The present study aimed to determine the bacteria isolated from bovine mastitis and their antimicrobial resistance in the western part of Romania. Clinical mastitis was diagnosed based on local inflammation in the udder, changes in milk, and when present, generalized symptoms. Subclinical mastitis was assessed using a rapid test—the California Mastitis Test. The identification of bacterial strains was performed based on biochemical profiles using API system tests (API 20 E, API Staph, API 20 Strep, API Coryne, API 20 NE (bioMerieux, Marcy l’Etoile, France), and MALDI-TOF mass spectrometry (MS). The prevalent isolated bacteria were Staphylococcus spp. (50/116; 43.19%), followed by Streptococcus spp. (26/116; 22.41%), E. coli (16/116; 13.79%), Corynebacterium spp. (9/116; 7.75%), Enterococcus spp. (10/116; 8.62%), and Enterobacter spp. (5/116; 4.31%). Phenotype antimicrobial resistance profiling was performed used the disc diffusion method. Generally, Gram-positive bacteria showed low susceptibility to most of the antimicrobials tested, except cephalothin. Susceptibilities to penicillins and quinolones were fairly high in Gram-negative bacteria, whereas resistance was observed to macrolides, aminoglycosides, and tetracyclines. The highest number of isolates were multidrug resistant (MDR), the resistance pathotypes identified including the most frequently antimicrobials used in cow mastitis treatment in Romania.
This study aimed to investigate the antioxidant profile and the antimicrobial activity of four different types of monofloral honey (manuka (MH), brassica rapeseed (BH), acacia (AH), and linden honey (LH)) against some bacterial/fungal ATCC strains and some multidrug-resistant strains isolated from chronic otitis in dogs. For the characterisation of the antioxidant profile of each honey, we extracted the honey samples by hydroalcoholic extraction and analysed them in terms of total polyphenols (TPC), total flavonoids (TFC), and 2,2-diphenyl-1-picrylhydrazyl (DPPH) using the spectrophotometric method. The antimicrobial activity was determined using the microdilution method at concentrations of 10%, 15%, and 20%, with the results expressed in OD (optical density) calculated as BIR% (bacterial inhibition rate)/MIR% (mycelial inhibition rate). The antioxidant characterisation of the analysed honey samples showed the highest antioxidant activity and concentrations of TPC and TFC in MH, followed by LH. MH was proven to be the most effective on most clinical isolates concerning the antimicrobial activity in comparison with BH, AH, and LH. Except for B. cepacia and P. vulgaris, all the clinical isolates were sensitive to the antibacterial activity of honey. Regarding the ATCC strains, MH 10% was the most effective in inhibiting all the strains tested except for P. aeruginosa. In conclusion, the efficacy classification in our study was MH > BH > AH > LH.
Bacterial contamination of boar semen occurs with some frequency in artificial insemination centers and may have a negative effect on the quality of the semen as well as on the sows’ reproductive capacity. Normally, the source of bacterial contamination in pig seminal doses is the own boar. However, distilled water or laboratory equipment used to elaborate the seminal doses can be an important source of bacterial contamination. This study focused on the identification of gram-negative bacteria in boar semen, and impact on the quality of ejaculates obtained from boar, as well as on the establishment of antimicrobial resistance patterns of isolated gram-negative bacteria. Semen samples were collected from 96 boars, ranging in age from 12–36 month, from three artificial insemination centers from the North-West of Romania. Bacterial species were identified by two methods: matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry and API 20 E (BioMerieux, France). The main bacteria isolated from the doses diluted semen were gram-negative bacteria (47.91%), with a majority of the contaminant bacteria belonging to the family Enterobacteriaceae: Seratia marcescens 19.56%, Proteus mirabilis 15.21% and Escherichia coli 10.86% and to the family Pseudomonaceae: Ralstonia picketii 17.39%, Burkholderia cepacia 10.86%, Pseudomonas aeruginosa 8.69%, and Pseudomonas fluorescens 4.34%, respectively. More than half of these isolates (56.52%) were resistant to gentamycin and 58.69% were resistant to penicillin. These antibiotics are very frequently added in sperm diluent in the centers for the processing of sperm from boars in Romania. Regular monitoring for bacterial contamination is an important aspect of a control program.
The present study aimed to determine the prevalence of Staphylococcus species, which pose risks for public health, by evaluating skin samples collected from dogs in an animal shelter in Timisoara. Skin samples were taken from 78 dogs, which were either clinically healthy or suffering from dermatological conditions. Staphylococcus spp. was isolated and recognized based on conventional methods based on colony appearance, microscopic morphology, sugar fermentation, and coagulase activity. Following biochemical analysis, Staphylococcus isolates were subject to PCR tests to detect sa-f and sa-r genes to confirm the isolates to genus level. The typical colonies were identified to species level using biochemical methods, namely the VITEK®2 ID-GP64 identification card (bioMerieux, France). The phenotypic antimicrobial resistance profiling was performed using the VITEK®2 AST GP Gram-positive specific bacteria card (bioMerieux, France). Forty-three samples were confirmed as positive for Staphylococcus spp. Staphylococcus isolates were classified into the following categories: S. aureus, S. pseudintermedius, S. intermedius, S. epidermitis, S. haemolyticus, and S. hyicus. Eight (18.60%, 8/43) out of all the samples harbored the mecA gene, highlighting the distribution among isolated staphylococcal species: Staphylococcus pseudintermedius (4/43, 9.30%), Staphylococcus intermedius (1/43, 2.32%) and Staphylococcus aureus (3/43, 9.30%), respectively. The phenomenon of resistance was present, to the following antimicrobial agents: erythromycin (38/43, 88.37%), benzylpenicillin, kanamycin, and tetracycline with 37 strains (37/43, 86.04%), gentamycin (30/43, 69.76%), chloramphenicol (29/43, 67.44%), trimethoprim/sulfamethoxazole (27/43, 62.79%), ampicillin (26/43, 60,46%), rifampicin (25/43, 58,13%), imipenem (14/43, 32,55%), nitrofurantoin (11/43, 25.58%), oxacillin (8/43, 18.60%), vancomycin (4/43, 9.30%) and clindamycin (3/43, 6.97%), respectively. The presence of multidrug-resistant zoonotic staphylococci in clinically healthy dogs and dogs with skin lesions is an animal health and human health concern.
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