Isolation and Characterization of Cholera Toxin Gene-Positive Vibrio cholerae Non-O1/Non-O139 Isolated from Urinary Tract Infection: A Case Report.

Background: Urinary tract infection (UTI) caused by V. cholerae is rare and less common. V. cholerae is a Gram-negative bacterium motile using single polar flagellum and, originally, is a waterborne microbe found in aquatic and estuarine environments. Toxigenic V. cholerae is well-known as a causative agent of acute and excessive watery diarrhea after ingesting food and water contaminated with this bacterium. Case Presentation: A 27-year-old male patient presented to the emergency department on 17th July 2021 with burning micturition, normal vital signs, and no fever, vomiting, or diarrhea. In 2017, the patient complained of short stature and vitamin D deficiency. He was on human growth hormone from January 2018 till October 2019. The diagnosis was V. cholerae Non-O1/non-O139 urinary tract infection (UTI). Considering a urinary tract infection, empirical treatment with Lornoxicam and Ciprofloxacin was initiated, while the result of urine culture was still pending. The patient was discharged on the same day and without any complications. Conclusion: V. cholerae non-O1/non-O139 is primarily a marine inhabitant and is associated with sporadic cases resulting in cholera-like diarrhea after consumption of contaminated seafood and exposure to seawater. Extraintestinal infection associated with this bacterium should no longer be ignored as this change in the behavior of cholera bacteria mechanism of pathogenicity might be related to some associated virulence genes.

Serological and Antibiotic Resistance Patterns As Well As Molecular Characterization of Vibrio parahaemolyticus Isolated from Coastal Waters in the Eastern Province of Saudi Arabia.

Vibrio parahaemolyticus belongs to the halophilic genus of Vibrionaceae family that inhabits coastal and marine environments and is a major food-borne pathogen. In the Gulf Cooperation Council (GCC) countries and Saudi Arabia in particular, there is a lack of information regarding the detection of pandemic clone or serovariants of V. parahaemolyticus pandemic clones. Here, 400 seawater samples were collected and examined for the presence of V. parahaemolyticus from 10 locations along the coast of Eastern Province in Saudi Arabia. The recovered isolates were serotyped, and studied for antimicrobial resistance, virulence genes, and markers of pandemicity using PCR and Arbitrarily primed (AP)-PCR typing patterns. All 40 isolates were tested negative for tdh, trh, and toxRS genes. Six serotypes were identified and three were clinically significant. Antibiotic susceptibility testing of isolates revealed high resistance towards penicillins, cephalosporins, and polymyxin; 60% of isolates were multi-drug resistant, whereas all isolates were susceptible to quinolones, carbapenems, sulfonamides, and tetracycline. The multiple antibiotic resistance (MAR) index among antibiotic resistance patterns of isolates revealed that 12 (30%) isolates had recorded significant MAR index higher than 0.2. AP-PCR fingerprinting could group all isolates into five distinct and identical pattern clusters with more than 85% similarity. Our findings demonstrate that pandemic serovariants of pandemic clones were not exclusively limited to strains isolated from fecal specimens of infected patients. Nine environmental strains of serotype O1:KUT, O1: K25, and O5:K17 were isolated from costal seawater, and thus the spread of these serovariants strains of pandemic clone of V. parahaemolyticus in the environment is to avoid any kind of threat to public health.

Molecular characterization of Helicobacter pylori clinical isolates from Eastern Saudi Arabia.

OBJECTIVES: To describe the frequency of cytotoxin-associated gene A (CagA) and vacuolating cytotoxin A (VacA) virulence genes and clarithromycin resistance-associated mutations among Helicobacter pylori (H. pylori) clinical isolates from Eastern Saudi Arabia. METHODS: A cross-sectional study was carried out between July 2020 and June 2021 in a tertiary hospital in AL-Khobar, Saudi Arabia. A total of 34 H. pylori isolates were obtained from gastric biopsies of patients with dyspepsia. The existence of the virulence genes was studied by polymerase chain reaction and the gene fragment of the 23s ribosomal subunit (23s rRNA) gene was sequenced. RESULTS: All isolates harbored the CagA gene. Approximately 97.1% (33/34) isolates were positive using the VacA M primer and 91.2% (31/34) isolates were positive using the VacA S primer. The most frequent allelic combination was S2/M2/cag (60%), followed by S1/M2/cag (26.7%), S1/M1/cag (10%), and S2/M1/cag (3.3%). Approximately 6.5% isolates harbored the A2142G mutation and 29% isolates harbored the A2143G mutation. One isolate contained the mutation T2182C. The phylogenetic analysis showed that 58% isolates clustered with the regional and global isolates while the remaining 42% isolates seemed to be specifically circulating in Saudi Arabia. Most of the patients (73.5%) had already underwent a previous H. pylori eradication therapy. CONCLUSION: We showed that there is a regional variation in the frequency of the virulence genes among H. pylori isolates. Additionally, we showed the frequency of 23s rRNA mutations related to clarithromycin resistance in Saudi Arabia.

Molecular epidemiology of SEN virus among blood donors and renal dialysis patients.

BACKGROUND AND PURPOSE: The SEN virus (SEN-V) is a single-stranded circular, non-enveloped DNA virus that has been linked to blood transfusion and is thought to be a major cause of post-transfusion hepatitis. The two SENV types, SENV-H and SENV-D, are non-A to E hepatitis viruses  in those who are infected. The purpose of this study is to find out how common SENV and its variations are among renal dialysis patients and healthy blood donors. METHODS: The study used a cross-sectional design, with 300 blood samples collected from KFMMC patients, 150 from healthy blood donors and 150 from renal dialysis patients, between January 2019 and January 2021. The samples were screened for the presence of SENV-D and SENV-H. using nested PCR. RESULTS: Molecular analysis of the SEN virus revealed that 9.3% of the samples (14 out of 150) tested positive for SEN virus infection in renal dialysis patients. The data from healthy donors revealed that 10% of the samples tested positive for the SEN virus (15 out of 150). CONCLUSIONS: The presence of SEN-V in healthy blood donors and renal dialysis patients demonstrates the virus's blood-borne nature and emphasizes the dangers of blood-borne transmission.

Monkeypox and Its Possible Sexual Transmission: Where Are We Now with Its Evidence?

Monkeypox is a rare disease but is increasing in incidence in different countries since the first case was diagnosed in the UK by the United Kingdom (UK) Health Security Agency on 6 May 2022. As of 9 August, almost 32,000 cases have been identified in 89 countries. In endemic areas, the monkeypox virus (MPXV) is commonly transmitted through zoonosis, while in non-endemic regions, it is spread through human-to-human transmission. Symptoms can include flu-like symptoms, rash, or sores on the hands, feet, genitalia, or anus. In addition, people who did not take the smallpox vaccine were more likely to be infected than others. The exact pathogenesis and mechanisms are still unclear; however, most identified cases are reported in men who have sex with other men (MSM). According to the CDC, transmission can happen with any sexual or non-sexual contact with the infected person. However, a recent pooled meta-analysis reported that sexual contact is involved in more than 91% of cases. Moreover, it is the first time that semen analysis for many patients has shown positive monkeypox virus DNA. Therefore, in this review, we will describe transmission methods for MPXV while focusing mainly on potential sexual transmission and associated sexually transmitted infections. We will also highlight the preventive measures that can limit the spread of the diseases in this regard.

Molecular Epidemiological Surveillance of CTX-M-15-producing Klebsiella pneumoniae from the patients of a teaching hospital in Sindh, Pakistan.

Background  The presence of Extended-spectrum ß-lactamase (ESBL) positive bacteria in hospital setting is an aggravating influential factor for hospitalized patients, and its consequences may be hazardous. Therefore, there is a need for rapid detection methods for newly emerging drug-resistant bacteria. This study was aimed at the molecular characterization of ESBL-positive  Klebsiella pneumoniae isolates recovered from clinical samples.   Methods  A total of 513  K. pneumoniae isolates were obtained from various clinical samples during June 2019 to May 2020. The collected isolates were investigated for antimicrobial susceptibility (antibiogram), and PCR and DNA sequencing were performed to analyse the ESBL genes.   Results  Among the 513 isolates, as many as 359 (69.9%) were ESBL producers and 87.5% were multi-drug resistant, while none had resistance to imipenem. PCR scored 3% blaTEM, 3% blaSHV, and 60% blaCTX-M-15 genes for the tested isolates.   Conclusion  The study showed that CTX-M-15 was the major prevalent ESBL type among the isolates. Additionally, all the isolates were susceptible to carbapenems. Screening and detection of ESBL tests are necessary among all isolates from the enterobacteriaceae family in routine microbiology laboratory to prevent associated nosocomial infections. A larger study is essential to understand molecular epidemiology of ESBL producing organisms to minimize morbidities due to these multidrug resistant organisms.

Comparative cross-sectional assessment of knowledge, attitude and practice among university students and employees towards the use of the microbiology laboratory equipment.

Background: Continuous evaluation of students and employee's knowledge and attitude in clinical laboratories is mandatory to ensure a high level of competency, proper practice and to assess the need for training, which shall be reflected on the quality of laboratory results. The aim of the present study was to evaluate the knowledge, attitude, and practice in microbiology laboratories among employees (at King Fahd Hospital of the University) and clinical laboratory students (at Imam Abdulrahman Bin Faisal University)    Methods: This was a cross-sectional survey of 30 2 nd year students, 26 3 rd year students, 24 4 th year students in the Clinical Laboratory Sciences department, and 30 employees. Participants completed a survey comprising 30 questions to assess their knowledge and attitude towards the use of equipment and practice in the microbiology laboratory. Results: The results indicated that there was no significant difference between the average scores of all levels of students regarding their knowledge (p = 0.85, 0.999, and 0.869), attitude (p = 0.883, 0.996, 0.853), and practice (p=0.633, 0.325, 0.858) in the microbiology laboratory. Employees scores (knowledge;5.03±2.646, attitude; 12.03±4.89, and practice; 7.7±6.11) were quite poor, as indicated by the lower average results than that of students (knowledge; 5.65±3.08, attitude; 13.25±5.33, and practice; 13.46±5.7). Conclusions: It is concluded that the knowledge, attitude, and practice of students and employees in the microbiology laboratory needs to be meticulously monitored and improved to ensure high achievement of learning outcomes and better overall performance in the laboratory. This may be achieved through using frequent quizzes and continuous education programs.

Direct DNA Sequencing-Based Analysis of Microbiota Associated with Hematological Malignancies in the Eastern Province of Saudi Arabia.

INTRODUCTION: Bloodstream infections (BSI) among patients with hematological malignancies (HM) could predispose them to higher morbidity and mortality for various underlying conditions. Several microorganisms, either pathogenic or opportunistic normal human flora, could cause severe bacteremia and septicemia. While conventional methods have their own limitations, molecular methods such as next-generation sequencing (NGS) can detect these blood infections with more reliability, specificity, and sensitivity, in addition to information on microbial population landscape. Methodology. Blood samples from HM patients (n = 50) and volunteer blood donor control individuals with no HM (n = 50) were subjected to 16S rRNA gene amplification using standard PCR protocols. A metagenomic library was prepared, and NGS was run on a MiSeq (Illumina) sequencer. Sequence reads were analyzed using MiSeq Reporter, and microbial taxa were aligned using the Green Genes library. RESULTS: 82% of the patients showed BSI with Gram-negative bacteria as the most predominant group. E. coli comprised a major chunk of the bacterial population (19.51%), followed by K. pneumoniae (17.07%). The CoNS and Viridans Streptococci groups are 17.07% and 14.63%, respectively. Other major species were S. aureus (9.75%), P. aeruginosa (7.31%), A. baumannii (4.87%), E. cloacae (4.87%), and P. mirabilis (4.87%). 34.14% of the cases among patients showed a Gram-positive infection, while 14.63% showed polymicrobial infections. CONCLUSION: Most of the BSI in patients were characterized by polymicrobial infections, unlike the control samples. Molecular methods like NGS showed robust, fast, and specific identification of infectious agents in BSI in HM, indicating the possibility of its application in routine follow-up of such patients for infections.

Antibiotic resistance and molecular characterization of enteroaggregative Escherichia coli isolated from patients with diarrhea in the Eastern Province of Saudi Arabia.

AIM: To investigate presence of Enteroaggregative Escherichia coli (EAEC) in patients suffering with diarrhea by targeting the pCVD432 (pAA) gene using PCR. METHODS: There were 63 non-duplicate isolates of E. coli isolated from diarrheal cases in teaching hospital in Eastern Province of Saudi Arabia between May 2013 to July 2014. All E. coli strains were examined for antibiotic susceptibility testing and polymerase chain reaction (PCR) for detection of virulence gene markers for EAEC. RESULTS: Of the 63 E coli strains that were reported with diarrheal cases, 35 (55.6%) EAEC were tested positive for pCVD432 gene and aggR gene was present in 19 (54.3%) strains. All strains tested positive for pCVD432 and aggR genes were classified as typical EAEC (tEAEC). EAEC revealed resistance to tetracycline, ampicillin, nalidixic acid, trimethoprim sulfamethoxazole, ciprofloxacin, streptomycin, noroxin, and piperacillin. CONCLUSION: EAEC was detected for the first time, among Saudi patients with diarrhea in this region of Saudi Arabia. The reported antibiotic resistance in this study is considered high among isolated EAEC strains to routinely prescribed antibiotics in our area.

Reporting Clinical Laboratory Critical Values: A Focus On The Recommendations Of The American College Of Pathologists.

BACKGROUND: Reporting critical values continues to receive a widespread attention from health care givers as it symbolizes a crucial clinic-laboratory link. This is because healthcare providers did realize the importance of prompt and timely communication of the critical results, which have positive implications on patient safety and treatment outcomes. In addition to physician and nurses, patients are also recipients of critical values, where they can make informed decisions prior to clinical intervention. The College of American Pathologists (CAP) requires stringent policies for reporting critical values in all laboratories, including adoption of a robust quality assurance system. Research studies have indicated that there still no universally accepted critical values list. This is because of various factors; such as differences in institutional organization, patient population, clinical demand, staffing, and instrumentation. However, through collaboration with other stakeholders involved in the delivery of healthcare, lab professionals may be able to come up with a realistic critical values list that reflects on the local needs and dynamics of patients' service. CONCLUSIONS: This review offers an insight into the process of reporting critical values, some challenges encountered, as well as the policies and procedures of effective reporting with a particular focus on the guidelines of the College of American Pathologists. There should be a common global guideline introduced by the health care governing agencies to be adapted with some flexibility in clinical laboratories in different clinical settings.

Prevalence of Hepatitis B Virus (HBV) Among Blood Donors in Eastern Saudi Arabia: Results From a Five-Year Retrospective Study of HBV Seromarkers.

BACKGROUND: Transfusion-transmissible hepatitis B virus (HBV) infection is a major problem worldwide. Recently, confirmatory nucleic acid tests (NATs) for HBV DNA have been employed in several countries. We assessed the prevalence and yearly trends of HBV infection in blood donors in the Eastern Province of Saudi Arabia, screening for HBV surface antigen (HBsAg), antibody against HBV core antigen (anti-HBc), and HBV DNA. METHODS: Between 2011 and 2015, a total of 22,842 donors were screenedfor HBsAg, anti-HBc, and HBV DNA using the HBsAg Qualitative II kit (Abbott, Ireland Diagnostics Division, Sligo, Ireland), ARCHITECT Anti-hepatitis B core antigen antibody (HBc) II Assay kit (Abbott GmbH & Co. KG, Wiesbaden, Germany), and NAT Procleix Ultrio Elite Assay kit (Grifols Diagnostic Solutions Inc., Los Angeles, CA, USA), respectively. RESULTS: A total of 739 (3.24%) donors were HbsAg(+), anti-HBc(+), or HBV DNA(+); 63 (0.28%) were HbsAg(+), anti-HBc(+), and HBV DNA(+). Twelve (0.05%) were anti-HBc(+) and HBV DNA(+) but HBsAg(-); they were considered to have occult infection. Further, 664 (2.91%) were HBsAg(-) but anti-HBc(+), indicating chronic or resolving infection. HBV prevalence increased significantly from 2011 to 2012, increased marginally till 2013, and showed a decreasing trend from 2013 (P>0.05). CONCLUSIONS: The five-year prevalence of HBV infection among blood donors in the Eastern Province of Saudi Arabia (3.24%) is lower than that reported for other regions in the country. The occult HBV infection rate of 0.05% emphasizes the importance of NATs in isolating potential infectious blood units.

Prevalence of digestive tract colonization of carbapenem-resistant Acinetobacter baumannii in hospitals in Saudi Arabia.

Carbapenem-resistant Acinetobacter baumannii is a major health problem worldwide, especially in intensive care units (ICUs). This study aimed to detect the prevalence of A. baumannii colonization of the gastrointestinal tract of patients admitted to the ICU in two hospitals in Saudi Arabia. In addition, it aimed to characterize the molecular mechanisms of carbapenem resistance in these isolates. From January to June 2014, 565 rectal swab specimens were screened for Acinetobacer strains and carbapenem resistance using CHROMagar Acinetobacter and CHROMagar KPC agar plates, respectively. Organism identification and susceptibility were detected using the Vitek 2 system. A total of 47 Acinetobacter spp. were detected, and 35 were resistant to carbapenem, making the prevalence of Acinetobacter spp. 8.3% (47/565) and carbapenem resistance (6.2%, 35/565). The 47 strains showed remarkable clonal diversity as revealed by PFGE. Using PCR, OXA-51, a chromosomal marker for A. baumannii, was detected in 46 strains. OXA-23 ß-lactamase was detected in all 35 carbapenem-resistant A. baumannii. No IMP, VIM, SPM, SIM, GIM, KPC or NDM ß-lactamases were detected in these isolates. Thus, OXA-23 was the main mechanism of carbapenem resistance in these isolates. To the best of our knowledge, this is the first study to detect the prevalence of Acinetobacter colonization in the digestive tract of ICU patients in Saudi Arabia. This study revealed the importance of having well-established protocols for early identification of these multidrug-resistant organisms, optimizing infection-control strategies and having active surveillance studies to reduce morbidity, mortality and cost.