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The role of airborne particles in the spread of severe acute respiratory syndrome coronavirus type 2 (SARS‐CoV‐2) is well explored. The novel coronavirus can survive in aerosol for extended periods, and its interaction with other viral communities can cause additional virulence and infectivity. This baseline study reports concentrations of SARS‐CoV‐2, other respiratory viruses, and pathogenic bacteria in the indoor air from three major hospitals (Sheikh Jaber, Mubarak Al‐Kabeer, and Al‐Amiri) in Kuwait dealing with coronavirus disease 2019 (COVID‐19) patients. The indoor aerosol samples showed 12–99 copies of SARS‐CoV‐2 per m3 of air. Two non‐SARS‐coronavirus (strain HKU1 and NL63), respiratory syncytial virus (RSV), and human bocavirus, human rhinoviruses, Influenza B (FluB), and human enteroviruses were also detected in COVID‐positive areas of Mubarak Al Kabeer hospital (MKH). Pathogenic bacteria such as Mycoplasma pneumonia, Streptococcus pneumonia and, Haemophilus influenza were also found in the hospital aerosols. Our results suggest that the existing interventions such as social distancing, use of masks, hand hygiene, surface sanitization, and avoidance of crowded indoor spaces are adequate to prevent the spread of SARS‐CoV‐2 in enclosed areas. However, increased ventilation can significantly reduce the concentration of SARS‐CoV‐2 in indoor aerosols. The synergistic or inhibitory effects of other respiratory pathogens in the spread, severity, and complexity of SARS‐CoV‐2 need further investigation.

The role of airborne particles in spread of remains largely unexplored. It has been speculated that the novel corona virus can survive for extended periods in aerosols and its interaction with other viral communities is responsible for additional virulence and infectivity. Therefore, investigations on adsorption, survival, and behavior of the COVID-19 virus within the aerosol community are needed to help understand its spread. In order to explore its spread via aerosols an immediate need is to develop efficient cost-effective sampling methodology for viral aerosols. In view of this we performed the aerosol sample collection through a simplified protocol adapted for its use in laboratory research with minimal biosafety regulations level 1 biosafety level precautions and facilities. In this setup, the air was passed through three gas glass bottles filled with TRIzol @ 30 L−1. The latter served the purpose of collecting and lysing the viral particles trapped in the air. The collected lysate can be transported safely to biosafety regulations level 1 class biosafety level laboratories for downstream processing of ribonucleic acid purification and further analysis such as quantitative polymerase chain reaction or next generation sequencing-based applications. We tested the viability status of the collected aerosols in TRIzol and discovered 90%–100% of the microbial load to be lysed. We expect to recover approximately 1 µg of total ribonucleic acid from 3.6 m3 of aerosols that was successfully amplified using bacterial, fungal, and viral primers. Hence, this technique is safe for use in laboratories that are not complying with the stringent requirements of a virology laboratory.

Kuwait’s wastewater treatment plants experience severe sludge bulking and foaming problems during the winter months. In an effort to identify the possible causes of these problems, sample of wastewater and sludge was collected from Um-Al-Haiman and Riqqa activated sludge systems each week for 32 weeks. The quality of wastewater was determined following standard methods of water analysis, while the dominant filamentous bacteria were identified and quantified using a molecular method called Vermicon Identification Technology (VIT). Multivariate statistical analysis (namely, Pearson’s correlation analysis and principal component analysis) was then carried out on the obtained datasets of results. Pearson’s correlation analysis revealed several significant associations between filamentous bacteria growth and influent quality and operational modes for the systems. The principal component analysis revealed six components that accounted for more than 70% of the total datasets’ information. The techniques used in this study can also be used to assess the relationship between filamentous bacteria growth and the operating conditions of other activated sludge systems.

Airborne particles play a significant role in the spread of bacterial communities. The prevalence of both pathogenic and non-pathogenic forms in the inhalable fractions of aerosols is known. The abundance of microorganisms in the aerosols heightens the likely health hazards due to inhalation since they serve as carriers for pathogens and allergens, often acting as a vector for pulmonary/respiratory infections. Not much information is available on the occurrence and prevalence of bacterial communities in different size-fractionated aerosols in Kuwait. A high-volume air sampler with a six-stage cascade impactor was deployed for sample collection at two sites representing a remote and an urban site. A total volume of 815 ± 5 m3 of air was passed through the filters to trap the particulate matter ranging from 0.39 to >10.2 μm in size (Stage 1 to Stage 5 and base filter). Aeromonas dominated all the stages at the urban site and Stage 5 at the remote site, whereas Sphingobium was prevalent at Stages, 2, 3 and 4 at the remote site. Brevundimonas were found at Stages 1 and 5, and the base filter at the remote site. These results show that the bacterial community is altered in different size fractions of aerosols. Stages 1–4 form the respirable fraction, whereas Stage 5 and particles on the base filter are the inhalable fractions. Many species of Aeromonas cause disease, and hence their presence in inhalable fractions is a health concern, meaning that species-level identification is warranted.

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%) wereBrevundimonas(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%), andMesorhizobium(1.1%), and the fungal taxa wereCryptococcus(12%) followed byAlternaria(9%),Aspergillus(7%),Candida(3%),Cladosporium(2.9%),Schizophyllum(1.6%),Fusarium(1.4%),Gleotinia(1.3%) andPenicillium(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.

Transgenic alfalfa (Medicago sativa L.) plants overexpressing the Arabidopsis ATP sulfurylase gene were generated using Agrobacterium-mediated genetic transformation to enhance their heavy metal accumulation efficiency. The ATP sulfurylase gene was cloned from Arabidopsis, following exposure to vanadium (V) and lead (Pb), and transferred into an Agrobacterium tumefaciens binary vector. This was co-cultivated with leaf explants of the alfalfa genotype Regen SY. Co-cultivated leaf explants were cultured on callus and somatic embryo induction medium, followed by regeneration medium for regenerating complete transgenic plants. The transgenic nature of the plants was confirmed using PCR and southern hybridization. The expression of Arabidopsis ATP sulfurylase gene in the transgenic plants was evaluated through RT-PCR. The selected transgenic lines showed increased tolerance to a mixture of five heavy metals and also demonstrated enhanced metal uptake ability under controlled conditions. The transgenic lines were fertile and did not exhibit any apparent morphological abnormality. The results of this study indicated an effective approach to improve the heavy metal accumulation ability of alfalfa plants which can then be used for the remediation of contaminated soil in arid regions.

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.

The role of rhizosphere microbial communities in the degradation of hydrocarbons remains poorly understood and is a field of active study. We used high throughput sequencing to explore the rhizosphere microbial diversity in the alfalfa and barley planted oil contaminated soil samples. The analysis of 16s rRNA sequences showed Proteobacteria to be the most enriched (45.9%) followed by Bacteriodetes (21.4%) and Actinobacteria (10.4%) phyla. The results also indicated differences in the microbial diversity among the oil contaminated planted soil samples. The oil contaminated planted soil samples showed a higher richness in the microbial flora when compared to that of untreated samples, as indicated by the Chao1 indices. However, the trend was different for the diversity measure, where oil contaminated barley planted soil samples showed slightly lower diversity indices. While the clustering of soil samples grouped the oil contaminated samples within and across the plant types, the clean sandy soil samples formed a separate group. The oil contaminated rhizosphere soil showed an enrichment of known oil-degrading genera, such as Alcanivorax and Aequorivita, later being specifically enriched in the contaminated soil samples planted with barley. Overall, we found a few well known oil-degrading bacterial groups to be enriched in the oil contaminated planted soil samples compared to the untreated samples. Further, phyla such as Thermi and Gemmatimonadetes showed an enrichment in the oil contaminated soil samples, indicating their potential role in hydrocarbon degradation. The findings of the current study will be useful in understanding the rhizosphere microflora responsible for oil degradation and thus can help in designing appropriate phytoremediation strategies for oil contaminated lands.

Rhanterium eppaposum Oliv. (Arfaj) the national plant of Kuwait, is a perennial bushy shrub and the national flower of Kuwait. It is one of the most palatable forage plants and an excellent performer for desert rehabilitation. Due to several anthropogenic activities and climate change, this native plant has suffered severe degradation. In order to formulate conservation strategies, the genetic diversity of Rhanterium, was assessed by inter simple sequence repeat (ISSR) markers. A total of 93 wild landraces distributed in six remote locations in Kuwait were tested using 17 primers (PIC = 0.31; polymorphism = 92.8 %) that produced 195 unambiguous and reproducible bands. Overall, diversity indices were high enough (Nei’s gene diversity (h') - 0.286; Shannon’s diversity index - 0.414) for these populations. AMOVA test returned a fixation coefficient (FST) of 0.116 (p = 0.0) with higher genetic variations detected within (88 %) the populations as compared to among (12 %) the populations. Satisfactory levels of genetic differentiation (GST = 0.186; G’ST = 0.250; D’ = 0.295) were recorded for R. eppaposum with a gene flow (NM) maintained at 1.9. The maximum genetic distances were observed between Maqwa and Mina Abdulla (Ds- 0.158; FST- 0.228) whereas minimum was between Al Kabd and SANR (Ds - 0.06; FST- 0.05). A weak Mantel’s correlation (r2 - 0.139; P = 0.014) between the six populations was also recorded. A narrow population structure was observed through the PCoA (2 groups), STRUCTURE (2 groups, K = 2) and SPLIT decomposition (1 cluster). The high genetic diversity within the populations are encouraging, however the low genetic differentiation raises concerns. The population structure should be improved by planting genetically diverse genotypes in areas of very low diversity to broaden its genetic base. Keywords: Conservation management, Genetic diversity, Molecular markers, Arfaj, Polymorphism

Developments in genetic engineering technology have led to an increase in number of food products that contain genetically engineered crops in the global market. However, due to lack of scientific studies, the presence of genetically modified organisms (GMOs) in the Kuwaiti food market is currently ambiguous. Foods both for human and animal consumption are being imported from countries that are known to produce GM food. Therefore, an attempt has been made to screen foods sold in the Kuwaiti market to detect GMOs in the food. For this purpose, samples collected from various markets in Kuwait have been screened by SYBR green-based real time polymerase chain reaction (RT-PCR) method. Further confirmation and GMO quantification was performed by TaqMan-based RT-PCR. Results indicated that a significant number of food commodities sold in Kuwait were tested positive for the presence of GMO. Interestingly, certain processed foods were tested positive for more than one transgenic events showing complex nature of GMOs in food samples. Results of this study clearly indicate the need for well-defined legislations and regulations on the marketing of approved GM food and its labeling to protect consumer's rights.

The main aim of the study was to test for the presence of Legionnaires’ disease-causing microorganisms in air-conditioned buildings in Kuwait using molecular technologies. For this purpose, 547 samples were collected from 38 cooling towers for the analysis of Legionella pneumophila. These samples included those from water (n = 178), air (n = 231), and swabs (n = 138). Out of the 547 samples, 226 (41 %) samples were presumptive positive for L. pneumophila, with L. pneumophila viable counts in the positive water samples ranging from 1 to 88 CFU/ml. Of the Legionella culture-positive samples, 204 isolates were examined by latex agglutination. These isolates were predominately identified as L. pneumophila serogroup (sg) 2–14. Using the Dresden panel of monoclonal antibodies, 74 representatives isolates were further serogrouped. Results showed that 51 % of the isolates belonged to serogroup 7 followed by 1 (18 %) and 3 (18 %). Serogroups 4 (4 %) and 10 (7 %) were isolated at a lower frequency, and two isolates could not be assigned to a serogroup. These results indicate the wide prevalence of L. pneumophila serogroup 7 as the predominant serogroup at the selected sampling sites. Furthermore, the 74 L. pneumophila (sg1 = 13; sg3 = 13; sg4 = 3; sg7 = 38; sg10 = 5; sgX = 2) isolates were genotyped using the seven gene protocol sequence-based typing (SBT) scheme developed by the European Working Group for Legionella Infections (EWGLI). The results show that Legionella isolates were discriminated into nine distinct sequence typing (ST) profiles, five of which were new to the SBT database of EWGLI. Additionally, all of the ST1 serogroup 1 isolates were of the OLDA/Oxford subgroup. These baseline data will form the basis for the development of a Legionella environmental surveillance program and used for future epidemiological investigations.