Relatedness of Staphylococcus aureus isolates from bovine mammary gland suffering from mastitis in a single herd

2000 ◽  
Vol 67 (3) ◽  
pp. 429-435 ◽  
Author(s):  
MICHAEL ZSCHÖCK ◽  
JÜRGEN SOMMERHÄUSER ◽  
HUGO CASTANEDA
Keyword(s):  
Staphylococcus Aureus ◽  
Protein A ◽  
Economic Loss ◽  
Epidemiological Studies ◽  
Resistance Pattern ◽  
Chromosomal Dna ◽  
Staph Aureus ◽  
Epidemiological Tool ◽  
Plasmid Profiling ◽  
Different Strains

Mastitis can be caused by a number of bacteria (Philpot & Pankey, 1975), among which Staphylococcus aureus is one of the most important, responsible for considerable economic loss to the dairy industry (Jasper et al. 1982). Carrier rates have been studied extensively in dairy herds suffering from mastitis (Davidson, 1961, 1963).Subtyping bacteria is an important epidemiological tool; for example, antimicrobial susceptibility patterns (antibiograms) have been used for typing Staph. aureus in human medicine (Gillespie et al. 1990). Staph. aureus of bovine origin can be divided into several categories by biotyping (Devriese, 1984). Phage typing has proved useful in differentiating mastitis strains (Mackie et al. 1987) and plasmid profiling has been valuable in epidemiological studies of bovine Staph. aureus (Baumgartner et al. 1984).During the last 5 years genomic fingerprinting of Staph. aureus became a powerful tool for epidemiological typing. Numerous techniques for comparison of staphylococcal isolates have been developed and are becoming important in investigations of strain origin, clonal relatedness and epidemiology.The aim of this study was to determine the epidemiology of Staph. aureus isolates from mammary glands of cows from a single herd using modern molecular typing techniques, and to assess whether the same strain constantly colonizes the udder or cows are repeatedly infected by different strains. We investigated 26 Staph. aureus isolates obtained from quarter milk samples from 16 cows. These isolates were characterized biochemically, by their antibiotic resistance pattern, by two polymerase chain reaction (PCR) methods on the basis of coagulase (Coa) gene and protein A (Spa) gene (X region) polymorphism and by macrorestriction analysis of chromosomal DNA using pulsed-field gel electrophoresis (PFGE).

Protein A gene polymorphism analysis in Staphylococcus aureus strains isolated from bovine subclinical mastitis

1999 ◽  
Vol 66 (3) ◽  
pp. 449-453 ◽  
Author(s):  
MARIA CRISTINA DALLA POZZA ◽  
ANTONIA RICCI ◽  
GADDO VICENZONI
Keyword(s):  
Staphylococcus Aureus ◽  
Gene Polymorphism ◽  
Protein A ◽  
Rflp Analysis ◽  
Epidemiological Studies ◽  
Molecular Techniques ◽  
Alternative Methods ◽  
Polymorphism Analysis ◽  
Modern Approach ◽  
Staph Aureus

Mammary infections caused by Staphylococcus aureus are still one of the most serious problems in dairy farms all over the world (Sischo et al. 1993), and the epidemiology of the infection has not yet been completely elucidated (Aarestrup et al. 1995). Any effective modern approach to this disease must therefore be based on more comprehensive epidemiological studies, conducted with valid microbiological typing tools.A technique for use in epidemiological studies should identify many types, and should be inexpensive, quick and easy to perform, but above all reproducible. Among the available methods, phage typing has up to now been widely and successfully used in differentiating strains of Staph. aureus isolated from cattle with mastitis (Mackie et al. 1987; Fox et al. 1991), but it has some limitations, being a technically demanding method subject to considerable experimental as well as biological variation (Maslow et al. 1993). Moreover, in some studies the number of strains that could not be typed with available bacteriophage panels has been high (Carroll & Francis, 1985; Farah et al. 1988).Alternative methods have been investigated, and of these molecular techniques have been the most intensely studied (Aarestrup et al. 1995). In studies of human infections caused by Staph. aureus, analysis of the so-called X region of protein A gene polymorphism has been a useful epidemiological marker (Frénay et al. 1994). This gene is ∼2150 bp and harbours some functionally distinct regions: an FC-binding region, the so-called X region and, at the C terminus, a sequence required for cell wall attachment (Guss et al. 1984; Frénay et al. 1994). The X region polymorphism depends on the presence, within the region itself, of a varying number of 24 bp repeats, highlighted by the amplification of this highly polymorphic DNA region and its subsequent restriction fragment length polymorphism (RFLP) analysis (Frénay et al. 1994).Human epidemic (MRSA) and non-epidemic methicillin-resistant Staph. aureus (non-MRSA) strains, which both cause infections but have completely different infection patterns, have been successfully distinguished by analysis of this polymorphism (Frénay et al. 1994). However, protein A has been identified in only 93% of Staph. aureus strains isolated from bovine intramammary infections (Poutrel & Ducelliez, 1979; Johne & Jarp, 1988).The aim of the present study was to determine whether the gene for protein A of Staph. aureus (Spa) was present in Staph. aureus strains isolated from cases of subclinical bovine mastitis. This was carried out using the polymerase chain reaction (PCR), as suggested by Frénay et al. (1994). In addition, we have investigated the genetic polymorphism related to the X region of the gene, by means of PCR amplification and subsequent RFLP analysis. Finally we verified the stability of this region after in vitro subculture.

scholarly journals To determine the resistance pattern of Staphylococcus aureus in pus samples

2021 ◽  
Vol 7 (4) ◽  
pp. 292-294
Author(s):  
Vasundhara Sharma ◽  
Versha Rajput ◽  
Umar Farooq ◽  
Sudhir Singh ◽  
Shweta R Sharma ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Antibiotic Sensitivity ◽  
Positive Culture ◽  
Blood Stream ◽  
Diffusion Method ◽  
Resistance Pattern ◽  
Staph Aureus ◽  
Care Workers ◽  
Accurate Treatment ◽  
Patient’S Outcome

Staphylococcus aureus is a common health problem occuring as an important nosocomial pathogen, causing urinary tract infection, surgical site, blood stream and soft tissue infection. The aim of this research was conducted to determine MRSA and VRSA from the pus samples of admitted patients.The aim and objective of study was to isolate the resistance pattern of Staphylococcus aureus in pus samples and their AST. A total of 158 positive culture Staph aureus were taken from pus samples for the study during December 2019 - October 2020. Samples were cultured on Blood and MacConkey agar then incubated at 37C for 24 hours. The modified Kirby Bauer's disc diffusion method was used to test antibiotic sensitivity of staphylococcus isolates. In total of 158 positive culture of Staphylococcus aureus, 66 (41.7%) were found to be MRSA and 4 (2.5%) were found to be VRSA. Out of 158 Staph aureus, 146 (92.4%) were resistant to Penicillin, followed by Amoxycillin 140 (88.6%), Ampicillin 139 (87.9%), Erythromycin 91 (57.5%), Cefoxitin 66 (41.7%), Gentamycin 56 (35.4%), Amikacin 52 (32.9%) and Teicoplanin 37 (23.4%).: An antibiotic policy and screening of susceptibility patterns of MRSA may help in reducing the prevalence rate of MRSA and antibiotic resistance. To stop its spread to the population, it is very important to eliminate MRSA colonization in patients and health care workers. Accurate treatment helps to reduce the rate of morbidity and improvement of patient’s outcome.

scholarly journals Plasmid analysis as an epidemiological tool in neurosurgical infections with coagulase-negative staphylococci

1987 ◽  
Vol 98 (3) ◽  
pp. 231-239 ◽  
Author(s):  
Åke Örtqvist ◽  
Ulrika Ransjö ◽  
Bengt Wretlind
Keyword(s):  
Antibiotic Resistance ◽  
Resistance Pattern ◽  
Enzyme Analysis ◽  
Restriction Enzyme Analysis ◽  
Coagulase Negative Staphylococci ◽  
Antibiotic Resistance Pattern ◽  
Plasmid Analysis ◽  
Epidemiological Tool ◽  
Resistance To Penicillin ◽  
Different Strains

SUMMARYCoagulase-negative staphylococci isolated from blood or spinal fluid during a period of 1 year in a department of neurosurgery, were analysed by biotyping, antibiotic resistance pattern and plasmid profiles. Altogether 41 isolates from 19 patients were studied. About 90% of the isolates wereStaphylococcus epidermidis. The antibiotic resistance pattern seemed to be closely related to antibiotic usage in the unit. Most common was resistance to penicillin (63%), trimethoprimsulphamethoxazole (49%) and cloxacillin (39%) while resistance to gentamicin was seen in only one strain.In several cases species and antibiograms were identical in isolates from different patients. Plasmid pattern analysis could then be used for identification of different strains. In one instance, plasmid pattern and restriction enzyme analysis confirmed that two patients probably were infected by the same strain.

scholarly journals Virulence factors and antimicrobial resistance of Staphylococcus aureus isolated from bovine mastitis in Rio de Janeiro

2009 ◽  
Vol 29 (5) ◽  
pp. 369-374 ◽  
Author(s):  
Shana M.O. Coelho ◽  
Elina Reinoso ◽  
Ingrid A. Pereira ◽  
Lidiane C. Soares ◽  
Mirta Demo ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Virulence Factors ◽  
Protein A ◽  
Bovine Mastitis ◽  
Subclinical Mastitis ◽  
Resistance Pattern ◽  
Igg Binding ◽  
Amplicon Size ◽  
Resistance To Penicillin ◽  
Single Amplicon

The study was conducted to characterize pheno-genotypically the virulence factors and resistance pattern of Staphylococcus aureus isolates from milk samples of cows with subclinical mastitis. All hemolytic isolates presented beta-hemolysin, and 38% of the non-hemolytic isolates were able to express hemolysins in the presence of a beta-hemolytic strain. The amplification of the coa-gene displayed four different size polymorphisms with about 400 bp, 600 bp, 700 bp and 900 bp. The spaA gene that encodes the IgG-binding region of protein A revealed sizes of 700 bp and 900 bp. The amplification of region X from spaA yielded a single amplicon for each isolate with the prevalent amplicon size being of 180 bp. Amplification of sae gene yielded an amplicon size of 920 bp in 71% of the isolates. Antibiotic resistance pattern revealed that 42% S. aureus were susceptible to all antimicrobials tested. Seven different antibiotic patterns were observed. Our results indicated that 47% and 25% of S. aureus strains exhibited resistance to penicillin and oxacillin respectively. All oxacillin-resistant isolates were mecA-positive.

scholarly journals SpaDiversity among MRSA and MSSA Strains ofStaphylococcus aureusin North of Iran

2010 ◽  
Vol 2010 ◽  
pp. 1-5 ◽  
Author(s):  
Fatemeh Shakeri ◽  
Abolfath Shojai ◽  
Masoud Golalipour ◽  
Somaye Rahimi Alang ◽  
Hamid Vaez ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Average Length ◽  
Protein A ◽  
Dual Band ◽  
Gene Variation ◽  
North Of Iran ◽  
Mrsa Strains ◽  
Different Strains ◽  
Tract Infections ◽  
Healthy Carriers

Protein A ofStaphylococcus aureusis a pathogenic factor whose encoding gene,spa, shows a variation in length in different strains. In this study thespagene variation inS. aureusisolated from healthy carriers and patients was studied, We also compared this variation among MRSA with MSSA strains. 208 strains ofStaphylococcus aureuswhich we were isolated from Gorgan, north of Iran were studied, 121 cases from patients and 87 cases from healthy carriers, 59 out of them wereMRSAand 149MSSA. Samples DNA were extracted and amplified by specific primer ofspagene. In 4 (3.8%) strains of them nospagene was detected, and 10.6% had a dual band (1200 and 1400 bp). In strains with one band, the length ofspagene differed from 1150 to 1500 bp. The most prevalent length was 1350–1400 bp (37%). The frequencies of shortspabands (1150–1200 bp) in patients strains were significantly higher. In 4 (3.8%) strains of them nospagene was detected, and 10.6% had a dual band (1200 and 1400 bp). In strains with one band, the length ofspagene differed from 1150 to 1500 bp. The most prevalent length was 1350–1400 bp (37%). The frequencies of shortspabands (1150–1200 bp) in patients strains were significantly higher. Thespagene length of 1350–1400 bp in MSSA was more than in MRSA strains (P<.05). The average length ofspain isolated strains from urinary tract infections was more than others. It is concluded that the length ofspagene depends either on resistance to Methicillin or the source ofS. aureusisolation.

Molecular epidemiology of Staphylococcus aureus in food samples based on the protein A gene polymorphic region DNA sequence

2010 ◽  
Vol 56 (1) ◽  
pp. 18-21 ◽  
Author(s):  
M. M. Soltan-Dallal ◽  
Z. Salehipour ◽  
Jalil F. Mehrabadi
Keyword(s):  
Staphylococcus Aureus ◽  
Protein A ◽  
Meat Products ◽  
Epidemiological Studies ◽  
Food Samples ◽  
Biochemical Tests ◽  
Polymerase Chain ◽  
D 20 ◽  
Spa Gene ◽  
Important Disease

Foodborne disease due to Staphylococcus aureus is a common and important disease worldwide. Molecular typing of S. aureus strains plays a crucial role in epidemiological studies examining the origin and performing surveillance of major infections. In this survey, we collected 913 food samples and detected 93 S. aureus isolates by using culture and biochemical tests. Subsequently, the X region of the protein A gene was amplified by the polymerase chain reaction (PCR) and amplicons were digested with HaeII and HindIII. Seven different patterns, ranging in length between 1200 and 1600 bp, showed gene polymorphism of the spa gene. The most prevalent spa types were D (20%) and C (16%) in dairy products and D (6%) and E (3%) in meat products. Consequently, 16 genotypes were obtained by HaeII digestion. Typeability of PCR – restriction fragment length polymorphism in genotyping of S. aureus strains in our study was perfect. Therefore, this method is a reliable, rapid, and powerful system in epidemiological investigations.

scholarly journals Rapid Detection of Epidemic Strains of Methicillin-Resistant Staphylococcus aureus

1999 ◽  
Vol 37 (3) ◽  
pp. 690-693 ◽  
Author(s):  
Thomas A. Wichelhaus ◽  
Sylvia Kern ◽  
Volker Schäfer ◽  
Volker Brade
Keyword(s):  
Staphylococcus Aureus ◽  
Rapid Detection ◽  
Methicillin Resistant Staphylococcus Aureus ◽  
Susceptibility Testing ◽  
Protein A ◽  
Surface Proteins ◽  
University Hospital ◽  
Chromosomal Dna ◽  
Methicillin Resistant ◽  
Specific Antibodies

Fifty methicillin-resistant Staphylococcus aureus(MRSA) initial isolates obtained from patients hospitalized in the orthopedic clinic of the Frankfurt University Hospital and 150 methicillin-sensitive Staphylococcus aureus (MSSA) isolates were investigated in this study to determine whether the Slidex Staph-Kit is capable of differentiating between MRSA and MSSA owing to its unique performance characteristics. The Slidex Staph-Kit is a combined latex hemagglutination test designed to detect clumping factor, protein A, and a specific surface immunogen for S. aureus. Clumping factor-positive strains cause erythrocytes sensitized with fibrinogen to hemagglutinate, thereby resulting in visible red clumps. S. aureus strains deficient in clumping factor agglutinate latex particles sensitized with specific antibodies against surface proteins of S. aureus, thereby resulting in visible white clumps. Our results demonstrate that white clumping has a 99% specificity as well as a 98% positive predictive value for MRSA. Clumping factor-negative MRSA, which have been reported to occur in several countries, are epidemic in the Frankfurt area and account for 80% of all MRSA initial isolates in the orthopedic clinic of the Frankfurt University Hospital. Genotyping of all MRSA isolates by macrorestriction analysis of chromosomal DNA revealed that 83% of clumping factor-negative MRSA are closely related to the “southern-German” epidemic strain. This is the first study demonstrating the Slidex Staph-Kit’s capability for identifying epidemic clumping factor-negative S. aureus strains as methicillin resistant even prior to antimicrobial susceptibility testing.

scholarly journals Rapid cost-effective subtyping of meticillin-resistant Staphylococcus aureus by denaturing HPLC

2006 ◽  
Vol 55 (8) ◽  
pp. 1053-1060 ◽  
Author(s):  
F. Jury ◽  
M. Al-Mahrous ◽  
M. Apostolou ◽  
S. Sandiford ◽  
A. Fox ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Protein A ◽  
Cost Effective ◽  
Epidemiological Studies ◽  
Heteroduplex Analysis ◽  
Reference Laboratory ◽  
Staphylococcal Protein A ◽  
Gene Variation ◽  
The Uk

The importance of meticillin-resistant Staphylococcus aureus (MRSA) in hospital-acquired infection is widely acknowledged. The UK government has stated that MRSA bloodstream infection rates will have to be halved by 2008. Such radical improvements will require advances on several fronts. Screening for MRSA in high-risk patients on arrival at hospital allows isolation of carriers and reduces transmission to staff and other patients. Concurrent subtyping of MRSA could also inform outbreak investigations and long-term epidemiological studies. The variability within the staphylococcal protein A, or spaA, gene-repeat region can be used as a marker of short- and long-term genetic variation. A novel application is described of denaturing HPLC (DHPLC) for rapid, inexpensive characterization of spaA gene amplification products, without the need for DNA sequence determination. The method allowed rapid and precise sizing of spaA gene-repeat regions from 99 S. aureus strains and was combined with heteroduplex analysis, using reference PCR products, to indicate the spa type of the test isolate. The method allowed subtyping of strains in less than 5 h from receipt of a primary isolation plate. When applied to an outbreak that occurred during this study, the authors were able to demonstrate relatedness of the isolates more than 5 days before results were received from a reference laboratory. If combined with direct amplification from swabs, DHPLC analysis of spaA gene variation could prove extremely valuable in outbreak investigation and MRSA surveillance.

Assessment of the Multifactorial Effect on Antimicrobial Sensitivity in Positive Staphylococcus Aureus Clinical Isolates from Assir Region, Saudi Arabia

2012 ◽  
Vol 13 (2) ◽  
pp. 152-159 ◽  
Author(s):  
Nazar M Abdalla ◽  
Waleed O Haimour ◽  
Amani A Osman ◽  
Hassan Abdul Aziz
Keyword(s):  
Staphylococcus Aureus ◽  
Saudi Arabia ◽  
Culture Media ◽  
Meca Gene ◽  
Laboratory Data ◽  
Age Groups ◽  
Clinical Isolates ◽  
Diabetic Patients ◽  
Staph Aureus ◽  
Antimicrobial Sensitivity

General objectives: This study aimed at assessment of factors affecting antimicrobial sensitivity in Staphylococcus aureus clinical isolates from Assir region, Saudi Arabia. Materials and Methods: In this study, eighty one patients presented with Staph. aureus infections either nosocomial or community acquired infections were involved by collecting nasal swabs from them at Aseer Central Hospital General Lab. These patients were from all age groups and from males and females during the period of Jan 2011- Jun 2011. These samples were undergone variable laboratory procedures mainly; bactech, culture media, antibiotics sensitivity test using diffusion disc test (MIC) and molecular (PCR) for detection of mec A gene. Clinical and laboratory data were recorded in special formats and analyzed by statistical computer program (SPSS). Results: Showed that; Descriptive and analytical statistical analysis were performed and final results were plotted in tables. In Staph aureus MecA gene positive cases (50) showed: Oxacillin/ Mithicillin, Ciprofloxacin and Fusidin resistant in diabetic patients were 13, 26.0%, 9, 18% and 7, 14% respectively and in non diabetic patients were 37, 74.0%, 22, 44% and 20, 40% respectively. While no sensitivity in diabetic and non diabetic patients using Oxacillin/ Mithicillin. In Staph aureus MecA gene negative cases (31) showed: Oxacillin/ Mithicillin, sensitivity in diabetic patients (5, 16.1%) and in non diabetic were (26, 83.9%). While no resistant in diabetic and non diabetic patients. In Ciprofloxacin and Fusidin resistant in diabetic patients were 1, 3.2% and 1, 3.2% respectively and in non diabetic patients were 12, 38.7% and 7, 22.6%respectively. Erythromycin in Staph aureus ( MecA gene) positive cases (50) showed: resistant in age (0-15) years were (5, 10%), (16-50) years were (16, 32%) and ( ›50 years) were (12, 24%). Erythromycin in Staph aureus (MecA gene) negative cases (31) showed: resistant in age (0-15) years were (6, 19.3%), (16-50) years were (5, 16.1%) and ( ›50 years) were (3, 9.7%). Conclusion: Drugs resistance is a major progressive multifactorial problem facing the treatment of Staph aureus infections. DOI: http://dx.doi.org/10.3329/jom.v13i2.12750 J Medicine 2012; 13 : 152-159

Sign in / Sign up

Export Citation Format

Share Document