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

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

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

The influence of Ph and temperature on Salmonella spp. from fresh, chilled and frozen poultry carcasses

The trials were conducted in a slaughtering unit from Iaşi, where 192 samples from poultry carcasses were gathered and analysed microbiologically. Carcasses were washed in order to obtain test samples. The main sources of salmonellosis were poultry and poultry products. Salmonella is one of the most important worldwide causes of foodborne diseases. In order to reduce the contamination, we have treated experimentally the washing water with lactic acid sol. 1%. Thus, we stopped the evolution of the microorganisms susceptible to environmental pH changes. This pH change reduced the microbial load, especially of Salmonella spp,. from the surface of the carcass. The pH of the carcass washing water was 6.75; after the addition of lactic acid, it reached 2.34 and the temperature of washing water was 150C. The contamination of fresh carcasses with Salmonella spp had an incidence of 14.06%. After the treatment of washing water, it decreased until 4.6%. In case of 40C chilled carcasses, the incidence of Salmonella species in the presence of untreated carcasses was of 4.6%, being reduced to 1.5% after treatment

Dynamics of the susceptibility to different antibiotics and chemical-therapy treatments of isolated germs from puerperal cow genital secretions

The study was conducted on a group of 10 cows, belonging to Bălţată cu Negru Românească breed, at the Cattle Research and Development Station of Dancu , Iaşi district. Genital secretions were sampled from puerperal cows (first 4 post-partum weeks), for determining the uterine bacterial flora and germ susceptibility to different antimicrobial products. The obtained results have shown a lower germ susceptibility in the first post-partum week, compared to the next weeks. In weeks 2,3 and 4 post-partum , the mean values of germ susceptibility to antibiotics had very high levels (90-100%) to Rifampine, Erythromycin, Cephalotin, average levels (70-83.3%) to Amoxicillin, Kanamycyn, Chloramphenicol, Streptomycin, Gentamicyn, bacteria showing resistance to Penicillin, Oxacicllin, Sulphametazol, Methycillin. The most frequent bacterium isolated from genital secretions of cows with inflammatory genital troubles was Arcanobacterium pyogenes, in single bacterial colony (31.6%) or in mixed bacterial colony with E.coli (5.3%)

The first case of isolation and identification of acinetobacter radioresistens in infant from Iasi county (case report)

Members of the genus Acinetobacter are described as gram-negative, strictly aerobic diplococcoid rods that are oxidase negative and catalase positive. The genus includes at least 19 genomic species, defined on the basis of DNA relatedness criteria, which are ubiquitous in nature and have become increasingly responsible for a range of systemic infections in critically ill and immunocompromised patients. In most clinical microbiology laboratories, identification of Acinetobacter cannot routinely be achieved at the genospecies level because commercial identification systems are substantially deficient and poorly discriminatory in distinguishing these organisms. This implies that local data on the prevalence of individual species in human infections should be interpreted cautiously. In this article we describe a case study of community acquired Acinetobacter radioresistens. Bacteriae have been isolated from blood culture was isolate from one child, 9 month age, hospitalized in oncology sections, with the usual method of identification, the medium was seeded with blood cultures, it not developed in McK agar, but grows very well on blood agar at 37⁰C for 24 h. Biochemical confirmation was achieved using galleries API 20 NF. In conclusion, it is very important to identify Acinetobacter radioresistens accurately because of being a silent source of carbapenem resistance for Acinetobacter spp

Determination of biofilm production in animals originated pseudomonas aeruginosa strains

For the detection of Biofilm formation method, total 56 clinical isolates viz. Pseudomonas aeruginosa were used. Strains were identified using standards microbiological procedure. The susceptibility test of biofilm producing bacteria was performed by using disc diffusion technique. Biofilm detection was tested with Congo Red Agar method (CRA). These method is rapid, sensitive and reproducible, this method is suitable for detection of biofilm formation in the present study. Out of 56 isolates, CRA method detected 17 (30,35%) as high biofilm producer, and 39 (69,65 %) biofilm non- producer. According to the antibiotic susceptibility test, higher antibiotic resistance was observed in biofilm producing bacteria than non-biofilm producers