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Fate of Spoilage Microorganisms Associated with the Production of Pickled Sausage Using a Cold Fill Process

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Fate of Spoilage Microorganisms Associated with the Production of Pickled Sausage Using a Cold Fill Process PEER-REVIEWED ARTICLE Nelson J. Gaydos,1 Hassan Gourama,2 Food Protection Trends, Vol 36, No. 5, p. 348–356 Joshua A. Scheinberg,2 Catherine N. Cutter2 Copyright© 2016, International Association for Food Protection 1* 6200 Aurora Ave., Suite 200W, Des Moines, IA 50322-2864 and Jonathan A. Campbell 1Dept. of Animal Science, The Pennsylvania State University, 324 Henning Bldg., University Park, PA 16802, USA 2Dept. of Food Science, The Pennsylvania State University, 202 Food Science Bldg., University Park, PA 16802, USA Fate of Spoilage Microorganisms Associated with the Production of Pickled Sausage Using a Cold Fill Process ABSTRACT counts, 3.89 log10 CFU/g in 48 h for yeast counts, Pickling is a means of preserving a variety of and 4.09 log10 CFU/g in 24 h for mold counts. foods, including meat products. Cooked sausages To our knowledge, this experiment is the first to have traditionally been pickled using a heated demonstrate that cold fill pickling effectively reduces solution of vinegar, salt, and spices in a process and inhibits spoilage microorganisms. known as hot filling. However, hot fill pickling can result in quality defects. Alternatively, room INTRODUCTION temperature brine does not cause the various Pickling is a means of preservation that has been used for quality defects resulting from heated brine. To date, many years to extend and maintain the shelf life of various no study has determined the efficacy of a cold fill types of food products, such as fruits, vegetables, eggs, and pickling process to inhibit the growth of lactic acid meat. In particular, pickled sausage is a common acidified bacteria (LAB), yeasts, and molds associated with meat product that is generally prepared by curing and cooking acidified meat products. Therefore, the efficacy of sausages, which are then covered with a heated brine solution cold fill pickling of cooked sausages, using a brine containing an organic acid (typically acetic or citric acid), solution (5% acetic acid and 5% salt at 22–23ºC salt, spices, food coloring, and sometimes preservatives. and pH ~2.70), was validated for the reduction of The sausages are acidified or pickled in containers which are spoilage microorganisms by inoculating sausages sealed, letting the brine solution penetrate into the sausages, with select LAB (6.98 log10 CFU/g), yeasts (4.49 allowing the acidification of the meat product. The pickling log10 CFU/g), and molds (4.29 log10 CFU/g) process utilizes a combination of microbiological inhibitory over 28 days of pickling. The results revealed effects, which together prevent the growth and survival of reductions of 5.51 log10 CFU/g in 24 h for LAB microorganisms. Meat products may be preserved by several *Author for correspondence: Phone: +1 814.867.2880; Fax: +1 814.863.6042; E-mail: [email protected] 348 Food Protection Trends September/October methods, including placing product in an acidic environment Research on the microflora categorized as spoilage (reducing the pH), using salt to decrease the water activity microorganisms associated with acidified meat products (aw), and sealing the storage container to decrease the oxygen is limited. To our knowledge, no scientific literature exists level. Leistner and Gorris referred to the use of multiple that addresses either: (1) the microbial stability of pickled processing factors to inhibit microorganisms in food as “hurdle meat products or (2) the growth or survival of spoilage technology” (15). The use of multiple, sub-lethal factors to microorganisms associated with acidified meat and sausage kill and inhibit unwanted microorganisms works by disturbing products using a cold fill pickling process. Recent studies the homeostasis of microorganisms on multiple fronts have demonstrated the effectiveness of pickling processes simultaneously, resulting in the destruction of microorganisms using various organic acids to reduce the growth and and allowing for the preservation of the food product (13). survival of pathogenic bacteria in fruits and vegetables (1, Preventing the spoilage of such products is critical to ensure 13), cucumber purees, vegetable brines and pickles (5, 6, pickled sausages remain of good microbiological quality and 7) and hard cooked, pickled eggs (22, 26, 29). Research on are safe for consumption. pickled eggs may be most relevant to pickled meat products, The preparation of pickled sausage has traditionally used because of their similar protein content and susceptibility either a cold fill or hot fill process, in which cooked sausages to microbial contamination, although these studies did not are covered with either a cold or hot brine solution to begin address spoilage microorganisms such as LAB, yeasts, and the pickling process. However, hot filling has generally molds. Therefore, the purpose of this study was to validate resulted in significant quality defects, including cloudiness, the efficacy of a cold fill pickling process of cooked sausages turbidity, or discoloration of the brine, which make the to prevent the growth and survival of LAB, yeasts, and molds product less appealing to consumers. To avoid these quality associated with ready-to-eat (RTE) sausage products. defects, many commercial pickled sausage processors may utilize a cold fill process, in which the brine solution is kept at MATERIALS AND METHODS room temperature prior to pickling. In this study, a cooked, smoked, and cured sausage product The pickling of sausages has been regarded as a safe meth- was placed into an acidified brine (5% acetic acid (wt/vol); od of meat preservation by federal regulatory authorities. 5% salt (NaCl, wt/wt); 22–23ºC; pH ~2.70) solution and However, recent incidences of spoilage contamination involv- held at room temperature for 28 days. Table 1 highlights the ing pickled meat products have prompted the United States spoilage microorganisms chosen for the challenge study, Department of Agriculture, Food Safety and Inspection Ser- based on their prominence in fresh and RTE meat products, vice (USDA-FSIS) to reconsider the microbiological stability relevant isolation sources, and availability. of pickled sausages subjected to a cold fill pickling process (17). Current U.S. regulations addressing acidified foods Antagonism of select lactic acid bacteria and fungi (21 CFR parts 114 and 120) (32, 33) are mainly focused on Preliminary studies (data not shown) indicated that the canned food products intended to ensure processors properly lactic acid bacteria (LAB) cultures (Table 1) identified for address the potential growth of Clostridium botulinum. Title use in this challenge study inhibited selected mold cultures. 21 CFR part 114 (32) states that “…the pH must be main- All three LAB cultures inhibited both Penicillium nalgiovense tained at or below 4.6…” with no specification to the type or strains; Lactobacillus sakei ATCC 15521 inhibited both concentration of the acid, while title 21 CFR part 120 (33) Cladosporium cladosporioides strains; and L. curvatus 51436 states that “…manufacturers of acidified fruit and vegetable inhibited C. cladosporioides 11275. No inhibition occurred products must demonstrate that their process achieves a 5-log between the selected yeast and mold cultures. Therefore, the reduction in appropriate acid-tolerant pathogenic bacteria, inoculation and spoilage challenge study of sausages by fungi as validation of product safety.” It does not appear that either and LAB cultures were performed separately. of these policies has been applied to pickled meat products or used to address issues with spoilage. Current USDA-FSIS Preparation of selected lactic acid bacteria cultures and policies require processors of pickled meat products to challenges study inoculum demonstrate stability and safety by monitoring and recording LAB cultures were obtained from the American Type several quantitative parameters, including pH, aw, salt con- Culture Collection (ATCC; Manassas, VA). Lyophilized cul- centration, temperature and overall formulations. To date, no tures were transferred into fresh de Man, Rogosa and Sharpe specific scientific validation has been performed to demon- (MRS) broth (Difco, Sparks, MD). Both L. curvatus cultures strate which intrinsic and extrinsic factors are important to were incubated aerobically at 30ºC for 48 h, while L. sakei was ensure the stability and safety of pickled meat products. The incubated at 30ºC in a microaerophilic environment (5.0% lack of scientific validation has generated additional concerns O2, 10% CO2, 85% N2) in a CO2 incubator (VWR Interna- from Federal regulators about post-processing contamina- tional, West Chester, PA), following ATCC recommenda- tion, since it is unknown if a cold fill process might allow for tions. Colonies were then re-suspended in fresh MRS broth the survival and growth of spoilage microorganisms. containing 10% glycerol (VWR, Radnor, PA) and stored at September/October Food Protection Trends 349 Table 1. Spoilage microorganisms inoculated onto sausages prior to cold-fill pickling Spoilage Microorganism Genus/Species ATCC1/ID Isolation Source Lactobacillus curvatus 51436 fermented sausage Lactic Acid Bacteria Lactobacillus curvatus PTA5150 Kimchi (fermented Korean dish) (LAB) Lactobacillus sakei 15521 moto (sake starter culture) Debaryomyces hansenii 90624 salty pickle Debaryomyces hansenii 10619 horse meat sausage Yeast Candida zeylanoides 26318 Spanish sausage Candida zeylanoides 24745 grape must Cladosporium cladosporioides 60549 frozen lamb carcass Cladosporium cladosporioides 11275 wheat
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