Candida auris in various hospitals across Kuwait and their susceptibility and molecular basis of resistance to antifungal drugs.

BACKGROUND: Candida auris, a multidrug-resistant species, has the propensity of nosocomial transmission despite normal decontamination procedures. Here, we describe the isolation of C auris from patients in various hospitals in Kuwait during 2014-2018. Susceptibility to antifungal drugs and molecular basis of resistance to fluconazole, voriconazole and micafungin were also studied. METHODS: Candida auris (n = 314) obtained from 126 patients in eight hospitals were studied. All isolates were identified by PCR amplification and/or PCR-sequencing of ribosomal DNA (rDNA). Antifungal susceptibility was determined by Etest. Molecular basis of resistance to fluconazole and micafungin was studied by PCR-sequencing of ERG11 and FKS1 genes, respectively. FINDINGS: Bloodstream (n = 58), urine (n = 124), respiratory (n = 98) and other (n = 34) specimens yielded 314 C auris isolates. The proportion of bloodstream C auris among all yeast isolates was higher (42 of 307, 13.7%) in 2018 as compared to 2014-2017 (16 of 964, 1.7%) (P = .001). More bloodstream isolates (42 of 139) were cultured in 2018 than during 2014-2017 (16 of 175) (P = .001). Resistance to amphotericin B, fluconazole, voriconazole and micafungin was detected in 27.1%, 100%, 41.1% and 1.7% isolates, respectively. Fluconazole-resistant isolates contained either Y132F or K143R mutation in ERG11. Isolates with K143R mutation were additionally resistant to voriconazole. Micafungin-resistant isolates contained S639F mutation in hot spot 1 of FKS1. CONCLUSIONS: Our study highlights spreading of C auris in major hospitals across Kuwait and its increasing role as a bloodstream pathogen in 2018. Cross-resistance to voriconazole was also seen in isolates with K143R mutation in ERG11, while micafungin-resistant isolates harboured S639F mutation in hot spot 1 of FKS1.

Cyberlindnera fabianii fungaemia outbreak in preterm neonates in Kuwait and literature review.

BACKGROUND: Cyberlindnera fabianii has rarely been reported as a human pathogen. Here, we describe an outbreak of C. fabianii fungaemia involving 10 preterm neonates during a seven-month period in Kuwait and review the published reports. METHODS: Blood cultures were processed, and yeast isolates were initially identified by ID 32 C and/or VITEK 2. Molecular identification was done by PCR sequencing of internally transcribed spacer (ITS) region and D1/D2 domains of rDNA. Fingerprinting was performed with microsatellite-based and minisatellite-based primers to examine genetic relatedness among the isolates. Antifungal susceptibility testing of the isolates was done by Etest. FINDINGS: All infected neonates were preterm, received prior antibiotics and had an intravascular catheter in place. All bloodstream isolates were initially identified as Candida utilis by ID 32 C and/or VITEK 2 and showed reduced susceptibility to triazoles. PCR sequencing of rDNA identified all isolates as Cyberlindnera fabianii. Fingerprinting studies yielded identical patterns indicating clonality. One neonate died before treatment, one died during treatment, and eight neonates survived treatment with amphotericin B with/without fluconazole or caspofungin. Source of infection remained unknown despite surveillance cultures. CONCLUSION: The outbreak highlights emergence of C. fabianii as a neonatal pathogen and reinforces importance of molecular methods in its accurate identification.

Rapid and Accurate Identification of Candida albicans and Candida dubliniensis by Real-Time PCR and Melting Curve Analysis.

OBJECTIVE: Candida albicans and Candida dubliniensis are germ tube-positive pathogenic yeast species. Accurate identification of these two species is warranted since C. albicans is a highly pathogenic species while C. dubliniensis exhibits increased adherence to buccal epithelial cells, reduced susceptibility to azoles and resistance to flucytosine. We have developed a duplex real-time PCR assay for rapid detection and differentiation between clinical C. albicans and C. dubliniensis isolates. MATERIALS AND METHODS: A duplex real-time PCR assay was developed by using two species-specific primer pairs and SYBR Green dye to differentiate C. albicans and C. dubliniensis isolates via melting curve analysis of real-time PCR amplicons. Amplification products were also analyzed by agarose gel electrophoresis to confirm real-time PCR results. RESULTS: Melting temperatures (Tm) for reference strains of C. albicans and C. dubliniensis were 86.55 and 82.75°C, respectively. No amplicon was obtained with DNA from reference strains of 8 other common Candida spp. When real-time PCR was applied on 226 clinical isolates previously identified by the Vitek 2 system and/or PCR sequencing of rDNA, Tm values for C. albicans (n = 113) and C. dubliniensis (n = 98) were 86.68 ± 0.529 and 82.616 ± 0.535°C, respectively. The results were confirmed by agarose gel electrophoresis. No amplicon was obtained from 15 isolates belonging to 9 other Candida spp. CONCLUSIONS: The real-time PCR assay described here does not require prior identification of clinical yeast isolates as C. albicans/C. dubliniensis by germ tube formation and accurately reports results within 2 h. Detection of amplicons by agarose gel electrophoresis is also suitable for resource-poor settings devoid of real-time PCR facilities.

Candida fermentati as a cause of persistent fungemia in a preterm neonate successfully treated by combination therapy with amphotericin B and caspofungin.

A case of persistent candidemia in a preterm neonate caused by Candida fermentati, identified by sequencing of the internally transcribed spacer region of ribosomal DNA (rDNA), is described. The neonate was treated for 30 days by combination therapy with amphotericin B (AmBisome) and caspofungin with a successful outcome, and no drug-related side effects were observed.

Microbiological contamination of mobile phones of clinicians in intensive care units and neonatal care units in public hospitals in Kuwait.

BACKGROUND: The objective of this study was to explore the prevalence of microbiological contamination of mobile phones that belong to clinicians in intensive care units (ICUs), pediatric intensive care units (PICUs), and neonatal care units (NCUs) in all public secondary care hospitals in Kuwait. The study also aimed to describe mobile phones disinfection practices as well as factors associated with mobile phone contamination. METHODS: This is a cross-sectional study that included all clinicians with mobile phones in ICUs, PICUs, and NCUs in all secondary care hospitals in Kuwait. Samples for culture were collected from mobile phones and transported for microbiological identification using standard laboratory methods. Self-administered questionnaire was used to gather data on mobile phones disinfection practices. RESULTS: Out of 213 mobile phones, 157 (73.7 %, 95 % CI [67.2-79.5 %]) were colonized. Coagulase-negative staphylococci followed by Micrococcus were predominantly isolated from the mobile phones; 62.9 % and 28.6 % of all mobile phones, respectively. Methicillin-resistant Staphylococcus aureus (MRSA) and Gram-negative bacteria were identified in 1.4 % and 7.0 % of the mobile phones, respectively. Sixty-eight clinicians (33.5 %) reported that they disinfected their mobile phones, with the majority disinfecting their mobile phones only when they get dirty. The only factor that was significantly associated with mobile phone contamination was whether a clinician has ever disinfected his/her mobile phone; adjusted odds ratio 2.42 (95 % CI [1.08-5.41], p-value = 0.031). CONCLUSION: The prevalence of mobile phone contamination is high in ICUs, PICUs, and NCUs in public secondary care hospitals in Kuwait. Although some of the isolated organisms can be considered non-pathogenic, various reports described their potential harm particularly among patients in ICU and NCU settings. Isolation of MRSA and Gram-negative bacteria from mobile phones of clinicians treating patients in high-risk healthcare settings is of a major concern, and calls for efforts to consider guidelines for mobile phone disinfection.

Spectrum and management of rare Candida/yeast infections in Kuwait in the Middle East.

Invasive fungal infections (IFIs) are associated with high mortality rates and mostly affect patients with compromised immunity. The incidence of IFIs is increasing worldwide with the expanding population of susceptible patients. Candida and other yeast infections represent a major component of IFIs. Rare Candida/yeast infections have also increased in recent years and pose considerable diagnostic and management challenges as they are not easily recognized by routine phenotypic characteristic-based diagnostic methods and/or by the automated yeast identification systems. Rare Candida/yeasts also exhibit reduced susceptibility to antifungal drugs making proper management of invasive infections challenging. Here, we review the diagnosis and management of 60 cases of rare Candida/yeast IFIs described so far in Kuwait, an Arabian Gulf country in the Middle East. Interestingly, majority (34 of 60, 56.7%) of these rare Candida/yeast invasive infections occurred among neonates or premature, very-low-birth-weight neonates, usually following prior bacteremia episodes. The clinical details, treatment given, and outcome were available for 28 of 34 neonates. The crude mortality rate among these neonates was 32.2% as 19 of 28 (67.8%) survived the infection and were discharged in healthy condition, likely due to accurate diagnosis and frequent use of combination therapy. Physicians treating patients with extended stay under intensive care, on mechanical ventilation, receiving broad spectrum antibiotics and with gastrointestinal surgery/complications should proactively investigate IFIs. Timely diagnosis and early antifungal treatment are essential to decrease mortality. Understanding the epidemiology and spectrum of rare Candida/yeast invasive infections in different geographical regions, their susceptibility profiles and management will help to devise novel diagnostic and treatment approaches and formulate guidelines for improved patient outcome.

Wide dissemination of GES-type carbapenemases in Acinetobacter baumannii isolates in Kuwait.

Acinetobacter baumannii is an opportunistic pathogen that is an important source of nosocomial infections. Production of extended-spectrum ß-lactamases (ESBLs) of the GES type in A. baumannii has been increasingly reported, and some of these GES-type enzymes possess some carbapenemase activity. Our aim was to analyze the resistance determinants and the clonal relationships of carbapenem-nonsusceptible A. baumannii clinical isolates recovered from hospitals in Kuwait. A total of 63 isolates were analyzed, and all were found to be positive for bla(GES)-type genes. One isolate harbored the bla(GES-14) gene encoding an ESBL with significant carbapenemase activity, whereas the other isolates harbored the bla(GES-11) ESBL gene. Thirty-three isolates coharbored the bla(OXA-23) and bla(GES-11) genes. Analyses of the genetic locations indicated that the bla(GES-11/-14) genes were plasmid located. It is noteworthy that the bla(OXA-23) and bla(GES-11) genes were colocated onto a single plasmid. Nine different pulsotypes were observed among the 63 isolates. This study showed the emergence of GES-type ESBLs in A. baumannii in Kuwait, further suggesting that the Middle East region might be a reservoir for carbapenemase-producing A. baumannii.

Coniochaeta polymorpha, a new species from endotracheal aspirate of a preterm neonate, and transfer of Lecythophora species to Coniochaeta.

A new species of Coniochaeta from endotracheal secretion of a preterm neonate, Coniochaeta polymorpha, is described. This anamorphic species is characterized by development of dark brown colonies after 1 week of incubation on culture medium, formation of abundant yeast-like cells and sclerotium-like structures producing discrete, brown, nearly globose phialidic conidiogenous cells and absence of chlamydospores. A combined sequence dataset of the ITS region, partial LSU rDNA, actin and ß-tubulin genes sufficiently resolved the unique phylogenetic status of this species. In response to recent changes in the nomenclature for pleomorphic fungi, we transfer the Lecythophora species to Coniochaeta, and propose the following new combinations: Coniochaeta canina, Coniochaeta cateniformis, Coniochaeta decumbens, Coniochaeta fasciculata, Coniochaeta hoffmannii, Coniochaeta lignicola, Coniochaeta luteorubra, Coniochaeta luteoviridis and Coniochaeta mutabilis.

Emergence of methicillin-resistant Staphylococcus aureus in the Maternity Hospital, Kuwait.

OBJECTIVE: To establish the relatedness of methicillin-resistant Staphylococcus aureus (MRSA) isolates in the Maternity Hospital, Kuwait. MATERIALS AND METHODS: A total of 22 MRSA were isolated from 20 neonates and 1 mother in the Special Care Unit, Maternity Hospital, Kuwait. They were characterized using antibiogram, pulsed-field gel electrophoresis (PFGE), SCCmec typing, spa typing and multi locus sequence typing (MLST), and were screened for genes encoding Panton Valentine leukocidin (PVL) and capsular polysaccharide types 5 and 8. RESULTS: The isolates were resistant to cadmium acetate (n = 22 or 100%), trimethoprim (n = 13 or 59.1%), gentamicin (n = 7 or 31.8%), ciprofloxacin (n = 5 or 22.7%), erythromycin and clindamycin (n = 2 or 9.1%), tetracycline (n = 2 or 9.1%) and fusidic acid (n = 2 or 9.1%). Eight isolates contained genes for PVL while 15 and 6 carried genes for types 5 and 8 capsular polysaccharide, respectively. Molecular typing distinguished 12 clones. Ten of these clones consisted of 20 isolates belonging to ST60-SCCmec-IV-t3935 (5 isolates), ST6-SCCmec-IV-t6269 (4 isolates), ST194-SCCmec-IV-t6892 (3 isolates), ST1-SCCmec-V-t2962 (2 isolates) and 1 isolate each of ST77-SCCmec-IV-t339, ST935-SCCmec-V-t1084, ST1317-SCCmec-V-t1548, ST9-SCCmec-V-t5801, ST627-SCCmec-IV-t1340 and ST2148-SCCmec-IV-t2810. CONCLUSION: The study demonstrated the emergence of MRSA including novel ST60 and ST194 clones at the Maternity Hospital in Kuwait.

Genotypes and virulence genes in group B streptococcus isolated in the maternity hospital, Kuwait.

OBJECTIVE: To characterize group B streptococcus (GBS) isolates obtained from patients at the Maternity Hospital in Kuwait for their genotypes and carriage of virulence genes. MATERIALS AND METHODS: A total of 154 GBS isolates were obtained from July 1 to October 31, 2007, from vaginal swabs (n = 95), urine (n = 46), blood (n = 4) and miscellaneous sources (n = 9). Genotypes were obtained by pulsed-field gel electrophoresis (PFGE), following digestion with SmaI or EagI restriction enzymes. PCR was used to screen for the carriage of virulence genes including: surface protein of group B streptococcus (spb1), secreted fibrinogen-binding protein (fbsB), C5a peptidase (scpB), laminin-binding protein (lmb), α- (bca) and ß-subunits of the C protein (bac), resistance to protease immunity protein (rib), and phage-associated gene (pag); regulatory protein (dltR), and toxins CAMP factor (cfb), hyaluronidase (hylB) and superoxide dismutase (sodA). RESULTS: PFGE defined 14 genotypes differentiating isolates with the same serotypes into different genetic backgrounds. All isolates contained genes for virulence factors. However, cfb (99.4%), scpB (88.3%), lmb (88.3%), bca (57.8%), sodA (55.8%) and dltR (53.9%) were the common virulence genes. In total, 144 (90.3%) of the isolates contained 3 or more virulence genes. However, while cfb, lmb and scpB occurred in all genotypes, others occurred in some but not in all genotypes. CONCLUSIONS: GBS isolates obtained at the Maternity Hospital, Kuwait, belonged to diverse genetic backgrounds with the majority carrying multiple virulence genes.

Molecular fingerprinting by multi-locus sequence typing identifies microevolution and nosocomial transmission of Candida glabrata in Kuwait.

Backgrounds: Candida glabrata is a frequently isolated non-albicans Candida species and invasive C. glabrata infections in older patients are associated with high mortality rates. Opportunistic Candida infections in critically ill patients may be either endogenous or nosocomial in origin and this distinction is critical for effective intervention strategies. This study performed multi-locus sequence typing (MLST) to study genotypic relatedness among clinical C. glabrata isolates in Kuwait. Methods: Candida glabrata isolates (n = 91) cultured from 91 patients were analyzed by MLST. Repeat isolates (n = 16) from 9 patients were also used. Antifungal susceptibility testing for fluconazole, voriconazole, caspofungin and amphotericin B (AMB) was determined by Etest. Genetic relatedness was determined by constructing phylogenetic tree and minimum spanning tree by using BioNumerics software. Results: Resistance to fluconazole, voriconazole and AMB was detected in 7, 2 and 10 C. glabrata isolates, respectively. MLST identified 28 sequence types (STs), including 12 new STs. ST46 (n = 33), ST3 (n = 8), ST7 (n = 6) and ST55 (n = 6) were prevalent in ≥4 hospitals. Repeat isolates obtained from same or different site yielded identical ST. No association of ST46 with source of isolation or resistance to antifungals was apparent. Microevolution and cross-transmission of infection was indicated in two hospitals that yielded majority (57 of 91, 67%) of C. glabrata. Conclusion: Our data suggest that C. glabrata undergoes microevolution in hospital environment and can be nosocomially transmitted to other susceptible patients. Thus, proper infection control practices during routine procedures on C. glabrata-infected patients may prevent transmission of this pathogen to other hospitalized patients.

Multidrug-Resistant and Extensively Drug-Resistant Escherichia coli in Sewage in Kuwait: Their Implications.

In Kuwait, some sewage is discharged into the sea untreated, causing a health risk. Previously, we investigated the presence of pathogenic E. coli among the 140 isolates of E. coli cultured from the raw sewage from three sites in Kuwait. The aim of the current study was to characterize the antimicrobial resistance of these isolates and the implications of resistance. Susceptibility to 15 antibiotic classes was tested. Selected genes mediating resistance to cephalosporins and carbapenems were sought. ESBL and carbapenemase production were also determined. Two virulent global clones, ST131 and ST648, were sought. A total of 136 (97.1%), 14 (10.0%), 128 (91.4%), and 2 (1.4%) isolates were cephalosporin-resistant, carbapenem-resistant, multidrug-resistant (MDR), and extensively drug-resistant (XDR), respectively. Among the cephalosporin-resistant isolates, ampC, blaTEM, blaCTX-M, blaOXA-1, and blaCMY-2 were found. Eighteen (12.9%) samples were ESBL producers. All carbapenem-resistant isolates were negative for carbapenemase genes (blaOXA-48, blaIMP, blaGES, blaVIM, blaNDM, and blaKPC), and for carbapenemase production. Resistance rates in carbapenem-resistant isolates to many other antibiotics were significantly higher than in susceptible isolates. A total of four ST131 and ST648 isolates were detected. The presence of MDR and XDR E. coli and global clones in sewage poses a threat in treating E. coli infections.

Incidence, aetiology and resistance of late-onset neonatal sepsis: a five-year prospective study.

AIM: Investigate the incidence, etiological pattern and the antimicrobial resistance of late-onset neonatal infections over a period of 5 years. METHODS: Longitudinal audit of neonatal sepsis from January 2005 to December 2009, in the main maternity hospital in Kuwait. Late-onset neonatal infection was defined as the culture of a single potentially pathogenic organism from blood or cerebrospinal fluid from an infant older than 6 days in association with clinical or laboratory findings consistent with infection. RESULTS: The overall incidence was 16.9 (95% confidence interval: 15.8-18.0) episodes per 1000 live births. The commonest pathogen was coagulase-negative Staphylococcus, 339 (35.7%), while Klebsiella was the most common gram-negative infection, 178 (18.8%). Escherichia coli, Enterococcus and Enterobacter spp were each responsible for 6% of all infections. Candida caused 104 (11.0%) infections. The general pattern of infection remained unchanged over the study period. Case fatality was 11.7% (95% confidence interval: 9.7-13.9%) and was high for Pseudomonas (18.4%) and Candida (22.1%) infections. Approximately 24 and 20% of Klebsiella infections were resistant to cefotaxime and gentamicin, respectively, while 28 and 24% of Escherichia coli infections were resistant to cefotaxime and gentamicin, respectively. CONCLUSION: The incidence of late-onset infection in Kuwait is high, resembling that in resource-poor countries. The high incidence coupled with low case fatality provides an example for settings where tertiary care is introduced without strict measures against nosocomial infections. Prevention against nosocomial infections in neonatal units has the potential to further reduce neonatal mortality in these settings.

Fatal Lodderomyces elongisporus Fungemia in a Premature, Extremely Low-Birth-Weight Neonate.

Many rare yeasts are emerging as pathogens, causing invasive infections in susceptible hosts that are associated with poor clinical outcome. Here, we describe the first and fatal case of Lodderomyces elongisporus fungemia in a premature, extremely low-birth-weight neonate after spontaneous vaginal delivery. The bloodstream isolate was identified as C. parapsilosis by the VITEK 2 yeast identification system and as L. elongisporus by PCR-sequencing of the internal transcribed spacer (ITS) region of ribosomal DNA. Antifungal susceptibility testing data for the isolate, performed by the broth microdilution-based MICRONAUT-AM assay, showed susceptibility to all nine antifungal drugs tested. Despite the initiation of treatment with liposomal amphotericin B, the patient died on the same day that the blood culture yielded yeast growth. This is the first report of L. elongisporus bloodstream infection in a neonate as the previous nine cases reported in the literature occurred in adult patients. The crude mortality rate for invasive L. elongisporus infection is 50%, as only 5 of 10 patients survived.

Virulence and phylogenetic groups of Escherichia coli cultured from raw sewage in Kuwait.

In Kuwait, some untreated sewage is discharged into the sea which poses health risks. Therefore, we determined the virulence traits and the phylogenetic groups of E. coli cultured from raw sewage. Sewage was collected once every month for 12 months with culturing of a total of 140 E. coli isolates. E. coli was typed by the methods of Clermont. The five pathotypes of diarrheagenic E. coli (DEC), and extra-intestinal pathogenic E. coli (ExPEC) were detected by specific PCR assays. Four virulence genes which correlate with pathogenicity in animal models, were used for the first time for detection of ExPEC-vat (vacuolating auto-transporter toxin), fyuA (yersiniabactin receptor), chuA (heme-binding protein), and yfcV (major subunit of putative chaperon-usher fimbria). Most E. coli belonged to phylogenetic groups A (65[45%]) and B1 (34[24.3%]). Three (2.1%) isolates were DEC, while 14 (10%) isolates were ExPEC mostly in group B2 (57.1%). A relatively high prevalence of ExPEC in sewage has public health implications.

Prevalence of Chlamydia trachomatis and Neisseria gonorrhoeae among asymptomatic women attending the Capital Health region clinics in Kuwait.

BACKGROUND: Data on the prevalence of Chlamydia trachomatis (CT) and Neisseria gonorrhoeae (NG) infections are not available in Kuwait. This study was conducted to determine the prevalence of CT and NG in asymptomatic women attending 4 primary health care clinics in the Capital Health region. METHODS: A total of 9239 married women who attended the primary health care centers were offered screening tests for CT and NG over a period of 5 years. Of these, 8539 (92.4%) accepted participation. Low-vaginal secretions were collected and investigated using a molecular strand displacement amplification method. Biodata of all participating women were carefully recorded including age, parity, and nationality. Unmarried women were excluded since nonmarital sex in Kuwait is illegal. RESULTS: Of the 8539 women screened, 69.5% were Kuwaitis while 30.5% were non-Kuwaitis. About 51.3% were aged ≤40 years. The overall prevalence of CT and NG was 2.1% and 1.5%, respectively. The prevalence rates of CT and NG in Kuwaitis versus non-Kuwaitis were 1.9% and 2.3% and 1.4% and 1.6%, respectively. Both CT and NG were detected more in younger than older women; 1.6% versus 0.5% (≤40 years) and 1.2% versus 0.3% (>40 years), respectively. Higher prevalence of both infections was recorded in women in monogamous than polygamous marriages and in illiterate than educated women. CONCLUSIONS: The prevalence of CT and NG among asymptomatic women in Kuwait is low compared with rates from other Gulf Cooperation Council countries. Consequently, screening of asymptomatic women is not cost effective and is questionable.

Kodamaea ohmeri as an emerging pathogen: a case report and review of the literature.

Kodamaea (Pichia) ohmeri is a yeast-like fungus that has recently emerged as an important etiologic agent of fungemia in immunocompromised patients. We report such a case in a premature neonate born at 29 weeks of gestation. Prior to developing fungemia, she had two episodes of bacterial sepsis on day 13 and day 32 due to Enterobacter cloacae and Staphylococcus epidermidis, respectively. Kodamaea ohmeri was repeatedly isolated from blood cultures and its identity was determined by phenotypic characteristics and sequencing of the ITS and D1/D2 regions of rDNA. The neonate was successfully treated with amphotericin B. The published cases of K. ohmeri fungemia reported in pediatric patients are reviewed highlighting its increasing importance as a bloodstream pathogen.

Frequency of virulence-associated genes in Enterococccus faecalis isolated in Kuwait hospitals.

OBJECTIVE: The objective of this study was to investigate the carriage of 6 virulence-associated genes in Enterococcus faecalis isolates obtained from patients in 8 hospitals in Kuwait. MATERIALS AND METHODS: In total, 466 E. faecalis isolates were obtained from 313 urine samples, 68 wound swabs, 36 blood samples, 25 rectal swabs, 12 high vaginal swabs and 12 miscellaneous sources. Genes for gelatinase(gelE),aggregation substance (aggA), hemolysin activation factor (cylA), enhanced expression of pheromone (eep), enterococcal surface protein (esp), and E. faecalis endocarditis antigen A (efaA) were detected in PCR assays. RESULTS: Of 466 isolates, 423 (90.8%) were positive for 1 and up to 5 genes. However, none of the genes was detected in all of the isolates. The prevalence of the individual genes was eep: 31.9%; esp: 31.5%; gelE: 28.5%; efaA: 27.9%; aggA: 23.4%, and cylA: 18.5%. Of the 423 positive isolates, 148 (34.9%) were positive for 2 genes and 52 (12.3%), 15 (3.5%) and 5 (0.9%) isolates were positive for 3, 4 and 5 virulence genes, respectively. The efaA and esp combination was detected in isolates from all clinical sources. CONCLUSION: The study showed a high prevalence of virulence genes in E. faecalis isolated in Kuwait hospitals. The absence of a dominant gene in all of the isolates suggests that infections by E. faecalis may require the involvement of multiple virulence factors.

Preponderance of bla KPC-Carrying Carbapenem-Resistant Enterobacterales Among Fecal Isolates From Community Food Handlers in Kuwait.

Carbapenem-resistant Enterobacterales (CRE) are pathogens that have been found in several countries, with a significant public health concern. Characterizing the mode of resistance and determining the prevailing clones are vital to the epidemiology of CRE in our community. This study was conducted to characterize the molecular mode of resistance and to determine the clonality of the CRE fecal isolates among community food handlers (FHs) vs. infected control patients (ICPs) in Kuwait. Fecal CRE isolates obtained from FHs and ICPs from September 2016 to September 2018 were analyzed for their resistance genes. Gene characterization was carried out by polymerase chain reaction (PCR) assays and sequencing. Clonality of isolates was established by multilocus sequence typing (MLST). Of the 681 and 95 isolates of the family Enterobacterales isolated from FHs and ICPs, 425 (62.4%) and 16 (16.8%) were Escherichia coli, and 18 (2.6%) and 69 (72.6%) were Klebsiella pneumoniae, respectively. A total of 36 isolates were CRE with a prevalence of 5.3% among FH isolates and 87 (91.6%) among the ICPs. Of these, carbapenemase genes were detected in 22 (61.1%) and 65 (74.7%) isolates, respectively (p < 0.05). The detected specific genes among FHs and ICPs were positive for bla KPC 19 (86.4%) and 35 (40.2%), and bla OXA 10 (45.5%) and 59 (67.8%), in addition to bla NDM 2 (9.1%) and 32 (36.8%), respectively. MLST assays of the E. coli and K. pneumoniae isolates revealed considerable genetic diversity and polyclonality as well as demonstrated multiple known ST types and eight novel sequence types. The study revealed a relatively high number of CRE harboring predominantly bla KPC-mediated CRE among the community FH isolates vs. predominant bla OXA genes among the ICPs. Those heterogeneous CRE isolates raise concerns and mandate more efforts toward molecular surveillance. A multinational study is recommended to monitor the spread of genes mediating CRE in the community of Arabian Peninsula countries.

Monitoring antibiotic resistance profiles of faecal isolates of Enterobacteriaceae and the prevalence of carbapenem-resistant isolates among food handlers in Kuwait.

OBJECTIVES: Carbapenem-resistant Enterobacteriaceae (CRE) have become one of the most challenging problems in infectious diseases worldwide. Unrecognised personnel such as food handlers (FHs) colonised with CRE serve as a reservoir for transmission. This study assessed the prevalence and susceptibility patterns of CRE isolates from FHs working in commercial eateries in the community (CFHs) and healthcare settings (HCFHs) in Kuwait over the period 2016-2018. METHODS: Representative colonies from faecal samples were identified by API 20E and a VITEK®2 ID System. Susceptibility testing against 21 antibiotics was performed by Etest and agar dilution. RESULTS: A total of 681 isolates of the family Enterobacteriaceae were isolated from 405 FHs, of which 425 (62.4%) were Escherichia coli and 126 (18.5%) were Klebsiella pneumoniae. The prevalence of CRE among FHs was 7.7% (31/405), comprising 32% CFHs (10/31) and 68% HCFHs (21/31). Ampicillin, tetracycline and cefalotin showed very poor activities against most isolates with resistance rates of 63.3%, 41.7% and 40.8%, respectively. The prevalence of multidrug-resistant (MDR) isolates was 30.5%, including 130 E. coli (30.6%) and 22 K. pneumoniae (17.5%). An alarming level of colistin resistance (11.3%) was noted. A significant proportion of FH isolates (13.2%) exhibited extended-spectrum ß-lactamases (ESBL) phenotypes, including 80 E. coli (18.8%) and 5 K. pneumoniae (4.0%). CONCLUSION: This study revealed that asymptomatic intestinal carriage of CRE, including MDR and ESBL isolates, was relatively common in our community. It is conceivable that FHs may pose a significant risk to consumers for the acquisition and spread of resistant strains.