CoNS Isolates Research Articles

Antiseptics and mupirocin resistance in clinical, environmental, and colonizing coagulase negative Staphylococcus isolates

BackgroundCoagulase-Negative Staphylococci (CoNS) are opportunistic and nosocomial pathogens. The excessive use of antimicrobial agents, including antiseptics, represents one of the world’s major public health problems. This study aimed to test the susceptibility of CoNS to antiseptics.MethodsOut of 250 specimens collected from different sections of the hospital, 55 samples were identified as CoNS, categorized into three groups based on their sources: environmental samples (n = 32), healthcare worker carriers samples (n = 14), and clinical infection samples (n = 9). Isolates were examined for susceptibility to antibiotics and antiseptics, such as benzalkonium chloride (BC), cetyltrimethylammonium bromide (CTAB), and chlorhexidine digluconate (CHDG). Mupirocin and antiseptic resistance genes, as well as the mecA gene, were detected using polymerase chain reaction. CoNS isolates with notable resistance to antiseptics and antibiotics were identified using the API-Staph system.ResultsA high frequency of multidrug resistance among CoNS clinical infection isolates was observed. Approximately half of the CoNS isolates from healthcare workers were susceptible to CHDG, but 93% were resistant to BC and CTAB. The frequency of antiseptics and antibiotics resistance genes in CoNS isolates was as follows: qacA/B (51/55; 92.7%), smr (22/55; 40.0%), qacG (1/55; 1.8%), qacH (6/55; 10.9%), qacJ (4/55; 7.3%), mecA (35/55; 63.6%), mupB (10/55; 18.2%), and mupA (7/55; 12.7%). A significant difference in the prevalence of smr gene and qacJ genes between CoNS isolates from healthcare workers and other isolates was reported (P value = 0.032 and ˂0.001, respectively). Four different CoNS species; S. epidermidis, S. chromogene, S. haemolyticus, and S. hominis, were identified by API.ConclusionsCoNS isolates colonizing healthcare workers showed a high prevalence of antiseptic resistance genes, while clinical infection samples were more resistant to antibiotics. CHDG demonstrated greater efficacy than BC and CTAB in our hospital.

Study of coagulase-negative staphylococci in hospital-acquired neonatal sepsis

Background Coagulase-negative staphylococci (CoNS) has become increasingly prevalent as the leading cause of late-onset bacteremia in neonatal ICUs. Diagnosing CoNS septicemia poses challenges because this bacterium commonly resides on the skin, leading to potential contamination of blood culture samples. Therefore, this study aimed to identify the bacterial causes of sepsis in neonates who exhibit clinical signs of the condition, also to assess the methicillin susceptibility of CoNS through both phenotypic and molecular methodologies. Materials and methods This study was carried out at Mansoura University Children Hospital. Blood samples were obtained and directly cultured using the BACT/ALERT system. All isolates were identified using BD Phoenix system. CoNS isolates were subjected to a disk diffusion susceptibility test with cefoxitin 30 µg, serving as an initial screening test to identify methicillin resistance. Automated antibiotic susceptibility was done using BD Phoenix system followed by PCR testing to detect the presence of the mecA gene in resistant CoNS isolates. Results Among all the isolated micro-organisms, Klebsiella pneumoniae and CoNS were found to be significantly higher in early-onset sepsis and late-onset sepsis, respectively. Staphylococcus epidermidis constituted 30% of the isolates, with Staphylococcus haemolyticus and Staphylococcus hominis each accounting for 20%, while other Staphylococcus species made up the remaining 20%. Among the identified CoNS, 47.4% tested positive for the mecA gene. The presence of the mecA gene was significantly associated with a higher incidence of late-onset sepsis (P=0.033). Conclusion CoNS are the predominant culprits behind late-onset sepsis in hospitalized newborns. Notably, the presence of the mecA gene was significantly linked to a higher occurrence of late-onset sepsis. Our study has uncovered a concerning surge in antibiotic resistance genes within our community. This underscores the urgency of implementing an active antibiotic surveillance program and exploring alternative therapeutic strategies to effectively combat neonatal sepsis.

In-vitro assessment of staphylococci biofilms formed under biologically-relevant conditions and correlation to the biofilm genotype

Staphylococci have been implicated in chronic device-related infections due to their ability to form resistant biofilms on implanted medical devices. For a long time, two different mechanisms of biofilm formation were known in Staphylococcus spp., the ica-dependent biofilms in MSSA and CoNS and the ica-independent biofilms in MRSA. Recently, a new fibrin-based biofilm phenotype was identified when S. aureus isolates were allowed to construct biofilms in biologically-relevant conditions using plasma-coated surfaces and RPMI-1640 for biofilm development (RPMI-1640/Pl). In this study, 140 staphylococci clinical isolates (91 MRSA, 27 MSSA and 22 CoNS) were tested for biofilm formation, biofilm formers were selected and used to scrutinize the ability of RPMI-1640/Pl to support staphylococci biofilm formation. Results showed that, in RPMI-1640/Pl, the biofilm formation abilities of MRSA and MSSA isolates were non-significantly different compared to those formed in TSB and BHI, (Kruskal Wallis test, P = 0.3275 and 0.466 for MRSA and MSSA isolates, respectively). However, a significantly different biofilm formation ability was observed regarding the tested CoNS isolates (ANOVA test, P = 0.0006). Furthermore, biofilm formation in RPMI-1640/Pl under different incubation conditions was tested, and among the tested conditions, 48h of static incubation showed significantly elevated biofilm for both MRSA and MSSA. Finally, PCR was used to detect genes implicated in biofilm formation, and the genotypes were correlated to the biofilm formation ability in different tested conditions. In contrast to ordinary media, biofilm formation by staphylococci in RPMI-1640/Pl was positively correlated to coa, fnbA, fnbB and clfB.

Antimicrobial susceptibility and biofilm forming ability of staphylococci from subclinical buffalo mastitis.

The starting objective of this research communication was to determine the prevalence of subclinical mastitis in buffalo in Turkey. We also seeked to isolate and identify staphylococci, determine their antimicrobial susceptibilities and biofilm-forming abilities as well as investigating the presence of biofilm-related genes and microbial surface components recognizing adhesive matrix molecules. A total of 107 (66.9%) staphylococci (28 S. aureus and 79 coagulase-negative staphylococci, CoNS) were isolated from 160 mastitic milk samples collected from 200 lactating water buffalos. The staphylococci were especially resistant to beta-lactams except for cefoxitin but were less resistant to the other antimicrobials that were tested. Based on the Congo red agar method, 92.9% of the S. aureus and 70.9% of the CoNS isolates were positive for biofilm-forming ability, while all S. aureus and 97.5% of CoNS isolates were positive by a microtiter plate analysis. The presence of icaA and icaD genes was not always correlated with biofilm synthesis, and even in the absence of these genes, the isolates were able to synthesize biofilm.

In Vivo Effectiveness of Several Antimicrobial Locks To Eradicate Intravascular Catheter Coagulase-Negative Staphylococci Biofilms.

Tunneled central venous catheter (TCVC) related infection remains a challenge in the care of hemodialysis patients. We aimed to determine the best antimicrobial lock therapy (ALT) to eradicate coagulase-negative staphylococci (CoNS) biofilms. We studied the colonization status of the catheter every 30 days by quantitative blood cultures (QBC) drawn through all catheter lumens. Those patients with a significant culture (i.e.,100 to 1,000 CFU/mL) of a CoNS were classified as patients with a high risk of developing catheter-related bloodstream infections (CRBSI). They were assigned to receive daptomycin, vancomycin, teicoplanin lock solution, or the standard of care (SoC) (i.e., heparin lock). The primary endpoint was to compare eradication ability (i.e., negative QBC for 30 days after ending ALT) rates between different locks and the SoC. A second objective was to analyze the correlation between ALT exposure and isolation of CoNS with antimicrobial resistance. Daptomycin lock was associated with a significant higher eradication success than with the SoC: 85% versus 30% (relative risk [RR] = 14, 95% confidence interval [CI] = 2.4 - 82.7); followed by teicoplanin locks with a 83.3% success (RR = 11.7; 95% CI = 2 - 70.2). We observed CoNs isolates with a higher teicoplanin MIC in patients with repeated teicoplanin locks exposure (coefficient = 0.3; 95% CI = 0.11 - 0.47). However, teicoplanin MICs decreased in patients treated with vancomycin locks (coefficient = -0.56; 95% CI = -0.85 - -0.02). Methicillin-resistance decreased with accumulative ALT (RR = 0.82; 95% CI = 0.69 - 0.98). In this study, daptomycin locks achieve the highest eradication rate of CoNS from hemodialysis catheters in vivo.

646. Activity of Oritavancin and Comparator Agents Against Coagulase-negative Staphylococci Causing Bloodstream Infections in US Medical Centers (2017-2019)

Abstract Background Coagulase-negative staphylococci (CoNS) is a common organism group implicated in catheter-related bloodstream infection (BSI) and infective endocarditis. Prompt appropriate antimicrobial therapy is crucial for suspected or confirmed invasive infections. The in vitro activity of oritavancin (ORI) and comparators was evaluated against CoNS causing BSI in US medical centers. Methods 587 CoNS isolates (1/patient) were consecutively collected in 30 US centers in 2017-2019. Bacterial identification was performed by MALDI-TOF, and susceptibility testing using CLSI broth microdilution methodology in a central laboratory. CLSI breakpoints were applied for comparators and the ORI susceptible (S) breakpoint for S. aureus (≤0.12 mg/L) was used for in vitro comparison only. Results The most common species were S. epidermidis (Sepi; 62.4%; 366), followed by S. hominis (Shom; 13.1%; 77), S. capitis (Scap; 7.3%; 43), S. lugdunensis (Slug; 5.1%; 30), and S. haemolyticus (Shae; 4.3%; 25). 12 other species represented < 10 (1.5%) isolates each. Overall, 59.1% of isolates were methicillin-resistant (MR), with the highest rate in Sepi (73.2%), followed by Shae (68.0%), Shom (46.8%), and Scap (30.2%). No MR isolates were detected in Slug. ORI (MIC50/90, 0.06/0.12 mg/L) inhibited 96.1% of CoNS at ≤0.12 mg/L. Linezolid (LZD; MIC50/90, 1/1 mg/L; 96.4%S), daptomycin (DAP; MIC50/90, 0.25/0.5 mg/L; 100%S), and vancomycin (VAN; MIC50/90, 1/2 mg/L; 100%S) were also active against CoNS. ORI displayed similar MIC50 (0.03-0.06 mg/L) and MIC90 (0.12-0.25 mg/L) values against Sepi, Shom, Scap, and Shae, and inhibited 96.0%, 96.1%, 97.7%, and 84.0% of these isolates at ≤0.12 mg/L, respectively. All Slug isolates were inhibited by ORI at ≤0.015 mg/L. ORI inhibited 94.8% of all MRCoNS at ≤0.12 mg/L, and 95.5%, 94.4%, 92.3%, and 82.4% of MR Sepi, Shom, Scap, and Shae species, respectively. VAN, DAP, and LZD inhibited 100.0%, 100.0%, and 93.9% of MRCoNS isolates at their susceptible breakpoints, respectively. Conclusion ORI was highly active and inhibited ≥96% of all CoNS and individual species ( >10 isolates) at ≤0.12 mg/L, regardless of methicillin profile, except for Shae. VAN, DAP, and LZD were also active against CoNS causing BSI in US medical centres. Disclosures Cecilia G. Carvalhaes, MD, PhD, AbbVie: Grant/Research Support|Cidara: Grant/Research Support|Melinta: Grant/Research Support|Pfizer: Grant/Research Support Helio S. Sader, MD, PhD, AbbVie: Grant/Research Support|Cidara: Grant/Research Support|Melinta: Grant/Research Support|Nabriva Therapeutics: Grant/Research Support|Pfizer: Grant/Research Support Jennifer M. Streit, BS, MT(ASCP), Cidara: Grant/Research Support|GSK: Grant/Research Support|Melinta: Grant/Research Support|Shionogi: Grant/Research Support Rodrigo E. Mendes, PhD, AbbVie: Grant/Research Support|Cidara: Grant/Research Support|GSK: Grant/Research Support|Melinta: Grant/Research Support|Nabriva Therapeutics: Grant/Research Support|Office for Assistant Secretary of Defense for Health Affairs: Grant/Research Support|Pfizer: Grant/Research Support|Shionogi: Grant/Research Support|Spero Therapeutics: Grant/Research Support.

Emerging vancomycin-non susceptible coagulase negative Staphylococci associated with skin and soft tissue infections

BackgroundsObservable emergence of Vancomycin-Non susceptible Coagulase-negative Staphylococci (VNS-CoNS) associated with skin and soft tissue infections spreading among the urban and rural populace is gradually intensifying severe complications. The isolated VNS-CoNS were evaluated with Matrix-assisted Laser Desorption/ionization Time of Flight Mass Spectrometry (MALDI ToF MS) for species characterization and pan-antimicrobial resistance pattern.MethodsOut of 256 clinical samples collected including pus, abscess, ear swabs, eye swabs, and aspirates, 91 CoNS isolates were biotyped and further characterized with MALDI-TOF MS. Staphylococci marker genes, Vancomycin susceptibility, and biofilm assays were performed.ResultsOf 91 CoNS isolates, S.cohnii (2.3%), S.condimentii (3.4%), S. saprophyticus (6.7%), and S.scuri (21.1%) were characterized with MALDI-TOF with significant detection rate (99.4%; CI 95, 0.775–0.997, positive predictive values, 90.2%) compared to lower biotyping detection rate (p = 0.001). Hemolytic VNS-CoNS lacked nuc, pvl and spa genes from wound, ear, and aspirates of more 0.83 MARI clustered into a separate phylo-diverse group and were widely distributed in urban and peri-urban locations. MALDI TOF–MS yielded a high discriminatory potential of AUC-ROC score of 0.963 with true-positivity prediction. VNS-CoNS of MIC ≥ 16 µg/mL were observed among all the ages with significant resistance at 25th and 75th quartiles. More than 10.5% of CoNS expressed multi-antibiotic resistance with more than 8 µg/mL vancomycin cut-off values (p < 0.05).ConclusionAntibiotic resistant CoNS should be considered significant pathogens rather than contaminant. Biofilm producing VNS-S. sciuri and S. condimentii are potential strains with high pathological tropism for skin, soft tissues and wound infections, and these strains require urgent surveillance in peri-urban and rural communities.

Inducible clindamycin resistance in coagulase negative staphylococci

Induced clindamycin resistance poses a major problem for the use of this drug to treat staphylococcal infections. An infectious etiology is often implicated in the presence of CoNS in clinical specimens. A total of 183 isolates of Staphylococci spp. recovered from different clinical specimens were studied. A coagulase test was done to select coagulase negative Staphylococci, and they were confirmed by conventional microbiological techniques. Using the double disc approximation test, induced clindamycin resistance was detected (D-test). The D test was used to detect the inducible clindamycin resistance phenotype in erythromycin resistant and clindamycin sensitive isolates. A micro titer plate test was used to determine biofilm development in these isolates. One hundred and twenty eight isolates from various clinical specimens were identified as proven CoNS, which were distributed across six different species’. haemolyticus, the most common species of CoNS isolated, was isolated from all types of specimens. The present study revealed the strong association of biofilm with inducible clindamycin resistance among the CoNs isolates.

Bacteriocin-Like Inhibitory Substances in Staphylococci of Different Origins and Species With Activity Against Relevant Pathogens.

Bacteriocins are antimicrobial peptides with relevance in the modulation of human and animal microbiota that have gained interest in biomedical and biotechnological applications. In this study, the production of bacteriocin-like inhibitory substances (BLIS) was tested among a collection of 890 staphylococci of different origins (humans, animals, food, and the environment) and species, both coagulase-positive (CoPS, 238 isolates of 3 species) and coagulase-negative staphylococci (CoNS, 652 isolates of 26 species). Of the 890 staphylococci, 60 (6.7%) showed antimicrobial activity by the spot-on-lawn method against at least one of the 25 indicator bacteria tested. BLIS-producer (BLIS+) isolates were detected in 8.8% of CoPS and 6.0% of CoNS. The staphylococcal species with the highest percentages of BLIS+ isolates were S. chromogenes (38.5%), S. pseudintermedius (26.7%), and S. warneri (23.1%). The production of BLIS was more frequently detected among isolates of pets, wild animals, and food. Moreover, 13 BLIS+ isolates showed wide antimicrobial activiy spectrum, and 7 of these isolates (of species S. aureus, S. pseudintermedius, S. sciuri, and S. hominis) demonstrated antimicrobial activity against more than 70% of the indicator bacteria tested. The genetic characterization (by PCR and sequencing) of the 60 BLIS+ isolates revealed the detection of (a) 11 CoNS and CoPS isolates carrying putative lantibiotic-like genes; (b) 3 S. pseudintermedius isolates harboring the genes of BacSp222 bacteriocin; and (c) 2 S. chromogenes isolates that presented the gene of a putative cyclic bacteriocin (uberolysin-like), being the first report in this CoNS species. Antimicrobial susceptibility testing was performed in BLIS+ isolates and one-third of the CoNS isolates showed susceptibility to all antibiotics tested, which also lacked the virulence genes studied. These BLIS+ CoNS are good candidates for further characterization studies.

Characterization of biofilm producing methicillin resistant coagulase negative Staphylococci from India.

Methicillin-resistant coagulase-negative staphylococci (MR-CoNS) cause infectious diseases due to their potential to form biofilm and further colonization in hospital materials. This study evaluated the antibiotic susceptible phenotypes, biofilm-producing ability, and biofilm-associated genes (mecA, icaAD, bap, cna, and fnbA). Biofilm formation was detected through Congo red agar (CRA) method and MTP method. The presence of biofilm and associated genes in MR-CoNS were detected by PCR. A total of 310 (55.95%) isolates produced the biofilm. Among these isolates, Staphylococcus haemolyticus (34.83%), Staphylococcus epidermis (31.93%), Staphylococcus capitis (16.77%), Staphylococcus cohnii (10.96%), and Staphylococcus hominis (5.48%) were identified. The antimicrobial susceptibility pattern of CoNS isolates indicated resistance to cefoxitin (100%), erythromycin (94.8%), ciprofloxacin (66.7%), sulfamethoxazole/trimethoprim (66.7%), gentamicin (66.12%), and clindamycin (62.9%). Resistance rate to mupirocin was 48.5% in S. epidermidis and 38.9% in S. haemolyticus isolates. All isolates were sensitive to vancomycin and linezolid. The prevalence rates of icaAD, bap, fnbA, and cna were 18.06%, 12.5%, 47.4%, and 27.4%, respectively. icaAD and bap genes were detected in 18.06% and 12.5% of MR-CoNS isolates. fnbA and cna genes were detected in 47.41% and 27.41% of MRCoNS isolates. icaAD positive strains exhibited a significant increase in the biofilm formation compared with those that lacked icaAD (0.86 (0.42, 1.39) versus 0.36 (0.14, 0.75), respectively; P < 0.001). In conclusion, the majority of MR-CoNS isolates were biofilm producers, and S. capitis, which possessed icaAD genes, ranked as the great biofilm producer than other Staphylococcus. The study's findings are important to form a strategy to control biofilm formation as an alternative strategy to counter the spread of MR-CoNS in healthcare settings.

Reservoir of Antibiotic Residues and Resistant Coagulase Negative Staphylococci in a Healthy Population in the Greater Accra Region, Ghana.

Antimicrobial resistance threatens infectious disease management outcomes, especially in developing countries. In this study, the occurrence of resistant coagulase-negative staphylococci (rCoNS) and antibiotic residues in urine samples of 401 healthy individuals from Korle-Gonno (KG) and Dodowa (DDW) in Ghana was investigated. MALDI-ToF/MS with gram-staining techniques detected and identified the CoNS. SPE-LC-MS/MS detected and quantified nine commonly used antibiotics in the samples. The results showed 63 CoNS isolates detected in 47 (12%) samples, with S. haemolyticus (78%) and S. epidermidis (8%) being predominant. Most of the isolates (95%) were resistant to at least one antibiotic, with the highest resistance observed against sulphamethoxazole (87%). Resistance profiles in samples from DDW and KG were largely comparable, but with some differences. For instance, DDW isolates were more resistant to gentamicin (p = 0.0244), trimethoprim (p = 0.0045), and cefoxitin (p = 0.0078), whereas KG isolates were more resistant to erythromycin (p = 0.0356). Although the volunteers had not knowingly consumed antibiotics two weeks before sampling, antibiotic residues, ranging between 1.44–17000 ng mL−1 were identified in 22% of urine samples. Samples with antibiotic residues were likely to also contain rCoNS (89%). The most frequent antibiotics detected were tetracycline (63%) and ciprofloxacin (54%). Healthy individuals could thus be reservoirs of antibiotic residues and rCoNS at the community level.

Cefazolin susceptibility of coagulase-negative staphylococci (CoNS) causing late-onset neonatal bacteraemia.

CoNS bacteraemia causes significant neonatal morbidity. Previous work has suggested that β-lactam antibiotics vary in their binding affinity to PBP2a (produced by the mecA gene) present in most CoNS. We evaluated cefazolin MICs for CoNS isolated in an Australian neonatal ICU (NICU) and correlated them with isolate genotype and phenotype. Significant blood isolates from 2009 to 2017 were speciated and underwent broth microdilution testing for cefazolin, cefoxitin, oxacillin and flucloxacillin. Correlation with mecA presence and PBP2a expression was evaluated. A selection of Staphylococcus capitis isolates underwent WGS. The CoNS (n = 99) isolates were confirmed as S. capitis (n = 57), Staphylococcus epidermidis (n = 32), Staphylococcus haemolyticus (n = 2) and Staphylococcus warneri (n = 8). The MIC of cefazolin was ≤2 mg/L for 30% of isolates and 75% had an MIC of ≤8 mg/L (MIC90 = 16 mg/L). This contrasted with MIC90s of cefoxitin, oxacillin and flucloxacillin, which were all ≥32 mg/L. WGS found a number of S. capitis isolates closely related to the globally established NRCS-A clone. CoNS displayed distinctly lower MIC values of cefazolin than of other agents tested. MIC variation may be related to binding affinity of PBP2a or regulation of expression of mecA by mecR1-mecI functional genes. Further, NRCS-A S. capitis strains were present in this Australian NICU before and after the unit underwent physical relocation, which raised questions about a common environmental source. It is considered justified to conduct a randomized clinical trial that assesses cefazolin versus vancomycin for management of late-onset neonatal sepsis.

To determine antimicrobial susceptibility and biofilm production among coagulase negative staphylococci at a tertiary care hospital

CoNS are gaining importance due to increase in resistance rates to betalactam antibiotics and multi drug resistance. Although specific virulence factors are not as clearly established, it seems clear that factors such as bacterial polysaccharide components, and ability to form biofilm are involved in attachment and/or persistence of bacteria on foreign materials. Biofilms usually result in persistent infections that cannot be easily resolved with standard antibiotic treatments; therefore, the biofilm formation ability and the resistance to antimicrobial therapy can be intimately related. A prospective cross-sectional study was done on purely isolated CoNS from various clinical samples from both out patients and inpatients. All the test strains were subjected to antimicrobial susceptibility testing. The ability to produce biofilm was detected by tube adherence method. Among 193 CoNS isolates 156 were from inpatients and 37 were from out patients. Methicillin resistant was seen in 80.31%. Of the total, 40.41% showed moderate biofilm formation by tube adherence method. 23.32% of isolates did not form biofilm. All the isolates from blood samples showed moderate (20/26) and strong (6/26) biofilm formation. Among non biofilm producers 66.67% were MS CoNS isolates and 33.33% were MRCoNS. 94.59% of biofilm producers were MRCoNS and 5.41% were MSCoNS. Production of biofilm was relatively more (1.16) among CoNS isolates of IPD than OPD. As Coagulase negative Staphylocooci are exhibiting multi drug resistance and are able to form biofilm, these organisms causing a major challenge for the physicians. Hence, such problems can be prevented by detection of biofilm producers and appropriate antibiotic doses modification. The issue of antibiotic resistance among CoNS needs to be addressed through a more rational use of existing antibiotics as well as the development of new antimicrobial agents.

Distribution of MR-CoNS in the clinical staffs of tertiary care hospital

Methicillin-resistant coagulase-negative staphylococci (MR-CoNS) associated infection is a growing concern in healthcare settings now a day. MR-CoNS are the main infectious agents of the hospital acquired infection. Clinical staffs to patients transmission of resistant strains have caused a rapid increase in the prevalence of antimicrobial resistance in recent years. Growing rate of antimicrobial resistant against available antibiotics of MR-CoNS is a developing problem in low income or lower middle income counties. This study was conducted to determine the occurrence MR-CoNS isolated from different clinical staffs of tertiary care hospital. This prospective study conducted in clinical staffs, nasal swab were collected from all the participants. Screening of CoNS were done on the basis of cultural, morphological and biochemical tests, identification and AST analysis done by VITEK-2 automated system. Methicillin resistance pattern was checked by VITEK-2 and Kirby-Bauer disc diffusion method according to CLSI guideline. A total of 129 nasal swab samples were collected from clinical staffs, of which n=81 isolates (85.6%) were CoNS. Among n=81 CoNS, (48.12%) , (41.97%) , (7.4%) and (2.4%) were reported. Out of n=81 CoNS isolates, n=26 were conformed as MR-CoNS. Maximum methicillin resistance were reported in 53.48% (14/26), 42.30% (11/26), 3.84% (1/26) and 0% (0/26). Theoccurrence rate of MR-CoNS are higher (20.6%) in the healthcare workers and most of the methicillin resistant-CoNS isolates shows high level of resistance against widely used antibiotics but all the isolates susceptible against vancomycin.

Identification of the Multiresistance Gene poxtA in Oxazolidinone-Susceptible Staphylococcus haemolyticus and Staphylococcus saprophyticus of Pig and Feed Origins.

Previous studies on the prevalence and transmission mechanism of oxazolidinone resistance gene poxtA in CoNS are lacking, which this study addresses. By screening 763 CoNS isolates from different sources of several livestock farms in Guangdong, China, 2018–2020, we identified that the poxtA was present in seven CoNS isolates of pig and feed origins. Species identification and multilocus sequence typing (MLST) confirmed that seven poxtA-positive CoNS isolates were composed of five ST64-Staphylococcus haemolyticus and two Staphylococcus saprophyticus isolates. All poxtA-positive Staphylococcus haemolyticus isolates shared similar pulsed-field gel electrophoresis (PFGE) patterns. Transformation assays demonstrated all poxtA-positive isolates were able to transfer poxtA gene to Staphylococcus aureus RN4220. S1-PFGE and whole-genome sequencing (WGS) revealed the presence of poxtA-carrying plasmids in size around 54.7 kb. The plasmid pY80 was 55,758 bp in size and harbored the heavy metal resistance gene czcD and antimicrobial resistance genes, poxtA, aadD, fexB and tet(L). The regions (IS1216E-poxtA-IS1216E) in plasmid pY80 were identified in Staphylococcus spp. and Enterococcus spp. with different genetic and source backgrounds. In conclusion, this was the first report about the poxtA gene in Staphylococcus haemolyticus and Staphylococcus saprophyticus, and IS1216 may play an important role in the dissemination of poxtA among different Gram-positive bacteria.

Ocular pathogens and antibiotic resistance in microbial keratitis over three years in Harbin, Northeast China.

PurposeUnderstanding the spectrum of ocular pathogens in a given geographic region is important for devising appropriate practical treatment. Therefore, we examined the pathogen spectrum and antibiotic resistance of microbial keratitis in northeast China.MethodsIn this retrospective observational study, we reviewed the microbiology laboratory records of patients diagnosed with microbial keratitis in a tertiary eye hospital in Harbin, northeast China, between 2017 and 2019, and analysed the pathogen spectrum and antibiotic susceptibility.ResultsWe collected 462 specimens, of which 282 exhibited positive cultures. Among these cultures, 257 were bacterial and 25 were fungal. Of the 257 bacterial isolates, 214 (83.27%) were gram positive whereas 43 (16.73%) were gram negative. The most prevalent gram‐positive pathogen was coagulase‐negative staphylococcus (CoNS; 58.37%), followed by Staphylococcus aureus (S. aureus; 20.62%) and Streptococcus pneumoniae (2.33%). Of the gram‐negative bacterial isolates, 10 were Pseudomonas aeruginosa (3.89%). The most frequently detected ocular pathogens from fungal isolates were Fusarium species (40%). We also found more culture‐positive cases of bacterial keratitis in summer. Overall, 16.98% S. aureus and 64.00% CoNS isolates were methicillin resistant. These methicillin‐resistant bacteria were also more likely to be resistant to other antibiotics, with multidrug resistance found in 77.78% methicillin‐resistant S. aureus and 90.63% methicillin‐resistant CoNS. However, all gram‐positive isolates were sensitive to vancomycin and linezolid.ConclusionCoagulase‐negative staphylococcus are the most common ocular pathogens in northeast China. We also show the prevalence of methicillin resistance and concurrent multidrug resistance among staphylococcal isolates. Monitoring the patterns of antimicrobial resistance could help in the management selection.

Prevalence and antimicrobial resistance patterns of nosocomial pathogens causing Surgical Site Infections in an Egyptian University hospital.

Background: Surgical site infections (SSI) are a common type of health care associated infections. The emergence of multidrug resistant nosocomial pathogens represents a major heath burden. This study was conducted to determine the frequency of isolation and patterns of antimicrobial resistance of nosocomial pathogens causing SSI in Zagazig University Hospitals. Methodology: Samples obtained from the infected surgical wounds were subjected to microbiological identification and antibiotic susceptibility testing. The role of extended spectrum beta lactamase (ESBL) and carbapenmases in bacterial resistance to some antibiotic were evaluated. Results: The most frequently isolated species were S. aureus (31%) followed by Klebseilla pneumoniae (22%), E. coli (15%), P. aeruginosa (11%), Coagulase negative staphylococci ( CoNs) (8%) , Proteus spp (7%) and Acinetobacter spp (6%). Methecillin resistance was detected in 38 (97 %) and 8 (80 %) S. aurues and CoNs isolates, respectively. Among Gram negative organisms, 65.8% of isolates were ESBL producers, of which 60% were Carbapenem resistant. Metallo-β-lactamase was detected in 30% of Gram negative isolates. Multi-drug resistance (MDR) was observed in 50 isolates (68.5%) where extensively drug resistance (XDR) occurred in 23 (31.5%) of Gram negative isolates. Conclusion: Most of Gram-negative isolates were MDR or XDR. Antibiotic therapy of SSI must be guided by microbiological culture and antibiotic sensitivity testing. Infection prevention and control practices need more improvement. Rationalisation of antibiotic prescription must be carried out. Post discharge surveillance of SSI needs to be considered.

Prevalence and distribution of resistance and enterotoxins/enterotoxin-like genes in different clinical isolates of coagulase-negative Staphylococcus

BackgroundCoagulase-negative staphylococcus (CoNS) is considered to be the major reservoirs for genes facilitating the evolution of S. aureus as a successful pathogen. The present study aimed to determine the occurrence of genes conferring resistance to fluoroquinolone, determining of the prevalence of insertion sequence elements IS256, IS257 and different superantigens (SAgs) among CoNS isolates obtained from various clinical sources.Materials and methodsThe current study conducted on a total of the 91 CoNS species recovered from clinical specimens in Hamadan hospitals in western Iran in 2017–2019. The antimicrobial susceptibility testing was performed using disk diffusion method and the presence of the IS256 and IS257, genes conferring resistance to fluoroquinolone and enterotoxins/enterotoxin-like encoding genes were investigated by polymerase chain reaction (PCR) method.ResultsAmong genes encoding classic enterotoxins, sec was the most frequent which was carried by 48.4% of the 91 isolates, followed by seb in 27.5% of the isolates. None of the CoNS isolates was found to be positive to enterotoxin-like encoding genes. In 11(12%) of all isolates that were phenotypically resistant to levofloxacin, 9 isolates (81.8%) were positive for gyrB, 8 isolates (72.7%) were positive for gyrA, 8 isolates (72.7%) harbored grlB and 7 isolates (63.6%) were found to carry grlA. The IS256 and IS257 were identified in 31.8% and 74.7% of the isolates, respectively. The results of statistical analysis showed a significant association between the occurrence of staphylococcal enterotoxins (SEs) encoding genes and antimicrobial resistance.ConclusionAntimicrobial resistant determinants and SEs are co-present in clinical CoNS isolates that confer selective advantage for colonization and survival in hospital settings. The coexistence of insertion elements and antibiotic resistance indicate their role in pathogenesis and infectious diseases.

Insights into the epidemiology of methicillin-resistant coagulase-negative Staphylococci carriage in community-based drug users

BackgroundCoagulase-negative Staphylococci (CoNS) are opportunistic pathogens. Methicillin-resistant CoNS (MRCoNS) remain a major cause of nosocomial infections, but limited information is available in communities. We, therefore, aimed to investigate the epidemiology of CoNS nasal carriage, especially MRCoNS in community-based drug users. MethodsDrug users were included in the cross-sectional study from three communities in Guangzhou, China. A face-to-face questionnaire was used to collect individual information and nasal swabs were collected to identify CoNS and MRCoNS isolates. Phenotypic and genotypic characterization of MRCoNS isolates were detected by using the disk diffusion method and polymerase chain reaction assays. Factors impacting MRCoNS carriage were assessed and estimated odds ratios (ORs) and 95% confidence intervals (CIs) by using logistic regression models. ResultsOverall, 353 drug users were eligible and included in the study. The prevalence of CoNS and MRCoNS were 42.78% and 29.18%, respectively. Having a history of being in prison (aOR = 2.16, 95% CI: 1.10−4.22) and a history of heroin snorting in the past 6 months (aOR = 1.87, 95% CI: 1.14−3.05) were risk factors for MRCoNS nasal carriage in drug users. The proportions of multidrug resistance for CoNS and MRCoNS isolates were respectively 54.38% and 66.07%. The predominant multidrug resistance pattern for CoNS and MRCoNS isolates was simultaneously non-susceptible to penicillin, cefoxitin and trimethoprim-sulfamethoxazole. MRCoNS isolates that were positive for erythromycin- (χ2 = 15.49, P < 0.001), tetracycline- (χ2 = 44.88, P < 0.001), penicillin- (χ2 = 3.86, P = 0.049), clindamycin- (χ2 = 4.18, P = 0.028), and gentamicin- (χ2 = 20.66, P < 0.001) resistance genes had significantly higher rates of resistance to corresponding antibiotics. ConclusionsThe prevalence of MRCoNS nasal carriage was high in community-based drug users. Drug users with risk factors should be paid greater attention to. The use of antibiotics in drug users needs further investigation and control.

Biofilm Formation Ability and Presence of Adhesion Genes among Coagulase-Negative and Coagulase-Positive Staphylococci Isolates from Raw Cow's Milk.

The capacity for biofilm formation is one of the crucial factors of staphylococcal virulence. The occurrence of biofilm-forming staphylococci in raw milk may result in disturbances in technological processes in dairy factories as well as the contamination of finished food products. Therefore, this study aimed to determine the prevalence and characteristics of staphylococcal biofilm formation in raw milk samples and to explore the genetic background associated with biofilm formation in those isolates. The material subjected to testing included 30 cow’s milk samples acquired from farms in the central part of Poland. A total of 54 staphylococcal strains were isolated from the samples, of which 42 were classified as coagulase-negative (CoNS) staphylococci belonging to the following species: S. haemolyticus, S. simulans, S. warneri, S. chromogenes, S. hominis, S. sciuri, S. capitis, S. xylosus and S. saprophyticus, while 12 were classified as S. aureus. The study examined the isolates’ capacity for biofilm formation and the staphylococcal capacity for slime production and determined the presence of genetic determinants responsible for biofilm formation, i.e., the icaA, icaD, bap and eno and, additionally, among coagulase-negative staphylococci, i.e., the aap, bhp, fbe, embP and atlE. Each tested isolate exhibited the capacity for biofilm formation, of which most of them (79.6%) were capable of forming a strong biofilm, while 5.6% formed a moderate biofilm, and 14.8% a weak biofilm. A capacity for slime production was demonstrated in 51.9% isolates. Most of the tested staphylococcal strains (90.7%) had at least one of the tested genes. Nearly half (47.6%) of the CoNS had the eno gene, while for S. aureus, the eno gene was demonstrated in 58.3% isolates. The frequency of the bap gene occurrence was 23.8% and 25% in CoNS strains and S. aureus, respectively. The fbe gene was demonstrated in only three CoNS isolates. The presence of the icaA was only demonstrated in CoNS strains (24.1%), while the icaD was found in both CoNS strains (21.4%) and S. aureus (100%). Among the CoNS, the presence of the embP (16.7%), aap (28.6%) and atlE (23.8%) was demonstrated as well. The obtained study results indicate that bacteria of the Staphylococcus spp. genus have a strong potential to form a biofilm, which may pose a hazard to consumer health.