Spatial Variations in the Nasal Microbiota of Staff Working in a Healthcare-Associated Research Core Facility.

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.

Diversity and spatiotemporal variations in bacterial and archaeal communities within Kuwaiti territorial waters of the Northwest Arabian Gulf.

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.

Correction: Composition of nasal bacterial community and its seasonal variation in health care workers stationed in a clinical research laboratory.

[This corrects the article DOI: 10.1371/journal.pone.0260314.].

Antibiotic Resistance Mediated by Escherichia coli in Kuwait Marine Environment as Revealed through Genomic Analysis.

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.

Data on SSR markers and SNPs filtered from transcriptome of Parvocalanus crassirostris.

Calanoid copepod populations are being severely affected due to the effects of ocean acidification (OA) and ocean warming (OW). These marine organisms are the most abundant primary consumers contributing significantly in the marine food web. Any effect on the abundance and diversity of copepods due to climate change is likely to have serious implications on the marine ecosystem functioning. Molecular studies that play a vital role in assessing the genetic changes under the influence of environmental imbalances are completely lacking for this species. Here we report the genetic variations in three generations of copepods through transcriptome sequencing. RNA sequencing was performed on an Illumina HiSeq platform employing the 2 × 100 bp paired-end chemistry. Approximately, 10GB of data was obtained for all the samples. The raw sequences were assembled through Trinity 2.6.6 and mined for single nucleotide polymorphisms (SNPs) and simple sequence repeats (SSRs). MIcroSAtellite Identification Tool (MISA) was used for SSR detection and Primer 3 (v 3.0) was utilized to design short oligonucleotide primers (18-20 mers). A total of 15,222 SSRs were identified and 28,944 primer pairs were designed against these motifs. The transcriptome possessed 413,890 SNPs at a frequency of 2.8 per kb. The newly discovered SSRs and SNPs could act as genetic markers for future studies on genetic diversity and conservation for Parvocalanus crassirostris.

The Reproductive Capacities of the Calanoid Copepods Parvocalanus crassirostis and Acartia pacifica under Different pH and Temperature Conditions.

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.

Antibiotic Resistance Genes in Aerosols: Baseline from Kuwait.

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.

Genome survey and genetic characterization of Acacia pachyceras O. Schwartz.

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.

Metagenomes from Coastal Sediments of Kuwait: Insights into the Microbiome, Metabolic Functions and Resistome.

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.

Bacterial and fungal communities in indoor aerosols from two Kuwaiti hospitals.

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 r 2 = 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.

Comparison and optimization of DNA Isolation protocols for high throughput genomic studies of Acacia pachyceras Schwartz.

We describe the optimization and validation of six DNA isolation protocols from fresh leaves of the rare tree Acacia pachyceras. The first four protocols employed three commercial kits (Sigma, Nucleospin1, Nucleospin 2, Promega) whereas the remaining two were based on the traditional sodium dodecyl sulfate (SDS) and cetyltrimethylammonium bromide CTAB methods. Each protocol provided significantly different results concerning DNA concentration (p < 0.032), yield (p < 0.000), contaminant carry over, protocol duration, cost per sample, and comprehensive cost. We demonstrated the applicability of all the tested protocols in DNA barcoding. The protocol yielded maximum amounts (92.85 µg) of DNA in a rapid turnaround time (8 h). The quantity and purity surpassed all the other tested methods. DNA extracted by the CTAB method was the best for NGS (Phred score >Q30). These protocols will be useful tools for molecular research of Acacia pachyceras and other closely related tree species.

Dataset of 16S rRNA and alkB genes in hydrocarbon polluted soils of Kuwait as revealed by Pyrosequencing.

The data in this article was generated by high throughput sequencing of moderately hydrocarbon polluted sites (S1 and S2) and a heavily polluted site (S3) in Kuwait. Deoxyribonucleic acid (DNA) extracted from each site was subjected to polymerase chain reaction (PCR) amplification employing conserved primers of 16S rRNA and alkB genes. Unique Molecular Identifiers (MID) tags were added to individual samples prior to pooling and sequencing on a Roche GS FLX platform using Pyrosequencing Titanium Chemistry. Raw sff files were deposited to the public repository of National Centre for Biotechnology Information (NCBI) under accession no PRJNA816075. The sff files were clipped according to the MID tags and converted to fasta format. 16S rRNA gene sequences were aligned against the SILVA database. The predominant genera at S1 and S2 was Alkanindiges whereas Alcanivorax, was highly abundant at S3. Alkanindiges have been found to play a key role in hydrocarbon degradation and Alcanivorax genus is known for its hydrocarbon degrading capability. The alk B gene sequences were subjected to blastx. The diversity of alkB gene was higher in S3 as compared to S1 and S2. These findings may open the way to the use of the genera Alkanindiges and Alcanivorax in the rehabilitation of hydrocarbon-contaminated sites in hot, arid climates. The isolation of these microorganisms and the design of bioaugmentation procedures specific to the dry climate could be a key step towards the restoration of hydrocarbon contaminated soils.

Genetic Diversity of Rhanterium eppaposum Oliv. Populations in Kuwait as Revealed by GBS.

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.

Antibiotics in Wastewater: Baseline of the Influent and Effluent Streams in Kuwait.

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.

Draft genome sequence and SSR mining data of Acacia pachyceras Schwartz.

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.

A Preliminary Assessment of Size-Fractionated Microplastics in Indoor Aerosol-Kuwait's Baseline.

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.

Micro-Nano Plastic in the Aquatic Environment: Methodological Problems and Challenges.

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?

Data on draft genome assembly and annotation of Haloxylon salicornicum Moq.

Haloxylon salicornicum Moq. Bunge ex Boiss (Rimth) is one of the main structural elements in Eastern Arabian vegetation associations. The plant is utilized as a food source for domestic stock, stabilizes the soil surface besides providing suitable microclimates for exotic species. It is considered one of the most promising species for re-vegetation. H. salicornicum community is under threat from overgrazing leading to a reduction in the percentage of distribution from 22.7% to 2.2% in Kuwait. Therefore, genome characterization of this important Kuwaiti plant is required to formulate strategies for its conservation. Here we report the draft of the H. salicornicum genome, which was sequenced on an Illumina HiSeq 2500 platform. BUSCO assessment revealed 69% of the genome was to be complete. Overall, 12960 gene structures, 11280 protein-coding genes, 11309 mRNAs (protein-coding), 51265 exons and 48100 CDSs were predicted. Functional annotation was carried out by interproscan-5.29-68.0. A total of 7222 protein-coding sequences were, annotated out of 11309 by at least one ontology term. All these genes were associated with 11 major biological processes branched into 60 child processes.

SARS-CoV-2 in hospital air as revealed by comprehensive respiratory viral panel sequencing.

BACKGROUND: Nosocomially acquired severe acute respiratory syndrome coronavirus type 2 (SARS-CoV-2) infection has become the most significant pandemic of our lifetime. Though its transmission was essentially attributed to droplets from an infected person, with recent advancements in knowledge, aerosol transmission seems to be a viable pathway, as well. Because of the lower biological load in ambient aerosol, detection of SARS-CoV-2 is challenging. A few recent attempts of sampling large aerosol volumes and using next-generation sequencing (NGS) to detect the presence of SARS-CoV-2 in the air at very low levels gave positive results. These results suggest the potential of using this technique to detect the presence of SARS-CoV-2 and use it as an early warning signal for possible outbreak or recurrence of coronavirus disease 2019 (COVID-19). AIM: To assess efficacy of comprehensive respiratory viral panel (CRVP) sequencing and RT-PCR for low-level identification of SARS-CoV-2 and other respiratory viruses in indoor air. METHODS: A large volume of indoor aerosol samples from three major hospitals involved in COVID-19 care in Kuwait was collected. Viral RNA was isolated and subjected to comprehensive respiratory viral panel sequencing (CRVP) as per the standard protocol to detect the SARS-CoV-2 and other respiratory viruses in the hospital aerosol and monitor variations within the sequences. RT-PCR was also employed to estimate the viral load of SARS-CoV-2. FINDINGS: 13 of 15 (86.7%) samples exhibited SARS-CoV-2 with a relative abundance of 0.2-33.3%. The co-occurrence of human adenoviruses (type C1, C2, C5, C4), respiratory syncytial virus (RSV), influenza B, and non-SARS-CoV-229E were also recorded. Alignment of SARS-CoV-2 sequences against the reference strain of Wuhan China revealed variations in the form of single nucleotide polymorphisms (SNPs-17), insertions and deletions (indels-1). These variations were predicted to create missense (16), synonymous (15), frameshift (1) and stop-gained (1) mutations with a high (2), low (15), and moderate (16) impact. CONCLUSIONS: Our results suggest that using CRVP on a large volume aerosol sample was a valuable tool for detecting SARS-CoV-2 in indoor aerosols of health care settings. Owing to its higher sensitivity, it can be employed as a surveillance strategy in the post COVID times to act as an early warning system to possibly control future outbreaks.

Enhanced Polonium Concentrations in Aerosols from the Gulf Oil Producing Region and the Role of Microorganisms.

This study provides the first data set of 210Po and 210Pb activity concentrations in the organic and inorganic components of several particle size classes of aerosols collected at two sampling stations in Kuwait. The 210Po concentrations in the aerosols (Bq/g) were similar in all of the particle size classes, but as most (91%) of the aerosol load was made of fine fraction particles of PM0.39-2.5 µm, most of the 210Po activity was carried by this aerosol fraction. At the two sampling stations, the 210Po/210Pb activity concentration ratios in the aerosols were similar, stable around the year, and averaged 1.5 (range 1.2-1.9), much higher than the typical activity concentration ratios of these radionuclides in unmodified (background) aerosols, with Po/Pb < 0.1. The aerosol enrichment in 210Po was likely originated from the oil industry, specifically by gas flaring and oil refining in the Gulf region. Radionuclide analysis in the organic and inorganic components of aerosols showed that the 210Po concentration in the organic component was one order of magnitude higher than the 210Po concentration in the inorganic component, in contrast with 210Pb, which displayed similar concentrations in both organic and inorganic aerosol components. The 210Po carrying organic component of aerosols was investigated and it was found to be largely composed of microorganisms with high microbial and fungi diversity, with the phyla Proteobacteria, Ascomycota, and Basidiomycota being dominant among the bacteria and with Zygomycota being dominant among the fungi. Therefore, we are facing an active concentration process of the atmospheric 210Po carried out by microorganisms, which underlies the 210Po enrichment process in the organic component of aerosols. This bioconcentration of polonium in bioaerosols was unknown.