Antimicrobial resistance patterns and biofilm formation of coagulase-negative Staphylococcus species isolated from subclinical mastitis cow milk samples submitted to the Onderstepoort Milk Laboratory

BACKGROUND : Increased prevalence of antimicrobial resistance, treatment failure, and financial losses have been reported in dairy cows with coagulase-negative Staphylococcus (CoNS) clinical mastitis, however, studies on CoNS infections are limited in South Africa. Therefore, the objectives of this study were to investigate the antimicrobial resistance patterns and biofilm formation in CoNS isolated from cow milk samples submitted to the Onderstepoort Milk Laboratory. RESULTS : A total of 142 confirmed CoNS isolates were used for this study. Biofilm formation was identified in 18% of CoNS tested. Staphylococcus chromogenes (11%) had the highest proportion of biofilm formation followed by S. haemolyticus (4.0%), S. epidermidis, S. hominis, S. xylosus, and S. simulans with 1% respectively. Ninety percent (90%) of CoNS were resistant to at least one antimicrobial (AMR) and 51% were multidrug-resistant (MDR). Resistance among CoNS was the highest to ampicillin (90%) and penicillin (89%), few isolates resistant to cefoxitin and vancomycin, 9% respectively. Similarly, MDR-S. haemolyticus (44%), MDR-S. epidermidis (65%), and MDR-S. chromogenes (52%) were mainly resistant to penicillins. The most common resistance patterns observed were resistance to penicillin-ampicillin (16%) and penicillin-ampicillin-erythromycin (10%). Only 42% of biofilm positive CoNS were MDR. CONCLUSION : The majority of CoNS in this study were resistance to penicillins. In addition, most isolates were β- lactam resistant and MDR. Biofilm formation among the CoNS in this study was uncommon and there was no significant difference in the proportion of MDR-CoNS based on the ability to form a biofilm.The Research Development Programme (RDP) of the University of Pretoria and the Milk Producers’ Organisation (MPO).https://bmcvetres.biomedcentral.comam2020Paraclinical SciencesProduction Animal Studie

Antimicrobial resistance among Escherichia coli isolates from dogs presented with urinary tract infections at a veterinary teaching hospital in South Africa

Abstract Background This study investigated the burden and predictors of canine E. coli urinary tract infections (UTI) and antimicrobial resistance among dogs presented at a veterinary teaching hospital in South Africa, 2007–2012. Methods The Cochran-Armitage trend test was used to investigate temporal trends while logistic regression models were used to investigate predictors (age, sex, breed, year) of E. coli infections and antimicrobial resistance (AMR). Results A total of 22.3% (168/755) of the urinary specimens tested positive for E. coli. A significant (p = 0.0004) decreasing temporal trend in the percentage of E. coli positive isolates was observed over the study period. There were high levels of AMR to penicillin-G (99%), clindamycin (100%), tylosine (95%), cephalothin (84%) but relatively low levels of resistance to enrofloxacin (16%), orbifloxacin (21%). Almost all (98%, 164/167) the isolates exhibited multidrug resistance (MDR), while only 11% (19/167) and 2% (4/167) exhibited extensive drug resistance (XDR) and pan-drug resistance (PDR), respectively. Conclusions Although, the risk of E. coli UTI declined during the study period, the risk of AMR increased. The high levels of AMR and MDR as well as the presence of XDR and PDR is concerning as these have the potential of affecting prognosis of UTI treatments

Species identification and cow risks of non-aureus staphylococci from South African dairy herds

DATA AVAILABILITY : Data supporting the findings of this study are available from the corresponding author, I.-M.P., on request.Detailed information on specific species of non-aureus staphylococci (NAS) has become a necessity for effective udder health control programs in South Africa. The main objective of this preliminary study was to identify the different NAS species and strains present in dairy herds in South Africa using a cost-effective method. A further objective was to investigate the effects of cow risk factors and farming systems on the NAS isolates identified. A total of 214 NAS, isolated from milk collected from 17 South African dairy herds, were identified using three diagnostic tests (API Staph test, MALDI-TOF and 16s rRNA). There was a good observed agreement between the MALDI-TOF and 16S rRNA sequencing (92.2%) and a poor observed agreement between the MALDI-TOF and API Staph (25.7%). The genetic relatedness within species was investigated in 128 of these isolates using random polymorphic amplified deoxyribonucleic acid (DNA) (RAPD), verified by multilocus sequence typing (MLST), and phylogenetic analysis and cow risk factors were investigated on species level. The main NAS species isolated were Staphylococcus chromogenes (75.2%), Staphylococcus epidermidis (9.4%) and Staphylococcus haemolyticus (8.9%). The RAPD test identified 34 Staphylococcus chromogenes, 13 Staphylococcus epidermidis and nine Staphylococcus haemolyticus strains, indicating genetic diversity amongst strains and herds. The presence of NAS intramammary infections was found to be significantly related to the farming systems, composite cow milk somatic cell count (SCC), parity and days in milk (DIM). Significantly more NAS were isolated from primiparous and from older cows. This knowledge could assist with the management of NAS on dairy farms.The National Research Foundation of South Africa and Milk SA.http://www.ojvr.orgam2023Production Animal Studie

Antimicrobial resistance among Escherichia coli isolates from dogs presented with urinary tract infections at a veterinary teaching hospital in South Africa

Background This study investigated the burden and predictors of canine E. coli urinary tract infections (UTI) and antimicrobial resistance among dogs presented at a veterinary teaching hospital in South Africa, 2007–2012. Methods The Cochran-Armitage trend test was used to investigate temporal trends while logistic regression models were used to investigate predictors (age, sex, breed, year) of E. coli infections and antimicrobial resistance (AMR). Results A total of 22.3% (168/755) of the urinary specimens tested positive for E. coli. A significant (p = 0.0004) decreasing temporal trend in the percentage of E. coli positive isolates was observed over the study period. There were high levels of AMR to penicillin-G (99%), clindamycin (100%), tylosine (95%), cephalothin (84%) but relatively low levels of resistance to enrofloxacin (16%), orbifloxacin (21%). Almost all (98%, 164/167) the isolates exhibited multidrug resistance (MDR), while only 11% (19/167) and 2% (4/167) exhibited extensive drug resistance (XDR) and pan-drug resistance (PDR), respectively. Conclusions Although, the risk of E. coli UTI declined during the study period, the risk of AMR increased. The high levels of AMR and MDR as well as the presence of XDR and PDR is concerning as these have the potential of affecting prognosis of UTI treatments

Antimicrobial resistance patterns and biofilm formation of coagulase negative Staphylococcus species isolated from cow milk samples

Increased prevalence of antimicrobial resistance, treatment failure, and financial losses have been reported in dairy cattle with coagulase-negative Staphylococcus (CoNS) clinical mastitis. However, studies on CoNS are limited in South Africa. Therefore, the objectives of this study were to investigate the antimicrobial resistance patterns and biofilm formation in CoNS isolated from cow milk samples submitted to the Onderstepoort Milk Laboratory. A total of 142 confirmed CoNS isolates were used for this study. Isolates were subjected to the tissue culture plate method for biofilm formation testing and antimicrobial susceptibility testing against a panel of 11 antimicrobials using the disk diffusion method. Biofilm formation was identified in 18% of CoNS tested. Staphylococcus chromogenes (11%) had the highest proportion of biofilm formation followed by S. haemolyticus 4.0% and S. epidermidis, S. hominis, S. xylosus, and S. simulans with 1% respectively. Ninety percent (90%) of CoNS isolates were resistant to at least one antimicrobial (AMR) and 51% were multidrug-resistant (MDR). Resistance among CoNS was the highest to ampicillin (90%) and penicillin (89%), with few isolates resistant to cefoxitin and vancomycin, 9% respectively. The most common resistance patterns among the CoNS was penicillin-ampicillin (16%) and penicillin-ampicillin-erythromycin (10%). Forty-two percent (42%) of biofilm positive CoNS were MDR. At the species level, MDR was common among S. epidermis (65%), S. chromogenes (52%) and S. haemolyticus (44%). In conclusion, biofilm formation was uncommon among the MDR-CoNS isolates in this study suggesting that biofilm formation is not a major contributing factor to antimicrobial resistance in this study. In addition, most CoNS isolates in this study were _-lactams resistant. This is concerning as penicillins are used commonly by dairy farmers in treatment of mastitis in South Africa. Nonetheless, the role of antimicrobial use practice in the development of resistance in subclinical mastitis in the dairy industry should be investigated.Dissertation (MSc)--University of Pretoria, 2019.Paraclinical SciencesMScUnrestricte

The molecular epidemiology and antimicrobial resistance of Staphylococcus pseudintermedius canine clinical isolates submitted to a veterinary diagnostic laboratory in South Africa

Staphylococcus pseudintermedius is an important cause of clinical infections in small-animal-veterinary medicine. Evolutionary changes of strains using multilocus sequence typing (MLST) have been observed among S. pseudintermedius in European countries and the United States. However, there are limited or no studies on the detection of methicillin resistant Staphylococcus pseudintermedius (MRSP) and predominating MLST strains in South Africa. Therefore, this study aimed to determine the molecular epidemiology of S. pseudintermedius in South Africa. Twenty-six, non-duplicate, clinical isolates from dogs were obtained as convenience samples from four provinces in South Africa. The Kirby Bauer disk diffusion test was used to determine antimicrobial susceptibility. We used Resfinder and the Comprehensive Antibiotic Resistance Database (CARD) to detect antimicrobial resistance genes. Virulence genes were identified using the virulence factor database and Basic Local Alignment Search Tool (BLASTN) on Geneious prime. geoBURST analysis was used to study relationships between MLST. Finally, the maximum likelihood phylogeny was determined using Randomized Axelerated Maximum Likelihood (RAxML). Twenty-three isolates were confirmed as S. pseudintermedius of which 14 were MRSP. In addition to β-lactam antimicrobials, MRSP isolates were resistant to tetracycline (85.7%), doxycycline (92.8%), kanamycin (92.8%), and gentamicin (85.7%). The isolates harbored antimicrobial resistance genes (tetM, ermB, drfG, cat, aac(6’)-Ie-aph(2”)-Ia, ant(6)-Ia, and aph(3’)-III) and virulence genes (AdsA, geh, icaA, and lip). MLST analysis showed that ST2228, ST2229, ST2230, ST2231, ST2232, ST2318, ST2326 and ST2327 are unique sequence types in South Africa. Whereas, previously reported major STs including ST45, ST71, ST181, ST551 and ST496 were also detected. The geoBURST and phylogenetic analysis suggests that the isolates in South Africa are likely genetically related to isolates identified in other countries. Highly resistant MRSP strains (ST496, ST71, and ST45) were reported that could present challenges in the treatment of canine infections in South Africa. Hence, we have gained a better understanding of the epidemiology of MRSP in the African continent, the genes involved in resistance and virulence factors associated with these organisms