scholarly journals Investigation of Methicillin Resistance and Panton-Valentine Leukocidin in Staphylococci Isolated from Bovine Mastitis

2018 ◽  
Vol 44 (1) ◽  
pp. 9
Author(s):  
Cansu Gezgen ◽  
Esra Seker

Background: Mastitis, which is inflammation of the mammary gland, is among the most important diseases in dairy herds resulting in reductions of milk yield and milk quality. Although several groups of microorganisms have been reported as etiological agents of mastitis, Staphylococci are the most frequently isolated bacteria from bovine mastitic milk samples. The aim of this study was to isolate the Staphylococcus species from bovine mastitis, investigate the mecA ve pvl genes in isolated species by PCR and determine the antibiotic resistance of methicillin resistant strains to some antibiotics commonly used in veterinary field.Materials, Methods & Results: In the present study, 972 half-udder milk samples (n = 757 CMT positive, n = 215 CMT negative) were used from 251 lactating cows from 34 different enterprises located center town and villages of Ödemiş, İzmir. Ten microliters of each milk sample was inoculated onto Columbia blood agar, containing 7% of sheep blood and incubated under aerobic conditions for 24-48 h at 37°C. The certain identification of Staphylococcus isolates was achieved using Crystal™ Identification Systems Gram-Positive ID kit. Bacterial DNAs were extracted from all strains using boiling method and strains were screened for the presence of 16S rDNA, mecA and pvl genes by PCR. The antimicrobial resistance of MRS species was determined by using disc diffusion method. A total of 182 (18.72%) Staphylococcus strains were isolated from 972 half-udder milk samples. Of 182 Staphylococcus strains, 137 (75.27%) and 45 (24.73%) were detected as CPS and CNS, respectively. Among the 11 different Staphylococcus species, S. intermedius (42.30%) was the most common species isolated, followed by S. aureus (32.97%) and S. saprophyticus (10.99%). The mecA positivity was found in only 4 (2.2%) S. intermedius strains, while pvl toxin gene was determined in none of the strains. Four MR S. intermedius strains were resistant to oxacillin and cefoxitin. The resistance was also found to erythromycin (50%), rifampicin (25%), gentamicin (25%), tetracycline (25%) and trimethoprim/sulfamethoxazole (25%) in the isolates.Discussion: In this study, S. intermedius had the highest isolation rate and this finding was considered remarkable. Generally, in mastitis diagnostics all CPS isolates are classified as S. aureus. In our study, the certain identification of all CPS may explain the high isolation rate of S. intermedius. The sampling method may also be the reason of higher isolation rate for S. intermedius in accordance with the most common ones causing mastitis. All of mecA positive strains were S. intermedius and this was the another remarkable finding of our study. Because similar result was seen in only one study from Korea, while the investigation finding related to MR S. intermedius was not determined in Turkey. However, the mecA positivity found in our study was lower than the other author’s isolation rate. The difference between the sample size, geographical variations and diversity in strains may be the causes of this discrepancy. It was investigated pvl toxin gene in 182 Staphylococcus strains by PCR and found this gene in none of strains. According to this finding, it was considered that pvl gene may have not an efficient role in the pathogenesis of mastitis in terms of sampled animals and sampling area. Antibiotic resistance of 4 MR S. intermedius strains against various antibiotics commonly used in Turkey was investigated. Of methicillin resistant strains, 2, 1 and 1 were resistant to 3, 6 and 5 antibiotics, respectively. It was considered that geographical differences, number of tested isolates and diversity in MR strains may be effective on the antibiotic resistance levels. This is the first study showing the presence of mecA gene in S. intermedius strains isolated from bovine mastitic milk samples in Turkey.

2018 ◽  
Vol 20 (87) ◽  
pp. 112-115
Author(s):  
T. Kozytska ◽  
T. Garkavenko

The frequency of Staphylococcus aureus resistance to antibiotics (AB) has significant differences in different countries and geographical regions. The main problem is represented by strains of methicillin-resistant staphylococcus (MRSA), resistant to many ABs. Often, MRSA is transmitted to humans through animal food products. Purpose: investigate animal foods for S. aureus and detect methicillin-resistant strains. Food research was carried out according to ISO 6888-1, ISO 6888-2, MW 15.2-5.3-004:2007, State standard 10444.2. The material for further research was the isolates of S. aureus isolated from different groups of food products of animal origin. Disks containing methicillin (30 mg) and oxacillin (5 mg) and Müller-Hinton nutrient medium were used to determine the sensitivity to AB. The research was conducted by disc diffusion method in accordance with the Methodological Guidelines «Determination of microorganism sensitivity to antibacterial preparations» dated 12/25/2014. In addition, the data of the state veterinary report on the study of milk of cows and determination of antibiotic resistance of cultures isolated from it in 2017 were analyzed. A study on the presence of coagulase-positive Staphylococcus was carried out in food products of animal origin during 2016–2017. During this period, 58 isolates of S. aureus were identified, of which 7 were classified as MRSA, which was an average of 12.1 % of all isolated cultures. The highest amount of MRSA is isolated from chicken meat – 29.0% and meat products – 20.0%, ready-to-eat fish – 16.7% and meat mincer – 6.1%. The 2017 report on antibiotic resistance from all regions of Ukraine was also analyzed for the detection of S. aureus strains resistant to oxacillin. These data were available only for S. aureus isolated from milk from cows suffering from subclinical mastitis. Of the 47 strains of S. aureus, 22 (46.8%) were methicillin-resistant. MRSA poses a serious threat to the health of humans and animals and may be transmitted through food. There is no monitoring system in Ukraine on the detection of resistant strains in food products. A single list of ABs according to which laboratories conducted sensitivity tests was not approved. Therefore, in most cases, the sensitivity of Staphylococcus to oxacillin/methicillin is not determined. We found that, on average, MRSA from animal foods of animal origin in Ukraine is found in 12.1% of cases. Milk of animals, that suffer from subclinical forms of mastitis, can serve as a source of MRSA for humans, because it was detected in 46.8% of cases.


2021 ◽  
Vol 8 ◽  
Author(s):  
Tariq Ali ◽  
Kamran ◽  
Abdur Raziq ◽  
Inamullah Wazir ◽  
Rafi Ullah ◽  
...  

Mastitis is the most prevalent disease of dairy animals, imparting huge economic losses to the dairy industry. There is always a dire need to monitor the prevalence of mastitis, its bacteriology, and evaluation of antimicrobial susceptibilities for mastitis control and prevention. Therefore, the objectives of this study were to investigate: (i) the prevalence of mastitis in cattle and buffaloes; (ii) identification of bacteria associated with mastitis; (iii) antimicrobial susceptibility of bacterial isolates. Milk samples (n = 1,566) from cattle (n = 1,096) and buffaloes (n = 470) were processed for detection of mastitis using the California mastitis test in the year 2018–19. A total of 633 mastitic milk samples were further processed for bacteriology and antimicrobial susceptibility testing by the disc diffusion method. Overall, the prevalence of clinical and subclinical mastitis was 17 and 57% in both species. Clinical mastitis was higher in cattle (20%) compared to buffaloes (11%), whereas subclinical was higher in buffaloes (66%) than cattle (53%). Besides, month-wise prevalence was higher in hot and humid months in both species. Staphylococci spp. (34%) were the most predominant bacterial isolates from mastitic milk, followed by Escherichia coli (19.4%), Streptococci spp. (9%), and Klebsiella spp. (8%). Most of the bacteria were susceptible to gentamicin (92%) and enrofloxacin (88%), when a panel of 16 different antimicrobials was tested. Nevertheless, most of the isolates were resistant to sulphamethoxazole (99%), lincomycin (98%), oxytetracycline (89%), ampicillin (86%), and doxycycline (85%). This study concludes a high prevalence of mastitis caused by Staphylococcal spp. in cattle and buffaloes belonging to the northwest of Pakistan, and gentamicin and enrofloxacin might be appropriate antimicrobial agents in the treatment of bovine mastitis.


2021 ◽  
Vol 14 (6) ◽  
pp. 592
Author(s):  
Pavarish Jantorn ◽  
Hawaree Heemmamad ◽  
Tanawan Soimala ◽  
Saowakon Indoung ◽  
Jongkon Saising ◽  
...  

Staphylococcus pseudintermedius is a zoonotic pathogen that can cause life-threatening infections in animals and humans. The study of methicillin-resistant S. pseudintermedius (MRSP) and its ability to produce biofilms is important to select the most suitable treatment. The prevalence and characteristics of S. pseudintermedius isolated from dogs admitted at the Veterinary Teaching Hospital, Prince of Songkla University, Thailand were assessed. Results showed that 28.30% (15/53) of the isolates were MRSP. Amplification of the mecA gene was observed in 93.33% (14/15) MRSP. Methicillin-resistant strains revealed co-resistant patterns against other antibiotics, including chloramphenicol, clindamycin, tetracycline, clarithromycin, ciprofloxacin, and trimethoprim. In this study, all bacterial isolates produced biofilms, while 90.55% of S. pseudintermedius isolates were strong or moderate biofilm producers. Most (45–60%) of the resistant strains were strong biofilm producers, while the correlation between biofilm production and antibiotic resistance was not statistically significant. This is the first study in southern Thailand to investigate the drug-resistant profile of S. pseudintermedius and its ability to form biofilm. The results will contribute to a better understanding of the emergence and prevalence of antimicrobial resistance in S. pseudintermedius.


Author(s):  
Jayaprabha Chockalingam ◽  
Akshaya Balasubramanian ◽  
Jai Santhosh K. Krishnasamy ◽  
Karthika Balasundaram ◽  
Kavin Kumar Sivasamy ◽  
...  

Author(s):  
D.J. Vatalia ◽  
B.B. Bhanderi ◽  
V.R. Nimavat ◽  
M.K. Jhala

Background: Mastitis, the inflammation of parenchyma of mammary gland is frequently considered to be costliest and complex disease prevalent in India. Mastitis is caused by pathogens like Staphylococcus spp., Streptococcus spp., Mycoplasma bovis, E. coli, Klebsiella spp., Citrobacter spp., Enterobacter spp. and Entercoccus. The treatment of mastitis in animals is carried out using antibiotics. Treatment failure in mastitis is due to increased antibiotic resistance of mastitis pathogens and also due to indiscriminate use of antibiotics without testing in vitro antibiotic sensitivity test against causal organisms. In comparison to cultural method, PCR assays takes less time for detection of bacteria from the mastitis milk samples. Present research work was carried out regarding isolation, identification and multiple drug resistance profile of clinical bovine mastitis associated pathogens using conventional as well as molecular approach. Methods: In the present study, 73 mastitis milk samples were collected from Anand and Panchmahal district of Gujarat. The milk samples were subjected for cultural isolation and DNA extraction for identification of bacteria by cultural and PCR method. Antimicrobial sensitivity pattern of the isolates were carried by disc diffusion method and isolates were categorized in multiple drug resistant. Result: In the present study, Out of 73 mastitis milk samples collected from cows 48 (65.75%) cows were positive for bacterial isolation and S. aureus was the most predominant bacterial species. PCR from the mastitis milk additionally detected bacteria in culturally negative milk samples. Most sensitive drug was gentamicin and most of the isolates (90.19%) showed the multiple drug resistance for the two to nine drugs with 0.1 to 0.6 multiple antibiotic resistance index.


Author(s):  
Abolfazl Jafari-Sales ◽  
Zahra Sadeghi Deylamdeh ◽  
Afsoon Shariat

Introduction: Staphylococcus aureus causes a wide range of infections and as a multivalent pathogen is one of the causative agents of nosocomial and community infections. Therefore, the aim of this study was to identify and determine the pattern of antibiotic resistance of methicillin-resistant Staphylococcus aureus (MRSA) isolates from patients in hospitals and medical centers in Marand city and also to evaluate the presence of mecA gene. Materials and Methods: In this cross-sectional descriptive study, 385 samples of S. aureus were collected from different clinical samples of patients in hospitals and medical centers of Marand city. S. aureus was identified using standard biochemical methods.  Methicillin resistance was determined by disk diffusion method in the presence of oxacillin and cefoxitin. The pattern of antibiotic resistance of the strains was determined by disk diffusion method and according to CLSI recommendation and also PCR method was used to evaluate the frequency of MecA gene. Results: In the present study, out of 385 samples of S. aureus, 215 (55.84%) samples were methicillin resistant. PCR results for mecA gene showed that 110 samples had mecA gene.  The highest antibiotic resistance was observed against penicillin (100%) and erythromycin (83.63%). Most MRSA were isolated from urine and wound samples. Conclusion: The results of this study indicate the prevalence of methicillin-resistant species and also the increase in antibiotic resistance of MRSA to various antibiotics.  Therefore, in order to prevent increased resistance to other antibiotics, it is recommended to avoid inappropriate use of antibiotics.


Author(s):  
Somaye Delfani ◽  
Faranak Rezaei ◽  
Setareh Soroush ◽  
Pegah Shakib

Background: Methicillin-resistant coagulase-negative staphylococci is responsible for hospital and community-acquired infections. Objective: This study aimed to investigate the antibiotic-resistance patterns, antibiotic-resistance genes, namely, ermA, ermB, ermC, blaZ, msrA, tetK, tetM, mup, and vanA, biofilm formation, and prevalence of different SCCmec types among the Staphylococcus cohniistrains isolated from clinical samples in Tehran, Iran. Methods: In this study,S. cohniiisolates were screened from the clinical samples from March 2012 to February 2013 in Tehran, Iran.Antimicrobial susceptibility test and inducible clindamycin resistance were evaluated by disc diffusion method, andresistance genes were examined using Polymerase Chain Reaction (PCR) assays. Then, biofilm formation assay was analyzed by Microtiter-plate test to detect the icaA and icaDgenes. The SCCmec and the Arginine Catabolite Mobile Element (ACME) typing were performed using the PCRmethod. Results: FromtwentyS. cohnii, all isolates were resistant to cefoxitin. 95% of the S. cohnii was defined as multidrug resistance (MDR)strains. The ermB, ermC, and vanA genes were not detected in any isolates; however, the blaZ gene had the highest frequency.95% of the S. cohnii isolates produced biofilm. Also, 4 SCCmec types, including V, IV, III+ (C2), VIII+ (AB1), were identified. Therefore, the majority of SCCmec were untypable. Based on the ACME typing, arcA and opp3 genes were positive in 13 (65%) and 1 (5%) isolates, respectively. Conclusion: Due to the high antimicrobial resistance and the spread of untypableSCCmecamong the isolates studied, the control and treatment of methicillin-resistantS. cohnii in hospitals and public health centers is a significant concern.


2007 ◽  
Vol 70 (12) ◽  
pp. 2764-2768 ◽  
Author(s):  
DANIEL LOETO ◽  
M. I. MATSHEKA ◽  
B. A. GASHE

The prevalence, antibiotic resistance, and enterotoxigenic potential of Staphylococcus aureus strains from different anatomical sites on food handlers in Gaborone, Botswana, were determined. Of a total of 200 food handlers tested, 115 (57.5%) were positive for S. aureus. Of the 204 S. aureus isolates, 63 (30.9%), 91 (44.6%), and 50 (24.5%) were isolated from the hand, nasal cavity, and face, respectively, and 43 (21%) of the isolates were enterotoxigenic. The most prevalent enterotoxin was type A, which accounted for 34.9% of all the enterotoxigenic strains, and enterotoxin D was produced by the fewest number of strains (9.3%). Resistance to methicillin was encountered in 33 (22.4%) of the penicillin G–resistant isolates, and 9 (27.3%) of these methicillin-resistant isolates also were resistant to vancomycin. Nineteen antibiotic resistance profiles were determined, and the nasal cavity had the highest diversity of resistance profiles. The nasal cavity also had the highest number of resistant strains, 77 (53%), whereas the hand and face had 49 (32%) and 24 (16.0%) resistant strains, respectively. To reduce the Staphylococcus carriage rate among food handlers, training coupled with a commitment to high standards of personal and environmental hygiene is recommended.


2020 ◽  
Vol 2020 ◽  
pp. 1-9
Author(s):  
Justine Fri ◽  
Henry A. Njom ◽  
Collins N. Ateba ◽  
Roland N. Ndip

Thirty-three (33) isolates of methicillin-resistant Staphylococcus aureus (MRSA) from healthy edible marine fish harvested from two aquaculture settings and the Kariega estuary, South Africa, were characterised in this study. The phenotypic antimicrobial susceptibility profiles to 13 antibiotics were determined, and their antibiotic resistance determinants were assessed. A multiplex PCR was used to determine the epidemiological groups based on the type of SCCmec carriage followed by the detection of staphylococcal enterotoxin-encoding genes sea-sed and the Panton Valentine leucocidin gene (pvl). A high antibiotic resistance percentage (67–81%) was observed for Erythromycin, Ampicillin, Rifampicin, and Clindamycin, while maximum susceptibility to Chloramphenicol (100%), Imipenem (100%), and Ciprofloxacin (94%) was recorded. Nineteen (58%) of the MRSA strains had Vancomycin MICs of ≤2 μg/mL, 4 (12%) with MICs ranging from 4–8 μg/mL, and 10 (30%) with values ≥16 μg/mL. Overall, 27 (82%) isolates were multidrug-resistant (MDR) with Erythromycin-Ampicillin-Rifampicin-Clindamycin (E-AMP-RIP-CD) found to be the dominant antibiotic-resistance phenotype observed in 4 isolates. Resistance genes such as tetM, tetA, ermB, blaZ, and femA were detected in two or more resistant strains. A total of 19 (58%) MRSA strains possessed SCCmec types I, II, or III elements, characteristic of healthcare-associated MRSA (HA-MRSA), while 10 (30%) isolates displayed SCCmec type IVc, characteristic of community-associated MRSA (CA-MRSA). Six (18%) of the multidrug-resistant strains of MRSA were enterotoxigenic, harbouring the see, sea, or sec genes. A prevalence of 18% (6/33) was also recorded for the luk-PVL gene. The findings of this study showed that marine fish contained MDR-MRSA strains that harbour SCCmec types, characteristic of either HA-MRSA or CA-MRSA, but with a low prevalence of enterotoxin and pvl genes. Thus, there is a need for continuous monitoring and implementation of better control strategies within the food chain to minimise contamination of fish with MDR-MRSA and the ultimate spread of the bug.


2018 ◽  
Vol 85 (1) ◽  
pp. 57-63 ◽  
Author(s):  
Mariela E Srednik ◽  
Valentine Usongo ◽  
Sarah Lépine ◽  
Xavier Janvier ◽  
Marie Archambault ◽  
...  

The study reported in this Research Communication was conducted to characteriseStaphylococcus aureusisolates recovered from mastitic bovine milk from dairy herds in Argentina. A total of 829 mastitic milk samples, both clinical and subclinical, were collected from 21 farms by veterinarians and submitted to the laboratory for testing from which 229S. aureusisolates were recovered, an isolation rate of 28·1%. These isolates were tested for susceptibility to the antibiotics penicillin, erythromycin and clindamycin. Of the 229 isolates, 53 (23·1%) were resistant to penicillin, 31 (13·5%) to erythromycin and 28 (12·2%) to clindamycin. All isolates were negative for themecA,mecC andpvlgenes by PCR. Southernblot hybridisation revealed that theermC gene was located on plasmid bands. Eighty isolates were randomly selected from the 229 for further characterisation. Restriction analysis of chromosomal DNA with Cf9I followed by PFGE of the 80 isolates revealed 23 distinct pulsotypes at 80% similarity. Seven major types (A, B, N, P, S, T, U and V) accounted for 68·7% of these isolates and 12 pulsotypes (A, B, F, G, J, K, M, N, P, S, T and U) occurred on more than one farm indicating genetic diversity within the farms. MLST of a representative isolate from dominant types identified the STs 97 705, 746, 2102 and 2187 with ST97 being the most predominant. Antibiotic susceptibility testing showed that 53·7% of the 80 randomly selected isolates were resistant to at least one of the three antibiotics tested. To our knowledge, this study represents the first large scale molecular studies onS. aureusisolates from dairy farms in Argentina.


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