Coagulase Enzymes Research Articles

Screening and genome analysis of potential probiotic lactic acid bacteria with broad-spectrum antibacterial activity from Sichuan sun-dried vinegar grains (Cupei)

This study aimed to isolate lactic acid bacteria (LAB) with broad-spectrum antibacterial activity and potential probiotic properties from the Sichuan sun-dried vinegar grains (Cupei), a traditional Chinese bran fermented vinegar. A total of 115 LAB strains were isolated from the Cupei, among which 33 inhibited both Gram-negative and Gram-positive indicator bacteria. Subsequently, 27 strains devoid of hemolytic and coagulase enzyme activity were selected for further evaluation of their probiotic properties and Lactobacillus plantarum SC27 was revealed as a potential probiotic strain, as it demonstrated notable gastrointestinal tolerance, hydrophobicity, auto-aggregation ability, and multiple antibiotic sensitivity, and particularly noteworthy for broad-spectrum antibacterial activity. Whole genome sequencing further unveiled that SC27 harbored bacteriocin gene clusters likely responsible for bacteriocin production, as well as an extensive variety of phosphotransferase system transporters facilitating its adaptability to diverse food substrates. In addition, based on the analysis of genes involved in antibiotic resistance, virulence factor, we speculated that SC27 is safe for application in food industry. Furthermore, the genomic analysis revealed several stress-response and adhesion-related genes, known to confer beneficial properties to probiotic bacteria. In conclusion, our comprehensive genomic and phenotypic analyses confirm the potential of SC27, with broad-spectrum antibacterial activity, as a probiotic used in food.

Recovery of Methicillin Resistant Staphylococci from Public Vehicles in Kathmandu

Objectives: The purpose of this study was to determine the extent of staphylococcal contamination in public vehicles in Kathmandu and to determine the antibiotic susceptibility pattern of the isolates. Methods: This cross-sectional study was conducted from December 2022 to May 2023 in the Microbiological laboratory of Padma Kanya Multiple Campus, Kathmandu. A total of 120 swab (sterile cotton swab moistened with sterile normal saline) samples were collected by gently rubbing the surfaces from 19 different micro-buses during the time of heavy passenger flow (office time) and transported to the laboratory maintaining a cold chain. Staphylococcus aureus and Coagulase negative Staphylococci (CoNS) were identified by their characteristic yellow pigmented colonies on Mannitol Salt Agar (MSA) and the ability of S. aureus to produce coagulase enzyme. Kirby-Bauer disc diffusion method was employed to test the antibiotic susceptibility pattern of isolates. MRSA were identified on the basis of the zone of inhibition (<21 mm diameter) against Cefoxitin 30mcg disc. Results: Out of 41 Staphylococcal isolates, 46.36% were S. aureus, among which 84.21% exhibited methicillin resistance (MRSA). All Coagulase Negative Staphylococci (CoNS) isolates (n=22) were resistant to methicillin (MRCoNS). MRSA and MRCoNS isolates were also resistant to antibiotics like Penicillin and Erythromycin. Conclusion: This study reports relatively high occurrence of MRSA in environmental samples, predominantly in areas frequently touched by people indicating the possibility of easy spread of these organisms and subsequent opportunistic infections.

Biochemical typing and evaluation of pathogenicity in vulvovaginal isolates of Candida albicans complex

Candida albicans, C. dubliniensis, and C. africana form the Candida albicans complex. To identify the phenotypic and pathogenic characteristics of isolates of the C. albicans complex preserved in a collection. Three hundred presumptive strains of the C. albicans complex were evaluated using CHROMagarTM Candida. Germ tube production was determined by three methods, chlamydospores formation was assessed and colonies were characterized in artisanal agars (Rosmarinus officinalis and Nicotiana tabacum). MALDI-TOF was used as the gold standard identification test. To detect pathogenicity factors, we evaluated the hemolytic activity of each isolate and cocultured with Staphylococcus aureus, coagulase enzyme production, and biofilm formation. Out of the 300 isolates, 43.7% produced germ tube in the heart-brain infusion broth and 47% of the isolates produced chlamydospores. In the artisan media, 6% of the isolates produced brown colonies on rosemary agar and 5% did so on tobacco agar. None of the strains hemolyzed the blood agar alone or cocultured with S. aureus. However, 50% of the isolates hemolyzed the potato dextrose agar supplemented with blood. All strains were coagulase producers, and biofilm production was variable. For germ tube production, the human serum method showed the same positivity as the milk broth method. All isolates were identified as C. albicans by MALDI-TOF. The use of proteomics, molecular tests or a combination of methods is required for species identification.

Identification of bacterial morphology and catalase coagulation test on propionibacterium acnes bacteria

Infection is a way for microorganisms such as bacteria, viruses and fungi to enter the skin which can cause skin problems such as acne. purpose of this study was to determine the morphology of p. acnes by gram staining method, then the catalase test was used to determine the ability of microorganisms to decompose hydrogen peroxide by producing catalase enzymes and coagulase test is a bacterial examination to detect the formation of coagulase enzymes that bind to the bacterial cell wall. The results obtained were that the morphology of the bacteria was in the form of salt and purple in color, the catalase test was positive and the coagulase test was positive. The conclusion obtained was that P. acnes ATCC 11827 was included in the group of gram-positive bacteria based on the results of gram staining and catalase and coagulase tests.

Detection of Virulence Factors from Candida spp. Isolated from Oral and Vaginal Candidiasis in Iraqi Patients.

Yeast-like fungi (YLF) of the genus Candida are unicellular microorganisms of relatively large size and rounded shape, aerobes, and belong to conditionally pathogenic microorganisms. The genus Candida includes approximately 150 species, which are classified as Deuteromycetes due to the absence of a sexual stage of development. This study aimed to identify virulence factors from Candida spp. isolated from oral and vaginal candidiasis. Fifty-eight oral and vaginal swab specimens were collected from patients, including (28) oral swabs from children and (30) vaginal swabs from different infected women. All isolates were subjected to direct examination, Morphological tests, Germ tube formation, growth at 45ºC, CHROM agar Candida culture, and VITEK 2 Compact system to ensure this diagnosis. (31) isolates were identified as Candida spp., including (21) (C. albicans (14), C. glabrata (1), C. guilliermondii (2), C. dubliniensis (3), C. parapsilosis (1)) were isolated from oral swabs and (10) isolates includedC. parapsilosis (4), C. albicans (6) were isolated from vaginal swabs. Moreover, these isolates had been detected to have some virulence factors, including phospholipase, esterase, proteinase, coagulase, hemolysin, and biofilm formation. Different species of Candida were isolated and identified from oral and vaginal. Phospholipase (Pz), Esterase (Ez), and Proteinase (Prz) were produced by 19 (61.29%), 16 (51.61%), and 26(83.87%), respectively, out of 31 isolates, whereas. All isolates produce coagulase enzyme except C. dubliniensis, which did not produce coagulase enzyme. All Candida spp. isolates produce hemolysin and biofilm formation in different percentages.

Molecular analysis of Staphylococcus aureus isolated from clinical samples and natural flora.

A total of 100 samples collected from the wound, abscess skin, and normal human flora were investigated for S. aureus identification. Overall, in 40 samples, S. aureus isolates were present, out of which most strains were isolated from normal human flora (50.0%), followed by wound (37.5%) and burn (12.5%) samples. Moreover, S. aureus isolates from all samples could produce extracellular enzymes (catalase, coagulase, urease, and hemolysin-ß) as virulence factors except for some isolates from normal flora samples (unable to produce coagulase enzymes). Therefore, genes encoding the enzymes coagulase and hemolysin were evaluated in 20 S. aureus isolates by PCR-specialized primers targeting co-specific genes. The PCR analysis revealed that clinical isolates included both genes. Contrarily, 6 isolates of the normal flora lacked the coa gene, revealing bacterial fingerprints that can be used to distinguish between isolated bacteria and human beings.

Microbiological hazard identification in selective food products and their association with food safety practices in Hyderabad, India

Microbiological risk assessment (MRA) is an emerging tool for evaluating the safety of food and water supplies. In this study, identification of microbial hazards in selected samples and their association with food safety practices were seen. Data were analyzed using Analysis of Variance (ANOVA) through the General Linear Models (GLM) procedure of the statistical analysis system software (SPSS version-11.5, 2003). The least significant differences were used to test means at p<0.05. Odds ratio (OR) values with 95% CI were computed to obtain the risk of the presence of the foodborne pathogen in a particular food. Analysis of 600 food samples indicated that (50%) was high in raw chicken samples followed by carrot salad (41%). The other emerging foodborne pathogens like (MRSA), , and were not detected in any of the food samples. Pathogens like (73.5%) in khoa, (45%) and (62.7%) in paneer were detected. A total of (n=143) cultures were analyzed for enterotoxin and coagulase enzyme. Nine cultures (6.3%) showed a positive result for enterotoxin production. For the risk assessment of contamination in foods, coagulase test and toxin production of isolates have to be evaluated. A significant association was found between the type of storage and log concentration of in khoa, whereas, with water for washing hands, the status of nails, and cleaning cloth were contributing to foodborne pathogens in other products. There is a need to provide food safety training to food handlers to improve food safety.

Study the Factors affecting the Production of Coagulase Enzyme from Clinical Bacteria Isolated

This study included isolation and diagnosis of Coagulase Positive Staphylococci bacteria fromclinical samples of (21) samples from the Teaching Hospital for Women and Children in Ramadi andincluded nasal swabs, Ear swabs, Normel skin swabs, Wound swabs, Urine Swabs and Abscesses.These isolates were diagnosed according to phenotypic and biochemical tests and vatic. After diagnosingthe developing colonies, it was found the Gram-positive bacteria formed a percentage (71.42%) of theclinical samples out of the total isolates that gave growth on the culture media, and the Staphylococcipositive for the plasma coagulase enzyme (Coagulase Positive Staphylococci) was found. (71.42%) ofthe total Staphylococci bacteria obtained during isolation and diagnosis of clinical samples and a typeof Staphylococcus positive for coagulase (CPS), Staphylococcus. aureus, was diagnosed based on theSlide and Tube Coagulase Test.The sensitivity of CPS-positive staphylococcus to the antibiotic against the bacteria was tested and itwas found that the most effective antibiotic against S. aureus bacteria was Azithromycin as well as theantibiotic Gentamycin, as indicated by S. aureus has demonstrated resistance to Erythromycin as wellas Ciprofloxacin.The different conditions for the production of the plasma coagulase enzyme (Coagulase) for the bacterialisolate of local number (5) were studied, and it was noticed that there is a great similarity in theseconditions except for some differences. The study and it was noticed that the appropriate temperature inthe production of the enzyme for the local bacterial isolate number (5) is the temperature (35) C, and itwas noticed that the pH that led to the increase in the production of the enzyme is (PH10).After (24hr)incubator , and the results were consistent in terms of good production of the enzyme in the mentionedisolation when using a incubator shaker at a speed (100) .

Identifying possible target of action of 4,4a-dihydroxanthones in bacterial cells

Introduction. Partially hydrogenated derivatives of xanthone, dihydroxanthones, are being intensively studied. They are of interest due to their antimicrobial, antitumor, and antioxidant effects. Many researches are focused on the study of the cytotoxicity of dihydroxanthones and very little information is available on their antimicrobial activity. Therefore, the study of the antimicrobial activity and mechanism of action of new synthetic derivatives of 4,4a-dihydroxanthone is relevant. Preliminary studies have demonstrated that 4,4a-dihydroxanthones are active against gram-positive bacteria and have a pronounced anti-staphylococcal effect. Namely, 5-bromo-4,4-dimethyl-7-chloro-4,4a-dihydroxanthone (BDC-DX) was shown to be the most active derivative.Aim of the study was to determine the possible target of action of the active derivative of BDC-DX in bacterial cells and its acute toxicity.Materials and methods. The method of measuring the intensity of absorption of the crystal violet dye by bacteria cells was used to prove the effect of BDC-DX on the permeability of the cytoplasmic membrane in bacterial cells. The plasma coagulase activity of Staphylococcus aureus was tested under the action of dihydroxanthone to determine the effect of dihydroxanthone on the process of protein synthesis. Plasmid DNA digestion method was used to study the effect of the compound on bacterial DNA. The acute toxicity of BDC-DX was determined by the express method of V.B. Prozorovsky.Results and discussion. BDC-DX increased the permeability of the cytoplasmic membrane of S. aureus. Dihydroxanthone did not directly affect the plasma coagulase activity of Staphylococcus and showed a weak damaging effect on bacterial DNA. The compound induced breaks in plasmid DNA at a very high concentration — 1 mM or 384 pg/ml and higher. BDC-DX is a low-toxic compound (the average lethal dose for oral administration of the compound is 1710 ± 170 mg/kg, the average lethal dose for intraperitoneal administration of the compound is 116.9 ± 13.3 mg/kg).Conclusion. For the first time, in-depth study of the possible mechanism of action of a new synthetic biologically active compound from the group of 4,4a- dihydroxanthones, BDC-DX, was conducted. A likely target of 5-bromo-4,4-dimethyl-7-chloro-4,4a-dihydroxanthone in S. aureus cells is the cytoplasmic membrane. BDC-DX did not affect the process of protein synthesis, namely the activity of the plasma coagulase enzyme. The compound had no pronounced damaging effect on bacterial DNA. It was found that 4,4a-dihydroxanthone refers to low-toxic compounds.

Non-Candida albicans Candida species: virulence factors and species identification in India.

Background and Purpose: The predominant cause of candidiasis was Candida albicans which has recently changed to non-Candida albicans Candida (NCAC) (i.e., Candida spp. other than the C. albicans). The NCAC spp., earlier considered non-pathogenic or minimally virulent, are now considered a primary cause of morbidity and mortality in immunocompromised individuals. Given the NCAC spp.has become more common in clinical cases, this study aimed to determine the prevalence of NCAC spp. in different clinical specimens and assess a few of their virulence factors.Materials and Methods: Routine samples for bacterial culture and sensitivity that showed colony characteristics, like Candida on Blood Agar and microscopic features resembling Candida spp., were processed further. Candida isolates underwent tests for chlamydospore formation and biochemical tests, including sugar fermentation and sugar assimilation tests. These were grown at 42oC, and their colony color was identified using HiCromeTM Candida Differential Agar (HiMedia Laboratories Pvt. Ltd., Mumbai, India), HiCandidaTM Identification Kit (HiMedia Laboratories Pvt. Ltd., Mumbai, India), and VITEK-2® Compact (Biomérieux, France). Virulence factors, such as adherence to buccal epithelial cells (ABEC), biofilm formation, hemolytic activity, and production of coagulase enzyme were also tested.Results: Mean age of the patients was 38.46 years with a male-female ratio of 1.36:1. In total, 137 Candida isolates were recovered; 45.3%, 19.7%, and 13.9% of the isolates were isolated from urine, vaginal swabs, and oropharyngeal swabs, respectively. Moreover, 55 (40.1%) isolates were those of C. albicans and 82 (59.9%) isolates belonged to NCAC spp., with C. tropicalis (23.4%) contributing highest among NCAC species. Furthermore, C. albicans (3; 50%) was the most common spp. in cases of candidemia. Haemolysin production (85.5%) and ABEC (78.2%) were the major virulence factors in C. albicans. C. tropicalis (59.4%) and C. dubliniensis (50%) showed maximum ABEC. Biofilm forming capacity was higher in C. tropicalis (78.1%) than C. albicans (67%).Conclusion: Results of this study suggest varied prevalence and virulence based on geographical locations, even within a subcontinent. It clearly indicates the emergence of the NCAC spp. and their predominance in different body fluids. Identification of Candida to the spp. level should become a routine in all laboratories.

Isolation and Genetic Identification of Endophytic Lactic Acid Bacteria From the Amazonian Açai Fruits: Probiotics Features of Selected Strains and Their Potential to Inhibit Pathogens.

The açai palm (Euterpe oleracea) is native to the Amazon basin, a humid tropical forest. High levels of total mesophilic bacteria with high diversity have been consistently reported in açai fruits. As local consumers have few digestive problems, the results of the present study reveal the lactic acid bacteria (LAB) recovered from açai fruits with characteristics that suggest they are possible candidates for probiotics and antagonistic potential against pathogens for the first time. Açai fruits were sampled from five different locations in the Eastern Amazonia floodplains. Sixty-six isolates were recovered from fruits and tested for some probiotic characteristics following FAO/WHO guidelines. Approximately 65% of the isolates showed no catalase or oxidase activity, Gram-positive staining or cocci and bacilli cell morphology. Furthermore, 48% of the isolates demonstrated preliminary characteristics that suggest safety for use, as they presented no coagulase enzyme activity or gamma-hemolysis. These strains were identified as belonging to the genera Lactiplantibacillus and Pediococcus, and 32 strains also presented resistance to vancomycin, ciprofloxacin and streptomycin. In addition, 28 isolates showed a survival rate, expressed as log cycle reduction, higher than 0.9 under gastric conditions (pH 2). All strains tested positive in bile salts deconjugation tests and showed a survival rate higher than 0.8 in the presence of this salt. Regarding antimicrobial activity against pathogens, all strains were able to inhibit Salmonella Typhimurium (ATCC® 14028TM) and 97% were capable of inhibiting Escherichia coli (ATCC® 25922TM). Concerning the results of in vitro antagonistic assays, three isolates (B125, B135, and Z183 strains) were selected for antagonistic tests using açai juice contaminated with these two pathogens. All tested LAB strains were able to inhibit pathogen growth in açai juice. In summary, açai fruits are a potential source of LAB isolates to be investigated as probiotics.

Molecular Identification and Virulence Factors Determination in Candida Species Isolated from Egyptian Patients

Candida-related infections are becoming a universal threat to the health of human who undergo immunosuppressive therapy or aggressive medical intervention. Objectives: The aim was to study the distribution of Candida species among winter and summer seasons and to determine the expression of their virulence factors. Methods: A total of 164 Candida isolates were collected from clinical specimens at Mansoura University Hospitals. Candida species were identified by polymerase chain reaction (PCR). Extracellular phospholipase, secretory aspartyl proteinase (SAP) and coagulase enzymes and biofilm formation were determined. SAP 9 and 10 genes were detected by PCR. Results: Non-albicans (NAC) isolates were more dominant than C. albicans isolates (P value C. tropicalis was the most prevalent (59.2%) followed by C. albicans (31.1%), then C. glabrata, C. krusie, unidentified NAC and C. kefyr in 3.7%, 2.4%, 2.4% and 1.2% respectively. Extracellular phospholipase activity was detected in 31.7% of Candida isolates. All C. albicans had phospholipase activity (100%) and one isolate of C. tropicalis was positive while other species were negative. SAPs activities were determined in 61.6% of Candida isolates and were detected in 70.1% and 62.7% among C. tropicalis and C. albicans isolates respectively. SAP9 and SAP 10 genes were detected in 27.7% and 12.9% of Candida isolates showed positive SAPs activity respectively and they were all C. albicans strains. Other species did not harbor either SAP9 or SAP10. Coagulase activity was detected in 80.4% of Candida isolates with higher activity in C. albicans (88.2%), followed by C. tropicalis (81.4%), then other NAC isolates. Biofilm formation was determined in 69.5% of Candida isolates and was more prevalent in C. tropicalis (82.5%) followed by C. albicans (19.6%), C. krusie (100%), unidentified NAC (75%), C. glabrata (33.3%) and C. kefyr (50%). Conclusion: NAC with a preponderance of C. tropicalis was the most common isolated Candida species. Biofilm production, proteinase, phospholipases and coagulase enzymes were observed in both C. albicans and NAC. SAP9 and SAP 10 genes were detected only in C. albicans strains.

Antimicrobial, physico-chemical properties of nitrogen-containing preparations of derivates of menthol, quinoline and phenol

Successful research by scientists of new synthetic substances of various chemical groups contributes to the broadening of the arsenal of antimicrobial drugs for the prevention and treatment of purulent-inflammatory diseases. Antimicrobial drugs, as a rule, suppress pathogenic, invasive, adhesive properties and reduce the resistance of microorganisms to antibiotics in pathogens of supportive inflammatory diseases; significantly increase the effectiveness of treatment of diseases of infectious origin. The purpose of the study was to study the physicochemical, antimicrobial properties of derivatives of menthol, phenol and quinoline. The results of the study of physicochemical, antimicrobial properties of six chemical compounds of menthol, quinoline, and phenol derivatives using the principle of complex research, in which physicochemical, microbiological methods were used, are presented. There was shown that quaternary ammonium compounds of the menthol derivatives were alike white powders with a molecular weight of 581–693, a melting point of 990 to 1850° C. The chemicals are soluble in water, ethanol. Quinoline preparations have a molecular weight of 687; 756, melting point 178–2000°C; dissolved in ethanol. Compounds of phenol had a molecular weight of 111, 112, a melting point of 1020, 1100°C was soluble in ethanol. It has been established that synthesized substances possess a wide spectrum of antimicrobial action on Gram-positive, Gram-negative bacteria, Candida albicans. In antibiotic resistant strains of Staphylococci no markers of resistance to drugs containing in the molecule menthol, phenol, quinoline were found. In complex physical and chemical systems, it was important to study the coefficient of surface tension of solutions of drugs, which was an important objective physical indicator of the molecular state of various drugs. Distilled water was used as a control. Experiments were performed according to a well-known technique. According to the results of the study, in the control the surface tension of water was it was found to be 55,70 dn/cm2. In an experiment with 0,1% solution of decamethoxin; the drug number 2 was 40,80 dn/cm2 and 38,20 dn/cm2. In derivatives of quinoline (DN, drug № 4), was 39,60 dn/cm2 and 34,50 dn/cm2. Solutions of phenol (preparations №5; №6) were characterized by surface tension 32,40–43,50 dn/cm2. Surface tension of solutions of preparations depended on their chemical structure. The antimicrobial properties of the preparations were determined on the museum and clinical strains of microorganisms, which had typical tynctorial, morphological, and cultural characteristics. For a complete biological characterization in strains of Staphylococci, the formation of coagulase enzymes, lecithovitellase, hemolysins, and mannitol fermentation in anaerobic conditions were studied. At 12 museum and clinical strains of bacteria, bacteriostatic and bactericidal effects of six drugs, which are derivatives of menthol (DK, №2), quinoline (DN, №4), phenol (preparations №5, №6), have been detected. Derivatives of menthol acted bactericidal to Staphylococci at doses of 0,48-3,9 μg/ml; Quinoline derivatives in the range of 7,8–15,6 μg/ml; derivatives of phenol 31,25–62,5 μg/ml, respectively. Staphylococci were highly resistant to phenol derivatives (31,25–62,5 μg/ml). Gram-negative bacteria exhibited high resistance to quinoline and phenol derivatives (250–500 μg/ml). Summing up the results of determining the antimicrobial action of antiseptics derivatives of menthol, quinoline, it should be emphasized that the drugs have high activity in relation to Staphylococci (0,24–7,8 μg/ml). Phenol derivatives have low bacteriostatic and bactericidal effects on Gram-negative bacteria (125–500 μg/ml), which limits their use in medicine.

Investigation of coa Gene Polymorphism in Clinical Isolates of Staphylococcus aureus in North of Iran

Staphylococcus aureus is a common pathogen potentially able to cause a wide range of infectious diseases in human and animals and coagulase enzyme is one of the important virulence factors of this bacterium. Polymorphism of the coagulase encoding gene (coa) is one of the molecular-based typing methods of S. aureus isolates. In this study, the polymorphism of the coagulase gene among MRSA and MSSA isolates were investigated using PCR-RFLP analysis. To perform coagulase gene typing, the repeated units encoding hypervariable regions of coagulase gene of 30 clinical isolates of S. aureus were amplified by the PCR technique; this was followed by AluI restriction enzyme digestion and analysis of restriction fragment length polymorphism (RFLP) patterns. In total two amplicons (680 bp and 750 bp) and four distinct RFLP banding patterns (280+400, 340+340, 280+470, and no digested amplicon of 750 bp) were observed. Genotype with PCR-RFLP patterns of 280+400 bp was predominated. The results indicated polymorphism in the investigated regions of coagulase gene. This polymorphism can be used for identification of S. aureus isolates and showing the epidemiological relationship among them.

Relationship between Antibiotic Resistance Patterns and Coagulase in Clinical Isolates of Staphylococcus aureus in Nowshahr and Chalous

Background: Staphylococcus aureus is one of the main agents of nosocomial infections. Identifying coagulase enzyme in staphylococcal infections is of great importance as it is one of the main virulence agents. The present study aims to accurately detect the presence of coagulase gene and its relationship with antibiotic resistance patterns in clinical isolates. Methods: In this experimental study, a total of 20 clinical samples of S. aureus and 10 clinical samples of coagulase-negative staphylococci were collected and identified from hospitals and laboratories in Nowshahr and Chalous. Antibiotic resistance patterns of isolates were examined based on CLSI and finally polymerase chain reaction was used in order to detect the presence of coagulase gene with specific primers. Treatments were examined in three intervals and the results were analyzed using one-way analysis of variance (ANOVA), univariate ANOVA, and t-test at the probability level of P < 0.05. Moreover, the comparison of means was presented using Duncan’s multiple range tests (P < 0.05). All the statistical analyses were carried out using SPSS (V. 16). Results: Suggested that isolates showed phenotypic presence of coagulase enzyme and had this gene genotypically as well. Coagulase-positive samples demonstrated higher resistance to tetracycline 20%, oxacillin 55%, gentamicin 20%, and kanamycin 25%. Statistically, there was no significant difference among inhibition zone diameters of oxacillin, nalidixic acid, gentamicin, kanamycin, and penicillin. Conclusions: Coagulase gene causes antibiotic resistance and coagulase-negative staphylococci are spreading rapidly and their resistance is increasing.

Coagulase gene polymorphism and antimicrobial susceptibility of Staphylococcus aureus isolated from bovine subclinical mastitis milk in Sidi-Bel-Abbes, Algeria

The study was conducted to identify and characterize Staphylococcus aureus in raw milk derived from subclinical mastitis in Sidi-Bel-Abbes Algeria. In this paper, we explore the possibility of detection of the coagulase gene (coa), which encodes the coagulase enzyme, by PCR analysis in antibiotic-resistant isolates, with the latex agglutination phenotype and free coagulase.Out of 336 samples of raw milk examined with California mastitis test (CMT) posi-tive; a total of 142 samples were bacteriologically positive with 56.34% Staphylococcus isolates, 21 (26.25%) isolates were confirmed as S.aureus. Nineteen (90.48%) isolates of S.aureus showed free coagulase on the tube agglutination test. Two atypical S.aureus strains (9.52%) were defective for the clumping factor and / or protein A , determined with the Staphytect plus test and the tube coagulase test. The isolates of S.aureus were resistant to penicillin and tetracycline with 76.19%. Two isolates (9.52%) of S.aureus re-sistant to meticillin (MRSA) were detected in this study, with a MIC of ≥4 μg / liter and a cefoxitin screen test with a MIC of ≥8 μg / liter, and 13 (61.9%) isolates were with a multiresistance phenotype. The 21 isolates were sub-jected to PCR amplification of the 3' end of the coa gene, 18 (85.71%) were revealed on a 1% agarose gel with a single band between 547 bp and 875 bp. The use of the PCR genotypic test to identify the profile of the coa gene can be used as an appropriate identification criterion for differentiating coagulases from S.aureus and for understanding their epidemiology.

Occurrence of Bovine and Human Methicillin-Resistant Staphylococcus aureus in Organised Dairy Farms of Andhra Pradesh, India

The present study was undertaken for isolation and molecular characterization of methicillin-resistant Staphylococcus aureus (MRSA) from nasal swabs of apparently healthy cattle, buffaloes and their handlers. Out of 513 nasal swab samples analyzed, 162 (31.6%) S. aureus isolates were recovered, of which 104 (20.2%) were found positive for coagulase enzyme production. On MeReSa HICHROM agar supplemented with cefoxitin, bluish green colonies were observed in 100% coagulase positive S. aureus isolates of livestock handlers followed by buffaloes (48.6%) and cattle (24%). Disc diffusion test showed resistance to both oxacillin and cefoxitin in 7 (77.7%), 16 (43.7%) and 20 (33.3%) coagulase positive isolates of livestock handlers, buffaloes and cattle, respectively. Both mecA and blaZ genes were detected in 3 (33.3%), 10 (28.6%) and 11 (18.6%) coagulase positive isolates of livestock handlers, buffaloes and cattle, respectively. Correlation between phenotypic resistance to oxacillin, cefoxitin and presence of mecA gene was not observed.

Molecular analysis of the coagulase gene in clinical and nasal carrier isolates of methicillin-resistant Staphylococcus aureus by restriction fragment length polymorphism.

The coagulase enzyme, encoded by the coa gene, is an important virulence factor of Staphylococcus aureus and can be used for typing of S. aureus isolates. In this study, coa gene typing was used to study the epidemiology of S. aureus at a university hospital in Hamadān (Iran). A total of 200 S. aureus strains were analysed, among which 150 were isolated from clinical samples and 50 were from nasal swab specimens of carriers. Methicillin resistance was confirmed by presence of the mecA gene by PCR. For polymorphism analysis, the coa gene was amplified by PCR and the products were subjected to restriction digestion using the enzyme AluI. Amplification of the coa gene produced five classes of bands based on size, ranging from 300bp to 800bp. The 600-bp amplicon included coa 3 genotype predominated in S. aureus isolated from clinical and carrier specimens (150/200; 75.0%). AluI digestion of the PCR products produced eight distinct restriction fragment length polymorphism (RFLP) patterns, designated coa 1-8. The results showed that the 700-bp and 800-bp amplicons formed two (coa 4a and 4b) and three (coa 5a, 5b and 5c) patterns following AluI digestion, respectively, whereas the 300-, 500- and 600-bp fragments generated unique patterns designated coa 1, coa 2 and coa 3, respectively. This study performed coagulase typing, a technique used to determine the molecular epidemiology of S. aureus clinical isolates. coa gene amplification has been considered a simple and accurate method for typing of S. aureus.

Antimicrobial Susceptibility and Coagulase Typing of MRSA Strains at Mymensingh Medical College

Antimicrobial susceptibility and coagulase typing of Staphylococcus aureus isolates, with particular emphasis to Methicillin Resistant S. aureus (MRSA), were studied among strains isolated from various types of specimens collected at Mymensingh. A newly developed panel of anti-sera against different coagulase enzymes was used for coagulase typing. The study included 79 strains of S. aureus and of those, 40 were identified as MRSA on the basis of resistance to oxacillin (1 gm) discs. The rate of resistance of S. aureus to different antibiotics observed: penicillin (88.61%), oxacillin (48.10%), cephradine (44.30%), gentamicin (27.85%), erythromycin (34.18%), fusidic acid (25.0%) and rifampicin (20.0%). None of the isolates showed vancomycin resistance. Both MRSA and non-MRSA strains were found belonging to Coagulase type VI. Drug resistance pattern and coagulase type of MRSA strains indicated a homogenous epidemiologic origin that needs further exploration including strains from different geographic locations.Bangladesh J Med Microbiol 2007; 01 (02): 56-60

Muscoid Diptera as potential vectors of bacterial agents on dairy farms in the northern region of Paraná, Brazil

The aim of this study was to determine the isolation frequency and microbial sensitivity profiles of Escherichia coli, Salmonella spp., and Staphylococcus spp. isolated from synanthropic flies captured in entomological traps placed near the milking parlors of 30 dairy farms located in Northern Parana, Brazil. In total, 192 flies were captured, and the isolation frequencies of muscids (21/30 = 70%) and califorids (27/30 = 90%) were significantly higher than that of sarcophagids (7/30 = 23.3%). Bacteriological tests on the internal contents and external surfaces of the flies showed that E. coli was present only in muscids (14.3%) and caliphorids (33.3%). Salmonella spp. was isolated from 9.5% of the muscids, 7.4% of the caliphorids, and 14.29% of the sarcophagids. Staphylococcus spp. was isolated from 28.5% of the muscids and 29.6% of the caliphorids. E. coli isolation was more frequent in flies captured on farms at which domestic chickens were housed near the milking parlors (p = 0.031) and which did not use sugarcane for animal feed (p = 0.042). Two out of 27 (7.4%) Staphylococcus spp. strains presented a phenotype of coagulase enzyme production. Ceftriaxone, ciprofloxacin, enrofloxacin, and gentamicin were the most effective antibiotics against E. coli and Salmonella spp. Tetracycline was the least effective antibiotic against the isolates. The Staphylococcus spp. strains isolated did not show the phenotype of oxacillin resistance. We conclude that regional flies are potential mechanical vectors of microbial agents able to cause enteritis in goats, mastitis in cows, and contamination of milk products for human consumption.