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Background: Acinetobacter baumannii is a notorious nosocomial pathogen universally in healthcare settings. Its natural competent characteristics for genetic recombination are responsible for acquired antibiotic resistance and render it untreatable through commonly used antibiotics. Hence, characterizing the A. baumannii genomes for multidrug resistance carriage is of paramount importance. The study aimed to characterize the whole genome of clinical isolates of A. baumannii to identify specifically the types of antibiotic resistance genes, drug classes and mobile genetic elements. We also aimed to determine the significant multi-locus sequence tags (MLSTs). The phylogeny of the isolates was established with other clinical strains distributed globally. Methods and results: Fifteen clinical isolates (isolated from tracheal secretion, urine and bronchoalveolar lavage) were subjected to whole genome sequencing. Raw sequences were assembled using SPAdes and species were identified using KmerFinder 3.2. The assembled genomes were annotated using the Prokka v1.14.6. Resfinder 4.6.0 was used to determine antibiotic resistance genes. The sequences were aligned against seven housekeeping genes aka sequence tags (STs) available within the MLST database (v 2.0.9). MobileGeneticElement finder (v1.0.3) were used for profiling mobile genetic elements associated with the antibiotic resistance genes. The genomes of nosocomial A. baumannii were assembled with an average N50 of 23,480 and GC content of 38%. There were approximately 3700 CDs, 53 tRNA and 3 rRNA. About 80% of the isolates were ST2 type. The genomes possessed antibiotic resistance genes (n = 24) belonging to 17 drug classes. The predicted phenotype was multidrug resistant. Among the mobile genetic elements, 12 insertion sequences and 2 composite transposons were also found. The mode of antibiotic resistance was mostly through antibiotic inactivation in all the isolates. Conclusions: The results imply the occurrence of multidrug resistant genes in clinical isolates of A. baumannii strains in the healthcare settings of Kuwait. A more comprehensive survey should be undertaken for antimicrobial resistance monitoring on a regular basis for surveillance, contact tracing, and potential mitigation in clinical settings. [ABSTRACT FROM AUTHOR] more...

Introduction: The northwest Arabian Gulf encounters significant anthropogenic pressures, including nutrient enrichment from coastal development and effluent discharge. Methods: This study presents the first shotgun metagenomics-based characterization of microbial communities in Kuwaiti waters of the northwest Arabian Gulf, focusing on Kuwait's first Marine Protected Area (MPA) in Sulaibikhat Bay, a vital nursery ground for commercially important fish. Results: Analysis revealed significantly higher microbial diversity within the MPA compared to adjacent waters, with Rhodobacteraceae (27.8%) and Flavobacteriaceae (15.3%) being dominant. Elevated inorganic phosphorus, nitrogen, and salinity were key factors driving this diversity. Multivariate analysis highlighted phosphate as a critical component affecting the MPA microbial community structure, particularly for the families Microbacteriaceae, Flavobacteriaceae, and Rhodobacteraceae. Discussion: This study underscores the ecological importance of MPAs and highlights the impact of nutrient enrichment and other environmental stressors on microbial diversity, emphasizing the need to reduce nutrient influx to mitigate eutrophication and enhance marine ecosystem resilience in stressed environments. [ABSTRACT FROM AUTHOR] more...

Fungi are an important part of the atmospheric ecosystem yet an underexplored group. Airborne pathogenic fungi are the root cause of hypersensitive and allergenic states highly prevalent in Kuwait. Frequent dust storms in the region carry them further into the urban areas, posing an occupational health hazard. The fungal population associated with the respirable (more than 2.5 µm) and inhalable (2.5 µm and less) fractions of aerosols is negligibly explored and warrants comprehensive profiling to pinpoint tAhe health implications. For the present investigation, aerosol was collected using a high-volume air sampler coupled with a six-stage cascade impactor (Tisch Environmental, Inc) at a rate of 566 L min−1. The samples were lysed, DNA was extracted, and the internal transcribed regions were sequenced through targeted amplicon sequencing. Aspergillus, Penicillium, Alternaria, Cladosporium, Fusarium, Gleotinia and Cryptococcus were recorded in all the size fractions with mean relative abundances (RA%) of 17.5%, 12.9%, 12.9%, 4.85%, 4.08%, 2.77%, and 2.51%, respectively. A weak community structure was associated with each size fraction (ANOSIM r2 = 0.11; p > 0.05). The Shannon and Simpson indices also varied among the respirable and inhalable aerosols. About 24 genera were significantly differentially abundant, as described through the Wilcoxon rank sum test (p < 0.05). The fungal microbiome existed as a complex lattice of networks exhibiting both positive and negative correlations and were involved in 428 functions. All the predominant genera were pathogenic, hence, their presence in inhalable fractions raises concerns and poses an occupational exposure risk to both human and non-human biota. Moreover, long-range transport of these fungi to urban locations is undesirable yet plausible. [ABSTRACT FROM AUTHOR] more...

Coastal areas are vital ecosystems, supporting diverse marine life and providing resources essential to human well-being. However, sewage pollution poses a significant threat to these delicate environments, impacting water quality, biodiversity, and public health. Antimicrobial resistance (AMR) has gained importance. We believe the presence of faecal sterols in coastal and marine sediments is a reliable indicator of sewage contamination. At most sites, the faecal contamination was observed with ranges from more...

−1 dw. The coprostanol/cholesterol ratio was >0.2 at 68% of the sampling sites, suggesting a large spatial distribution of sewage contamination beyond the discharge points. The positive correlation of faecal sterols with AMR provides an insight that antibiotic resistance genes (ARGs) can spread to areas away from point sources. We propose that sterol ratios be considered as a screening tool for selecting the sites for AMR assessment. The analyses of sterols will be less time and cost-intensive compared to pharmaceutical analyses and can be a reliable indicator for AMR studies in areas where prior knowledge and experience are lacking. [ABSTRACT FROM AUTHOR]

Objective: Workers in the healthcare sector are exposed to a multitude of bacterial genera. The location of their work contributes significantly to shaping personal microbiomes. In this study, we investigated the role of the workspace on the nasal bacteriome of staff working in a healthcare-associated research facility. Methods: The anterior nares of 10 staff working in different laboratories on the ground and first floor of the research facility were aseptically swabbed. Genomic DNA from each sample was used to amplify the V3 and V4 regions of the 16S rRNA gene. The amplified products were sequenced using the MiSeq sequencer (Illumina). Operational taxonomic units were filtered through MG-RAST v.3.6. Taxonomic profiling and visualizations were performed in MicrobiomeAnalyst v2.0. Results: The Wilcoxson Sum test at median abundances (p < 0.05) indicated that seven taxa (Micromonosporaceae, Micromonospora, Lactobacillaceae, Lactobacillus, Betaproteobacteria, Burkholderiales, Pectobacterium) were significantly diverse between ground-floor and first-floor workers. The analysis of similarity coefficient was 0.412 (p < 0.03) between the ground and the first-floor workers. Random forest analysis predicted 15 features that were significantly different (p < 0.05) in individuals working in different laboratories. Species richness and evenness also differed according to the placement of individuals in respective laboratories. Conclusion: These findings add to the knowledge that the healthcare support staff are at a speculated occupational risk. A slight shift in the abundances of bacterial genera and species might lead to unwanted consequences. Continual monitoring is thus warranted. [ABSTRACT FROM AUTHOR] more...

Kuwaiti territorial waters of the northwest Arabian Gulf represent a unique aquatic ecosystem prone to various environmental and anthropogenic stressors that pose significant constraints on the resident biota which must withstand extreme temperatures, salinity levels, and reducing conditions, among other factors to survive. Such conditions create the ideal environment for investigations into novel functional genetic adaptations of resident organisms. Firstly, however, it is essential to identify said organisms and understand the dynamic nature of their existence. Thus, this study provides the first comprehensive analysis of bacterial and archaeal community structures in the unique waters of Kuwait located in the Northwest Arabian Gulf and analyzes their variations with respect to depth, season, and location, as well as their susceptibility to changes in abundance with respect to various physicochemical parameters. Importantly, this study is the first of its kind to utilize a shotgun metagenomics approach with sequencing performed at an average depth of 15 million paired end reads per sample, which allows for species-level community profiling and sets the framework for future functional genomic investigations. Results showed an approximately even abundance of both archaeal (42.9%) and bacterial (57.1%) communities, but significantly greater diversity among the bacterial population, which predominantly consisted of members of the Proteobacteria, Cyanobacteria, and Bacteroidetes phyla in decreasing order of abundance. Little to no significant variations as assessed by various metrics including alpha and beta diversity analyses were observed in the abundance of archaeal and bacterial populations with respect to depth down the water column. Furthermore, although variations in differential abundance of key genera were detected at each of the three sampling locations, measurements of species richness and evenness revealed negligible variation (ANOVA p<0.05) and only a moderately defined community structure (ANOSIM r2 = 0.243; p>0.001) between the various locations. Interestingly, abundance of archaeal community members showed a significant increase (log2 median ratio of RA = 2.6) while the bacterial population showed a significant decrease (log2 median ratio = -1.29) in the winter season. These findings were supported by alpha and beta diversity analyses as well (ANOSIM r2 = 0.253; p>0.01). Overall, this study provides the first in-depth analysis of both bacterial and archaeal community structures developed using a shotgun metagenomic approach in the waters of the Northwest Arabian Gulf thus providing a framework for future investigations of functional genetic adaptations developed by resident biota attempting to survive in the uniquely extreme conditions to which they are exposed. [ABSTRACT FROM AUTHOR] more...

Microplastics are ubiquitous in aquatic environments, and in most marine environments, copepods are the main metazoans. The ingestion of microplastics by zooplankton is linked to various stresses, including oxidative stress, reduced reproductive capacities, and even mortality in nauplii. Microplastics are also reported to serve as vectors for hydrophobic contaminants. Our experimental results highlight that the retention and contact time of microplastics in copepods is quite short. The experimental results show that Parvocalanus crassirostis and Acartia pacifica defecated 75–84% and 61–71% of ingested microplastics within 60 min of ingestion. The observation raises several questions on the hypothesis of microplastic toxicity and ecological stresses: would a 180-minute contact time result in acute toxicity reported by various workers? An interesting observation was that these two copepod species did not consume microplastics larger than 50 µm in size. Considering this fact, inventories of smaller microplastics might be more important for assessing the ecological effects of MP ingestion among primary consumers in the marine food chain. Another important aspect that this study highlights is the likely change in faecal pellet sinking velocities due to the incorporation of MPs, and faecal pellets are probably efficient vectors for MP transport in the aquatic environment. [ABSTRACT FROM AUTHOR] more...

Prolonged and excessive use of antibiotics has resulted in the development of antimicrobial resistance (AMR), which is considered an emerging global challenge that warrants a deeper understanding of the antibiotic-resistant gene elements (ARGEs/resistomes) involved in its rapid dissemination. Currently, advanced molecular methods such as high-throughput quantitative polymerase chain reaction (HT-qPCR) and shotgun metagenomic sequencing (SMS) are commonly applied for the surveillance and monitoring of AMR in the environment. Although both methods are considered complementary to each other, there are some appreciable differences that we wish to highlight in this communication. We compared both these approaches to map the ARGEs in the coastal sediments of Kuwait. The study area represents an excellent model as it receives recurrent emergency waste and other anthropogenic contaminants. The HT-qPCR identified about 100 ARGs, 5 integrons, and 18 MGEs (total—122). These ARGs coded for resistance against the drug classes of beta-lactams > aminoglycoside > tetracycline, macrolide lincosamide streptogramin B (MLSB) > phenicol > trimethoprim, quinolone, and sulfonamide. The SMS picked a greater number of ARGs (402), plasmid sequences (1567), and integrons (168). Based on the evidence, we feel the SMS is a better method to undertake ARG assessment to fulfil the WHO mandate of "One Health Approach." This manuscript is a useful resource for environmental scientists involved in AMR monitoring. [ABSTRACT FROM AUTHOR] more...

Antibiotic-resistance gene elements (ARGEs) such as antibiotic-resistance genes (ARGs), integrons, and plasmids are key to the spread of antimicrobial resistance (AMR) in marine environments. Kuwait's marine area is vulnerable to sewage contaminants introduced by numerous storm outlets and indiscriminate waste disposal near recreational beaches. Therefore, it has become a significant public health issue and warrants immediate investigation. Coliforms, especially Gram-negative Escherichia coli, have been regarded as significant indicators of recent fecal pollution and carriers of ARGEs. In this study, we applied a genome-based approach to identify ARGs' prevalence in E. coli isolated from mollusks and coastal water samples collected in a previous study. In addition, we investigated the plasmids and intl1 (class 1 integron) genes coupled with the ARGs, mediating their spread within the Kuwait marine area. Whole-genome sequencing (WGS) identified genes resistant to the drug classes of beta-lactams (blaCMY-150, blaCMY-42, blaCTX-M-15, blaDHA-1, blaMIR-1, blaOKP-B-15, blaOXA-1, blaOXA-48, blaTEM-1B, blaTEM-35), trimethoprim (dfrA14, dfrA15, dfrA16, dfrA1, dfrA5, dfrA7), fluroquinolone (oqxA, oqxB, qnrB38, qnrB4, qnrS1), aminoglycoside (aadA2, ant(3")-Ia, aph(3")-Ib, aph(3')-Ia, aph(6)-Id), fosfomycin (fosA7, fosA_6, fosA, fosB1), sulfonamide (sul1, sul2, sul3), tetracycline (tet-A, tet-B), and macrolide (mph-A). The MFS-type drug efflux gene mdf-A is also quite common in E. coli isolates (80%). The plasmid ColRNAI was also found to be prevalent in E. coli. The integron gene intI1 and gene cassettes (GC) were reported to be in 36% and 33%, respectively, of total E. coli isolates. A positive and significant (p < 0.001) correlation was observed between phenotypic AMR-intl1 (r = 0.311) and phenotypic AMR-GC (r = 0.188). These findings are useful for the surveillance of horizontal gene transfer of AMR in the marine environments of Kuwait. [ABSTRACT FROM AUTHOR] more...

Simple Summary: Climate change has negatively affected marine organisms. This experimental study presents data from a multigenerational experiment on the egg production of two commonly occurring calanoid copepods, Parvocalanus crassirostis and Acartia pacifica, under different pH and temperature conditions. The results suggest that pH and temperature conditions influence the number of eggs produced by healthy copepod pairs. However, when the pH changes were gradually carried out over 14 generations, there was no significant difference in the egg production rate at ambient and lower pH. This suggests that copepod populations might be resilient to future ocean scenarios of lower pH and higher temperature. The increasing atmospheric CO2 concentrations and warming of marine waters have encouraged experiments on multi-stressor interactions in marine organisms. We conducted a multigenerational experiment to assess reproductive capacities regarding egg production in calanoid copepods Parvocalanus crassirostis and Acartia pacifica under different pH and temperature conditions. The experimental set-up allowed assessing the tandem effect of warming and acidification on the number of eggs produced by healthy copepod pairs under two pH conditions of 8.20 and 7.50 (hard selection) as well as with a gradual reduction of 0.05 pH units at each generation (soft selection). The results are quite interesting, with very diverse performance across temperatures. The number of eggs produced under hard selection was higher at pH 8.20 compared to pH 7.50 for both species, with the maximum number of eggs produced at 24–28 °C, whereas under soft selection, there was no significant difference in the egg production rate at 24–28 °C across generations and there was an improvement in the number of eggs produced at 8–16 °C. The results provide evidence that in a future ocean scenario of lower pH and higher temperature, the two species, and possibly the copepod population at large, might not decrease. Copepod populations might be resilient, and the transcriptomic evidence of adaptation to increased temperature and lower pH is a ray of hope. We believe further studies are needed to provide more robust datasets to underpin the hypothesis of adaptation to climate change. [ABSTRACT FROM AUTHOR] more...

Antimicrobial resistance (AMR) is one of the biggest threats to human health worldwide. The World Health Organization (WHO, Geneva, Switzerland) has launched the "One-Health" approach, which encourages assessment of antibiotic-resistant genes (ARGs) within environments shared by human-animals-plants-microbes to constrain and alleviate the development of AMR. Aerosols as a medium to disseminate ARGs, have received minimal attention. In the present study, we investigated the distribution and abundance of ARGs in indoor and outdoor aerosols collected from an urban location in Kuwait and the interior of three hospitals. The high throughput quantitative polymerase chain reaction (HT-qPCR) approach was used for this purpose. The results demonstrate the presence of aminoglycoside, beta-lactam, fluoroquinolone, tetracycline, macrolide-lincosamide-streptogramin B (MLSB), multidrug-resistant (MDR) and vancomycin-resistant genes in the aerosols. The most dominant drug class was beta-lactam and the genes were IMP-2-group (0.85), Per-2 group (0.65), OXA-54 (0.57), QnrS (0.50) and OXA-55 (0.55) in the urban non-clinical settings. The indoor aerosols possessed a richer diversity (Observed, Chao1, Shannon's and Pielou's evenness) of ARGs compared to the outdoors. Seasonal variations (autumn vs. winter) in relative abundances and types of ARGs were also recorded (R2 of 0.132 at p < 0.08). The presence of ARGs was found in both the inhalable (2.1 µm, 1.1 µm, 0.7 µm and < 0.3 µm) and respirable (>9.0 µm, 5.8 µm, 4.7 µm and 3.3 µm) size fractions within hospital aerosols. All the ARGs are of pathogenic bacterial origin and are hosted by pathogenic forms. The findings present baseline data and underpin the need for detailed investigations looking at aerosol as a vehicle for ARG dissemination among human and non-human terrestrial biota. [ABSTRACT FROM AUTHOR] more...

The Middle East is a hot spot of dust, and there are reports of as much as 1400 µg m−3 of dust in aerosols from Kuwait, which is among some of the highest dust loadings globally. A significant volume of literature has emerged on dust–air-quality–human-health, and the World Health Organization in its recent air quality guidelines has lowered the limit of annual PM2.5 exposure to 5 μg m−3 from the previous limit of 10 μg m−3. We present a mini-review based on a screening and search of the published data generated in Kuwait on contaminants associated with dust in different size fractions. We also include an unpublished study on organic contaminants in size-fractionated aerosols. The ΣPAHs concentrations in all the six size fractions range between 570 and 3350 pg m−3. The ∑PBDE concentration ranges from ~2 to 1307 pg m−3. The average 210Po activity in aerosol size classes varies between 2289 and 2581 Bq kg−1. The average 210Pb concentration varies between 352 and 412 Bq kg−1. The MP inventory in Kuwait's outdoor aerosol is between 5 and 35 MP in 815 ± 5 m3 of air. The bacterial load in outdoor aerosols is between 6.05 × 103 cells m−3 and 1.24 × 108 cells m−3. The fungal load ranges between 2.11 × 102 cells m−3 and 2.66 × 106 cells m−3. The data suggest that the inhalable fraction of <2.5 µm size contains high concentrations of Polycyclic Aromatic Hydrocarbons (PAHs), Polybrominated diphenyl ethers (PBDEs), 210Po, 210Pb, microplastics, and microbes. These enriched ultrafine aerosols pose a significant risk to human health. The review also highlights the scarcity of contaminant data in respirable and inhalable size fractions that are critical for a comprehensive inhalation risk assessment. [ABSTRACT FROM AUTHOR] more...

Acacia pachyceras O. Schwartz (Leguminoseae), a woody tree growing in Kuwait is critically endangered. High throughput genomic research is immediately needed to formulate effective conservation strategies for its rehabilitation. We therefore, performed a genome survey analysis of the species. Whole genome sequencing generated ~97 Gb of raw reads (92x coverage) with a per base quality score above Q30. The k-mer analysis (17 mer) revealed its genome to be 720Mb in size with an average guanine-cytosine (GC) ratio of 35%. The assembled genome was analyzed for repeat regions (45.4%-interspersed repeats; 9%-retroelements; 2%-DNA transposons). BUSCO assessment of completeness of genome identified 93% of assembly to be complete. Gene alignments in BRAKER2 yielded 34,374 transcripts corresponding to 33,650 genes. Average length of coding sequences and protein sequences were recorded as 1,027nts and 342aa, respectively. GMATA software filtered a total of 901,755 simple sequence repeats (SSRs) regions against which 11,181 unique primers were designed. A subset of 110 SSR primers were PCR validated and demonstrated for its application in genetic diversity analysis of Acacia. The SSR primers successfully amplified A. gerrardii seedlings DNA depicting cross transferability among species. The principal coordinate analysis and the split decomposition tree (bootstrapping runs of 1000 replicates) distributed the Acacia genotypes into two clusters. The flow cytometry analysis revealed the A. pachyceras genome to be polyploid (6x). The DNA content was predicted as 2.46 pg, 1.23 pg, and 0.41 pg corresponding to 2C DNA, 1C DNA and 1Cx DNA, respectively. The results provide a base for further high throughput genomic studies and molecular breeding for its conservation. [ABSTRACT FROM AUTHOR] more...

Coastal sediments in the proximity of wastewater and emergency outfalls are often sinks of pharmaceutical compounds and other organic and inorganic contaminants that are likely to affect the microbial community. The metabolites of these contaminants affect microbial diversity and their metabolic processes, resulting in undesirable effects on ecosystem functioning, thus necessitating the need to understand their composition and functions. In the present investigation, we studied the metagenomes of 12 coastal surface sediments through whole genome shot-gun sequencing. Taxonomic binning of the genes predicted about 86% as bacteria, 1% as archaea, >0.001% as viruses and Eukaryota, and 12% as other communities. The dominant bacterial, archaeal, and fungal genera were Woeseia, Nitrosopumilus, and Rhizophagus, respectively. The most prevalent viral families were Myoviridae and Siphoviridae, and the T4 virus was the most dominant bacteriophage. The unigenes further aligned to 26 clusters of orthologous genes (COGs) and five carbohydrate-active enzymes (CAZy) classes. Glycoside hydrolases (GH) and glycoside transferase (GT) were the highest-recorded CAzymes. The Kyoto Encyclopedia of Genes and Genomes (KEGG) level 3 functions were subjugated by purine metabolism > ABC transporters > oxidative phosphorylation > two-component system > pyrimidine metabolism > pyruvate metabolism > quorum sensing > carbon fixation pathways > ribosomes > and glyoxalate and dicarboxylate metabolism. Sequences allying with plasmids, integrons, insertion sequences and antibiotic-resistance genes were also observed. Both the taxonomies and functional abundances exhibited variation in relative abundances, with limited spatial variability (ANOVA p > 0.05; ANOSIM-0.05, p > 0.05). This study underlines the dominant microbial communities and functional genes in the marine sediments of Kuwait as a baseline for future biomonitoring programs. [ABSTRACT FROM AUTHOR] more...

The wastewater stream is the most significant contributor of microplastics (MPs) to the environment. There are five wastewater treatment plants (WWTPs) in Kuwait. This baseline study provides an overview of MP removal in three major WWTPs in Kuwait that treat some 81.31% of the wastewater produced. The Sulabiya WWTP was the most efficient in MP removal, followed by the Kabd and Umm Al-Haiman WWTPs. The MP removal efficiency of plants in Kuwait is very high for Sulabiya WWTP and Kabd WWTP with an average of 2.5 MP L−1 in treated effluent comparable to the WWTPs in Australia, the United States, and Europe. The standard methodology of sample collection, preparation, and identification using microscopic examination and micro-Raman spectrometry was followed. Over 94.5 billion MPs enter the three WWTPs daily; 92.3 billion MPs are retained in sludge, while 2.2 billion are passed into the environment due to the use of treated effluent. The influent, effluent, and sludge MP inventories ranged between 119 and 230 MP L−1, 1 and 12 MP L−1, and 72 and 103 MP 10 g−1 respectively. The fiber was the dominant shape, and white, transparent, and black were prevalent colors. Currently, sludge is not used in Kuwait for any terrestrial or agricultural application; however, sludge is routinely used in many countries as a soil additive in agricultural farms. Using effluent water in irrigation leads to MP dissemination in the terrestrial environment. It is necessary to assess how far these MPs move in the soil profile and if they can contaminate the shallow aquifers. The observation of MP retention in sludge and effluent is empirical, and the use of these matrixes in agriculture is likely to raise an issue of food safety. [ABSTRACT FROM AUTHOR] more...

The airborne transmission of COVID-19 has drawn immense attention to bioaerosols. The topic is highly relevant in the indoor hospital environment where vulnerable patients are treated and healthcare workers are exposed to various pathogenic and non-pathogenic microbes. Knowledge of the microbial communities in such settings will enable precautionary measures to prevent any hospital-mediated outbreak and better assess occupational exposure of the healthcare workers. This study presents a baseline of the bacterial and fungal population of two major hospitals in Kuwait dealing with COVID patients, and in a non-hospital setting through targeted amplicon sequencing. The predominant bacteria of bioaerosols were Variovorax (9.44%), Parvibaculum (8.27%), Pseudonocardia (8.04%), Taonella (5.74%), Arthrospira (4.58%), Comamonas (3.84%), Methylibium (3.13%), Sphingobium (4.46%), Zoogloea (2.20%), and Sphingopyxis (2.56%). ESKAPEE pathogens, such as Pseudomonas, Acinetobacter, Staphylococcus, Enterococcus, and Escherichia, were also found in lower abundances. The fungi were represented by Wilcoxinia rehmii (64.38%), Aspergillus ruber (9.11%), Penicillium desertorum (3.89%), Leptobacillium leptobactrum (3.20%), Humicola grisea (2.99%), Ganoderma sichuanense (1.42%), Malassezia restricta (0.74%), Heterophoma sylvatica (0.49%), Fusarium proliferatum (0.46%), and Saccharomyces cerevisiae (0.23%). Some common and unique operational taxonomic units (OTUs) of bacteria and fungi were also recorded at each site; this inter-site variability shows that exhaled air can be a source of this variation. The alpha-diversity indices suggested variance in species richness and abundance in hospitals than in non-hospital sites. The community structure of bacteria varied spatially (ANOSIM r2 = 0.181–0.243; p < 0.05) between the hospital and non-hospital sites, whereas fungi were more or less homogenous. Key taxa specific to the hospitals were Defluvicoccales, fungi, Ganodermataceae, Heterophoma, and H. sylvatica compared to Actinobacteria, Leptobacillium, L. leptobacillium, and Cordycipitaceae at the non-hospital site (LefSe, FDR q ≤ 0.05). The hospital/non-hospital MD index > 1 indicated shifts in the microbial communities of indoor air in hospitals. These findings highlight the need for regular surveillance of indoor hospital environments to prevent future outbreaks. [ABSTRACT FROM AUTHOR] more...

Marine sediments are a sink for antibiotic resistance genes (ARGs) and antibiotic-resistant microbes (ARMs). Wastewater discharge into the aquatic environment is the dominant pathway for pharmaceuticals reaching aquatic organisms. Hence, the characterization of ARGs is a priority research area. This baseline study reports the presence of ARGs in 12 coastal sediment samples covering the urban coastline of Kuwait through whole-genome metagenomic sequencing. The presence of 402 antibiotic resistance genes (ARGs) were recorded in these samples; the most prevalent were patA, adeF, ErmE, ErmF, TaeA, tetX, mphD, bcrC, srmB, mtrD, baeS, Erm30, vanTE, VIM-7, AcrF, ANT4-1a, tet33, adeB, efmA, and rpsL, which showed resistance against 34 drug classes. Maximum resistance was detected against the beta-lactams (cephalosporins and penam), and 46% of genes originated from the phylum Proteobacteria. Low abundances of ESKAPEE pathogens (Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumonia, Acinetobacter baumanii, Pseudomonas aeruginosa, Enterobacter sps., and Escherichia coli) were also recorded. Approximately 42% of ARGs exhibited multiple drug resistance. All the ARGs exhibited spatial variations. The major mode of action was antibiotic efflux, followed by antibiotic inactivation, antibiotic target alteration, antibiotic target protection, and antibiotic target replacement. Our findings supported the occurrence of ARGs in coastal marine sediments and the possibility of their dissemination to surrounding ecosystems. [ABSTRACT FROM AUTHOR] more...

Natural populations of Rhanterium eppaposum Oliv. (Arfaj), a perennial forage shrub, have depleted due to unethical human interventions and climate change in Kuwait. Therefore, there is an urgent need to conserve this native plant through the assessment of its genetic diversity and population structure. Genotyping by sequencing (GBS) has recently emerged as a powerful tool for the molecular diversity analysis of higher plants without prior knowledge of their genome. This study represents the first effort in using GBS to discover genome-wide single nucleotide polymorphisms (SNPs) of local Rhanterium plants to assess the genetic diversity present in landraces collected from six different locations in Kuwait. The study generated a novel set of 11,231 single nucleotide polymorphisms (SNPs) and indels (insertions and deletions) in 98 genotypes of Rhanterium. The analysis of molecular variance (AMOVA) revealed ~1.5% variation residing among the six populations, ~5% among the individuals within the population and 93% variation present within the populations (FST = 0.029; p = 0.0). Bayesian and UPGMA analyses identified two admixed clusters of the tested samples; however, the principal coordinates analysis returned the complete population as a single group. Mantel's test returned a very weak correlation coefficient of r2 = 0.101 (p = 0.00) between the geographic and genetic distance. These findings are useful for the native species to formulate conservation strategies for its sustainable management and desert rehabilitation. [ABSTRACT FROM AUTHOR] more...

This study provides baseline information on the concentrations of antibiotics in influent and effluent from two wastewater treatment plants in regular operation in the State of Kuwait. Wastewater samples were collected from the influent and effluent streams of two WWTPs, over four sampling campaigns and analyzed for a broad range of antibiotics. The mean influent concentrations of sulfamethoxazole, ciprofloxacin, clarithromycin, and cefalexin were 852 ng/L, 672 ng/L, 592 ng/L), and 491 ng/L, respectively, at Umm Al Hayman WWTP. At the Kabd WWTP, the influent concentration of clarithromycin was highest with a mean of 949 ng/L, followed by ciprofloxacin (mean, 865 ng/L), cefalexin (mean, 598 ng/L), and sulfamethoxazole (mean, 520 ng/L). The dominant compounds in the effluent from Umm Al Hayman were sulfamethoxazole (mean, 212 ng/L), ciprofloxacin (mean, 153 ng/L), ofloxacin (mean, 120 ng/L), dimetridazole (mean, 96 ng/L), and metronidazole (mean, 93 ng/L). Whereas, at the Kabd WWTP, the dominant compounds were sulfamethoxazole (mean, 338 ng/L), dimetridazole (mean, 274 ng/L), cefalexin (mean, 213 ng/L), ciprofloxacin (mean, 192 ng/L), and clarithromycin (189 ng/L). The mean influent concentrations of all compounds were higher than those measured in the effluents. The concentrations of antibiotic compounds were not significantly different between the two WWTPs (p > 0.05). The removal efficiencies of the various antibiotics over the four sampling campaigns for the Kabd and Umm Hayman WWTPs ranged between 10.87 and 99.75% and also showed that they were variable and were compound dependent. The data clearly show that the concentrations of antibiotics measured in the influents of both WWTPs were highest in samples collected during the winter-summer (September samples) transition followed by the concentrations measured during the winter-summer (March samples) transition period. This is possibly linked to the increased prescription of these medications to treat infectious diseases and flu prevalent in Kuwait during these periods. This study provides the first reported concentrations of antibiotics in the dissolved aqueous influents and effluents of WWTPs in Kuwait. Additional studies are required to evaluate the environmental impact that antibiotic residues may cause since treated wastewater is used in irrigation, and often there are instances when untreated wastewater is discharged directly into the marine environment. [ABSTRACT FROM AUTHOR] more...

The microorganisms at the workplace contribute towards a large portion of the biodiversity a person encounters in his or her life. Health care professionals are often at risk due to their frontline nature of work. Competition and cooperation between nasal bacterial communities of individuals working in a health care setting have been shown to mediate pathogenic microbes. Therefore, we investigated the nasal bacterial community of 47 healthy individuals working in a clinical research laboratory in Kuwait. The taxonomic profiling and core microbiome analysis identified three pre-dominant genera as Corynebacterium (15.0%), Staphylococcus (10.3%) and, Moraxella (10.0%). All the bacterial genera exhibited seasonal variations in summer, winter, autumn and spring. SparCC correlation network analysis revealed positive and negative correlations among the classified genera. A rich set of 16 genera (q < 0.05) were significantly differentially abundant (LEfSe) across the four seasons. The highest species counts, richness and evenness (P < 0.005) were recorded in autumn. Community structure profiling indicated that the entire bacterial population followed a seasonal distribution (R2-0.371; P < 0.001). Other demographic factors such as age, gender and, ethnicity contributed minimally towards community clustering in a closed indoor laboratory setting. Intra-personal diversity also witnessed rich species variety (maximum 6.8 folds). Seasonal changes in the indoor working place in conjunction with the outdoor atmosphere seems to be important for the variations in the nasal bacterial communities of professionals working in a health care setting. [ABSTRACT FROM AUTHOR] more...

Kuwait is a country with a very high dust loading; in fact it bears the world's highest particulate matter concentration in the outdoor air. The airborne dust often has associated biological materials, including pathogenic microbes that pose a serious risk to the urban ecosystem and public health. This study has established the baseline taxonomic characterization of microbes associated with dust transported into Kuwait from different trajectories. A high volume air sampler with six-stage cascade impactor was deployed for sample collection at a remote as well as an urban site. Samples from three different seasons (autumn, spring and summer) were subjected to targeted amplicon sequencing. A set of ~ 50 and 60 bacterial and fungal genera, respectively, established the core air microbiome. The predominant bacterial genera (relative abundance ≥ 1%) were Brevundimonas (12.5%), Sphingobium (3.3%), Sphingopyxis (2.7%), Pseudomonas (2.5%), Sphingomonas (2.4%), Massilia (2.3%), Acidovorax (2.0%), Allorhizobium (1.8%), Halomonas (1.3%), and Mesorhizobium (1.1%), and the fungal taxa were Cryptococcus (12%) followed by Alternaria (9%), Aspergillus (7%), Candida (3%), Cladosporium (2.9%), Schizophyllum (1.6%), Fusarium (1.4%), Gleotinia (1.3%) and Penicillium (1.15%). Significant spatio-temporal variations were recorded in terms of relative abundances, α-diversities, and β-diversities of bacterial communities. The dissimilarities were less pronounced and instead the communities were fairly homogenous. Linear discrimant analysis revealed three fungal genera known to be significantly differentially abundant with respect to different size fractions of dust. Our results shed light on the spatio-temporal distribution of airborne microbes and their implications in general health. [ABSTRACT FROM AUTHOR] more...

Objective: The identification of Brucella genotypes is essential for epidemiological studies. The whole-genome sequencing is emerging as a novel tool for genetic characterization of infectious microbes. The aim of this study was to genotype Brucella melitensis isolates from Kuwait using whole-genome sequencing and variant analysis of the sequence data. Methods: DNA was purified from 15 heat-inactivated B. melitensis isolates and used to prepare sequencing libraries employing Nextera XT DNA Sample Preparation Kit (Illumina San Diego, CA, USA) and sequenced on a MiSeq (Illumina). The sequence files were aligned to three biovars of B. melitensis, i.e., biovar 1 str. 16M, biovar 2 str. 63/9, and biovar 3 str. Ether. The alignment and variant calling were performed using “bwa-mem” and SAMtools/VCFtools, respectively. Results: The genome size of all the isolates was around 3.3 mega base pairs and resembled B. melitensis biovar 2. Single-nucleotide polymorphisms (SNPs), insertions, and deletions (indels) were spread all over the genome; but 138 SNPs were common among the 14 isolates, supporting the same ancestral origin. A neighbor-joining tree analysis identified isolate 2 as an outlier. In addition, SNPs (2–478) specific to each isolate were also identified, which divided the B. melitensis biovar 2 into two major groups/genotypes. A further analysis showed that the Kuwaiti isolates of the present study shared phylogeny mainly with strains from the Middle Eastern countries. Conclusions: Among the 15 studied isolates from Kuwait, biovar 2 is the most prevalent biovar of B. melitensis. Furthermore, isolate-specific genetic variations were identified, which may be useful in epidemiological investigations. [ABSTRACT FROM AUTHOR] more...

Haloxylon salicornicum moq. Bunge ex Boiss (Rimth) is one of the native plants of Kuwait, extensively depleting through the anthropogenic activities. It is important to conserve Haloxylon community in Kuwait as it can tolerate extreme adverse conditions of drought and salinity to be potentially used in the desert and urban revegetation and greenery national programs. Therefore, a set of 16 inter simple sequence repeat (ISSR) markers were used to assess genetic diversity and population structure of 108 genotypes from six locations in Kuwait. The ISSR primers produced 195 unambiguous and reproducible bands out of which 167 bands were polymorphic (86.5%) with a mean PIC value of 0.31. The overall average values of Nei’s gene diversity (h') and Shannon’s diversity index (I) were 0.254 and 0.375, respectively. Results of AMOVA revealed high genetic variations within populations (77.8%) and low among populations (22%). The values of Fixation index (FST = 0.22; P = 0.0), Genetic differentiation (GST = 0.262; G’ST = 0.327; D = 0.335 and Gene flow (NM = 0.880) were indicative of heterozygous populations. The results of STRUCTURE and split decomposition analysis suggested that the Rimth accessions of Kuwait can be grouped into five and six subpopulations, respectively. Principal coordinate analysis (PCoA) grouped them into three clusters. The pairwise Nei’s genetic distances (DS) among populations demonstrated a narrow range from 0.047 to 0.187 (Scale-0.0 to 1.0). The Mantel’s test revealed a weak correlation (r2- 0.188; P-0.013) between the genetic distance and geographic distances. Our results suggest that the narrowly distributed Haloxylon community in Kuwait demonstrated a high genetic diversity within the populations however the overall population structure was weak. [ABSTRACT FROM AUTHOR] more...

The correct Funding statement is: The study was funded by Kuwait University Research Sector's grants RM01/13 and SRUL02/13. Reference 1 Habibi N, Mustafa AS, Khan MW (2021) Composition of nasal bacterial community and its seasonal variation in health care workers stationed in a clinical research laboratory. [Extracted from the article] more...

Brucellosis is a zoonotic disease of major concern in Kuwait and the Middle East. Human brucellosis can be caused by several Brucella species with varying degree of pathogenesis, and relapses are common after apparently successful therapy. The classical biochemical methods for identification of Brucella are time-consuming, cumbersome, and provide information limited to the species level only. In contrast, molecular methods are rapid and provide differentiation at intra-species level. In this study, four molecular methods [16S rRNA gene sequencing, real-time PCR, enterobacterial repetitive intergenic consensus (ERIC)-PCR and multilocus variable-number tandem-repeat analysis (MLVA)-8, MLVA-11 and MLVA-16 were evaluated for the identification and typing of 75 strains of Brucella isolated in Kuwait. 16S rRNA gene sequencing of all isolates showed 90–99% sequence identity with B. melitensis and real-time PCR with genus- and species- specific primers identified all isolates as B. melitensis. The results of ERIC-PCR suggested the existence of 75 ERIC genotypes of B. melitensis with a discriminatory index of 0.997. Cluster classification of these genotypes divided them into two clusters, A and B, diverging at ~25%. The maximum number of genotypes (n = 51) were found in cluster B5. MLVA-8 analysis identified all isolates as B. melitensis, and MLVA-8, MLVA-11 and MLVA-16 typing divided the isolates into 10, 32 and 71 MLVA types, respectively. Furthermore, the combined minimum spanning tree analysis demonstrated that, compared to MLVA types discovered all over the world, the Kuwaiti isolates were a distinct group of MLVA-11 and MLVA-16 types in the East Mediterranean Region. [ABSTRACT FROM AUTHOR] more...

Simple Summary: The topic of plastic wastes, microplastic (MP) and nanoplastic (NP) particles in the aquatic environment has been the focus of much scientific effort over the last decade and has gained immense public attention through the media. While numerous scientific reports underscore the ubiquitous presence of MPs and NPs in aquatic environments, particularly the oceans, there are many unresolved issues involved in their sampling, identification and characterization. This paper addresses some of the main problems and suggests what needs to be undertaken to overcome these issues. An overriding problem is the lack of harmonization of the protocols used for sampling MPs at sea and identifying them in the laboratory. There are technological challenges in polymeric characterization of NPs in environmental samples. Researchers use a wide variety of net types and net mesh sizes to capture and separate MP floating in the sea which makes comparing MP concentrations from different teams and areas extremely difficult and calls for establishing inter-comparison exercises among the various research teams. Furthermore, the issue of whether chemicals in MPs and NPs leach following ingestion by biota or whether they transport and release contaminants adsorbed on MP/NP surfaces is still unresolved. In essence, these and other issues have to be addressed and resolved before society has an accurate picture of their importance as an aquatic pollutant. There are no datasets on the environmental concentration of NPs, hence their effect on biota is solely relied on laboratory experiments using extremely high concentrations. The legitimacy of these effects and interactions in the biotic system is something that warrants discussion. Microplastic research has become a buzz word. It is seen as one of the most pressing issues of Anthropocene contamination. There is certainly no doubt about the ubiquitous presence of microplastic (MP) in almost all environmental matrices. However, the validity of considering them as a vector for contaminants needs some reconsideration, there are other more potent pathways. Their effect on marine biota also calls for some realistic experiments with environmental concentrations of MP and nanoplastic (NP). It has been observed that in most published literature, polymer characterization is performed. Is it necessary to do, or will merely finding and confirming the particle as plastic suffice for environmental research? Harmonization of protocols is necessary, and there is likely a need for some inter-laboratory comparison exercises in order to produce comparable data and reliable assessments across regions. Samples collected from the same area using different techniques show an order of magnitude difference in MP concentration. The issue of nanoplastic is more contentious; are we technologically ready to identify NP in environmental samples? [ABSTRACT FROM AUTHOR] more...

The omnipresence of microplastic (MP) in various environmental samples, including aerosols, has raised public health concerns; however, there is presently very limited information on MPs in indoor aerosol. This paper presents a unique dataset where smaller MPs have been sampled using a six-stage cascade impactor from indoor environments in Kuwait. The MP concentration in the indoor air varied between 3.2 and 27.1 particles m−3, and the relative MP concentration decreased linearly from the lowest to the highest size fraction. A significant effect of location was observed for the total number of MPs (F2,14 = 5.80, p = 0.02) and the inhalable fraction (F2,14 = 8.38, p = 0.005), while location had no effect on the respirable fraction (F2,14 = 0.54, p = 0.60). A significant effect of the type of air conditioning used was also observed for the total number of MPs (F2,19 = 5.58, p = 0.01) and the inhalable fraction (F2,19 = 6.45, p = 0.008), while location had no effect on the respirable fraction (F2,19 = 1.30, p = 0.30). For the total number of MPs and the inhalable fraction, the concentration was significantly higher for the split unit air-conditioning as compared to the central air-conditioning plants. The presence/absence of carpets had no significant effect on the MP concentrations (total: F1,19 = 4.08, p = 0.06; inhalable: F1,19 = 3.03, p = 0.10; respirable: F1,19 = 4.27, p = 0.05). The shape was dominantly fibers, with few fragments in lower size fractions. These datasets represent the first baseline information for Kuwait, and the smaller MPs in all the samples further underscore the need to develop standardized protocols of MP collection in the ≤2.5 µm fraction that can have more conspicuous health implications. [ABSTRACT FROM AUTHOR] more...

The dinoflagellates of the genus Gambierdiscus are found in almost all oceans and seas between the coordinates 35° N and 35° S. Gambierdiscus and Fukuyoa are producers of ciguatoxins (CTXs), which are known to cause foodborne disease associated with contaminated seafood. The occurrence and effects of CTXs are well described in the Pacific and the Caribbean. However, historically, their properties and presence have been poorly documented in the Indian Ocean (including the Bay of Bengal, Andaman Sea, and the Gulf). A higher occurrence of these microorganisms will proportionately increase the likelihood of CTXs entering the food chain, posing a severe threat to human seafood consumers. Therefore, comprehensive research strategies are critically important for developing effective monitoring and risk assessments of this emerging threat in the Indian Ocean. This review presents the available literature on ciguatera occurrence in the region and its adjacent marginal waters: aiming to identify the data gaps and vectors. [ABSTRACT FROM AUTHOR] more...

Acacia tree population is declining in several countries of the world especially in the Arabian peninsula due to human-induced activities. The tree has potential medicinal and economic benefits as a source of fuel and timber. It can fix nitrogen, a significant property that assists in desert rehabilitation. However, the lack of genomic information of Acacia pachyceras hampers its genetic study and breeding process. We performed paired-end sequencing of A. pachyceras at a depth of 120X to obtain raw sequences of 108.9 GB with a per base quality >Q30. Filtered raw data was assembled into a fasta file of 4 GB. The assembled genomic sequences consisted of 901,755 single sequence repeats (SSRs). In total 11,596 primer pairs were designed against these SSR motifs. The data generated provides baseline genomic information about the species and formulates a base for further sequencing of A. pachyceras through PACBio and HiC technologies. The novel developed SSR markers will facilitate genetic diversity and conservation studies for Acacia species., 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., (© 2022 The Author(s). Published by Elsevier Inc.) more...

Human brucellosis is a neglected and underrecognized infection of widespread geographic distribution. Brucellosis is present on all inhabited continents and endemic in many areas of the world, including Kuwait and the Middle East. Here, we present draft genome assemblies of five Brucella melitensis strains isolated from brucellosis patients in Kuwait., (Copyright © 2016 Khan et al.) more...