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In September 2018, a four-month-old dog with fever and petechial bleeding came to the internal clinic at the Faculty of Veterinary Medicine University of Belgrade. On hematology analysis, thrombocytopenia and mild anemia were observed. Examination of the blood smear revealed platelet inclusions. The commercial serology test was positive for Anaplasma spp. The dog was treated with doxycycline for 14 days, and after 48 hours from the beginning of the treatment, the symptoms subsided. PCR analysis and sequencing confirmed infection with A. platys., U septembru 2018. godine, štene staro četiri meseca, sa znacima groznice i petehijalnim krvarenjem, je došlo u ambulantu Klinike za male životinje Fakulteta veterinarske medicine Univerziteta u Beogradu. Hematološka analiza je pokazala trombocitopeniju i blagu anemiju. Pregledom krvnog razmaza ustanovljene su inkluzije u trombocitima. Komercijalni serološki test je pokazao prisustvo antitela protiv Anaplasma spp. Sprovedena je terapija doksiciklinom u trajanju od 14 dana, a posle 48 sati od početka terapije znaci bolesti su se povukli. PCR analizom i senkvencioniranjem je potvrđeno prisustvo Anaplasma platys.

In September 2018, a four-month-old dog with fever and petechial bleeding came to the internal clinic at the Faculty of Veterinary Medicine University of Belgrade. On hematology analysis, thrombocytopenia and mild anemia were observed. Examination of the blood smear revealed platelet inclusions. The commercial serology test was positive for Anaplasma spp. The dog was treated with doxycycline for 14 days, and after 48 hours from the beginning of the treatment, the symptoms subsided. PCR analysis and sequencing confirmed infection with A. platys., U septembru 2018. godine, štene staro četiri meseca, sa znacima groznice i petehijalnim krvarenjem, je došlo u ambulantu Klinike za male životinje Fakulteta veterinarske medicine Univerziteta u Beogradu. Hematološka analiza je pokazala trombocitopeniju i blagu anemiju. Pregledom krvnog razmaza ustanovljene su inkluzije u trombocitima. Komercijalni serološki test je pokazao prisustvo antitela protiv Anaplasma spp. Sprovedena je terapija doksiciklinom u trajanju od 14 dana, a posle 48 sati od početka terapije znaci bolesti su se povukli. PCR analizom i senkvencioniranjem je potvrđeno prisustvo Anaplasma platys.

In September 2018, a four-month-old dog with fever and petechial bleeding came to the internal clinic at the Faculty of Veterinary Medicine University of Belgrade. On hematology analysis, thrombocytopenia and mild anemia were observed. Examination of the blood smear revealed platelet inclusions. The commercial serology test was positive for Anaplasma spp. The dog was treated with doxycycline for 14 days, and after 48 hours from the beginning of the treatment, the symptoms subsided. PCR analysis and sequencing confirmed infection with A. platys., U septembru 2018. godine, štene staro četiri meseca, sa znacima groznice i petehijalnim krvarenjem, je došlo u ambulantu Klinike za male životinje Fakulteta veterinarske medicine Univerziteta u Beogradu. Hematološka analiza je pokazala trombocitopeniju i blagu anemiju. Pregledom krvnog razmaza ustanovljene su inkluzije u trombocitima. Komercijalni serološki test je pokazao prisustvo antitela protiv Anaplasma spp. Sprovedena je terapija doksiciklinom u trajanju od 14 dana, a posle 48 sati od početka terapije znaci bolesti su se povukli. PCR analizom i senkvencioniranjem je potvrđeno prisustvo Anaplasma platys.

In September 2018, a four-month-old dog with fever and petechial bleeding came to the internal clinic at the Faculty of Veterinary Medicine University of Belgrade. On hematology analysis, thrombocytopenia and mild anemia were observed. Examination of the blood smear revealed platelet inclusions. The commercial serology test was positive for Anaplasma spp. The dog was treated with doxycycline for 14 days, and after 48 hours from the beginning of the treatment, the symptoms subsided. PCR analysis and sequencing confirmed infection with A. platys., U septembru 2018. godine, štene staro četiri meseca, sa znacima groznice i petehijalnim krvarenjem, je došlo u ambulantu Klinike za male životinje Fakulteta veterinarske medicine Univerziteta u Beogradu. Hematološka analiza je pokazala trombocitopeniju i blagu anemiju. Pregledom krvnog razmaza ustanovljene su inkluzije u trombocitima. Komercijalni serološki test je pokazao prisustvo antitela protiv Anaplasma spp. Sprovedena je terapija doksiciklinom u trajanju od 14 dana, a posle 48 sati od početka terapije znaci bolesti su se povukli. PCR analizom i senkvencioniranjem je potvrđeno prisustvo Anaplasma platys.

In September 2018, a four-month-old dog with fever and petechial bleeding came to the internal clinic at the Faculty of Veterinary Medicine University of Belgrade. On hematology analysis, thrombocytopenia and mild anemia were observed. Examination of the blood smear revealed platelet inclusions. The commercial serology test was positive for Anaplasma spp. The dog was treated with doxycycline for 14 days, and after 48 hours from the beginning of the treatment, the symptoms subsided. PCR analysis and sequencing confirmed infection with A. platys., U septembru 2018. godine, štene staro četiri meseca, sa znacima groznice i petehijalnim krvarenjem, je došlo u ambulantu Klinike za male životinje Fakulteta veterinarske medicine Univerziteta u Beogradu. Hematološka analiza je pokazala trombocitopeniju i blagu anemiju. Pregledom krvnog razmaza ustanovljene su inkluzije u trombocitima. Komercijalni serološki test je pokazao prisustvo antitela protiv Anaplasma spp. Sprovedena je terapija doksiciklinom u trajanju od 14 dana, a posle 48 sati od početka terapije znaci bolesti su se povukli. PCR analizom i senkvencioniranjem je potvrđeno prisustvo Anaplasma platys.

Q fever is a zoonotic disease caused by Coxiella burnetii, a gram-negative coccobacillus, which has been detected in a wide range of animal species, mostly domestic ruminants, but also in wild mammals, pets, birds, reptiles, arthropods (especially ticks), as well as in humans. Although the exposure to domestic animals in rural areas is regarded as the most common cause of the disease in humans, recent studies have shown that the role of pets in the epidemiology of Q fever has been increasingly growing. Although the primary route of infection is inhalation, it is presumed that among animals the infection circulates through ticks and that they are responsible for heterospecific transmission, as well as spatial dispersion among vertebrates. The aim of this study was to determine the presence and prevalence of C. burnetii in ticks removed from stray dogs, as well as to examine the distribution of tick species parasitizing dogs on the territory of Belgrade city. A PCR protocol targeting IS1111 repetitive transposon-like region of C. burnetii was used for the detection of C. burnetii DNA in ticks and the results were confirmed by sequence analysis. In total, 316 ticks were collected from 51 stray dogs - 40 females (78.43%) and 11 males (21.57%). Three species of ticks were identified: Rhipicephalus sanguineus (72.15%), Ixodes ricinus (27.53%) and Dermacentor reticulatus (0.32%). Out of 316 examined ticks, C. burnetii DNA was detected only in the brown dog tick R. sanguineus, with a total prevalence of 10.53% (24/228). The high prevalence of C. burnetii in R. sanguineus, which is primarily a dog tick, indicates the importance of dogs in the epidemiology of Q fever in the territory of Belgrade.

Q fever is a zoonotic disease caused by Coxiella burnetii, a gram-negative coccobacillus, which has been detected in a wide range of animal species, mostly domestic ruminants, but also in wild mammals, pets, birds, reptiles, arthropods (especially ticks), as well as in humans. Although the exposure to domestic animals in rural areas is regarded as the most common cause of the disease in humans, recent studies have shown that the role of pets in the epidemiology of Q fever has been increasingly growing. Although the primary route of infection is inhalation, it is presumed that among animals the infection circulates through ticks and that they are responsible for heterospecific transmission, as well as spatial dispersion among vertebrates. The aim of this study was to determine the presence and prevalence of C. burnetii in ticks removed from stray dogs, as well as to examine the distribution of tick species parasitizing dogs on the territory of Belgrade city. A PCR protocol targeting IS1111 repetitive transposon-like region of C. burnetii was used for the detection of C. burnetii DNA in ticks and the results were confirmed by sequence analysis. In total, 316 ticks were collected from 51 stray dogs - 40 females (78.43%) and 11 males (21.57%). Three species of ticks were identified: Rhipicephalus sanguineus (72.15%), Ixodes ricinus (27.53%) and Dermacentor reticulatus (0.32%). Out of 316 examined ticks, C. burnetii DNA was detected only in the brown dog tick R. sanguineus, with a total prevalence of 10.53% (24/228). The high prevalence of C. burnetii in R. sanguineus, which is primarily a dog tick, indicates the importance of dogs in the epidemiology of Q fever in the territory of Belgrade.

The goal of this study was to compare the results of serological and conventional bacteriological methods with the results obtained using multiplex PCR Bruce-ladder assay. Based on the obtained results, the usability of the assay was assessed in regard to rapid diagnosis of canine brucellosis directly from the samples of reproductive organs of infected dogs. Out of 225 blood samples, 33 (14.67%) had a positive agglutination reaction. In this study, out of the 225 assayed reproductive organs of dogs, B. canis was isolated from 3 samples (1.33%), while the PCR Bruce-ladder assay detected two positive samples (0.88%). Two dogs from which B. canis was isolated, an antibody titer of 1/200 was established in blood serums, and third dog from which B. canis was isolated was negative using the tube agglutination test. From a total of 225 assayed organ samples, a positive PCR reaction was obtained from two samples. The obtained results show that the tube agglutination method remains the first choice for the detection of dogs infected with B. canis. In addition, whenever possible, it is necessary to try isolation. It is desirable to attempt the detection of B. canis in tissues using PCR, but the results may not be treated as definitive and reliable.

The aim of the study was to evaluate and adapt the PCR-based protocol that utilizes the developed serotype-specific primers to identify Salmonella enterica species and its serotypes that are most frequently isolated from poultry samples in Vojvodina. Using the slide agglutination test, 64 and 33 out of 107 Salmonella isolates were identified as S. Infantis and S. Enteritidis, respectively, while ten isolates were identified as eight different Salmonella serovars. Using the same isolates, presence of 993-bp (bcfC gene), 636-bp (steB gene) and 293-bp (sdf locus) amplicons in multiplex PCR unambiguously identified 31 isolates as S. Enteritidis. Two isolates identified as Enteritidis in slide agglutination test were not identified as such in PCR-based approach since they both were missing 293bp long PCR product. Thirty-nine isolates produced a 727-bp amplicon in the specific simplex PCR, and thus were identified as S. Infantis. The greatest discrepancy in comparison to the results of conventional serotyping has been observed in the case of S. Infantis, since 25 more isolates were noted as S. Infantis by conventional serotyping. Seven isolates, with unexpected PCR profiles stayed unidentified by molecular typing, although they were serotyped as S. Typhimurium (1) and S. Infantis (6). S. Gallinarum serovar has to be additionally confirmed, since it shares the same PCR profile with S. Livingstone. Clearly, PCR-based identification has to be thoroughly checked, verified and adapted if it is to be applied as the routine identification protocol.

The aim of the study was to evaluate and adapt the PCR-based protocol that utilizes the developed serotype-specific primers to identify Salmonella enterica species and its serotypes that are most frequently isolated from poultry samples in Vojvodina. Using the slide agglutination test, 64 and 33 out of 107 Salmonella isolates were identified as S. Infantis and S. Enteritidis, respectively, while ten isolates were identified as eight different Salmonella serovars. Using the same isolates, presence of 993-bp (bcfC gene), 636-bp (steB gene) and 293-bp (sdf locus) amplicons in multiplex PCR unambiguously identified 31 isolates as S. Enteritidis. Two isolates identified as Enteritidis in slide agglutination test were not identified as such in PCR-based approach since they both were missing 293bp long PCR product. Thirty-nine isolates produced a 727-bp amplicon in the specific simplex PCR, and thus were identified as S. Infantis. The greatest discrepancy in comparison to the results of conventional serotyping has been observed in the case of S. Infantis, since 25 more isolates were noted as S. Infantis by conventional serotyping. Seven isolates, with unexpected PCR profiles stayed unidentified by molecular typing, although they were serotyped as S. Typhimurium (1) and S. Infantis (6). S. Gallinarum serovar has to be additionally confirmed, since it shares the same PCR profile with S. Livingstone. Clearly, PCR-based identification has to be thoroughly checked, verified and adapted if it is to be applied as the routine identification protocol.

The goal of this study was to compare the results of serological and conventional bacteriological methods with the results obtained using multiplex PCR Bruce-ladder assay. Based on the obtained results, the usability of the assay was assessed in regard to rapid diagnosis of canine brucellosis directly from the samples of reproductive organs of infected dogs. Out of 225 blood samples, 33 (14.67%) had a positive agglutination reaction. In this study, out of the 225 assayed reproductive organs of dogs, B. canis was isolated from 3 samples (1.33%), while the PCR Bruce-ladder assay detected two positive samples (0.88%). Two dogs from which B. canis was isolated, an antibody titer of 1/200 was established in blood serums, and third dog from which B. canis was isolated was negative using the tube agglutination test. From a total of 225 assayed organ samples, a positive PCR reaction was obtained from two samples. The obtained results show that the tube agglutination method remains the first choice for the detection of dogs infected with B. canis. In addition, whenever possible, it is necessary to try isolation. It is desirable to attempt the detection of B. canis in tissues using PCR, but the results may not be treated as definitive and reliable.

For the purpose of precise antibiotic susceptibility testing it is necessary to clearly distinguish Pseudomonas and Stenotrophomonas genera, considering acquired resistance of Pseudomonas species, as well as the intrinsic resistance of Stenotrophomonas species. This is why in the identification of the 51 isolates originated from fish, the following methods were used: standard PCR, 16S rRNA gene sequencing, and MALDI-TOF. The results of the standard PCR test, 16S rRNA gene sequencing and MALDI-TOF analysis confirmed 35 strains to belong to the Pseudomonas genus. Standard PCR test and VITEK MS device confirmed that 10 strains belong to Stenotrophomonas maltophilia species. Three strains were positive in both standard PCR tests for Pseudomonas and Stenotrpohomonas. 16S rRNA gene sequencing identified these 3 strains to be 99% Pseudomonas sp. and 99% Stenotrophomonas sp. VITEK MS first identified these three strains as 99% Stenotrophomonas, and in the repeated identification it identified them as 99% Pseudomonas. MALDI TOF/TOF 4800 Plus device identified these strains as Stenotrophomonas. Three strains were negative in both standard PCR tests for Pseudomonas and Stenotrpohomonas. 16S rRNA gene sequencing identified these 3 strains to be 99% Pseudomonas sp. and 99% Stenotrophomonas sp. VITEK MS first identified these three strains as 99% Stenotrophomonas, and in the repeated identification it identified them as 99% Pseudomonas. MALDI TOF/TOF 4800 Plus device identified these strains as Stenotrophomonas. Although modern test methods that have very high specificity (PCR, 16S rRNA gene Pseudomonas and Stenotrophomonas species for 6 isolates could not be reached using the above mentioned methods., S obzirom na značaj stečene rezistencije Pseudomonas vrsta, kao i na intrinzičnu rezistenciju Stenotrophomonas vrsta, a u cilju preciznog ispitivanja osetljivosti na antibiotike, neophodna je jasna diferencijacija pripadnika ovih rodova bakterija. U tom cilju su u identifikaciji 51 izolata poreklom od riba korišćene metode: standardni PCR, 16S rRNA sekvenciranje gena, MALDI-TOF. Rezultati standardnog PCR testa, 16S rRNA sekvenciranja gena i MALDI-TOF analize su za 35 sojeva potvrdili pripadnost rodu Pseudomonas. Standardnim PCR testom i primenom aparata VITEK MS utvrđeno je da 10 sojeva pripada vrsti Stenotrophomonas maltophilia. U 16S rRNA sekvenciranju gena 3 soja koja su bila pozitivna u oba standardna PCR testa identifikovana su kao 99% Pseudomonas sp. i 99% Stenotrophomonas sp. VITEK MS je ova tri soja u prvoj identifikaciji identifikovao kao 99% Stenotrophomonas, a u ponovljenoj identifikaciji kao 99% Pseudomonas. Ti sojevi su na aparatu MALDI TOF/TOF 4800 Plus bili identifikovani kao Stenotrophomonas. U 16S rRNA sekvenciranju gena tri soja koja su bila negativna u oba standardna PCR testa su identifikovana kao 99% Pseudomonas sp. i 99% Stenotrophomonas sp. Aparat VITEK MS je ova tri soja identifikovao u jednoj identifikaciji kao 99% Stenotrophomonas, a u drugoj identifikaciji 99% kao Pseudomonas. Ti sojevi su na aparatu MALDI TOF/ TOF 4800 Plus bili identifikovani kao Stenotrophomonas. Iako su u ovom istraživanju korišćene savremene metode ispitivanja koje imaju vrlo visoku specifičnost (PCR, 16s rRNK sequencing, MALDI TOF) precizna diferencijacija Pseudomonas i Stenotrophomonas vrsta nije mogla biti postignuta.

For the purpose of precise antibiotic susceptibility testing it is necessary to clearly distinguish Pseudomonas and Stenotrophomonas genera, considering acquired resistance of Pseudomonas species, as well as the intrinsic resistance of Stenotrophomonas species. This is why in the identification of the 51 isolates originated from fish, the following methods were used: standard PCR, 16S rRNA gene sequencing, and MALDI-TOF. The results of the standard PCR test, 16S rRNA gene sequencing and MALDI-TOF analysis confirmed 35 strains to belong to the Pseudomonas genus. Standard PCR test and VITEK MS device confirmed that 10 strains belong to Stenotrophomonas maltophilia species. Three strains were positive in both standard PCR tests for Pseudomonas and Stenotrpohomonas. 16S rRNA gene sequencing identified these 3 strains to be 99% Pseudomonas sp. and 99% Stenotrophomonas sp. VITEK MS first identified these three strains as 99% Stenotrophomonas, and in the repeated identification it identified them as 99% Pseudomonas. MALDI TOF/TOF 4800 Plus device identified these strains as Stenotrophomonas. Three strains were negative in both standard PCR tests for Pseudomonas and Stenotrpohomonas. 16S rRNA gene sequencing identified these 3 strains to be 99% Pseudomonas sp. and 99% Stenotrophomonas sp. VITEK MS first identified these three strains as 99% Stenotrophomonas, and in the repeated identification it identified them as 99% Pseudomonas. MALDI TOF/TOF 4800 Plus device identified these strains as Stenotrophomonas. Although modern test methods that have very high specificity (PCR, 16S rRNA gene Pseudomonas and Stenotrophomonas species for 6 isolates could not be reached using the above mentioned methods., S obzirom na značaj stečene rezistencije Pseudomonas vrsta, kao i na intrinzičnu rezistenciju Stenotrophomonas vrsta, a u cilju preciznog ispitivanja osetljivosti na antibiotike, neophodna je jasna diferencijacija pripadnika ovih rodova bakterija. U tom cilju su u identifikaciji 51 izolata poreklom od riba korišćene metode: standardni PCR, 16S rRNA sekvenciranje gena, MALDI-TOF. Rezultati standardnog PCR testa, 16S rRNA sekvenciranja gena i MALDI-TOF analize su za 35 sojeva potvrdili pripadnost rodu Pseudomonas. Standardnim PCR testom i primenom aparata VITEK MS utvrđeno je da 10 sojeva pripada vrsti Stenotrophomonas maltophilia. U 16S rRNA sekvenciranju gena 3 soja koja su bila pozitivna u oba standardna PCR testa identifikovana su kao 99% Pseudomonas sp. i 99% Stenotrophomonas sp. VITEK MS je ova tri soja u prvoj identifikaciji identifikovao kao 99% Stenotrophomonas, a u ponovljenoj identifikaciji kao 99% Pseudomonas. Ti sojevi su na aparatu MALDI TOF/TOF 4800 Plus bili identifikovani kao Stenotrophomonas. U 16S rRNA sekvenciranju gena tri soja koja su bila negativna u oba standardna PCR testa su identifikovana kao 99% Pseudomonas sp. i 99% Stenotrophomonas sp. Aparat VITEK MS je ova tri soja identifikovao u jednoj identifikaciji kao 99% Stenotrophomonas, a u drugoj identifikaciji 99% kao Pseudomonas. Ti sojevi su na aparatu MALDI TOF/ TOF 4800 Plus bili identifikovani kao Stenotrophomonas. Iako su u ovom istraživanju korišćene savremene metode ispitivanja koje imaju vrlo visoku specifičnost (PCR, 16s rRNK sequencing, MALDI TOF) precizna diferencijacija Pseudomonas i Stenotrophomonas vrsta nije mogla biti postignuta.

For the purpose of precise antibiotic susceptibility testing it is necessary to clearly distinguish Pseudomonas and Stenotrophomonas genera, considering acquired resistance of Pseudomonas species, as well as the intrinsic resistance of Stenotrophomonas species. This is why in the identification of the 51 isolates originated from fish, the following methods were used: standard PCR, 16S rRNA gene sequencing, and MALDI-TOF. The results of the standard PCR test, 16S rRNA gene sequencing and MALDI-TOF analysis confirmed 35 strains to belong to the Pseudomonas genus. Standard PCR test and VITEK MS device confirmed that 10 strains belong to Stenotrophomonas maltophilia species. Three strains were positive in both standard PCR tests for Pseudomonas and Stenotrpohomonas. 16S rRNA gene sequencing identified these 3 strains to be 99% Pseudomonas sp. and 99% Stenotrophomonas sp. VITEK MS first identified these three strains as 99% Stenotrophomonas, and in the repeated identification it identified them as 99% Pseudomonas. MALDI TOF/TOF 4800 Plus device identified these strains as Stenotrophomonas. Three strains were negative in both standard PCR tests for Pseudomonas and Stenotrpohomonas. 16S rRNA gene sequencing identified these 3 strains to be 99% Pseudomonas sp. and 99% Stenotrophomonas sp. VITEK MS first identified these three strains as 99% Stenotrophomonas, and in the repeated identification it identified them as 99% Pseudomonas. MALDI TOF/TOF 4800 Plus device identified these strains as Stenotrophomonas. Although modern test methods that have very high specificity (PCR, 16S rRNA gene Pseudomonas and Stenotrophomonas species for 6 isolates could not be reached using the above mentioned methods., S obzirom na značaj stečene rezistencije Pseudomonas vrsta, kao i na intrinzičnu rezistenciju Stenotrophomonas vrsta, a u cilju preciznog ispitivanja osetljivosti na antibiotike, neophodna je jasna diferencijacija pripadnika ovih rodova bakterija. U tom cilju su u identifikaciji 51 izolata poreklom od riba korišćene metode: standardni PCR, 16S rRNA sekvenciranje gena, MALDI-TOF. Rezultati standardnog PCR testa, 16S rRNA sekvenciranja gena i MALDI-TOF analize su za 35 sojeva potvrdili pripadnost rodu Pseudomonas. Standardnim PCR testom i primenom aparata VITEK MS utvrđeno je da 10 sojeva pripada vrsti Stenotrophomonas maltophilia. U 16S rRNA sekvenciranju gena 3 soja koja su bila pozitivna u oba standardna PCR testa identifikovana su kao 99% Pseudomonas sp. i 99% Stenotrophomonas sp. VITEK MS je ova tri soja u prvoj identifikaciji identifikovao kao 99% Stenotrophomonas, a u ponovljenoj identifikaciji kao 99% Pseudomonas. Ti sojevi su na aparatu MALDI TOF/TOF 4800 Plus bili identifikovani kao Stenotrophomonas. U 16S rRNA sekvenciranju gena tri soja koja su bila negativna u oba standardna PCR testa su identifikovana kao 99% Pseudomonas sp. i 99% Stenotrophomonas sp. Aparat VITEK MS je ova tri soja identifikovao u jednoj identifikaciji kao 99% Stenotrophomonas, a u drugoj identifikaciji 99% kao Pseudomonas. Ti sojevi su na aparatu MALDI TOF/ TOF 4800 Plus bili identifikovani kao Stenotrophomonas. Iako su u ovom istraživanju korišćene savremene metode ispitivanja koje imaju vrlo visoku specifičnost (PCR, 16s rRNK sequencing, MALDI TOF) precizna diferencijacija Pseudomonas i Stenotrophomonas vrsta nije mogla biti postignuta.

The aim of this study was to investigate the influence of marination, on the growth of Salmonella spp. in contaminated broiler breast fillets during storage. In the conducted study, broiler breast fillets were inoculated with a cocktail of different Salmonella strains and afterwards marinated in different solutions of table salt, sodium tripolyphosphate and/or sodium citrate. The total count of Salmonella spp. was examined on the 0, 3rd, 6th and 9th day of storage. Broiler breast fillets salted in 6% solution of table salt were used as the control. Proximate composition and salt content, pH value and a(w) value, were determined as the meat quality parameters and parameters which can affect environmental conditions for bacterial growth, as well. Compared to initial contamination, Salmonella spp. count in marinated and salted fillets did not change significantly (p < 0.001) until the 3rd and 6th day of storage, respectively, but it increased significantly on the 9th day of storage. Marination of broiler breast fillets in different solutions of table salt, sodium tripolyphosphate and/or sodium citrate had a significant influence(p < 0.05; p < 0.01) on pH and a(w) value, moisture, ash and salt content in marinated broiler meat, but there was no significant influence (p > 0.05) on protein and fat content in broiler meat. According to the results obtained it can be concluded that marination of broiler breast fillets in solutions containing table salt, sodium trpolyphosphate and/or citrate, in some way, can prolongate the lag phase of Salmonella spp. growth, where sodium citrate is more effective than sodium tripolyphosphate.

The aim of this study was to investigate the influence of marination, on the growth of Salmonella spp. in contaminated broiler breast fillets during storage. In the conducted study, broiler breast fillets were inoculated with a cocktail of different Salmonella strains and afterwards marinated in different solutions of table salt, sodium tripolyphosphate and/or sodium citrate. The total count of Salmonella spp. was examined on the 0, 3rd, 6th and 9th day of storage. Broiler breast fillets salted in 6% solution of table salt were used as the control. Proximate composition and salt content, pH value and a(w) value, were determined as the meat quality parameters and parameters which can affect environmental conditions for bacterial growth, as well. Compared to initial contamination, Salmonella spp. count in marinated and salted fillets did not change significantly (p < 0.001) until the 3rd and 6th day of storage, respectively, but it increased significantly on the 9th day of storage. Marination of broiler breast fillets in different solutions of table salt, sodium tripolyphosphate and/or sodium citrate had a significant influence(p < 0.05; p < 0.01) on pH and a(w) value, moisture, ash and salt content in marinated broiler meat, but there was no significant influence (p > 0.05) on protein and fat content in broiler meat. According to the results obtained it can be concluded that marination of broiler breast fillets in solutions containing table salt, sodium trpolyphosphate and/or citrate, in some way, can prolongate the lag phase of Salmonella spp. growth, where sodium citrate is more effective than sodium tripolyphosphate.

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.

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 mu g/mL to >64 mu g/mL and cefoxitin MIC values ranged from 1 pg/mL to >256 mu g/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.

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

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

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

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

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

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

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.

Bacterial strains which possess genes to produce ESBL most often are multiresistant and also carry genes responsible for the resistance to most other antibiotics, including aminoglycosides, sulfamethoxazole+trimethoprim and fluoroquinolones. Therefore, practically the biggest contemporary clinical problem are infections of humans and animals caused by ESBL-producing strains of E. coli, Kleibsiella, Enterobacter, Proteus, Serratia, Citrobacter, Salmonella and Shigella species. The investigation of the ESBL presence was completed on multiresistant E. coli and Salmonella strains originating from dogs, cats, cattle, sheep, goats, pigs and poultry. The investigated strains were isolated from ear, skin, vaginal, faecal, urine, egg and eggshell swabs, from healthy and diseased individual animals of various ages and breed categories. The sum of 112 E. coli and 45 Salmonella strains was investigated. All strains resistant to 3 or more antibiotics were categorized as multiresistant, which led to a conclusion that 35 E. coli and 6 Salmonella strains out of all investigated were multiresistant to antibiotics. The largest number of multiresistant E. coli strains was discovered in cattle - 12 in total, and the minimal number in goats and sheep, with two strains each. All multiresistant Salmonella strains belonged to the Salmonella Enteritidis species (S. enterica subsp. enterica serovar Enteritidis). The sum of multiresistant Salmonella strains compared to all investigated strains was relatively low (13.3%), but the resistance prevalence for some antibiotics in these strains was extremely high, for ampicillin and amoxicillin with clavulanic acid as high as 100%, and for tetracycline 83.3%. For the control in this investigation were used ESBL positive E. coli strains originated in human urine specimens. No presence of positive ESBL strains was established. However, when the screening investigation was performed, almost all the strains were suspect, thus a confirmatory test had to, Sojevi bakterija koji poseduju gene za proizvodnju beta-laktamaza proširenog spektra delovanja (ESBL) najčešće ispoljavaju multirezistenciju i istovremeno su nosioci gena odgovornih za rezistenciju na većinu drugih antibiotika, uključujući i aminoglikozide, sulfametoksazol+trimetoprim i fluorohinolone. Zbog navedenih razloga, najveći problem danas u kliničkoj praksi predstavljaju infekcije ljudi i životinja izazvane ESBL produkujućim sojevima E.coli, Klebsiella, Enterobacter, Proteus, Serratia, Citrobacter, Salmonella i Shigella vrsta. U ovim ispitivanjima materijali za izolovanje sojeva bakterija predstavljali su brisevi ušiju, kože, vaginalni brisevi, feces i urin, jaja i ljuske od jaja. Primenom klasičnih i komercijalnih testova mikrobiološke dijagnostike izolovani su i identifikovani sojevi E.coli i Salmonella vrsta. Navedenim sojevima je ispitivana i osetljivost na određeni broj antibiotika i hemioterapeutika primenom metoda propisanih od strane CLSI, USA iz 2006. Samo oni sojevi E.coli i Salmonella poreklom od pasa, mačaka, goveda, ovaca, koza, svinja i živine kod kojih je ustanovljena rezistencija na 3 i više antibiotika kategorizovani su kao multirezistentni i dalje su ispitivani na prisustvo betalaktamaza proširenog spektra delovanja. Ovim ispitivanjem obuhvaćeno je ukupno 112 sojeva E.coli i 45 sojeva Salmonella. Takođe su ispitivanjem bila obuhvać ena i dva ESBL pozitivna soja E.coli koji su izolovani u našoj laboratoriji iz urina ljudi, a služili su nam kao kontrola. Od 112 ispitivanih sojeva E.coli, kod 35 je utvrđena multirezistencija na određeni broj antibiotika, dok je od 45 sojeva Salmonella koji su ispitivani samo kod 6 sojeva ustanovljena multirezistencija. Najveći broj multirezistetnih sojeva E.coli (MR E.coli) otkriven je kod goveda, ukupno 12, a najmanje kod koza i ovaca, po dva soja. Svi multirezistentni sojevi Salmonella pripadali su vrsti Salmonella enteritidis (S.enterica subsp. enterica serovar. enteritidis). Ukupan broj multirezistentnih

Listeria monocytogenes is the causative agent of listeriosis in humans and animals and an important food-born pathogen. Control of its presence in food processing plants, particularly on sites where food contamination is expected, is of paramount importance with respect to food safety and protection of human health. Numerous studies demonstrated that this organism can be isolated during several months, even several years, from diverse sites in food processing plants, which is due to its ability to adsorb onto inert surfaces and form a biofilm, alone or in coexistence with other bacterial species. In this study we investigated the ability of 16 animal isolates of L. monocytogenes to form a biofilm on polystyrene microtiter plates. The investigation was performed at three different temperatures 4oC, 25 oC and 37oC that are commonly suitable for growth of Listeria monocytogenes. The research was carried out using three different nutritive media: trypthon-soy broth with yeast extract (TSB-YE), brainheart infusion (BHI) and 1/20 diluted trypthon-soy broth with yeast extract (1/20 TSB-YE). In order to investigate the biofilm formation in vitro an inoculum was prepared from 24-hours-old cultures of isolated strains of L.monocytogenes. The density of the inoculum was 2-10x107 cfu/mL (OD600=0.093 ± 0.009 in TSB-YE). The microtiter plates were incubated at cited temperatures during 48h. Colonization rate of L.monocytogenes strains on polystyrene surface and biofilm formation were monitored using crystal violet stain added to the microtiter plates, as well as using light microscopy. The tested strains demonstrated a diverse ability of biofilm formation, depending on the incubation temperature and nutritive medium. In this paper we presented the results obtained for four strains of Listeria monocytogenes isolated from brain samples of sheep, designated as 785/05, 593/05, 748/05, 1915/04 and one strain isolated from aborted calve's fetus, designated as 021/04. The referent strai, Listeria monocytogenes je uzročnik listerioze ljudi i životinja i veoma važan patogen koji pored ostalih načina prenošenja može da se prenese i hranom. Kon- trola njenog prisustva u pogonima prehrambene industrije, naročito na mestima moguće kontaminacije namirnica, od izuzetnog je značaja u proizvodnji zdravstveno bezbedne hrane i zaštiti zdravlja ljudi. Istraživanja su pokazala da se ova bakterija više meseci ili godina uzastopno može izolovati sa različitih mesta sa kojima u kontakt dolaze namirnice, upravo zahvaljujući sposobnosti da se vezuje za inertne površine i formira biofilm, sama ili u zajednici sa drugim vrstama bakterija. U ovom radu je ispitivana sposobnost formiranja biofilma u mikrotitracionim pločama od polistirena kod 16 izolovanih sojeva L. monocytogenes, poreklom od životinja. Ispitivanja su vršena na temperaturama od 4oC, 25oC i 37oC, na kojima Listeria monocytogenes može uobičajeno da raste. Za navedena ispitivanja korišč ene su tri hranljive podloge različitog sastava, kao što su tripton soja bujon sa kvaščevim ekstraktom (TSB-YE), moždano srčana infuzija (BHI) i tripton soja bujon sa kvaščevim ekstraktom razređen u odnosu 1:20 fiziološkim rastvorom (1/20 TSB-YE). Za ispitivanja formiranja biofilma in vitro pripreman je inokulum od kultura izolovanih sojeva L. monocytogenes starih 24 časa. Gustina pripremljenog inokuluma iznosila je 2-10x107 CFU/ml (OD600=0,093 ± 0,009 u TSB-YE). Mikrotitracione ploče nakon inokulisanja inkubisane su tokom 48 časova na prethodno navedenim temperaturama. Kolonizacija sojeva L. monocytogenes na polistiren i formiranje biofilma, praćeno je pomoću kristal violet boje koja je dodavana u mikrotitracione ploče, kao i pomoću svetlosnog mikroskopa. Sojevi Listeria monocytogenes koji su ispitivani pokazali su različitu sposobnost formiranja biofilma u zavisnosti od temperature inkubacije i podloge koja je korišćena za rast. Zbog navedenih činjenica u radu su prikazani rezultati dobijeni ispitivanjem četiri soja Listeria

Listeria monocytogenes is the causative agent of listeriosis in humans and animals and an important food-born pathogen. Control of its presence in food processing plants, particularly on sites where food contamination is expected, is of paramount importance with respect to food safety and protection of human health. Numerous studies demonstrated that this organism can be isolated during several months, even several years, from diverse sites in food processing plants, which is due to its ability to adsorb onto inert surfaces and form a biofilm, alone or in coexistence with other bacterial species. In this study we investigated the ability of 16 animal isolates of L. monocytogenes to form a biofilm on polystyrene microtiter plates. The investigation was performed at three different temperatures 4oC, 25 oC and 37oC that are commonly suitable for growth of Listeria monocytogenes. The research was carried out using three different nutritive media: trypthon-soy broth with yeast extract (TSB-YE), brainheart infusion (BHI) and 1/20 diluted trypthon-soy broth with yeast extract (1/20 TSB-YE). In order to investigate the biofilm formation in vitro an inoculum was prepared from 24-hours-old cultures of isolated strains of L.monocytogenes. The density of the inoculum was 2-10x107 cfu/mL (OD600=0.093 ± 0.009 in TSB-YE). The microtiter plates were incubated at cited temperatures during 48h. Colonization rate of L.monocytogenes strains on polystyrene surface and biofilm formation were monitored using crystal violet stain added to the microtiter plates, as well as using light microscopy. The tested strains demonstrated a diverse ability of biofilm formation, depending on the incubation temperature and nutritive medium. In this paper we presented the results obtained for four strains of Listeria monocytogenes isolated from brain samples of sheep, designated as 785/05, 593/05, 748/05, 1915/04 and one strain isolated from aborted calve's fetus, designated as 021/04. The referent strai, Listeria monocytogenes je uzročnik listerioze ljudi i životinja i veoma važan patogen koji pored ostalih načina prenošenja može da se prenese i hranom. Kon- trola njenog prisustva u pogonima prehrambene industrije, naročito na mestima moguće kontaminacije namirnica, od izuzetnog je značaja u proizvodnji zdravstveno bezbedne hrane i zaštiti zdravlja ljudi. Istraživanja su pokazala da se ova bakterija više meseci ili godina uzastopno može izolovati sa različitih mesta sa kojima u kontakt dolaze namirnice, upravo zahvaljujući sposobnosti da se vezuje za inertne površine i formira biofilm, sama ili u zajednici sa drugim vrstama bakterija. U ovom radu je ispitivana sposobnost formiranja biofilma u mikrotitracionim pločama od polistirena kod 16 izolovanih sojeva L. monocytogenes, poreklom od životinja. Ispitivanja su vršena na temperaturama od 4oC, 25oC i 37oC, na kojima Listeria monocytogenes može uobičajeno da raste. Za navedena ispitivanja korišč ene su tri hranljive podloge različitog sastava, kao što su tripton soja bujon sa kvaščevim ekstraktom (TSB-YE), moždano srčana infuzija (BHI) i tripton soja bujon sa kvaščevim ekstraktom razređen u odnosu 1:20 fiziološkim rastvorom (1/20 TSB-YE). Za ispitivanja formiranja biofilma in vitro pripreman je inokulum od kultura izolovanih sojeva L. monocytogenes starih 24 časa. Gustina pripremljenog inokuluma iznosila je 2-10x107 CFU/ml (OD600=0,093 ± 0,009 u TSB-YE). Mikrotitracione ploče nakon inokulisanja inkubisane su tokom 48 časova na prethodno navedenim temperaturama. Kolonizacija sojeva L. monocytogenes na polistiren i formiranje biofilma, praćeno je pomoću kristal violet boje koja je dodavana u mikrotitracione ploče, kao i pomoću svetlosnog mikroskopa. Sojevi Listeria monocytogenes koji su ispitivani pokazali su različitu sposobnost formiranja biofilma u zavisnosti od temperature inkubacije i podloge koja je korišćena za rast. Zbog navedenih činjenica u radu su prikazani rezultati dobijeni ispitivanjem četiri soja Listeria

Bacterial strains which possess genes to produce ESBL most often are multiresistant and also carry genes responsible for the resistance to most other antibiotics, including aminoglycosides, sulfamethoxazole+trimethoprim and fluoroquinolones. Therefore, practically the biggest contemporary clinical problem are infections of humans and animals caused by ESBL-producing strains of E. coli, Kleibsiella, Enterobacter, Proteus, Serratia, Citrobacter, Salmonella and Shigella species. The investigation of the ESBL presence was completed on multiresistant E. coli and Salmonella strains originating from dogs, cats, cattle, sheep, goats, pigs and poultry. The investigated strains were isolated from ear, skin, vaginal, faecal, urine, egg and eggshell swabs, from healthy and diseased individual animals of various ages and breed categories. The sum of 112 E. coli and 45 Salmonella strains was investigated. All strains resistant to 3 or more antibiotics were categorized as multiresistant, which led to a conclusion that 35 E. coli and 6 Salmonella strains out of all investigated were multiresistant to antibiotics. The largest number of multiresistant E. coli strains was discovered in cattle - 12 in total, and the minimal number in goats and sheep, with two strains each. All multiresistant Salmonella strains belonged to the Salmonella Enteritidis species (S. enterica subsp. enterica serovar Enteritidis). The sum of multiresistant Salmonella strains compared to all investigated strains was relatively low (13.3%), but the resistance prevalence for some antibiotics in these strains was extremely high, for ampicillin and amoxicillin with clavulanic acid as high as 100%, and for tetracycline 83.3%. For the control in this investigation were used ESBL positive E. coli strains originated in human urine specimens. No presence of positive ESBL strains was established. However, when the screening investigation was performed, almost all the strains were suspect, thus a confirmatory test had to, Sojevi bakterija koji poseduju gene za proizvodnju beta-laktamaza proširenog spektra delovanja (ESBL) najčešće ispoljavaju multirezistenciju i istovremeno su nosioci gena odgovornih za rezistenciju na većinu drugih antibiotika, uključujući i aminoglikozide, sulfametoksazol+trimetoprim i fluorohinolone. Zbog navedenih razloga, najveći problem danas u kliničkoj praksi predstavljaju infekcije ljudi i životinja izazvane ESBL produkujućim sojevima E.coli, Klebsiella, Enterobacter, Proteus, Serratia, Citrobacter, Salmonella i Shigella vrsta. U ovim ispitivanjima materijali za izolovanje sojeva bakterija predstavljali su brisevi ušiju, kože, vaginalni brisevi, feces i urin, jaja i ljuske od jaja. Primenom klasičnih i komercijalnih testova mikrobiološke dijagnostike izolovani su i identifikovani sojevi E.coli i Salmonella vrsta. Navedenim sojevima je ispitivana i osetljivost na određeni broj antibiotika i hemioterapeutika primenom metoda propisanih od strane CLSI, USA iz 2006. Samo oni sojevi E.coli i Salmonella poreklom od pasa, mačaka, goveda, ovaca, koza, svinja i živine kod kojih je ustanovljena rezistencija na 3 i više antibiotika kategorizovani su kao multirezistentni i dalje su ispitivani na prisustvo betalaktamaza proširenog spektra delovanja. Ovim ispitivanjem obuhvaćeno je ukupno 112 sojeva E.coli i 45 sojeva Salmonella. Takođe su ispitivanjem bila obuhvać ena i dva ESBL pozitivna soja E.coli koji su izolovani u našoj laboratoriji iz urina ljudi, a služili su nam kao kontrola. Od 112 ispitivanih sojeva E.coli, kod 35 je utvrđena multirezistencija na određeni broj antibiotika, dok je od 45 sojeva Salmonella koji su ispitivani samo kod 6 sojeva ustanovljena multirezistencija. Najveći broj multirezistetnih sojeva E.coli (MR E.coli) otkriven je kod goveda, ukupno 12, a najmanje kod koza i ovaca, po dva soja. Svi multirezistentni sojevi Salmonella pripadali su vrsti Salmonella enteritidis (S.enterica subsp. enterica serovar. enteritidis). Ukupan broj multirezistentnih

The investigation of the presence of plasmids was performed on E. coli and Salmonella strains, originating from dogs, cats, cattle, pigs and poultry. The investigated bacterial strains were isolated from ear and skin swabs vaginal swabs, faeces and urine, obtained from healthy and diseased animals of various age and breed categories. Up to now 45 E. coli and 35 Salmonella strains have been isolated, but the presence of plasmids was investigated in 42 strains of bacteria resistant to at least 3 or more antibiotics (23 E. coli and 4 Salmonella strains). The plasmids were isolated from 2 E. coli strains originating from dogs, 4 E. coli strains originating from cattle, 2 E. coli strains originating from pigs, 1 E. coli strain originating from a cat and 2 Salmonella strains originating from poultry. The obtained approximate size values of isolated plasmids from investigated strains expressed in base pairs were from 2,4 kb to 17,5 kb. After the transformation process was completed and the E. coli DH 5α transformants streaking was done on LB agar with added antibiotics, the visible growth of transformed bacteria meant that the competent cells have received DNA (plasmid) with the resistance gene for the antibiotic. Lack of growth in transformants meant that the resistance gene was to be found on the chromosome., Ispitivanje prisustva plazmida vršeno je na sojevima E. coli i Salmonella vrsta poreklom od pasa, mačaka, goveda, svinja i živine. Ispitivani sojevi bakterija izolovani su iz briseva ušiju, kože, vaginalnih briseva, fecesa i urina i to kako od zdravih tako i od bolesnih jedinki različitih starosnih kategorija i rasa. Ukupno je izolovano 45 sojeva E. coli i 35 sojeva Salmonella vrsta a prisustvo plazmida ispitano je kod 42 soja bakterija rezistentnih na najmanje 3 i više antibiotika. Plazmidi su izolovani iz 11 ispitanih sojeva i to iz 2 soja E. coli poreklom od pasa, 4 soja E. coli poreklom od goveda, 2 soja E. coli poreklom od svinja, 1 soja E. coli poreklom od mačke i kod 2 soja Salmonella vrsta poreklom od živine. Nakon izvršene transformacije i zasejavanja transformanata E. coli DH 5α na LB agar sa dodatkom antibiotika, vidljivi porast transformisanih bakterija značio je da su kompetentne ćelije primile DNK (plazmid) sa genom rezistencije na taj antibiotik. Izostanak porasta transformanata značio je da se gen rezistencije nalazio na hromozomu.

Being an important pathogenic and indicator microorganism, E. coli has been included in all the resistance follow up programs worldwide. We carried out sampling on 42 cattle, pig and poultry farms. After sampling the material was delivered to the laboratories, where it was exposed to standard treatment and streaking by standard methods of microbiological diagnostics with the aim to isolate and identify E. coli strains. On the selected isolated E. coli strains an investigation of its sensitivity to a certain number of antimicrobial agents (ampicillin, tetracycline, chloramphenicol streptomycin, neomycin and sulphametoxasol) was carried out. After obtaining the results from each farm for the three different animals categories (young healthy, young unhealthy, adult), 60 E. coli strains for eachcategori were isolated to be examined for sensitivity to 17 antibiotics and chemotherapeutics. The investigation included a total of 180 E. coli strains originating from cattle, pigs and poultry. For the investigation of isolated E. coli strains Mueller Hinton broth and agar, antibiogram tablets (Torlak Belgrade) as well as discs by Becton Dickinson, BBL (USA) were used In isolated E.coli strains resistance to investigated antimicrobial agents was significantly different depending on the species age and health status of the animals. The majority of the isolated E. coli strains examined for 17 antibiotics and chemotherapeutics showed a resistance to 15, and sensitivity only to cephalosporines of the III generation and colistin., U svetu je E.coli kao važan patogeni i indikatorski mikroorganizam uključena u sve programe praćenja rezistencije i upravo iz tih razloga je i u našim uslovima sprovedeno uzorkovanje materijala na 42 farme goveda, svinja i živine. Materijal je po uzorkovanju dopreman do laboratorije nakon čega je vršena obrada i zasejavanje primenom uobičajenih metoda mikrobiološke dijagnostike u cilju izolacije i identifikacije sojeva E.coli. Na odabranim izolovanim sojevima E. coli izvršeno je ispitivanje osetljivosti rezistencije na sledeća antimikrobna sredstva: ampicilin, tetraciklin hloramfenikol, streptomicin, neomicin i sulfametoksazol. Nakon dobijanja rezultata sa svake farme iz tri različite kategorije životinja (mlade zdrave, mlade bolesne, odrasle) izdvojeno je po 60 sojeva E.coli za ispitivanje osetljivosti - rezistencije na 17 antibiotika i hemioterapeutika. Ispitivanjem je obuhvaćeno ukupno 180 sojeva E. coli poreklom od goveda, svinja i živine. U ispitivanju su korišćeni Mueller Hinton agar, antibiogram tablete i diskovi proizvođača Torlak i Becton Dickinson, BBL (USA). Kod izolovanih sojeva E.coli prevalencija rezistencije na ispitivana antimikrobna sredstva značajno se razlikovala u zavisnosti od vrste, starosti i zdravstvenog stanja životinja. Kod većeg broja izolovanih sojeva E.coli ispitanih na 17 antibiotika i hemio­terapeutika ustanovljena je rezistencija na 15 a osetljivost samo na cefalosporine III generacije i kolistin.

Being an important pathogenic and indicator microorganism, E. coli has been included in all the resistance follow up programs worldwide. We carried out sampling on 42 cattle, pig and poultry farms. After sampling the material was delivered to the laboratories, where it was exposed to standard treatment and streaking by standard methods of microbiological diagnostics with the aim to isolate and identify E. coli strains. On the selected isolated E. coli strains an investigation of its sensitivity to a certain number of antimicrobial agents (ampicillin, tetracycline, chloramphenicol streptomycin, neomycin and sulphametoxasol) was carried out. After obtaining the results from each farm for the three different animals categories (young healthy, young unhealthy, adult), 60 E. coli strains for eachcategori were isolated to be examined for sensitivity to 17 antibiotics and chemotherapeutics. The investigation included a total of 180 E. coli strains originating from cattle, pigs and poultry. For the investigation of isolated E. coli strains Mueller Hinton broth and agar, antibiogram tablets (Torlak Belgrade) as well as discs by Becton Dickinson, BBL (USA) were used In isolated E.coli strains resistance to investigated antimicrobial agents was significantly different depending on the species age and health status of the animals. The majority of the isolated E. coli strains examined for 17 antibiotics and chemotherapeutics showed a resistance to 15, and sensitivity only to cephalosporines of the III generation and colistin., U svetu je E.coli kao važan patogeni i indikatorski mikroorganizam uključena u sve programe praćenja rezistencije i upravo iz tih razloga je i u našim uslovima sprovedeno uzorkovanje materijala na 42 farme goveda, svinja i živine. Materijal je po uzorkovanju dopreman do laboratorije nakon čega je vršena obrada i zasejavanje primenom uobičajenih metoda mikrobiološke dijagnostike u cilju izolacije i identifikacije sojeva E.coli. Na odabranim izolovanim sojevima E. coli izvršeno je ispitivanje osetljivosti rezistencije na sledeća antimikrobna sredstva: ampicilin, tetraciklin hloramfenikol, streptomicin, neomicin i sulfametoksazol. Nakon dobijanja rezultata sa svake farme iz tri različite kategorije životinja (mlade zdrave, mlade bolesne, odrasle) izdvojeno je po 60 sojeva E.coli za ispitivanje osetljivosti - rezistencije na 17 antibiotika i hemioterapeutika. Ispitivanjem je obuhvaćeno ukupno 180 sojeva E. coli poreklom od goveda, svinja i živine. U ispitivanju su korišćeni Mueller Hinton agar, antibiogram tablete i diskovi proizvođača Torlak i Becton Dickinson, BBL (USA). Kod izolovanih sojeva E.coli prevalencija rezistencije na ispitivana antimikrobna sredstva značajno se razlikovala u zavisnosti od vrste, starosti i zdravstvenog stanja životinja. Kod većeg broja izolovanih sojeva E.coli ispitanih na 17 antibiotika i hemio­terapeutika ustanovljena je rezistencija na 15 a osetljivost samo na cefalosporine III generacije i kolistin.

Being an important pathogenic and indicator microorganism, E. coli has been included in all the resistance follow up programs worldwide. We carried out sampling on 42 cattle, pig and poultry farms. After sampling the material was delivered to the laboratories, where it was exposed to standard treatment and streaking by standard methods of microbiological diagnostics with the aim to isolate and identify E. coli strains. On the selected isolated E. coli strains an investigation of its sensitivity to a certain number of antimicrobial agents (ampicillin, tetracycline, chloramphenicol streptomycin, neomycin and sulphametoxasol) was carried out. After obtaining the results from each farm for the three different animals categories (young healthy, young unhealthy, adult), 60 E. coli strains for eachcategori were isolated to be examined for sensitivity to 17 antibiotics and chemotherapeutics. The investigation included a total of 180 E. coli strains originating from cattle, pigs and poultry. For the investigation of isolated E. coli strains Mueller Hinton broth and agar, antibiogram tablets (Torlak Belgrade) as well as discs by Becton Dickinson, BBL (USA) were used In isolated E.coli strains resistance to investigated antimicrobial agents was significantly different depending on the species age and health status of the animals. The majority of the isolated E. coli strains examined for 17 antibiotics and chemotherapeutics showed a resistance to 15, and sensitivity only to cephalosporines of the III generation and colistin., U svetu je E.coli kao važan patogeni i indikatorski mikroorganizam uključena u sve programe praćenja rezistencije i upravo iz tih razloga je i u našim uslovima sprovedeno uzorkovanje materijala na 42 farme goveda, svinja i živine. Materijal je po uzorkovanju dopreman do laboratorije nakon čega je vršena obrada i zasejavanje primenom uobičajenih metoda mikrobiološke dijagnostike u cilju izolacije i identifikacije sojeva E.coli. Na odabranim izolovanim sojevima E. coli izvršeno je ispitivanje osetljivosti rezistencije na sledeća antimikrobna sredstva: ampicilin, tetraciklin hloramfenikol, streptomicin, neomicin i sulfametoksazol. Nakon dobijanja rezultata sa svake farme iz tri različite kategorije životinja (mlade zdrave, mlade bolesne, odrasle) izdvojeno je po 60 sojeva E.coli za ispitivanje osetljivosti - rezistencije na 17 antibiotika i hemioterapeutika. Ispitivanjem je obuhvaćeno ukupno 180 sojeva E. coli poreklom od goveda, svinja i živine. U ispitivanju su korišćeni Mueller Hinton agar, antibiogram tablete i diskovi proizvođača Torlak i Becton Dickinson, BBL (USA). Kod izolovanih sojeva E.coli prevalencija rezistencije na ispitivana antimikrobna sredstva značajno se razlikovala u zavisnosti od vrste, starosti i zdravstvenog stanja životinja. Kod većeg broja izolovanih sojeva E.coli ispitanih na 17 antibiotika i hemio­terapeutika ustanovljena je rezistencija na 15 a osetljivost samo na cefalosporine III generacije i kolistin.