Development of PCR-based identification of salmonella enterica serovars

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

Hypodermosis in northern Serbia (Vojvodina)

This paper describes the first documented case of cattle grub (hypodermosis) in Northern Serbia (Vojvodina). Subcutaneous warbles were determined in a six year old Simmental cow, at nine places along the spine. After the extirpation of larvae, based on the morphological characterisation, larvae of the third stage of Hypoderma bovis were diagnosed. The cow was administered therapeutic treatment, which had a favorable outcome, with no signs of recurrence. To the authors best knowledge, the case described in this paper is the first documented case of hypodermosis in cattle in Northern Serbia (Vojvodina). As the climate changed in the past few decades, it is important to pursue detailed investigations of the prevalence of this parasitic myiasis, as there are few such literature data for the Southern region of Serbia. One should also not ignore the fact that species of the genus Hypoderma can cause myiasis in humans as well

Ispitivanje osetljivosti sojeva salmonella vrsta poreklom od živine sa različitih epizootioloških područja na antibiotike

A total of 1666 samples were examined, of which 512 samples of parenchymatous organs of dead or deliberately sacrtificed animals, 60 samples of non-hatched fertilized eggs, 202 samples of feces, 652 samples of cloacal smears, 221 samples of smears from walls of maintenance objects, incubator stations, and transport vehicles, 19 samples of beddings and shavings. The samples originated from poultry farms and which were taken to a laboratory immediately on sampling and sown the same day. A total of 104 strains of Salmonella were isolated: 94 strains from samples of parenchymatous organs of dead chicks, 1 strain from non-hatched eggs, 3 strains from feces samples, 1 strain from samples of cloacal smears, 4 strains from samples of surface smears of maintenance objects and transport vehicles, and 1 strain from samples of beddings and shavings. Serological typization established the presence of the following serovarieties: Salmonella Enteritidis 79 strains, Salmonella Hartford 17 strains, Salmonella Typohimurium 5 strains, Salmonella Mbandaka 2 strains, and Salmonella Glostrup 1 strain. We examined the sensitivity of Salmonella strains to ampicillin, amoxicillin, gentamycin, streptomycin, neomycin, enrofloxacine, norfloxacine, flumequin, erythromycin, lincospectin, colistin, fluorphenicol, and a combination of sulphamethoxasole and trimethoprim. In S. Enteritidis strains, no resistence was established to colistin, fluorphenicol and sulphamethoxasole+trimethoprim, in fact, the sensitivity to these antibiotics and chemotherapeutics was 100%. Prevalence resitence of 0.96%, in only one strain, was established for enrofloxacine. A high prevalence resistence of 33.6% was established for neomycin, while prevalence resistence of 3.86% was established for the related aminoglycozide antibiotic gentamycin. The highest prevalence resistance in S.Hartford strains was established for erythromycin, 15.38%, and streptomycin, 7.6%. Resistence of S. Tyohimurium was established for flumequin and erythromycin in 1.9% strains. No resistance to antibiotics was established in the strains S. Mbandaka and S. Glostrup.Ispitano je ukupno 1666 uzoraka. Od toga 512 uzoraka parenhimatoznih organa uginulih ili prinudno žrtvovanih jedinki, 60 uzoraka oplođenih jaja iz kojih se nisu izlegli pilići (ugušaka), 202 uzorka fecesa, 652 uzorka kloakalnih briseva, 221 uzorak briseva sa zidova objekata za odgoj, inkubatorskih stanica i transportnih vozila i 19 uzoraka prostirke i šuške. Uzorci su poticali sa farmi živine i odmah nakon uzorkovanja dopremani su u laboratoriju i istog dana zasejavani. Ukupno je izolovano 104 soja salmonela: 94 soja iz uzoraka parenhi-matoznih organa uginulih ptica, jedan soj iz uzoraka ugušaka jaja, tri soja iz uzoraka fecesa, jedan soj iz uzoraka kloakalnih briseva, četiri soja iz uzoraka briseva sa površina objekata za odgoj i transportnih vozila i jedan soj iz uzoraka prostirke i šuške. Serološkom tipizacijom ustanovljeno je prisustvo sledećih serovarijeteta: Salmonella Enteritidis 79 sojeva, Salmonella Hartford 17 sojeva, Salmonella Typhimurium pet sojeva, Salmonella Mbandaka dva soja i Salmonella Glostrup jedan soj. Ispitana je osetljivost sojeva Salmonella na ampicilin, amoksicilin, gentamicin, streptomicin, neomicin, enrofloksacin, norfloksacin, flumekvin, eritromicin, linkospektin, kolistin, fluorfenikol i kombinaciju sulfametoksazola i trimetoprima. Kod sojeva S. Enteritidis, nije otkrivena rezistencija na kolistin, fluorfenikol i sulfametoksazol+trimetoprim, odnosno, osetljivost na ove antibakterijske lekove bila je sto posto. Prevalencija rezistencije od 0,96%, odnosno samo kod jednog soja utvrđena je na enrofloksacin. Visoka prevalencija rezistencije od 33,6% ustanovljena je na neomicin, dok je prevalencija rezistencije od 3,86 posto ustanovljena na srodni aminoglikozidni antibiotik gentamicin. Najviša prevalencija rezistencije kod sojeva S. Hartford ustanovljena je na eritromicin od 15,38 posto i streptomicin od 7,6 posto. Rezistencija S. Typhimurium otkrivena je na flumekvin i eritromicin kod 1,9 posto sojeva. Kod sojeva S. Mbandaka i S. Glostrup nije ustanovljena rezistencija na antibiotike

Ispitivanje multirezistencije E.coli i Salmonella koje potiču od domaćih životinja na antibiotike i hemioterapeutike

The presence of multiresistance to the effects of antibiotics and chemotherapeutics and extended spectrum beta-lactamase were examined in 45 strains of E. coli and 35 strains of Salmonella. The strains of E. coli originated from several species of domestic animals: dogs, cats, poultry, and cattle, and 30 strains of Salmonella originated from poultry, 4 strains from cattle, and 1 strain from swine. The presence of the following serovarieties was established using serological examinations: Salmonella Enteritidis 17 strains, Salmonella Gallinarum 1 strain, Salmonella Hartford 5 strains, Salmonella Anatum 1 strain, Salmonella Typhimurium 4 strains, Salmonella Agona 1 strain, Salmonella Infantis 1 strain, Salmonella Thompson var. Berlin 1 strain, Salmonella Tennessee 1 strain, Salmonella Senftenberg 1 strain, Salmonella Glostrup 1 strain, and Salmonella Hadar 1 strain. In the examinations of the listed strains we used antibiogram discs of ampicillin, amoxicillin with clavulanic acid, cephalexin, cephtriaxon, cephotaxim, cephtazidime, aztreonam, gentamycin, chloramphenicol, tetracycline, cyprofloxacine, and a combination of sulphamethoxasole and trimethoprim. The lowest prevalence of multiresistance in E. Coli strains to 3 or more antibiotics was established in dogs 20%, and the highest in 60% strains originating from swine. In 62.88% strains of Salmonella we established sensitivity to all applied antibiotics. Resistance was also established in a small number of the examined strains to ampicillin (11 strains), to tetracycline (5 strains), to amoxicillin with clavulanic acid (5 strains), to sulphamethoxasole with trimethoprim (5 strains), to gentamycin (3 strains), and to cloramphenicol (1 strain). Of all the examined strains of Salmonella, 6 strains originating from poultry exhibited multiresistence. The presence of extended spectrum beta-lactamase effects examined using the ESBL test, was not established in strains of E. coli and Salmonella strains.Ispitivano je prisustvo multirezistencije na antibiotike i hemioterapeutike i beta laktamaze proširenog spektra delovanja kod 45 sojeva E. coli i 35 sojeva Salmonella. Sojevi E. coli su poticali od vise vrsta domaćih životinja: pasa, mačaka, živine i goveda, a 30 sojeva salmonela poticalo je od živine, četiri soja od goveda i jedan soj od svinja. Serološkom determinacijom ustanovljeno je prisustvo navedenih serovarijeteta: Salmonella Enteritidis 17 sojeva, Salmonella Gallinarum 1 soj, Salmonella Hartford 5 sojeva, Salmonella Anatum 1 soj, Salmonella Typhimurium 4 soja, Salmonella Agona 1 soj, Salmonella Infantis 1 soj, Salmonella Thompson var. Berlin 1 soj, Salmonella Tennessee 1 soj, Salmonella Senftenberg 1 soj, Salmonella Glostrup 1 soj i Salmonella Hadar 1 soj. 2a ispitivanje navedenih sojeva su korišćeni antibiogram diskovi ampicilina, amoksicilina sa klavulanskom kiselinom, cefaleksina, ceftriaksona, cefotaksima, ceftazidima, aztreonama, gentamicina, hloramfenikola, tetraciklina, ciprofloksacina i kombinacija sulfametoksazola i trimetoprima. Najniža prevaiencija multirezistencije kod sojeva E. coli na tri ili vise antibiotika ustanovljena je kod pasa 20 posto, a najviša kod 60 posto sojeva koji potiču od svinja. Kod 62,88 posto sojeva salmonela ustanovljena je osetljivost na sve primenjene antibiotike. Rezistencija je utvrđena kod malog broja ispitivanih sojeva i to na ampicilin (11 sojeva), na tetraciklin (5 sojeva), na amoksicilin sa klavulanskom kiselinom (5 sojeva) na sulfametoksazol sa trimetoprimorn (5 sojeva), na gentamicin (3 soja) i na hloramfentkol (1 soj). Od svih ispitanih sojeva salmonela, 6 sojeva koji potiču od živine ispoljilo je multirezistenciju. Prisustvo beta-taktamaza proširenog spektra delovanja ispitanog primenom ESBL testa nije ustanovljeno kod sojeva E. coli i Salmonella

Krpeljska fauna autohtone pramenke u Istočnoj Srbiji

Sheep production is an example of a sustainable production fully integrated within the local rural development. One of the main threats on the outdoor breeding of sheep is parasitism. Ticks are nuisances and vectors of several diseases agents. The distribution of ticks appears to be changing, with spread to previously unaffected areas. Tick and tick-borne disease control is one of the major components of animal health program protecting livestock in the developing countries, which reflects impact on the livelihood of resource-poor farming communities. Taking into consideration the negative impact on the health status of the livestock, also the direct and indirect economic losses, it is necessary to examine the tolerance and resistance of certain species against diseases. It is one of the most important elements of the strategy of selection and screening for resistant animals. The aim of this study was to determine the tick species persisting in 45 tested autochthonous Zackel sheep flocks, and examine their seasonal occurrence from March 2010 to January 2011, in the region of South Serbia. The result showed that Ixodes, Dermacentor, Ripicephalus and Haemaphysalis were the most abundant ticks found, affecting 50.40% tested sheep. The result of this study is a survey of tick species from autochthonous Zackel sheep in Serbia and implication of possible preventions measures for diseases caused and transmitted by ticks.Ovčarstvo predstavlja primer održive proizvodnje koja čini sastavni deo ruralnog razvoja. Jedan od glavnih zdravstvenih problema kod ekstenzivnog načina uzgoja ovaca predstavljaju parazitske infekcije. Krpelji su vektori za uzročnike mnogobrojnih oboljenja. Rasprostranjenost krpelja se menja i u novije vreme ih nalazimo i na novim arealima. U zemljama u razvoju, borba protiv krpelja i oboljenja prenosica krpeljima predstavljaju jedan od glavnih strateških tačaka zdravstvenog nadzora nad životinjama i ljudima. Uzimajući u obzir značaj direktnih i indirektnih ekonomskih gubitaka izazvanih krpeljima i oboljenjima čiji su oni uzročnici, posebna pažnja treba da se posveti ispitivanju tolerancije i otpornosti ka parazitskim bolestima pojedinih vrsta i rasa životinja. Potraga za otpornim jedinkama i njihova selekcija treba da bude deo strategije stočarstva. Cilj ovog rada je bio da prikaže rezultate ispitivanja o prisustvu krpeljske faune u 45 zapata ovaca autohtone pramenke. Ispitana je sezonalna distribucija pojave pojedinih krpelja u periodu između marta 2010. i januara 2011. godine, u regionu Istočne Srbije. Kod 50, 40% ispitanih ovaca ustanovljeno je prisustvo krpelja. Rezultati pokazuju da su krpelji iz rodova Ixodes, Dermacentor, Ripicephalus i Haemaphysalis najučestaliji u zapatima ovaca autohtone pramenke

Case Report of dirofilariasis in grey wolf in Serbia

This paper describes a case of dirofilariasis in a two-year old, female grey wolf (Canis lupus lupus). The autopsy revealed the presence of 42 adult forms of Dirofilaria immitis in the pulmonary artery, right ventricle and right atrium, varying in length from 9.5 to 30 cm. This is, to the best of our knowledge, the second report of D. immitis in grey wolves in Serbia. Our finding confirms that the wolf, as a subspecies distinct from the dog, should also be considered as a very suitable definitive host for dirofilariasis

Cytological and bacteriological testing of infertile mares in Vojvodina district

SUMMARY Background: While inflammation of the endometrium (endomertitis) is among the leading reproductive disorders in mares, it is often neglected during the therapy of their infertility. Ultrasonography, as well as microbiological and cytological investigations are the most frequently applied techniques in the diagnosis of endometritis. However, in most cases a single method is not efficient and the proper diagnosis requires the simultaneous evaluation of the results of more of them.objectives: The authors’ aim was to assess the reproductive disorders of mares in the Vojvodina region, paying special attention to the endometritis incidence rate, possible causes, as well as to the applicability of the diagnostic methods available in field conditions.Materials and methods: 21 infertile mares were included in the investigation. Their uterine swabs were taken with a double protected cytobrush catheter. The obtained samples were undergone to bacteriological and cytological testing. Results and discussion: Cytological findings revealed severe inflammation in 23.8 % (n = 5) of the cases (more than 5 PMN/HPF ) and Str. equi spp. zooepidemicus was isolated from all of these samples. Moderate inflammation (2–5 PMN/HPF) was cytologically detected in 28.5% (n = 6) of the mares. In this group 9.5% (n = 2) of them were bacteriologically negative, Corynebacterium strains were isolated at the same ratio, while E. coli and Pseudomonas sp. were both found in 4.7% (n = 1). According to our investigation, simultaneous cytological and bacteriological testing of uterine samples gives more accurate information on the cause of endometritis than both methods separately. The fact that 80% of the samples were bacteriologically positive proves that inflammation in the uterus is among the main causes of infertility in mares

Uticaj zamrzavanja ribe na odabrane parametre kvaliteta dimljenog proizvoda

In the circumstances of small processing facilities for smoked fish production, and a particularly good fishing season, there occurs the necessity for freezing the catch in order to process it and preserve it in this way, at times when the season is not so good. To this end, the goal of our research was to examine the effect of freezing fish on the selected parameters of cold smoked, vacuum-packed trout fillets. To conduct the experiment, two groups of trout were formed: the control one, out of fresh fish, and the experimental one, out of previously frozen fish. In the course of a three week period of smoked fish storage at the temperature of 4ºC the variation of the total number of bacteria, and the total number of lactobacilli were monitored. Also, at the end of processing, the content value of water and salt were determined, as well as the content value of salt in the water phase, and aw value (water activity). By testing the bacteriological status of two groups of products during the storage period, it was determined that the total number of both bacteria and lactobacilli was smaller in the samples produced from frozen fish. The average content value of water and NaCl, as well as the salt content value in the water phase, in the vacuum-packed, smoked fillets produced from frozen fish was statistically, considerably larger than the content value of water, salt, and salt content value in the water phase in the samples manufactured from fresh fish. Our research findings show that using the raw material that was previously frozen for producing smoked fish fillets is not only appropriate, but also recommended.U uslovima malih preradnih kapaciteta za proizvodnju dimljene ribe nameće se potreba, u sezonama većeg izlova, za zamrzavanjem ribe i njena dalja prerada u vremenu kada je taj izlov mali, ili ga nema. Zbog toga je cilj našeg istraživanja bio da se ispita uticaj zamrzavanja ribe na odabrane parametre kvaliteta hladno dimljenih vakuumiranih fileta pastrmke. Za eksperiment su formirane dve grupe pastrmki: kontrolna od sveže ribe i ogledna od prethodno zamrznute ribe. U toku tri nedelje skladištenja proizvoda dimljene ribe pri temperaturi od 4ºC praćena je promena ukupnog broja bakterija i ukupan broj laktobacila, a na kraju proizvodnje utvrđen je sadržaj vode i soli, izračunat je sadržaj soli u vodenoj fazi i aw vrednost (aktivnost vode). Ispitivanjem bakteriološkog statusa obe grupe proizvoda, utvrđeno je da je i ukupan broj bakterija i ukupan broj laktobacila u uzorcima, tokom skladištenja, bio manji kod uzoraka proizvedenih od zamrzavane ribe. Prosečan sadržaj vode, NaCl, kao i sadržaj soli u vodenoj fazi u vakuumiranim dimljenim filetima proizvedenim od zamrznute ribe bio je statistički značajno veći od sadržaja vode, soli i sadržaja soli u vodenoj fazi u poređenju sa uzorcima proizvedenim od sveže ribe. Rezultati naših istraživanja ukazuju da je opravdano, čak i poželjno, u proizvodnji dimljenih fileta ribe koristiti sirovinu koja je prethodno bila zamrznuta

Bakterijska mikroflora pluća imunokompromitovanih prasadi sa pneumonijom

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