Introduction: The National Public Health Reference Laboratory at the National Center for Disease Control (NCDC), Tripoli was the single site for SARS-CoV-2 diagnostic testing in Libya. During the pandemic, large quantities of waste were generated each day. In response, the laboratory biosafety team conducted an assessment and implemented a plan to strengthen biosafety and biosecurity measures to mitigate high-risk biological waste.Objectives: To assess the potential biosafety and biosecurity hazards during surges in diagnostic response and create training programs and accompanying effective standard operating procedures (SOPs) that align with medical practices, ethical standards, available resources, and local regulations.Methods: A biosafety team was created to assess existing practices, identify gaps, and provide recommendations. SOPs were developed for biological waste collection, segregation, offsite transportation, treatment, and disposal.Results: From May 2020 to May 2022 approximately 22.8 kilograms of COVID-19 testing related laboratory waste was created and managed on daily basis. In response, NCDC's biosafety team achieved a number of key goals: All laboratory and contract employees were trained on waste management best practices; four waste management SOPs were developed and deployed to provide standard guidance; training sessions were conducted to enhance safety measures; biomedical waste were professionally managed, ensuring compliance with health and environmental standards; and the biosafety team conducted regular monitoring and inspections for accountability and adherence to protocols.Conclusion: Examining the complete lifespan of biological waste from creation to disposal can inform more comprehensive policy development and improve overall laboratory biosafety during pandemic response.

Cantharellus cibarius, sometimes known as the "boreal chanterelle", is a member of the phylum Basidiomycota and one of the most sought edible mushrooms in the world. This study compared the use of supercritical fluid extraction (SFE), which is regarded as a mild and green extraction method, with traditional solvent extraction to obtain C. cibarius oils. The optimized parameters of the SFE were obtained through a Box-Behnken design, and the SFE approach provided oil yields that exceeded conventional extraction yields with ethyl acetate (10.1% w/w vs 8.1% w/w) while exhibiting better selectivity for the accumulation of lipophilic bioactives. Most importantly, the supercritically extracted oil showed significant radical scavenging activities (65% inhibition against ABTS, compared to 58% in the conventionally ethyl acetate-extracted oil), antioxidant and anti-inflammatory properties, as well as bactericidal effects against Bacillus cereus, Escherichia coli, and Salmonella typhi. The C. cibarius oils extracted through a green technology could be a potential application as condiments with retained bioactive properties.

Acinetobacter baumannii is a major nosocomial pathogen, particularly in intensive care units (ICUs), due to its remarkable resistance to multiple antibiotics and its persistence in hospital environments. The occurrence of carbapenem-resistant A. baumannii, often driven by the metallo-β-lactamase genes such as blaIMP and blaVIM, results in a serious clinical threat. In this study, 119 A. baumannii samples were collected from different hospitals in Tripoli, Libya. Real-time PCR revealed that 29 (24.3%) isolates carry the blaIMP gene and 62 (52%) have the blaVIM gene. Phenotypic detection of carbapenemase production revealed that chromogenic media had the highest detection rate (74.7%), followed by the Modified Hodge Test (67%) and E-test (47%). Antimicrobial susceptibility testing showed 100% resistance to ertapenem, cefoxitin, ceftriaxone, ampicillin, and amoxicillin-clavulanate, with high resistance also observed against imipenem (88.2%), meropenem (88.2%), gentamycin (94.9%), ciprofloxacin (94%), and levofloxacin (92.4%). Colistin was the only antibiotic to which all A. baumannii isolates remained sensitive. These findings underscore the high burden of multidrug resistance and the widespread presence of blaVIM and blaIMP genes in A. baumannii isolates in Libya. Improved molecular surveillance, reliable detection methods, and improved antibiotic stewardship are urgently needed to control the spread of these resistant pathogens.

The growing demand for truffles has led to a global pursuit for their occurrence in the wild, and studies concerning their cultivation. Although European countries such as Italy, France and Spain have been known for truffle production, truffle hunting is new to Finland. The present study reports for the first time, the findings of Tuber maculatum in Finland on the basis of morphological and molecular analysis. The chemical characteristics of soil samples collected from the truffle sites have also been discussed. The species of the Tuber samples were identified primarily using morphological analysis. Molecular analysis was carried out for the confirmation of the identity of the species. Two phylogenetic trees were constructed based on internal transcribed spacer (ITS) sequences produced in this study and including representative sequences of whitish truffles available in GenBank. The truffles were identified as T. maculatum and T. anniae. This study could be considered as a basis for encouraging research on findings and identification of truffles in Finland.

Background:People are exposed to lead from car exhaust fumes (leaded petrol), lead-based paints, plumbing systems, cigarette smoking, contaminated soil and dust, foods grown in polluted areas, and water sources. Among the vulnerable subpopulations are pregnant women, fetuses and infants.Aims:To estimate and correlate maternal and neonatal blood lead levels and compare these between neonates and their mothers in Tripoli and Ghadames.Methods:In this cross-sectional study, blood was collected from 120 and 116 mothers and their neonates from Tripoli and Ghadames, respectively. Lead levels were determined using atomic absorption spectrophotometry. A simple questionnaire was filled by the participants using face-to face interview. Data were analysed using SPSS version 20.0 and Excel.Results:Mean blood and umbilical cord lead levels were 6.83 (standard deviation 4.96) and 6.05 (4.89) µg/dl in mothers and neonates from Tripoli, respectively, and 5.91 (4.02) and 4.54 (4.09) µg/dl from Ghadames. There was no significant difference in blood lead level between mothers from Tripoli and Ghadames. However, there was a significant correlation between maternal blood and umbilical cord blood in Tripoli and Ghadames. Linear regression revealed that neonatal umbilical cord blood lead levels reflected the levels in maternal blood. Blood lead in this study was higher than that reported in industrialized western countries.Conclusion:We detected moderate blood lead levels among pregnant women in Tripoli and Ghadames. It is important to detect the source of lead in the Libyan population to treat this problem effectively.

Abstract Angiogenesis is a complex physiological process that cannot be treated with single agent therapy. Several edible fungi have been known to encompass bioactive compounds, and are promising sources of multi-component drugs. One such widely consumed edible fungi is Cantharellus cibarius, which has been explored for its biological activities. The present study focused on assessing the anti-angiogenic activity of petroleum ether and ethanol extracts of C. cibarius using chick chorioallantoic membrane (CAM) assay. Both the extracts showed a dose-dependent response which was compared with the anti-angiogenic activity of the positive controls silibinin, and lenalidomide. The extracts were also studied for their lipoxygenase (LOX) inhibitory potential and compared to ascorbic acid as the positive control. The IC50 values of the petroleum ether extract, ethanol extract, and ascorbic acid for LOX inhibition assay were 135.4, 113.1, and 41.5 µg/mL, respectively. Although both the extracts showed similar responses in CAM assay, ethanol extract proved to be more potent in LOX inhibition assay. Finally, the extracts were investigated for their chemical composition using GC-MS. A correlation between LOX inhibition and anti-angiogenic potential was established at the molecular level. A meticulous literature search was carried out to correlate the biochemical composition of the extracts to their anti-angiogenic activity.

Background Vitamin D is a fat-soluble hormone that plays an important role in glycaemic control. In addition, it has a positive effect on improving liver enzyme function. Aim This study was performed to examine the effect of intramuscular injection of vitamin D on serum 25-hydroxyvitamin D [25(OH)D] levels, glycemic control, and liver enzymes in Libyan patients suffering from type 2 diabetes mellitus (T2DM) with vitamin D deficiency. Methods and Materials This cross-sectional study enrolled 100 T2DM (50 males and 50 females). Their serum 25(OH)D, fasting blood glucose (FBG), and liver enzymes were measured at the baseline and 12 weeks after treatment with vitamin D (200,000 IU) injection monthly for 3 months. Data analysis involved the estimation of mean ± standard error (SE) and comparison of means between pre and post-treatment values using paired t-test. Independent t-test was used to compare the means between males and females. The level of significance was set at p < 0.05. Results Females had a lower 25(OH)D blood levels than males at baseline (7.03 ± 0.25 ng/mL versus 7.86 ± 0.26 ng/mL, respectively p < 0.02). 25(OH)D levels in both sexes was increased significantly from 7.45 ± 0.18 ng/mL to 26.69 ± 0.24 ng/mL after 12 weeks of vitamin D injections (p < 0.001), with no significant differences between male and females. FBG levels in both sexes was significantly decreased from 144.68 ± 1.84 mg/dL to 85.96 ± 0.34 mg/dL post treatment (p < 0.001). Alanine aminotransferase (ALT) was increased from 10.24 ± 0.17 U/L at baseline to 20.34 ± 1.15 U/L post treatment (p < 0.001). Similarly, aspartate aminotransferase (AST) was increased from 11.23 ± 0.21 to 20.57 ± 0.22 U/L (p < 0.001), and alkaline phosphatase (ALP) was decreased from 124.95 ± 1.15 U/L to 111.17 ± 1.27 U/L (p < 0.001). There were no significant differences between male and female liver enzymes either pre- or post-vitamin D injections Conclusion Treatment with vitamin D injection showed a significant increase in 25(OH)D accompanied by decreased FBG and ALP levels and increased ALT and AST levels. Vitamin D levels should be monitored and adjusted in diabetic patients.

The COVID-19 epidemic started in Libya in March 2020 and rapidly spread. To shed some light on the severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) strains circulating in Libya, viruses isolated from 10 patients in this country were sequenced, characterized at the genomic level, and compared to genomes isolated in other parts of the world. As nine genomes out of 10 belonged to the SS1 cluster and one to SS4, three datasets were built. One included only African strains and the other two contained internationally representative SS1 and SS4 genomes. Genomic analysis showed that the Libyan strains have some peculiar features in addition to those reported in other world regions. Considering the countries in which the strains are genetically more similar to the Libyan strains, SARS-CoV-2 could have entered Libya from a North African country (possibly Egypt), sub-Saharan Africa (e.g., Ghana, Mali, Nigeria), the Middle East (e.g., Saudi Arabia), or Asia (India, Bangladesh).

Background and Aims: The risk of malaria re-introduction to Libya is increasing due to immigration to Libya from sub-Saharan countries where malaria is endemic. This study was conducted to investigate the prevalence of malaria-positive cases among illegal immigrants in the southern region of Libya. Materials and Methods: Three hundred and three illegal immigrants, with a mean age of 25.78 ± 5.92 years, from 12 different countries were included. An enzyme-linked immunosorbent assay was used to detect the presence of serum malaria antibodies. Results: One hundred and ninety-five (64.36%) of the participants tested positive for malaria antibodies. One hundred and seventy-two of the positive cases arrived in Libya within 1–10 months. The highest number of positive cases (70) came from Ghana, followed by (40) from Niger and (39) from Bangladesh. Conclusion: Illegal immigration issue should be treated urgently, combined by increasing the surveillance activities of infectious diseases to prevent malaria re-introduction to Libya.

Background: Many epidemiological studies reported the association of Vitamin D deficiency and abnormal liver enzymes' levels with type 2 diabetes (T2D) and their findings remain inconsistent. Aim: The study investigated the status of both Vitamin D levels and liver enzymes' activity in a Libyan population with T2D. Materials and Methods: A total of 180 individuals of both genders whose ages ranged between 25 and 62 years were enrolled. Fasting blood sugar, (FBS), Vitamin D, aspartate aminotransferase (AST), alkaline phosphatase (ALP), and alanine aminotransferase (ALT) were measured in patients' serum. The association of both Vitamin D and liver enzymes levels with T2D was evaluated by Chi-square analyses. Results: 55.6% of the study subjects were diabetics and 44.4% were healthy controls. 100% of T2D patients had deficient Vitamin D level (<20 ng/ml) (P = 0.000). The mean Vitamin D level was higher in nondiabetics (39.1 ± 4.0) ng/ml than in T2D patients (7.4 ± 1.8) ng/ml. 98% (P = 0.068), 97% (P = 0.118), and 100% of T2D patients had normal levels of AST, ALP, and ALT enzymes, respectively. The mean AST, ALP, and ALT enzymes levels for nondiabetics were 30.3 ± 4.1, 58.7 ± 8.0, and 24.8 ± 4.9 U/L, respectively. The mean AST, ALP, and ALT enzymes levels for T2D patients were 11.3 ± 2.9, 125.0 ± 11.5, and 10.3 ± 1.7 U/L, respectively. Conclusions: Vitamin D deficiency may be related to T2D in the Libyan population, while the liver enzymes showed no significant differences between diabetics and nondiabetics.

Factor V Leiden G1691A (FVL) and Factor II prothrombin G20210A (PGM) mutations are the leading causes of thrombophilia. In this study, we have investigated the prevalence of the FVL G1691A and PGM G20210A single nucleotide polymorphisms (SNPs) among Libyan deep vein thrombosis (DVT) and myocardial infarction (MI) patients. SNP genotyping was performed using high-resolution melt analysis (HRM) and DNA sequencing. Biochemical parameters conducted on 112 males and 93 females showed no significant difference in means between the control group and the deep vein thrombosis and myocardial infarction groups. For Factor V Leiden, 40 samples were genotyped. Of the 40 samples, 6 (15.0%) of them were heterozygous and no one was homozygous. As for Factor II SNP, 59 samples were genotyped and only 2 (3.3%) were heterozygous. All the heterozygous samples showed 100% concordance between the HRM-PCR and DNA sequence analysis. Our study showed, for the first time, that both the FVL and PGM mutations are present among Libyan DVT and MI patients and that the FVL mutation is significantly associated with DVT but not with MI. However, our results do not support the association of PGM G20210A mutation with DVT or MI.

Cholesterol is probably the best known steroid because of its association with atherosclerosis. Previous studies shown association of olive oil use with reduced serum cholesterol levels. In addition, honey has been shown to have beneficial effects on lipid profiles. Therefore, this study was carried out to investigate the effects of olive oil and honey on total cholesterol level in healthy Libyan individuals. The study was conducted on 75 Libya volunteers aged 40-55 years divided into 3 groups including, 25 olive oil group aged 49.47±2.15, honey group aged 50.07±2.01, olive oil and honey group aged 52.07±4.31. The total cholesterol level was measured before and after the experiment and expressed as (mg/dL).The chemical and physical properties of the olive oil used in the study were found to be compatible with permitted limits issued by the Libyan National Centre of Standardization and Metrology .Our results showed that olive oil and honey significantly reduced total serum cholesterol (p<0.001) and (p<0.05) respectively. Furthermore, combination of olive oil and honey significantly reduced total cholesterol (p<0.001) by 21%. We conclude that the utilization of olive oil and honey can lead to a significant decrease in the levels of total serum cholesterol in blood.

Fungi are a huge source of unexplored bioactive compounds. Owing to their biological activities, several fungi have shown commercial application in the health industry. Tuber aestivum Vittad. is one such edible fungi with an immense scope for practical biological applications. In the present study, the anti-angiogenic activity of petroleum ether and ethanol extracts of T. aestivum was investigated using the chick chorioallantoic membrane assay and compared to the positive controls silibinin and lenalidomide. Both the extracts showed a dose-dependent anti-angiogenic response. The extracts were also assessed for their anti-inflammatory potential by lipoxygenase-inhibition assay. The IC50 values for LOX inhibition assay, computed by the Boltzmann plot, were 368.5, 147.3 and 40.2 mu g/mL, for the petroleum ether extract, ethanol extract, and the positive control ascorbic acid, respectively. The ethanol extract of T. aestivum showed superior anti-angiogenic and anti-inflammatory activity than the petroleum ether extract. Compositional investigation of the extracts by GC-MS revealed the presence of various bioactive compounds. The compounds were correlated to their anti-angiogenic and antiinflammatory activity based on a meticulous literature search.

Despite the relative abundance of sunny weather, surprisingly, there is increasing evidence that vitamin D deficiency is extremely prevalent in females of reproductive age in Middle East countries. There is also increasing interest in the non-classical roles of vitamin in health and disease including its relation to incidence of gestational diabetes, its impact on glycaemic control in diabetes mellitus, and its association with some complications of pregnancy like preeclampsia. The objective of this study was to estimate the prevalence of Vitamin D deficiency in pregnant diabetic patients in west Libya and analyse potential links to socioeconomic and cultural factors. This is a cross sectional observational study. Random plasma was collected form expected mothers attending the Antenatal Diabetes Clinic at Tripoli’s Main Maternity Hospital. Demographics and socioeconomic and cultural factors were recorded at the same time. Samples were analysed for vitamin D level and biochemical screening panel. Vitamin D level was obtained from 160 patients (mean age 35 years). Over all 95 % of the study population had vitamin D levels below normal (defined as vitamin D level of < 20 ng/mL). Results were subcategorised into severe deficiency (<10 ng/ml, 51.9%-83 patients), deficiency (< 20 ng/mL, 43.1%, 69 patients), insufficiency (20 - 30 ng/ml, 3.8%, 6 patients) and sufficient (> 30 ng/ml, only 1.3%, 2 patients). All patients were taking daily vitamin D Supplements at a dose of 400 IU as per hospital policy. Vitamin D deficiency is extremely prevalent in pregnant diabetic patients in Libya. There is no clear association with socioeconomic risk factors like employment, type of accommodation or geographic distribution. However, most of the study population had life style characterized by minimal exposure to direct sun light. Routine supplementation of Vitamin D in doses of 400 IU/day does not appear to ameliorate the severity of vitamin D deficiency in this group.

Warfarin is commonly prescribed as oral anticoagulant medication for Libyan patients, the wide inter-individual variation between the patients in their response to oral anticoagulants is attributed to genetics factors, mainly polymorphisms in CYP2C9 and VKORC1. This study was aimed to assess the impact of genetic (CYP2C9*2, *3 and VKORC1- 1639G>A/ and 1173 C>T polymorphism), and non- genetic factors: age, and body mass index (BMI) in the response of Libyan patients using oral anticoagulants.A total of 100 patients with stable maintenance dose of warfarin or acenocoumarol were recruited during their routine follow up in anticoagulant clinic at Tripoli Medical Centre. CYP2C9 and VKORC1 variant alleles were screened by (HRM) real-time PCR, followed by DNA sequencing.The variant allele frequencies of CYP2C9*2, CYP2C9*3, and VKORC1 -1639G>A/1173C>T were 9.5%, 4.0%, 4.5%, and 37.0%, respectively.Carriers of VKORC1 (-1639 G>A, 1173 C>T) variant alleles required a significantly lower doses of oral anticoagulants compared with carriers of wild type, P value =0.04, and 0.019, respectively. No significant difference in dose requirement was found between carriers of wild type, and CYP2C9*2 and *3 variant alleles, P value =0.11 and 0.98, respectively.The multivariate regression model including age, BMI, VKORC1, and CYP2C9 genotype produced weak model for estimating the drug dosage (R2= 8.6%); and neither genetic nor non-genetic factors could be used as a predictor for estimation of oral anticoagulant dosage.Our data showed that VKORC1 variant alleles but not CYP2C9*2, *3 variant alleles significantly contributed to oral anticoagulant dose variability.

Background and Aims: Despite the well-known association between human papillomavirus (HPV) and cervical cancer, yet there are no available data concerning the prevalence of HPV and its type distribution among Libyan women. The aim of this study was to investigate the prevalence of the most common high-risk HPV types 16 and 18 among Libyan women in Tripoli and to compare it with the cytological findings of the cervix. Methods: A total of 132 cervical samples were collected from women who sought medical attention at the gynecology outpatient clinic of the Tripoli University Hospital and other gynecology private clinics in Tripoli region. Cervical cytological status was classified according to the Bethesda System 2014. Quantitative polymerase chain reaction was used to facilitate the specific detection of HPV types 16 and/or 18. Results: The cytopathological examination showed that 92.4% of women had normal cervical cytology (n = 122/132) and 7.5% (n = 10/132) of them had cervical lesions. The overall prevalence of the most common oncogenic HPV types was 4.5%, as only six samples (n = 6/132) were confirmed of harboring HPV-DNA. Concerning the cytological status of the cervix, HPV-DNA was not found (0%) in women with a normal cervix, and it was present in 60% of women with cervical lesions. The high-risk HPV type 16 was the exclusive type among our all positive samples, with no detection of HPV type 18 among all our recruited subjects. Conclusion: Even though our findings showed a low overall prevalence of high-risk HPV types among Libyan women, the burden of HPV 16 among women with cervical lesions highlights the need to raise attention toward expanding research about HPV and adopt measures to prevent cervical cancer by vaccination and national screening program. The introduction of HPV-DNA testing in cervical cancer management will greatly benefit early-stage HPV detection and help prevent cervical lesions from progression to cancer.

OBJECTIVESMultidrug resistance (MDR) and emergence of extended-spectrum β-lactamases (ESBLs) among uropathogenic Escherichia coli have been reported worldwide, but there was no information on the detection of blaCTX-M-15 in major teaching hospitals in Libya. The aim of the study was to investigate the occurrence of CTX-M-15 β-lactamases producers isolated from five teaching hospitals in Tripoli, Libya.METHODSA total of 346 urine samples were collected from hospitalized patients in five teaching hospitals with a diagnosis of urinary tract infection (UTI). Phenotypic confirmation of ESBLs was confirmed by E-test strip; all ESBL-producing E. coli isolates were screened for the blaCTX-M-15 gene.RESULTSThe distribution of ESBL-producing E. coli varied among the five hospitals. The highest proportion was identified in Tripoli Medical Centre (67.6%). There were extremely high proportions of isolates resistant to ceftriaxone, cefepime, and ceftazidime (93.0-100.0%) among ESBL producers compared to non-ESBL producers (2.2-4.7%). MDR was detected in 22.2% of isolates. The majority of isolates (85.9%) in which blaCTX-M-15 was identified were ESBL producers. There was a correlation (p < 0.001) between expression of CTX-M-15 and resistance to ceftazidime.CONCLUSIONSThe isolation of MDR ESBL-producing uropathogens expressing the CTX-M-15 gene will limit the choices clinicians have to treat their patients with UTIs. Continued surveillance and implementation of efficient infection control measures are required.

Introduction: Extended-spectrum β-lactamases (ESBLs), including the AmpC type, are important mechanisms of resistance among Klebsiella pneumoniae and Escherichia coli isolates. Objective: The aim of the study was to investigate the occurrence of AmpC-type β-lactamase producers isolated from two hospitals in Tripoli, Libya. Methods: All clinical isolates (76 K. pneumoniae and 75 E. coli) collected over two years (2013-2014) were evaluated for susceptibility to a panel of antimicrobials and were analyzed phenotypically for the ESBL and AmpC phenotype using E-test and ESBL and AmpC screen disc test. Both ESBL and AmpC-positive isolates were then screened for the presence of genes encoding plasmid-mediated AmpC β-lactamases by polymerase chain reaction (PCR). Results: Of the K. pneumoniae and E. coli tested, 75% and 16% were resistant to gentamicin, 74% and 1.3% to imipenem, 71% and 12% to cefoxitin, 80% and 12% to cefepime, 69% and 22.6% to ciprofloxacin, respectively. None of the E. coli isolates were multidrug resistant compared with K. pneumoniae (65.8%). K. pneumoniae ESBL producers were significantly higher (85.5%) compared with (17.3%) E. coli isolates (P <0.0001, OR=4.93). Plasmid-mediated AmpC genes were detected in 7.9% of K. pneumoniae, and 4% E. coli isolates. There was low agreement between phenotypic and genotypic methods, phenotypic testing underestimated detection of AmpC enzyme and did not correlate well with molecular results. The gene encoding CMY enzyme was the most prevalent (66.6%) of AmpC positive isolates followed by MOX, DHA and EBC. Only one AmpC gene was detected in 5/9 isolates, i.e, blaCMY (n=3), bla MOX (n=1), blaDHA (n=1). However, co-occurrence of AmpC genes were evident in 3/9 isolates with the following distribution: bla CMY and blaEBC (n=1), and blaCMY and blaMOX (n=2). Neither blaFOX nor blaACC was detected in all tested isolates. All AmpC positive strains were resistant to cefoxitin and isolated from patients admitted to intensive care units. Conclusion: Further studies are needed for detection of other AmpC variant enzyme production among such isolates. Continued surveillance and judicious antibiotic usage together with the implementation of efficient infection control measures are absolutely required.