measures that legally restrict others from doing anything the license permits
Editor in Chief
Prof. Dr. Asli UÇAR (PhD)
Department of Nutrition and Dietetics, Faculty of Health Sciences, University of Ankara, Ankara, Turkey


Prof. Dr. Asli UÇAR was awarded the PhD degree in Nutrition Sciences in 2006. She has excellent knowledge of food infection, hygiene of catering, attitudes of women towards food safety and Turkish perception of organic foods. Her major research area is food safety, nutrition science, food questionnaire and food hygiene. Prof. Asli UÇAR has recently completed three projects entitled Investigation of Effect of Some of Group B Vitamins and Vitamin D on Depression Level, Evaluation of Nutritional Status and Eating Habits of Children with Autism and Investigation of Effect Exposure to Steviol Glycoside on Oxidative Damage Index Parameters, Paraxonase Enzyme and Some Genetic Parameters in Balb/c Mice. Moreover, she partially contributed to compile the book (ISBN 978-953-51-2277-7) entitled Food Safety, Problems and Solutions - Significance, Prevention and Control of Food Related Diseases and wrote book chapter (ISBN 978-953-307-764-2) entitled Organic Food and Agriculture - New Trends and Developments in the Social Sciences. 

Journal Highlights
Key title: Advanced Food and Nutrition Sciences 
Abbreviation: Adv Food & Nutr Sci
Frequency: Annual

Current Volume: 2 (2017)
Next volume: 3 (2018)
Back volumes: 1-2
Starting year: 2016
Nature: Online
Submission: Online
Language: English


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