Summary The investigation included fermented sausages produced in the countries of west and south-east Europe, Greece, Bosnia and Herzegovina, Croatia, Hungary, Italy and Serbia. The sausages were produced in local meat industries in a traditional way without the use of starter cultures. Samples were collected from three production batches on day 0 and again after 2, 4, 7, 14 and 28 days. Microbiological analyses included the principal ingredients (meat, fat tissue), casings and additives (sugar, mixture of spices, salt), and the finished products. From all three production batches of fermented sausages from each individual country, 150 strains of lactic acid bacteria and 150 strains of coagulase-negative cocci were isolated. Biochemical characteristics of the isolated microorganisms were determined by API system (bioMerieux), i.e. by API 50 CHL and API ® Staph. Identification of all strains was made using the computer program APILAB Plus. From the hygienic standpoint, it is highly important that Listeria monocytogenes, Salmonella spp. and Staphylococcus aureus were not found in the finished product. During all stages of investigation, lactobacilli and staphylococci prevailed.

A b s t r a c t:Within the project "Safety of traditional fermented sausages: Research on protective cultures and bacteriocins" the task of Work Package 5 (WP5) was pilot production of fermented sausages inoculated with pathogens using selected protective cultures or bacteriocins. Resulting from the previous work packages, three strains of Lactobacillus sakei (I151, I154 and I155) showed bacteriocin-based antimicrobial activity against Listeria monocytogenes only. Because of that, the Task 5.1. was slightly changed to production of fermented sausages inoculated with pathogen Listeria monocytogenes, together with the selected protective culture. These protective strains, as well as the strain of L. monocytogenes, were sent by the Italian partner and used for inoculation of sausages. For the purpose of this task, three batches of the indigenous sausages were prepared according to the recipes previously described in the 1st Annual Report. Within each batch, four groups of sausages were produced: a positive control group (sausages inoculated only with L. monocytogenes), and three groups of sausages inoculated with L. monocytogenes and one of the three protective strains of Lb. sakei (I151, I154 and I155). A slight advantage of the strain I151 over the other two was observed in antilisterial activity, which may suggest that the three investigated strains represented just three isolates of the same bacteria. This hypothesis was supported by the results of genetic investigation of bacteriocin determinants of the isolates carried out in WP3, where it was verified that all the Italian strains are subclones of the same population. Having in mind these results, it may be suggested to use only Lb. sakei I151 as a protective starter culture in further experimental investigation.

Within the project "Safety of traditional fermented sausages: Research on protective cultures and bacteriocins" the task of Work Package 5 (WP5) was pilot production of fermented sausages inoculated with pathogen using selected protective cultures or bacteriocins. Resulting from previous work packages, three strains of Lactobacillus sakei (I151, I154 and I155) showed bacteriocin-based antimicrobial activity against Listeria monocytogenes only. Because of that, the task 5.1. was slightly changed to production of fermented sausages inoculated with pathogen Listeria monocytogenes, together with the selected protective culture. Three protective strains, as well as the strain of L. monocytogenes, were sent by the Italian partner and used for inoculation of sausages. For the purpose of this task, three batches of the indigenous sausages were prepared according to the recipes previously described in the 1st Annual Report. Within each batch, four groups of sausages were produced: a positive control group (sausages inoculated only with L. monocytogenes), and one of the three protective strains of Lb. sakei (I151, I154 and I155). A slight advantage of the strain I151 over the other two was observed in antilisterial activity, which may suggest that three investigated starins represented just three isolates of the same bacteria. This hypothesis was supported by the results of genetic investigation of bacteriocin determinants of the isolates carried out in WP3, where it was verified that all the Italian strains are subclones of the same population. Having in mind these results, it may be suggested to use only Lb. sakei I151 as a protective starter culture in further experimental investigation.

Fermented sausages have a long tradition of production in certain regions of the world. For two years, research has been conducted under the EU sponsorship (FP5 "Safety of traditional fermented sausages: Research on protective cultures and bacteriocins") in the countries of west and southeast Europe (Greece, Bosnia and Herzegovina, Croatia, Hungary, Italy, Serbia and Montenegro). The first phase of research included the separation of lactobacilli and micrococci from the natural microflora of dry sausages during various stages of maturation. The investigation included fermented sausages produced in the countries of west and southeast Europe. The sausages were produced in local meat industries in a traditional way without the use of starter cultures. Their composition and parameters of maturation differed and were characteristic to the mode of production of traditional sausages in each member country of the research team. Samples were collected from three production batches on day 0 and again after 2, 4, 7, 14 and 28 days. Microbiological analysis included the principal ingredients (meat, fat tissue), casings and additives (sugar, spice mixture, salt), and finished products. It included the determination of total bacteria count, and the count of lactobacilli, micrococci, enterobacteria, enterococci, yeasts and moulds, in addition to the presence of S. aureus, L. monocytogenes and Salmonella spp. in 25 g of sample, and Pseudomonas spp., sulphite-reducing clostridia and the aerobic sporogenic bacteria count. From all three production batches of fermented sausages from each individual country were isolated 150 strains of lactobacilli and 150 strains of catalase-positive cocci, members of the family Micrococcaceae. Biochemical characteristics of the isolated microorganisms were determined by API-system (BioMerieux), i.e. by API 50 CHL and API® ; Staph. Identification of all strains was made using the computer program APILAB Plus. From the hygienic standpoint, it is highly important that L. monocytogenes, Salmonella spp. and S. Aureus were not found in the finished product. The number of other microorganisms in the course of fermentation, if present in the raw material or sausage stuffing, decreased. During all stages of investigation, lactobacilli and micrococci prevailed. These microorganisms are responsible for the formation of favourable organoleptic properties during maturation of dry sausages. Among the population of lactobacilli prevailed the strains Lb. sakei and Lb. curvatus. In addition, the following strains were isolated: Lb. plantarum, Lb. alimentarius, Lb. pentosus, Lb. rhamnosus, Lb. paracasei, Lb. brevis, Lb. plantarum/paraplantarum, Lb. fermentum, Lb. salivarius, Lb. bavaricus, Lb. acidophilus, Lb. maltoromicus, Lb. yamanashiensis, Lb. Delbrueckii, Lb. sanfrancisco, Lb. confosus, Lb. halotolerans, Lb. fructivorans, Lb. yamashienis, then Leuconostoc mesenteroides, Ln. paramesenteroides, Ln. lactis, Ln. citreum, Ln. oenos, and several strains of the genus Pediococcus. Most often, the isolated micrococci belonged to the genera Staphylococcus xylosus and S. saprophyticus. Furthermore, S. simulans, S. haemolyticus, S. caprae, S. capitis, S. sciuri, S. hominis, S. auricularis, S. warneri, S. cohni, S. epidermidis, S. carnosus, S. lentus and Micrococcus spp. were also isolated.

Investigations into the qualitative characteristics of kid goat meat were done on the meat of 44 crossbred kid goats of domestic Balkan White and Saanen goat at the age of 3–4 and 6–7 months. The results showed that the highest average percentage of water contained the meat of a male kid goat at the age of 6–7 months (76.45 %), while the lowest was in the meat of a male kid goat at the age of 3–4 months (70.73 %). The highest content of proteins was found in the meat of a male kid goat at the age of 6–7 months (21.28 %), while the lowest was in the meat of a female kid goat in younger investigated groups (19.14 %). The highest content of fats was in the meat of a female kid goat at the age of 3–4 months (5.53 %), while the lowest was in the meat of a male kid goat in the younger investigated group (1.21 %). Quantity of mineral matters in both age and sex groups ranged from 1.06 to 1.17 %. Due to its physical-chemical values, kid goat meat from both age and sex groups is considered a very valuable product in human nutrition.