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Objectives: Cutaneous leishmaniasis (CL) is considered an overlooked public health threat in KSA. CL is endemic to several Saudi regions, including Al Ahsa City, Eastern KSA. To our knowledge, no prior molecular identification of Leishmania parasites in vectors in Al Ahsa has been published. The aim of this study was to perform molecular identification of Leishmania species in sand flies collected from Al Ahsa., Methods: This cross-sectional study was conducted in Al Ahsa from July 2020 to May 2021. Sand flies were collected from the three areas with the highest rates of CL cases and sand flies, according to data from the Vector Borne Diseases Prevention Center in Al Ahsa. CDC miniature light traps and sticky traps were used to collect sand flies. Pools of 3-10 female sand flies were subjected to genomic DNA extraction. The extracted DNA was then amplified with a protocol targeting the Leishmania genus and using species-specific primers. For Leishmania species identification, a PCR-restriction fragment length polymorphism (PCR-RFLP) method was used., Results: Ten of 113 pools of samples tested positive for the Leishmania genus, according to our experimental protocol. Characterization of Leishmania species by PCR-RFLP established Leishmania major as the only species found in the collected sand flies., Conclusion: This is the first documentation of molecular identification of Leishmania species in phlebotomine sand flies in the Al Ahsa region. L. major was the only species identified in our study. Further comprehensive research investigating the vectors and reservoirs will be crucial to establish the dynamics of transmission of Leishmania in Al Ahsa., (© 2023 [The Author/The Authors].)

During the coronavirus disease 2019 lockdown period, a surge in sandflies and cutaneous leishmaniasis (CL) cases was observed in Al-Ahsa, Saudi Arabia. Skin punch biopsies were obtained from 100 patients clinically diagnosed with CL in Al-Ahsa who had no travel history in the last 6 months. Impression smears were used following a three-step polymerase chain reaction (PCR) protocol using genus-specific primers targeting kDNA and ITS1. Leishmania speciation was determined by ITS1 PCR/nested PCR-restriction fragment length polymorphism and sequencing. A phylogenetic tree was constructed. The associated patient characteristics were analyzed. Using internal transcribed spacer one (ITS1)-PCR/nested PCR, 98 cases were considered true-positive CL. Leishmania major was the predominant species, and Leishmania tropica was identified in three cases. Microscopy had poor sensitivity and perfect specificity. Direct ITS1-PCR missed nine cases. Sex, residence, and treatment outcome were significantly associated with the occurrence of Leishmania; distribution of skin lesion(s) and treatment outcome were significantly associated with Leishmania genotype. This is the first time that L. tropica was identified as a cause of CL in human in Al-Ahsa, in addition to the predominant zoonotic species, L. major. We recommend using ITS1-nested PCR for negative cases by ITS1-PCR. Further exploration of Leishmania transmission dynamics in vectors and reservoir animals is essential for designing effective preventive measures., (© 2022. The Author(s).)

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., (Copyright © 2021 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

The outbreak of the new human coronavirus SARS-CoV-2 (also known as 2019-nCoV) continues to increase globally. The real-time reverse transcription polymerase chain reaction (rRT-PCR) is the most used technique in virus detection. However, possible false-negative and false-positive results produce misleading consequences, making it necessary to improve existing methods. Here, we developed a multiplex rRT-PCR diagnostic method, which targets two viral genes (RdRP and E) and one human gene (RP) simultaneously. The reaction was tested by using pseudoviral RNA and human target mRNA sequences as a template. Also, the protocol was validated by using 14 clinical SARS-CoV-2 positive samples. The results are in good agreement with the CDC authorized Cepheid`s Xpert® Xpress SARS-CoV-2 diagnostic system (100%). Unlike single gene targeting strategies, the current method provides the amplification of two viral regions in the same PCR reaction. Therefore, an accurate SARS-CoV-2 diagnostic assay was provided, which allows testing of 91 samples in 96-well plates in per run. Thanks to this strategy, fast, reliable, and easy-to-use rRT-PCR method is obtained to diagnose SARS-CoV-2., Competing Interests: The authors have declared that no competing interests exist.

Brucellosis is considered a prevailing endemic infectious disease in the Kingdom of Saudi Arabia and represents a health problem with socio-economic burden. There are two main Brucella species that cause human brucellosis; Brucella abortus , and Brucella melitensis . The clinical features range from asymptomatic to the acute symptoms of fever, joint pain, muscle pain, headache, nausea/vomiting, anorexia and malaise in addition to the subsequent complications that might occur. The endemicity of brucellosis might be explained due to obstacles in controlling the importation of animals for slaughtering during Hajj periods and for several other predisposing factors. The distribution of the disease is all over the country and the most prevalent part is the south followed by north and then the east and central parts. However, in the complexity of brucellosis control measures, there are several activities which have been implemented to tackle the disease such as mass vaccination of animals, regulating importation of slaughter animals, and improving public awareness. This review provides a detailed description of the status of brucellosis in Saudi Arabia, which includes epidemiology, clinical characteristics, virulence and pathophysiology, and prevention of the disease., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2020 The Author(s).)

The current study proposes a bio-directed approach for the formation of titanium oxide and silver nanoparticles (TiO 2 and Ag NPs), using a wild mushroom, Fomitopsis pinicola , identified by 18S ribosomal RNA gene sequencing (gene accession no. MK635350) and phenotypic examination. NP synthesis was confirmed by X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FT-IR), diffuse reflectance UV-visible spectroscopy (DR-UV), and scanning and transmission electron microscopy (SEM/TEM). Furthermore, the impact of NPs on Escherichia coli and Staphylococcus aureus and a human colon cancer cell line (HCT) were evaluated by MIC/MBC and MTT assays, respectively, along with structural morphogenesis by different microscopy methods. The results obtained showed that TiO 2 and Ag NPs were found to be significantly active, however, slightly enhanced antibacterial and anticancer action was seen with Ag NPs (10-30 nm). Such NPs can be utilized to control and treat infectious diseases and colon cancer and therefore have potential in a range of biomedical applications., Competing Interests: No potential conflict of interest is reported by authors., (This journal is © The Royal Society of Chemistry.)

The establishment of a benign system for the nanoparticle (NPs) synthesis, is a key in nanotechnology for the environmental and health care industries. Therefore, enrichment of novel biological systems for the green synthesis is in significant demand, to lift up these compounds in the biomedical industries. The present work, reports the green synthesis of ZnO NPs, employing a novel thermophile, identified as Bacillus haynesii (GeneBank : MG822851) isolated from the leaf of date palm plant ( Phoenix dactylifera ), as an eco-friendly nanobiofactory. Physiochemical characterization of ZnO NPs (50 ± 5 nm in size), was achieved by Fourier transform infrared spectroscopy (FT-IR), X-ray diffraction (XRD), diffuse reflectance UV-Visible spectroscopy (DR UV-Vis spectroscopy), Thermogravimetry analysis (TGA), scanning electron microscopy (SEM) and transmissiom electron microscopy (TEM). The morphogenesis and antimicrobial activity of synthesized ZnO NPs, was studied by evaluating the minimum inhibitory/bactericidal concentration (MIC&MBC) against Escherchia coli (8 and 16 mg/mL) and Staphylococcus aureus (4 and 8 mg/mL), respectively. The present study encourages the use of B. haynesii for the green synthesis of ZnO NP. To the best of our knowledge, this is the first report on the study of thermophilic, B. haynesii for green synthesis of NPs in general and ZnO NPs in particular.

Background: Identification and assessment of therapeutic potential of natural products derived from medicinal plants have led to the discovery of innovative and economical drugs to treat several diseases, including chronic wounds. In vitro cell based scratch assay is an appropriate and inexpensive method for initial understanding of wound healing potential of medicinal plant extracts. The current study was aimed at investigating the wound healing capacity of Aristolochia saccata leaf extract by using scratch assay as a primary model, where proliferative and migratory capabilities of test compounds could be monitored through microscopy studies. A. saccata is an evergreen climbing shrub belongs to the family Aristolochiaceae., Methods: Methanolic extraction of the plant material was done using Soxhlet apparatus and the cytotoxicity of the extract on L929 cells was studied by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. L929 is a human fibroblast cell line. In vitro scratch assay was performed to evaluate the wound healing properties of A. saccata leaf extract and possible mechanism of action was analyzed by flow cytometric expression studies of an extracellular matrix (ECM) factor, collagen type-1., Results: MTT assay revealed that A. saccata leaf extract had no cytotoxic effect on the cells and at higher concentrations, the extract showed mild toxicity resulting in the death of just 2.88% cells. Scratch assay showed 34.05%, 70.00%, 93.52% wound closure at 12hrs, 24hrs and 48hrs of incubation respectively. These results were similar compared to positive control which showed 37.60, 56.41 and 99.05% of wound closure. Further, flow cytometry-based studies revealed that the A. saccata leaf extract induced the expression of ECM remodelling factor collagen-1., Conclusion: Our study revealed the wound healing capabilities of A. saccata In vitro . Hence, A. saccata could be recommended as a potential source of wound healing agents.