scholarly journals Characterization of Staphylococci and Streptococci Isolated from Milk of Bovides with Mastitis in Egypt

Pathogens ◽  
2020 ◽  
Vol 9 (5) ◽  
pp. 381
Author(s):  
Wedad Ahmed ◽  
Heinrich Neubauer ◽  
Herbert Tomaso ◽  
Fatma Ibrahim El Hofy ◽  
Stefan Monecke ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Microarray Analysis ◽  
Streptococcus Agalactiae ◽  
Nile Delta ◽  
Meca Gene ◽  
Resistance Rate ◽  
Coagulase Negative Staphylococci ◽  
Flight Mass Spectrometry ◽  
Streptococcus Gallolyticus ◽  
Broth Microdilution Method

The aim of this study was to characterize staphylococci and streptococci in milk from Egyptian bovides. In total, 50 milk samples were collected from localities in the Nile Delta region of Egypt. Isolates were cultivated, identified using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS), and antibiotic susceptibility testing was performed by the broth microdilution method. PCR amplifications were carried out, targeting resistance-associated genes. Thirty-eight Staphylococcus isolates and six Streptococcus isolates could be cultivated. Staphylococcus aureus isolates revealed a high resistance rate to penicillin, ampicillin, clindamycin, and erythromycin. The mecA gene defining methicillin-resistant Staphylococcus aureus, erm(C) and aac-aphD genes was found in 87.5% of each. Coagulase-negative staphylococci showed a high prevalence of mecA, blaZ and tetK genes. Other resistance-associated genes were found. All Streptococcus dysgalactiae isolates carried blaZ, erm(A), erm(B), erm(C) and lnuA genes, while Streptococcus suis harbored erm(C), aphA-3, tetL and tetM genes, additionally. In Streptococcus gallolyticus, most of these genes were found. The Streptococcus agalactiae isolate harbored blaZ, erm(B), erm(C), lnuA, tetK, tetL and tetM genes. Streptococcus agalactiae isolate was analyzed by DNA microarray analysis. It was determined as sequence type 14, belonging to clonal complex 19 and represented capsule type VI. Pilus and cell wall protein genes, pavA, cadD and emrB/qacA genes were identified by microarray analysis.

scholarly journals Characterization of Enterococci- and ESBL-Producing Escherichia coli Isolated from Milk of Bovides with Mastitis in Egypt

Pathogens ◽  
2021 ◽  
Vol 10 (2) ◽  
pp. 97
Author(s):  
Wedad Ahmed ◽  
Heinrich Neubauer ◽  
Herbert Tomaso ◽  
Fatma Ibrahim El Hofy ◽  
Stefan Monecke ◽  
...  
Keyword(s):  
Microarray Analysis ◽  
Dna Microarray ◽  
Nile Delta ◽  
Bovine Mastitis ◽  
Resistance Rate ◽  
E Coli ◽  
Dna Microarray Analysis ◽  
Vana Gene ◽  
Microdilution Method ◽  
Broth Microdilution Method

This study aimed to investigate the prevalence and antimicrobial resistance of enterococci- and ESBL-producing E. coli isolated from milk of bovine mastitis cases in Egypt. Fifty milk samples of dairy animals were collected from localities in the Nile Delta region of Egypt. Isolates were identified using MALDI-TOF MS, and antibiotic susceptibility testing was performed by the broth microdilution method. PCR amplifications were carried out, targeting resistance-associated genes. Seventeen Enterococcus isolates and eight coliform isolates could be cultivated. Vancomycin resistance rate was high in Ent. faecalis. The VITEK 2 system confirmed all E. coli isolates as ESBL-producing. All Ent. faecalis isolates harbored erm(B), tetL and aac-aphD genes. The vanA gene was detected in Ent. faecalis isolate, vanB was found in other Enterococcus, while one isolate of E. casseliflavus exhibited the vanA gene. E. coli isolates exhibited high prevalence of erm(B) and tetL. E. coli isolates were analyzed by DNA microarray analysis. Four isolates were determined by O-serotyping as O8 (n = 1), O86 (n = 2) and O157 (n = 1). H-serotyping resulted in H11, H12, H21 (two isolates each) and one was of H16 type. Different virulence-associated genes were detected in E. coli isolates including lpfA, astA, celB, cmahemL, intI1 and intI2, and the iroN gene was identified by DNA microarray analysis.

scholarly journals Whole Genome Sequencing and Antimicrobial Resistance of Staphylococcus aureus from Surgical Site Infections in Ghana

Pathogens ◽  
2021 ◽  
Vol 10 (2) ◽  
pp. 196
Author(s):  
Beverly Egyir ◽  
Jeannette Bentum ◽  
Naiki Attram ◽  
Anne Fox ◽  
Noah Obeng-Nkrumah ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Whole Genome Sequencing ◽  
Genome Sequencing ◽  
Meca Gene ◽  
Surgical Site Infections ◽  
Illumina Miseq ◽  
Multi Drug Resistance ◽  
Toxin Gene ◽  
Whole Genome ◽  
Flight Mass Spectrometry

Staphylococcus aureus (S. aureus) is a common cause of surgical site infections (SSIs) globally. Data on the occurrence of methicillin-susceptible S. aureus (MSSA) as well as methicillin-resistant S. aureus (MRSA) among patients with surgical site infections (SSIs) in sub-Saharan African are scarce. We characterized S. aureus from SSIs in Ghana using molecular methods and antimicrobial susceptibility testing (AST). Wound swabs or aspirate samples were collected from subjects with SSIs. S. aureus was identified by matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF-MS); AST was performed by Kirby-Bauer disk diffusion, and results were interpreted according to the Clinical and Laboratory Standards Institute (CLSI) guideline. Detection of spa, mecA, and pvl genes was performed by polymerase chain reaction (PCR). Whole-genome sequencing (WGS) was done using the Illumina MiSeq platform. Samples were collected from 112 subjects, with 13 S. aureus isolates recovered. Of these, 92% were sensitive to co-trimoxazole, 77% to clindamycin, and 54% to erythromycin. Multi-drug resistance was detected in 5 (38%) isolates. The four mecA gene-positive MRSA isolates detected belonged to ST152 (n = 3) and ST5 (n = 1). In total, 62% of the isolates were positive for the Panton-Valentine leukocidin (pvl) toxin gene. This study reports, for the first time, a pvl-positive ST152-t355 MRSA clone from SSIs in Ghana. The occurrence of multi-drug-resistant S. aureus epidemic clones suggests that continuous surveillance is required to monitor the spread and resistance trends of S. aureus in hospital settings in the country.

Bacteriocin ST91KM, produced by Streptococcus gallolyticus subsp. macedonicus ST91KM, is a narrow-spectrum peptide active against bacteria associated with mastitis in dairy cattle

2008 ◽  
Vol 54 (7) ◽  
pp. 525-531 ◽  
Author(s):  
Reneé Pieterse ◽  
Svetoslav D. Todorov ◽  
Leon M.T. Dicks
Keyword(s):  
Staphylococcus Aureus ◽  
Antimicrobial Activity ◽  
Streptococcus Agalactiae ◽  
Staphylococcus Epidermidis ◽  
Mode Of Action ◽  
Proteolytic Enzymes ◽  
Streptococcus Dysgalactiae ◽  
Streptococcus Gallolyticus ◽  
Streptococcus Uberis ◽  
Sds Page

Streptococcus gallolyticus subsp. macedonicus ST91KM produces a bacteriocin (macedocin ST91KM) active against Streptococcus agalactiae , Streptococcus dysgalactiae subsp. dysgalactiae , Streptococcus uberis , Staphylococcus aureus , and Staphylococcus epidermidis . Macedocin ST91KM is, according to tricine-SDS PAGE, between 2.0 and 2.5 kDa in size. Antimicrobial activity remained unchanged after 2 h of incubation at pH 2.0–10.0 and after 100 min at 100 °C. The peptide was inactivated after 20 min at 121 °C and when treated with proteolytic enzymes. Treatment with α-amylase had no effect on activity, suggesting that the mode of action does not depend on glycosylation. Amplification of the genome of strain ST91KM with primers designed from the macedocin precursor gene (mcdA) produced 2 fragments (approximately 375 and 220 bp) instead of one 150-bp fragment, as recorded for macedocin produced by Streptococcus gallolyticus subsp. macedonicus ACA-DC 198. Strain ACA-DC 198 was not available. However, DNA amplified from strain LMG 18488 (ACA-DC 206), genetically closely related to strain ACA-DC 198, revealed 99% homology to the mcdA of strain ACA-DC 198 (accession No. DQ835394). Macedocin ST91KM may thus be a second putative bacteriocin described for Streptococcus gallolyticus subsp. macedonicus.

scholarly journals Antimicrobial Resistance of Staphylococcus sp. Isolated from Cheeses

Animals ◽  
2021 ◽  
Vol 12 (1) ◽  
pp. 36
Author(s):  
Jana Výrostková ◽  
Ivana Regecová ◽  
František Zigo ◽  
Boris Semjon ◽  
Gabriela Gregová
Keyword(s):  
Polymerase Chain Reaction ◽  
Antimicrobial Resistance ◽  
Meca Gene ◽  
Common Species ◽  
Dilution Method ◽  
Agar Dilution Method ◽  
Chain Reaction ◽  
Coagulase Negative Staphylococci ◽  
Flight Mass Spectrometry ◽  
Polymerase Chain

S. aureus and some species of coagulase-negative staphylococci, including S. chromogenes and S. simulans, commonly cause intramammary infections. However, little attention was paid to the antimicrobial resistance of these species with respect to their occurrence in dairy products, for example, popular sheep and goat cheeses made from unpasteurized milk. The aim of this study was to investigate such sheep and goat cheeses for the occurrence and antimicrobial resistance of the relevant staphylococci species. The staphylococcal isolates were identified by polymerase chain reaction (130 isolates) and matrix assisted laser desorption/ionization time-of-flight mass spectrometry. The most common species of S. aureus (56 isolates) were identified, as well as S. chromogenes (16 isolates) and S. simulans (10 isolates). Antimicrobial resistance to penicillin, oxacilin, ceftaroline, teicoplanin, gentamicin, erythromycin, tetracycline and ofloxacin was subsequently determined in these species using the agar dilution method. The highest resistance was confirmed in all species, especially to penicillin (91%) and erythromycin (67%). The highest sensitivity was confirmed to ofloxacin (83%). Due to the high incidence of penicillin and oxacilin-resistant staphylococci, the mecA gene was detected by polymerase chain reaction, which was confirmed only in S. aureus isolates (19%). Our study shows that the tested strains (77%) were resistant to more than one antibiotic at a time.

Bacterial Aetiology of Bloodstream Infection and Antimicrobial Resistance Pattern in a Tertiary Hospital of Northern Bangladesh: Analysis of Current Situation

2020 ◽  
Vol 4 (02) ◽  
pp. 33-38
Author(s):  
M. Morsed Zaman Miah ◽  
Md. About Rafi ◽  
Md. Azizul Haque ◽  
Md. Kh. Faisal Alam
Keyword(s):  
Staphylococcus Aureus ◽  
Blood Stream Infection ◽  
Positive Culture ◽  
Blood Stream ◽  
Salmonella Typhi ◽  
Resistance Rate ◽  
Resistance Pattern ◽  
Coagulase Negative Staphylococci ◽  
Sensitivity Pattern ◽  
Bacterial Aetiology

Background: The objective of the present study was to identify the causative organisms of blood stream infection (BSI) and their resistance pattern to different antibiotics as well as prevalence of multi drug resistant (MDR) organisms in this region. This retrospective study included blood culture reports from 1899 suspected bacteraemia patients. Culture was done using BACT/Alert machine followed by culture on MacConkey (MC) agar, chocolate agar and blood agar plates. Isolated organisms were identified using standard laboratory procedures. Results: Total 383 bacterial isolates were yielded (rate of positive culture 20.2%). Staphylococcus aureus (41.8%) and Escherichia coli (41.8%) were most frequently isolated gram positive and gram-negative organisms respectively. Other commonly isolated organisms were Salmonella typhi (10.7%) and coagulase negative Staphylococci (CoNS) (3.9%). More than 90% isolated organisms were multidrug resistant. Salmonella typhi (95.1%) and Staphylococcus aureus (91.2%) showed most frequently isolated MDR strains. All the organisms showed high resistance rate against commonly used antibiotics like azithromycin, ciprofloxacin, and trimethoprim/sulfamethoxazole. Amoxycillin and clavulanic acid combination, cloxacillin and linezolid were sensitive against Staphylococcus aureus. Ceftriaxone as well as amikacin remained a sensitive drug to treat Salmonella typhi. Carbapenems and nitrofurantoin were mostly sensitive against all isolated organisms. Conclusion: Rational use of antibiotics based on regional epidemiology of causative organisms and sensitivity pattern can preserve the potentiality of available antibiotics and reduce the burden of MDR pathogens.

scholarly journals Occurrence of Methicillin-Resistant Coagulase-Negative Staphylococci (MRCoNS) and Methicillin-Resistant Staphylococcus aureus (MRSA) from Pigs and Farm Environment in Northwestern Italy

Antibiotics ◽  
2021 ◽  
Vol 10 (6) ◽  
pp. 676
Author(s):  
Miryam Bonvegna ◽  
Elena Grego ◽  
Bruno Sona ◽  
Maria Cristina Stella ◽  
Patrizia Nebbia ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Human Health ◽  
Environmental Samples ◽  
Methicillin Resistant Staphylococcus Aureus ◽  
Bacterial Species ◽  
Meca Gene ◽  
Nucleotide Sequencing ◽  
Coagulase Negative Staphylococci ◽  
Methicillin Resistant ◽  
Nasal Swabs

Swine farming as a source of methicillin-resistant Staphylococcus aureus (MRSA) has been well documented. Methicillin-resistant coagulase-negative staphylococci (MRCoNS) have been less studied, but their importance as pathogens is increasing. MRCoNS are indeed considered relevant nosocomial pathogens; identifying putative sources of MRCoNS is thus gaining importance to prevent human health hazards. In the present study, we investigated MRSA and MRCoNS in animals and environment in five pigsties in a high farm-density area of northwestern Italy. Farms were three intensive, one intensive with antibiotic-free finishing, and one organic. We tested nasal swabs from 195 animals and 26 environmental samples from three production phases: post-weaning, finishing and female breeders. Phenotypic tests, including MALDI-TOF MS, were used for the identification of Staphylococcus species; PCR and nucleotide sequencing confirmed resistance and bacterial species. MRCoNS were recovered in 64.5% of nasal swabs, in all farms and animal categories, while MRSA was detected only in one post-weaning sample in one farm. The lowest prevalence of MRCoNS was detected in pigs from the organic farm and in the finishing of the antibiotic-free farm. MRCoNS were mainly Staphylococcus sciuri, but we also recovered S. pasteuri, S. haemolyticus, S. cohnii, S. equorum and S. xylosus. Fifteen environmental samples were positive for MRCoNS, which were mainly S. sciuri; no MRSA was found in the farms’ environment. The analyses of the mecA gene and the PBP2-a protein highlighted the same mecA fragment in strains of S. aureus, S. sciuri and S. haemolyticus. Our results show the emergence of MRCoNS carrying the mecA gene in swine farms. Moreover, they suggest that this gene might be horizontally transferred from MRCoNS to bacterial species more relevant for human health, such as S. aureus.

scholarly journals Reliability of mecA in Predicting Phenotypic Susceptibilities of Coagulase-Negative Staphylococci and Staphylococcus aureus

2020 ◽  
Vol 7 (12) ◽  
Author(s):  
Manon C Williams ◽  
Samuel R Dominguez ◽  
Andrea Prinzi ◽  
Kayla Lee ◽  
Sarah K Parker
Keyword(s):  
Staphylococcus Aureus ◽  
Meca Gene ◽  
Blood Cultures ◽  
Coagulase Negative Staphylococci ◽  
High Concordance ◽  
And Staphylococcus Aureus

Abstract The mecA gene is commonly used to identify resistance in Staphylococcus aureus, but historically is not used for coagulase-negative staphylococci (CoNS). Analysis of 412 staphylococcal blood cultures (2014–2018) revealed that the absence of mecA had high concordance (100%) with oxacillin susceptibility for S. aureus and CoNS alike.

scholarly journals Mannitol Fermenting Methicillin-Resistant Coagulase Negative Staphylococci Isolated From Diabetic Foot Infections

2020 ◽  
Vol 11 (4) ◽  
pp. 6095-6101
Author(s):  
Samira Fattah Hamid ◽  
Aza Bahadeen Taha
Keyword(s):  
Staphylococcus Aureus ◽  
Diabetic Foot ◽  
Methicillin Resistance ◽  
Meca Gene ◽  
Rrna Gene ◽  
Human Pathogens ◽  
Coagulase Negative Staphylococci ◽  
Mannitol Salt Agar ◽  
Methicillin Resistant ◽  
Diabetic Foot Infections

Detection of mannitol fermenting coagulase-negative staphylococci is frequently unnoticed when Staphylococcus aureus is screening in the laboratory. On the other hand, the emergence of coagulase-negative staphylococci as critical human pathogens need dependable methods for the identification of clinically significant coagulase-negative staphylococci to understand the epidemiology of infections caused by these bacteria. The study aimed to identify mannitol fermenting coagulase-negative staphylococci that assumed to be Staphylococcus aureus as they formed yellow colonies on Mannitol Salt agar plates. Samples were taken from eighty-four patients with diabetic foot infections. The specimen was cultured on Blood agar and Mannitol Salt agar. Mannitol fermenting coagulase-negative staphylococci isolates diagnosed through Vitek2 system then confirmed by detecting 16S rRNA gene and absence of the nuc gene. Antibiotic sensitivity and methicillin resistance were detected by Vitek2 system, then methicillin resistance was confirmed by Oxacillin Salt Agar Screen test and detection of the mecA gene. Out of 81 Staphylococcus isolated from foot and nose of diabetic foot patients, twenty isolates were mannitol fermenting coagulase-negative staphylococci, they related to following species; Staphylococcus haemolyticus, staphylococcus lentus, Staphylococcus xylosus, Staphylococcus lugdunensis, Staphylococcus hominis, Staphylococcus galinarum and Staphylococcus saprophyticus). The majority of them (85%) were phenotypically methicillin-resistant and genotypically harbouring mecA gene. 80% were resistant to Erythromycin, 70% to Clindamycin, 35% to Trimethoprim-Sulphamethoxazole, 30% to Gentamicin and Rifampicin, 15% to Levofloxacin and Teicoplanin. 30% expressed inducible clindamycin resistance.

scholarly journals Molecular detection and typing of methicillin-resistant Staphylococcus aureus and methicillin-resistant coagulase-negative staphylococci isolated from cattle, animal handlers, and their environment from Karnataka, Southern Province of India

2019 ◽  
Vol 12 (11) ◽  
pp. 1760-1768 ◽  
Author(s):  
Nimita Venugopal ◽  
Susweta Mitra ◽  
Rituparna Tewari ◽  
Feroze Ganaie ◽  
Rajeswari Shome ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Methicillin Resistant Staphylococcus Aureus ◽  
Methicillin Resistance ◽  
Animal Health ◽  
Sccmec Type ◽  
Meca Gene ◽  
Molecular Characteristics ◽  
Rrna Gene ◽  
Coagulase Negative Staphylococci ◽  
Methicillin Resistant

Background and Aim: Methicillin-resistant staphylococci are among the emerging pathogens which have become a threat to both human and animal health. The present investigation intended to examine the occurrence and the molecular characteristics of methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-resistant coagulase-negative staphylococci (MRCoNS) recovered from cattle, its handlers, and their environment. Materials and Methods: A total of 666 specimens were subjected to culture method and genus-specific polymerase chain reaction (PCR) for the identification of Staphylococcus. Methicillin resistance was substantiated by PCR identification of mecA and mecC resistance determinants. Species-specific identification of mecA positive isolates was conducted by multiplex PCR. The unidentified species were deciphered by 16S rRNA gene sequencing approach. The mecA positive isolates were further characterized by staphylococcal cassette chromosome mec (SCCmec) typing and multilocus sequence typing (MLST). Results: Duplex PCR identified 728 Staphylococcus isolates, of which 66 (9%) were positive for mecA gene. MRSA constituted 24% of the total mecA positive isolates. Among MRCoNS, Staphylococcus epidermidis (42%), and Staphylococcus haemolyticus (11%) were the most common species identified. Overall, 47% of the mecA positive isolates belonged to SCCmec type V. MLST analysis showed eight different sequence types (STs) among MRSA isolates of which five were novel STs. Among methicillin-resistant S. epidermidis, 19 different STs were found, of which nine novel STs were detected. Conclusion: The increase in the prevalence of mecA positive staphylococci, especially MRCoNS in cattle is a great concern in view of their transmission potential. Hence, continuous monitoring and molecular characterization of methicillin-resistant staphylococci should be elucidated in human and animal sectors so as to prevent the spread of these resistant pathogens.

scholarly journals Detection of Staphylococcus aureus, Streptococcus agalactiae and Escherichia coli in Brazilian mastitic milk goats by multiplex-PCR

2018 ◽  
Vol 38 (7) ◽  
pp. 1358-1364 ◽  
Author(s):  
Gustavo P. Machado ◽  
Rodrigo C. Silva ◽  
Felipe F. Guimarães ◽  
Anelise Salina ◽  
Hélio Langoni
Keyword(s):  
Escherichia Coli ◽  
Staphylococcus Aureus ◽  
Multiplex Pcr ◽  
Streptococcus Agalactiae ◽  
High Performance ◽  
Dairy Goat ◽  
Coagulase Negative Staphylococci ◽  
E Coli ◽  
Milk Samples ◽  
Microbiological Culture

ABSTRACT: This study evalueted the prevalence of Staphylococcus aureus, Streptococcus agalactiae and Escherichia coli in milk samples from 257 goats (513 half-udders) and ten bulk tanks, from ten dairy goat farms of São Paulo State, Brazil, by multiplex-PCR. The samples were screened by microbiological culture (gold-standard), and tested by different multiplex-PCR protocols for the detection of each bacterium. A total of 178 half-udders resulted positive by microbiological culture, with coagulase-negative staphylococci (70%), S. aureus (13.5%), S. intermedius (7.9%), and Enterobacteriaceae (4%) the prevalent pathogens. In other way, multiplex-PCR detected 173 pathogens in 151/523 (28.9%; CI95% 25.2-32.9%) milk samples 144/513 (28.1%) half-udders and 7/10 (70%) bulk tanks, with E. coli (86/162, 51.9%) and S. aureus (50/162, 30.9%) the prevalent ones in half-udders, and S. aureus (6/10, 60%) and E. coli (4/5, 36.4%) in bulk tanks. Multiplex-PCR showed a high performance for the detection of three bacteria at a time in mastitic goat milk direct from half-udders or bulk tanks. Thus, this multiplex-PCR protocol proved to be an adequate tool for the identification of the most common mastitis pathogens, independent of their phenotypic characteristics in the diagnosis of clinical mastitis in goats, allowing a continuous and better vigilance and monitoring the herd, being included in quality programs.

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