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L.monocytogenes, fakultativni intracelularni patogen, je široko rasprostranjena u razliĉitoj sveţoj i preraĊenoj hrani. Ovaj patogen je uzroĉnik listerioze, retkog ali ozbiljnog oboljenja ljudi i ţivotinja, izazvanog konzumacijom kontaminirane hrane. L.monocytogenes moţe da preţivi i raste u širokom rasponu razliĉitih uslova sredine u kojoj se nalazi, bilo da je ona prirodna ili sredina modifikovana aktivnošću ĉoveka. Usled sposobnosti da preţivi i raste na niskim temperaturama, u sredinama sa visokom koncentracijom soli, smanjenom pH i niskim vrednostima aktivnosti vode, ĉest je stanovnik pogona za proizvodnju hrane. Zbog svoje sveprisutnosti u ţivotnoj sredini, kontrola pojave L.monocytogenes u pogonima za proizvodnju hrane predstavlja ozbiljan izazov za sistem bezbednosti hrane. U cilju unapreĊenja sistema bezbednosti hrane, uporedo sa razvojem legislative i standarda, razvijaju se i nove metode i tehnike identifikacije mikroorganizama. Sekvenciranje kompletnog genoma (eng. Whole genome sequencing - WGS) predstavlja ultimativnu metodu za ispitivanje prisustva patogena i otkrivanje njihovog porekla u hrani.Delikates proizvodi od mesa spremni za jelo, identifikovani su kao jedan od ĉestih izvora epidemija listerioze. Prisustvo L.monocyogenes u pogonu za proizvodnju tradicionalnih proizvoda od mesa u Crnoj Gori, ispitivano je tokom perioda od 2011. do 2014. godine. Za posmatrani period izolovano je 20 izolata L.monocytogenes iz proizvoda od mesa i pogona za njihovu proizvodnju koji su podvrgnuti molekularnoj karakterizaciji, serotipizaciji uz pomoć multipleks PCR-a i sekvencera nove generacije (eng. Next generation sequencing – NGS), utvrĊivanju potencijalne antimikrobne rezistencije na osnovu NGS podataka o prisustvu secifiĉnih gena rezistencije i skriningu na prisustvo odreĊenih markera rezistencije na dezinficijense..., L. monocytogenes, facultative intracellular pathogen, can be found in different types of raw and processed foods. This pathogen causes listeriosis, a rare but severe disease in humans and animals, which is spread primarily through the consumption of contaminated food. L. monocytogenes can survive and grow in a wide spectrum of environmental conditions, weather natural or habitats created by the human activity. Due to its ability to survive low temperatures, higher sodium nitrate concentrations, low pH and water activity, L. monocytogenes can frequently be found in the food processing environment. The ubiquitous nature of L. monocytogenes poses a significant encounter to its control in the food production environment, representing a great challenge for food safety. With the aim of improvement of the efficiency of the food safety systems, in parallel to the development of legislation and standards, the new methods and techniques for identification of microorganisms have been devised. Whole genome sequencing (WGS) technique represents the ultimate method for pathogen detection and for determination of their origin in food products.Ready-To-Eat (RTE) delicatessen meat products have repeatedly been identified as a source of listeriosis outbreaks. The presence of Listeria monocytogenes was evaluated in the meat processing facility in Montenegro in the period from 2011 to 2014. In total, 20 isolates of L. monocytogenes, isolated from traditional meat products and environmental swabs, were subjected to molecular characterization, serotyping by both multiplex PCR and next generation sequencing (NGS), testing to their potential antimicrobial resistance (AMR) by extraction of specific genes from NGS data and screening for the presence of some disinfectant resistance markers...

Meso i proizvodi od mesa, posebno meso živine, ima značajnu ulogu u pojavi kampilobakterioze kod ljudi koja je danas u svetu najčešća bolest prenosiva hranom. Brojni su izvori kontaminacije trupova živine u klanicama, uprkos nastojanjima i različitim merama da se učestalost kontaminacije smanji. Lečenje kampilobakterioze ljudi je otežano obzirom na sve veću učestalost rezistencije kampilobakterija na antimikrobne lekove. Cilj istraživanja u okviru ove doktorske disertacije bio je da se pomoću različitih tehnika izolacije i identifikacije (automatizovani kvalitativni test na principu ELFA tehnologije, selektivne podloge (mCCD agar i CampyFood agar), Real-time PCR tehnika (RTi-PCR) i Multipleks PCR tehnika (m-PCR)), utvrdi učestalost nalaza Campylobacter jejuni, Campylobacter coli i Campylobacter lari na ohlađenim trupovima živine iz tri objekta, ispita filogenetska srodnost Campylobacter jejuni, odnosno Campylobacter coli iz tri različita objekta i humanih izolata (dva izolata Campylobacter jejuni, odnosno tri izolata Campylobacter coli), kao i da se disk difuzionom metodom i E-testom utvrdi osetljivost na odabrane antimikrobne lekove. Korišćenjem automatizovanog kvalitativnog testa na principu ELFA (Enzyme Linked Fluorescent Assay) tehnologije Campylobacter spp. su dokazane kod 91,30% trupova (ispirak ohlađenih trupova) živine poreklom iz dve industrijske i jedne zanatske klanice (objekata „A“, „B“ i „C“). Zasejavanjem uzoraka ispiraka ohlađenih trupova živine iz sve tri klanice na selektivni modifikovani ugljeni cefperazon deoksiholat agar (mCCD) i CampyFood agar, prisustvo Campylobacter spp. utvrđeno je kod 91,30%, odnosno 77,39% trupova živine. Pomoću RealTime PCR tehnike (16S rRNK) na trupovima živine iz sva tri objekta prisustvo Campylobacter spp. utvrđeno je u 77,39% uzoraka. U uzorcima ispiraka ohlađenih trupova živine iz zanatske klanice (objekat „C“) učestalost nalaza Campylobacter spp. bila je statistički značajno veća (p < 0,05) u odnosu na učestalost nal, Meat and meat products, especially poultry meat, play a significant role in the occurrence of campylobacteriosis in humans, which is the most common food-borne disease in the world today. There are numerous sources of contamination of poultry carcasses in slaughterhouses, despite efforts and various measures taken in order to reduce the frequency of contamination. The treatment of campylobacteriosis in humans is difficult due to the increasing frequency of resistance of Campylobacter to antimicrobial drugs. The aim of the research in this doctoral dissertation was to use different isolation and identification techniques (automated qualitative test on the principle of ELFA technology, selective substrate (mCCD agar and CampyFood agar), Real-time PCR technique (RTi-PCR) and Multiplex PCR technique (m-PCR)), in order to determine the frequency of Campylobacter jejuni, Campylobacter coli and Campylobacter lari on chilled poultry carcasses from three facilities, examined the phylogenetic relationship of Campylobacter jejuni and Campylobacter coli from three different facilities and human isolates (two isolates of Campylobacter jejuni and three Campylobacter coli isolate), as well as to determine the sensitivity to selected antimicrobial drugs by disk diffusion method and E-test. Using an automated qualitative test on the principle of ELFA (Enzyme Linked Fluorescent Assay) technology Campylobacter spp. were detected in 91.30% of poultry carcasses (whole-carcass rince) originating from two industrial and one artisanal slaughterhouses (facilities "A", "B" and "C"). Inoculation of the rinses from chilled poultry carcasses from all three slaughterhouses on selective modified carbon cefperazone deoxycholate agar (mCCD) and CampyFood agar, showed the prevalence of Campylobacter spp. was 91.30% and 77.39%, respectively. Using RealTime PCR technique (16S rRNA) on samples from poultry carcasses from all three investigated facilities, the prevalence of Campylobacter spp. was 77.39%

Индустрију меса карактерише велика потрошња воде и неоспорно је да представља веома важан извор органског загађења животне средине. За пречишћавање отпадних вода постоје еколошки као и финансијски мотиви који се међусобно преплићу и допуњују. Коришћење доступних технологија за гајење рибa у склопу кланичне индустрије представља еколошко решење уклањања органски оптерећене воде искоришћене у кланици. Са друге стране, овакав интегрисани систем носи и одређене ризике који се углавном односе на безбедност меса рибе произведене на овај начин. Циљ истраживања које је спроведено у оквиру ове докторске дисертације је да се испита утицај коришћења отпадних вода из кланице на здравље шарана и добијање меса шарана безбедног за исхрану људи. Истраживање у оквиру докторске дисертације је обављено у више фаза. Изградња система за пречишћавање отпадне воде као дела кланице спроведена је у оквиру Индустрије меса „Ђурђевић“ у Пећинцима. Испитана је ефикасност рада пречистача применом хемијских анализа воде пре и после пречишћавања воде. У оквиру кланице је затим изграђен рибњачки објекат који се већим делом снабдева водом из система за пречишћавање отпадне воде из кланице. Вода из пречистача одлази у предрибњак где се врши аерација пречишћене воде, након чега вода улази у рибњак у којем се део нутријената из пречистача искоришћава за исхрану шарана. Вода се након тога користи за наводњавање земље која се налази у околини кланице. На овај начин рибњак има утицај на повећање квалитета отпадних вода и достизање граничних вредности које су прописане пре уливања у природни реципијент. Насађени су младунци шарана добре кондиције и здравственог стања. Производња шарана се одвијала у оптималним амбијенталним условима употребом пречишћене воде пореклом из кланице уз додатак бунарске воде..., Meat industry is characterised by high water consumption and is undoubtedly a significant source of organic pollution of the environment. There are both ecological and financial motives for water purification which intertwine and complement each other. The use of available technology for fish farming in slaughterhouse industry is an ecological solution for removal of organically loaded water used in slaughterhouses. On the other hand, such integrated system carries certain risks mostly related to the safety of fish meat produced in this way. The aim of the research carried out in this doctoral thesis was to examine the effect of the use of wastewater from slaughterhouses on the health of carp and safety of carp meat for human consumption. The research of this doctoral thesis was carried out in several stages. A system for wastewater purification was built on the property of ‘Djurdjević’ Meat Industry in Pećinci. The efficiency of wastewater treatment plant was examined by using chemical analyses of water in different water purifying stages. After this, a fishpond was built on the same property. It was mostly supplied with the water from the system for wastewater purification from the slaughterhouse. The water from purifiers goes to pre-fishery where purified water is aerated, after which the water goes into fishpond where a part of nutrients from purifiers is used for carp nutrition. The water is then used for irrigation of the soil surrounding the slaughterhouse. This way, the fishpond has an impact on increasing the quality of purified wastewaters and reaching limit concentrations set before the inflow into a natural recipient. Carp fingerlings in good health were stocked in the fishpond. Carp was produced in optimal ambient conditions by using purified water originating from slaughterhouses and by adding well water. A good quality of meat was achieved by feeding fish balanced meals...

Cilj ispitivanja u okviru ove doktorske disertacije odnosio se na mogućnost inaktivacije Listeria monocytogenes u fermentisanim kobasicama. Za potrebe ispitivanja proizvedene su kobasice kontaminirane sa L. monocytogenes, sa i bez dodate starter kulture. Kontrolne grupe kobasica nisu bile kontaminirane. U toku zrenja praćene su promene mikrobiološkog statusa fermentisanih kobasica i fizičko-hemijskih osobina. Za sva ispitivanja korišćene su standardne metode. U fermentisanim kobasicama užeg i šireg dijametra sa i bez dodate starter kulture u toku zrenja broj bakterija L. monocytogenes se smanjivao se posle trećeg dana zrenja. U uzorcima fermentisanih kobasica užeg dijametra, sa i bez dodate starter kulture, na kraju procesa zrenja (18. dan) nije utvrđeno prisustvo bakterija L. monocytogenes. U uzorcima fermentisanih kobasica šireg dijametra sa dodatom starter kulturom, L. monocytogenes nije izolovana 21. dana zrenja, a bez dodate starter kulture od 31. dana zrenja. Prosečan broj enterobakterija u uzorcima fermentisanih kobasica užeg dijametra, sa i bez dodate starter kulture smanjivao se od trećeg dana zrenja, a kod fermentisanih kobasica šireg dijametra od sedmog dana zrenja. Prisustvo enterobakterija nije utvrđeno u uzorcima kobasica užeg dijametra na kraju procesa zrenja (18. dan), a u uzorcima kobasica šireg dijametra 31. dana zrenja. Tokom svih dana ispitivanja broj bakterija mlečne kiseline (BMK) u fermentisanim kobasicama užeg, odnosno šireg dijametra bio je veći od broja u kobasicama bez dodate starter kulture. Broj BMK rastao je kod kobasica užeg dijametra do 14. dana, a zatim je opadao do 18. dana, dok je kod fermentisanih kobasica šireg dijametra rastao do 31. dana, a zatim opadao do 41. dana zrenja. Ukupan broj aerobnih mezofilnih bakterija u fermentisanim kobasicama užeg, odnosno šireg dijametra rastao je do 14., odnosno 31. dana zrenja, a zatim je do kraja zrenja opadao. Između ukupnog broja aerobnih mezofilnih bakterija u uzorcima fermentisanih kobasi, The aim of the study under this doctoral dissertation was to address the possibility of inactivation of Listeria monocytogenes in fermented sausages. For testing purposes, sausages contaminated with L. monocytogenes were produced, with and without the starter culture added. The sausage control groups were not contaminated. Changes in the microbiological status of fermented sausages and physicochemical properties were monitored during ripening. Standard methods were used for all tests. In fermented sausages of narrower and wider diameter with and without added starter culture during ripening, L. monocytogenes decreased after the third day of ripening. In samples of fermented short-diameter sausages, with and without added starter culture, no L. monocytogenes was detected at the end of the ripening process (day 18). In specimens of broad-diameter fermented sausages with added starter culture, L. monocytogenes was not isolated on day 21 of ripening, and without added starter culture from day 31 of ripening. The average number of enterobacteria in narrower diameter fermented sausage samples, with and without added starter culture, decreased from the third day of ripening, and in fermented sausages of wider diameter from the seventh day of ripening. The presence of enterobacteria was not detected in narrow-diameter sausage specimens at the end of the ripening process (day 18) and in broad-diameter sausage samples on the 31st day of ripening. During all test days, the number of lactic acid bacteria (LAB) in the fermented sausages of narrower or wider diameter was greater than the number in sausages without added starter culture. LAB number increased in narrower diameter sausages by day 14, then decreased by day 18, whereas in fermented sausages of wider diameter, it increased by day 31 and then decreased by day 41. The total number of aerobic mesophilic bacteria in the fermented sausages of narrower or wider diameter increased to the 14th and 31st day of ripening, and the

The most common types of beta-casein in cow’s milk are A1 and A2, which differ in one amino acid. During the gastrointestinal proteolysis of A1 beta-casein in humans this difference results in the release of beta-casomorphin-7, an opioid which may lead to severe effects on human health, causing various ailments (type-1 diabetes mellitus, ischemic heart disease, arteriosclerosis, sudden infant death syndrome, autism, schizophrenia, gastrointestinal digestive discomfort, as well as increased gastrointestinal transit time). By contrast, A2 beta-casein cannot exert these effects owing to its different composition and metabolism. Furthermore, studies have shown that it can influence milk productivity traits. Our research aimed to screen the frequency of A1 and A2 alleles of beta-casein gene in a population of Serbian Holstein-Friesian cows and to detect how the genotypes influence milk production, and milk protein and fat yields. Out of 106 animals, 13 (12.26%) were of A1A1 genotype, 58 (54.72%) of A1A2, and 35 (33.02%) of A2A2 genotype. Milk yield was significantly (P<0.01) higher in A2A2 compared to both A1A1 and A1A2 genotypes. Milk protein concentrations were significantly (P<0.01) higher in A2A2 compared to A1A2 genotype, while milk fat concentrations were significantly (P<0.01) higher in A2A2 compared to both A1A1 and A1A2 genotypes., Najzastupljeniji tipovi beta kazeina u kravljem mleku su A1 i A2 koji se razlikuju u jednoj aminokiselini. Tokom gastrointestinalne proteolize A1 beta kazeina kod ljudi, oslobađa se betakazomorfin 7, opioidni peptid koji može uzrokovati pojavu raznih oboljenja (dijabetes melitus tip 1, ishemijska bolest srca, ateroskleroza, sindrom iznenadne smrti odojčadi, autizam, šizofrenija, nelagodnost u digestivnom sistemu, kao i produženo vreme gastrointestinalne pasaže). Suprotno tome, A2 beta kazein, zbog svog sastava i drugačijeg metabolizma, ne može da ispolji navedene efekte. Istraživanja su pokazala takođe, da može imati uticaj na proizvodnju i sastav mleka. Ovo istraživanje imalo je za cilj da ispita frekvenciju beta kazeinskih genskih alela A1 i A2 u populaciji holštajnfrizijske rase u Srbiji i da utvrdi kako genotipovi utiču na proizvodnju mleka, mlečnih proteina i masti. Od 106 ispitanih životinja, kod 13 (12,26%) je ustanovljen genotip A1A1, kod 58 (54,72%) A1A2, a kod 35 (33,02%) A2A2 genotip. Proizvodnja mleka bila je značajno veća kod životinja sa A2A2 genotipom u poređenju sa genotipovima A1A1 i A1A2 (P<0,01). Koncentracije proteina u mleku bile su značajno veće kod životinja sa A2A2 genotipom u poređenju sa životinjama A1A2 genotipa (P<0,01), dok su koncentracije mlečne masti bile značajno veće kod životinja A2A2 genotip u poređenju sa životinjama A1A1 i A1A2 genotipa (P<0,01).

This study reports, for the first time, different physico-chemical analyses, of Mediterranean mussels (Mytilus galloprovincialis) from harvesting areas in the Montenegro coast of the Adriatic Sea, in order to evaluate the influence of origin on different parameters and assessed the quality of shellfish grown in this area. The Boka Kotorska Bay is situated in Montenegro, in the southeastern part of the Adriatic Sea The mussel samples were collected at the same time in the spring of 2019 at six locations in Boka Kotorska Bay, Montenegro: Kotor and Tivat Bays. Biomet-ric parameters, percentage of meat, condition index, proximate composition, sensory evaluation and lipid profiles of mussels were studied. The concentrations of some micro and macro elements and heavy metals in mussels, were also analyzed. Significant differences were found between mussels from different locations. Mean biometric parameters of mussels grown in Sv.Nedjelja, were considerably higher than in mussels grown elsewhere.. Protein, lipid, ash and gly-cogen content were varied from 7.80 to 10.26%; 1.36 to 2.18%; 1.73 to 3.34% and 12.81 to 15.38%, respectively. Gas chromatographic analysis showed that polyunsaturated fatty acids (PUFAs) were dominant lipids in mussels (37.56 to 41.08%), followed by monounsaturated (MUFAs) (30.52 to 38.31%) and saturated (SFAs) (21.89 to 29.45%) fatty acids. Fatty acid profiles were investigated and revealed high contents of n-3 PUFAs and high n-3/n-6 ratios in all mussels from Montenegro In the mussel samples high concentrations of K, Mg, Ca, and Na, and much lower concentrations of Fe, Mn, Zn, and Cu were found. Some of toxic elements (As, Pb, Hg and Cd) were determined also.. The qualitative sensory assessment showed that all mussels were acceptable. All mussels in the current study achieved scores of 3 or more out of 5 in the qualitative category. Data on biochemical composition and quality indices of the mussels cultured in the Boka Kotorska Bay demonstrated that

Sremski kulen is a wide diameter dry fermented sausage, produced from pork, seasoned with red spicy paprika, stuffed into pork cecum, and preserved by smoking, fermentation and drying. Due to specific ripening process, Sremski kulen is suitable for the accumulation of biogenic amines. Therefore, the aminogenesis was studied in traditionally produced Sremski kulen, taking into account the physicochemical parameters and microbial counts. The content of six biogenic amines (tryptamine, phenylethylamine, tyramine, histamine, putrescine, and cadaverine) was analyzed using high-performance liquid chromatography. The ripening process of Sremski kulen was slow followed by changes in aw and pH value as well as expressed proteolysis. The autochthonous microbiota showed pronounced decarboxylase activity. Tryptamine and phenylethylamine were detected at each examined ripening stage while histamine was not detected until the end of ripening (16.55 ± 2.33 mg/kg). Tyramine, cadaverine, and putrescine content significantly increased during the ripening period (p < .05). In the final product, cadaverine was the dominant biogenic amine (132.40 ± 5.05 mg/kg), followed by tyramine (115.80 ± 15.46 mg/kg) and putrescine (68.55 ± 2.39 mg/kg). Although the long ripening period greatly contributed to the accumulation of biogenic amines in final product, their content are not of concern from product safety aspects, but requires improvement in hygiene of production process.

The fatty acid (FA) profiles of lamb muscle and adipose tissue originating from ten different grazing areas in North Macedonia and an assessment of whether the meat origin could be distinguished on the basis of FA profile are presented. Muscle and adipose tissues of three-month-old male lambs (crossbreds of autochthonous Ovcepolian and Wurttemberg) reared on spring pastures were studied. Statistically significant differences in the individual FA contents, FA groups and FA ratios were observed between lamb tissues from the different geographic regions. Canonical discriminant analysis showed there was a significant linear divergence between tissues from almost all examined regions. The greatest weight in the differentiation of the different areas showed fatty acid ratios, C18:3n3, C18:1n9c and C20:5n3 for the muscle tissue, as well as C18:1n9c, C18:2n6c, C16:1 and C17:1 for the adipose tissue. This showed that the FA composition of muscle and adipose tissue obtained from lambs reared on pasture could serve as a useful indicator to aid differentiation of its geographic origin but it should be confirmed through further replicated studies.

Proliferative enteropathy, also known as proliferative ileitis, is considered to be one of the most economically important diseases in pig production worldwide. The estimated losses per affected growing pig usually range from US $1 to $5. The disease is caused by Lawsonia intrawellularis, a Gram-negative, obligately intracellular bacterium. Control of the disease can be achieved with the use of vaccines or antibiotics. Recently there has been an increase in the efforts in the control of certain pathologies of the digestive system with phytogenic additives. The aim of this work was to assess the effects of a phytogenic additive on the histomorphometric characteristics of the intestines in weaned pigs with a subclinical infection with L. intracellularis acquired spontaneously. Histomorphometry analysis showed that crypt depth was significantly shorter (P <0.05), and the villus-height-to-crypt-depth ratio (P<0.05) significantly greater in the treatment group than the control. This improvement in the histological parameters of the intestine, considered to be indicators of its health, proved the positive effect of the tested additive on the digestive system in pigs., Proliferativna enteropatija poznata kao i proliferativni ileitis smatra se jednom od ekonomski najznačajnih bolesti u svinjarskoj proizvodnji širom sveta. Procenjeni gubici po obolelom tovljeniku obično se kreću od 1 do 5 američkih dolara. Uzročnik ovog oboljenja je Lawsonia intracellularis, gram negativna obligatna intracelularna bakterija. Kontrola ovog oboljenja ostvaruje se primenom vakcinacije ili antibiotika. U poslednje vreme sve je češća primena fi togenih aditiva u cilju kontolisanja određenih patoloških stanja digestivnog trakta. Cilj ovog rada bio je da se ustanove efekti fi togenog aditiva na histomorfometrijske karakteristike creva kod odlučenih prasadi supklinički prirodno infi ciranih L. intracellularis. Histomorfometrijska analiza pokazala je da su kripte bile značajno (P<0,05) pliće, a količnik visine vilusa i dubine kripti značajno veći (P<0,05) u tretiranoj grupi prasadi nego u kontroli. Ovo poboljšanje histoloških parametara creva, koje se smatraju indikatorima njegovog zdravlja, dokaz je pozitivnog efekta ispitivanog aditiva na digestivni sistem svinja.

The main objective of the present research was to determine the concentrations of the selected pesticides in muscle, liver and skin of common carp. Fish were sampled in two different seasons from fish pond which received previously treated slaughterhouse wastewater. Pesticides including etridiazole, chloroneb, trifluralin, propachlor, chlorothalonil, hexa-chlorocyclopentadiene, atrazine, simazine, alachlor, metribuzin, metolachlor, DCPA, cyanazine, chlorobenzilate, endrin aldehyde, cis permethrin and trans permethrin were determined by using a GS-MS method. Many of pesticides were not determined or determined in low concentrations. Propachlor was found in muscle, skin and liver. The recommended acceptable daily intake was higher in comparison with the estimated daily intake for examined pesticides via fish reared in treated slaughterhouse wastewater. It is very important to maintain the safety of the fresh fish produced in wastewater in order to ensure food safety and avoid health problems in humans.

Izolacija i identifikacija Salmonella spp. ostaje nezamenjiva kao potvrdna metoda u mikrobiologiji hrane i hrane za životinje. Međutim, savremena industrija hrane, ali i javno zdravstvo zahtevaju razvoj novih, brzih metoda za detekciju Salmonella spp., jednog od najvažnijih patogenih mikroorganizama prenosivih hranom. Pored toga, tokom epidemioloških studija, uglavnom nije dovoljno da se izolati Salmonella tipiziraju do nivoa vrste i serotipa, već je neophodna i primena metoda tipizacije koje mogu napraviti razliku između epidemiološki različitih, ali genetski srodnih izolata. Veliki broj alternativnih metoda, uključujući i real-time PCR protokole, razvijen je i validovan u cilju detekcije Salmonella spp. Pored toga, napredak u razvoju molekularnih metoda pružio je alate visoke diskriminatorne moći koji su posebno mesto primene zauzeli pri određivanju izvora kontaminacije hrane, omogućavajući brzu i izuzetno pouzdanu tipizaciju Salmonella spp. Ova doktorska disertacija obuhvata četiri osnovne celine: (1)“In house” validacija real-time PCR protokola za detekciju invA i ttr gena Salmonella spp. u hrani i hrani za životinje; (2) „In house“ validacija real-time PCR protokola za detekciju Salmonella Enteritidis i S. Typhimurium u hrani; (3) Razvoj i „in house“ validacija novog real-time PCR protokola za detekciju invA gena Salmonella spp. u hrani; (4) Molekularna karakterizacija i ispitivanje antimikrobne osetljivosti Salmonella Enteritidis and S. Typhimurium izolovanih iz hrane, hrane za životinje i fecesa. Tokom izrade ove doktorske disertacije formirana je kolekcija od 60 izolata Salmonella Enteritidis i 60 izolata S. Typhimurium (po 12 poreklom ljudi, 24 poreklom iz hrane i 24 poreklom iz fecesa živine), a zatim je izvršena njihova molekularna karakterizacija i ispitivanje antimikrobne osetljivosti. Ispitivanje antimikrobne osetljivosti Salmonella izvršeno je disk difuzionom metodom prema EUCAST protokolu, dok je za određivanje minimalne inhibitorne koncentracije pri, method in the microbiology of food and feed. However, the modern food industry and public health require the development of new, rapid methods for the detection of Salmonella spp., one of the most important foodborne pathogens. In addition, during epidemiological studies, it is generally not sufficient to determine Salmonella isolates up to the species or the serotype level, but it is also necessary to apply subtyping methods that can distinguish between epidemiologically distinct, but genetically related isolates. A number of alternative methods, including real-time PCR protocols, have been developed and validated for the detection of Salmonella spp. In addition, the advances in the development of molecular methods have provided tools of high discriminatory power that have been particularly useful in determining food contamination sources, enabling fast and highly reliable typing of Salmonella spp. This dissertation has four major sections: (1) In-house validation of the real-time PCR protocol for the detection of invA and ttr genes of Salmonella spp. in food and feed; (2) In-house validation of the real-time PCR protocol for the detection of Salmonella Enteritidis and S. Typhimurium in food; (3) Development and in-house validation of new real-time PCR protocol for the detection of the invA gene of Salmonella spp. in food; (4) Molecular characterization and antimicrobial susceptibility testing of Salmonella Enteritidis and S. Typhimurium isolated from food, feed and feces. An extensive validation study, where real-time PCR protocols for the detection of invA and ttr of Salmonella spp. in food and feed were tested, gave excellent results and showed that they can be used as an adequate replacement for the standard method. By comparing different DNA extraction procedures, lower Ct values were achieved by using "Chelex extraction" rather than by extraction based on the thermal lysis of the cell...

Human trichinellosis and Trichinella infection in pigs are both still endemic in the Balkans, including Serbia. Because of the flow between the sylvatic and the domestic cycle of Trichinella spp., monitoring wildlife has been recommended for the risk assessment of Trichinella spp. infection in swine. We have previously shown the presence of Trichinella infection in wild carnivores including the wolf and the golden jackal, and here we report on Trichinella infection in several other mesocarnivore species. From a total of 469 animals collected between 1994 and 2013, Trichinella larvae were detected in 29 (6.2%, 95% CI = 4.0-8.4) animals, including 14 red foxes (4.7%), 7 wild cats (35%), 5 beech martens (4.8%), 2 pine martens (16.7%), and 1 European badger (6.25%). No Trichinella larvae were detected in the examined specimens of European polecats, steppe polecats and European otters. Species identification of the Trichinella larvae performed for 18 positive samples revealed T. spiralis in 77.8% and T. britovi in 22.2% of the isolates. Both species were detected in red foxes and wild cats. The predominance of T. spiralis in wildlife in Serbia indicates the (past or present) spillover of this pathogen from domestic to wild animals.

The aim of this study was to determine the content of some elements (Pb, Cd, Hg, Fe, Cu, Zn, As) in muscle tissue of Prussian carp (Carassius auratus gibelio) from seven different lakes in the Belgrade region, Serbia. Concentrations of Pb, Hg and As in fish muscle tissues from all examined lakes were under the maximum residual levels prescribed by the European Union (EU) and the maximum allowed concentrations (MAC) for Serbia. In all investigated fish, levels of Cd exceeded maximum allowed concentrations (0.05 mg kg-1). Data on the finding of elements in fish speak concurrently about the safety of fish as food and are good indicators of environmental pollution.

Poor animal welfare is a growing issue of concern all over the world. During pre-slaughter phases, some activities, if poorly performed, lead to stress reactions in animals. Animals responses to a great extent depend on species, breed, sex and age of animal. Food and water deprivation, exposure to unfavourable environmental conditions, improper handling and social mixing are just some indicators of compromised animal well-being. All these detrimental activities lead to the development of carcass and meat quality defects. Among them, important defects are untimely death of animals, body weight loss, carcass damage, and the two major meat quality defects, i.e. pale, soft, exudative (PSE) and dark, firm, dry (DFD) meats. Consumer dissatisfaction results, and with economic and financial losses, producers are placed in a disadvantageous position.

The presence of Listeria monocytogenes was evaluated in a small-scale meat processing facility in Montenegro during 2011-2014. L. monocytogenes isolates from traditional meat products and environmental swabs were subjected to a) molecular characterization b) serotyping by both multiplex PCR and next generation sequencing (NGS) c) potential antimicrobial resistance (AMR) was assessed by extraction of specific genes from NGS data and d) screening for the presence of some disinfectant resistance markers. Overall, traditional meat products were contaminated, most likely from incoming raw materials, with 4 major specific STs of L. monocytogenes (ST515, ST8, ST21, ST121) representing 4 clonal complexes (CC1, CC8, CC21, CC121) identified during the four-year period. These strains belonged to serogroup IIa which predominated, followed by IVb (ST515, CC1). The strains from environmental swabs belonged, exclusively, to ST21 and were isolated from cutting board and floor swabs in 2011. Furthermore, we found Tn6188, a novel transposon conferring tolerance to BC, to be specific to sequence type ST121. In addition, antimicrobial resistance genes mprF and fosX were present in clonal complexes CC21 and CC121, while complexes CC8 and CC1 exclusively harbored the mprF antimicrobial resistance gene.

Meat technology includes all steps from animal handling and slaughtering to production of different meat products. During animal handling, special attention is paid to animal welfare, not only to protect animals from suffering but also because of animal welfares importance for meat quality. The oldest processing methods (chilling, freezing, salting, smoking and drying) are being readjusted with respect to equipment and consumers health issues. Special attention is given to preservation of meats nutritive value (milder heat treatment) and health promoting properties of the products (functional foods). Novel methods (irradiation, high pressure, pulsed electric field, pulsed light and cold plasma) struggle with some issues such as acceptance by consumers, expensive equipment and effects on foods sensory properties. Along with novel products, demand for traditional meat products is still increasing which requires the the uniqueness and quality of these products to be preserved, along with increased production capacity.

Introduction. Trichinellosis is a cosmopolitan anthropozoonosis, and is a group of systemic diseases caused by larval forms of Trichinella. This is one of the oldest and most controversial parasitic zoonoses that has been in the forefront of veterinary, medical and biological research for many years. Parasites from the genus Trichinella were diagnosed in more than one hundred species of domestic and wild mammals and birds. The aim of this study was economic evaluation of a program for monitoring and controlling Trichinella in pigs. Materials and Methods. A systematic pest rodent control program deemed suitable for monitoring and controlling Trichinella in pigs was implemented in a representative test zone. The results of the pest rodent control program were analyzed using cost-benefit analysis to determine if a similar program would be suitable for a larger land area. Results and Conclusions. Prior to pest control, the incidence of Trichinella in pigs living in the test zone increased annually, while after systematic pest rodent control, the incidence of Trichinella-positive pigs decreased. These results, as well as relevant economic indicators, enabled us to design two mathematical models describing the control and monitoring of Trichinella in a larger land area. In this area, Srem Region, Serbia, a suitable Trichinella control program in pigs is clearly justified because the profit to cost ratio was positive for both models., Uvod. Trihineloza je kosmopolitska antropozoonoza, koja se ubraja u grupu sistemskih bolesti, a prouzrokovana je larvenim oblicima valjkastog crva Trichinella species. Ona je jedna od najstarijih i najupornijih parazitskih zoonoza koja se već duži niz godina nalazi u žiži interesovanja veterinarskih, medicinskih i bioloških naučno-istraživačkih radnika. Paraziti iz roda Trichinella dijagnostikovani su u više od sto vrsta domaćih i divljih sisara i ptica. Postavljeni cilj ovog istraživanja je procena efikasnosti programa za prac'enje i iskorenjivanje trihineloze pomoću analize dobiti i troškova. Materijal i metode. Procena efikasnosti programa za prac'enje i iskorenjivanje trihineloze urađena je kroz analizu dobiti i troškova U jednom naseljenom mestu izvedena je kontrolisana sistemska deratizacija. Rezultati i zaključak. Pre deratizacije pojava trihineloze u naseljenom mestu imala je pravolinijsku uzlaznu tendenciju, dok je nakon sistematske deratizacije tendencija pojave pozitivnih slučajeva bila negativna. Rezultati dobijeni na ovaj način, kao i relevantni ekonomski pokazatelji, omoguc'ili su NAM da dizajniramo dva programska modela za kontrolu, prac'enje i iskorenjivanje ove parazitoze na ovom području. Na osnovu analize dobiti i troškova dizajniranih modela utvrđena je opravdanost programa, jer je koeficijent odnosa dobiti i troškova bio pozitivan za oba modela.