SALMONELLA SP. AND CALCIUM CONTENT AS ONE OF THE MOST IMPORTANT MICROBIOLOGICAL AND CHEMICAL QUALITY PARAMETERS OF MECHANICALLY SEPARATED MEAT

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3 Poster presentation SALMONELLA SP. AND CALCIUM CONTENT AS ONE OF THE MOST IMPORTANT MICROBIOLOGICAL AND CHEMICAL QUALITY PARAMETERS OF MECHANICALLY SEPARATED MEAT Babić J. 1, Aleksić-Agelidis A. 2, Jakšić S. 1, Kartalović B. 1, Živkov- Baloš M. 1, Petrović J. 1 Abstract According to the Regulation (EC) No 853/2004, Mechanically separated meat - MSM means the product obtained by removing meat from flesh-bearing bones after deboning or from poultry carcasses, using mechanical means resulting in the loss or modification of the muscle fiber structure. Concerning public health risks related to MSM, the aim of this paper was to investigate the most important microbiological and chemical quality parameters of mechanically separated meat samples from January 2015 to December 2015, such as Salmonella sp. and calcium content. In 5% of the samples the presence of Salmonella sp. species was revealed. S. Infantis and S. Enteritidis were isolated in 4 out of 11 Salmonella positive samples (36.36%) and S. Typhimurium was isolated in 3 out of 11 Salmonella positive samples (27.27%). The calcium content was examined in 38 samples and 28.95% of samples were within the concentration range of mg/kg, one of the samples had concentration even higher than 1500 mg/kg. According to our survey, the MSM is a food product with significantly high risk of the Salmonella presence and low compositional quality. Keywords: MSM, Salmonella, calcium Introduction Changes of the poultry processing industry have begun in the late 1950s and early 1960s (Owens et al., 2010). At that time as a result of an increase in the consumption of poultry meat, more parts such as frames, backs, necks, drumsticks, wings, etc. became available for mechanical separation (Froning and McKee, 2001). After the removal of the usual meat cuts, a certain amount of meat is always firmly attached to the bones. In the process of mechanical separation, that firmly attached meat is removed by grinding the starting material and passing it through a sieve under low or high pressure. Low pressure MSM has the EU upper limit of 100 mg/100 g (1000 ppm) calcium. MSM with calcium concentration above this threshold is considered to be high pressure MSM (EFSA, 2013). If calcium content does not exceed EU upper limit, MSM may be treated as minced meat, but any MSM additives to the final meat product have to be declared (Branscheid et al., 2008). 1 Babić Jelena, DVM, research assistant; Jakšić Sandra, PhD, research associate; Kartalović Brankica, specialist adviser; Živkov- Baloš Milica, PhD, senior research associate; Mihaljev Željko, PhD, research associate; Petrović Jelena, PhD, senior research associate, Scientific Veterinary Institute Novi Sad, Novi Sad, Serbia 2 Aleksic-Agelidis Aleksandra, DVM, University of Belgrade, Faculty of Veterinary Medicine, Belgrade, Serbia Corresponding author: Babić Jelena, jelenababic@niv.ns.ac.rs 552 P age

4 Until the end of last year the legislative act of the Republic of Serbia has identified two types of MSM - MSM produced using techniques that do not alter the structure of the bones with the upper limit of 100 mg/100 g and MSM produced using other techniques with the upper limit of 150 mg/100 g (Official Gazette RS, 31/2012). The legislative act the Republic of Serbia in force doesn t identify two types of MSM there is only one type with permitted level of 0.1% (100mg/100g) (Official Gazette RS, 94/2015). MSM contains muscle, fat and connective tissue like minced meat and meat preparations, but MSM can also contain fine bone particles and bone marrow (James et al., 2013). According to Regulation (EC) No 853/2004, Mechanically separated meat- MSM means the product obtained by removing meat from flesh-bearing bones after deboning or from poultry carcasses, using mechanical means resulting in the loss or modification of the muscle fiber structure. MSM, which is relatively low in fat and low in cost, is widely used to manufacture further processed products such as bologna, salami, frankfurters, turkey rolls, restructured meat products, and soup mixes (Froning and McKee, 2001; Nagy et al., 2007). Many researchers have remarked that MSM is an excellent medium for bacterial growth. MSM also presents a high potential health risk to the consumers because MSM is usually contaminated with microorganisms such as Salmonella sp. Campylobacter, Listeria sp. and Clostridium perfringens that originate from the carcass raw material. These pathogens usually multiply thanks to the inappropriate storage history and the processing environment, mainly as a result of poor hygienic measures (environment, handlers, and equipment) (ICMSF, 2005). The risk of the risk of microbial growth increases with the degree of muscle fiber degradation, thus high pressure MSM may provide a more favorable substrate for bacterial growth compared with low pressure MSM (EFSA, 2013). Also, improper holding temperatures during the production and storage phases allow growth and multiplication of contamination (Yuste et al., 2002). Under the food safety criteria, MSM must also be tested for Salmonella. Five samples of 10g each must be taken from one batch per sampling period and Salmonella must be absent in all samples. In order to control MSM microbiological quality USDA/FSIS has established the HACCP procedure (USDA/FSIS, 1999). Concerning public health risks related to MSM, the aim of this paper was to investigate the most important microbiological and chemical quality parameters of mechanically separated meat samples 553 P age Material and Methods From January to December 2015, a total of 49 samples of MSM were analyzed. Samples were transported to the laboratory after being collected in a portable cooler at a temperature of 4 C and microbiological analysis were carried out immediately. 36 samples of MSM were analyzed for Salmonella sp. presence. Concentration of calcium was measured in 38 samples of MSM. Isolation and identification of Salmonella sp. (in 10g of MSM) was performed according to standard methods (SRPS EN ISO 6579:2008). Calcium concentration was analyzed using flame emission spectrophotometry on flame photometer Jenway PFP7 (λ = 422,7 nm). Solution used for calibration was prepared from commercial stock standard solution

5 with 1000mg Ca/L (AccuTrace TM Reference Standard (USA)). Results and Discussion In 36 analyzed samples, Salmonella sp. was isolated from 11 samples or 30.6% from total number of analyzed samples. Three different serovars of Salmonella were found in 11 samples (Table 1.) Table 1. Salmonella sp. isolated from MSM samples. Salmonella Number of positive samples Percentage of total analyzed samples (%) Percentage of contaminated samples (%) S. Infantis S. Enteritidis S. Typhimurium Salmonella sp The calcium content was examined in 38 samples and 29% of samples were with the concentration range of mg/kg, one sample even had concentration higher than 150 mg/100 g (Table 2.) Table 2. Calcium content in analyzed MSM samples. Ca content (mg/kg) Number of MSM samples Percentage (%) Total Salmonellae are one of the most important food-borne pathogens. Owing to a high prevalence in livestock, they tend to spread through the food chain, exposing humans to the risk of salmonellosis. Salmonella sp. are among the major causes of foodborne disease in the U.S. and worldwide (Alali et al., 2016). In France, salmonellosis is the main cause of foodborne bacterial infection and the consumption of poultry products is considered as a major source of salmonellosis (Poirier et al., 2008). Seven Salmonella serotypes were identified from spleen and mechanically separated ground chicken and Salmonella prevalence in MSC samples was 27.8%. About 2% of MSC samples had relatively high Salmonella numbers (i.e., log MPN) which may pose a significant risk to human infection (Alali et al., 2016). Mattick and al. (2002) after investigation of the heat tolerance of S. Typhimurium in sausage-meat, concluded that Salmonella sp. are present in a significant proportion of sausages and are not always killed during the cooking process. Mechanically separated chicken meat is thought to be a major source of the contamination of sausage-meat with Salmonella. The products that are of a special 554 P age

6 concern are raw, smoked or lightly cooked products. Salmonella is usually killed by temperatures >50 C, with the death rate increasing as the temperature increases. As previously stated, the consumption of poultry products is a major source of Salmonella infections. Jovanović at al. (2013) at the Institute for hygiene and technology of meat, Belgrade, Serbia, investigated 152 samples of MSM for Salmonella contamination. Eight samples (5,26%) out of 152 were contaminated with Salmonella sp. The latest available results in Serbia have shown that the prevalence of Salmonella sp. from MSM is only 4% (Kureljušić at al., 2016) which is lower than our results. MSM has higher bone content and consequently calcium content as compared to fresh meat. The calcium content is frequently used as one of the criteria to identify MSM (Branscheid et al., 2009). The determination of bones (or calcium content) in MSM is a form of controlling the yield of mechanical separation processes. A high bone content means that the pressure used in the deboning process was too high or that the meat to bone ratio was too low (EFSA, 2013). Conclusion The MSM is food product with significantly high risk of the Salmonella presence and low chemical quality due to high percentage of chemical nonconformity results. Minimizing possibility for microbiological contamination and related hazards during production of MSM should be ultimate goal for poultry processing industry. That can be achieved through successfully implemented HACCP system built upon a solid foundation of prerequisite programs for good manufacturing practice and good hygiene practice. Holding proper temperature is also imperative during process of production, storage as well as transportation of MSM and products that contain MSM. References 1. Alali W.Q., Celik C., Wu D., Cui Y., Guran H. S., Mann D The relationship between Salmonella levels in chicken spleen and mechanically separated ground chicken. Food Control 66: Branscheid W., Judas M., Horeth R The morphological detection of bone and cartilage particles in mechanically separated meat. Meat Science 81: EFSA Scientific Opinion on the public health risks related to mechanically separated meat (MSM) derived from poultry and swine. EFSA Panel on Biological Hazards (BIOHAZ). European Food Safety Authority (EFSA), Parma, Italy. 11(3): Froning G.W. and McKee S.R., Mechanical separation of poultry meat and its use in products. In: Poultry meat processing. AR Sams. CRC Press, New York, ICMSF (International Commission on Microbiological Specifications for Foods) Microorganisms in Foods: Microbial Ecology of Food Commodities. 6. James C., Purnell, G., James, S.J Description of the Processes used in the UK to Manufacture MSM and Former DSM Meat Products from Poultry and Pork and an Initial Assessment of Microbiological Risk. A final report for the UK Food Standards Agency. FRPERC REF NO Jovanović J., Borović B., Velebit B., Lakićević B., Baltić T., Mitrović R., Milijašević M Mikrobiološki status pilećeg mehanički separisanog mesa. 555 Page

7 Tehnologija mesa 54 (2): Kureljušić J., Rokvić N., Karabasil N., Petrović J., Kureljušić B., Jezdimirović N Finding of Salmonella spp. in Mechanically Separated Meat. Second International Symposium of Veterinary Medicine- ISVM2016, Mattick K.L., Bailey R.A., Jørgensen F., Humphrey T.J Prevalence and number of Salmonella in sausages and their destruction by frying, grilling or barbecuing. Journal of applied microbiology. 93, Nagy J., Lenhardt L., Korimova L., Dičakova Z., Popelka P., Pipova M., Tomkova I Comparison of the quality of mechanically deboned poultry meat after different methods of separation. Meso. IX (2): Official Gazette RS No Rulebook no quality of minced meat, meat preparation and meat products 12. Official Gazette RS No Rulebook no quality of minced meat, meat preparation and meat products 13. Owens C. M., Alvarado C.Z., Sams A. R Poultry meat processing. CRC Press. 14. Poirier E., Watier L., Espie E., Weill F.-X., De Valk H., Desenclos J.-C Evaluation of the impact on human salmonellosis of control measures targeted to Salmonella Enteritidis and Typhimurium in poultry breeding using time-series analysis and intervention models in France. Epidemiology and Infection 136 (9): Regulation (EC) No 853/2004 of the European Parliament and of the Council of 29 April 2004 laying down specific hygiene rules for food of animal origin 16. USDA/FSIS (United States Department of Agriculture/Food Safety and Inspection Service) Generic HACCP Model for Mechanically Separated Species/ Mechanically Deboned Poultry. 17. Yuste J., Pla R., Capellas M., Mor-Mur M Application of high-pressure processing and nisin to mechanically recovered poultry meat for microbial decontamination. Food Control 13: P age