Characterisation ofStaphylococcus aureusstrains isolated from mastitis bovine milk in Argentina

2018 ◽  
Vol 85 (1) ◽  
pp. 57-63 ◽  
Author(s):  
Mariela E Srednik ◽  
Valentine Usongo ◽  
Sarah Lépine ◽  
Xavier Janvier ◽  
Marie Archambault ◽  
...  
Keyword(s):  
Large Scale ◽  
Susceptibility Testing ◽  
Restriction Analysis ◽  
Bovine Milk ◽  
Dairy Herds ◽  
Antibiotic Susceptibility Testing ◽  
Chromosomal Dna ◽  
Isolation Rate ◽  
Milk Samples ◽  
Mastitic Milk

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.

scholarly journals Distribution of Antibiotic Resistance Genes in Enterococcus spp. Isolated from Mastitis Bovine Milk

2016 ◽  
Vol 66 (3) ◽  
pp. 336-346 ◽  
Author(s):  
Goksel Erbas ◽  
Ugur Parin ◽  
Suheyla Turkyilmaz ◽  
Nese Ucan ◽  
Mehmet Ozturk ◽  
...  
Keyword(s):  
Resistance Genes ◽  
Bovine Milk ◽  
Antibiotic Resistance Genes ◽  
High Rate ◽  
Resistant Isolate ◽  
Chloramphenicol Resistance ◽  
Milk Samples ◽  
Enterococcus Spp ◽  
Pcr Method ◽  
Mastitic Milk

AbstractIn this study, determination of enterococcus species that were isolated from mastitic milk samples, investigation of their susceptibilities to antibiotics and identification of the existence of resistance genes in resistant strains were conducted. The specimens consist of 600 mastitic milk samples that were collected from 242 cows. Isolation of enterococcus was carried out in selective media and 94 (15.6%)Enterococcusspp. were isolated. A total of 94 species of Enterococci were identified using both sequencing and polymerase chain reaction (PCR).Enterococcusspp. isolates belong to 5 different species (E. faecalis, E. faecium, E. durans, E. hirae, E. mundtii) in sequence analysis and 4 different species (E. faecalis, E. faecium, E. durans, E. hirae) were identify by PCR method with specific primers. Analyzing 94 enterococcus strains by antibiotic sensitiveness test a high rate of resistance to tetracycline in 77 (81.9%) isolates was shown. Thetetresistance genes were identified as follows: 54 weretetM positive, 23 weretetK positive and 17 were positive ontetM andtetK. Resistance to erythromycin was established in 27 (28.7%) isolates (25ermB) while the chloramphenicol resistance gene was found in 10 (10.7%) of isolates and thecatgene was identified in nine samples and one isolate was resistant to vancomycin (1.06%) with theVanA gene confirmed. In conclusion, it was shown thatE. faecalishas the biggest role inenterococcusoriginated mastitis and these strains were found to be mostly resistant to tetracycline. One vancomycine resistant isolate that had theVanA gene was also determined.

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
Keyword(s):  
Antibiotic Resistance ◽  
Bovine Mastitis ◽  
Diffusion Method ◽  
Toxin Gene ◽  
Isolation Rate ◽  
Methicillin Resistant ◽  
Milk Samples ◽  
Geographical Variations ◽  
Resistant Strains ◽  
Mastitic Milk

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.

Coagulase-negative staphylococci from non-mastitic bovine mammary gland: characterization ofStaphylococcus chromogenesandStaphylococcus haemolyticusby antibiotic susceptibility testing and pulsed-field gel electrophoresis

2011 ◽  
Vol 79 (2) ◽  
pp. 129-134 ◽  
Author(s):  
Mateja Pate ◽  
Irena Zdovc ◽  
Jana Avberšek ◽  
Matjaž Ocepek ◽  
Andrej Pengov ◽  
...  
Keyword(s):  
Mammary Gland ◽  
Dairy Cows ◽  
Gel Electrophoresis ◽  
Susceptibility Testing ◽  
Clinical Signs ◽  
Pulsed Field Gel Electrophoresis ◽  
Antibiotic Susceptibility Testing ◽  
Bovine Mammary Gland ◽  
Coagulase Negative Staphylococci ◽  
Milk Samples

During routine microbiological examination of milk samples from dairy cows without clinical signs of mastitis, quarter milk samples of 231 dairy cows from 12 herds were investigated for the presence of coagulase-negative staphylococci (CNS). The isolates were identified on the basis of colony morphology, Gram staining, catalase and coagulase test and the commercial kit, API Staph. CNS was detected in 29% (67/231) of the cows. A total of seven CNS species were identified with the most prevalent beingStaphylococcus (Staph.) chromogenes(30%) andStaph. haemolyticus(28·8%), followed byStaph. simulans(11·2%),Staph. xylosus(11·2%),Staph. epidermidis(7·5%),Staph. hyicus(6·3%) andStaph. sciuri(5%). The predominant species,Staph. chromogenesandStaph. haemolyticus, were further characterized by antibiotic susceptibility testing using the agar disc diffusion method (Kirby-Bauer) and by pulsed-field gel electrophoresis (PFGE). Considerable resistance to ampicillin and penicillin was observed in both species. Isolates with identical or highly similar PFGE profiles were detected at the herd level despite a marked heterogeneity seen for both species. On the basis of somatic cell count, absence of clinical signs of inflammation and heterogeneity of genotypes, we assume that CNS isolated in this study could not be considered as important causative agents of the bovine mammary gland inflammation.

Differential titration of plasmin and plasminogen in milk using sandwich ELISA with monoclonal antibodies

1997 ◽  
Vol 64 (1) ◽  
pp. 77-86 ◽  
Author(s):  
DIDIER DUPONT ◽  
CHRISTELLE BAILLY ◽  
JEANNE GROSCLAUDE ◽  
JEAN-CLAUDE COLLIN
Keyword(s):  
Monoclonal Antibodies ◽  
Proteinase Inhibitors ◽  
Sandwich Elisa ◽  
Bovine Milk ◽  
Milk Proteins ◽  
Cross Reactivity ◽  
The Other ◽  
Milk Samples ◽  
Skimmed Milk ◽  
Mastitic Milk

Two sandwich enzyme-linked immunosorbent assays (ELISA) have been developed for quantitation of bovine milk plasminogen and plasmin. The assays used two monoclonal antibodies, one specific for plasminogen and the other specific for plasminogen plus plasmin. Plasmin concentration was obtained by subtracting the first concentration from the second. The assays were sensitive (linear range, 5–75 ng/ml), repeatable (CV, 8 and 5% for plasmin and plasminogen titration respectively), specific (no cross reactivity with the major milk proteins) and directly applicable to skimmed milk with no particular pretreatment of the sample. However, ELISA did not permit complete titration of milk plasminogen and plasmin (only 60–90% could be quantified). This lack of accuracy was due to casein interfering with the plasmin–plasminogen titration by ELISA. Results obtained with ELISA or an enzymic technique on 20 milk samples collected from individual cows milked throughout pregnancy showed that the ELISA was particularly suitable for analysis of late lactation or mastitic milk where proteinase inhibitors interfered with enzymic quantitation.

scholarly journals Characterization of multi-drug resistant Salmonella typhi isolated from Pakistan

2000 ◽  
Vol 124 (1) ◽  
pp. 9-16 ◽  
Author(s):  
P. M. A. SHANAHAN ◽  
K. A. KARAMAT ◽  
C. J. THOMSON ◽  
S. G. B. AMYES
Keyword(s):  
Susceptibility Testing ◽  
Antibiotic Resistance Genes ◽  
Salmonella Typhi ◽  
Pulsed Field Gel Electrophoresis ◽  
Type I ◽  
Drug Resistant ◽  
Antibiotic Susceptibility Testing ◽  
Chromosomal Dna ◽  
Incompatibility Group

Thirty-nine strains of Salmonella typhi, isolated in 1995 from four Districts in Pakistan, Rawalpindi, Islamabad, Kharian and Jehlem, were catalogued and examined. Chromosomal DNA from each isolate was digested with XbaI restriction endonuclease and subjected to pulsed-field gel electrophoresis. Three clonal variants comprising of 17–19 DNA fragments were identified. Antibiotic susceptibility testing identified that 37 of the S. typhi were resistant to chloramphenicol, trimethoprim and ampicillin. These antibiotic resistance genes were found to be located on one of four plasmids belonging to incompatibility group IncHI1 and ranging in size from 150–175 Kb. The genes responsible for this resistance in each case were the chloramphenicol acetyltransferase (CAT) type I, the dihydrofolate reductase (DHFR) type VII and the β-lactamase TEM-1 respectively.

scholarly journals Subclinical intramammary infection does not affect bovine milk ethanol stability

2018 ◽  
Vol 55 (2) ◽  
pp. e135173
Author(s):  
Cristian Marlon de Magalhães Rodrigues Martins ◽  
Juliano Leonel Gonçalves ◽  
Bruna Gomes Alves ◽  
Marcos André Arcari ◽  
Marcos Veiga dos Santos
Keyword(s):  
Bovine Milk ◽  
Subclinical Mastitis ◽  
Milk Composition ◽  
Dairy Herds ◽  
Intramammary Infection ◽  
Milk Samples ◽  
Lactating Cows ◽  
Natural Exposure ◽  
Major Pathogens ◽  
Ethanol Stability

The present study hypothesized that intramammary infection (IMI) might reduce milk ethanol stability (MES), mainly when IMI is caused by major pathogens. Thus, this study evaluated the effect of IMI on bovine MES using a natural exposure experimental design. Ninety-four lactating cows from five dairy herds were selected once they were determined to have an IMI, based on milk bacteriological culturing with positive isolation and somatic cell count (SCC) > 200×103 cells/mL in two out of three composite milk samples collected during three consecutive weeks. After selection, cows were sampled a second time (within two weeks) for evaluation at mammary quarter level (n = 326): milk yield (kg/quarter/day), MES, composition (fat, protein, lactose, casein, total solids and solids-non-fat), and bacteriologic culture. The effect of subclinical mastitis on MES was tested by two models: 1) comparison of healthy vs. infected quarters; and 2) comparison of contralateral mammary quarter within cow. The only milk composition variable associated with MES was lactose (r = 0.18; P < 0.01). Subclinical IMI did not affect MES when the comparison was performed using both models (1 and 2). Likewise, MES did not change when infected quarters were sorted into two groups of pathogens (major, minor and infrequent; and contagious, environmental, minor and infrequent) and compared with healthy mammary quarters. Considering the results of both models, subclinical IMI did not affect MES of dairy cows.

Phenotyping and genotyping of streptococci in bovine milk in Argentinean dairy herds

2010 ◽  
Vol 58 (3) ◽  
pp. 287-295 ◽  
Author(s):  
Elina Reinoso ◽  
Silvana Dieser ◽  
Luis Calvinho ◽  
Cristina Bogni ◽  
Liliana Odierno
Keyword(s):  
16S Rdna ◽  
Bovine Milk ◽  
Species Level ◽  
Dairy Herds ◽  
Polymorphism Analysis ◽  
Biochemical Tests ◽  
Milk Samples ◽  
Streptococcal Species ◽  
Milk Hygiene ◽  
Conventional Scheme

Most veterinary and milk hygiene laboratories identify streptococci and enterococci based on serological and biochemical tests. The analysis of 16S rDNA was suggested to be used for more exact identification; however, its use has not been considered so far in monitoring studies. The objective of the present study was to compare a conventional phenotypic method with restriction fragment length polymorphism analysis of 16S rDNA (16S rDNA RFLP) for identification of streptococci isolated from composite milk samples collected in connection with intramammary infection (IMI) in six Argentinean dairy farms. Composite milk samples (n = 1223) from cows belonging to six herds were collected for bacteriological analysis. Twelve reference strains and fifty streptococci or streptococcuslike isolates were identified to species level by the API 20 Strep system, conventional biochemical tests and 16S rDNA RFLP in a blind assay. The remaining streptococci or streptococcus-like isolates (n = 40) were identified to the species level both by 16S rDNA RFLP and conventional biochemical tests. As indicated by Kappa values, agreement between the 16S rDNA RFLP and the conventional scheme for identification ofStreptococcus agalactiae, S. dysgalactiae, S. uberis, S. equinusandEnterococcus faecaliswas 0.91, 0.73, 0.92, 0.81 and 0.85, respectively. Together with the less frequently isolated streptococcal species, the conventional scheme correctly identified 77 out of 90 isolates (85.5%). Thus, the use of 16S rDNA RFLP is considered valuable for monitoring studies due to its affordable cost for standard laboratories.

scholarly journals Antimicrobial Susceptibility of Streptococci Most Frequently Isolated from Czech Dairy Cows with Mastitis

2019 ◽  
Vol 19 (3) ◽  
pp. 679-694
Author(s):  
Soňa Šlosárková ◽  
Kateřina Nedbalcová ◽  
Jaroslav Bzdil ◽  
Petr Fleischer ◽  
Monika Zouharová ◽  
...  
Keyword(s):  
Antimicrobial Susceptibility ◽  
Susceptibility Testing ◽  
Subclinical Mastitis ◽  
Multidrug Resistant ◽  
Dairy Herds ◽  
Multiple Resistance ◽  
Milk Samples ◽  
Streptococcus Uberis ◽  
Amoxicillin Clavulanic Acid ◽  
Mastitis Pathogens

AbstractThe aim was to investigate the antimicrobial susceptibility of most frequently isolated streptococci from Czech dairy herds. A total of 3,719 quarter milk samples were collected and cultivated between January 2017 and June 2018 from cows with clinical or subclinical mastitis from 112 farms. Only one isolate of each species, collected from the same farm per six-month period, was included in the susceptibility testing. The susceptibilities of Streptococcus uberis (163 isolates) and S. dysgalactiae (25 isolates) to 10 antimicrobials (penicillin – PEN, amoxicillin/clavulanic acid – AMC, ceftiofur – EFT, clindamycin – CLI, gentamicin – GEN, streptomycin – STR, trimethoprim/sulfamethoxazole – SXT, enrofloxacin – ENR, tetracycline – TET, rifampicin – RIF) from 9 groups were determined by measuring their minimum inhibitory concentrations. The percentages of resistant S. uberis isolates to the antimicrobials were as follows: TET (63.2%), STR (52.1%), CLI (30.1%), and RIF (2.5%). Intermediate susceptibility was found to RIF (63.2%), PEN (35%), ENR (2.5%), EFT (1.8%), and AMC (1.2%). All the S. uberis isolates were susceptible to GEN and SXT (100%). However, only 6.7% of S. uberis isolates were susceptible to all tested antimicrobials, and 38.7% of isolates were multidrug resistant (≥ 3 groups of antimicrobials). All the S. dysgalactiae isolates were susceptible to PEN, AMC, EFT, GEN, SXT, and ENR (100%). Resistant S. dysgalactiae isolates were found to TET (60%), STR (28%), CLI (12%), and intermediate to TET (24%) and RIF (20%). Sixteen percent of S. dysgalactiae isolates were multidrug resistant. The relatively high occurrence of (multiple) resistance, relative to mastitis pathogens, highlights the importance of monitoring this condition in dairy herds.

scholarly journals Antimicrobial Study of Clinical Mastitic Milk Samples of Cows in and Around Bikaner

2019 ◽  
Vol 15 (02) ◽  
pp. 74-75
Author(s):  
Sandhya Morwal ◽  
AP Singh ◽  
SK Sharma ◽  
Deepika Goklaney
Keyword(s):  
Mammary Gland ◽  
Pathogenic Bacteria ◽  
Clinical Mastitis ◽  
Economic Losses ◽  
Dairy Herds ◽  
Cost Of Treatment ◽  
Milk Samples ◽  
Dairy Animals ◽  
Mastitic Milk ◽  
Major Alteration

Mastitis, an inflammatory condition of the mammary gland, is one of the most economically important diseases of dairy herds. This disease is more prevalent in high yielding dairy cattle. It is caused due to colonization of pathogenic bacteria causing series of events that lead to major alteration in the mammary tissues and composition of milk secreted from the tissue cells (Batavani et al., 2007; Goel et al., 2008). Clinical mastitis is recognized by abnormal milk, varying degree of mammary gland inflammation (redness, heat, swelling and pain) with or without illness of the cow. Mastitis in dairy animals leads to heavy economic losses due to reduction in milk production, altered composition, and cost of treatment of affected animals. The clinical mastitis in different cases can be identified by external symptoms such as udder swelling, touch, pain, redness and hardness of the udder. The objective of the present study was to assess the sensitivity of bacterial isolates from mastitic milk samples of cows to some selected antibiotics.

NGHIÊN CỨU SỰ NHẠY CẢM VỚI KHÁNG SINH CỦA MỘT SỐ CHỦNG VI KHUẨN GÂY VIÊM ĐƯỜNG HÔ HẤP CẤP Ở TRẺ EM DƯỚI 6 TUỔI TẠI BỆNH VIỆN NHI THANH HÓA 2009 - 2014

2020 ◽  
Vol 4 (3) ◽  
Author(s):  
Hoai Do Ngoc
Keyword(s):  
Antimicrobial Susceptibility ◽  
Susceptibility Testing ◽  
Antimicrobial Susceptibility Testing ◽  
High Rate ◽  
Isolation Rate

From 43.574 fluid nasopharynx speciments of  the chidren inpatient under six we isolated total 21.769 types bacteria with isolation rate : 49.95%. In which the highest isolation rate for H. influenza, S. pneumoniae and M. catarrhalis were 13,94%; 7,11%; 1,43% respectively. Antimicrobial susceptibility testing shown all the types of  for H. influenza, S. pneumoniae and M. catarrhalis good susses to Fosphomycine, S. pneumoniae and M. catarrhalis good susses to Imipenem, H. influenza good susses to Azithromycine, S. pneumoniae good susses to Penicilline and Piperacilline, M. catarrhalis good susses to Tobramycine and Ofloxacine. All of  H. influenza, S. pneumoniae and M. catarrhalis were reported resistance to Tri/Sulpha, Chloramphenicol, Erythromycine in high rate.

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