Virological supervision of bluetongue disease in the south-east region of Romania

Bluetongue is an infectious, non-contagious, vector-transmitted viral disease affecting domestic ruminants (sheep, goats, cattle) and wild (buffaloes, deer, several species of African antelopes and other species of the Artiodactyla order). The economic importance of the disease lies in the important economic losses following the decrease in the productive capacity of the animals, mortality and fetal malformations, immunization costs of the receptive animals, trade restrictions, reduction of the economic recovery price of the receptive animals and products thereof. Our study aimed at identifying by virological examinations the presence and circulation of BLA virus in the SE region of Romania. For viral isolation and identification were used blood samples collected from domestic ruminants in the counties: Galati, Braila, Tulcea and Vrancea. According to the working chart of the BT Diagnostic Manual (LNR Arboviroze Bucharest), the samples collected from suspect animals were processed and tested by RT-PCR. In the period 2015-2016, 517 blood samples with anticoagulant from 282 cattle and 235 sheep suspected of Bluetongue were tested for the identification of the viral genome by RT-PCR technique. There were no suspicions of Bluetongue disease in goats in the counties included in the study. In bovines in the SE of Romania, the viral genome was identified in 171 (60.64%) blood samples with anticoagulant. In sheep in the SE of Romania, the viral genome was identified in 209 (88.93%) blood samples with anticoagulant. Most positive samples confirmed by the detection of the BT viral genome came from Vrancea, both in cattle (161 positive samples) and in sheep (209 positive samples). Because of the pathogenicity, bluetongue virus infection can not be diagnosed for a certain period of time, the period in which the disease may exist and evolve, the infected animals being sources of infection for vectoric culicoid insects

Characterisation of Extended β-Lactamases and Plasmid Mediated Quinolones Resistancein Escherichia Coli from Shelter Dogs

The aim of this study was to determine the prevalence of β-lactamase (TEM, SHV, OXA), extended-spectrum β-lactamase (ESBL) and genes encoding plasmid mediated resistance to quinolones (PMQR) in extended spectrum cephalosporin (ESC)-resistant Escherichia coli isolated from dog faeces from two shelters in the North-East of Romania. Eighty-eight faecal samples from healthy dogs were analysed by cultivation on Brilliance ESBL medium (Oxoid, UK), followed by phenotipic ESBL screening using combination disc test (CDT). Identification of the E. coli strains was performed by uidA/uspA gene PCR. Susceptibility testing was performed on Mueller-Hinton Agar, with β-lactam and non-β-lactam agents. Identification of β-lactamase genes (blaCTX-M, blaTEM, blaSHV, blaOXA) and PMQR genes (qnrA, qnrB and qnrS) was performed by PCR as previously described. Twenty eight ESC-resistant E. coli (31.81%) were obtained and (n=21/28, 75%) of these were confirmed as ESBLs and showed resistance to cefpodoxime (n=21/28, 75%), amoxicillin/clavulanic acid (n=19/21; 90.48%), and enrofloxacin (n=8/21; 38.09%). Predominant ESBL types were CTX-M-1 (n=15/17, 88.24%) and CTX-M-9 (n=2/17, 11.76%) enzymes. TEM and SHV enzymes were identified in 17.86% and 14.29% of the ESC-resistant isolates, whilst some isolates (n=4) carried only blaTEM and blaSHV. The prevalence of PMQR genes was 28.57% of the 28 ESC resistant isolates, consisting of qnrS (62.5%) and qnrB (37.5%). These findings indicate a high prevalence of ESBLs and PMQR associated resistance E. coli in the normal faecal microbiota of dogs from shelters, which carries the risk for dissemination of these resistance genes to other animals, human or the environment

Comparative evaluation of three testing methods for detection of mediated resistance MBL in pseudomonas aeruginosa isolates

Metallo – β – lactamase (MBL) producing Pseudomonas aeruginosa have been reported to be important cause of nosocomial infections. The appearance of MBL genes and their spread among bacterial pathogens is a matter of concern with regard to the future of antimicrobial therapy. The present study was undertaken to determine which method is better to use in laboratory for detecting MBL producing P. aeruginosa. A total of 182isolates of P. aeruginosa from human and animals, 125 from human and 57 from animals, (burns, pus, urine, blood cultures, etc.), collected between 2013 and 2015 were subjected to susceptibility testing against various antibiotics by disc diffusion test according to Clinical and Laboratory Standards Institute (CLSI) guidelines 2015. Imipenem resistant isolates were selected for the detection of MBL production by E-test strips for screening MBL, double disc method (IPM and IPM+EDTA) and EDTA solution application on microcaps IPM. The positive results have been based on inhibition zone around imipenem discs impregnated with EDTA as compared to those without EDTA confirmed MBL production and for E-test strips the strains were positive those that have developed around area with EDTA solution. The double disc method (IPM and IPM+EDTA is most effective way to use in laboratory to determine early producer P. aeruginosa MBL, having 2 advantage: first is the low cost for materials (MH agar and microcaps) and the technique is very easy to be applied

Assessment of antibiotics sensitivity of the most frequent potential pathogen bacteria isolated in the microbiology laboratory of the Faculty of Veterinary Medicine Iași, during 2017-2018

In the veterinary practice, establishing the etiological diagnosis represents the quintessence of the anti- infectious therapy. The final diagnosis based on the correctness of the laboratory results and the microbiological examination, together with the hematological, immunological, histopathological, etc., has a major role in getting the right therapeutic protocol. In the current context, characterized by a wide etiological variety of infections, it is necessary to identify and test the sensitivity to antibiotics of pathogens isolated from different biological samples. The present study of the Microbiology Laboratory of the Faculty of Veterinary Medicine Iași presents the results obtained on various strains isolated from dogs and cats with different diseases during 2017-2018. The tests performed on of 83 microbiological samples (otic, pharyngeal, cutaneous and conjunctival secretions, urine, feces, etc.) identified 107 aerobic and anaerobic bacterial strains, classified into 20 bacterial genera. The most commonly isolated aerobic bacterial species were: Staphylococcus pseudointermedius (27.10%), Streptococcus sp.gr.G (8.41), Enterococcus faecalis (4.67%), Streptococcus sp. gr.C (3.73%), Pseudomonas aeruginosa (5.60%), Klebsiella pneumoniae (2.80%). The most commonly isolated anaerobic bacterial species were: Clostridium perfringens (9.35%), Campylobacter sp. (1.87%).The results of the antibiograms revealed a wide variability of sensitivity and resistance of the isolated strains to the antibiotics, most of them being multiple drug resistance

Clostridium perfringens enterotoxigen involved in hemorrhagic diarrhea at dogs

Clostridium perfringens is a commensal of the human and animal intestines. The toxigenic strains of this bacterial species are responsible for some enteral diseases in humans and domestic animals. In dogs, hemorrhagic enteritis produced by Clostridium perfringens are sporadic but have a severe progression-restricted progression. In the year 2018, 9 faeces samples were taken from dewormed dogs, vaccinated but with enteritis and associated toxic conditions (vomiting, lethargy, diminished appetite) that started suddenly without any other clinical history. Based on the anamnesis, it excluded the risk of chemical poisoning. Faeces samples were subjected to the microbiological exam. The bacterioscopic examination in Gram stained smears was predominantly Gram positive bacilli with morphology and characteristic disposition for Clostridium sp and no specific morphological phenotypes for spirochetes or protozoa were identified. The samples were incubated at 37° C, under anaerobic conditions, on liquid culture media (bullion VL) and solids (Clostridium agar, SPS agar) and under aerobic conditions on culture media for aerobic bacteria (nutrient broth, blood agar) (E. coli, Salmonella sp., Shigella sp., Campylobacter sp., Yersinia sp., Serpulina sp., Vibrio sp., etc.) which can trigger enteritis in dogs. Following the bacteriological examination, strains of Clostridium sp. Biochemical tests have included the species as Clostridium perfrigens. The clinical progression in the 9 patients was different: 6 dogs responded to antibiotic therapy, recommended on the basis of the antibiotic, and 3 dogs died within 48 hours before a treatment was instituted

Necrotic enteritis in meat chicken raised at the ground in permanent bedding

Poultry necrotic enteritis is an acute clostridial infection characterized by severe necroses of intestinal mucosa. The disease begins suddenly, with a sharp increase in death rate and dehydration. Clostridium perfringens, a sporulated, anaerobic, Gram-positive bacterium is commonly found in the environment and in the gastrointestinal tract as part of the normal intestinal flora. Frequent presence in the digestive tract of healthy birds is associated with necrotic enteritis in broilers. The research was conducted on 323 samples (120 live chickens, 89 corpses, 104 feed samples and 10 water samples) collected from a farm with 32 253 hybrid Ross 308 broilers (21 days), raised at the ground on permanent bedding, where there was a significant increase in mortality above the permissible limit. The necropsy performed on 980 chicken corps revealed a different prevalence of intestinal tract lesions: bleeding wall (28.37%), mucosal necrosis (23.22%), gas content (18.57%), mucosal inflammation (15.73%) and red orange mucus in the intestines (14.10%). Bacteriological examination identified Clostridium perfringens in 11.66% of live broilers, 10.11% of chicken corps, 61.53% of feed samples and 3.09% of water samples. Increased percentage this species isolation suggests that feed taken from the hall was an important source of infection for broilers reared on the ground

Investigations regarding on the clinical expression of Bluetongue virus infection of domestic ruminants in the South-East region of Romania

Bluethonge has a significant economic impact, mainly due to the effect of the disease on animals (morbidity, mortality, reproductive insufficiency, reducced of milk production and animal weakening) and, in particular, disruption of international trade in animals and animal products. In the South-East region of Romania, in the period 2014-2017, animals with clinical signs specific to Bluetongue virus were identified.1437 domestic ruminants with a suspected disease were examined. Of these, 418 (29,08%) had clinical features similar to those of Bluetongue. The incidence of clinical signs in animals with suspected disease varied by species. Thus, the clinical aspects were present in 22,21% cattle, 68,21% sheep and 1,54% goats. Following clinical examination, the simultaneous presence of several clinical signs specific for Bluetongue virus infection was observed in cattle and sheep. In cattle, the incidence of clinical signs was variable, most frequently reported conjunctival mucosal hyperemia, epiphora, sialorrhea, gingival ulceration, swelling and cyanosis of the tongue, nasal edema with ulcerations of the nasal mucosa and muco-purulent jet, circumcised necrosis at the level of the udder, accentuated weakening, congestion of the coronary area, torticollis, digestive disorders, exongulations and inability to move. The pathogenesis of Bluetongue is similar in cattle and sheep, so that the clinical evolution was similar in the two species. In goats, the disease evolved inapparently or with diminished clinical expressions: prostate condition, mucopurulent discharge, edema and nasal ulcers, cyanosis of the mucous membranes, curls and limping. In addition to the immediate losses, the onset of the disease can generate long-term effects, which can decisively affect the Romanian animal breeding sector. The ease with which the Bluetongue disease has spread, shows that the territory of Romania is affected by climate change caused by global warming, which allowed the transmitting vectors to proliferate and spread the Bluetongue virus to the receptive animals