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U ovom ispitivanju vršeno je uzorkovanje briseva poreklom od klinički zdravih riba koje su poticale iz različitih sredina (ribnjaci, akvarijumi, riblje pijace). Izvršena je izolacija bakterija koje su sastavni deo mikrobioma kože, škrga i creva riba i ispitivana je osetljivost ovih bakterija na određeni broj antibiotika koji se koriste u veterinarskoj i humanoj medicinskoj praksi. Precizna identifikacija ispitivanih sojeva bakterija vršena je primenom metoda PCR, sekvenciranje gena za 16S rRNK, MALDI-TOF. Primenom disk difuzionog testa i E testa ispitivano je fenotipsko ispoljavanje rezistencije na karbapeneme, ureidopeniciline sa i bez inhibitora betalaktamaza, cefalospirine III i IV generacije, aminoglikozide, tetraciklin, kolistin, flurohinolone i hloramfenikol. Prisustvo gena rezistencije, njihova lokalizacija (na hromozomu ili na mobilnim genetičkim elementima) vršena je primenom metode PCR. Kod sojeva koji su ispoljili rezistenciju na nabrojane antibiotike traženi su plazmidi i ispitivana je konjugabilnost izolovanih plazmida. Posmatrano na ukupan broj ispitanih sojeva u ovom istraživanju, bez obzira na rod i vrstu bakterija, ukupno je nađeno 55% sojeva koji su bili osetljivi na sve antibiotike, kod 22,8% sojeva nađena je rezistencija na 3 do 16 antibiotika uključujući i antibiotike koji se koriste isključivo kod ljudi (karbapenemi, ureidopenicilini, cefalosporini III i IV generacije). Ukupno 22,2% sojeva bilo je rezistentno na 1 do 2 antibiotika, mada je i među tim sojevima bilo onih koji su bili rezistentni na antibiotike registrovane samo za upotrebu kod ljudi (ceftazidim, piperacilin). Kod soja A. hydrophila izolovanom iz akvarijumske ribice gupi potvrđen je mehanizam rezistencije nalazom gena rmtB koji je bio lokalizovan na transpozonu Tn1548 smeštenom na konjugabilnom plazmidu koji je po tipu replikona bio kategorisan u grupu IncL/M. Kod sojeva Pseudomonas koji su bili rezistentni na karbapeneme, ureidopeniciline sa i bez inhibitora betalaktamaza, kao i, During this research, a series of microbiological smears was collected from clinically healthy fish found in different environments (aquaculture ponds, aquariums, and fish markets) has been done. Bacteria which belong to skin microbiome, gills, and fish intestines have been isolated, and their sensitivity to several antibiotics used in veterinary and human practice has been tested. Precise identification of tested strains of bacteria has been performed with PCR method, 16S rRNA gene sequencing and MALDI-TOF. Phenotypical manifestation of resistance to carbapenems, ureidopenicillins (with and without inhibitors of beta-lactamase), cephalosporins of the third and fourth generation, aminoglycosides, tetracyclines, colistin, fluoroquinolones and chloramphenicol has been tested using disc diffusion method and E test. Presence of resistant genes, their localization (on chromosome or on mobile genetic elements) has been conducted with PCR method. For strains showing resistance to the antibiotics mentioned above, plasmids have been searched and conjugation of isolated plasmids has been tested. Observing the total number of tested strains in this research, regardless of the genus and species of bacteria, 55% of examined strains were found to be sensitive to all antibiotics, and in 22.8% of strains resistance was observed to 3-16 antibiotics, including antibiotics used exclusively in human medicine (carbapenems, ureidopenicillins, cephalosporins of third and fourth generation). In additional 22.2% of strains the resistance to 1 or 2 antibiotics was recorded, including resistance to antibiotics registered for exclusive use in human medicine (ceftazidime, piperacillin). In A. hydrophila strain isolated from aquarium fish guppy that showed resistance to all 16 antibiotics, a mechanism of resistance has been confirmed by identifying gene rmtB which has been localized on transposon Tn1548 located on conjugal plasmid which belongs to group IncL/M type of replicon. In Pseudomonas st

During this research, a series of microbiological smears was collected from clinically healthy fish found in different environments (aquaculture ponds, aquariums, and fish markets) has been done. Bacteria which belong to skin microbiome, gills, and fish intestines have been isolated, and their sensitivity to several antibiotics used in veterinary and human practice has been tested. Precise identification of tested strains of bacteria has been performed with PCR method, 16S rRNA gene sequencing and MALDI-TOF. Phenotypical manifestation of resistance to carbapenems, ureidopenicillins (with and without inhibitors of beta-lactamase), cephalosporins of the third and fourth generation, aminoglycosides, tetracyclines, colistin, fluoroquinolones and chloramphenicol has been tested using disc diffusion method and E test. Presence of resistant genes, their localization (on chromosome or on mobile genetic elements) has been conducted with PCR method. For strains showing resistance to the antibiotics mentioned above, plasmids have been searched and conjugation of isolated plasmids has been tested. Observing the total number of tested strains in this research, regardless of the genus and species of bacteria, 55% of examined strains were found to be sensitive to all antibiotics, and in 22.8% of strains resistance was observed to 3-16 antibiotics, including antibiotics used exclusively in human medicine (carbapenems, ureidopenicillins, cephalosporins of third and fourth generation). In additional 22.2% of strains the resistance to 1 or 2 antibiotics was recorded, including resistance to antibiotics registered for exclusive use in human medicine (ceftazidime, piperacillin). In A. hydrophila strain isolated from aquarium fish guppy that showed resistance to all 16 antibiotics, a mechanism of resistance has been confirmed by identifying gene rmtB which has been localized on transposon Tn1548 located on conjugal plasmid which belongs to group IncL/M type of replicon. In Pseudomonas st, U ovom ispitivanju vršeno je uzorkovanje briseva poreklom od klinički zdravih riba koje su poticale iz različitih sredina (ribnjaci, akvarijumi, riblje pijace). Izvršena je izolacija bakterija koje su sastavni deo mikrobioma kože, škrga i creva riba i ispitivana je osetljivost ovih bakterija na određeni broj antibiotika koji se koriste u veterinarskoj i humanoj medicinskoj praksi. Precizna identifikacija ispitivanih sojeva bakterija vršena je primenom metoda PCR, sekvenciranje gena za 16S rRNK, MALDI-TOF. Primenom disk difuzionog testa i E testa ispitivano je fenotipsko ispoljavanje rezistencije na karbapeneme, ureidopeniciline sa i bez inhibitora betalaktamaza, cefalospirine III i IV generacije, aminoglikozide, tetraciklin, kolistin, flurohinolone i hloramfenikol. Prisustvo gena rezistencije, njihova lokalizacija (na hromozomu ili na mobilnim genetičkim elementima) vršena je primenom metode PCR. Kod sojeva koji su ispoljili rezistenciju na nabrojane antibiotike traženi su plazmidi i ispitivana je konjugabilnost izolovanih plazmida. Posmatrano na ukupan broj ispitanih sojeva u ovom istraživanju, bez obzira na rod i vrstu bakterija, ukupno je nađeno 55% sojeva koji su bili osetljivi na sve antibiotike, kod 22,8% sojeva nađena je rezistencija na 3 do 16 antibiotika uključujući i antibiotike koji se koriste isključivo kod ljudi (karbapenemi, ureidopenicilini, cefalosporini III i IV generacije). Ukupno 22,2% sojeva bilo je rezistentno na 1 do 2 antibiotika, mada je i među tim sojevima bilo onih koji su bili rezistentni na antibiotike registrovane samo za upotrebu kod ljudi (ceftazidim, piperacilin). Kod soja A. hydrophila izolovanom iz akvarijumske ribice gupi potvrđen je mehanizam rezistencije nalazom gena rmtB koji je bio lokalizovan na transpozonu Tn1548 smeštenom na konjugabilnom plazmidu koji je po tipu replikona bio kategorisan u grupu IncL/M. Kod sojeva Pseudomonas koji su bili rezistentni na karbapeneme, ureidopeniciline sa i bez inhibitora betalaktamaza, kao i

Prvi podaci o oboljenju prouzrokovanom O. rhinotracheale datiraju iz 1991. godine kada je u južnoj Africi, Jan du Preez (Žan di Prez) otkrio novo oboljenje brojlera koje se manifestovalo blagim respiratornim simptomima. Bakteriološkim ispitivanjem organa obolelih brojlera otkrivena je gram-negativna, pleomorfna, štapiasta bakterija, sporog rasta, koja se nije mogla identifikovati kao neka od poznatih bakterijskih vrsta. Današnji naziv Ornithobacterium rhinotracheale prihvaen je 1994. godine, kada je izvršena i klasifikacija uzronika. Prema podacima iz literature infekcija O. rhinotracheale dovodi do znaajnih ekonomskih gubitaka u živinarskoj proizvodnji koji su posledica poveanog mortaliteta, suprimiranog rasta, smanjene proizvodnje jaja i odbacivanja trupova na klanici. U literaturi ima veoma malo podataka o klinikoj slici kod brojlerskih roditelja, dok podaci o patomorfološkim promenama ne postoje. Prema navodima iz literature O. rhinotracheale je izolovan iz nekoliko razliitih vrsta divljih ptica, ali nema podataka o ornitobakteriozi fazana. Zbog toga su sprovedena ispitivanja imala za cilj da se utvrdi uticaj O. rhinotracheale na razvoj patomorfoloških promena u respiratornim organima i zdravstveno stanje kokoši brojlerskih roditelja i fazana iz odgoja. Za eksperiment je odabrano 22 fazana starosti 19 nedelja i 22 kokoši brojlerskih roditelja hibrida „Ross 308“ starosti 15 nedelja. Ptice su podeljene u etiri grupe koje su držane na podnom sistemu, u odvojenim prostorijama. Nakon sedmodnevne aklimatizacije, inficirana je po jedna grupa od 16 fazana i 16 brojlerskih roditelja intratrahealnom aplikacijom suspenzije 48-asovne kulture O. rhinotracheale, soj B3263/91 u sterilnom fiziološkom rastvoru, u koncentraciji od 5 X 1011 CFU/ml. Po jedna grupa od šest fazana i šest brojlerskih roditelja služile su kao kontrolne grupe kojima je intratrahealno aplikovan po 1 ml sterilnog fiziološkog rastvora. Posle inficiranja ptice su svakodnevno opservirane i merena im je teles, The first data relating to disease caused by O. rhinotracheale originate from 1991, when Jan du Preez discovered a new disease in broiler chickens in South Africa, which manifested itself with mild respiratory symptoms. Bacteriological examination of infected birds’ organs revealed a gram-negative, pleomorphic, slow growing, rod bacterium, which could not be identified as any of the known bacterial species. The present name Ornithobacterium rhinotracheale was accapted and the agent was classified in 1994. The literature states that the O. rhinotracheale infection can cause significant economic losses in the poultry industry as a consequence of increased mortality, suppressed growth, reduced egg production and increased carcase condemnation rate at processing. There are few data about clinical features in broiler breeders nor can data about the gross lesions be found in the literature. According to the literature, O. rhinotracheale has been isolated from several different wild bird species, but there are no data relating to ornithobacteriosis in pheasants. For these reasons investigations were carried out to determine the influence of O. rhinotracheale on the development of pathomorphological lesions in the respiratory organs and on the health status of broiler breeders and pheasants from the rearing stage. A total of 22 pheasants, 19 weeks of age and 22 "Ross 308" broiler breeder hens, 15 weeks of age were used in this study. The birds were divided into four groups which were housed in a floor system, in separate rooms. After seven days of acclimatization, one group of 16 pheasants and 16 broiler breeder hens were infected intratracheally with a suspension of 48-hour culture of O. rhinotracheale strain B3263/91 in sterile saline in concentration of 5 X 1011 CFU/ml. Six pheasants and six broiler breeders served as control groups and were inoculated intratracheally with 1 ml of sterile saline per bird. After infection, the birds were observed daily and weighed at the

Cilj ovog ispitivanja je bio da se ustanovi prisustvo S. suis na nekim farmama iklanicama svinja u Republici Srbiji, definišu biohemijske osobine izolovanih sojeva, utvrdinjihova serotipska pripadnost i ispita njihova osetljivost na antibiotike. Poznato je da se S.suis nalazi kao normalan stanovnik respiratornog sistema svinja, najčešće tonzila i nosnihšupljina, a često se može izolovati i iz genitalnog i gastrointestinalnog sistema. Kliničkizdrave svinje predstavljaju glavni rezervoar uzročnika i najznačajniju kariku uepidemiologiji infekcije kod ljudi. Iako se smatra uglavnom patogenom svinja, S. suis je dosada izolovan kao uzročnik infekcija kod različitih drugih vrsta sisara (goveda, mačaka,ovaca) i ptica. Infekcije kod ljudi izazvane S. suis smatrane su sporadičnim i bile suuglavnom vezane za ljude koji dolaze u kontakt sa svinjama i njihovim proizvodima.Međutim, 2005. godine u Kini je izbila epidemija izazvana S. suis kada je obolelo više od 200ljudi, a kod skoro 20% obolelih bolest se završila fatalno. Ova epidemija je potpunopromenila shvatanje o opasnosti ovog patogena za zdravlje ljudi. Veterinarske dijagnostičkelaboratorije obično lako identifikuju S. suis upotrebom malog broja biohemijskih testova,naročito ukoliko su ispitivani uzorci poreklom od obolelih svinja. Međutim, smatra se da nekisojevi (i/ili serotipovi) S. suis mogu biti pogrešno identifikovani komercijalnim kitovima, zbogčega serotipizacija, koja se inače koristi i u rutinskoj dijagnostičkoj proceduri predstavljajednu od sigurnih dijagnostičkih metoda Osetljivost na antibiotike sojeva S. suis u zemljamau kojima je ispitivana uglavnom je zavisila od antibiotika koji su korišćeni u lečenju svinja.Za ovo ispitivanje korišćeni su brisevi nazofaringealnih tonzila prasadi i svinja, nosnihšupljina prasadi, ali i brisevi organa uginule prasadi, brisevi sa trupova zaklanih svinja ibrisevi sa noža za evisceraciju organa na klanici. Svi uzorci su transportovani u tripton sojabujonu u roku od 2 časa od u, The aim of this study was to establish the presence of S. suis on some pig farms in theRepublic of Serbia, then, to define their biochemical profiles, determine their serotypeaffiliation and test antimicrobial susceptibility of isolated serotypes. It is known that S. suis isa normal inhabitant of the respiratory system of pigs, mostly of the tonsils and nasal cavities,and can often be isolated from the genital and gastrointestinal system. Clinically healthy pigsare a major reservoir of pathogens and the most important link in the epidemiology ofinfection in humans. Although generally considered a pathogen for pigs, S. suis has beenisolated as the cause of infection in a variety of other mammal species (cattle, cats, sheep)and birds. Infections in humans caused by S. suis were considered sporadic and were mostlyrelated to people who come in contact with pigs and their products. However, in year 2005the outbreak in China caused by S. suis affected more than 200 people, with almost 20% ofpatients with the disease ending fatally. This epidemic has completely changed the perceptionof the danger of this pathogen to human health. Veterinary diagnostic laboratories usuallyeasily identify S. suis using a small number of biochemical tests, especially if the samplesoriginate from diseased pigs. However, it is considered that some S. suis strains (and/orserotypes) may be incorrectly identified by commercial kits, thus making serotyping(normally used in routine diagnostic procedures), one of the safest diagnostic procedures.For this test we used nasopharyngeal swabs of pigs and pig tonsils, nasal cavities of piglets,and swabs of dead pigs, swabs from the carcasses of slaughtered pigs and swabs from theevisceration knife after slaughter. All samples were transported in tryptone soy broth within 2hours of sampling. For isolation were used solid and liquid media (Colombia agar with 5%sheep blood added, brain heart infusion with 6.5% NaCl added, tryptone soy broth, esculinbroth, and

For the purpose of detecting methicillin (oxacillin) resistance in staphylococcal strains, in a number of microbiological laboratories only disc diffusion method with cefoxitin and/or oxacillin discs is used. Besides this method, it is desirable to determine MIC values for cefoxitin and/or oxacillin. After examination by disc diffusion and dilution methods, latex agglutination is used for the detection of PBP2a and PCR is used for the detection of mecA gene. Use of PCR is not possible in a large number of diagnostic laboratories and as method of choice, latex agglutination test for rapid detection of PBP2a is recommended. In this investigation, as confirmatory methods, latex agglutination and PCR were used for strains that were resistant to oxacillin and/or cefoxitin by disc diffusion and broth microdilution methods. In total, 14 strains of coagulase-negative staphylococci originating from clinical specimens of cats, dogs and chicken were examined. Among isolated strains, it was established that the dominating species was Staphylococcus haemolyticus with 11 isolated strains. Other isolated species were Staphylococcus epidermidis, Staphylococcus capitis and Staphylococcus vitulinus, each with one isolated strain. For all 14 strains, oxacillin MIC values ranged from 0.5 μg/mL to >64 μg/mL and cefoxitin MIC values ranged from 1 μg/mL to >256 μg/mL. Positive agglutination reaction by latex agglutination test was recorded in 13 out of 14 strains. The PCR assay for mecA gene was positive in 12 investigated strains., Radi otkrivanja meticilin (oksacilin)-rezistentnih sojeva stafilokoka u većini mikrobioloških laboratorija koristi se najčešće samo disk difuziona metoda uz primenu antibiogram diskova oksacilina i cefoksitina. Pored navedene metode, poželjno je da se utvrde i vrednosti MIC oksacilina i/ili cefoksitina primenom dilucione metode u bujonu ili agaru. Nakon ispitivanja pomenutim metodama za definitivno utvrđivanje pripadnosti soja grupi meticilin rezistentnih stafilokoka, koriste se metoda lateks aglutinacije za utvrđivanje prisustva PBP2a i PCR metoda za detekciju mecA gena. Kako primena metode PCR nije moguća u većini laboratorija koje se bave rutinskom dijagnostikom, kao metoda izbora preporučuje se lateks aglutinacioni test za brzo otkrivanje PBP2a. U ovom ispitivanju primenjeni su lateks aglutinacioni test i PCR metod, kao potvrdne metode za ispitivanje sojeva stafilokoka koji su bili rezistentni na oksacilin i/ili cefoksitin na osnovu rezultata dobijenih disk difuzionom metodom i mikrodilucionom metodom u bujonu. Ukupno je ispitano 14 sojeva koagulaza - negativnih stafilokoka izolovanih iz kliničkih uzoraka poreklom od pasa, mačaka i kokoši. Među izolovanim sojevima, ustanovljeno je da dominira vrsta Staphylococcus haemolyticus sa 11 izolata, a po jedan soj je izolovan od vrsta Staphylococcus epidermidis, Staphylococcus capitis i Staphylococcus vitulinus . Vrednosti MIC oksacilina za 14 sojeva iznosile su od 0.5 mg/mL do >64 mg/mL, a vrednosti MIC cefoksitina iznosile su od 1 mg/mL do >256 mg/mL. Kod 13 od 14 sojeva utvrđena je pozitivna reakcija aglutinacije primenom lateks aglutinacionog testa. Kod 12 ispitivanih sojeva je utvrđeno prisustvo mecA gena PCR metodom.

Four pooled samples of whole poultry carcasses with their internal organs were used to determine the presence of Newcastle disease (ND) virus. Samples were collected from one epizootiological area in the Republic of Serbia during January 2007. Newcastle disease virus strains were isolated from four samples. The identification of isolated strains was done by using the hemagglutination and hemagglutination-inhibition tests. The nucleic acid of the ND virus was identified in all the four samples It was confirmed that all the isolated strains were velogenic strains. Analysis of the nucleotide sequences of the gene encoding the F cleavage site of the fusion F protein showed the presence of motifs 112RRQKRFIG119, characteristic for the velogenic strains of the ND virus. Phylogenetic analysis of the F gene sequences revealed that all isolated strains of the virus belong to class II and genotype VIId.

The objective of this work was to identify isolated Staphylococcus species and to investigate their sensitivity to some antibacterial drugs. The material used for these investigations were Staphylococcus isolates originating from milk samples. A total of 25 strains of Staphylococcus isolates were examined, including 24 from milk samples from cows with mastitis, and one strain was isolated from a milk sample from a cow following treatment for mastitis. For primary identification, catalase and oxidase tests were used, as well as the free coagulase test. Following the preliminary tests, the isolated strains were identified using commercial systems ID32 STAPH (bioMérieux, France) and the BBL Crystal Gram-Positive ID Kit (Becton Dickinson, USA) according to the enclosed instructions. The Staphylococcus isolates were examined for sensitivity to the following: oxacillin, penicillin, cefoxitin, gentamicin, erythromycin, chloramphenicol, tetracycline, ciprofloxacin, sulfametoxazol/trimetoprim, and vacomycin using the disk diffusion method and the broth microdilution method as recommended by the Clinical and Laboratory Strandards Institute - CLSI(2003), and the results were interpreted according to CLSI recommendations from 2008 and 2010. Antibiogram disks manufactured by Becton Dickinson (USA) were used, and the broth microdilution method was applied using pure antibiotic substances from different manufacturers: erythromycin, chloramphenicol, cefoxitin, gentamicin, oxacillin, tetracycline (Sigma Aldrich, USA), sulfametoxazol (Fluka, USA), penicillin (Calbiochem, Germany), vancomycin (Abbott laboratories, USA), ciprofloxacin and trimetoprim (Zdravlje A.D., Serbia). All 25 strains were catalase positive and oxidase negative. Of the 25 strains, 19 were coagulase positive and 6 were coagulase negative.With the implementation of the disk diffusion method on 19 strains of S. aureus, 17 were established to be resistant to penicillin (89.5%), and 2 strains to gentamicin (10.5%). The, Cilj ovog rada je bila identifikacija izolovanih vrsta stafilokoka i ispitivanje njihove osetljivosti na neke antibakterijske lekove. Kao materijal u ovom ispitivanju korišćeni su izolati stafilokoka poreklom iz uzoraka mleka. Ukupno je ispitano 25 sojeva izolovanih stafilokoka od kojih su 24 poticala iz uzoraka mleka krava sa mastitisom, a jedan soj je izolovan iz uzorka mleka krave nakon lečenja mastitisa. U primarnoj identifikaciji su korišćeni katalaza i oksidaza testovi, kao i test prisustva slobodne koagulaze. Nakon izvođenja preliminarnih testova, vršena je identifikacija izolovanih sojeva, primenom komercijalnih sistema ID32 STAPH (bioMérieux, Francuska) i BBL Crystal Gram-Positive ID Kit (Becton Dickinson, SAD) prema uputstvima proizvođača. Osetljivost izolovanih sojeva stafilokoka ispitivana je na: oksacilin, penicilin, cefoksitin, gentamicin, eritromicin, hloramfenikol, tetraciklin, ciprofloksacin, sulfametoksazol/trimetoprim i vankomicin primenom disk difuzione metode i mikrodilucione metode u bujonu prema preporukama Instituta za kliničke i laboratorijske standarde (Clinical and Laboratory Strandards Institute - CLSI (2003), a tumačenje rezultata je vršeno prema preporukama CLSI iz 2008. i 2010. godine. Korišćeni su antibiogram diskovi proizvođača Becton Dickinson (SAD), a za mikrodilucionu metodu u bujonu korišćene su čiste supstance antibakterijskih lekova različitih proizvođača: eritromicin, hloramfenikol, cefoksitin, gentamicin, oksacilin, tetraciklin (Sigma Aldrich, SAD), sulfametoksazol (Fluka, SAD), penicilin (Calbiochem, Nemačka), vankomicin (Abbott laboratories, SAD), ciprofloksacin i trimetoprim (Zdravlje A.D., Srbija). Svih 25 sojeva je bilo katalaza pozitivno i oksidaza negativno. Od 25 sojeva, 19 je bilo koagulaza-pozitivno, a 6 koagulaza-negativno. Primenom disk difuzione metode od 19 sojeva S. aureus kod 17 je utvrđena rezistencija na penicilin (89,5%), a kod 2 soja na gentamicin (10,5%). Od 3 soja S. xylosus, kod jednog je primenom disk

Streptococcus suis is a normal inhabitant of the respiratory system of pigs. Clinically healthy pigs are a major reservoir of the pathogen, but, as well, the most important link in the epidemiology of infection in humans. Hence, the aim of this study was to establish the presence of S. suis on some pig farms in Serbia and to determine their serotype affiliation. For this research, we tested 226 samples: nasopharyngeal tonsil swabs, nasal cavities swabs, parts of or all nasopharyngeal tonsils, swabs from dead animals, swabs from pig carcasses, and swabs from the butchers knives. Identification of S. suis strains was performed with antisera specific for capsular antigens. S. suis serotype 2 was established in 67.7% cases, and serotypes 7, 9 and 1 were isolated in a much smaller percentage (17.6, 8.8 and 5.9%, respectively). Serotype 2 is the only serotype isolated from all the collected samples. Serotype 9 was isolated from healthy pig tonsil swab and from 2 nasal swabs, and was not found in the samples taken from diseased pigs. The serotype 1 was isolated only from nasopharyngeal tonsil swabs of piglets, while serotype 7 was isolated from nasopharyngeal tonsil swabs, nasopharyngeal tonsil sections, but also from nasal swabs of healthy pigs. It can be concluded that the infections caused by different serotypes of S. suis are present in the herd of pigs in Serbia; hence, it is very important to identify what serotypes of S. suis are involved in pathogenesis of infections in pigs.

Methicillin-resistant Staphylococcus aureus is a highly important human pathogen that is also a significant concern in veterinary medicine. Despite the high prevalence of colonization, clinical infections with methicillin-resistant Staphylococcus aureus appear to be rare in pigs. Methicillin-resistant Staphylococcus aureus was isolated from a sow with endometritis and her five piglets with dermatitis originating from a Serbian farm. Identification of the strains was done by automated system and confirmed by polymerase chain reaction for mecA and nuc genes. Detection of Staphylococcal Cassette Chromosome mec type was performed by multiplex polymerase chain reaction. Antimicrobial susceptibility testing on erythromycin, clindamycin, gentamicin, kanamycin, tobramycin, ciprofloxacin, tetracycline, trimethoprim/sulfamethoxazole and vancomycin was done by disc diffusion method. Six isolated strains from the infected sow and her piglets showed resistance only to tetracycline beside resistance to all beta-lactam antibiotics. In the tested methicillin-resistant Staphylococcus aureus isolates, Staphylococcal Cassette Chromosome mec type V was present. To our knowledge, this finding is the first documented detection of methicillin-resistant Staphylococcus aureus from pigs' clinical samples in Serbia. The results of our study indicate the emergence of methicillin-resistant Staphylococcus aureus in a pig farm in Serbia highlighting the threat of this antibiotic-resistant microorganism as a pathogen causing both animal and human infections.

Cilj ovog rada je bilo ispitivanje prisustva i zastupljenosti streptokoka, enterokoka i streptokokama sličnih bakterija u različitim materijalima poreklom od zdravih, zaklanih i uginulih svinja različitih kategorija sa nekoliko farmi i klanica u Republici Srbiji. Ukupno je ispitano 226 uzoraka briseva tonzila i nosa od klinički zdrave prasadi u tovu, briseva tonzila prasadi starosti 5-20 dana, delova nazofaringealnih tonzila tovljenika zaklanih na klanici, delova nazofaringealnih tonzila prasadi zaklanih na klanici, briseva sa trupova zaklanih svinja na klanici, briseva sa no`eva za evisceraciju organa na klanici, kao i briseva pluća, trbušne duplje i organa naglo uginule prasadi. Ispitivanje prisustva navedenih vrsta bakterija vršeno je primenom standardnih mikrobioloških metoda. Za identifikaciju izolovanih vrsta bakterija korišćeni su komercijalni biohemijski testovi, a za utvrđivanje serotipske pripadnosti izolovanih sojeva Streptococcus suis specifični serumi za kapsularne antigene. Ustanovljeno je da najveći broj izolovanih sojeva pripada rodu Streptococcus, a u daleko manjem broju izolovane bakterije su pripadale rodovima: Enterococcus (6) (10,4%), Aerococcus (3) (6,2%), Lactococcus (2) (4,2%) i Globicatella (2) (4,2%). Od navede nog broja Streptococcus vrsta najveći broj je pripadao vrsti Streptococcus suis (23) (64%). U brisevima nazofaringealnih tonzila utvrđeno je prisustvo i ostalih á hemolitičkih streptokoka: Streptococcus sanguinis (13,8%), Streptococcus salivarius (5,6%), Streptococcus mitis (5,6%), Streptococcus parasanguinis (2,7%) i Streptococcus oralis (2,7%). Takođe, u manjem procentu izolovana je i vrsta Streptococcus bovis (5,6%). Najveći procenat izolovanih bakterija iz roda Enterococcus pripadao je vrsti Enterococcus faecalis (80%), dok je vrsti Enterococcus faecium pripadao znatno manji procenat izolata (20%). Od ostalih takozvanih streptokokama sličnih vrsta bakterija izolovane su: Aerococcus viridans, Globicatella sanguinis i Lactococcus l, The aim of this investigation was to establish the presence and prevalence of streptococci, enterococci and streptococci-like bacteria in various materials originating from healthy, slaughtered and dead pigs belonging to different categories from several farms and slaughterhouses in the Republic of Serbia. The total number of investigated samples comprised 226 swabs of tonsils and noses from clinically healthy breeders, swabs of tonsils from piglets 5-20 days old, parts of nasopharyngeal tonsils from breeders slaughtered in a slaughterhouse, parts of nasopharyngeal tonsils from piglets slaughtered in a slaughterhouse, swabs of slaughtered pig carcasses from a slaughterhouse, swabs from knives for evisceration in a slaughterhouse, as well as swabs of lungs, abdominal cavity and organs from piglets which died suddenly. The standard microbiological methods were used for investigations of the presence of the listed microorganisms. Commercial biochemical tests were used for the identification of the isolated bacteria and specific sera for capsular antigenes were used for serological determination of the isolated S. suis strains. It was established that the great majority of the isolated strains belonged to the genus Streptococcus (36) (75%), and the minority of the strains belonged to the following genera: Enterococcus (6) (10.4%), Aerococcus (3) (6.2%), Lactococcus (2) (4.2%) and Globicatella (2) (4.2%). The great majority of Streptococcus species belonged to S. suis. The presence of other á haemolytic streptococci was established in the swabs of nasopharyngeal tonsils: Streptococcus sanguinis (13.8%), Streptococcus salivarius (5.6%), Streptococcus mitis (5.6%), Streptococcus parasanguinis (2.7%) and Streptococcus oralis (2.7%). Also, S. bovis was isolated in a smaller percentage (5.6%). The greatest number of isolated bacteria from the genus Enterococcus belonged to Enterococcus faecalis (80%), while the minority of isolated strains belonged to Enterococcus faecium (20

Listeria monocytogenes is a bacterium that is pathogenic for man and for most animal species. Listeriosis is a generalized infection that starts after ingestion of the causative agent L. monocytogenes (Finlay, 2001). Food that is not properly thermically processed, long storage of such food, food that is produced in inadequate hygienic food plants, as well as cooked-cooled ready to eat food (RTE) is the ideal medium for listeria multiplication. High risk food originated from fish, fish products and sea products comprised of: molluscs (fresh of frozen shellfish, crustaceans shelled or not), fresh fish (ready to eat without cooking), fish products packed with brine (NaCl lt 6%), (salted, marinated, fermented, cold smoked and fish in brine), thermically treated fish and crustacean products (pasteurization, cooking, hot smoking, including pre-cooking and panning). In this research, fish samples, fish products and sea products from Serbian markets were examinated such as: fresh fish (cooled), frozen food (fish and sea products - cuttlefish, squid, octopus, shellfish, crustaceans and shrimps), panned products; smoked fish, salted fish, thermally treated fish and fish products, semi-canned fish and canned fish. Microbiological testing has been performed according to internationally prescribed standards ISO 11290-1 (1996) 'Microbiology of food and animal feed stuffs - Horizontal method for the detection and enumeration of Listeria monocytogenes'. There were 470 samples from fish products and sea products. Presence of Listeria spp was registered in 58 samples (12.34%). Listeria monocytogenes was found in 1.92% samples (9 isolates), which makes 15.52% of the total Listeria that were isolated from the tested food. Other isolated Listeria belong to the following species: L. innocua (8.51%), L.welshimeri (1.28%), L.welshimeri/innocua (0.21%), L. grayi (0.21%) and L. seeligeri (0.21%). Exceptional viability of Listeria monocytogenes in the food tested was documented, also (freezi, Listeria monocytogenes je bakterija patogena za ljude i veliki broj životinjskih vrsta. Listerioza predstavlja generalizovanu infekciju koja nastaje nakon oralne ingestije uzročnog agensa L. monocytogenes (Finlay, 2001). Navedena bakterija se najbolje razvija u namirnicama koje nisu adekvatno termički obrađene; koje su dugo skladištene; koje su proizvedene na mestu na kojem principi higijenskog rukovanja hranom nisu sprovođeni; kuvano-hladnoj hrani spremnoj za jelo (ready to eat food - RTE) U visoko rizične namirnice počev od vrsta ribe, proizvoda od ribe i plodova mora ubrajaju se: mekušci (sveže i zamrznute školjke, rakovi u ljušturama, ili očišćeni), sveža riba (koja se konzumira bez termičke obrade), riblji proizvodi u vodenom rastvoru soli (NaCl%), (soljena, marinirana, fermentisana, hladno dimljena i riba u sopstvenom soku), srednje termički obrađeni riblji proizvodi i rakovi (pasterizacija, kuvanje, toplo dimljenje, uključujući i pred-kuvanje i panirani proizvodi). Kao materijal za ispitivanje korišćeni su uzorci riba, proizvoda od riba i morski plodovi: sveža riba (u rashlađenom stanju), zamrznute namirnice (riba i morski plodovi - lignje, sipe, hobotnice, školjke, rakovi i škampi), panirani proizvodi, dimljena riba, usoljena riba, termički obrađena riba i proizvodi od ribe, polukonzerve i konzerve od ribe. Mikrobiološko ispitivanje je rađeno prema propisanoj metodi po Međunarodnom standardu ISO 11290-1 (1996) 'Microbiology of food and animal feeding stuffs - Horizontal method for the detection and enumeration of Listeria monocytogenes'. Ukupno je ispitano 470 uzoraka ribe, proizvoda od ribe i morskih plodova. Prisustvo Listeria vrsta utvrđeno je u 58 uzoraka (12,34%). Listeria monocytogenes je utvrđena kod 1,92% pregledanih uzoraka (9 izolata) i čini 15,52% od svih vrsta iz roda Listeria koje su izolovane iz navedenih namirnica. Ostale vrste listerija koje su izolovane pripadale su: L. innocua (8,51%), L. welshimeri (1,28%), L. welshimeri/innocua (0,21%), L

Resistance to antibiotics is not a modern phenomenon. On the contrary, penicillin resistance in some bacterial strains developed quickly after its introduction into daily practice. At the same time some bacterial strains developed resistance to almost all known antibiotics, vancomycin included. Vancomycin was for a long time the only efficient antibiotic against staphylococcal infections. It is of special concern the fact that antibiotics are in everyday exploitation in agriculture and veterinary clinical practice which use them not only as a mean of therapeutic treatment, but as an additive in animal feedstuffs in order to promote growth and prevent bacterial infections. The same antibiotics are used in human medicine, which is a persistent problem. In such a way it is possible to develop resistance which can be transferred to human pathogenic bacteria via mobile genetic elements. The incidence of resistant bacterial strains increases year after year not only on a local level, but on a global scale, as well. Monitoring of the use of antibiotics and chemotherapeutics in the Republic of Serbia is not established as such, our intention was to study a number of bacteria isolated from cattle, pigs, poultry, dogs and cats. At this time we are presenting the results for pathogenic strains of E. coli in order to determine the use of antibiotics and chemotherapeutics of the old and new generations in domestic animals. E. coli sensitivity was investigated with the disc diffusion test for: ampicillin, amoxicillin with clavulanic acid, tetracycline, chloramphenicol, gentamicin, and ceftriaxon, sulphamethoxasole with trimethoprim, ciprofloxacin and florfenicol. E. coli strains resistant to three or more antibiotics were tested by means of agar dilution method for ciprofloxacin, tetracycline, chloramphenicol, gentamicin and amoxicillin with clavulanic acid by determination of the minimal inhibitory concentration (MIC). The tested E. coli strains resulted resistant to all antibio, Rezistencija na antibiotike nije skorašnji fenomen, naprotiv, rezistencija na penicilin kod nekih sojeva bakterija javila se vrlo brzo posle njegove primene u praksi. Takođe, neki sojevi bakterija postali su rezistentni na praktično sve antibiotike u kliničkoj upotrebi, pa čak i na vankomicin koji je duže vreme bio jedini efikasan antibiotik u lečenju stafilokoknih infekcija. Poseban razlog za zabrinutost predstavlja upotreba antibiotika u poljoprivredi i veterinarskoj kliničkoj praksi jer se osim primene u lečenju životinja, iako su zabranjeni, koriste kao promoteri rasta i u prevenciji bakterijskih infekcija, a isti antibiotici (ili sa istim načinom delovanja na bakterije) primenjuju se i u humanoj medicini. Na taj način je moguće da se neodgovornom upotrebom antibiotika razvije rezistencija koja može znatno brže da se prenese na patogene bakterije ljudi preko mobilnih genetičkih elemenata. Pojava rezistentnih sojeva bakterija se povećava iz godine u godinu, kako na lokalnom, tako i na globalnom nivou. Kako monitoring korišćenja antibiotika i hemioterapeutika u veterinarskoj medicini u Republici Srbiji nije uspostavljen, a problem rezistencije bakterija na antibakterijske lekove je prisutan, ovim ispitivanjem želeli smo da obuhvatimo više vrsta bakterija izolovanih od goveda, svinja, živine, pasa i mačaka. Rezultate dobijene u ovom ispitivanju iznosimo samo za patogene sojeve E. coli, kako bi na indirektan način ustanovili primenu antibiotika i hemioterapeutika starije i novije generacije kod pomenutih životinja. Osetljivost navedenih sojeva E. coli ispitivana je na: ampicilin, amoksicilin sa klavulanskom kiselinom, tetraciklin, hloramfenikol, gentamicin, ceftriakson, sulfometoksazol sa trimetoprimom, ciprofloksacin i florfenikol, primenom disk difuzione metode. Kod sojeva E. coli kod kojih je primenom disk-difuzione metode ustanovljena rezistencija na tri i više antibiotika, odnosno multi rezistencija dalje ispitivanje je vršeno i dilucionom metodom u agaru na: c

Avian paramyxoviruses type 1 or Newcastle disease viruses (NDV) are frequently recovered from wild birds and such isolates are most frequently of low virulence. Velogenic NDV are usually recovered from poultry and only occasionally from wild birds. Five NDV isolates were obtained from carcasses of four wild bird species during 2007 in Serbia: Mallard (Anas platyrhynchos), Eurasian Sparrowhawk (Accipiter nisus), feral Rock Pigeon (Columba livia), and Eurasian Collared Dove (Streptopelia decaocto). All the isolates have a typical fusion protein cleavage site motif of velogenic viruses (R-112-R-Q-K-R-F-117). The highest homology (99%) for the nucleotide sequences spanning the M and F gene of the studied isolates was with the genotype VII NDV isolate Muscovy duck/China(Fujian)/FP1/02. Phylogenetic analysis based on a partial F gene sequence showed that the isolates from wild birds cluster together with concurrent isolates from poultry in Serbia within the subgenotype VIId, which is the predominant pathogen involved currently in Newcastle disease outbreaks in poultry worldwide. It is unlikely that the wild birds played an important role in primary introduction or consequent spread of the velogenic NDV to domestic poultry in Serbia, and they probably contracted the virus from locally infected poultry.

Among different pathogens, enterotoxic E. coli (ETEC) has been for many years an important etiological agent in the occurrence of digestive system disease of newborn animals. In counties with developed pig breeding (farming), including our country, diarrhea in the neonatal period, caused by ETEC strains is one of the most present and economically most significant diseases. The aim of this investigation wais to determine the prevalence of ETEC strains in piglets (weaning pigs), originated from 5 (five) pig farms in the Republic of Serbia, as well as their serological typization based on characteristics of somatic O antigens, presence of fimbrial antigensadhesins and hemolytic activity. The material for this investigation was targeted and sampled from piglets that have shown clinical signs of neonatal diarrhea or pathoanatomical changes characteristic for enteritis caused by ETEC strains. The total number of isolated ETEC strains were 148, of which 91 (61.48 %) were determined on the basis of somatic O antigen characteristics. The largest number of strains, 42 (46.15 %) belonged to serotype O149. Serological types O8 and O147 were represented, each with 15 strains (16.48 %). In 13 (14.28 %) strains the somatic antigen which belonged to serotype O138 was determined and in 6 (6.59 %) strains the antigen belonged to serotype O157. No strain agglutinated with hyperimmune O139 serotype serum. The presence of fimbrial adhesins was determined in 47 (51.64%) strains and of that number the F4 type of fimbrial adhesins was detected in 38 (80.85 %) strains. The presence of F5 adhesins was determined in 4, and F6 in 3. In 2 strains, the paralell presence of two adhesin types, F4 and F6 was detected. The greatest number of strains 30 (71.42 %) with adhesin F4 belonged to O149 serotype, a considerably smaller number, 4 (26.66%) to serotype O8, 2 strains to serotype O157 and to each serotype O147 and O138 1 strain. The fimbrial adhesin of F5 type was detected in 3 strains which belo, Među različitim patogenima, enterotoksična E.coli (ETEC) je već dugo značajan etiološki agens u nastanku oboljenja digestivnog sistema novorođenih životinja. U zemljama sa razvijenom svinjarskom proizvodnjom, uključujući i našu zemlju, dijareja u neonatalnom periodu, uzrokovana sojevima ETEC je jedna od najprisutnijih i ekonomski najznačajnijih bolesti. Cilj istraživanja je bio da se utvrdi prevalencija sojeva ETEC kod prasadi, poreklom sa 5 farmi svinja u Srbiji, kao i njihova serološka tipizacija na osnovu karakteristika somatskog O antigena, prisustva fimbrijalnih antigena - adhezina i hemolitičke aktivnosti. Materijal za istraživanja ciljano je uzorkovan od prasadi koja su pokazivala kliničke znake neonatalne dijareje ili patoanatomske promene karakteristične za enteritise izazvane sojevima ETEC. Ukupno je izolovano 148 sojeva ETEC od čega je 91 soj (61,48%) bio tipiziran na osnovu karakteristika somatskog O antigena. Najveći broj sojeva, 42 (46,15%) pripadao je serotipu O149. Serološke grupe O8 i O147 su bile zastupljene sa po 15 (16,48%) sojeva. Kod 13 (14,28 %) sojeva, utvrđen je somatski antigen serogrupe O138, a kod 6 (6,59%) sojeva antigen serogrupe O157. Nijedan soj nije aglutinirao sa hiperimunim serumom serogrupe O139. Prisustvo fimbrijalnih adhezina ustanovljeno je kod 47 (51,64 %) sojeva, a od tog broja F4 tip fimbrijalnih adhezina detektovan je kod 38 (80,85 %) sojeva. Prisustvo F5 adhezina ustanovljeno je kod 4, F6 kod 3, a kod 2 soja detektovano je istovremeno prisustvo dva tipa adhezina, F4 i F6. Najveći broj sojeva, 30 (71,42 %) sa F4 adhezinom pripadao je serogrupi O149, znatno manji broj, 4 (26,66 %) serogrupi O8, 2 soja serogrupi O157 i po 1 soj serogrupama O147 i O138. Fimbrijalni adhezin tipa F5 otkriven je kod 3 soja serogrupe O8 i kod 1 soja iz serogrupe O149. Sva 3 soja sa F6 adhezinom pripadala su serogrupi O8. Od 2 soja sa istovremenim prisustvom adhezina F4 i F6, jedan je pripadao serogrupi O8, a drugi serogrupi O138. Hemolitička aktiv

Examinations were performed on 65 samples of nasal smeas taken from calves and young cows with clinical symptoms of respiratory infection to determine the presence of the bovine herpes virus 1 using parallel implementation of molecular and standard methods of virological diagnostics. The appearance of a cytopathogenic effect (CPE) was not established in inoculated cell lines 24h, 48h and 72h following inoculation, or after two successive passages of the examined material sample through these cell lines. The application of polymerize chain reaction (PCR) using a primer for glucoprotein B and thymidine - kinasis, established the presence of bovine herpes virus 1 nucleic acid in one sample of a bovine nasal smear, while the presence of this virus was established in three samples in an examination of the nasal smear samples using the Real-Time PCR method., Ukupno je ispitivano 65 uzoraka nosnih briseva prikupljenih od teladi i junadi sa kliničkim simptomima respiratorne infekcije na prisustvo goveđeg herpesvirusa 1 uporednom primenom molekularnih i standardnih metoda virusološke dijagnostike. Kod inokulisanih ćelijskih linija nije ustanovljena pojava citopatogenog efekta (CPE -) posle 24h, 48h i 72h od inokulacije ni posle dve uzastopne pasaže uzoraka ispitivanog materijala kroz navedene ćelijske linije. Primenom lančane reakcije polimeraze (PCR) uz korišćenje prajmera za glikoprotein B i timidin-kinazu, utvrđeno je prisustvo nukleinske kiseline goveđeg herpesvirusa 1 u jednom uzorku nosnog brisa, dok je ispitivanjem navedenih uzoraka nosnih briseva goveda metodom Real-Time PCR prisustvo pomenutog virusa ustanovljeno kod tri uzorka.

Među različitim patogenima, enterotoksična E.coli (ETEC) je već dugo značajan etiološki agens u nastanku oboljenja digestivnog sistema novorođenih životinja. U zemljama sa razvijenom svinjarskom proizvodnjom, uključujući i našu zemlju, dijareja u neonatalnom periodu, uzrokovana sojevima ETEC je jedna od najprisutnijih i ekonomski najznačajnijih bolesti. Cilj istraživanja je bio da se utvrdi prevalencija sojeva ETEC kod prasadi, poreklom sa 5 farmi svinja u Srbiji, kao i njihova serološka tipizacija na osnovu karakteristika somatskog O antigena, prisustva fimbrijalnih antigena - adhezina i hemolitičke aktivnosti. Materijal za istraživanja ciljano je uzorkovan od prasadi koja su pokazivala kliničke znake neonatalne dijareje ili patoanatomske promene karakteristične za enteritise izazvane sojevima ETEC. Ukupno je izolovano 148 sojeva ETEC od čega je 91 soj (61,48%) bio tipiziran na osnovu karakteristika somatskog O antigena. Najveći broj sojeva, 42 (46,15%) pripadao je serotipu O149. Serološke grupe O8 i O147 su bile zastupljene sa po 15 (16,48%) sojeva. Kod 13 (14,28 %) sojeva, utvrđen je somatski antigen serogrupe O138, a kod 6 (6,59%) sojeva antigen serogrupe O157. Nijedan soj nije aglutinirao sa hiperimunim serumom serogrupe O139. Prisustvo fimbrijalnih adhezina ustanovljeno je kod 47 (51,64 %) sojeva, a od tog broja F4 tip fimbrijalnih adhezina detektovan je kod 38 (80,85 %) sojeva. Prisustvo F5 adhezina ustanovljeno je kod 4, F6 kod 3, a kod 2 soja detektovano je istovremeno prisustvo dva tipa adhezina, F4 i F6. Najveći broj sojeva, 30 (71,42 %) sa F4 adhezinom pripadao je serogrupi O149, znatno manji broj, 4 (26,66 %) serogrupi O8, 2 soja serogrupi O157 i po 1 soj serogrupama O147 i O138. Fimbrijalni adhezin tipa F5 otkriven je kod 3 soja serogrupe O8 i kod 1 soja iz serogrupe O149. Sva 3 soja sa F6 adhezinom pripadala su serogrupi O8. Od 2 soja sa istovremenim prisustvom adhezina F4 i F6, jedan je pripadao serogrupi O8, a drugi serogrupi O138. Hemolitička aktiv, Among different pathogens, enterotoxic E. coli (ETEC) has been for many years an important etiological agent in the occurrence of digestive system disease of newborn animals. In counties with developed pig breeding (farming), including our country, diarrhea in the neonatal period, caused by ETEC strains is one of the most present and economically most significant diseases. The aim of this investigation wais to determine the prevalence of ETEC strains in piglets (weaning pigs), originated from 5 (five) pig farms in the Republic of Serbia, as well as their serological typization based on characteristics of somatic O antigens, presence of fimbrial antigensadhesins and hemolytic activity. The material for this investigation was targeted and sampled from piglets that have shown clinical signs of neonatal diarrhea or pathoanatomical changes characteristic for enteritis caused by ETEC strains. The total number of isolated ETEC strains were 148, of which 91 (61.48 %) were determined on the basis of somatic O antigen characteristics. The largest number of strains, 42 (46.15 %) belonged to serotype O149. Serological types O8 and O147 were represented, each with 15 strains (16.48 %). In 13 (14.28 %) strains the somatic antigen which belonged to serotype O138 was determined and in 6 (6.59 %) strains the antigen belonged to serotype O157. No strain agglutinated with hyperimmune O139 serotype serum. The presence of fimbrial adhesins was determined in 47 (51.64%) strains and of that number the F4 type of fimbrial adhesins was detected in 38 (80.85 %) strains. The presence of F5 adhesins was determined in 4, and F6 in 3. In 2 strains, the paralell presence of two adhesin types, F4 and F6 was detected. The greatest number of strains 30 (71.42 %) with adhesin F4 belonged to O149 serotype, a considerably smaller number, 4 (26.66%) to serotype O8, 2 strains to serotype O157 and to each serotype O147 and O138 1 strain. The fimbrial adhesin of F5 type was detected in 3 strains which belo

In the present study, isolation of essential oils from leaves of rosemary (Rosmarinus officinalis), sage (Salvia officinalis) and thyme (Thymus vulgaris) by supercritical carbon dioxide as well as hydrodistillation was investigated from the point of yield and antimicrobial activity of obtained extracts against Geobacillus stearotermophillus, Bacillus cereus, Bacillus subtilis var. niger, Enterococcus faecium, Salmonella enteritidis and E.coli strains. Thyme isolates had the highest antibacterial efficiency against tested foodborne bacteria strains (MIC=40-640 μg/cm3) followed by rosemary (MIC=320-1280 μg/cm3) and sage (MIC=160-2560 μg/cm3) isolates. Among tested bacteria, Bacillus species were the most sensitive to presence of all tested supercritical extracts and hydrodistillates. ., U ovoj studiji je ispitana izolacija etarskih ulja iz lista ruzmarina (Rosmarinus officinalis), žalfije (Salvia officinalis) i timijana (Thymus vulgaris) postupkom natkritične ekstrakcije ugljenik(IV)-oksidom sa aspekta prinosa i antimikrobne aktivnosti dobijenih izolata protiv Geobacillus stearotermophillus, Bacillus cereus, Bacillus subtilis var. niger, Enterococcus faecium, Salmonella enteritidis i E.coli sojeva. Izolati timijana su pokazali najjače antibakterijsko dejstvo u odnosu na testirane sojeve (MIC=40-640 μg/cm3). Zatim slede izolati ruzmarina (MIC=320-1280 μg/cm3) i žalfije (MIC=160-2560 μg/cm3). Među testiranim bakterijama, Bacillus sojevi su se pokazali kao najosetljiviji na prisustvo kako natkritičnih ekstrakata, tako i hidrodestilata. .

The multiresistance of bacteria to antibiotics, as well as the lack of new antibiotics on the market encouraged the research of antibacterial activity of non-antibiotic substances including plant extracts. During the previous decades, it has been proven that extracts of certain plants have a strong antibacterial activity, but their clinical use was limited due to the presence of organic solvents. However, plant extracts obtained by the process of supercritical fluid extraction contain no traces of solvents, and the latest researches have established that they do have antibacterial effects on some gram-positive bacteria. This comparative study included extracts of Common Mullein, Angelica and Echinacea obtained by means of supercritical fluid extraction, Soxlet extraction and ultrasound-assisted extraction. The study of their antibacterial activity was performed on some strains of Staphylococcus, Enterobacter cloacae and E. coli isolated from clinical material of human and animal origin. A referential strain of S. aureus ATCC 25923 was included in the research. In the study broth macrodilution method was applied by which the MIC values of extracts were determined. The Angelica extract obtained by ultrasound-assisted extraction had the strongest antibacterial activity, i.e. the lowest MIC value of 40 μg/mL for S. epidermidis strain. The Angelica extract obtained by supercritical fluid extraction also showed substantial antibacterial activity to all Staphylococcus strains included in this study, with the MIC values of 320 to 640 μg/mL. The extracts of Echinacea and Common Mullein obtained by supercritical fluid extraction, as well as of Echinacea extract obtained by Soxlet extraction showed no antibacterial activity since the MIC values of these extracts were 2560 μg/mL or >2560 μg/mL for all bacterial strains icluded in the study., Multirezistencija na antibiotike bakterija kao i nedostatak novih antibiotika na tržištu lekova podstakao je ispitivanja antibakterijskog delovanja supstancija koje nisu antibiotici uključujući i biljne ekstrakte. Tokom prethodnih decenija, dokazano je da ekstrakti pojedinih biljaka imaju izraženo antibakterijsko delovanje ali je zbog prisustva tragova organskih rastvarača, njihova klinička upotreba ogranič ena. Ekstrakti biljaka dobijeni procesom natkritične ekstrakcije, međutim, nemaju tragove rastvarača, a na osnovu dosadašnjih ispitivanja je utvrđeno da deluju na neke gram-pozitivne bakterije. Ovim komparativinim ispitivanjem obuhvaćeni su ekstrakti divizme, anđelike i ehinacee dobijeni procesima natkritične ekstrakcije, Soxlet-ove ekstrakcije i ultrazvučne ekstrakcije. Ispitivanje njihovog antibakterijskog delovanja vršeno je na nekim sojevima stafilokoka, Enterobacter cloacae i E. coli izolovanim iz uzoraka kliničkog materijala poreklom od životinja i ljudi. U ispitivanje je bio uključen i referentni soj S. aureus ATCC 25923. Za ispitivanje antibakterijskog delovanja ekstrakata primenjen je makrodilucioni metod u bujonu pomoću koga su određivane vrednosti MIC ekstrakata. Najjače antibakterijsko delovanje, odnosno najnižu vrednost MIC od 40 μg/mL za soj S. epidermidis imao je ekstrakt anđelike dobijen ultrazvučnom ekstrakcijom. Ekstrakt anđelike dobijen natkritičnom ekstrakcijom je takođe pokazao značajno antibakterijsko delovanje na sve sojeve stafilokoka obuhvaćenih ispitivanjem sa vrednostima MIC od 320 do 640 μg/mL. Ispitivanjem ekstrakata ehinacee i divizme dobijenih procesom nadkritične ekstrakcije kao i ekstrakta ehinacee dobijenog Soxlet-ovom ekstrakcijom, nije utvrđeno antibakterijsko delovanje jer je vrednost MIC ovih ekstrakata iznosila 2560 μg/mL ili >2560 μg/mL za sojeve bakterija koji su obuhvaćeni ispitivanjem.

A prerequisite to the development of an efficient animal health, food safety and traceability management system in the animal food production chain is the implementation of an integrated veterinary informational management system (VIMS) capable for the capture, storage, analysis and retrieval of data and providing the opportunity for the cumulative gathering of the knowledge and capability for its competent interpretation. Such a system will enable collecting appropriate data, including quality management and inspection controls, from all establishments and commodities in the 'from farm to fork' food production chain (farms, holdings, slaughterhouses, laboratories, traders etc.) in a structured, predefined format, and facilitate competent analyses and reporting of such data, as well as the improvement of the existing programs and strategies. The role of information system in animal disease diagnosis, surveillance and notification, control of national and international trade of commodities, food safety management, investigation of diseases, predictive microbiology and quantitative risk assessment is of great importance for the quality of veterinary service. Integral part of the VIMS is animal disease notification system designed according to and in compliance with international requirements, standards and recommendation and able to exchange relevant information with similar information systems. The aim of this contribution is to describe national animal disease notification system which is in place in Serbia as a part of VIMS., Preduslov za uspostavljanje efikasnog sistema menadžmenta zdravstvene zaštite životinja, bezbednosti hrane i sledljivosti u lancu proizvodnje hrane je uvođenje integrisanog veterinarskog informacionog menadžment sistema (VIMS) razvijenog na takav način da omogući prikupljanje, čuvanje, analizu i povlačenje podataka i obezbedi kumulativno skupljanje saznanja i njihovu stručnu interpretaciju. Takav sistem će omogućiti prikupljanje podataka, uključujući one iz upravljanja kvalitetom i u okviru inspekcijskih kontrola, od svih objekata u sistemu proizvodnje hrane 'od farme do viljuške' (farme, gazdinstva, klanice, laboratorije, lica koja se bave prometom i dr.) u strukturirano, unapred definisanom obliku, i obezbediti kompetentnu analizu takvih podataka kao i unapređenje postojećih programa i strategija. Uloga informacionih sistema u dijagnostici, nadzoru i prijavljivanju zaraznih bolesti, kontroli unutrašnjeg i međunarodnog prometa, upravljanju bezbednošću hrane, istraživanju žarišta zaraznih bolesti, prediktivnoj mikrobiologiji i kvantitativnoj analizi rizika je od izuzetnog značaja za kvalitet veterinarske službe. Sastavni deo VIMS-a je sistem za prijavljivanje zaraznih bolesti dizajniran prema i u saglasnosti sa međunarodnim zahtevima, standardima i preporukama i osposobljen da vrši razmenu određenih podatka sa sličnim informacionim sistemima. Cilj ovog rada je da se opiše nacionalni sistem za prijavljivanje zaraznih bolesti koji je u Srbiji u primeni kao deo VIMS-a.

Infections caused by the avian influenza virus have been known for a long time and they are present, to a smaller or greater extent, in both extensive and intensive poultry production in many parts of the world. Epidemiological investigations have established a definite significance of the population of wild birds in maintaining and spreading this infection. Avian influenza is a zoonosis, and the virus has a great potential for causing mortality in humans, in particular its subtypes H5 and H7, which is why it has lately been provoking much attention among scientists and experts, as well as the general public. The objective of the work was to catch a certain number of wild birds in several locations in the Republic of Serbia, to identify them, and to collect samples of their blood serum for the determination of specific antibodies against the avian influenza virus. Birds were caught in ten locations in a manner that was safe for the birds themselves, as well as for the staff that did the catching. The birds were caught in especially produced nets, and in some cases in special traps. The caught wild birds were identified using the methods described in reference literature. All the names of the wild birds were coordinated with the valid Serbian nomenclature of European birds, prepared by prominent ornithologist and bird lover Milan Ružić. Following catching and identification, blood samples were taken from the birds from the wing vein (in bigger birds) or from the leg vein (in smaller birds). In taking blood samples from wild birds, all the principles of asepsis and antisepsis were followed in order to prevent any possibility of infection. After that, the birds were returned to their natural habitat, to the same locations in which they were caught. Serums were isolated from the taken blood samples and they were stored at -20ºC until the final examinations. A total of 46 species of wild birds were identified among a total of 259 birds from which 259 samples of blood ser, Infekcije izazvane virusom avijarne influence, su već odavno poznate i prisutne u manjem ili većem obimu, kako u ekstenzivnoj, tako i u intenzivnoj živinarskoj proizvodnji, u mnogim delovima sveta. Epidemiološkim ispitivanjima je utvrđen nesumnjiv značaj populacije divljih ptica u očuvanju i širenju ove infekcije. Avijarna influenca je zoonoza, a virus ima veliki potencijal da izazove visoku smrtnost kod ljudi, posebno njegovi podtipovi H5 i H7, tako da u novije vreme izaziva veliku pažnju, kako naučne i stručne, tako i najšire javnosti. Cilj ovog rada je bio da se na nekoliko lokacija u Republici Srbiji uhvati određeni broj divljih ptica, izvrši njihova identifikacija i prikupe uzorci krvnog seruma radi otkrivanja specifičnih antitela protiv virusa avijarne influence. Hvatanje ptica vršeno je na deset lokacija na bezbedan način, kako za same ptice, tako i za osoblje koje ih je hvatalo. Hvatanje ptica obavljano je posebnim za te svrhe proizvedenim mrežama, a u nekim slučajevima i posebnim zamkama (klopkama). Za identifikaciju uhvaćenih divljih ptica korišćene su metode koje su opisane u stručnoj literaturi. Svi nazivi divljih ptica usklađeni su sa važećom srpskom nomenklaturom ptica Evrope. Nakon hvatanja i identifikacije, pticama je uzimana krv iz krilne vene (kod većih ptica) ili iz nožne vene (kod malih ptica). Prilikom uzimanja krvi od divljih ptica poštovani su svi principi asepse i antisepse, kako bi se sprečila svaka mogućnost infekcije. Nakon toga, ptice su vraćane u prirodu, na iste lokacije na kojima su i uhvaćene. Od uzetih uzoraka krvi izdvojeni su serumi koji su ostavljani na - 20ºC i čuvani do konačnog ispitivanja. Identifikovano je 46 vrsta divljih ptica sa ukupno 259 jedinki od kojih je izdvojeno 259 uzoraka krvnog seruma. Za otkrivanje prisustva specifičnih antitela protiv virusa avijarne influence u uzorcima krvnog seruma divljih ptica korišćeni su agar gel precipitacija (AGP), test inhibicije hemaglutinacije (IH) za podtipove H5 i H7, cELISA test

Listeria monocytogenes is the causative agent of numerous epidemics and sporadic cases of illness in humans. Food is the principal route of infection. Raw materials of animal and vegetable origin are the potential sources of contamination with this bacterium, particularly the foodstuff undergoing minimal processing procedures. However, in the recent years, emphasis has been increasingly laid on the importance of post-processing contamination occurring through the contact of products with contaminated surfaces in food-processing plants, where this bacterium can persist for months, even for years, as a biofilm. Although it appears to be widely accepted that L. monocytogenes forms biofilms on food-processing surfaces, there has been very little direct microscopical evidence to support this. In this study we investigated the ability of biofilm formation in 14 strains of Listeria monocytogenes of different origin. The biofilms were formed on stainless steel surfaces during 7 days of incubation in a tryptone soy broth supplemented with 0.6% yeast extract (TSB-YE) at a temperature of 25ºC. The developed structures were examined using scanning electron microscopy. Our results revealed significant differences among tested Listeria monocytogenes strains in terms of their ability to form a biofilm in given experimental conditions. Seven of 14 investigated strains of Listeria monocytogenes did not form a biofilm, and only individual bacterial cells were distributed over the stainless steel surface. The strains classified as biofilm producers formed structures of different appearances, from a uniform, confluent monolayer of bacterial cells to individual large, three-dimensional cell aggregates. All strains proved weak producers of extracellular polymeric substance., Listeria monocytogenes je uzročnik brojnih epidemija i sporadičnih slučajeva oboljenja ljudi, a hrana predstavlja glavni put prenošenja infekcije. Sirovine biljnog i animalnog porekla su jedan od mogućih izvora kontaminacije namirnica ovom bakterijom, što se posebno odnosi na prehrambene proizvode koji prolaze minimalni proces obrade. Međutim, poslednjih godina sve više se ističe značaj postprocesne kontaminacije, nastale kao rezultat kontakta prehrambenih proizvoda sa kontaminiranim površinama u pogonima prehrambene industrije, na kojima ova bakterija može u formi biofilma opstati mesecima, pa čak i godinama. Uprkos skoro opšte prihvaćenom stavu da Listeria monocytogenes formira biofilm, malo je mikroskopskih dokaza koji to potvrđuju. U ovom radu ispitana je sposobnost formiranja biofilma kod 14 sojeva Listeria monocytogenes različitog porekla. Biofilmovi su formirani na površini nerđajućeg čelika za sedam dana inkubacije u tripton soja bujonu sa dodatkom 0.6% kvaščevog ekstrakta na temperaturi od 25ºC. Formirane strukture pregledane su skening elektronskom mikroskopijom. Rezultati pregleda potvrđuju da među testiranim sojevima Listeria monocytogenes postoje značajne razlike u sposobnosti da formiraju biofilm pod korišćenim uslovima ispitivanja. Sedam od 14 testiranih sojeva Listeria monocytogenes nije formiralo biofilm, već su se njihove ćelije kao pojedinačne rasporedile po površini nerđajućeg čelika. Sojevi koji su procenjeni kao 'biofilm producers' formirali su strukture različitog izgleda, od uniformnog, konfluentnog monosloja bakterijskih ćelija do pojedinačnih krupnih, trodimenzionalnih ćelijskih agregata. Svi ispitani sojevi su slabo produkovali ekstracelularnu polimeričnu supstanciju.

The resistance of bacteria poses a significant problem everywhere in the world, and consequently in our country as well. The non-critical use of antibacterial medicines in human and veterinary medicine has contributed to the spreading of this resistance. Due to the topical importance of this problem, large numbers of states in the world are financing projects of which the objective is to follow and monitor bacterial resistance. The objective of this investigation was to isolate and identify pathogenic strains of E. coli from piglets with clinically manifest diarrhea and to examine the sensitivity of the isolated strains to a certain number of selected antibiotics. The material for these investigations were parts of intestines (jejunum, ileum) from piglets that died, rectal smears and feces of diseased piglets sampled pig farms in the vicinity of Belgrade. Conventional methods of microbiological diagnostics were used for isolation, and conventional and commercial tests API 20E (bio Merieux, France) were used for identification. Following biochemical identification using hyper immune serums for certain group (O) antigens: (O8, O138, O139, O147, O149, and O157), the serological typization of the strains was carried out. Commercial antiserums: T K88 (F4), K99 (F5), and 987P (F6), Toxigenic E. coli pili antisera, Denka Seiken Co. Ltd. Tokyo, Japan) were used to establish the presence and to identify fimbrial antigens-adhesions through slide agglutination reaction. The sensitivity of the antibiotics to the isolated strains of E. coli was examined using the disc diffusion method according to Kirby Bauer and the microdillution method in bouillon according to CLSI recommendations (2008). Examinations using the microdillution method in bouillon were performed with pure active antibiotic substances: ampicillin, apramycin, gentamicin, kanamycin, tetracycline, ceftriaxone, and ciprofloxacin (Sigma, USA). A total of 400 E. coli strains were isolated, including 48 E. coli strains, Rezistencija bakterija predstavlja značajan problem svuda u svetu, pa i kod nas. Širenju rezistencije doprinela je nekritička upotreba antibakterijskih lekova u humanoj i veterinarskoj medicini. Zbog aktuelnosti problema veliki broj država finansira projekte čiji je cilj praćenje i nadzor nad rezistencijom bakterija. Cilj ovog istraživanja je bio izolacija i identifikacija patogenih sojeva E. coli kod prasadi sa klinički manifestnom dijarejom i ispitivanje osetljivosti izolovanih sojeva na određeni broj odabranih antibiotika. Materijal za ovo ispitivanje predstavljali su delovi creva (jejunum, ileum) uginule prasadi, rektalni brisevi i feces obolele prasadi uzorkovani na farmama svinja iz okoline Beograda. Za izolaciju su korišćene klasične metode mikrobiološke dijagnostike, a za identifikaciju klasični i komercijalni testovi API 20E (bioMerieux, Francuska). Nakon biohemijske identifikacije pomoću hiperimunih seruma za određene grupne (O) antigene (O8, O138, O139, O147, O149 i O157) vršena je serološka tipizacija sojeva. Za utvrđivanje prisustva i identifikaciju fimbrijalnih antigena - adhezina primenom reakcije aglutinacije na pločici su korišćeni komercijalni antiserumi: K88 (F4), K99 (F5) i 987P (F6), Toxigenic E. coli pili antisera, Denka Seiken Co. Ltd. Tokyo, Japan. Osetljivost na antibiotike izolovanih sojeva E. coli ispitivana je primenom disk difuzione metode po Kirby Baueru i mikrodilucione metode u bujonu prema preporukama CLSI (2008). Ispitivanje mikrodilucionom metodom u bujonu je vršeno sa čistim aktivnim supstancama antibiotika: ampicilina, apramicina, gentamicina, kanamicina, tetraciklina, ceftriaksona i ciprofloksacina (Sigma, SAD). Ukupno je izolovano 400 sojeva E. coli, od kojih je 48 svrstano u kategoriju patogenih sojeva. Od navedenog broja patogenih sojeva E. coli kod 32 (66,67%) je ustanovljena multirezistencija na 3 i više od 3 antibiotika, a kod 16 sojeva (33,33%) rezistencija na 2 ili 1 ispitivani antibiotik. Rezistencija na tetraciklin i a

Rezistencija bakterija predstavlja značajan problem svuda u svetu, pa i kod nas. Širenju rezistencije doprinela je nekritička upotreba antibakterijskih lekova u humanoj i veterinarskoj medicini. Zbog aktuelnosti problema veliki broj država finansira projekte čiji je cilj praćenje i nadzor nad rezistencijom bakterija. Cilj ovog istraživanja je bio izolacija i identifikacija patogenih sojeva E. coli kod prasadi sa klinički manifestnom dijarejom i ispitivanje osetljivosti izolovanih sojeva na određeni broj odabranih antibiotika. Materijal za ovo ispitivanje predstavljali su delovi creva (jejunum, ileum) uginule prasadi, rektalni brisevi i feces obolele prasadi uzorkovani na farmama svinja iz okoline Beograda. Za izolaciju su korišćene klasične metode mikrobiološke dijagnostike, a za identifikaciju klasični i komercijalni testovi API 20E (bioMerieux, Francuska). Nakon biohemijske identifikacije pomoću hiperimunih seruma za određene grupne (O) antigene (O8, O138, O139, O147, O149 i O157) vršena je serološka tipizacija sojeva. Za utvrđivanje prisustva i identifikaciju fimbrijalnih antigena - adhezina primenom reakcije aglutinacije na pločici su korišćeni komercijalni antiserumi: K88 (F4), K99 (F5) i 987P (F6), Toxigenic E. coli pili antisera, Denka Seiken Co. Ltd. Tokyo, Japan. Osetljivost na antibiotike izolovanih sojeva E. coli ispitivana je primenom disk difuzione metode po Kirby Baueru i mikrodilucione metode u bujonu prema preporukama CLSI (2008). Ispitivanje mikrodilucionom metodom u bujonu je vršeno sa čistim aktivnim supstancama antibiotika: ampicilina, apramicina, gentamicina, kanamicina, tetraciklina, ceftriaksona i ciprofloksacina (Sigma, SAD). Ukupno je izolovano 400 sojeva E. coli, od kojih je 48 svrstano u kategoriju patogenih sojeva. Od navedenog broja patogenih sojeva E. coli kod 32 (66,67%) je ustanovljena multirezistencija na 3 i više od 3 antibiotika, a kod 16 sojeva (33,33%) rezistencija na 2 ili 1 ispitivani antibiotik. Rezistencija na tetraciklin i a, The resistance of bacteria poses a significant problem everywhere in the world, and consequently in our country as well. The non-critical use of antibacterial medicines in human and veterinary medicine has contributed to the spreading of this resistance. Due to the topical importance of this problem, large numbers of states in the world are financing projects of which the objective is to follow and monitor bacterial resistance. The objective of this investigation was to isolate and identify pathogenic strains of E. coli from piglets with clinically manifest diarrhea and to examine the sensitivity of the isolated strains to a certain number of selected antibiotics. The material for these investigations were parts of intestines (jejunum, ileum) from piglets that died, rectal smears and feces of diseased piglets sampled pig farms in the vicinity of Belgrade. Conventional methods of microbiological diagnostics were used for isolation, and conventional and commercial tests API 20E (bio Merieux, France) were used for identification. Following biochemical identification using hyper immune serums for certain group (O) antigens: (O8, O138, O139, O147, O149, and O157), the serological typization of the strains was carried out. Commercial antiserums: T K88 (F4), K99 (F5), and 987P (F6), Toxigenic E. coli pili antisera, Denka Seiken Co. Ltd. Tokyo, Japan) were used to establish the presence and to identify fimbrial antigens-adhesions through slide agglutination reaction. The sensitivity of the antibiotics to the isolated strains of E. coli was examined using the disc diffusion method according to Kirby Bauer and the microdillution method in bouillon according to CLSI recommendations (2008). Examinations using the microdillution method in bouillon were performed with pure active antibiotic substances: ampicillin, apramycin, gentamicin, kanamycin, tetracycline, ceftriaxone, and ciprofloxacin (Sigma, USA). A total of 400 E. coli strains were isolated, including 48 E. coli strains

The aim of this study was to investigate histomorphometrical characteristics of the thymus, bursa of Fabricius and spleen in the chickens vaccinated with a vaccine against Marek’s disease. For this purpose, we used newly hatched chickens of the light hybrid line, obtained from a local hatchery. The chickens were vaccinated on the 5th day after hatching with a bivalent cell-associated Marek's disease vaccine (PFU-2000 per dose). On day 13 both vaccinated chickens and unvaccinated controls were sacrificed, and thymus, bursa and spleen were removed and processed for light microscopy. The serial tissue sections, hematoxylin-eosine stained, were used for histomorphometric analysis. Vaccination against Marek’s disease decreased the relative mass of the lymphoid organs, and caused significant damage of the thymus and spleen in experimental chicken. In addition, vaccination, similar to Marek's disease virus, induced morphometric changes in the lymphoid organs. Namely, it significantly decreased the diameter and volume of lymphoid follicles, volume of follicular medulla and number of cells in the follicular cortex in the bursa of Fabricius. In the thymus, vaccination reduced the thymus volume and the absolute number of thymocytes. However, vaccination against Marek’s disease caused an increase in the diameter, number and volume of lymphoid follicles in the spleen. The present data suggest that vaccination against Marek’s disease was able to induce the immune response in processed organs, although it reduced the mass and number of lymphocytes in the major lymphoid organs., Cilj ovih ispitivanja su bile histomorfometrijske karakteristike timusa, burze Fabricii i slezine pilića vakcinisanih protiv Marekove bolesti. Za istraživanja su korišćeni jednodnevni pilići lake hibridne linije, gajeni u standardnim uslovima. Pilići su vakcinisani 5. dana od izleganja, bivalentnom vakcinom protiv Marekove bolesti (PFU 2000 po dozi). Trinaestog dana od izleganja kontrolni i vakcinisani pilići su žrtvovani, uzeti su im timus, burza Fabricii i slezina koji su pripremljeni za svetlosno mikroskopsku analizu. Za analizu su korišćeni serijski preseci limfatič nih organa, bojeni metodom hematoksilin-eozin. Dobijeni rezultati su ukazali da vakcinacija protiv Marekove bolesti dovodi do smanjenja relativne mase timusa, burze i slezine, kao i povećanja indeksa oštećenja timusa i slezine. Takođe je ustanovjleno da vakcinacija dovodi do promena morfometrijskih parametara u limfatičnim organima. U burzi Fabricii je uočeno značajno smanjenje dijametra i zapremine limfocitnih folikula, zapremine medule folikula i broja ćelija u korteksu folikula. U timusu vakcinacija dovodi do redukcije zapremine timusa i apsolutnog broja timocita. Međutim, vakcinacija protiv Marekove bolesti u slezini dovodi do povećanja dijametra limfocitnih folikula, kao i do povećanja njihovog broja i zapremine. Rezultati ove studije su ukazali da vakcinacija pilića protiv Marekove bolesti, iako redukuje broj T i B limfocita, indukuje razvoj imunskog odgovora.

Examinations were carried out during the period from January 2006 until March 2007 on a total of 449 samples of the cecum of broilers and the cecum and the colon of pigs. These samples included 251 samples originating from broilers and 198 samples of pig cecums and colons. All the listed samples were obtained by scraping the surface of these parts of the digestive system of broilers and pigs. At the site of sampling, the diluted material was sown on a medium (Karmali agar), in order to get individual colonies. After sowing, the bases were placed in anaerobic jars in which microaerophilic conditions were achieved using Campy Pak, BBL bags. On arrival at the laboratory, the jars containing the sown bases were placed in a thermostat, at a temperature of 42oC for 48 hours for the purpose of incubation. Following incubation, the grown colonies were examined macroscopically, and then microscopic preparations were made from them, which were stained with 2% carbol fuchsin after drying and fixation. Those isolates which were in the form of a comma, the letter S, or gull's wings in the microscopic preparations were considered Campylobacter species (Figures 1 and 2). The isolated thermophilic campylobacteria were identified using conventional and commercial biochemical tests API Campy, manufactured by Bio Mérieux, France. With the application of these microbiological methods, thermophilic Campylobacter species were isolated from 203 (80.88%) of the 251 samples of broiler cecums. Furthermore, thermophilic campylobacteria were isolated from 153 (77.27%) of the 198 samples from the cecum and colon of pigs taken within these investigations. The obtained results indicate that there is a somewhat greater prevalence of these bacteria among the broilers. However, such a high percentage of both broilers and pigs colonized by thermophilic Campylobacter species could pose a serious problem, in particular when it is known that infections of humans caused by the consumption of insufficient, Ispitivanjima obavljenim u periodu od januara 2006. godine do marta 2007. godine ukupno je pregledano 449 uzoraka cekuma brojlera i cekuma i kolona svinja. Od navedenog broja 251 uzorak je poticao od brojlera, a 198 uzoraka iz cekuma i kolona svinja. Svi navedeni uzorci dobijeni su struganjem površine navedenih delova digestivnog sistema brojlera i svinja. Materijal je na mestu uzorkovanja zasejavan sa razređenjem na podlogu (Karmali agar), radi dobijanja pojedinačnih kolonija. Nakon zasejavanja podloge su odlagane u lonce za anaerobe, u kojima su mikroaerofilni uslovi postignuti primenom Campy Pak, BBL kesica. Lonci sa zasejanim podlogama po dopremanju u laboratoriju stavljani su u termostat na temperaturu od 42oC tokom 48 časova radi inkubiranja. Nakon inkubacije izrasle kolonije su pregledane makroskopski, a zatim su od njih pripremani mikroskopski preparati koji su nakon sušenja i fiksiranja bojeni 2% karbol fuksinom. Oni izolati koji su na mikroskopskim preparatima imali oblik zareza, latinskog slova S ili galebovih krila smatrani su Campylobacter vrstama (slika 1 i 2). Identifikacija izolovanih termofilnih kampilobakterija vršena je primenom klasičnih i komercijalnih biohemijskih testova API Campy, proizvođača Bio Mérieux, Francuska. Primenom navedenih mikrobioloških metoda od 251. uzorka iz cekuma brojlera iz 203 (80,88%) izolovane su termofilne Campylobacter vrste. Takođe, od 198 uzoraka iz cekuma i kolona svinja koji su bili obuhvaćeni ispitivanjem termofilne kampilobakterije su izolovane iz 153 (77,27%). Dobijeni rezultati ukazuju na nešto veću prevalenciju ovih bakterija kod brojlera. Međutim, ovako visok procenat kolonizovanih, kako brojlera, tako i svinja termofilnim Campylobacter vrstama može predstavljati ozbiljan problem, posebno ako se zna da infekcije ljudi izazvane konzumiranjem nedovoljno termički obrađenog mesa brojlera i svinja nisu tako retke.

The most reliable diagnosis of an infectuous disease is confirmed by isolation of its pathogen. When it comes to brucellosis, it is important to know that brucella isolation is rarely successful; it is not only very complicated but is as well hazardous for laboratory workers. Due to the above mentioned reasons, it is reasonable to use serological tests for routine diagnosis of this zoonose. This paper deals with examination of bovine sera samples with the aim to detect the titer of specific antibodies against brucellosis. In order to choose and evaluate properly the best test in terms of applicability, speed of performance, and provision of correct results, five serological tests were assayed: rapid serum plate agglutination (Rose Bengal test), Brucella abortus bovis test (RB, BAB test); serum agglutination test (titration) - by Wright, as micro method (mSAT); reaction of complement fixation, and also as micro method (mCF); indirect imunoenzyime test (iELISA) and competitive imunoenzyme test (cELISA). This paper includes 630 samples of bovine blood sera, as well as positive and negative international antibrucella serum as the mandatory control. The presence of specific antibodies against brucella was determined in 125 samples of bovine blood sera. Based on the analysis of the results obtained, evaluation of sensitivity and specificity of these tests was conducted. iELISA and RB test proved to be the most sensitive, while the highest specificity was determined in mCF, and less specific were mSAT and iELISA. RB test had the lowest specificity., Najsigurnija dijagnostika bolesti infektivnog karaktera vrši se izolacijom uzroč nika. Kada je bruceloza u pitanju, mora da se ima u vidu, da je izolacija brucela uspešna u niskom procentu, veoma komplikovana i predstavlja opasnost po laboratorijske radnike. Zbog navedenih razloga, primena seroloških testova u rutinskoj dijagnostici ove zoonoze je opravdana. U ovom radu su vršena ispitivanja uzoraka seruma goveda radi određivanja prisustva titra specifičnih antitela protiv brucela, u svrhu dijagnostikovanja bruceloze. U nameri da se pravilno izbabere i proceni najbolji test u smislu aplikativnosti, brzine izvođenja i dobijanja pouzdanih rezultata, tokom ispitivanja je korišćeno pet seroloških testova: brza serumska aglutinacija na pločici - Rose Bengal test - Brucella abortus bovis test (RB, BAB test); spora aglutinacija (u titraciji) - metoda po Wrightu, kao mikrometoda (mSAT); reakcija vezivanja komplementa, takođe kao mikrometoda (mRVK); indirektni imunoenzimski test (iELISA) i kompetitivni imunoenzimski test (cELISA). Ispitivanjem je bilo obuhvaćeno 630 uzoraka krvnog seruma goveda kao i pozitivni i negativni internacionalni antibrucela serumi kao obavezne kontrole. Prisustvo specifičnih antitela protiv brucela vrsta ustanovljeno je u 125 uzoraka krvnog seruma goveda. Na osnovu analize dobijenih rezultata vršena je procena osetljivosti i specifičnosti navedenih testova. Najosetljivijim su se pokazali iELISA i BAB test, a najveća specifičnost je ustanovljena kod mRVK. Manje specifični bili su mSAT i iELISA, a najmanja specifičnost je ustanovljena kod BAB testa.