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Clostridioides difficile can cause colitis and is associated with hospital acquired infections. The C. difficile infection (CDI) is due to production of toxins A and B which bind to epithelial cell surface receptors and triggers signaling pathways, leading to loss of epithelial barrier function, apoptosis, and inflammation, culminating in diarrheal disease. In early days, laboratory diagnosis of CDI was based on cell culture, identification of toxins, and their cytopathic effects. These assays were replaced by enzyme immunoassays for the detection of C. difficile toxins and the GDH house-keeping gene for improved specificity. Later, molecular assays with higher sensitivity were introduced which are becoming easier to incorporate into the test algorithm. The diagnosis of CDI and significance of laboratory results can be challenging with asymptomatic colonization of C. difficile in some patients. Test result interpretation is even more challenging due to multiple guidelines, emerging resistant C. difficile ribotypes, as well as differences in disease prevalence. An accurate test result for diagnosis of CDI depends on selecting patients with high pre-test probability, collecting an acceptable stool specimen, and a thorough understanding of current test methods.

Background Antimicrobial resistance (AMR) of non-fermenting Gram-negative bacteria (NFGNB) is increasingly recognized as urgent healthcare threat. Trend data on AMR of NFGNB in Saudi Arabia are either old or limited. The objective was to estimate the prevalence and resistance trends of isolated NFGNB in Saudi Arabia. Methods A retrospective multicenter study involving seven tertiary care hospitals in Saudi Arabia was conducted between 2011 and 2016. Susceptibility testing for non-duplicate isolates was performed according to the Clinical and Laboratory Standards Institute (CLSI) guidelines in College of American Pathologists accredited diagnostic microbiology laboratories in the participating hospitals. Results Out of 461,274 isolates, 100,132 (21.7%) were NFGNB which represented 30% of gram-negative pathogens. Pseudomonas aeruginosa was the most common (73.6%), followed by Acinetobacter baumannii (21.0%) and Stenotrophomonas maltophilia (5.3%). Resistance trends of P. aeruginosa were increasing for aztreonam (absolute increase during the study was 17.3%), imipenem (12.3%), and meropenem (11.6%). A. baumannii was fully resistant to several beta lactam drugs, and resistance trends were increasing for potential treatments such as tigecycline (25.1%) and tobramycin (15.5%). S. maltophilia was >90% resistant to trimethoprim/ sulfamethoxazole and ciprofloxacin by the end of the study. Conclusion We are reporting high and/or increasing resistance of NFGNB to common treatment options. The current findings call for urgent actions to combat the increasing resistance of NFGNB. Large scale sharing of AMR data collected at different hospitals with the Saudi AMR committee would be critical to set priorities and monitor progress.

Background The Kingdom of Saudi Arabia (KSA) was the fourth country in the world to authorize the BNT162b2 coronavirus disease 2019 (COVID-19) vaccine, which it rolled out on December 17, 2020 and first targeted at healthcare workers (HCWs). This study assesses vaccine uptake among this group during the first month of its availability. Methods A national cross-sectional, pilot-validated, self-administered survey was conducted among HCWs in the KSA between December 27, 2020 and January 3, 2021. The survey included sociodemographic details, previous contact with COVID-19 patients, previous infection with COVID-19, receiving (or registering with the Ministry of Health website to receive) the COVID-19 vaccine, sources of HCWs’ information on vaccines, awareness of emerging variants of concern, and anxiety level using the 7-item Generalized Anxiety Disorder assessment. A descriptive bivariate analysis and multivariate logistic binary regression analysis were performed. The primary evaluated outcome was vaccine uptake. Results Of the 1058 participants who completed the survey, 704 (66.5%) were female, and 626 (59.2%) were nurses. Of all the respondents, 352 (33.27%) were enrolled to receive or had already received the vaccine, while 706 (66.73%) had not enrolled. In a bivariate analysis, not enrolling for vaccination was more likely in females than males (78.5% vs. 21.5%, P 40 years (70.4% vs. 29.6%, P = 0.005), Saudi HCWs than expatriates (78% vs 22%, P

Human orthopneumovirus (HOPV) is the major viral pathogen responsible for lower respiratory tract infections (LRTIs) in infants and young children in Riyadh, Saudi Arabia. Yet, predominant HOPV subtypes circulating in this region and their molecular and epidemiological characteristics are not fully ascertained. A total of 300 clinical samples involving nasopharyngeal aspirates (NPAs), throat swabs, and sputum were collected during winter seasons of 2019/2020 and 2021/2022 for HOPV subtyping and genotyping. Of the 300 samples, HOPV was identified in 55 samples (18.3%) with a distinct predominance of type A viruses (81.8%) compared to type B viruses (18.2%). Importantly, the ON1 strain of HOPV-A and BA-IX strain of HOPV-B groups were found to be responsible for all the infections. Sequence analysis revealed a duplication region within 2nd HVR of G protein gene of ON1 and BA-IX strains. This nucleotide duplication exerted a profound effect on protein length and affinity towards cell receptors. Further, these modifications may aid the HOPV in immune evasion and recurrent infections. Data from this study showed that ON-1 genotype of HOPV-A and BA-IX genotype of HOPV-B were dominant in Riyadh, Saudi Arabia. Further, a duplication of sequence within 2nd HVR of G protein gene was found.

Background: Lumbar puncture (LP) remains an essential diagnostic procedure for neurological and infectious diseases. However, it remains a source of anxiety to patients and families. This research aimed to assess the impact of a newly developed educational simplified video about LP, in the parents’ native language, that is tailored to their social background and beliefs and to assess whether it can facilitate their consent for the procedure. Methods: This prospective, interventional study was conducted at the outpatient pediatric clinics at a teaching hospital. The conventional arm used verbal explanation about LP. The second method utilized a standardized video, having the same information as the conventional arm, with streaming of graphic depictions. Parents’ knowledge and perceived LP risks were measured before and after the intervention. Results: Two hundred and one parents were enrolled, with no significant differences in the socio-demographic and baseline characteristics. Both verbal and video-based counseling were found to provide a statistically significant increase in knowledge scores, and a Wilcoxon signed-rank test showed that knowledge gains for both groups were statistically significant (Verbal Explanation: W = 2693, n = 83, P

BACKGROUND: Carbapenems are the antibiotics of last-resort for the treatment of bacterial infections caused by multidrug-resistant organisms. The emergence of resistance is a critical and worrisome problem for clinicians and patients. Carbapenem-resistant Enterobacterales (CRE) are spreading globally, are associated with an increased frequency of reported outbreaks in many regions, and are becoming endemic in many others. OBJECTIVES: Determine the molecular epidemiology of CRE isolates from various regions of Saudi Arabia to identify the genes encoding resistance and their clones for a better understanding of the epidemio-logical origin and national spread. DESIGN: Multicenter, cross-sectional, laboratory-based study. SETTING: Samples were collected from 13 Ministry of Health tertiary-care hospitals from five different regions of Saudi Arabia. METHODS: Isolates were tested using the GeneXpert molecular platform to classify CRE. MAIN OUTCOME MEASURES: Prevalence of various types of CRE in Saudi Arabia. SAMPLE SIZE: 519 carbapenem-resistant isolates. RESULT: Of 519 isolates, 440 (84.7%) were positive for CRE, with Klebsiella pneumoniae (410/456, 90%) being the most commonly isolated pathogen. The distribution of the CRE-positive K pneumoniae resistance genes was as follows: OXA-48 (n=292, 71.2%), NDM-1 (n=85, 20.7%), and NDM+OXA-48 (n=33, 8%). The highest percentage of a single blaOXA-48 gene was detected in the central and eastern regions (77%), while the bla NDM -gene was the predominant type in the northern region (27%). The southern regions showed the lowest percentages for harboring both blaOXA-48 and bla NDM genes (4%), while the western region isolates showed the highest percentage of harboring both genes (14%). CONCLUSION: The results illustrate the importance of molecular characterization of CRE isolates for patient care and infection prevention and control. Larger multicenter studies are needed to critically evaluate the risk factors and trends over time to understand the dynamics of spread and effective methods of control. LIMITATIONS: Lack of phenotypic susceptibility and clinical data. CONFLICT OF INTEREST: None.

Objectives : To investigate whether the coronavirus disease-2019 pandemic has had any effects on pediatric vaccination rates at the main university hospital in Saudi Arabia. Methods : A retrospective study conducted at King Saud University Medical City, Riyadh, Saudi Arabia using electronic health records. The vaccination statuses of all children who were scheduled for vaccinations at birth and at 2, 4, 6, 9, and 12 months during March, April and May between 2017 and 2020 were included in the study with total sample of 15,870 children, and comparisons between the cohorts were performed. Results : All vaccination visits during April and May 2020 were below the lower extremes except for the birth vaccinations. In March, April, and May 2020 there were respective drops in vaccination visits of 49.93%, 71.90% and 68.48% compared with the mean numbers of vaccination visits during the same months from 2017 to 2019. In comparisons of mean numbers of visits from March 2017 to May 2019 and March to May in 2020, the respective reductions in visits for birth and 2, 4, 6, 9 and 12-month vaccinations were 16.5%, 80.5%, 74.7%, 72.9%, 80.0% and 74.1%. Conclusions : The huge impact of the coronavirus disease-2019 pandemic on childhood vaccinations will require urgent vaccination recovery plans with innovative approaches and future action plans to maintain vaccination coverage during any subsequent pandemics. Saudi Med J 2020; Vol. 41 (11): 1197-1203 doi: 10.15537/smj.2020.11.25457 How to cite this article: Alrabiaah AA, Alshaer AH, Estrella SMC, Inclan KAS, Aljammaz HA, Almoosa KM, Alshuraym NF, Temsah MA, Alsohime FM, Alsubaie SS, Bukhari EE, Somily AM, Alzamil FA. Effects of the Coronavirus disease 2019 pandemic on routine pediatric immunization coverage rates at the main University Hospital in Saudi Arabia. Saudi Med J. 2020 Nov;41(11):1197-1203. doi: 10.15537/smj.2020.11.25457.

The possibility of superficial foot mycoses to spread through contaminated mosque carpets between worshipers imposes a great health burden and is never addressed in Riyadh, SA. We aimed to assess the prevalence of fungal infections, especially human dermatophytosis acquired from mosque carpets, in five different regions of Riyadh, Saudi Arabia and establish a means of contamination control. A cross-sectional study of 100 mosques in Riyadh, Saudi Arabia, was conducted, using a multistage random cluster sampling technique. The study included mosques that had an ablution area and regularly hosted five prayers a day. Sterile swabs were used for sampling, and the samples were transported to the laboratory for culture and analysis using a micro-scan system. A total of 199 (65%) samples contained several fungal species. Rhodotorula (74%), Cladosporium (47%), Bipolaris (46.6%), other yeasts (43.7%), Alternaria (40%), dematiaceous molds (34%), Curvularia (32.4%), and Candida (31%) were the most frequently isolated species. Species belonging to several other genera were also detected. This study revealed a relatively high prevalence of fungal organisms in mosque carpets in Riyadh, suggesting the need for implementing new strategies and laws to increase the level of hygiene awareness among worshipers and mosque caretakers to limit the spread of foot fungal infections.

Klebsiella pneumoniae carbapenemase (KPC)-producing Enterobacteriaceae have been isolated from human patients in many countries across the globe but rarely in Saudi Arabia. Here we provide the genomic characterization of the first KPC-producing K. pneumoniae isolated from the urine of a patient in Riyadh, Saudi Arabia, who had a recent travel history to Egypt involving a medical procedure. Presence of KPC-encoding gene initially detected with the Xpert Carba-R assay was confirmed by traditional PCR. Susceptibility testing using the VITEK 2 system, E-test and microbroth dilution methods showed that the K. pneumoniae isolate, namely SA01_KPC-2, was resistant to all antibiotics except colistin and ceftazidime/avibactam. Whole-genome sequencing (WGS), performed on the Illumina Miseq instrument, identified the isolate as sequence type (ST) 383 and serotype KL30-D1 O1v2. Genome assembly of SA01_KPC-2 indicated the presence of two plasmids. Plasmid pSA01_KPC-2, of approximately 45.9 kb in size, harbored the blaKPC-2 flanked by ISKpn27 and ISKpn6 and had a backbone similar to published KPC-carrying IncX6 plasmids. The second plasmid pSA01_incHIB1, a derivative of published 372-kb plasmid pKpvST383, carried genes encoding virulence factors and resistance to five classes of antibiotics. The isolation of the first KPC producer in Saudi Arabia requires high attention and rapid interventions to prevent further spread. Keywords: Carbapenem-resistant enterobacteriaceae (CRE), Klebsiella pneumoniae carbapenemase (KPC), IncX6 plasmids, ST383, Saudi Arabia

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is highly contagious with various possible routes of transmission, resulting in high mortality globally. Controversy exists regarding the vertical transmission of the SARS-CoV-2 infection to fetuses of COVID-19-infected women. The aim of this study was to investigate the possibility of the vertical transmission of SARS-CoV-2 from COVID-19-infected mothers to their neonates.We prospectively collected demographical and clinical characteristics of 31 COVID-19 positive pregnant women and their neonates. All mothers and neonates were tested for SARS-CoV-2 infection using the real-time polymerase chain reaction on nasopharyngeal swabs and breast milk samples. Antenatal and placental abnormalities were ultrasonically and histopathologically examined. In cord blood samples, the immunoglobins (Ig) M and IgG were estimated qualitatively.The women's mean age and gestational age were 31 years and 38 weeks, respectively, with 58% undergoing an elective cesarean section. Gestational diabetes was reported in 29% of cases, 64.5% of women were medically free and only 16.12% were symptomatic. A normal antenatal ultrasound was observed in 77.42% of cases. Nine cord blood samples were positive for IgG. Villous infarction (24%), villous agglutination, and chorangiosis (51%), accelerated villous maturation (21%) and reduced and hypercoiling were reported for 6.97% of the umbilical cords. Three newborns had possible vertical transmission of SARS-CoV-2 infection, of which, two were preterm and IUFD. The third neonate was born full-term, admitted to NICU and later discharged in good health.Our findings support the possibility of the direct vertical transmission of the SARS-CoV-2 infection to neonates from infected mothers. Further studies with a larger sample size are required to validate the current findings.

Background Streptococcus anginosus is an emergence opportunistic pathogen that colonize the human upper respiratory tract (URT), S. anginosus alongside with S. intermedius and S. constellatus, members of S. anginosus group, are implicated in several human infections. However, our understanding this bacterium to the genotype level with determining the genes associated with pathogenicity and antimicrobial resistance (AMR) is scarce. S. anginosus 47S1 strain was isolated from sore throat infection, the whole genome was characterized and the virulence & AMR genes contributing in pathogenicity were investigated. Methodology The whole genome of 47S1 was sequenced by Illumina sequencing technology. Strain 47S1 genome was de novo assembled with different strategies and annotated via PGAP, PROKKA and RAST pipelines. Identifying the CRISPR-Cass system and prophages sequences was performed using CRISPRloci and PhiSpy tools respectively. Prediction the virulence genes were performed with the VFDB database. AMR genes were detected in silico using NCBI AMRFinderPlus pipeline and CARD database and compared with in vitro AST findings. Results β-hemolytic strain 47S1 was identified with conventional microbiology techniques and confirmed by the sequences of 16S rRNA gene. Genome of 47S1 comprised of 1981512 bp. Type I-C CRISPR-Cas system and 4 prophages were detected among the genome of 47S1. Several virulence genes were predicted, most of these genes are found in other pathogenic streptococci, mainly lmb, pavA, htrA/degP, eno, sagA, psaA and cpsI which play a significant role in colonizing, invading host tissues and evade form immune system. In silico AMR findings showed that 47S1 gnome harbors (tetA, tetB & tet32), (aac(6′)-I, aadK & aph(3′)-IVa), fusC, and PmrA genes that mediated-resistance to tetracyclines, aminoglycosides, fusidic acid, and fluoroquinolone respectively which corresponds with in vitro AST obtained results. In conclusion, WGS is a key approach to predict the virulence and AMR genes, results obtained in this study may contribute for a better understanding of the opportunistic S. anginosus pathogenicity.

INTRODUCTION Net fluid balance and its role in sepsis-related mortality is not clear; studies suggest that aggressive fluid resuscitation can help in treatment, whereas others consider it is associated with poor outcomes. This study aimed to clarify the possible association of initial 24 hours' fluid balance with poor outcomes in pediatric patients with sepsis. METHODS Retrospective data analysis included pediatric patients admitted with suspected or proven sepsis or septic shock to pediatric intensive care unit (PICU) of a tertiary care teaching hospital in Saudi Arabia. RESULTS The study included 47 patients; 13 (28%) died, and mortality rate was significant in children with neurologic failure (P < 0.02), mechanical ventilation within 24 hours of admission (P < 0.03), leukopenia (P < 0.02), abnormal international normalized ratio (P < 0.02), initial blood lactate levels higher than 5 mmol/L (P < 0.02), or positive fluid balance at 24 hours of admission to the PICU (P < 0.001). CONCLUSION Among children with sepsis and/or septic shock, there is significant association between mortality and initial high blood lactate levels and positive fluid balance at 24 hours from admission to the PICU.

Objectives: To identify pulmonary tuberculosis (PTB) delayed inpatient diagnosis duration and contributing factors in an academic center in Saudi Arabia (SA). Methods: Retrospective review of all culture-confirmed PTB cases between May 2015 and April 2019. The outcomes were the timing between admission and suspicion of PTB or isolation to either early group (within 24 hours of admission) and late group (24 hours after admission). Results: Forty-nine cases were included with a median age of 49 years; a third of them were above 65 years of age. Most patients were of Saudi nationality and male. Approximately 38% of the cases were in the delayed group, half of them were smear-positive, with an average delay of 5.5 days. This was significant with age above 65 years (odds ratio [OR]=8.93, 95% confidence interval [CI]=2.22-35.95) presence of non-respiratory symptoms (OR=5.6, 95% CI=1.56-19.98), malignancy (OR=13.38, 95% CI=1.46-122.71), chronic medical problems (OR=4.90, 95% CI=1.31-18.32), missed chest x-ray findings (OR= 48, 95% CI=8.63-266.88) or procalcitonin level above 0.5 ng/mL (OR=12, 95% CI=1.58-91.08). Conclusion: Physicians in SA need to have a low threshold for PTB consideration in elderly patients or those with a history of malignancy. A careful review of the initial chest x-ray might help to overcome missing cases of PTB.

Background: The purpose of this study was to assess the risk of venous thromboembolism (VTE) and the potential need for thromboprophylaxis in patients with tuberculosis (TB). Methods: A total of 103 patients who underwent treatment for TB at the King Khalid University Hospital in Riyadh between February 2015 to May 2018, the percentage of patients diagnosed with TB who developed VTE was assessed. This was a retrospective cohort study conducted at King Khalid University Hospital in Riyadh. Fisher's exact test was used to analyze the categorical variables.P < 0.05 was considered statistically significant. Results: Our data showed the prevalence of VTE in TB patients to be 2.93% (3/103). VTE occurred irrespective of the type of TB (pulmonary and/or extrapulmonary). All TB patients with VTE showed no significant association with factors such as human immunodeficiency virus coinfection, malignancy, and multidrug-resistant TB. Conclusion: Our study showed that TB patients in Saudi Arabia may be at a higher risk for developing VTE and should be carefully monitored as potential candidates for additional thromboprophylaxis; further studies are recommended to establish more reliable assessment and recommendations.

Introduction: Middle East respiratory syndrome coronavirus (MERS-CoV) continues to cause frequent outbreaks in hospitals in Saudi Arabia. Since healthcare workers (HCWs) have a higher risk of acquiring and spreading MERS-CoV, we aimed to evaluate the perceived risk and anxiety level of HCWs in Saudi Arabia regarding MERS. Methodology: An anonymous, self-administered questionnaire was sent online to HCWs at King Khalid University Hospital in Saudi Arabia. The total knowledge and anxiety scores were calculated. Logistic regression analyses were used to identify predictors of high anxiety scores. Results: Of 591 (70%) HCWs that responded, 284 (55%), 164 (32%), and 68 (13.2%) were physicians, nurses, and technicians, respectively. Physicians obtained a lower median knowledge score (6/9) compared to other professions (7/9). The mean anxiety score was similar for physicians and other HCWs (3/5); however, non-physicians expressed higher levels of anxiety toward the risk of transmitting MERS-CoV to their families, with an anxiety score of 4/5. The ability of the virus to cause severe disease or death was the most frequently reported reason for worry by physicians (89.7%) and non-physicians (87.2%). Overall, 80% of physicians and 90% of non-physicians reported improvement in adherence to hand hygiene and standard precautions while in hospital (p = 0.002). Concern over transmitting MERS-CoV to family members was the most predictive factor for anxiety among non-physician HCWs. Conclusion: A significant proportion of HCWs expressed anxiety about the risk of acquiring MERS-CoV infection. Healthcare institutions need to develop an integrated psychological response for HCWs to the occupational and psychological challenge of MERS-CoV outbreaks.

Purpose There is a need for continuous surveillance of methicillin-resistant Staphylococcus aureus (MRSA) to identify emergence of new strains. We hypothesize that MRSA strains are evolving with ongoing acquisition of SCCmec elements. This study was carried out to evaluate the evolution of MRSA at a tertiary care facility in Saudi Arabia. Methods MRSA isolates associated with invasive clinical infection, which were identified in 2017 at the microbiology laboratory, King Khalid University Hospital (KKUH) in Riyadh, Saudi Arabia, were studied. The molecular characterization of isolates was carried out using StaphyType DNA microarray (Alere Technologies GmbH/Abbott, Jena, Germany). Results The 125 MRSA isolates studied belonged to 18 clonal complexes (CC) which were distributed into 32 strain assignments. The predominant CC were CC5 (n=30), CC6 (n=17), CC80 (n=13), CC22 (n=12), CC361 (n=12). The findings demonstrated the first identification of CC152, CC361 and CC1153 MRSA as well as ST5-MRSA-[I+fus], "Geraldine Clone", CC6-MRSA-IV (PVL+) and CC88-MRSA-V (PVL+), WA MRSA-117 in Saudi Arabia. Four novel variants were identified: CC5-MRSA-[VI+fus+tirS], CC22-MRSA-[V/VT+fus](PVL+), CC152-MRSA-[V+fus](PVL+) and CC361-MRSA-[VT+fus]. Fifty-four isolates (n/N=54/125; 43.2%) including the novel strains carried the Q6GD50 SCCfusC gene while the Panton-Valentine leukocidin genes were present in 30.4% (n/N=38/125). Conclusion The findings demonstrate an expanding MRSA repertoire in our setting including emergence of previously unreported clonal complexes and novel strains. The high carriage of fusC gene suggests a role for fusidic acid misuse in driving the evolution of the MRSA genome and underscores the need for increased monitoring of antibiotic use.

To assess the antibacterial activity of ceftobiprole against Methicillin-resistantA retrospective cohort study with a total of 49 MRSA and 99Approximately 100% of the MRSA isolates were susceptible with MIC50/90 value of 1/1.5 mg/L while 69.8% of multi-drug resistant (MDR)The excellent activity of ceftobiprole against MRSA would have major implications in management of the patients with serious infections, as an empirical treatment or alternative to vancomycin. Ceftobiprole has a very low activity against MDR

Background: Direct or indirect hand contact plays a significant role in health-care-associated infections. Family members of pediatric patients may have various hand hygiene practices. We aimed to evaluate the perceptions of hand hygiene and handshaking practices among family caregivers of hospitalized children in a tertiary care hospital. Methods: A cross-sectional study was conducted using a pretested two-part questionnaire for a randomly selected caregiver of hospitalized children at King Khalid University Hospital, Riyadh, Saudi Arabia, between January 21 and March 8, 2018. The survey data were analyzed using SPSS version 21. Results: One hundred and eighty caregivers voluntarily participated in the study. The mean age was 35.1 years, and female parents comprised 85.6% of the sample. The majority of participants (82.8%) did not receive any previous formal education on hand hygiene. Most of the participants (relative importance index of 84.6%) had correct answers on a modified version of the World Health Organization “Perception Survey for Health-Care Workers.” However, handshaking avoidance was low in general, with caregivers reporting their handshaking practices did not change even when dealing with people who have flu-like symptoms. There was no significant correlation between the participants' characteristics and handshaking avoidance practice, except for those who were working in the medical field, who showed a significantly higher handshaking avoidance. Conclusion: Formal education for hospitalized children's caregivers on hand hygiene and handshaking practice is lacking. Information on appropriate hand hygiene and potential risks of infection spread is required, especially for those parents without a previous medical background.

Novel COVID-19 infections caused major morbidity and mortality globally in the adult age group. Likewise, SARS-COV-2 infections in children are highly risky in the selected patient population. We performed a focused literature search of published reports from December 1, 2019, till August 20, 2020. The aim was to explore the etiology, clinical presentations, and outcome of pediatric COVID-19 patients. Viral respiratory infections are associated with high societal costs for children. In addition, children with asymptomatic SARS-COV-2 infections can be a source of COVID-19 spread to parents and caregivers. The major reported risk factors for pediatric COVID-19 cases were close contact with a SARS-COV-2 positive family member, a history of travel, and/or living in endemic areas. Children with COVID-19 who required ICU care had various comorbidities, such as malignancy. As the pandemic evolved, multiple cases of multisystem inflammatory syndrome in children and adolescents temporarily related to covid-19 (MIS-C) were reported. A unique population is neonates born to COVID-19 affected mothers, as there is an urgent need to optimize their management and outcome during this rapidly evolving pandemic. The early identification of SARS-COV-2 infection in infants and children has important direct management effects in these children and public health implications because of the effects on disease transmission control measures.

Middle East respiratory syndrome coronavirus (MERS-CoV) is a member of the beta-coronavirus genus of zoonotic origin that emerged in the Arabian Peninsula and is associated with significant morbidity and mortality. This study was conducted to assess the plasma levels of cytokines to evaluate the Th1/Th2 status among 46 MERS-CoV-infected patients (19 asymptomatic and 27 symptomatic) and 52 normal healthy controls using a customized luminex kit. Comparative analysis of data between MERS-CoV-infected patients and normal healthy controls revealed that although no difference was observed between asymptomatic MERS-CoV patients and controls, the mean plasma levels of interleukin (IL)-10 (44.69 ± 40.04 pg ml−1 versus 14.84 ± 6.96 pg ml−1; P < 0.0001), IL-4 (22.46 ± 8.02 pg ml−1 versus 16.01 ± 9.97 pg ml−1; P < 0.0001), IL-5 (10.78 ± 2.86 pg ml−1 versus 8.06 ± 1.41 pg ml−1; P < 0.0001) and IL-13 (14.51 ± 3.97 pg ml−1 versus 11.53 ± 4.16 pg ml−1; P < 0.003) in MERS-CoV symptomatic patients were significantly higher than the normal controls. The mean plasma levels of interferon (IFN)-γ and IL-12 were no different among the study groups. The cytokine profile among symptomatic MERS-CoV-infected patients was skewed to a Th2 type immune response.

Introduction: The use of mobile phones by healthcare workers is a risk factor for microorganism transmission in healthcare settings. Pathogenic bacteria such as methicillin-resistant Staphylococcus aureus and gram-negative bacteria like Escherichia coli that are known to cause nosocomial infection have been isolated from mobile phones. In this cross-sectional study, we assess the burden and related risk factors of the bacterial colonization of healthcare workers’ mobile phones. Methodology: We collected samples from the mobile phones of 130 healthcare workers’ in a Saudi Arabian teaching hospital, using moistened cotton swabs. The isolated organisms were identified using an automated identification and susceptibility system. Multiple logistic regression analysis was used to test the data. Results: Of 130 swabs collected, 45 (34.6%) grew one species and 48 (36.9%) grew two or more. Staphylococcus epidermidis was the most commonly isolated bacteria (52.3%), followed by Micrococcus and related species (25.4%), Staphylococcus hominis (13.8%), and Bacillus species (6.9%). Clinically significant microorganisms such as S. aureus and Pseudomonas sp. were identified in 2 (1.5%) samples, respectively. The odds of mobile phone colonization were 8.5 times higher (95% CI = 3.2-23.1) in the laboratory, neonatal intensive care unit, and medicine departments. Mobile phones owned for more than one year were more likely to be culture positive (OR = 2.9, 95% CI = 1.1-7.6). Conclusions: In our study, the prevalence of bacterial colonization among healthcare workers’ mobile phones was high. Our findings suggest that high-risk groups for mobile phone colonization—such as laboratory, neonatal intensive care unit, and medicine department staff—should be a priority for preventative measures, to improve infection control.

Background and objectives: Middle East Respiratory Syndrome Corona Virus (MERS-CoV) outbreak in 2014 was associated with high public anxiety in the affected countries. Media speculations may have increased this psychological distress. The healthcare community was the most distressed because they were at the highest risk of infection. This study is the first to explore MERS-CoV epidemic impact on medical students’ perception and determinants of their psychological distress during this outbreak. Methods: We randomly selected and surveyed 200 students from the College of Medicine at King Saud University, Riyadh, Saudi Arabia. A predesigned questionnaire was answered by participants, and the collected data were statistically analyzed. Results: One hundred and seventy-four (87%) responded. Female students had a significantly higher mean stress level than males (P < 0.001). Participants had a mean GAD score of 2.7 ± 3.1 and a median of 2. Perceived sufficiency of information score was the highest mean and median (17.4 ± 4.2 and 18 respectively). College and hospital announcements were the most common source of information (25.4%). One hundred and thirty-four (77%) reported minimal anxiety, thirty-two (18.4%) reported mild anxiety, 8 (4.6%) reported moderate anxiety, and none of them reported severe anxiety (score >14). The stress level (as reported on 1–10 scale) shows significant correlation with Generalized Anxiety Disorder (GAD-7) score. We found in this study that significant predictors in our model, in terms of more significant to the least, were: an increased self-report on hygienic habits, self-reported social avoidance, the generalized anxiety score and finally being female gander while other variables including numbers of resources access, agreeing with public fear and knowledge score on MERS-CoV all were found to be non-significant. However, the number of accessed resources, as per students, has borderline significant correlation with higher self-reported anxiety from MERS-CoV. Conclusions: Medical students’ psychological needs during the MERS-CoV outbreak should be addressed appropriately. Our results highlight the need to establish psychological support programs for medical students during an infectious disease outbreak. Keywords: MERS-CoV, Psychological stress, Medical students, Saudi Arabia

Background The global pandemic of coronavirus disease of 2019 (COVID-19) has led to unprecedented psychological stress on health workers (HCWs). We aimed to assess the psychological impact of COVID-19 on HCWs in comparison to the stress brought on by the Middle East respiratory syndrome coronavirus (MERS-CoV) epidemic in Saudi Arabia. Method Between February 5th and 16th, 2020, 811 health-care workers (HCWs) of a tertiary care teaching hospital were invited to fill a questionnaire regarding concerns and worries about the novel coronavirus pandemic, along with Generalized Anxiety Disorder (GAD-7) Anxiety Severity screening tool. Results Out of 582 HCWs who completed the survey questionnaire (response rate of 71.8%), about 40% were exposed previously to MERS-CoV infected or suspected patients during a previous hospital outbreak. While there were no COVID-19 cases reported yet in Saudi Arabia at the time of data collection, still, the anxiety level from COVID-19 was significantly higher than that from MERS-CoV or seasonal influenza: 41.1% were more worried about COVID-19, 41.4% were similarly worried about both MERS-CoV and COVID-19, and 17.5% were more stressed by the previous MERS-CoV hospital outbreak. The most frequent concern was transmitting the infection to family and friends (2.71/5) than to themselves only (2.57/5). Conclusion Pandemic and epidemic infectious diseases such as COVID-19 or MERS-CoV impose a significant level of anxiety and stress on healthcare workers who are caring of infected patients, with their main concern being the risk of transmitting the infection to their families or to acquire it themselves. Therefore, optimizing the compliance of healthcare workers with the proper infection prevention and control measures is paramount during the infectious disease outbreak, to ensure their safety, to decrease the likelihood of getting infected or transmitting the infection to others, and consequently to alleviate their psychological stress and anxiety.

BackgroundIt has been established that the urinary tract is not sterile, however, research related to the study of urine bacteria are limited. Our work aims to study the frequency and patterns of resistance of normal urinary aerobic bacterial flora.MethodsClean catch midstream urine specimens were collected from 120 young healthy females, and then cultured. Identification of bacteria and antimicrobial susceptibility were done by means of Biomérieux VITEK® 2 automated systems. Subjects who have undergone antimicrobial treatment in one month weren’t included.ResultsThe incidence of positive bacterial cultures was 54.2%, of which 21.5% were ploymicrobial. 107 bacterial isolates that encompass 12 genera and 27 species that were predominated by Gram-positive bacteria (72%) were cultivated. Staphylococcaceae (46.1%) and Enterobacteriaceae (17.8%) were the most frequently isolates among Gram positive and Gram negative bacteria respectively, from them 36 species have been identified as b lactamase producers. The top four frequently isolated bacteria are Micrococcus species (16%), Staphylococcus haemolyticus (13.2%), Staphylococcus aureus (10%), and Klebsiella pneumoniae (10%). Twenty two bacterial species were subjected to antimicrobial susceptibility testing by using broad and narrow spectrum antibiotics and antimicrobials. Ampicillin showed the lowest susceptibility rate against Gram positives, followed by erythromycin and azithromycin. The lesser antimicrobial susceptibility potential among Gram negative bacteria was exhibited against ampicillin, followed by piperacillin and cefotaxime.ConclusionOur findings emphasize the importance of highlighting urine bacterial flora in research especially those related to susceptibility patterns, employing more advanced culture methods as multiple drug resistant bacteria were isolated.

Objectives: To evaluate the presence of Coxiella burnetii (C. brunetii) infection among patients presenting with fever of unknown origin (FUO). Methods: A cross-sectional study of 100 patients (54 men and 46 women; mean age: 34.3 ± 19.2 years) with FUO was conducted at King Khalid University Hospital, Riyadh, Saudi Arabia between March 2015 and June 2016. Phase 1 and phase 2 C. burnetii -specific antibodies in serum samples were detected by enzyme-linked immunosorbent assay. Results: Coxiella burnetii phase 1 and phase 2 antibodies were detected in 16% of the patients. Phase 2 IgM was present in 2% of the patients, whereas phase 2 IgG antibodies were detected in 11% of the patients. Coxiella burnetii -specific phase 1 IgG was found in 2% of the patients, and 8% of the patients harbored phase 1 IgA antibodies in their serum. Conclusion: The presence of C. burnetii -specific antibodies in many patients suffering from FUO highlights the importance of Q fever screening among patients presenting with febrile illness.

Introduction: Healthcare workers (HCWs) colonized with Staphylococcus aureus may serve as a reservoir of infection. This study was carried to determine the genetic make-up of S. aureus nasal colonizers in HCWs. Methodology: Nasal swabs were obtained from 93 HCWs and molecular characterization of identified S. aureus isolates was carried out using the StaphyType DNA microarray (Alere Technologies GmbH, Jena, Germany). Results: Twenty-nine HCWs (31%) were colonized with S. aureus (MSSA = 23; MRSA = 6). Thus the overall MRSA carriage rate was 6.5% (n/N = 6/93) and 20.7% (n/N = 6/29) of those colonized with S. aureus harboured MRSA. The S. aureus isolates belonged to 16 clonal complexes (CC). MSSA isolates included three each for CC15, CC188, ST2867; two each for CC5, CC97, CC367 as well as one each for CC1, CC8, CC30, CC45, CC101, CC121, ST291/813 and CC1153. The staphylococcal cassette chromosome recombinase genes ccrA-1; ccrB-1 and the fusidic acid resistance gene (fusC) were present in two MSSA isolates (CC1 and CC8). The six MRSA isolates included CC5-MRSA-[VI+fusC] (n = 2); one each of CC5-MRSA-V; CC22-MRSA-IV (tst1+); CC80-MRSA-IV [pvl+] (“European CA-MRSA Clone”) and CC97-MRSA-[V+fusC]. Conclusion: There is wide clonal diversity of S. aureus colonizers with associated high MRSA carriage among the HCWs. The presence of genetically stable MSSA isolates with the capability to transform into MRSA isolates is of concern.

BACKGROUND: Bloodstream infections are associated with high rates of morbidity and mortality. Rapid detection of bloodstream infections is important in achieving better patient outcomes. OBJECTIVE: Compare the time-to-detection (TTD) of the new BacT/Alert Virtuo and the BACTEC FX automated blood culture systems. DESIGN: Prospective simulated comparison of two instruments using seeded samples. SETTING: Medical microbiology laboratory. METHODS: Blood culture bottles were seeded in triplicate with each of the standard ATCC strains of aerobes, anaerobes and yeast. TTD was calculated as the length of time from the beginning of culture incubation to the detection of bacterial growth. MAIN OUTCOME MEASURES: TTD for the various tested organisms on the two microbial detection systems. RESULTS: The 99 bottles of seeded blood cultures incubated in each of the blood culture systems included 21 anaerobic, 39 aerobic and 39 pediatric bottles. The BacT/Alert Virtuo system exhibited significantly shorter TTD for 72.7 % of the tested organisms compared to BACTEC FX system with a median difference in mean TTD of 2.1 hours (interquartile range: 1.5-3.5 hours). The BACTEC FX system was faster in 15.2% (5/33) of microorganisms, with a median difference in mean TTD of 25.9 hours (IQR: 9.1-29.2 hours). CONCLUSION: TTD was significantly shorter for most of the microorganisms tested on the new BacT/Alert Virtuo system compared to the BACTEC FX system. LIMITATIONS: Use of simulated cultures to assess TTD may not precisely represent clinical blood cultures. CONFLICT OF INTEREST: None.

Actinomycosis is a rare infection in patients younger than 10 years of age. It mainly affects the cervicofacial region, but many other sites of infection have been recognized. About 70% of infections are due to either Actinomyces israelii or Actinomyces gerencseriae.Actinomyces neuii was first described in 1985 in two patients with post cataract endophthalmitis, A. neuii represents 17% of clinical Actinomyces isolates. Several reports indicated a well-known association between Actinomyces infections and Intrauterine devices (IUD).We are reporting a case of neonatal sepsis due to A. neuii as a first case reported from Saudi Arabia. It was thought to be the cause of the premature labor and neonatal sepsis.The prevalence of Actinomyces infection is likely underestimated and additional premature labors and abortions could have been caused by Actinomyces infections that were never detected. More studies are needed to confirm the association of maternal Actinomyces infections with preterm labor. Keywords: Actinomyces neuii, Newborn, Sepsis

Purpose Methicillin resistant Staphylococcus aureus CC15 strains (CC15-MRSA) have only been sporadically described in literature. This study was carried out to describe the genetic make-up for this rare MRSA strain. Methods Four CC15-MRSA isolates collected in Riyadh, Saudi Arabia, between 2013 and 2014 were studied. Two isolates were from clinical infection and 2 from retail meat products. Whole genome sequencing was carried out using Illumina HiSeq2500 genome analyzer. Results All the CC15-MRSA isolates had the multilocus sequence typing profile ST1535, 13-13-1-1-81-11-13, which is a single locus variant of ST15. Of the 6 contigs related to the SCC element, one comprised a recombinase gene ccrAA, ccrC-PM1, fusC and a helicase, another one included mvaS, dru, mecA and 1 had yobV and Q4LAG7. The SCC element had 5 transposase genes, namely 3 identical paralogs of tnpIS431 and 2 identical paralogs of tnpIS256. Two identical copies of a tnpIS256-based insertion element flank the aacA-aphD gene. Two copies of this insertion element were present with 1 located in the SCC element and another inserted into the sasC gene. A short 3 kb region, which lacks any bacteriophage structural genes and site-specific DNA integrase, was inserted into the hlb gene. The hsdM and the 5'-part of the hsdS gene are replaced by a copy of the hsdM/hsdS paralogs from νSaβ giving rise to a new chimeric paralog of hsdS in νSaα. Conclusion CC15-MRSA shows a novel SCCmecV/SCCfus composite element. Its variant of hsdM/hsdS probably facilitated uptake of foreign mobile genetic elements that promoted emergence of CC15-MRSA. Close surveillance is needed to monitor spread and emergence of further CC15 MRSA strains.

To quantitatively evaluate the rate, type, and level of contamination of anterior cruciate ligament (ACL) hamstring autografts after harvesting and preparation or dropping onto the operating room (OR) floor. Two hamstring autograft specimens were prospectively retrieved from each graft in a consecutive series of 50 patients undergoing primary isolated ACL reconstruction (100 specimens total). One specimen was retrieved immediately after harvesting and dropped onto the OR floor (dropped group). The other was retrieved just after graft implantation and before fixation (control group). Each specimen was incubated for aerobic and anaerobic growth, and the number of colony-forming units (CFU)/g was measured. Patients’ clinical course was monitored for signs of surgical site infection (SSI). The control and dropped groups had positive culture rates of 11/50 (22%) and 16/50 (32%), respectively, with no significant difference between groups (n.s.). The most common organism in the control group was Staphylococcus epidermidis (45.5%) followed by S. aureus (36.4%). In the dropped group, the most common organism was S. epidermidis (31.3%) followed by Bacillus species (25%). The median (range) CFU/g among positive specimens in the dropped and control groups was 65 (8–150) and 10 (2–60), respectively (P = 0.0003). No patient developed postoperative SSI. Intraoperative hamstring autograft contamination rates were high. Hence, routine prophylactic decontamination of all hamstring autografts after harvesting and preparation and before implantation is recommended. Controlled laboratory study.

Since discovery of Middle East respiratory syndrome coronavirus (MERS-CoV), a novel betacoronavirus first isolated and characterized in 2012, MERS-CoV real-time reverse transcriptase polymerase chain reaction (rRT-PCR) assays represent one of the most rapidly expanding commercial tests. However, in the absence of extensive evaluations of these assays on positive clinical material of different sources, evaluating their diagnostic effectiveness remains challenging. We describe the diagnostic performance evaluation of 3 common commercial MERS-CoV rRT-PCR assays on a large panel (n = 234) of upper respiratory tract specimens collected during an outbreak episode in Saudi Arabia. Assays were compared to the RealStar® MERS-CoV RT-PCR (Alton Diagnostics, Hamburg, Germany) assay as the gold standard. Results showed i) the TIB MolBiol® LightMix UpE and Orf1a assays (TIB MolBiol, Berlin, Germany) to be the most sensitive, followed by ii) the Anyplex™ Seegene MERS-CoV assay (Seegene, Seoul, Korea), and finally iii) the PrimerDesign™ Genesig® HCoV_2012 assay (PrimerDesign, England, United Kingdom). We also evaluate a modified protocol for the PrimerDesign™ Genesig® HCoV_2012 assay., Highlights • Diagnostic performance evaluation of 3 commercial MERS-CoV rRT-PCR assays. • LightMix UpE/Orf1a, Seegene MERS-CoV, and Genesig HCoV_2012 assays were evaluated. • Two hundred thiry-four URT specimens collected during an outbreak episode in Saudi Arabia. • Assays were compared to the RealStar® MERS-CoV RT-PCR assay as the gold standard. • Results showed the TIB MolBiol LightMix UpE/Orf1a assays to be the most sensitive.