Tick-Borne Encephalitis in Sheep, Romania.

Little is known about the occurrence of tick-borne encephalitis in Romania. Sheep are an infection source for humans and are useful sentinels for risk analysis. We demonstrate high antibody prevalence (15.02%) among sheep used as sentinels for this disease in 80% of the tested localities in 5 counties of northwestern Romania.

Intentional fatal metallic phosphide poisoning in a dog--a case report.

BACKGROUND: Metallic phosphides are extremely toxic pesticides that are regulated in their usage. Information concerning the impact of metallic phosphides on human health is abundant. Data regarding the clinical pathology of phosphide poisoning in humans or domestic and wild animals is largely incomplete with only a few cases of metallic phosphide poisoning being reported every year, especially in humans. For the majority of cases reported in dogs the data are vague or incomplete. Here we report a complete and detailed description of pathological changes in a case of intentional metallic phosphide poisoning in a dog including an exhaustive examination of the brain. CASE PRESENTATION: A 1 year old, male, Belgian Shepherd crossbreed dog with a clean medical history and no observed clinical signs prior to death, was submitted for post mortem examination. The dog was found dead by the owner. Near the body a suspect mix of bread, fat and a blackish powder was found. The owner announced the authorities and submitted the animal and the possible bait for forensic examination. At necropsy, multisystemic necrotic and degenerative lesions were observed. Histological exam confirmed the presence of necrotic and degenerative lesions of variable severity in all of the examined organs. The toxicological forensic examination revealed the presence of the phosphine gas in the gastric content and the bait. CONCLUSION: Metallic phosphide poisoning is a rarely reported entity, since the diagnosis of intentional poisoning with these compounds is a great challenge for forensic pathologists and toxicologists. To our knowledge, this is the first study describing the lesions completely in veterinary forensic toxicology. We assume that the toxic shows systemic endotheliotropism and damage of the endothelial cells responsible for the hemorrhagic lesions and for the secondary ischemic necrosis in various organs. This report will contribute to a better understanding of the pathogenesis in cases of acute metallic phosphide exposure in animals.

Congenital diaphragmatic hernia with concurrent aplasia of the pericardium in a foal.

BACKGROUND: In veterinary medicine congenital abnormalities of the diaphragm and pericardium are rare, idiopathic malformations, being reported mainly in dogs. This report documents an unusual case of developmental defects in a foal consisting of diaphragmatic hernia concurrent with pericardial aplasia. CASE PRESENTATION: Following a normal delivery, a full term, female Friesian stillborn foal with the placenta was presented for necropsy. External morphological examination indicated a normally developed foal. At necropsy, a large oval defect (approximately 20 × 15 cm in size) was observed in the left-dorsal side of the diaphragm (left lumbocostal triangle). This defect allowed the intestinal loops, spleen and partially the liver to translocate into the thorax. The loops of the left ascending colon, including the pelvic flexure and partially the small intestine covered the cranial and dorsal posterior parts of the heart due to the complete absence of the left pericardium. The remaining pericardium presented as a white, semi-transparent strip, partially covering the right side of the heart. The left lung and the main bronchus were severely hypoplastic to approximately one-fifth the size of their right homologue. The intermediate part of the liver, containing mainly the enlarged quadrate lobe was translocated in the thorax, severely enlarged and showed marked fibrosis. Histologically in the herniated lobes we diagnosed hepatic chronic passive congestion, telangiectasia and medial hypertrophy of blood vessels. CONCLUSION: Concomitant malformation involving diaphragmatic hernia and pericardial aplasia in horses have not been previously reported. Moreover, this is the first case describing pericardial aplasia in horse.

Prevalence and Antimicrobial Resistance of Escherichia coli Isolates from Chicken Meat in Romania.

The current study was conducted in order to analyze the prevalence of Escherichia coli (E. coli) in samples of chicken meat (100 chicken meat samples), as well as to evaluate the antimicrobial susceptibility of these isolates. A total of 30 samples were positive for E. coli among the collected chicken samples. Most isolates proved to be highly resistant to tetracycline (80%), ampicillin (80%), sulfamethoxazole (73.33%), chloramphenicol (70%) and nalidixic acid (60%). Strong resistance to ciprofloxacin (56.66%), trimethoprim (50%), cefotaxime (46.66%), ceftazidime (43.33%) and gentamicin (40%) was also observed. Notably, one E. coli strain also proved to be resistant to colistin. The antimicrobial resistance determinants detected among the E. coli isolates recovered in our study were consistent with their resistance phenotypes. Most of the isolates harbored the tetA (53.33%), tetB (46.66%), blaTEM (36.66%) and sul1 (26.66%) genes, but also aadA1 (23.33%), blaCTX (16.66%), blaOXA (16.66%), qnrA (16.66%) and aac (10%). In conclusion, to the best of our knowledge, this is among the first studies analyzing the prevalence and antimicrobial resistance of E. coli strains isolated from chicken meat in Romania and probably the first study reporting colistin resistance in E. coli isolates recovered from food sources in our country.

Prevalence and Characterisation of Clostridium perfringens Isolates in Food-Producing Animals in Romania.

The aim of the current study was to investigate the prevalence of Clostridium perfringens (C. perfringens) recovered from animal faeces, as well as to determine the antimicrobial susceptibility of such isolates. A total of 14 (14/100; 14%) C. perfringens isolates were isolated from the 100 analysed samples (twelve recovered from faecal samples collected from pigs and two from veal calves' faecal samples). The preponderant genotype was type A, with all isolates being cpa-positive. The most potent antimicrobial agents against C. perfringens proved to be vancomycin, rifampicin and lincomycin. A strong resistance to tetracycline (71.4%), penicillin (64.2%), erythromycin (42.8%) and enrofloxacin (35.7%) was also observed. To the best of our knowledge, this is the first analysis regarding the prevalence, characterization and antimicrobial susceptibility of C. perfringens in food-producing animals in Romania, adding further evidence for the probable role of animals as a source of resistant C. perfringens strains.

The Role of Wild Boars in the Circulation of Tick-Borne Pathogens: The First Evidence of Rickettsia monacensis Presence.

Most wild mammals can serve as hosts both for tick-borne pathogens (TBPs) and for the ticks themselves. Among these, wild boars, due to their large body size, habitat and life span, show high exposure to ticks and TBPs. These species are now one of the widest-ranging mammals in the world, as well as the most widespread suid. Despite the fact that certain local populations have been decimated by African swine fever (ASF), wild boars are still considered overabundant in most parts of the world, including Europe. Altogether, their long-life expectancy, large home ranges including migration, feeding and social behaviors, wide distribution, overabundance and increased chances of interactions with livestock or humans make them suitable sentinel species for general health threats, such as antimicrobial-resistant microorganisms, pollution and ASF geographical distribution, as well as for the distribution and abundance of hard ticks and also for certain TBPs, such as Anaplasma phagocytophilum. The aim of this study was to evaluate the presence of rickettsial agents in wild boars from two counties in Romania. Among 203 blood samples of wild boars (Sus scrofa ssp. attila) collected during 3 (2019-2022) hunting seasons (September-February), 15 were found positive for tick-borne pathogen DNA. Six wild boars were positive for A. phagocytophilum DNA presence and nine for Rickettsia spp. The identified rickettsial species were R. monacensis (six) and R. helvetica (three). No animal was positive either for Borrelia spp., Ehrlichia spp. or Babesia spp. To the best of our knowledge, this is the first report of R. monacensis in European wild boars, thus adding the third species from the SFG Rickettsia, in the epidemiology of which this wild species may have a role as a reservoir host.

Diversity of Thelazia spp. in domestic cattle from Romania: epidemiology and molecular diagnosis by a novel multiplex PCR.

BACKGROUND: Thelaziosis is a neglected vector-borne disease caused by parasitic nematode worms of the genus Thelazia which affects various hosts. Limited attention has been given to ungulate-associated Thelazia species. Current diagnosis of thelaziosis and the identification/differentiation of species heavily relies on morphological features. Therefore, we conducted an epidemiological study in Romanian cattle, with the aim to obtain morphological and molecular data that can be used for species identification. METHODS: The eyes of 705 slaughtered cattle were sampled and subjected to morphological identification, morphometric analysis, and molecular characterization. PCR amplification and sequence analysis were performed based on the cytochromec oxidase subunit 1 (COI) gene. Statistical tests assessed the correlations between infection parameters and ecological or biogeographical factors. A novel PCR method was developed based on the consensus sequence from each species. Specific forward primers were designed for each of the three species, and a reverse primer (COIintR) was used for all reactions. A consensus thermal profile was established by gradient PCR amplification of each species separately. RESULTS: Of the sampled cattle, 19.3% were infected with Thelazia spp. Prevalence varied significantly with ecogeographical factors. A total of 585 Thelazia nematodes were recovered, with T. rhodesi being the most abundant, followed by T. skrjabini and T. gulosa. Morphometric and molecular analyses supported the morphological identification, yielding unique sequences for each species. From the 59 T. rhodesi specimens sequenced, 29 unique sequences were obtained, with a 99.1-99.85% nucleotide identity to the only other COI sequence present in GenBank®. All nine T. gulosa isolates were unique (99.37-100% nucleotide identity to other sequences), while T. skrjabini specimens displayed 98.47-100% nucleotide identity to the sole available sequence. CONCLUSIONS: Bovine thelaziosis is prevalent in Romania, raising concerns for animal welfare and potential economic impacts. Infected cattle grazing alongside vulnerable wild ruminants, such as the European bison, may affect conservation efforts. Our newly developed multiplex PCR shows promise as a valuable surveillance tool, enabling the detection of occult infections in apparently healthy animals through lachrymal secretion testing.

Clostridioides difficile in Food-Producing Animals in Romania: First Study on the Prevalence and Antimicrobial Resistance.

At present, the epidemiology of the gastrointestinal disease caused by Clostridioides difficile (C. difficile) is starting to be slowly elucidated internationally, although information about the bacteria in the food supply chain is insufficient and, in many countries, even absent. The study was conducted in order to investigate the prevalence of C. difficile isolated from animal feces, as well as to determine the antimicrobial susceptibility of such isolates. The presence of antibiotic resistance determinants has also been evaluated. Overall, a total of 24 (12.5%) C. difficile isolates were recovered (out of the 192 samples collected), the highest percentage of positive isolates being detected in the fecal samples collected from piglets (25%). The majority of the isolates recovered in the current study proved to be toxigenic. Moreover, all C. difficile isolates were susceptible to vancomycin, although a large proportion of the porcine isolates (50%) were resistant to levofloxacin. The tetW and erm(B) genes have also been identified in the porcine isolates. In conclusion, this is the first analysis of the prevalence of C. difficile in food-producing animals in Romania, and it adds further evidence about the possible role of animals as a source of resistant C. difficile strains and a reservoir of antimicrobial resistance determinants.

Concerning Increase in Antimicrobial Resistance Patterns of Pathogenic Strains of Salmonella Isolated in Poultry Meat Products.

Salmonella is considered to be one of the major foodborne pathogens associated with the consumption of contaminated poultry meat products. To the best of our knowledge this is the first extended research performed on a number of Salmonella strains isolated during 2011-2021 from poultry meat products in Romania. The aim of this study was to characterize the prevalence of pathogenic Salmonella serovars, antimicrobial susceptibility, and antimicrobial resistance genes in 112 Salmonella isolates recovered from raw poultry meat products. The results showed that Salmonella enterica serovars Enteritidis and Typhimurium were the common serotypes (56%; 25%). Overall, the majority of the isolates were resistant to at least three tested antimicrobials. High resistance was observed for tetracycline (84%), nalidixic acid (78%), and ampicillin (78%) in pathogenic Salmonella isolated during the period 2016-2021. All the pathogenic Salmonella isolated during 2016-2021 tested positive to at least one resistance gene encoding for tetracycline resistance, with the tetA gene being the most prevalent (62%). In addition, 64% (24/37) of the Salmonella isolates carried at least one of the genes (blaCMY-2, blaSHV1, blaTEM1) that code for ß-Lactams resistance. The findings in this study showed a high prevalence of multi-drug resistant (MDR) Salmonella serovars in poultry meat products and a concerning increase of resistance patterns. The continuous occurrence of more resistant strains implies that effective measures should be strictly applied in this particular food chain in order to prevent their spread and guarantee microbial safety.

Pathogenic E. coli from Cattle as a Reservoir of Resistance Genes to Various Groups of Antibiotics.

Antimicrobial resistance has become a worldwide concern in all public health domains and reducing the spread has become a global priority. Pathogenic E. coli is responsible for a number of illnesses in humans and outbreaks in the past have been correlated with the consumption of contaminated bovine products. This is why surveillance in all the steps of production is essential. This study focused on identifying the pathogenic strains of E. coli in two large bovine abattoirs from Romania and France, and on associating them with the antimicrobial resistance patterns. A total of 250 samples from intestinal content were aseptically collected during the evisceration step of the cattle slaughtering process, from which 242 E. coli strains were isolated. Seventeen percent of all samples tested positive to at least one E. coli isolate carrying eaeA, stx1 and stx2 genes. The most prevalent genetic profile found in the E. coli strains tested was Stx1-positive and Stx2/eaeA-negative. More than 68% of the pathogenic E. coli isolated in Romania showed multi-drug resistance (MDR) and in France, the percentage was significantly lower (38%). The MDR profiles showed a high gene diversity for antibiotic resistance, which represents a great risk for environmental spread and human health. Our results indicate that in Romania, bovines can represent a reservoir for MDR E. coli and, hence, a surveillance system for antimicrobials usage in farm animals is highly needed.

Genotyping of African Swine Fever Virus (ASFV) Isolates in Romania with the First Report of Genotype II in Symptomatic Pigs.

The World Organisation for Animal Health has listed African swine fever as the most important deadly disease in domestic swine around the world. The virus was recently brought from South-East Africa to Georgia in 2007, and it has since expanded to Russia, Eastern Europe, China, and Southeast Asia, having a devastating impact on the global swine industry and economy. In this study, we report for the first time the molecular characterization of nine African swine fever virus (ASFV) isolates obtained from domestic pigs in Mures County, Romania. All nine Romanian samples clustered within p72 genotype II and showed 100% identity with all compared isolates from Georgia, Armenia, Russia, Azerbaijan, Ukraine, Belarus, Lithuania, and Poland. This is the first report of ASFV genotype II in the country.

Silver Nanoparticles for the Therapy of Tuberculosis.

Rapid emergence of aggressive, multidrug-resistant Mycobacteria strain represents the main cause of the current antimycobacterial-drug crisis and status of tuberculosis (TB) as a major global health problem. The relatively low-output of newly approved antibiotics contributes to the current orientation of research towards alternative antibacterial molecules such as advanced materials. Nanotechnology and nanoparticle research offers several exciting new-concepts and strategies which may prove to be valuable tools in improving the TB therapy. A new paradigm in antituberculous therapy using silver nanoparticles has the potential to overcome the medical limitations imposed in TB treatment by the drug resistance which is commonly reported for most of the current organic antibiotics. There is no doubt that AgNPs are promising future therapeutics for the medication of mycobacterial-induced diseases but the viability of this complementary strategy depends on overcoming several critical therapeutic issues as, poor delivery, variable intramacrophagic antimycobacterial efficiency, and residual toxicity. In this paper, we provide an overview of the pathology of mycobacterial-induced diseases, andhighlight the advantages and limitations of silver nanoparticles (AgNPs) in TB treatment.

Geographical origin and species differentiation of Transylvanian cheese. Comparative study of isotopic and elemental profiling vs. DNA results.

The present study proposed the cheese differentiation, according to geographical production area and with respect to species (cow, sheep) of two traditional cheese specialties, (salty and ripened), produced in Transylvania, Romania. For this purpose, the elemental profile and carbon isotopic ratios (13C/12C) of cheese and extracted casein were corroborated through statistic supervised techniques to get the best discrimination markers. The manganese content, along with Rare Earth Elements (REE) concentrations, proved to be very powerful predictors, for the traditional salted cheese mainly, due to the direct influence of the local salted water. Despite that proposed techniques are not acknowledged methods for species differentiation, this approach allowed a successful discrimination of the animal species that produced the raw materials for cheese manufacturing (milk). The results generated by the developed chemometric model, for species differentiation, were compared with those obtained using Isoelectric focusing (IEF) and DNA tests. The proposed association of isotopic and elemental markers allowed a differentiation better than 92% for geographical provenance, of each investigated cheese specialties while, for species discrimination (cow vs. sheep) a percentage of 100% was obtained.

First study on characterization of virulence and antibiotic resistance genes in verotoxigenic and enterotoxigenic E. coli isolated from raw milk and unpasteurized traditional cheeses in Romania.

The study focused on the incidence of enterotoxigenic Escherichia coli (ETEC) and verotoxigenic E. coli (VTEC) in raw milk and traditional dairy cheeses marketed in Romania, characterizing the virulence and antibiotic resistance genes of these isolates. One hundred and twenty samples of raw milk and 80 samples of unpasteurized telemy cheese were collected and cultured according to the international standard protocol. All the characteristic E. coli cultures were analyzed for the presence of STa, STb, LT, stx1, and stx2 toxicity genes. The ETEC/VTEC strains were tested for the presence of antibiotic resistance genes, such as aadA1, tetA, tetB, tetC, tetG, dfrA1, qnrA, aaC, sul1, bla SHV , bla CMY , bla TEM , and ere(A), using PCR. The results showed that 27 samples (18.62%) were positive for one of the virulence genes investigated. 48.1% (n = 13) tested positive at the genes encoding for tetracycline resistance, tetA being the most prevalent one (61.5%; n = 8). A high percent (33.3%; n = 9) revealed the beta-lactamase (bla TEM ) resistance gene, and none of the samples tested positive for bla CMY and bla SHV genes. The genes responsible for resistance to sulfonamides (sul1) and trimethoprim (dfrA1) were detected in rates of 14.8% (n = 4) and 7.4% (n = 2), respectively. E. coli is highly prevalent in raw milk and unpasteurized cheeses marketed in Romania. These strains might represent an important reservoir of resistance genes which can easily spread into other European countries, given the unique market.

Concerning Increase in Antimicrobial Resistance in Shiga Toxin-Producing Escherichia coli Isolated from Young Animals during 1980-2016.

This study was conducted in order to assess the antimicrobial resistance patterns of E. coli isolated from young animals affected between 1980 and 2016. The selected isolates for this study (n=175) carried stx1/stx2 genes and the most prevalent type of pathogenic E. coli found belonged to serogroup O101, antigen (K99)-F41 positive. All STEC-positive isolates were tested for susceptibility to 11 antimicrobials. Multidrug resistance (MDR) increased from 11% during the 1980s to 40% between 2000 and 2016. Resistance to tetracycline and streptomycin was the most frequent co-resistance phenotype (37%). Co-resistance to tetracycline and sulfonamide was found in 21% of E. coli isolates, while the MDR pattern to tetracycline, sulfonamide, and streptomycin was observed in 12% of the strains tested. Only 8% of isolates were co-resistant to tetracycline, ampicillin, streptomycin, and sulfonamide. The most common resistance genes found were those encoding for tetracycline, sulphonamides, and streptomycin, with 54% (n=95) of the tested isolates containing at least one of the genes encoding tetracycline resistance. A total of 87% of E. coli that tested positive for tetracycline (tetA, tetB, and tetC) and sulphonamide (sul1) resistance genes were isolated between 2000 and 2016. A large number of isolates (n=21) carried int1 and a nucleotide sequence analysis revealed that all class 1 integron gene cassettes carried sul1, tet, and dfrA1 resistance genes. An increase was observed in the level of resistance to antimicrobials in Romania, highlighting the urgent need for a surveillance and prevention system for antimicrobial resistance in livestock in Eastern Europe.

Rabies surveillance in the rural population of Cluj County, Romania.

INTRODUCTION: The rabies virus causes an acute encephalomyelitis that progresses to coma and death within 10 days of the onset of the disease. Taking into account the fatality of the disease and the absence of a specific treatment, preventive methods are of the outmost importance. The purpose of this study was to assess the exposure of the rural population from Cluj County, Romania, to rabies risk factors. METHODS: Data regarding the exposure of the rural population from Cluj County to rabies risk factors were collected from the Vaccination Center registry at the Infectious Diseases Teaching Hospital in Cluj-Napoca. The information was treated confidentially. The data obtained were collected using an Access 2000 database and was analyzed using Epi Info. Information regarding the general population was available from the 2002 General Population and Housing Census. RESULTS: Cluj County reported 1008 cases of human exposure to potentially rabid animals between April 1998 and December 2004. The relative contributions of the major groups of aggressor animals were as follows: 882 dogs (81.55%), 51 cats (5.06%), 30 foxes (2.98%), 28 horses (2.78%), 19 rats (1.89%), and 58 other animals (5.74%). Post-exposure prophylaxis with antirabies serum immune globulin was prescribed in 9% of the cases (n = 90). Vaccine was recommended in 72% of the cases (n = 726). Two hundred and eighty-three patients out of 726 (38.98%) underwent complete vaccine prophylaxis. Diagnoses in animals suspected of having rabies were made by direct immunofluorescent antibody staining of rabies viral antigen in brain material. The number of animals that tested positive for rabies during the period April 1998 to December 2004 was 49. More than 77% (n = 38) were wild animals. The relative contributions of the major groups of animals were as follows: 33 foxes (67.35%), 7 dogs (14.29%), 2 wolves (4.08%) and 7 other animals (14.29%). CONCLUSIONS: The fatal risk of rabies in human beings, and the persistence of the virus in this geographic area, makes legitimate the recommendation of prophylactic procedures for persons exposed to potentially rabid animals.

Antibiotic susceptibility and prevalence of foodborne pathogens in poultry meat in Romania.

INTRODUCTION: The occurrence of pathogenic strains in poultry meat is of growing concern in Romania. Another problem found on a global level is the continuous increase of antimicrobial resistance in bacteria isolated from food. This study aimed to evaluate the prevalence of pathogenic bacteria in poultry carcasses obtained in Romania in 2012-2013 and to reveal the most prevalent patterns of antimicrobial resistance in the isolated strains. METHODOLOGY: A total of 144 broiler chicken carcasses were evaluated according to classical microbiological methods. The DNA was extracted from the bacterial colonies and the resistance genes were identified by PCR. RESULTS: In 2012, 47.2% of the samples revealed at least one of the following bacteria: Campylobacter jejuni (9.72%; n = 7), Salmonella enterica serotype Enteritidis (4.17%; n = 3), Listeria monocytogenes (15.28%; n = 11), and Escherichia coli (16.67%; n = 12). In 2013, the number of positive samples of pathogenic bacteria decreased, although Campylobacter jejuni was isolated in a higher percentage (20.8% vs. 9.72%). The percentage of multidrug-resistant (MDR) bacteria was high (23%); the most prevalent pattern included resistance to tetracycline, sulfonamides, and quinolones/fluoroquinolones. All the resistant Salmonella and E. coli strains were tested for the presence of characteristic resistance genes (Kn, bla(TEM), tetA, tetB, tetG, DfrIa, aadA1a, Sul) and revealed that these isolates represent an important reservoir in the spread of this phenomenon. CONCLUSIONS: Our findings suggest that Romania urgently needs an integrated surveillance system within the entire chain, for drug-resistant pathogens isolated from poultry meat.

First description of plasmid-mediated quinolone resistance determinants and ß-lactamase encoding genes in non-typhoidal Salmonella isolated from humans, one companion animal and food in Romania.

BACKGROUND: Gastroenteritis attributable to Salmonella enterica and the continuous increase in antimicrobial resistance of this gut pathogen, which compromises the use of previously effective treatments, is of great concern for public health. This study was conducted in order to investigate the presence of plasmid-mediated quinolone resistance (PMQR) determinants and ß-lactamase-encoding genes, in S. enterica, isolated from humans, one companion animal and food. Moreover, the study aimed to identify potential vehicles of transmission of resistant strains to humans, with focus on food products (meat). METHODS: A total of 20 S. enterica isolates recovered from food (chicken and pork meat), one companion animal and humans (stool samples), were examined for their serotype, antimicrobial susceptibility and the presence of PMQR and ß-lactamase-encoding genes. Moreover, the genetic relatedness of nine Salmonella Infantis and ten Salmonella Enteritidis isolates was analyzed by pulsed-field gel electrophoresis (PFGE). RESULTS: Among all isolates, 15 (75%) were multidrug-resistant (MDR) and the majority of them proved to be resistant to nalidixic acid and fluoroquinolones (FQs) (ciprofloxacin and levofloxacin). Twelve isolates (60%) harboured at least one PMQR gene [qnrA, qnrB, qnrS, aac (6')-Ib-cr or qepA] while seven isolates (35%) carried at least one ß-lactamase-encoding gene (bla TEM, bla PSE-1, bla SHV or bla CTX-M). Moreover, two or more PMQR or ß-lactamase-encoding genes co-existed in a single S. enterica isolate. A number of nine Salmonella Infantis, as well as the majority of Salmonella Enteritidis isolates analyzed by PFGE proved to be closely related. CONCLUSIONS: The study demonstrated the co-existence of PMQR and ß-lactamase-encoding genes among the Salmonella isolates recovered and confirmed that multiple mechanisms might be involved in the acquisition and spread of resistance determinants. The close genetic relatedness between the clinical and foodborne S. enterica isolates, suggested that chicken meat might be a possible cause of human salmonellosis in our country, during the study period. Results of this study might improve understanding of the antimicrobial resistance mechanisms and transmission dynamics of Salmonella spp. Here, we report for the first time the presence of PMQR and ß-lactamase-encoding genes in S. enterica isolates, recovered from humans, one companion animal and food, in Romania.

First study of Salmonella in meat in Romania.

INTRODUCTION: The increasing antimicrobial resistance of Salmonella isolates is of major public health concern, but information regarding these aspects is still lacking in Romania. This study focused on a detailed and accurate investigation concerning prevalence, serotypes, and antimicrobial resistance patterns of Salmonella strains, isolated from pork and chicken meat, collected from all regions of Romania in 2011. METHODOLOGY: The research was conducted on 650 samples of chicken and pork meat collected from production units and retail markets located in various regions of Romania. A total of 149 Salmonella isolates were recovered (22.92%), serotyped, confirmed by PCR, and further tested for antimicrobial susceptibility. RESULTS: Thirteen Salmonella serovars were identified; predominant serovars included Infantis, Typhimurium, Derby and Colindale. Multiple resistance was found in 83.22% (n = 124) of the isolates. The isolates were frequently resistant to tetracycline (80.53%), streptomycin (81.21%), sulfamethoxazole (87.25%), nalidixic acid (65.10%), and ciprofloxacin (42.95%). Additionally, a markedly lower resistance rate was observed for ampicillin (20.81%), chloramphenicol (16.78%), and ceftazidime (11.41%). Among 137 resistant Salmonella isolates, 35 different resistance patterns were found. CONCLUSION: A high prevalence of Salmonella spp. and a relatively high resistance rate to multiple antimicrobials was found. This data indicates that chicken and pork meat could constitute a source of human exposure to multidrug-resistant Salmonella and therefore could be considered a potential vehicle of resistant Salmonella foodborne diseases. Further actions are needed to succesfully implement a national surveillance program for better monitoring of these resistant pathogens.