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Background Methods Results Conclusion Chronic rhinosinusitis (CRS) is a persistent inflammatory condition of the sinus mucosa. While Staphylococcus aureus has been shown to play a significant role in mucosal barrier disruption in CRS patients, coagulase‐negative staphylococci (CoNS) such as Staphylococcus epidermidis and Staphylococcus lugdunensis are also implicated in CRS pathophysiology. This study investigates the effects of exoproteins secreted by planktonic and biofilm forms of clinical isolates of S. epidermidis and S. lugdunensis on the nasal epithelial barrier.Thirty‐one clinical isolates of CoNS were grown in planktonic and biofilm forms, and their exoproteins were concentrated. The epithelial barrier structure was assessed by measuring transepithelial electrical resistance (TEER) and the permeability of fluorescein isothiocyanate‐dextran. Toxicity and inflammatory response were also studied.Our findings demonstrate that exoproteins from all planktonic forms of S. lugdunensis disrupted the mucosal barrier, whereas only nine of 16 biofilm‐derived exoproteins had similar effects. Conversely, 11 of 15 exoproteins from planktonic S. epidermidis significantly disrupted barrier integrity; however, biofilm exoproteins did not. The study also showed that some exoproteins from planktonic S. epidermidis significantly reduced cell viability, while exoproteins from planktonic and biofilm forms of S. lugdunensis and biofilm S. epidermidis did not induce any statistically significant change in cell viability. Notably, four of 16 biofilm exoproteins from S. lugdunensis induced higher interleukin‐6 (IL‐6) secretion, whereas none of the S. epidermidis isolates showed a significant increase in IL‐6 secretion.Our results suggest that CoNS exoproteins may contribute to CRS etiopathogenesis. [ABSTRACT FROM AUTHOR]

Objective: We aimed to determine the demographic data, mortality, and morbidity of early- and late-neonatal sepsis cases, the etiologic agents in these cases, and the antibiotic susceptibility of these agents. Methods: This study was conducted retrospectively in a tertiary neonatal intensive care unit (NICU). The demographic, clinical, and laboratory data of newborns diagnosed with culture-proven sepsis within 24 months were evaluated. Results: Two hundred and eleven culture data points belonging to 197 infants were evaluated. Forty percent of the infants had a history of premature birth. The most common clinical findings were respiratory distress and feeding intolerance. Coagulase-negative staphylococci (CoNS) were detected most frequently as early- and late-sepsis agents. The most common Gram-negative bacteria detected as late-sepsis agents were Klebsiella spp. and Escherichia coli (E. coli). The overall mortality rate was 10%. Conclusions: Neonatal sepsis continues to have high mortality rates in tertiary NICUs. CoNS was the most common agent, highlighting the importance of developing and maintaining personnel training and handwashing practices. It will be important to consider the resistance rates of Klebsiella spp., the most common Gram-negative agent in late-onset sepsis (LOS) cases, to commonly used antibiotics in empirical treatments. [ABSTRACT FROM AUTHOR]

Objective: To determine the antibiotic resistance patterns and resistance genes in enterococci and coagulase-negative staphylococci (CoNS) isolated from poultry farms and nearby wells in Ibadan, Nigeria. Methods: Poultry litter and well water samples were collected from three farms. Isolation of bacterial isolates were done using bile esculin azide agar and presumptive identification was done using conventional methods and confirmed by 16S rRNA using PCR. Antibiotic susceptibility testing was by disk diffusion method, while antibiotics resistance genes (ARGs) was detected by PCR. Results: Of the 86 Gram-positive cocci isolated, 71 (82.6%) were CoNS comprising Staphylococcus (S.) sciuri (69.0%) and S. xylosus (31.0%), while the remaining 15 (17.4%) were Enterococcus spp. comprising Enterococcus (E.) faecalis (60.0%) and E. faecium (40.0%). Antibiotics resistance of the isolates showed that 91.5% (CoNS) and 93.0% (enterococci) were resistant to penicillin, 90.1% and 86.7% to tetracycline and 94.4% and 66.7% to erythromycin, respectively. Also, all the 15 (100%) Enterococcus spp. and 68 (95.8%) CoNS were multidrug resistant. The ARGs detected in the CoNS include: tetM (45.4%), tetL (9.0%) and ermB (27.3%), while tetM (9.1%), tetL (4.5%) and ermB (4.5%) were found in Enterococcus spp. Conclusions: This study revealed that poultry litters in the study area are pools of antimicrobial resistant enterococci and CoNS, as well as, the corresponding ARGs. Therefore, necessary monitoring and ample treatment of poultry waste should be done before their release into the environment.

The aim of this study was to obtain excellent probiotic bacteria from traditional Chinese fermented meat products. A total of 62 bacterial strains from the Miao, Dong and Buyi ethnic groups’ traditional fermented sour meat in selected regions of Guizhou, China were isolated, identified and evaluated for their probiotic potential. Twelve strains with good tolerance to simulated gastric juice were identified, including one strain of Companilactobacillus farciminis, five strains of Lactiplantibacillus plantarum, three strains of Staphylococcus saprophyticus and three strains of S. casei. Among them, C. farciminis KLMM196, L. plantarum KLMM198, S. saprophyticus KLMM253, S. casei KLMM307 and S. casei KLMM313 exhibited good gastrointestinal tolerance, hydrophobicity, self-aggregation ability, strong probiotic potential, and no hemolytic activity.

Non-aureus staphylococci and mammaliicocci (NASM) are the microorganisms most frequently isolated from milk. Given their numerosity and complexity, MALDI-TOF MS is one of the preferred species identification approaches. Nevertheless, reference mass spectra for the novel species Staphylococcus borealis were included only recently in the Bruker Biotyper System (MBT) library, and other species of veterinary interest such as S. rostri are still absent. This work provides an updated picture of the NASM species found in milk, gained by retrospectively analyzing the data relating to 21,864 milk samples, of which 6,278 from clinical mastitis (CM), 4,039 from subclinical mastitis (SCM), and 11,547 from herd survey (HS), with a spectrum library including both species. As a result, S. borealis was the second most frequently isolated NASM (17.07%) after S. chromogenes (39.38%) in all sample types, with a slightly higher percentage in CM (21.84%), followed by SCM (17.65%), and HS (14.38%). S. rostri was also present in all sample types (3.34%), reaching 8.43% of all NASM in SCM and showing a significant association (p < 0.01) with this condition. Based on our findings, the presence of S. borealis and S. rostri in milk and their potential association with mastitis might have been overlooked, possibly due to the difficulties in differentiating these species from other closely related NASM. Our results indicate that S. borealis could be a more frequent contributor to bovine udder infections than previously thought and that S. rostri should also not be underestimated considering its significant association with SCM. [ABSTRACT FROM AUTHOR]

Periprosthetic joint infections (PJI) of the shoulder are a devastating complication of shoulder arthroplasty and are commonly caused by Staphylococcus and Cutibacterium acnes. Absorbable calcium sulfate (CS) beads are sometimes used for delivering antibiotics in PJI. This study evaluates the in vitro effect of different combinations of gentamicin, vancomycin, and ertapenem in beads made from CS cement on the growth of C acnes and coagulase-negative Staphylococcus (CNS) strains. Three strains of C acnes and 5 strains of CNS from clinically proven shoulder PJI were cultured and plated with CS beads containing combinations of vancomycin, gentamicin, and ertapenem. Plates with C acnes were incubated anaerobically while plates with Staphylococcus were incubated aerobically at 37 °C. Zones of inhibition were measured at intervals of 3 and 7 days using a modified Kirby Bauer technique, and beads were moved to plates containing freshly streaked bacteria every seventh day. This process was run in triplicate over the course of 56 days. Statistical analysis was conducted using SPSS v. 28 with repeated measures analysis of variance (ANOVA) and pairwise comparisons with Tukey correction. In experiments with C acnes , beads containing ertapenem + vancomycin and vancomycin alone formed the largest zones of inhibition over time (P <.001). In experiments with Staphylococcus , beads containing vancomycin alone formed the largest zones of inhibition over time for all 5 strains (P <.001). Zones of inhibition were 1.4x larger for C acnes than for Staphylococcus with beads containing vancomycin alone. For both C acnes and Staphylococcus , beads containing ertapenem had the strongest initial effect, preventing all bacterial growth in C acnes and almost all growth for Staphylococcus during the first week but dropping substantially by the second week. Beads containing gentamicin alone consistently created smaller zones of inhibition than beads containing vancomycin alone, with vancomycin producing zones 5.3x larger than gentamicin in C acnes and 1.3x larger in Staphylococcus (P <.001). These data suggest that for both C acnes and Staphylococcal species, CS beads impregnated with vancomycin were most effective at producing a robust antibiotic effect. Additionally, ertapenem may be a viable supplement in order to create a more potent initial antibiotic effect but is not as effective as vancomycin when used alone. Gentamicin alone was not effective in maintaining consistent and long-term antibiotic effects. These results indicate that amongst the antibiotics currently commercially available to be used with CS, vancomycin is consistently superior to gentamicin in the setting of C. acnes and CNS. [ABSTRACT FROM AUTHOR]

Background and Aim: Staphylococci, which inhabit skin and mucous membranes in humans and animals, are opportunistic pathogens. Coagulase-positive and coagulase-negative staphylococci (CoNS) are the two main groups. Clinical abscesses in rabbits often harbor Staphylococcus aureus and CoNS. This study estimated S. aureus and CoNS prevalence, resistance profiles, antimicrobial-resistant genes, and the accessory gene regulator (agr) group in rabbit clinical abscesses. Materials and Methods: Sixty-seven abscesses were gathered from 67 rabbits who visited Prasu-Arthorn Animal Hospital in Nakornpathom, Thailand, from January 2014 to October 2015. Thirty-four subcutaneous, 29 dental, 2 ocular, 1 mammary gland, and 1 uterine abscess were present. Conventional methods, including Gram staining, mannitol fermentation, hemolysis on blood agar, catalase testing, and coagulase production, identified and isolated S. aureus and CoNS from all abscesses. All S. aureus and CoNS isolates underwent antimicrobial susceptibility testing using the disk diffusion method. Polymerase chain reaction was used to detect the presence of blaZ, aacA-aphD, msrA, tetK, gyrA, grlA, dfrG, and cfr antimicrobial-resistant genes. Methicillin resistance was identified through the detection of a cefoxitin-resistant phenotype and the presence of mecA gene. Further investigation was conducted on the agr group of S. aureus isolates. Results: In 67 abscesses, we found 19 S. aureus isolates in 9 abscesses (13.43%) and 37 CoNS isolates in 18 abscesses (26.87%), both majorly located at subcutaneous sites. About 59.46% of CoNS isolates were methicillin-resistant compared to 5.26% of S. aureus isolates. Methicillin-resistant S. aureus (MRSA) and methicillin-resistant CoNS (MRCoNS) both displayed multidrug resistance (MDR). Both MRSA and MRCoNS expressed multiple antimicrobial resistance genes, including blaZ, aacA-aphD, gyrA, grlA, msrA, tetK, and dfrG. Approximately 73.68% of the agr groups were agr I, 15.79% were agr III, and 10.53% were agr II. Conclusion: This study found a high prevalence of MRCoNS with antimicrobial resistance and multiple antimicrobial-resistant genes in rabbits with clinical abscesses. The effectiveness of antibiotics against infections caused by such strains is a matter of concern. Owners should be educated about the importance of good hygiene practices and judicious antibiotic use to prevent widespread antimicrobial resistance.

Abstract Background Recently, linezolid-resistant staphylococci have become an emerging problem worldwide. Understanding the mechanisms of resistance, molecular epidemiology and transmission of linezolid-resistant CoNS in hospitals is very important. Methods The antimicrobial susceptibilities of all isolates were determined by the microdilution method. The resistance mechanisms and molecular characteristics of the strains were determined using whole-genome sequencing and PCR. Results All the strains were resistant to oxacillin and carried the mecA gene; 13 patients (36.1%) had prior linezolid exposure. Most S. epidermidis and S. hominis isolates were ST22 and ST1, respectively. MLST typing and evolutionary analysis indicated most linezolid-resistant CoNS strains were genetically related. In this study, we revealed that distinct CoNS strains have different mechanisms of linezolid resistance. Among ST22-type S. epidermidis, acquisition of the T2504A and C2534T mutations in the V domain of the 23 S rRNA gene, as well as mutations in the ribosomal proteins L3 (L101V, G152D, and D159Y) and L4 (N158S), were linked to the development of linezolid resistance. In S. cohnii isolates, cfr, S158Y and D159Y mutations in the ribosomal protein L3 were detected. Additionally, emergence of the G2576T mutation and the cfr gene were major causes of linezolid resistance in S. hominis isolates. The cfr gene, G2576T and C2104T mutations, M156T change in L3 protein, and I188S change in L4 protein were found in S. capitis isolates. Conclusion The emergence of linezolid-resistant CoNS in the environment is concerning because it involves clonal dissemination and frequently coexists with various drug resistance mechanisms.

Introduction: Clinical significance of coagulase-negative staphylococci (CoNS) has been gradually acknowledged in both healthcare and clinical research, but approaches for their precise discrimination at the species level remain scarce. The current study aimed to evaluate the association of CoNS with orthopedic infections, where accurate and prompt identification of etiology is crucial for appropriate diagnosis and treatment decision-making. Methods: A 16S rRNA-based quantitative PCR (qPCR) assay was developed for the detection of Staphylococcus genus and two panels of 3-plex qPCR assays for further differentiation of six CoNS species with remarkable clinical significance, including S. epidermidis, S. haemolyticus, S. simulans, S. hominis, S. capitis, and S. caprae. All the assays exhibited excellent analytical performance. ΔCq (quantification cycle) between 16S rRNA and CoNS species-specific targets was established to determine the primary CoNS. These methods were applied to detect CoNS in wound samples from orthopedic patients with and without infection. Results and discussion: Overall, CoNS were detected in 17.8% (21/118) of patients with clinically suspected infection and in 9.8% (12/123) of patients without any infection symptom (p < 0.05). Moreover, the association with infection was found to be bacterial quantity dependent. S. epidermidis was identified as the predominant species, followed by S. simulans, S. haemolyticus, and S. hominis. Male sex, open injury, trauma, and lower extremity were determined as risk factors for CoNS infections. CoNS-positive patients had significantly longer hospitalization duration (20  days (15, 33) versus 13  days (7, 22) for Staphylococcus-negative patients, p = 0.003), which could be a considerable burden for healthcare and individual patients. Considering the complex characteristics and devastating consequences of orthopedic infections, further expanding the detection scope for CoNS may be pursued to better understand the etiology of orthopedic infections and to improve therapeutic strategies. [ABSTRACT FROM AUTHOR]

The antibiotic resistance (ABR) crisis is an urgent global health priority. Staphylococci are among the problematic bacteria contributing to this emergency owing to their recalcitrance to many clinically important antibiotics. Staphylococcal pathogenesis is further complicated by the presence of small colony variants (SCVs), a bacterial subpopulation displaying atypical characteristics including retarded growth, prolific biofilm formation, heightened antibiotic tolerance, and enhanced intracellular persistence. These capabilities severely impede current chemotherapeutics, resulting in chronic infections, poor patient outcomes, and significant economic burden. Tackling ABR requires alternative measures beyond the conventional options that have dominated treatment regimens over the past 8 decades. Non-antibiotic therapies are gaining interest in this arena, including the use of honey, which despite having ancient therapeutic roots has now been reimagined as an alternative treatment beyond just traditional topical use, to include the treatment of an array of difficult-to-treat staphylococcal infections. This literature review focused on Manuka honey (MH) and its efficacy as an anti-staphylococcal treatment. We summarized the studies that have used this product and the technologies employed to study the antibacterial mechanisms that render MH a suitable agent for the management of problematic staphylococcal infections, including those involving staphylococcal SCVs. We also discussed the status of staphylococcal resistance development to MH and other factors that may impact its efficacy as an alternative therapy to help combat ABR. [ABSTRACT FROM AUTHOR]

Background and Aim: Staphylococci, which inhabit skin and mucous membranes in humans and animals, are opportunistic pathogens. Coagulase-positive and coagulase-negative staphylococci (CoNS) are the two main groups. Clinical abscesses in rabbits often harbor Staphylococcus aureus and CoNS. This study estimated S. aureus and CoNS prevalence, resistance profiles, antimicrobial-resistant genes, and the accessory gene regulator (agr) group in rabbit clinical abscesses. Materials and Methods: Sixty-seven abscesses were gathered from 67 rabbits who visited Prasu-Arthorn Animal Hospital in Nakornpathom, Thailand, from January 2014 to October 2015. Thirty-four subcutaneous, 29 dental, 2 ocular, 1 mammary gland, and 1 uterine abscess were present. Conventional methods, including Gram staining, mannitol fermentation, hemolysis on blood agar, catalase testing, and coagulase production, identified and isolated S. aureus and CoNS from all abscesses. All S. aureus and CoNS isolates underwent antimicrobial susceptibility testing using the disk diffusion method. Polymerase chain reaction was used to detect the presence of blaZ, aacA-aphD, msrA, tetK, gyrA, grlA, dfrG, and cfr antimicrobial-resistant genes. Methicillin resistance was identified through the detection of a cefoxitin-resistant phenotype and the presence of mecA gene. Further investigation was conducted on the agr group of S. aureus isolates. Results: In 67 abscesses, we found 19 S. aureus isolates in 9 abscesses (13.43%) and 37 CoNS isolates in 18 abscesses (26.87%), both majorly located at subcutaneous sites. About 59.46% of CoNS isolates were methicillin-resistant compared to 5.26% of S. aureus isolates. Methicillin-resistant S. aureus (MRSA) and methicillin-resistant CoNS (MRCoNS) both displayed multidrug resistance (MDR). Both MRSA and MRCoNS expressed multiple antimicrobial resistance genes, including blaZ, aacA-aphD, gyrA, grlA, msrA, tetK, and dfrG. Approximately 73.68% of the agr groups were agr I, 15.79% were agr III, and 10.53% were agr II. Conclusion: This study found a high prevalence of MRCoNS with antimicrobial resistance and multiple antimicrobialresistant genes in rabbits with clinical abscesses. The effectiveness of antibiotics against infections caused by such strains is a matter of concern. Owners should be educated about the importance of good hygiene practices and judicious antibiotic use to prevent widespread antimicrobial resistance. [ABSTRACT FROM AUTHOR]

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Purpose: To assess Gram-positive bacterial (GPB) bloodstream infection (BSI) in neonates, covering incidence, morbidity, mortality, antimicrobial resistance patterns and biomarkers in Region Stockholm, Sweden between 2006 and 2016. Methods: A population-based retrospective epidemiological study including infants with GPB-BSI, admitted to the neonatal units at Karolinska University Hospital (KUH). Data were collected from patient records, the Swedish Neonatal Quality Register, the microbiological laboratory at KUH and the Swedish Public Health Agency. Results: We identified 357 infants with GPB-BSI, representing an incidence of 1.47/1000 live births (LB). Group B streptococcus (GBS) was the most common pathogen causing BSI in full-term infants and early-onset sepsis (EOS) (0.20/1000 LB), while coagulase-negative staphylococci (CoNS) were predominant in infants born very preterm and in late-onset sepsis (LOS) (0.79/1000 LB). There were no fatal GBS BSI cases, but 10.2% developed meningitis. The GPB case fatality rate was 9.5% and the sepsis fatality rate 2.8%. In GPB-BSI, 1/10 did not have an elevated C-reactive protein level. Staphylococcus aureus (S. aureus) BSI increased during the study period, but no methicillin or vancomycin resistant strains were found. The antimicrobial resistance (AMR) rate was highest in CoNS isolates. Conclusion: GPB-BSI was four times more common than Gram-negative BSI in neonates but resulted in lower mortality rate. GBS was the most common pathogen in full-term infants and in EOS. CoNS was the most common pathogen in LOS and infants born very preterm, and the AMR rate was high in these isolates. The increasing trend of S. aureus BSI indicates a need of further investigation. [ABSTRACT FROM AUTHOR]

Background: Oral cavity is an ecological niche for colonization of staphylococci, which are a major bacterial species causing community-acquired infections in humans. In this study, prevalence, and characteristics of staphylococci in oral cavity and skin of healthy individuals were investigated in northern Japan. Methods: Saliva from oral cavity and swab from skin surface of hand were collected and cultured on selective media. Species of the isolates were identified genetically, and ST was determined for S. aureus and S. argenteus. Genes associated with antimicrobial resistance were detected by PCR. Results: Among 166 participants, a total of 75 S. aureus isolates were obtained from 61 individuals (37 %), and recovered more frequently in oral cavity (n = 48) than skin (n = 27). Among 23 STs identified in S. aureus isolates, ST8 (CC8), ST15 (CC15), and ST188 (CC1) were the most common (10 isolates each), with STs of CC1 being dominant (17 isolates). Methicillin-resistant S. aureus (MRSA) was isolated in the skin of two individuals and belonged to ST1 and ST6. Resistance to erythromycin and gentamicin associated with erm(A) and aac(6’)-Ie-aph(2”)-Ia, respectively, was more commonly found in ST5 and ST8 isolates. One S. argenteus isolate (ST2250, mecA-negative) was recovered from oral cavity of a participant (0.6 %). A total of 186 isolates of coagulase-negative staphylococci (CoNS) were recovered from 102 participants and identified into 14 species, with S. warneri being the most common (n = 52), followed by S. capitis (n = 42), S. saprophyticus (n = 20) and S. haemolyticus (n = 19). mecA was detected in S. saprophyticus, S. haemolyticus, and S. caprae, while arginine-catabolic mobile element (ACME) in only S. capitis and S. epidermidis. Conclusion: S. aureus was more prevalent in oral cavity than skin surface, belonging to three major STs, with CC1 being a dominant lineage. The prevalence of antimicrobial resistance was distinct depending on CoNS species.

This study investigates the community of coagulase-negative staphylococci (CNS) in samples of Sjenica sheep's ham from different villages across three seasons over a 120-day maturation period. The CNS group was identified into five distinct species: Staphylococcus epidermidis, S. saprophyticus, S. carnosus, S. xylosus, and S. equorum. The safety evaluation revealed no hemolytic activity in any tested isolates, while resistance to novobiocin was observed in S. xylosus and S. saprophyticus isolates. The technological properties of the isolated CNS were assessed, including tolerance to different temperatures, pH values, salt concentrations, as well as proteolytic and lipolytic activity. Results demonstrated the ability of all tested isolates to grow across various pH values and salt concentrations, with isolates of S. carnosus and S. epidermidis showing tolerance to 45°C. The best technological properties were shown by S. carnosus, followed by S. equorum. These findings provide insights into the potential applications of CNS in food processing industries, as putative starters.

Objective Fluorine-18 fluorodeoxyglucose (18 F-FDG) positron emission tomography/computed tomography (PET/CT) has gained attention as an emerging tool in case of suspicion of infection on spine, whether native or instrumented. However, the diagnostic performance of 18 F-FDG PET/CT in clinically occult low-grade surgical site infection (SSI) after spinal fusion, an important risk factor for pseudarthrosis, remains unknown. Methods We retrospectively identified all the presumed aseptic patients with pseudarthrosis confirmed by revision surgery who underwent preoperative 18 F-FDG PET/CT scans performed between April 2019 and November 2022. These patients were presumed aseptic because they did not have clinical signs or laboratory tests suggestive of SSI, preoperatively. The PET/CT images were analyzed in consensus by two nuclear medicine physicians blinded to the clinical, biological, and imaging information. Visual assessment of increased uptake around cage/intervertebral disk space (and/or hardware) higher than background recorded from the first normal adjacent vertebra was interpreted as positive. Image data were also quantitatively analyzed by the maximum standardized uptake value as an index of 18 F-FDG uptake, and the ratio between the uptake around cage/intervertebral disk space (and/or hardware) and background recorded from the first normal adjacent vertebra was calculated. The final diagnosis of infection was based on intraoperative cultures obtained during pseudarthrosis revision surgery. Results Thirty-six presumed aseptic patients with surgically confirmed pseudarthrosis after spinal fusion underwent preoperative 18 F-FDG PET/CT scans. Cultures of samples from revisions found that 20 patients (56%) were infected. The most frequent isolated bacterium was Cutibacterium acnes (C. acnes) in 15 patients (75%), followed by coagulase-negative staphylococci (CNS) in 7 patients (33%). Two patients had co-infections involving both C. acnes and CNS. Of the 36 PET/CT studied in this study, 12 scans were true-negative, 10 true-positive, 10 false-negative, and 4 false-positive. This resulted in sensitivity, specificity, positive predictive value, negative predictive value, and diagnostic accuracy of 50%, 75%, 71%, 55%, and 61%, respectively. Conclusion In presumed aseptic pseudarthrosis after spinal fusion, 18 F-FDG PET/CT offers good specificity (75%) but low sensitivity (50%) to identify occult SSI. The high prevalence (56%) of SSI, mostly caused by C. acnes (75%), found in our presumed aseptic cohort of patients supports the utility of systematic intraoperative cultures in revision cases for pseudarthrosis. [ABSTRACT FROM AUTHOR]

Ripened sheep sausages are widely consumed in Italy, particularly in Sardinia. Despite their driving role in flavor and color development, coagulase-negative staphylococci in these products have been rarely investigated. A total of 70 CoNS cultures isolated from Sardinian sheep sausages were characterized by rep-PCR and M13-RAPD typing and identified by 16S rDNA sequencing. S. xylosus and S. equorum accounted for more than 70% of the total isolates, whilst S. pasteuri (8.5%), S. succinus (2.8%), and S. haemolyticus (2.8%) were less represented. The genes encoding the synthesis of putrescine, tyramine, cadaverine, and histamine were evaluated by PCR. None of the strains hosted genes for decarboxylases, except one S. pasteuri strain that was potentially a tyramine-producer. Antibiotic resistance was evaluated, along with nitrate reductase, lipolytic, and proteolytic activity, in a pool of selected cultures. Resistance to the primary antibiotics was rather widespread. S. xylosus, S. equorum, and S. pasteuri strains were all resistant to amoxicillin and kanamycin. S. equorum strains were sensitive to all tested antibiotics. S. xylosus strains were all resistant to penicillin B. Conversely, all S. pasteuri strains were resistant to both ampicillin and penicillin B, and four out of five strains exhibited tetracycline resistance. The high variability in the production of sheep sausages makes the search for adjunct cultures of crucial relevance. According to this perspective, the characterization of the autochthonous CSN population represents the first step to approach a starter selection. [ABSTRACT FROM AUTHOR]

Objectives: Little is known about efficacy and safety of ethanol lock therapy (ELT) to treat totally implantable venous access device (TIVAD) infections. The objective of this trial was to evaluate the effectiveness and safety profile of a local treatment with ELT without removal for TIVAD infection due to coagulase-negative staphylococci. Methods: We performed a prospective, multicenter, double-blind, randomized clinical trial comparing the efficacy of 40% ELT versus vancomycin lock therapy (VLT) in TIVAD infections due to coagulase-negative staphylococci, complicated or not by bloodstream infection. Results: Thirty-one patients were assigned to the ELT group and 30 to the VLT arm. Concomitant bacteremia was present in 41 patients (67.2%). Treatment success was 58.1 % (18 of 31) for the ELT arm and 46.7% (14 of 30) for the VLT arm (p = 0.37). The overall treatment success was 52.5% (32). The risk of treatment failure due to uncontrolled infections, superinfections, and mechanical complications did not differ significantly between participants receiving ELT (13 out of 31 [42%]) and those receiving VLT (16 out of 30 [53%]) with a hazard ratio of 0.70 (p = 0.343; 95% CI [0.34–1.46], Cox model). Catheter malfunctions were significantly more frequent in the ELT arm (11 patients versus 2 in the VLT group, p = 0.01). Conclusions: We found an overall high rate of treatment failure that did not differ between the ELT arm and the VLT arm. TIVAD removal must be prioritized to prevent complications (uncontrolled infections, superinfections, and catheter malfunctions) except in exceptional situations. [ABSTRACT FROM AUTHOR]

Non-aureus staphylococci and mammaliicocci (NASM) are associated with bovine mastitis and increased milk somatic cell count (SCC) but their relationships with mammary gland health at the species level are not clearly defined. Regional differences have also been reported in their specific prevalence. The implementation of MALDI-TOF MS in milk microbiology is generating large and dependable datasets with the potential of providing useful epidemiological information. We present the retrospective analysis of 17,213 milk samples sent to our laboratory in 2021–2022, including 13,146 quarter samples from cows with subclinical (SCM) or clinical mastitis (CM) from 104 farms, and 4,067 composite herd survey (HS) samples from 21 farms. NASM were isolated from 21.12% of SCM, 11.49% of CM, and 15.59% of HS milk samples. The three most frequently identified NASM in SCM milk were Staphylococcus chromogenes (33.33%), S. haemolyticus (26.07%), and S. epidermidis (10.65%); together with S. microti and S. hyicus, these species were significantly more prevalent in quarters with SCM (p < 0.05). The three most frequently identified NASM in CM milk were S. chromogenes (31.69%), S. haemolyticus (21.42%), and Mammaliicoccus sciuri (18.38%), although no significant associations were found between these NASM species and CM. The three most frequently identified NASM in HS milk were S. chromogenes (44.49%), S. epidermidis (17.84%), and S. haemolyticus (17.23%), with S. chromogenes being isolated in all the farms sending HS milk (100%). In conclusion, this retrospective study provides the first information on the NASM species isolated from cow milk in Italy, expanding our knowledge on the epidemiology of NASM at the species level and providing further insights into their relationships with mammary gland health in modern dairy farms. [ABSTRACT FROM AUTHOR]

Staphylococcus capitis has been recognized as a relevant opportunistic pathogen, particularly its persistence in neonatal ICUs around the world. Therefore, the aim of this study was to describe the epidemiological profile of clinical isolates of S. capitis and to characterize the factors involved in the persistence and pathogenesis of these strains isolated from blood cultures collected in a hospital in the interior of the state of São Paulo, Brazil. A total of 141 S. capitis strains were submitted to detection of the mecA gene and SCCmec typing by multiplex PCR. Genes involved in biofilm production and genes encoding enterotoxins and hemolysins were detected by conventional PCR. Biofilm formation was evaluated by the polystyrene plate adherence test and phenotypic resistance was investigated by the disk diffusion method. Finally, pulsed-field gel electrophoresis (PFGE) was used to analyze the clonal relationship between isolates. The mecA gene was detected in 99 (70.2%) isolates, with this percentage reaching 100% in the neonatal ICU. SCCmec type III was the most prevalent type, detected in 31 (31.3%) isolates and co-occurrence of SCCmec was also observed. In vitro biofilm formation was detected in 46 (32.6%) isolates but was not correlated with the presence of the ica operon genes. Furthermore, biofilm production in ICU isolates was favored by hyperosmotic conditions, which are common in ICUs because of the frequent parenteral nutrition. Analysis of the clonal relationship between the isolates investigated in the present study confirms a homogeneous profile of S. capitis and the persistence of clones that are prevalent in the neonatal ICU and disseminated across the hospital. This study highlights the adaptation of isolates to specific hospital environments and their high clonality. [ABSTRACT FROM AUTHOR]

This study aimed to genotypic and phenotypic analyses of the enterotoxigenic potential of Staphylococcus spp. isolated from raw milk and raw milk cheeses. The presence of genes encoding staphylococcal enterotoxins (SEs), including the classical enterotoxins (sea-see), non-classical enterotoxins (seg-seu), exfoliative toxins (eta-etd) and toxic shock syndrome toxin-1 (tst-1) were investigated. Isolates positive for classical enterotoxin genes were then tested by SET-RPLA methods for toxin expression. Out of 75 Staphylococcus spp. (19 Staphylococcus aureus and 56 CoNS) isolates from raw milk (49/65.3%) and raw milk cheese samples (26/34.7%), the presence of enterotoxin genes was confirmed in 73 (97.3%) of them. Only one isolate from cheese sample (1.3%) was able to produce enterotoxin (SED). The presence of up to eight different genes encoding enterotoxins was determined simultaneously in the staphylococcal genome. The most common toxin gene combination was sek, eta present in fourteen isolates (18.7%). The tst-1 gene was present in each of the analyzed isolates from cheese samples (26/34.7%). Non-classical enterotoxins were much more frequently identified in the genome of staphylococcal isolates than classical SEs. The current research also showed that genes tagged in S. aureus were also identified in CoNS, and the total number of different genes detected in CoNS was seven times higher than in S. aureus. The obtained results indicate that, in many cases, the presence of a gene in Staphylococcus spp. is not synonymous with the ability of enterotoxins production. The differences in the number of isolates with genes encoding SEs and enterotoxin production may be mainly due to the limit of detection of the toxin production method used. This indicates the need to use high specificity and sensitivity methods for detecting enterotoxin in future studies. [ABSTRACT FROM AUTHOR]

The antibiotic resistance (ABR) crisis is an urgent global health priority. Staphylococci are among the problematic bacteria contributing to this emergency owing to their recalcitrance to many clinically important antibiotics. Staphylococcal pathogenesis is further complicated by the presence of small colony variants (SCVs), a bacterial subpopulation displaying atypical characteristics including retarded growth, prolific biofilm formation, heightened antibiotic tolerance, and enhanced intracellular persistence. These capabilities severely impede current chemotherapeutics, resulting in chronic infections, poor patient outcomes, and significant economic burden. Tackling ABR requires alternative measures beyond the conventional options that have dominated treatment regimens over the past 8 decades. Non-antibiotic therapies are gaining interest in this arena, including the use of honey, which despite having ancient therapeutic roots has now been reimagined as an alternative treatment beyond just traditional topical use, to include the treatment of an array of difficult-to-treat staphylococcal infections. This literature review focused on Manuka honey (MH) and its efficacy as an anti-staphylococcal treatment. We summarized the studies that have used this product and the technologies employed to study the antibacterial mechanisms that render MH a suitable agent for the management of problematic staphylococcal infections, including those involving staphylococcal SCVs. We also discussed the status of staphylococcal resistance development to MH and other factors that may impact its efficacy as an alternative therapy to help combat ABR.

ABSTRACTThe species- and clone-specific susceptibility of Staphylococcus cells for bacteriophages is governed by the structures and glycosylation patterns of wall teichoic acid (WTA) glycopolymers. The glycosylation-dependent phage-WTA interactions in the opportunistic pathogen Staphylococcus epidermidis and in other coagulase-negative staphylococci (CoNS) have remained unknown. We report a new S. epidermidis WTA glycosyltransferase TagE whose deletion confers resistance to siphoviruses such as ΦE72 but enables binding of otherwise unbound podoviruses. S. epidermidis glycerolphosphate WTA was found to be modified with glucose in a tagE-dependent manner. TagE is encoded together with the enzymes PgcA and GtaB providing uridine diphosphate-activated glucose. ΦE72 transduced several other CoNS species encoding TagE homologs, suggesting that WTA glycosylation via TagE is a frequent trait among CoNS that permits interspecies horizontal gene transfer. Our study unravels a crucial mechanism of phage-Staphylococcus interaction and horizontal gene transfer, and it will help in the design of anti-staphylococcal phage therapies.IMPORTANCEPhages are highly specific for certain bacterial hosts, and some can transduce DNA even across species boundaries. How phages recognize cognate host cells remains incompletely understood. Phages infecting members of the genus Staphylococcus bind to wall teichoic acid (WTA) glycopolymers with highly variable structures and glycosylation patterns. How WTA is glycosylated in the opportunistic pathogen Staphylococcus epidermidis and in other coagulase-negative staphylococci (CoNS) species has remained unknown. We describe that S. epidermidis glycosylates its WTA backbone with glucose, and we identify a cluster of three genes responsible for glucose activation and transfer to WTA. Their inactivation strongly alters phage susceptibility patterns, yielding resistance to siphoviruses but susceptibility to podoviruses. Many different CoNS species with related glycosylation genes can exchange DNA via siphovirus ΦE72, suggesting that glucose-modified WTA is crucial for interspecies horizontal gene transfer. Our finding will help to develop antibacterial phage therapies and unravel routes of genetic exchange.

Background and objectives: Coagulase-negative staphylococci (CoNS) are considered important causative agents of hospital acquired infection. These organisms are found in various clinical specimens from hospitalized patients. Present study was carried out to determine the species distribution and antimicrobial susceptibility pattern of CoNS isolated from clinical specimens at a tertiary care hospital. Materials and methods: CoNS isolated from various clinical samples were included in this study. The isolates were identified by colony morphology, Gram’s staining, catalase and coagulase tests. Further differentiation of species was performed by susceptibility to novobiocin, urease activity and ornithine decarboxylase test. Antibiotic susceptibility testing was performed according to the Clinical and Laboratory Standard Institute (CLSI) guidelines. Results: Total 108 isolates of CoNS were included and analysed. Out of 108 CoNS, S. epidermidis was the most common species (36.1%) followed by S. saprophyticus (23.1%), S. hemolyticus (17.6%), S. hominis (13%) and S. lugdunensis (10.2%). Most of the isolates showed resistance to penicillin, oxacillin, amoxycillin, erythromycin, ciprofloxacin and ofloxacin. All the isolates were sensitive to vancomycin. Conclusions: CoNS emerged as an important nosocomial pathogen and should not be neglected as contaminant. High rate of antimicrobial resistance warrants susceptibility testing prior to the treatment of CoNS. IMC J Med Sci. 2024; 18(1):010. DOI: https://doi.org/10.55010/imcjms.18.010 *Correspondence: Sabiha S Tamboli, Department of Microbiology, Parbhani Medical College and RP Hospital Research Institute, Pathri Road, Parbhani, Maharashtra, India. Email: sabihatamboli77@gmail.com

The study investigates the prevalence of Staphylococcus aureus and coagulasenegative staphylococci in raw milk and dairy products and assesses their toxin-related pathogenic potential and methicillin resistance. A total of 1015, raw milk (260) and dairy samples (325 cheeses, 180 yogurts, 140 ice creams, 110 butter samples) were collected and analyzed. The prevalence of Staphylococcus aureus and coagulase-negative staphylococci were 3.2% and 5.3% with mean counts of 3.46 and 3.16 log CFU/mL-g, respectively. Three (sea, seb, see) of five (sea, seb, sec, sed, see) staphylococcal enterotoxin (SE) genes, two (tss, etb) of four (tss, pvl, eta, etb) virulence-associated genes, and the absence of methicillin resistance (mecA) gene were defined by polymerase chain reaction. SE sea (6.9%), seb (2.3%) and see (1.1%) genes were detected in Staphylococcus aureus from one milk and seven different cheese samples. The presence of multiple enterotoxin genes (sea and see) was detected in a Staphylococcus aureus isolate from one cheese. However, the milk-sourced one coagulase-negative staphylococci possessed both the tss and etb virulence genes. The finding in this study indicates that the frequency of coagulase-negative staphylococci was higher than Staphylococcus aureus and moreover, toxin genes associated with human infections were assigned in coagulase-negative staphylococci while enterotoxin genes were determined among Staphylococcus aureus. In terms of food safety perspective, coagulasenegative staphylococci are ignored and they are not considered in standard food surveillance analysis. But the presence of virulent coagulase-negative staphylococci in foods is a public health concern. The results obtained from this study are significant as it demonstrates that pathogenic coagulase-negative staphylococci are found in foods, and provides data from Turkey. Additional research is required concerning coagulase-negative staphylococci in the food matrix and clinical isolates.

Abstract Background Sepsis in low-birth-weight neonates remains one of the most significant causes of neonatal morbidity and mortality. Approximately 3 million newborns suffer from sepsis globally every year. The aim of this study was to compare demographic and clinical features, as well as etiology and antibiotic susceptibility, of the main pathogens related to neonatal sepsis in two neonatal intensive units during a two-year period. Methods We observed early-onset (EO-BSI) and late-onset bloodstream infections (LO-BSI) cases in two high-reference neonatal intensive care units (NICU) over a 24-month period (2016–2017). Samples of patients’ blood were tested for the presence of the microorganisms. All bacterial isolates were tested for susceptibility to antibiotics. Results The majority of sepsis cases weighed above 1000 g and were born by cesarean section. About 10% of the EO-BSI group died. There were differences in the EO-BSI /LO-BSI ratio in the compared wards due to differences among the admitted children. The most common pathogens isolated from blood were coagulase-negative staphylococci (CoNS) were represented by two dominating species: S. epidermidis and S. haemolyticus, followed by Klebsiella spp. strains and E.coli, which were mostly found in EO-BSI cases. No single S. agalactiae (GBS) strain was isolated. The majority of CoNS strains were resistant to methicillin, half were resistant to aminoglycosides, and one-third were resistant to macrolides and lincosamides. Half of the Gram-negative rods were resistant to beta-lactams. Conclusions The epidemiology of sepsis in two observed NICUs is comparable to data obtained from other studies with a predominance of methicillin-resistant CoNS in LO-BSI and beta-lactam resistant E. coli in EO-BSI. It is of importance that the campaign for controlling GBS carriage in pregnant women in Poland resulted in the disappearance of GBS as a cause of sepsis. Unfortunately, there are no such measures to control E.coli related sepsis.

Objective: We aimed to determine the demographic data, mortality, and morbidity of early- and late-neonatal sepsis cases, the etiologic agents in these cases, and the antibiotic susceptibility of these agents. Methods: This study was conducted retrospectively in a tertiary neonatal intensive care unit (NICU). The demographic, clinical, and laboratory data of newborns diagnosed with culture-proven sepsis within 24 months were evaluated. Results: Two hundred and eleven culture data points belonging to 197 infants were evaluated. Forty percent of the infants had a history of premature birth. The most common clinical findings were respiratory distress and feeding intolerance. Coagulase-negative staphylococci (CoNS) were detected most frequently as early- and late-sepsis agents. The most common Gram-negative bacteria detected as late-sepsis agents were Klebsiella spp. and Escherichia coli (E. coli). The overall mortality rate was 10%. Conclusions: Neonatal sepsis continues to have high mortality rates in tertiary NICUs. CoNS was the most common agent, highlighting the importance of developing and maintaining personnel training and handwashing practices. It will be important to consider the resistance rates of Klebsiella spp., the most common Gram-negative agent in late-onset sepsis (LOS) cases, to commonly used antibiotics in empirical treatments.

Abstract Background Peritonitis is the most important complication of peritoneal dialysis (PD) and coagulase-negative staphylococci (CNS) are a frequent cause of dialysis-related infections. The association between SCCmec typing with psm-mec positivity in staphylococci and PD-related infections has not been identified. We aim to investigate the molecular epidemiology of CNS isolated from PD-peritonitis in a single Chinese center, focusing on the genetic determinants conferring methicillin resistance. Methods We collected 10 genetically unrelated CNS isolates from 10 patients with CNS PD-related peritonitis. The patients were divided into two groups based on the results of MIC to oxacillin: the methicillin-resistant CNS (MRCNS) and methicillin-sensitive CNS (MSCNS) groups. The biofilm formation group (BFG) and the non-biofilm formation group (NBFG) were used as the control groups. Phenotypic and molecular methods were used to analyze SCCmec types I, II and III, associated genes and biofilm formation and the existence of psm-mec. The demographic data and clinical indicators were collected. Results Ten CNS PD-related peritonitis patients were enrolled for this study. There were 6 MRCNS and 4 MRCNS isolates. SCCmec types were fully determined in 10 isolates. Seven staphylococci (70%) carried SCCmec, of which 4 isolates carried single SCCmec type I (40%) and 3 isolates had multiple SCCmec elements (I + III). Of the 6 MRCNS isolates, 3 carried SCCmec type I (50%) and 2 isolates carried SCCmec type I + III (33.3%). A high diversity of ccr types, mec complexes and ccr-mec complex combinations was identified among the 10 CNS isolates. The psm-mec gene was detected in 2/10 (20%) CNS isolates. There was no mutation in the psm-mec gene. Conclusions The majority of isolates were hospital-associated isolates. Furthermore, 2 psm-mec positive isolates were MRCNS in the NBFG. The PD patients frequent exposure to hospital would be the main risk factor. The presence of the psm-mec signal in the spectra of the MRCNS tested here demonstrates the presence of certain SCCmec cassettes that convey methicillin resistance.

The study reveals crucial information on S. capitis, a potentially dangerous bacterium that can cause sepsis in hospitalized adult patients, including its frequency and patterns of drug resistance which could significantly contribute to the existing body of knowledge in this field. This retrospective study was conducted in the King Fahad Medical City (KFMC), Riyadh, Saudi Arabia, from June 2019 to November 2022. The comprehensive data collection and analysis provide valuable information on the prevalence of S. capitis in different infection sites, antibiotic sensitivity profiles, and the association with patient demographics. A total of 219 S. capitis isolates from hospitalized patients with sepsis tested positive. Infection was most common in venous blood (139 patients, 63.5%), followed by central blood (24 patients, 11%). Other locations included venipuncture (48 patients, 21.9%) and cerebrospinal fluid (three patients, 1.4%). The results highlight the clinical significance of S. capitis as a major contributor to sepsis, especially in male patients, and emphasize the need for appropriate antibiotic selection for effective treatment. Furthermore, this study raises concerns about the high rates of methicillin resistance observed in S. capitis isolates, emphasizing the urgency in tackling antibiotic resistance and promoting judicious antibiotic prescription practices. Moreover, the identification of a decreased susceptibility to vancomycin as a potential explanation for its reduced effectiveness in treating S. capitis infections adds a crucial aspect to the discussion of treatment options and underscores the need for alternative therapeutic strategies.

Abstract Background Healthcare workers may pave the way for increased infections in hospitalized patients by coagulase-negative staphylococci (CoNS). Biofilm formation and antibiotic resistance are the major problems posed by CoNS in nosocomial infections. In this study, we determined biofilm production level and the distribution of biofilm-associated and virulence genes, including icaADBC, aap, bhp, atlE, embp, and fbe, as well as IS256, IS257, mecA, and ACME clusters (arc-A, opp-3AB) among 114 clinical (n = 57) and healthcare workers (n = 57) CoNS isolates in Kerman, Iran. Results In this study, more than 80% (n = 96) of isolates were methicillin-resistant CoNS (MR-CoNS). Out of 114 isolates, 33% (n = 38) were strong biofilm producers. Strong biofilm formation was found to be significantly different between clinical and healthcare workers’ isolates (P

The influence of the starter cultures, such as Lactobacillus rhamnosus, L. plantarum, Kocuria rosea, Staphylococcus carnosus, L. plantarum, L. rhamnosus and L. paracasei, on the functional-technological and physicochemical characteristics of the pork meat during the salting is investigated in this paper. It has been proven that the use of these starter cultures in the technology of raw ba-lik products makes it possible to obtain finished products with improved quality indicators, which is promising in the food industry. It had been shown that in the pork meat samples with the starter cultures, the acidity from 5.74 pH units is more intensively decreased – to 5.52 pH units compared to the control sample (up to 5.64 pH units). Using the starter cultures based on nitrite-reducing microorganisms for the salting in 72 hours positively influenced the formation of the required colour characteristics. It had been established that the moisture-binding capacity and plasticity of the pork meat samples with the starter cultures are characterised by the increased indicators compared with the control sample – by 4.73% and 7.73% and by 2.19 cm2.g, respectively. The difference in the volatile fatty acids content in the pork meat samples with the starter cultures compared with the control sample is 22 and 33%, respectively, in 72 hours of salting. The obtained results can be used in the enterprises of the meat processing industry to produce fermented meat products, particularly raw dried logs.

Staphylococcus caprae (S. caprae) is a gram positive, coagulase-negative Staphylococci (CoNS) that occurs as a commensal pathogen on the human skin. It recently has been recognized in causing nosocomial infections involving the bloodstream, urinary tract, heart, bone, and joints, particularly in immunosuppressed patients or individuals with prosthetic devices. Previously, S. caprae was underreported as it was difficult to identify in the clinical microbiology laboratory; however, due to advances in molecular identification methods and matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS), more clinical cases are being identified in human isolates and appropriately treated. S. caprae osteoarticular infections are usually associated with polymicrobial infections and presence of orthopedic prostheses in immunocompromised adults. This pathogen has an even rarer presentation of bone and joint infections (BJIs) in immunocompetent individuals without orthopedic devices.Our case is of a 65-year-old immunocompetent male with diet-controlled diabetes mellitus type 2 and end-stage renal disease (ESRD) on hemodialysis who presented with worsening mid-thoracic pain after a ground-level fall and was diagnosed with biopsy-proven S. caprae thoracic discitis/osteomyelitis, associated with recurrent catheter-related bloodstream infection (CRBSI). It illustrates the importance of recognizing S. caprae as an emerging human pathogen, even in immunocompetent individuals without orthopedic hardware, requiring prompt targeted treatment of native BJIs to prevent unfavorable outcomes.

Nitrates and nitrites, which are synthetic additives, are traditionally used as curing agents in meat-based products. These synthetic additives are employed in the preparation of fermented meat foods to improve quality characteristics and microbiological safety, develop distinct flavours and red-colour stability, and counteract lipid oxidation. Nitrites also display significant bacteriostatic and bactericidal action against spoilage microorganisms and foodborne pathogens (such as Clostridium botulinum and Listeria monocytogenes). However, meat curing is currently under scrutiny because of its links to cardiovascular diseases and colorectal cancer. Based on the current literature, this review provides recent scientific evidence on the potential utilisation of coagulase-negative staphylococci (CNS) as nitrate and nitrite substitutes in meat-based foods. Indeed, CNS are reported to reproduce the characteristic red pigmentation and maintain the typical high-quality traits of cured-meats, thanks to their arginine degradation pathway, thus providing the nitrite-related desirable attributes in cured meat. The alternative strategy, still based on the NOS pathway, consisting of supplementing meat with arginine to release nitric oxide (NO) and obtain a meat characterised by the desired pinkish-red colour, is also reviewed. Exploiting NOS-positive CNS strains seems particularly challenging because of CNS technological adaptation and the oxygen dependency of the NOS reaction; however, this exploitation could represent a turning point in replacing nitrates and nitrites in meat foods.

The objective of this review is to investigate the distribution and the characteristics of coagulase-negative Staphylococci (CoNS) implicated in ovine mastitis, and especially in subclinical cases, in order to provide a global perspective of the current research data and analyze specific critical aspects of the issue. PRISMA guidelines were implemented in the search of the last 20 years of the related literature in two databases. In total, 139 studies were included in this review. Relevant data were tracked down, assembled, and compared. Regarding the geographical distribution, most studies originated from Europe (68), followed by South America (33). Lacaune was the most examined breed, while S. epidermidis was the predominantly identified species, representing approximately 39% of the obtained isolates. Antibiotic resistance in the relevant bacteria was documented mostly for Penicillin (32.8%) and Amoxicillin (32.1%), while biofilm- and toxin-associated genes were encountered in variable rates because significant inequalities were observed between different articles. Significantly higher rates of antimicrobial resistance were detected in Asia and South America compared to Europe. Finally, the diagnostic procedures carried out in the respective studies were evaluated. Conventional culture and biochemical tests were mostly performed for simple strain identification; therefore, further molecular investigation of isolates should be pursued in future studies, as this will provide important data regarding specific aspects of the implication of CoNS in ovine mastitis. [ABSTRACT FROM AUTHOR]

Background: Sepsis in low-birth-weight neonates remains one of the most significant causes of neonatal morbidity and mortality. Approximately 3 million newborns suffer from sepsis globally every year. The aim of this study was to compare demographic and clinical features, as well as etiology and antibiotic susceptibility, of the main pathogens related to neonatal sepsis in two neonatal intensive units during a two-year period. Methods: We observed early-onset (EO-BSI) and late-onset bloodstream infections (LO-BSI) cases in two high-reference neonatal intensive care units (NICU) over a 24-month period (2016–2017). Samples of patients' blood were tested for the presence of the microorganisms. All bacterial isolates were tested for susceptibility to antibiotics. Results: The majority of sepsis cases weighed above 1000 g and were born by cesarean section. About 10% of the EO-BSI group died. There were differences in the EO-BSI /LO-BSI ratio in the compared wards due to differences among the admitted children. The most common pathogens isolated from blood were coagulase-negative staphylococci (CoNS) were represented by two dominating species: S. epidermidis and S. haemolyticus, followed by Klebsiella spp. strains and E.coli, which were mostly found in EO-BSI cases. No single S. agalactiae (GBS) strain was isolated. The majority of CoNS strains were resistant to methicillin, half were resistant to aminoglycosides, and one-third were resistant to macrolides and lincosamides. Half of the Gram-negative rods were resistant to beta-lactams. Conclusions: The epidemiology of sepsis in two observed NICUs is comparable to data obtained from other studies with a predominance of methicillin-resistant CoNS in LO-BSI and beta-lactam resistant E. coli in EO-BSI. It is of importance that the campaign for controlling GBS carriage in pregnant women in Poland resulted in the disappearance of GBS as a cause of sepsis. Unfortunately, there are no such measures to control E.coli related sepsis. [ABSTRACT FROM AUTHOR]

Staphylococci are the most typical bacteria found in cattle with mastitis, either Staphylococcus aureus or coagulase- negative Staphylococci (CNS). The study's goal was to determine the prevalence of Staphylococci in bovine mastitis, the antimicrobial profile, and evaluate the concentration of pro-inflammatory interleukins (IL- 4, IL-6, IL-10) related to the inflammatory response in clinical mastitis by ELISA. S. aureus (54%) and CNS (19%) were detected in 400 samples of milk from both clinical and subclinical bovine mastitis. The S. aureus isolates revealed higher resistance to ampicillin (100%), followed by amoxicillin-clavulanic acid (98.1%), cefotaxime (88.9%), erythromycin (63.2%), cefuroxime (63%), and tetracycline (61.1%). Furthermore, the CNS isolates showed high resistance against amoxicillin-clavulanic acid (100%), followed by ampicillin (94.7%), cefotaxime (89.5%), erythromycin (79.6%), cefuroxime (89.5%), and tetracycline (73.7%). However, the high susceptibility of S. aureus and CNS was observed to imipenem and ciprofloxacin. Furthermore, Cows with clinical mastitis reported high levels of IL-6 in both their serum and their milk. While they have much lower levels of IL-4 and IL-10 than normal ones (P < 0.001). In conclusion, it is recommended that laboratory results be carefully interpreted to avoid antimicrobial therapy for Staphylococci that is not clinically relevant and to ensure the advisable use of antimicrobials. Also, further study on the application of interleukins as therapeutic agents against bovine mastitis should be considered. [ABSTRACT FROM AUTHOR]

The study investigates the prevalence of Staphylococcus aureus and coagulasenegative staphylococci in raw milk and dairy products and assesses their toxin-related pathogenic potential and methicillin resistance. A total of 1015, raw milk (260) and dairy samples (325 cheeses, 180 yogurts, 140 ice creams, 110 butter samples) were collected and analyzed. The prevalence of Staphylococcus aureus and coagulase-negative staphylococci were 3.2% and 5.3% with mean counts of 3.46 and 3.16 log CFU/mL-g, respectively. Three (sea, seb, see) of five (sea, seb, sec, sed, see) staphylococcal enterotoxin (SE) genes, two (tss, etb) of four (tss, pvl, eta, etb) virulence-associated genes, and the absence of methicillin resistance (mecA) gene were defined by polymerase chain reaction. SE sea (6.9%), seb (2.3%) and see (1.1%) genes were detected in Staphylococcus aureus from one milk and seven different cheese samples. The presence of multiple enterotoxin genes (sea and see) was detected in a Staphylococcus aureus isolate from one cheese. However, the milk-sourced one coagulase-negative staphylococci possessed both the tss and etb virulence genes. The finding in this study indicates that the frequency of coagulase-negative staphylococci was higher than Staphylococcus aureus and moreover, toxin genes associated with human infections were assigned in coagulase-negative staphylococci while enterotoxin genes were determined among Staphylococcus aureus. In terms of food safety perspective, coagulasenegative staphylococci are ignored and they are not considered in standard food surveillance analysis. But the presence of virulent coagulase-negative staphylococci in foods is a public health concern. The results obtained from this study are significant as it demonstrates that pathogenic coagulase-negative staphylococci are found in foods, and provides data from Turkey. Additional research is required concerning coagulase-negative staphylococci in the food matrix and clinical isolates. [ABSTRACT FROM AUTHOR]

The ecology and diversity of resistome in coagulase-negative staphylococci (CoNS) from healthy pigs and pig farmers are rarely available as most studies focused on the livestock-associated methicillin-resistant S. aureus. This study aims to characterize the antimicrobial resistance (AMR) mechanisms, intra-host species diversity (more than one species in a host), and intra-species AMR diversity (same species with more than one AMR profile) in CoNS recovered from the nasal cavities of healthy pigs and pig farmers. One-hundred-and-one CoNS strains previously recovered from 40 pigs and 10 pig farmers from four Spanish pig farms were tested to determine their AMR profiles. Non-repetitive strains were selected (n = 75) and their AMR genes, SCCmec types, and genetic lineages were analyzed by PCR/sequencing. Of the non-repetitive strains, 92% showed a multidrug resistance (MDR) phenotype, and 52% were mecA-positive, which were associated with SCCmec types V (46.2%), IVb (20.5%), and IVc (5.1%). A total of 28% of the pigs and pig farmers had intra-host species diversity, while 26% had intra-species AMR diversity. High repertoires of AMR genes were detected, including unusual ones such as tetO, ermT, erm43, and cfr. Most important was the detection of cfr (in S. saprophyticus and S. epidermidis-ST16) in pigs and pig farmers; whereas MDR-S. borealis strains were identified in pig farmers. Pig-to-pig transmission of CoNS with similar AMR genes and SCCmec types was detected in 42.5% of pigs. The high level of multidrug, within-host, and intra-species resistome diversity in the nasal CoNS highlights their ability to be AMR gene reservoirs in healthy pigs and pig farmers. The detection of MDR-S. borealis and linezolid-resistant strains underscore the need for comprehensive and continuous surveillance of MDR-CoNS at the pig farm level. [ABSTRACT FROM AUTHOR]

Coagulase-negative Staphylococcus (CoNS) has emerged as a significant primary pathogen, frequently determined in hospitalized individuals with immune deficiencies, and regularly with indwelling or implantable clinical devices. Most infections are nosocomial, and conservative infections can lead to a variety of chronic diseases, such as urinary tract infections (S. saprophyticus), infections brought on by medical devices, endocarditis, mediastinitis, meningitis, and bloodstream infections. Although many new species have been revealed in recent years, there is still a defect in clinical details for most of that genus. Additionally, interspecies variations that must be considered make certain species more pathogenic than others. Blood cultures and other samples are commonly contaminated because CoNS has the ability to colonize human skin. Determining cases where CoNS are causal agents rather than contaminants is hence the key diagnostic challenge. However, neither phenotypic nor genetic approaches have been successful in offering a satisfactory answer to this issue. MALDI-TOF MS (99.3% of strains properly identified) was the most effective method for speedy and precise CoNS differentiation. Vitek2 paired with partial tuf gene sequencing might be an alternative that yields 100% accurate strain identification when both techniques are used together. All of the staphylococcal subspecies under investigation, include S.capitis subspecies capitis as well as subspecies urealyticus, S.cohnii subspecies urealyticus, S.saprophyticus subspecies saprophyticus, and Staphylococcus hominis subspecies novobiosepticus and subspecies. As a result, staphylococci can be routinely identified by MALDI-TOF MS without incurring significant consumable expenses or requiring time-consuming DNA sequence analysis. The revolutionary high-quality RIDOM is superior to phenotypic techniques and the NCBI database. Staphylococci are easily identified using sequencing databases, even seldom isolated species and phenotypic differences. Another challenge facing CoNS is their comprehensive antimicrobial resistance profile is relevant to clinical practice, notably in medical facilities. Consequently, genuine infections brought on by CoNS require the usage of second-line antimicrobial medications the majority of the time.

No proprietary starter cultures for crafting Chinese bacon. This study aimed to isolate Coagulase-negative Staphylococci (CNS) from Chinese bacon, identify their species, and evaluate their ability to produce biogenic amines (BAs), peptides, free amino acids (FAAs), and degrade proteins. Twenty-one isolates were deficient in hemolysis, DNase, and coagulase activities, and exhibited low amino acid decarboxylase activity. Further characterization revealed 11 CNS species showing protease, lipase, or nitrate reductase activities. Specifically, S. cohnii WX-M8 was able to degrade both sarcoplasmic and myofibrillar proteins, while S. saprophyticus MY-A10 was found to only degrade myofibrillar proteins. Both were able to reduce the BAs and increase the content of peptides around day 3. The meat fermented by these two CNS contained FAAs that are more conducive to taste formation, such as Glu and Asp, and reduced the content of bitter FAAs. These findings will provide insights into the use of CNS for Chinese bacon.

ABSTRACT Novel species of coagulase-negative staphylococci, which could serve as reservoirs of virulence and antimicrobial resistance factors for opportunistic pathogens from the genus Staphylococcus, are recognized in human and animal specimens due to advances in diagnostic techniques. Here, we used whole-genome sequencing, extensive biotyping, MALDI-TOF mass spectrometry, and chemotaxonomy to characterize five coagulase-negative strains from the Staphylococcus haemolyticus phylogenetic clade obtained from human ear swabs, wounds, and bile. Based on the results of polyphasic taxonomy, we propose the species Staphylococcus brunensis sp. nov. (type strain NRL/St 16/872T = CCM 9024T = LMG 31872T = DSM 111349T). The genomic analysis revealed numerous variable genomic elements, including staphylococcal cassette chromosome (SCC), prophages, plasmids, and a unique 18.8 kb-long genomic island SbCI ccrDE integrated into the ribosomal protein L7 serine acetyltransferase gene rimL. SbCI ccrDE has a cassette chromosome recombinase (ccr) gene complex with a typical structure found in SCCs. Based on nucleotide and amino acid identity to other known ccr genes and the distinct integration site that differs from the canonical methyltransferase gene rlmH exploited by SCCs, we classified the ccr genes as novel variants, ccrDE. The comparative genomic analysis of SbCI ccrDE with related islands shows that they can accumulate virulence and antimicrobial resistance factors creating novel resistance elements, which reflects the evolution of SCC. The spread of these resistance islands into established pathogens such as Staphylococcus aureus would pose a great threat to the healthcare system. IMPORTANCE The coagulase-negative staphylococci are important opportunistic human pathogens, which cause bloodstream and foreign body infections, mainly in immunocompromised patients. The mobile elements, primarily the staphylococcal cassette chromosome mec, which confers resistance to methicillin, are the key to the successful dissemination of staphylococci into healthcare and community settings. Here, we present a novel species of the Staphylococcus genus isolated from human clinical material. The detailed analysis of its genome revealed a previously undescribed genomic island, which is closely related to the staphylococcal cassette chromosome and has the potential to accumulate and spread virulence and resistance determinants. The island harbors a set of conserved genes required for its mobilization, which we recognized as novel cassette chromosome recombinase genes ccrDE. Similar islands were revealed not only in the genomes of coagulase-negative staphylococci but also in S. aureus. The comparative genomic study contributes substantially to the understanding of the evolution and pathogenesis of staphylococci.

Background: Peritonitis is the most important complication of peritoneal dialysis (PD) and coagulase-negative staphylococci (CNS) are a frequent cause of dialysis-related infections. The association between SCCmec typing with psm-mec positivity in staphylococci and PD-related infections has not been identified. We aim to investigate the molecular epidemiology of CNS isolated from PD-peritonitis in a single Chinese center, focusing on the genetic determinants conferring methicillin resistance. Methods: We collected 10 genetically unrelated CNS isolates from 10 patients with CNS PD-related peritonitis. The patients were divided into two groups based on the results of MIC to oxacillin: the methicillin-resistant CNS (MRCNS) and methicillin-sensitive CNS (MSCNS) groups. The biofilm formation group (BFG) and the non-biofilm formation group (NBFG) were used as the control groups. Phenotypic and molecular methods were used to analyze SCCmec types I, II and III, associated genes and biofilm formation and the existence of psm-mec. The demographic data and clinical indicators were collected. Results: Ten CNS PD-related peritonitis patients were enrolled for this study. There were 6 MRCNS and 4 MRCNS isolates. SCCmec types were fully determined in 10 isolates. Seven staphylococci (70%) carried SCCmec, of which 4 isolates carried single SCCmec type I (40%) and 3 isolates had multiple SCCmec elements (I + III). Of the 6 MRCNS isolates, 3 carried SCCmec type I (50%) and 2 isolates carried SCCmec type I + III (33.3%). A high diversity of ccr types, mec complexes and ccr-mec complex combinations was identified among the 10 CNS isolates. The psm-mec gene was detected in 2/10 (20%) CNS isolates. There was no mutation in the psm-mec gene. Conclusions: The majority of isolates were hospital-associated isolates. Furthermore, 2 psm-mec positive isolates were MRCNS in the NBFG. The PD patients frequent exposure to hospital would be the main risk factor. The presence of the psm-mec signal in the spectra of the MRCNS tested here demonstrates the presence of certain SCCmec cassettes that convey methicillin resistance. [ABSTRACT FROM AUTHOR]

The study reveals crucial information on S. capitis, a potentially dangerous bacterium that can cause sepsis in hospitalized adult patients, including its frequency and patterns of drug resistance which could significantly contribute to the existing body of knowledge in this field. This retrospective study was conducted in the King Fahad Medical City (KFMC), Riyadh, Saudi Arabia, from June 2019 to November 2022. The comprehensive data collection and analysis provide valuable information on the prevalence of S. capitis in different infection sites, antibiotic sensitivity profiles, and the association with patient demographics. A total of 219 S. capitis isolates from hospitalized patients with sepsis tested positive. Infection was most common in venous blood (139 patients, 63.5%), followed by central blood (24 patients, 11%). Other locations included venipuncture (48 patients, 21.9%) and cerebrospinal fluid (three patients, 1.4%). The results highlight the clinical significance of S. capitis as a major contributor to sepsis, especially in male patients, and emphasize the need for appropriate antibiotic selection for effective treatment. Furthermore, this study raises concerns about the high rates of methicillin resistance observed in S. capitis isolates, emphasizing the urgency in tackling antibiotic resistance and promoting judicious antibiotic prescription practices. Moreover, the identification of a decreased susceptibility to vancomycin as a potential explanation for its reduced effectiveness in treating S. capitis infections adds a crucial aspect to the discussion of treatment options and underscores the need for alternative therapeutic strategies. [ABSTRACT FROM AUTHOR]

While modern molecular methods have decisively accelerated and improved microbiological diagnostics, phenotypic variants still pose a challenge for their detection, identification and characterization. This particularly applies if they are unstable and hard to detect, which is the case for the small-colony-variant (SCV) phenotype formed by staphylococci. On solid agar media, staphylococcal SCVs are characterized by tiny colonies with deviant colony morphology. Their reduced growth rate and fundamental metabolic changes are the result of their adaptation to an intracellular lifestyle, regularly leading to specific auxotrophies, such as for menadione, hemin or thymidine. These alterations make SCVs difficult to recognize and render physiological, biochemical and other growth-based methods such as antimicrobial susceptibility testing unreliable or unusable. Therefore, diagnostic procedures require prolonged incubation times and, if possible, confirmation by molecular methods. A special approach is needed for auxotrophy testing. However, standardized protocols for SCV diagnostics are missing. If available, SCVs and their putative parental isolates should be genotyped to determine clonality. Since their detection has significant implications for the treatment of the infection, which is usually chronic and relapsing, SCV findings should be specifically reported, commented on, and managed in close collaboration with the microbiological laboratory and the involved clinicians. [ABSTRACT FROM AUTHOR]

(1) Background: Coagulase-negative staphylococci (CoNS) are an important group of organisms that can cause bloodstream infection (BSI) and infective endocarditis (IE). The prevalence of IE in patients with BSI due to different CoNS species, however, has received limited attention; (2) Methods: A retrospective study of adults with monomicrobial CoNS BSI who had undergone echocardiography and a risk factor analysis was done to determine the most common CoNS species that cause definite IE; (3) Results: 247 patients with CoNS BSI were included in the investigation; 49 (19.8%) had definite IE, 124 (50.2%) possible IE, and 74 (30.0%) BSI only. The latter two entities were grouped in one category for further analysis. The most common species in CoNS BSI was Staphylococcus epidermidis (79.4%) and most patients (83.2%) had possible IE/BSI only. 59.1% of patients with BSI due to S. lugdunensis had definite IE. The majority of CoNS were healthcare-associated/nosocomial bacteremia. Multivariable analysis demonstrated that valve disease (p = 0.002) and a foreign cardiovascular material (p < 0.001) were risk factors associated with definite IE. Patients with S. lugdunensis BSI had an 8-fold higher risk of definite IE than did those with S. epidermidis BSI and nearly a 13-fold higher risk than did patients with BSI due to other species of CoNS (p = 0.002); (4) Conclusions: The prevalence of definite IE in patients with BSI due to different CoNS species was significant. CoNS bacteremia, particularly with S. lugdunensis, confers a significant risk of IE, particularly in patients with a valve disease or intravascular foreign body material and should not be immediately dismissed as a contaminant. [ABSTRACT FROM AUTHOR]

Background: Healthcare workers may pave the way for increased infections in hospitalized patients by coagulase-negative staphylococci (CoNS). Biofilm formation and antibiotic resistance are the major problems posed by CoNS in nosocomial infections. In this study, we determined biofilm production level and the distribution of biofilm-associated and virulence genes, including icaADBC, aap, bhp, atlE, embp, and fbe, as well as IS256, IS257, mecA, and ACME clusters (arc-A, opp-3AB) among 114 clinical (n = 57) and healthcare workers (n = 57) CoNS isolates in Kerman, Iran. Results: In this study, more than 80% (n = 96) of isolates were methicillin-resistant CoNS (MR-CoNS). Out of 114 isolates, 33% (n = 38) were strong biofilm producers. Strong biofilm formation was found to be significantly different between clinical and healthcare workers' isolates (P < 0.050). In addition, 28% (n = 32) of isolates were positive for icaADBC simultaneously, and all were strong biofilm producers. The prevalence of icaADBC, mecA, bhp, fbe, and IS256 in clinical isolates was higher than that in healthcare workers' isolates (P < 0.050). A significant relationship was observed between clinical isolates and the presence of icaADBC, mecA, bhp, and IS256. Although these elements were detected in healthcare workers' isolates, they were more frequent in clinical isolates compared to those of healthcare workers. Conclusions: The high prevalence of ACME clusters in healthcare workers' isolates and biofilm formation of these isolates partially confirms the bacterial colonization in the skin of healthcare workers. Isolating MR-CoNS from healthcare workers' skin through similar genetic elements to clinical isolates, such as icaADBC, mecA, and IS256, calls for appropriate strategies to control and prevent hospital infections. [ABSTRACT FROM AUTHOR]

Staphylococci, mainly including Staphylococcus aureus and coagulase-negative staphylococci (CNS), are one of the most common pathogens causing bovine mastitis worldwide. In this study, we investigated the antimicrobial resistance and virulence profiles of staphylococci from clinical bovine mastitis in Ningxia Hui Autonomous Region of China. Antimicrobial resistance was determined by disc diffusion combined with E-test method. Genes of antimicrobial resistance and virulence factors were determined by PCR. A total of 332 staphylococcal isolates were confirmed from 1,519 mastitic milk samples, including 172 S. aureus and 160 CNS isolates. Fifteen CNS species were identified, with S. chromogenes being the most frequent found (49.4%), followed by S. equorum (13.8%). Noticeably, 2 S. agnetis isolates were found among the CNS isolates. To our knowledge, this is the first report documenting the presence of S. agnetis from bovine mastitis in China. The S. aureus and CNS isolates showed high resistance against penicillin, followed by erythromycin and tetracycline. Multidrug resistance was found in 11.6 and 16.3% of the S. aureus and CNS isolates, respectively. Resistance to penicillin was attributed to the presence of blaZ, erythromycin resistance to ermC (alone or combined with ermB) and tetracycline resistance to tetK (alone or combined with tetM). Notably, one S. equorum isolate and one S. saprophyticus isolate were both methicillin-resistant and mecA positive. Additionally, all S. aureus isolates carried the adhesin genes fnbpA, clfA, clfB, and sdrC, and most of them contained cna and sdrE. Conversely, only a few of the CNS isolates carried clfA, cna, and fnbA. Regarding toxin genes, all S. aureus isolates harbored hlb, and most of them were hlg positive. The lukE-lukD, lukM, sec, sed, sei, sen, seo, tst, seg, seh, and sej were also detected with low frequencies. However, no toxin genes were observed in CNS isolates. This study reveals high species diversity of staphylococci from clinical bovine mastitis in Ningxia Hui Autonomous Region of China. The findings for the genetic determinants of antimicrobial resistance and virulence factor provide valuable information for control and prevention of staphylococcal bovine mastitis. [ABSTRACT FROM AUTHOR]