scholarly journals In vitro antimicrobial susceptibility of Staphylococcus aureus strains from dairy herds in KwaZulu-Natal

Author(s):  
T. Schmidt

Staphylococcus aureus is 1 of the most important causes of bovine mastitis and is responsible for significant economic losses to the dairy industry worldwide. One of the principal approaches used in treating intramammary infections is the administration of antimicrobials. Due to the propensity of S. aureus to develop resistance, antimicrobial susceptibility monitoring is necessary to ensure that treatment regimens are effective. As part of this investigation, 90 S. aureus strains isolated from mastitis cases submitted to Allerton Provincial Veterinary Laboratory during 2008 and 2009 were evaluated for their susceptibility to a panel of 10 antimicrobials. Only 8 of the 90 S. aureus isolates tested (8.9 %) were found to be susceptible to all of the antimicrobials evaluated. A very high level of resistance to the beta-lactam antibiotics was noted: 47.8 % of the isolates were resistant to penicillin and 65.6 % were resistant to ampicillin. Minimal resistance to oxacillin, cephalothin and trimethoprim-sulfamethoxazole (1.1 %) was found. Seventeen (18.9 %) of the isolates tested were found to be resistant to 3 or more antimicrobials. The need for vigilant monitoring of bacterial resistance trends in the dairy industry is warranted as the potential public health implications are significant.

Author(s):  
Cosmina Bouari ◽  
George Nadas ◽  
Flore Chirila ◽  
Sorin Rapuntean ◽  
Cornel Catoi ◽  
...  

 Mastitis in cows, one of the most common and economically important infectious diseases of dairy cattle, all over the world, with significant impact due to economic losses, occurs when the udder becomes inflamed because the leukocytes are released into the mammary gland usually in response to bacteria invasion of the teat canal.The main objective of this study was to evaluate the in vitro antimicrobial susceptibility of bacteria isolated from milk in order to design specific control programs for bovine mastitis in this area.A total of 204 milk samples aseptically collected both from farms and private owners were processed during May 2014 and March 2016 within the Microbiology Laboratory of the Faculty of Veterinary Cluj-Napoca, Romania. The microbiological examination was carried out by inoculation on blood agar and MacConkey medium. After the overnight incubation in aerobic conditions, the identification of the isolates was performed using microscopic, cultural and biochemical methods. Biochemical identification was based on API 20 Biomerieux system. Susceptibility to antibiotics was evaluated using Kirby Bauer disk diffusion method on Mueller-Hinton agar; the antibiotics were represented by Amoxicillin and Clavulanic Acid, Ceftiofur, Florfenicol, Mastidiscs, Enrofloxacin, Penicillin and Tetracycline.Staphylococcus spp. was the most common isolated pathogen, in 54.9% of the specimens, followed by Streptococcus spp. in 20.1%, Escherichia coli in 10.78%, Klebsiella spp. in 8.34%, Bacillus spp. in 5.88%. The most frequent associations were represented by staphylococci-streptococci in 62.7% of the samples, followed by streptococci-bacillus in 19.8% of the samples. The most important etiological agents identified were Staphylococcus aureus, S uberis, Streptococcus agalactiae, and Escherichia coli. Antimicrobial susceptibility test for the total isolates revealed good sensitivity to Enrofloxacin, Mastidiscs and Amoxicillin and Clavulanic Acid. Resistance was observed for Penicillin and Tetracycline.The major mastitis pathogens identified was Staphylococcus aureus, while recurrent mastitis treatment was based on systemic and local administration of Enrofloxacin and Mastidiscs respectively.


2014 ◽  
Vol 34 (10) ◽  
pp. 947-952 ◽  
Author(s):  
Daniele C. Beuron ◽  
Cristina S. Cortinhas ◽  
Bruno G. Botaro ◽  
Susana N. Macedo ◽  
Juliano L. Gonçalves ◽  
...  

The objective of this study was to evaluate herd management practices and mastitis treatment procedures as risk factors associated with Staphylococcus aureus antimicrobial resistance. For this study, 13 herds were selected to participate in the study to evaluate the association between their management practices and mastitis treatment procedures and in vitro antimicrobial susceptibility. A total of 1069 composite milk samples were collected aseptically from the selected cows in four different periods over two years. The samples were used for microbiological culturing of S. aureus isolates and evaluation of their antimicrobial susceptibility. A total of 756 samples (70.7%) were culture-positive, and S. aureus comprised 27.77% (n=210) of the isolates. The S. aureus isolates were tested using the disk-diffusion susceptibility assay with the following antimicrobials: ampicillin 10mg; clindamycin 2μg; penicillin 1mg; ceftiofur 30μg; gentamicin 10mg; sulfa-trimethoprim 25μg; enrofloxacin 5μg; sulfonamide 300μg; tetracycline 30μg; oxacillin 1mg; cephalothin 30μg and erythromycin 5μg. The variables that were significantly associated with S. aureus resistance were as follows: the treatment of clinical mastitis for ampicillin (OR=2.18), dry cow treatment for enrofloxacin (OR=2.11) and not sending milk samples for microbiological culture and susceptibility tests, for ampicillin (OR=2.57) and penicillin (OR=4.69). In conclusion, the identification of risk factors for S. aureus resistance against various mastitis antimicrobials is an important information that may help in practical recommendations for prudent use of antimicrobial in milk production.


Antibiotics ◽  
2021 ◽  
Vol 10 (6) ◽  
pp. 637
Author(s):  
Vladimir Gostev ◽  
Olga Kalinogorskaya ◽  
Ksenia Ivanova ◽  
Ekaterina Kalisnikova ◽  
Irina Lazareva ◽  
...  

Selective pressure of beta-lactams is thought to be responsible for mutation selection in methicillin-susceptible Staphylococcus aureus (MSSA). We used next-generation sequencing to compare the genomes of beta-lactamase-positive (SA0707) and -negative (SA0937) MSSA isolates with their derivatives obtained after selection with oxacillin, ceftaroline, or meropenem. Selection with oxacillin and ceftaroline caused a rapid and significant (6–8 times) increase in the minimum inhibitory concentration (MICs) of oxacillin, penicillin, amoxicillin/clavulanate, and ceftaroline against the derivatives of both isolates, associated with growth impairment. Selection with meropenem caused a limited increase in the MICs of all beta-lactams against both isolates. During the initial stages of selection (after 5–15 passages), mutations were detected only in some reads, which indicated the heterogeneity of the population; however, during the later stages, either the population reversed to the wild type or fixation of the mutation was observed in the entire population. Selection with different beta-lactams caused diverse mutational events, but common mutations were detected in gdpP, all penicillin-binding proteins, cell wall regulators (vraST, graR), and deletions in the promoter region of pbp4. Therefore, the disk diffusion test with cefoxitin does not reveal resistance associated with these mechanisms in some cases, which can lead to the failure of beta-lactam therapy.


2021 ◽  
Vol 23 (1) ◽  
pp. 334
Author(s):  
Lei Zhang ◽  
Xin Ye ◽  
Yan Jia ◽  
Manling Cheng ◽  
Dangjin Wu ◽  
...  

Staphylococcus aureus is a vital bovine mastitis pathogen causing huge economic losses to the dairy industry worldwide. In our previous studies, leukotoxin ED (LukED) was detected in most S. aureus strains isolated from bovine mastitis. Here, four single-chain fragment variables (scFvs) (ZL8 and ZL42 targeting LukE, ZL22 and ZL23 targeting LukD) were obtained using purified LukE and LukD proteins as the antigens after five rounds of bio-panning. The complementarity-determining region 3 (CDR3) of the VH domain of these scFvs exhibited significant diversities. In vitro, the scFvs significantly decreased LukED-induced cell killing by inhibiting the binding of LukED to chemokine receptors (CCR5 and CXCR2) and reduced the death rates of bovine neutrophils and MAC-T cells caused by LukED and S. aureus (p < 0.05). In an S. aureus-induced mouse mastitis model, histopathology and MPO results revealed that scFvs ameliorated the histopathological damages and reduced the infiltration of inflammatory cells (p < 0.05). The ELISA and qPCR assays showed that scFvs reduced the transcription and expression levels of Tumor Necrosis Factor-alpha (TNF-α), interleukin-1β (IL-1β), IL-6, IL-8 and IL-18 (p < 0.05). The overall results demonstrated the protective anti-inflammatory effect of scFvs in vitro and in vivo, enlightening the potential role of scFvs in the prevention and treatment of S. aureus-induced mastitis.


Antibiotics ◽  
2020 ◽  
Vol 9 (8) ◽  
pp. 469
Author(s):  
Leta Elias ◽  
Ajay S. Balasubramanyam ◽  
Olena Y. Ayshpur ◽  
Iryna U. Mushtuk ◽  
Nataliya O. Sheremet ◽  
...  

Bovine mastitis is the predominant cause for antimicrobial use on dairy farms and is a major source of economic losses in the dairy industry. In this study, the antimicrobial susceptibility profiles of common mastitis-causing pathogens, Staphylococcus aureus (n = 62), Streptococcus agalactiae (n = 46), and Escherichia coli (n = 129), were determined for dairy cattle with mastitis across 142 Ukrainian farms. The results showed that there were more gentamicin resistant S. aureus isolates (16.95%) identified in this study than previously reported for Ukrainian dairy cattle. Moreover, low levels of amoxicillin susceptibly (13.51%) were observed for St. agalactiae, which contrasted a previous study showing susceptibility levels of >50%. St. agalactiae resistance to tetracycline was observed in 80% of the isolates. Cephalosporin use was most ineffective against E. coli, with 43.27–56% of the isolates exhibiting this resistant trait. Overall, this study performed a preliminary analysis of antimicrobial resistance on mastitis isolates from Ukrainian farms. However, given the limited numbers of the isolates tested in this study and that the publications on antimicrobial resistance in animal husbandry of Ukraine are very few, more extensive investigations are needed to comprehensively examine susceptibility patterns of mastitis-causing pathogens in dairy cattle in Ukraine.


2001 ◽  
Vol 45 (12) ◽  
pp. 3422-3426 ◽  
Author(s):  
Siddhartha Roychoudhury ◽  
Tracy L. Twinem ◽  
Kelly M. Makin ◽  
Mark A. Nienaber ◽  
Chuiying Li ◽  
...  

ABSTRACT The in vitro development of resistance to the new nonfluorinated quinolones (NFQs; PGE 9262932, PGE 4175997, and PGE 9509924) was investigated in Staphylococcus aureus. At concentrations two times the MIC, step 1 mutants were isolated more frequently with ciprofloxacin and trovafloxacin (9.1 × 10−8 and 5.7 × 10−9, respectively) than with the NFQs, gatifloxacin, or clinafloxacin (<5.7 × 10−10). Step 2 and step 3 mutants were selected via exposure of a step 1 mutant (selected with trovafloxacin) to four times the MICs of trovafloxacin and PGE 9262932. The step 1 mutant contained the known Ser80-Phe mutation in GrlA, and the step 2 and step 3 mutants contained the known Ser80-Phe and Ser84-Leu mutations in GrlA and GyrA, respectively. Compared to ciprofloxacin, the NFQs were 8-fold more potent against the parent and 16- to 128-fold more potent against the step 3 mutants. Mutants with high-level NFQ resistance (MIC, 32 μg/ml) were isolated by the spiral plater-based serial passage technique. DNA sequence analysis of three such mutants revealed the following mutations: (i) Ser84-Leu in GyrA and Glu84-Lys and His103-Tyr in GrlA; (ii) Ser-84Leu in GyrA, Ser52-Arg in GrlA, and Glu472-Val in GrlB; and (iii) Ser84-Leu in GyrA, Glu477-Val in GyrB, and Glu84-Lys and His103-Tyr in GrlA. Addition of the efflux pump inhibitor reserpine (10 μg/ml) resulted in 4- to 16-fold increases in the potencies of the NFQs against these mutants, whereas it resulted in 2-fold increases in the potencies of the NFQs against the parent.


2020 ◽  
Vol 98 (Supplement_4) ◽  
pp. 259-259
Author(s):  
Yuhui Zheng ◽  
Shengli Li

Abstract Bovine mastitis is one of the major diseases which directly affects the milk production performance and it causes huge economic losses in the dairy industry. Bacterial infection is the main risk factor of bovine mastitis and the antibiotic therapy is the primary choice to control the disease. However, persistence use of antibiotic increases the incidence of bacterial resistance and traces of antibiotic residues in animal products. Lactobacillus casei Zhang is one of the probiotics with multiple biological functions, which has certain bacteriostatic effect on pathogenic microorganism. The purpose of this study was to explore the effect of Lactobacillus casei Zhang (L. casei Zhang) on the prevention of E. coli-induced milk-blood barrier damage. Bovine mammary epithelial cells (BMECs) were used to establish a milk-blood model and Control group (PBS), E. coli group, and L. casei Zhang pretreatment plus E. coli group were set up respectively. The results showed that: L. casei Zhang could significantly reduce the increase of LDH release caused by E. coli treatment (P&lt; 0.05). And it can also significantly reduce the decrease of transmembrane resistance of monolayer cells caused by E. coli treatment (P&lt; 0.05). In addition, L. casei Zhang could significantly reduce the expression of tight junction proteins ZO-1, Claudin-1, Claudin-4 and Occludin (P &lt; 0.05). In conclusion, L. casei Zhang could effectively improve the damage of the blood-milk barrier caused by E. coli and could protect BMECs during bacterial infection.


2017 ◽  
Vol 11 (4) ◽  
pp. 219-225
Author(s):  
Mylena Karoline Valmorbida ◽  
Danielle Correia dos Santos Carneiro ◽  
Keila Catarina Prior ◽  
Eliete Griebeler ◽  
Marcella Zampoli Troncarelli ◽  
...  

2021 ◽  
Vol 22 (16) ◽  
pp. 8840
Author(s):  
Katharina Mayer ◽  
Martin Kucklick ◽  
Helene Marbach ◽  
Monika Ehling-Schulz ◽  
Susanne Engelmann ◽  
...  

Within-host adaptation is a typical feature of chronic, persistent Staphylococcus aureus infections. Research projects addressing adaptive changes due to bacterial in-host evolution increase our understanding of the pathogen’s strategies to survive and persist for a long time in various hosts such as human and bovine. In this study, we investigated the adaptive processes of S. aureus during chronic, persistent bovine mastitis using a previously isolated isogenic strain pair from a dairy cow with chronic, subclinical mastitis, in which the last variant (host-adapted, Sigma factor SigB-deficient) quickly replaced the initial, dominant variant. The strain pair was cultivated under specific in vitro infection-relevant growth-limiting conditions (iron-depleted RPMI under oxygen limitation). We used a combinatory approach of surfaceomics, molecular spectroscopic fingerprinting and in vitro phenotypic assays. Cellular cytotoxicity assays using red blood cells and bovine mammary epithelial cells (MAC-T) revealed changes towards a more cytotoxic phenotype in the host-adapted isolate with an increased alpha-hemolysin (α-toxin) secretion, suggesting an improved capacity to penetrate and disseminate the udder tissue. Our results foster the hypothesis that within-host evolved SigB-deficiency favours extracellular persistence in S. aureus infections. Here, we provide new insights into one possible adaptive strategy employed by S. aureus during chronic, bovine mastitis, and we emphasise the need to analyse genotype–phenotype associations under different infection-relevant growth conditions.


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