scholarly journals Proteomics characterization of Staphylococcus spp. from goat mastitis and phenogeno-typical assessment of resistance to beta-lactamics

2021 ◽  
Vol 41 ◽  
Author(s):  
Camila S. Pereira ◽  
Lídia M.M. Santos ◽  
Leandro S. Machado ◽  
Dayanne A. Melo ◽  
Shana M.O. Coelho ◽  
...  
Keyword(s):  
Clavulanic Acid ◽  
Meca Gene ◽  
Disk Diffusion ◽  
Penicillin G ◽  
Rrna Gene ◽  
Disk Diffusion Test ◽  
Maldi Tof ◽  
Two Samples ◽  
Amoxicillin Clavulanic Acid ◽  
Staphylococcus Spp

ABSTRACT: Mastitis occupies a prominent place among the diseases that affect dairy herds due to economic problems and public health. Staphylococcus spp. are infectious agents more involved in the etiology of caprine mastites, especially coagulase-negative Staphylococcus. Nineteen isolates of Staphylococcus spp. were obtained from subclinical caprine mastitis. All isolates were characterized by MALDI-TOF MS, being 47.36% (9/19) identified for S. epidermidis, 15.78% (3/19) for S. warneri, 10.52% (2/19) for S. aureus and S. caprae and 5.26% (1/19) for S. lugdunensis, S. simulans, and S. cohnii. All isolates characterized by MALDI-TOF were subjected a to polymerase chain reaction (PCR) for the 16S rRNA gene of Staphylococcus spp. to confirm the gender. After determining the species, tests for phenotypic detection of resistance to beta-lactams were carried out simple disk diffusion oxacillin, cefoxitin, penicillin G and amoxicillin + clavulanic acid, agar “screen” oxacillin and microdilution (MIC) cefoxitin. The disk diffusion test showed a strength of 58% (11/19) for penicillin G, 26.31% (5/19) for cefoxitin and 26.31% (5/19) for oxacillin. All strains were susceptible to amoxicillin + clavulanic acid and agar “screen” oxacillin. In the MIC, 63.15% (12/19) of the samples were cefoxitin resistant (MIC >4.0μg/ml). Then isolates were subjected to detection of the mecA resistance genes and regulators (mecl and mecRI), mecC and blaZ. Two samples of Staphylococcus epidermidis had the mecA gene. All isolates were negative for the mecA gene variant, mecl, mecRI, mecC and blaZ. These findings reinforce the importance of this group of microorganisms in the etiology of subclinical mastitis in goats and open perspectives for future research to investigate the epidemiology of the disease.

scholarly journals MULTIPLEX-PCR FOR DETECTION OF β-LACTAM RESISTANCE IN Staphylococcus spp.

2019 ◽  
Vol 6 (2) ◽  
pp. 262-275
Author(s):  
Giovana Hashimoto Nakadomari ◽  
Amanda Carmen Charalo ◽  
Ana Claudia Lemes Pavan ◽  
Vanessa Kelly Capoia Vignoto ◽  
Ricardo Antonio Pilegi Sfaciotte ◽  
...  
Keyword(s):  
Multiplex Pcr ◽  
Resistance Genes ◽  
Internal Control ◽  
Meca Gene ◽  
Simultaneous Detection ◽  
Rrna Gene ◽  
16S Gene ◽  
Internal Reaction ◽  
Two Samples ◽  
Staphylococcus Spp

A Staphylococcus Multiplex PCR system was developed for the simultaneous detection of the mecA, mecC, blaZ (resistance genes of b-lactam resistance) and PVL (pathogenicity factor gene), associated with an internal reaction control with the 16S rRNA gene. There were used primers described in the literature with and without modification and designed primers to standardize the hybridization and amplification temperature of distinct bands with 139 bp (mecC), 228 bp (16S), 313 bp (mecA), 408 bp (PVL) and 516 bp (blaZ) of molecular weight. The standardization was performed in ATCC strains and Staphylococcus schleiferiand tested in 56 strains of Staphylococcusspp. The 16S gene (internal control) was amplified in all samples, mecA gene was detected in two samples, mecA associated with mecC gene in one sample, mecA associated to the blaZ gene in 14 samples and the blaZ gene in 15 samples. No resistance genes were amplified in 24 samples. The PVL gene was not amplified in any of the samples tested.

scholarly journals Finding of coagulase negative staphylococci in the herd with an increased number of somatic cells in milk and their antimicrobial susceptibility

2013 ◽  
Vol 67 (3-4) ◽  
pp. 175-185
Author(s):  
Vera Katic ◽  
Natasa Rajic-Savic
Keyword(s):  
Clavulanic Acid ◽  
Somatic Cells ◽  
Inhibition Zone ◽  
Subclinical Mastitis ◽  
Blood Agar ◽  
Penicillin G ◽  
Coagulase Negative Staphylococci ◽  
Milk Samples ◽  
Causative Agents ◽  
Amoxicillin Clavulanic Acid

Coagulase-negative staphylococci (CNS) are generally considered to be opportunistic pathogens. Controlling CNS mastitis is difficult because the epidemiology is not clear, and the CNS group consists of about 40 different Staphylococcus species. Therefore, the aim of this study was to determine the prevalence of coagulasenegative staphylococci in milk of the cows with subclinical mastitis, as well as to determine different CNS species isolated from quarter milk samples for their susceptibility to antimicrobials used commonly for mastitis therapy. On the farm where there was found an increase of somatic cells in bulk milk, 112 dairy cows were examined by mastitis test. From 52 udder quarters where mastitis test showed an increase of somatic cells, milk samples were taken for bacteriological examination. For isolating the causes of mastitis there was used blood agar. Identification of the causative agents of mastitis was carried out on the basis of colony appearance on blood agar and their physiological characteristics. Coagulasepositive staphylococci sensitivity which cause mastitis was tested by Kirby Bauer method. For susceptibility testing there were used commercially produced discs containing: 10 IU penicillin, amoxicillin/clavulanic acid (20 +10 ?g), cloxacillin 25 ?g, 30 ?g amoxicillin, cephalexin 30 ?g, ceftiofur 30 ?g, 15 ?g lincomycin, gentamicin and tetracycline 30 ?g. The sensitivity of microorganisms was evaluated on the basis of inhibition zone diameter recommended by the manufacturer and was labeled as sensitive (S) moderately sensitive (I) or resistant (R). Coagulase-negative staphylococci were isolated from 61.53% of samples from cows with subclinical mastitis, making them the most common cause of subclinical mastitis. The highest resistance of coagulase-negative staphylococci was found to penicillin G (58.33% of isolates). Full sensitivity of coagulase-negative staphylococci was found to amoxicillin / clavulanic acid (100% of isolates), a good sensitivity to ceftiofur (83.33% of isolates), cefalexin (70.83% of isolates) and ceftriaxone (41.66% of isolates).

scholarly journals Short- and long-term effects of amoxicillin/clavulanic acid or doxycycline on the gastrointestinal microbiome of growing cats

2021 ◽  
Author(s):  
Evangelia Stavroulaki ◽  
Jan S. Suchodolski ◽  
Rachel Pilla ◽  
Geoffrey T. Fosgate ◽  
Chi-Hsuan Sung ◽  
...  
Keyword(s):  
Clavulanic Acid ◽  
Antibiotic Treatment ◽  
Control Group ◽  
Rrna Gene ◽  
Upper Respiratory Tract ◽  
Long Term Effects ◽  
Microbial Composition ◽  
Increased Risk ◽  
Group 15 ◽  
Amoxicillin Clavulanic Acid

Antibiotic treatment in early life influences gastrointestinal (GI) microbial composition and function. In humans, the resultant intestinal dysbiosis is associated with an increased risk for certain diseases later in life. The objective of this study was to determine the temporal effects of antibiotic treatment on the GI microbiome of young cats. Fecal samples were collected from cats randomly allocated to receive either amoxicillin/clavulanic acid (20 mg/kg q12h) for 20 days (AMC group; 15 cats) or doxycycline (10 mg/kg q24h) for 28 days (DOX group;15 cats) as part of the standard treatment of upper respiratory tract infection. In addition, feces were collected from healthy control cats (CON group;15 cats). All cats were approximately two months of age at enrolment. Samples were collected on days 0 (baseline), 20 or 28 (AMC and DOX, respectively; last day of treatment), 60, 120, and 300. DNA was extracted and sequencing of the 16S rRNA gene and qPCR assays were performed. Fecal microbial composition was different on the last day of treatment for AMC cats, and 1 month after the end of antibiotic treatment for DOX cats, compared to CON cats. Species richness was significantly greater in DOX cats compared to CON cats on the last day of treatment. Abundance of Enterobacteriales was increased, and that of Erysipelotrichi was decreased in cats of the AMC group on the last day of treatment compared to CON cats. The abundance of the phylum Proteobacteria was increased in cats of the DOX group on days 60 and 120 compared to cats of the CON group. Only minor differences in abundances between the treatment groups and the control group were present on day 300. Both antibiotics appear to delay the developmental progression of the microbiome, and this effect is more profound during treatment with amoxicillin/clavulanic acid and one month after treatment with doxycycline. Future studies are required to determine if these changes influence microbiome function and whether they have possible effects on disease susceptibility in cats.

scholarly journals Ceftaroline resistance in Staphylococcus pseudintermedius gene mecA carriers

2018 ◽  
Vol 38 (12) ◽  
pp. 2233-2236
Author(s):  
Carolina B. Scherer ◽  
Larissa S. Botoni ◽  
Antônio U. Carvalho ◽  
Kelly M. Keller ◽  
Adriane P. Costa-Val
Keyword(s):  
Methicillin Resistance ◽  
Meca Gene ◽  
Disk Diffusion ◽  
Diffusion Test ◽  
Staphylococcus Pseudintermedius ◽  
Ceftaroline Fosamil ◽  
Disk Diffusion Test ◽  
Methicillin Resistant ◽  
Resistant Strains ◽  
Current Report

ABSTRACT: Infections caused by methicillin-resistant Staphylococcus aureus (MRSA) being a constant concern, ceftaroline fosamil has been recently approved as a new cephalosporin, active against MRSA, for use in humans; only rare cases of resistance have been reported till date. There is no report of resistance to ceftaroline in Staphylococcus pseudintermedius, which is the main bacterium causing dermatitis and otitis in dogs. To evaluate staphylococcal resistance to ceftaroline, 35 isolates of methicillin-resistant S. pseudintermedius (MRSP), carrying the mecA gene, from 26 dogs with folliculitis and nine dogs with external otitis, underwent disk diffusion test with cefoxitin, oxacillin, and ceftaroline. Tests with cefoxitin and oxacillin showed > 90% sensitivity in methicillin resistance detection. In the disk diffusion test, 97.14% (34/35) were resistant to cefoxitin, 94.29% (33/35) to oxacillin, and 31.43% (11/35) to ceftaroline. Of the ceftaroline-resistant strains, 27.27% (3/11) were obtained from the ears of dogs while the rest (8/11) were from the skin. The current report is the first description of MRSP resistance to ceftaroline.

scholarly journals Antimicrobial Susceptibility of Clinically Relevant Gram-Positive Anaerobic Cocci Collected over a Three-Year Period in the Netherlands

2010 ◽  
Vol 55 (3) ◽  
pp. 1199-1203 ◽  
Author(s):  
A. C. M. Veloo ◽  
G. W. Welling ◽  
J. E. Degener
Keyword(s):  
The Netherlands ◽  
Clavulanic Acid ◽  
Antimicrobial Susceptibility ◽  
Penicillin G ◽  
Correct Identification ◽  
Gram Positive ◽  
Parvimonas Micra ◽  
Amoxicillin Clavulanic Acid

ABSTRACTThe susceptibility of 14 species of 115 Gram-positive anaerobic cocci (GPAC) was determined for 14 antibiotics. To assure correct identification, strains were genotypically identified by fluorescencein situhybridization and sequencing. Susceptibility differences (MIC50and MIC90) for penicillin G, clindamycin, tigecycline, levofloxacin, amoxicillin-clavulanic acid, cefoxitin, ertapenem, meropenem, metronidazole, and doxycycline were found for the three clinically most relevant GPAC species:Finegoldia magna,Parvimonas micra, andPeptoniphilus harei.

scholarly journals COMPARISON BETWEEN DOUBLE DISK SYNERGY TEST (DDST) AND INHIBITORY POTENTIATED DISK DIFFUSION TEST (IPDDT) FOR DETECTION OF ESBL ISOLATES.

2021 ◽  
pp. 130-132
Author(s):  
Sushil Kumar Sahu ◽  
Anshu Sharma ◽  
Neha Agrawal ◽  
Subhangi Sharma
Keyword(s):  
Clavulanic Acid ◽  
Pleural Fluid ◽  
Disk Diffusion ◽  
Clinical Samples ◽  
Esbl Production ◽  
Diffusion Test ◽  
Klebsiella Species ◽  
Disk Diffusion Test ◽  
E Coli ◽  
Synergy Test

Objective: To establish effectiveness between Double Disk Synergy Test (DDST) and Inhibitory Potentiated Disk Diffusion Test (IPDDT) for detection of ESBL isolates.Material and Method: Total two hundred screened isolation of E. coli and Klebsiella species obtained from 3146 various clinical samples such as pus, urine, blood, CSF, ear discharge, pleural fluid & sputum were included in the study. ESBL production where detected by Double Disk Synergy Test (DDST) and Inhibitory Potentiated Disk Diffusion Test (IPDDT) as per CLSI guidelines.Results: 200 isolates 117(58.5 %) E.coli and 83(41.5%) Klebsiella were identified.By DDST test 92.6% & 89.1%,66.7% & 71.7%,88.8% & 71.7% showed synergy between Ceftazidime, Cefotaxime, Ceftriaxone and Amoxicillin with Clavulanic acid by Klebsiella species and E.coli respectively.By IPDDT test 27.7% & 40.1%,28.9% & 43.4% and 36.1% & 44.4% showed synergy between Ca Vs Ca/C , Ce Vs Ce/C ,Ca Vs Ca/C + Ce Vs Ce/C by Klebsiella species and E.coli respectively.The ESBL isolate detected by DDST was 73(36.5%) and by IPDDT was 82 (41%). Conclusion: Additional 4.5% (9) ESBL isolates detected by IPDDT than DDST. ESBL detection have a significant impact on several important clinical outcomes. ESBLs outbreaks require accurate and precise detection.

scholarly journals β-Lactamase Production and Antimicrobial Susceptibility of Oral Heterogeneous Fusobacterium nucleatum Populations in Young Children

1999 ◽  
Vol 43 (5) ◽  
pp. 1270-1273 ◽  
Author(s):  
E. Könönen ◽  
A. Kanervo ◽  
K. Salminen ◽  
H. Jousimies-Somer
Keyword(s):  
Young Children ◽  
Clavulanic Acid ◽  
Antimicrobial Susceptibility ◽  
Fusobacterium Nucleatum ◽  
Penicillin G ◽  
Healthy Children ◽  
Good Activity ◽  
In Vitro Susceptibility ◽  
Amoxicillin Clavulanic Acid

ABSTRACT Oral Fusobacterium nucleatum populations from 20 young, healthy children were examined for β-lactamase production. Ten children (50%) harbored, altogether, 25 β-lactamase-positiveF. nucleatum isolates that were identified as F. nucleatum subsp. polymorphum, F. nucleatum subsp. nucleatum, and F. nucleatum subsp. vincentii (J. L. Dzink, M. T. Sheenan, and S. S. Socransky, Int. J. Syst. Bacteriol. 40:74–78, 1990). In vitro susceptibility of these β-lactamase-producing and 26 non-β-lactamase-producing F. nucleatum isolates was tested with penicillin G, amoxicillin-clavulanic acid, tetracycline hydrochloride, metronidazole, trovafloxacin, and azithromycin. Except for penicillin G, the antimicrobials exhibited good activity against all F. nucleatum isolates.

scholarly journals Phenotypic and Genotypic Characterization with MALDI-TOF-MS Based Identification of Staphylococcus spp. Isolated from Mobile Phones with their Antibiotic Susceptibility, Biofilm Formation, and Adhesion Properties

2020 ◽  
Vol 17 (11) ◽  
pp. 3761 ◽  
Author(s):  
Emira Noumi ◽  
Abderrahmen Merghni ◽  
Mousa Alreshidi ◽  
Rosa Del Campo ◽  
Mohd Adnan ◽  
...  
Keyword(s):  
Mobile Phones ◽  
Meca Gene ◽  
Hydrolytic Enzymes ◽  
Adhesion Properties ◽  
Maldi Tof Ms ◽  
Maldi Tof ◽  
Wide Range ◽  
The Matrix ◽  
Staphylococcus Spp ◽  
Tof Ms

Cell phones, smartphones, and tablets are extensively used in social and professional life, so they are frequently exposed to bacteria. The main goal of the present work was to isolate and characterize Staphylococci strains from students’ cell phone mobiles. Subsequently, 24 Staphylococci strains were tested against a wide range of antibiotics, for the distribution of some virulence-related genes and their ability to form biofilm. Staphylococcus spp. were cultured from all studied devices on chromogenic medium and identified using the matrix-assisted laser desorption/ionization (MALDI), time-of-flight (TOF) mass spectrometry (MS) technique (MALDI-TOF-MS). The results obtained showed that S. aureus was the dominant species (19 strains, 79.1%), followed by S. warneri (3 strains, 12.5%), and S. haemolyticus (2 strains, 8.3%). Isolated strains showed high percentages of hydrolytic enzymes production, resistance to many tested antibiotics, and 37.5% expressed the mecA gene. The tested strains were highly adhesive to polystyrene and glass and expressed implicated icaA (62.5%) and icaD (66.6%) genes. All Staphylococcus spp. strains tested were found to possess proteases and the α-hemolysin gene. Our results highlighted the importance of mobile phones as a great source of Staphylococcus spp., and these species were found to be resistant to many antibiotics with multiple antibiotic resistance (MAR) index ranging from (0.444) to (0.812). Most of the studied strains are able to form biofilm and expressed many virulence genes. Phylogenetic analysis based on the phenotypic and genetic characters highlighted the phenotypic and genetic heterogeneity of the S. aureus population studied. Further analyses are needed to elucidate the human health risks associated with the identified Staphylococci strains.

scholarly journals Susceptibility of Incisional Site Staphylococcus spp. to Three Common Perioperative Antimicrobials in Dogs Undergoing Clean Orthopaedic Procedures

VCOT Open ◽  
2020 ◽  
Vol 03 (02) ◽  
pp. e158-e163
Author(s):  
Po-Yen Chou ◽  
Duane Robinson ◽  
Amy S. Kapatkin ◽  
Steven E. Epstein ◽  
Michelle A. Giuffrida ◽  
...  
Keyword(s):  
Clavulanic Acid ◽  
Prospective Observational Study ◽  
Bacterial Isolates ◽  
Susceptibility Pattern ◽  
Coagulase Negative Staphylococci ◽  
Bacillus Spp ◽  
Amoxicillin Clavulanic Acid ◽  
Healthy Dogs ◽  
Staphylococcus Spp ◽  
Coagulase Positive Staphylococci

Abstract Objectives The aim of this study was to evaluate the susceptibility pattern of the incisional site Staphylococcus spp. to three commonly used perioperative antimicrobials in dogs. Materials and Methods A cohort of client-owned dogs was included in this prospective observational study. After induction of general anaesthesia and hair clipping, culture samples were collected from the incisional site of healthy dogs before orthopaedic surgery. The isolated colonies were identified. The susceptibility pattern of staphylococcal isolates was determined using Clinical and Laboratory Standard Institute standards. The staphylococcal susceptibilities to amoxicillin/clavulanic acid and cefpodoxime were compared with cefazolin using two-sided equality of proportions test. Results A total of 391 bacterial isolates were identified in 115 dogs. Bacillus spp. (n = 200) and Staphylococcus spp. (n = 95) were the most common bacteria isolated. Coagulase- positive and -negative staphylococci were isolated in 44 and 38% of dogs respectively. Cefazolin, amoxicillin/clavulanic acid and cefpodoxime staphylococcal susceptibilities were 68, 77 and 76% respectively. There were no significant differences in Staphylococcus spp. susceptibility between amoxicillin/clavulanic acid or cefpodoxime and cefazolin (p = 0.19, p = 0.25). Thirty-six per cent of coagulase-negative staphylococci and 6% of coagulase-positive staphylococci isolated were methicillin resistant. Conclusion Cefazolin, amoxicillin/clavulanic acid and cefpodoxime had similar susceptibilities for staphylococcal isolates. Routine use of amoxicillin/clavulanic acid or cefpodoxime in replace of cefazolin should be practiced with caution.

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