Studies of plant colonisation by closely related Bacillus amyloliquefaciens biocontrol agents using strain specific quantitative PCR assays.

Certain strains of Bacillus amyloliquefaciens can colonize plants and improve growth and stress management. In order to study these effects, bacterial growth dynamics on plants and in the rhizosphere are of interest calling for specific analytical tools. For that purpose, quantitative real-time PCR (qPCR) assays were developed in order to differentiate among three closely related B. amyloliquefaciens subsp. plantarum strains (UCMB5033, UCMB5036, UCMB5113) and to determine their levels with high accuracy. Oligonucleotide primers were designed for strain unique gene sequences and used for SYBR green based qPCR analysis. Standard curves covered a wide linear range (10(6)) of DNA amounts with the lowest detection level at 50 fg. Post-reaction melting curve analysis showed only a single product. Accurate threshold cycles were obtained, even in the presence of high excess of related Bacillus strains and total bacterial DNA from soil. Analysis of Bacillus colonisation after seed treatment of two oilseed rape cultivars (Oase and Ritz) grown on agar support showed a time dependent effect but that the bacteria mostly were found on root tissues and little on green tissues. The colonisation on plants grown in soil varied among the Bacillus strains where Oase seemed to house more bacteria than Ritz. Applied as a mixture, all three Bacillus strains co-existed on the roots of plants grown in soil. The qPCR assay in combination with other techniques will be a powerful tool to study plant interactions of these B. amyloliquefaciens biocontrol agents to further understand the requirements for successful interactions and improvement of plant properties.

Linkage of natural resources, economic policies, urbanization, and the environmental Kuznets curve.

Natural resource rents (NRR) and economic policies are considerably studied to determine ecological footprints. Currently, due to global uncertainty, renewable energy adoption, and increasing urbanization, every economy is facing challenges to control its ecological footprints. The available literature on the said linkages in the emerging seven economies is inconclusive. Therefore, this study is designed to re-estimate the linkages of NRR, urbanization (URB), economic policy uncertainty (EPU), energy structure (ES), and EFP under the "Environment Kuznets Curve (EKC) hypothesis." Data from 1992 to 2020 is used for empirical evidence, along with robust econometric calculations. The EKC hypothesis does not apply to the E7 economies, according to the findings. The energy structure is assisting in limiting ecological footprints and hence aids in environmental cleanup. The role of NRR, EPU, and URB in limiting the EF, on the other hand, is not encouraging. To minimize environmental degradation, emerging economies should reconsider their economic policies, natural resource rents, and rapid urbanization.

Impact of Fear of COVID-19 Pandemic on Job Insecurity and Subjective Well-Being.

Earlier research on the novel coronavirus (COVID-19) mainly focused on fear and its psychological impact on frontline health workers. However, the uncertainty of COVID-19, job insecurity and its effects on non-frontline employees' perception of their well-being is rarely explored. This study aims to assess the relationship between subjective well-being and the fear of the COVID-19 outbreak. Furthermore, this study explores how employees' awareness of their job insecurity influences this relationship. Structural Equation Modelling (SEM) and Confirmatory Factor Analysis (CFA), and Spearman's correlation analyses were used to examine the significance of the relationships between fear of COVID-19 and subjective well-being and the moderating role of job insecurity in this relationship. The findings indicate that greater fear of COVID-19 is associated with a lower level of perceived subjective well-being. However, a higher perception of fear of COVID-19 and its interaction with job insecurity are associated with a higher risk of employees' well-being. Our study highlights the factors such as fear of COVID-19 and job insecurities that undermine their well-being. Our study provdies practical implications for employees', managers, and healthcare policymakers to adopt effective strategies to reduce stress among employees. These recommendations include using practical tactics to safeguard the employees health and jobs while effectively coping with the pandemic.

Leadership Styles and Organizational Citizenship Behavior for the Environment: The Mediating Role of Self-Efficacy and Psychological Ownership.

Change and environmental patterns are having an immense effect upon the world. Businesses, communities, and even individuals are struggling to perform their role within environmental protection. This paper investigates the role of leadership styles on organizational citizenship behavior for the environment (OCBE) directly and through the mediation of self-efficacy and psychological ownership. The survey technique was used to collect the data from Chinese banking, insurance, medicine, and teaching service sector employees for the current study. The reliability and validity of the scale items were tested. This study used AMOS-SEM for data analysis and testing the developed hypotheses. The empirical results confirmed that responsible, inclusive, authentic, and supportive leadership styles positively impact employees' OCBE. The results further confirm that self-efficacy and psychological ownership act as mediators between leadership and OCBE. The current study widens our understanding of leadership styles and their impact on OCBE, along with limitations associated with the study and future guidelines for investigators.

Role of Responsible Leadership for Organizational Citizenship Behavior for the Environment in Light of Psychological Ownership and Employee Environmental Commitment: A Moderated Mediation Model.

The world is looking toward organizations for social responsibility to contribute to a sustainable environment. Employees' organizational citizenship behavior for the environment (OCBE) is a voluntary environmental-oriented behavior that is important for organizations' environmental performance. Based on social learning theory, this study examined the effects of responsible leadership in connection with OCBE by using a sample of 520 employees in the manufacturing and service sectors in China including engine manufacturing, petroleum plants, banking, and insurance sector organizations. Further, the roles of psychological ownership and employee environmental commitment were used as mediators and moderators simultaneously. The direct, mediation, and moderation model results exposed a positive relationship between responsible leadership and OCBE via employee psychological ownership and employee environmental commitment. The study also revealed that the indirect effect is stronger when employees hold a higher employee environmental commitment. The theoretical and practical implications for environmental sustainability in respect of organizations as well as future research directions are discussed.

Genome-Guided Analysis of Clostridium ultunense and Comparative Genomics Reveal Different Strategies for Acetate Oxidation and Energy Conservation in Syntrophic Acetate-Oxidising Bacteria.

Syntrophic acetate oxidation operates close to the thermodynamic equilibrium and very little is known about the participating organisms and their metabolism. Clostridium ultunense is one of the most abundant syntrophic acetate-oxidising bacteria (SAOB) that are found in engineered biogas processes operating with high ammonia concentrations. It has been proven to oxidise acetate in cooperation with hydrogenotrophic methanogens. There is evidence that the Wood-Ljungdahl (WL) pathway plays an important role in acetate oxidation. In this study, we analysed the physiological and metabolic capacities of C. ultunense strain Esp and strain BST on genome scale and conducted a comparative study of all the known characterised SAOB, namely Syntrophaceticus schinkii, Thermacetogenium phaeum, Tepidanaerobacter acetatoxydans, and Pseudothermotoga lettingae. The results clearly indicated physiological robustness to be beneficial for anaerobic digestion environments and revealed unexpected metabolic diversity with respect to acetate oxidation and energy conservation systems. Unlike S. schinkii and Th. phaeum, C. ultunense clearly does not employ the oxidative WL pathway for acetate oxidation, as its genome (and that of P. lettingae) lack important key genes. In both of those species, a proton motive force is likely formed by chemical protons involving putative electron-bifurcating [Fe-Fe] hydrogenases rather than proton pumps. No genes encoding a respiratory Ech (energy-converting hydrogenase), as involved in energy conservation in Th. phaeum and S. schinkii, were identified in C. ultunense and P. lettingae. Moreover, two respiratory complexes sharing similarities to the proton-translocating ferredoxin:NAD⁺ oxidoreductase (Rnf) and the Na⁺ pumping NADH:quinone hydrogenase (NQR) were predicted. These might form a respiratory chain that is involved in the reduction of electron acceptors rather than protons. However, involvement of these complexes in acetate oxidation in C. ultunense and P. lettingae needs further study. This genome-based comparison provides a solid platform for future meta-proteomics and meta-transcriptomics studies and for metabolic engineering, control, and monitoring of SAOB.

Measurement of Radioactivity in the Granites of Pakistan: A Review.

All human beings are exposed to primordial radiation. The main source of primordial radiation has been naturally radioactive building materials. In Pakistan, among building materials, granites are most commonly used. This paper reviews the previous studies conducted for the estimation of natural radioactivity of granites from Rustam-Koga, Shewa-Shahbazgarhi, Bunair, Mansehra, and Nagarparkar. The techniques used for these studies include gamma spectrometry and field surveys. To assess the radiation hazards associated with granites of Pakistan, primordial radionuclides and corresponding radiation hazards have been estimated. Most of these granites are found to be safe except Rustam-Koga granite, which presents high excess lifetime cancer risk.

Complete genome sequence of Methanoculleus bourgensis strain MAB1, the syntrophic partner of mesophilic acetate-oxidising bacteria (SAOB).

Methanoculleus bourgensis strain MAB1 has been identified as the hydrogenotrophic partner of mesophilic acetate-oxidising bacteria, a syntrophic relationship operating close to the thermodynamic equilibrium and of considerable importance in ammonia-rich engineered biogas processes. Methanoculleus bourgensis strain MAB1 belongs to the order Methanomicrobiales, family Methanomicrobiaceae, within the phylum Euryarchaeota. The genome shows a total size of 2,859,299 bp encoding 3450 predicted protein-encoding genes, of which only 1472 (43 %) have been assigned tentative functions. The genome encodes further 44 tRNA genes and three rRNA genes (5S, 16S and 23S rRNA). This study presents assembling and annotation features as well as genomic traits related to ammonia tolerance and methanogenesis.

Genome-Guided Analysis and Whole Transcriptome Profiling of the Mesophilic Syntrophic Acetate Oxidising Bacterium Syntrophaceticus schinkii.

Syntrophaceticus schinkii is a mesophilic, anaerobic bacterium capable of oxidising acetate to CO2 and H2 in intimate association with a methanogenic partner, a syntrophic relationship which operates close to the energetic limits of microbial life. Syntrophaceticus schinkii has been identified as a key organism in engineered methane-producing processes relying on syntrophic acetate oxidation as the main methane-producing pathway. However, due to strict cultivation requirements and difficulties in reconstituting the thermodynamically unfavourable acetate oxidation, the physiology of this functional group is poorly understood. Genome-guided and whole transcriptome analyses performed in the present study provide new insights into habitat adaptation, syntrophic acetate oxidation and energy conservation. The working draft genome of Syntrophaceticus schinkii indicates limited metabolic capacities, with lack of organic nutrient uptake systems, chemotactic machineries, carbon catabolite repression and incomplete biosynthesis pathways. Ech hydrogenase, [FeFe] hydrogenases, [NiFe] hydrogenases, F1F0-ATP synthase and membrane-bound and cytoplasmic formate dehydrogenases were found clearly expressed, whereas Rnf and a predicted oxidoreductase/heterodisulphide reductase complex, both found encoded in the genome, were not expressed under syntrophic growth condition. A transporter sharing similarities to the high-affinity acetate transporters of aceticlastic methanogens was also found expressed, suggesting that Syntrophaceticus schinkii can potentially compete with methanogens for acetate. Acetate oxidation seems to proceed via the Wood-Ljungdahl pathway as all genes involved in this pathway were highly expressed. This study shows that Syntrophaceticus schinkii is a highly specialised, habitat-adapted organism relying on syntrophic acetate oxidation rather than metabolic versatility. By expanding its complement of respiratory complexes, it might overcome limiting bioenergetic barriers, and drive efficient energy conservation from reactions operating close to the thermodynamic equilibrium, which might enable S. schinkii to occupy the same niche as the aceticlastic methanogens. The knowledge gained here will help specify process conditions supporting efficient and robust biogas production and will help identify mechanisms important for the syntrophic lifestyle.

Antihypertensive nano-ceuticales based on chitosan biopolymer: Physico-chemical evaluation and release kinetics.

Prime risk factor behind cardiovascular associated mortality and morbidity is hypertension. The main challenge with antihypertensive (AHT) drug therapy is their extreme hydrophobic nature and very low oral bio-availability; which result into higher dosage/frequency and associated side effects of drugs. The main objective of this study was to fabricate AHT nano-ceuticals in hydrophilic carriers of natural origin to improve drugs' solubility, protection and sustained release. AHT nano-carrier systems (NCS) encapsulating captopril, amlodipine and valsartan were fabricated using chitosan (CS) polymer by ionic gelation assisted ultra-sonication method. Drug encapsulation efficiencies of 92±1.6%, 91±0.9% and 87±0.5% were observed for captopril, valsartan and amlodipine respectively. Scanning electron microscopy (SEM) based analysis had revealed that captopril loaded polymeric NCS were regular, smooth and without any agglomeration. FTIR analyses of drug loaded and empty NCS demonstrated that drugs were molecularly dispersed inside the nanoparticles via week hydrogen bonding. Captopril and valsartan have demonstrated grafting reaction with N-H group of chitosan. Zeta sizer results had confirmed that average size of chitosan nanoparticles was below 100 nm. Encapsulation of captopril had reduced the surface charge value from +52.6±4.8 to +46.5±5.2 mV. Controlled release evaluation of highly encapsulated drug captopril had revealed a slow release in vitro from NCS in physiological buffer. Thus, here reported innovative AHT nano-ceuticals of polymeric origin can improve the oral administration of currently available hydrophobic drugs while providing the extended-release function.

Perception of gastroesophageal reflux disease in urban population in Pakistan.

OBJECTIVE: To determine the frequency and health care seeking behavior of patients with gastroesophageal reflux symptoms using American College of Gastroenterology (ACG) questionnaire for gastroesophageal reflux disease (GERD). DESIGN: Cross-sectional study. PLACE AND DURATION OF STUDY: Section of Gastroenterology, Department of Medicine, The Aga Khan University Hospital, Karachi, from April 2002 to November 2003. PATIENTS AND METHODS: A total of 1267 persons above 15 years of age were interviewed using a modified questionnaire for GERD from the American College of Gastroenterology (ACG), which previously validated in our section as a screening tool in an urban population in Pakistan. RESULTS: Nine hundred and sixty-three individuals responded to the questionnaire. Mean age of the surveyed sample was 24+/-9.3 years. Using the ACG criteria, the overall frequency of gastroesophageal reflux symptoms was 24% (228/963) with 58% (133/228) males and 42% (95/228) females. Symptoms experienced frequently were reflux from the stomach, 58% (133/228), with a bitter, acid taste in the mouth. Seventy-four percent (169/228) experienced this postprandially. Symptoms were associated with spicy fried food in 71% (161/228). Sixty-seven percent (152/228) did not seek health care advice, 32% (74/228) practiced self-medications and 33% (76/228) consulted a physician. CONCLUSION: Gastroesophageal reflux is a common problem in the urban population of Pakistan.

Working draft genome sequence of the mesophilic acetate oxidizing bacterium Syntrophaceticus schinkii strain Sp3.

Syntrophaceticus schinkii strain Sp3 is a mesophilic syntrophic acetate oxidizing bacterium, belonging to the Clostridia class within the phylum Firmicutes, originally isolated from a mesophilic methanogenic digester. It has been shown to oxidize acetate in co-cultivation with hydrogenotrophic methanogens forming methane. The draft genome shows a total size of 3,196,921 bp, encoding 3,688 open reading frames, which includes 3,445 predicted protein-encoding genes and 55 RNA genes. Here, we are presenting assembly and annotation features as well as basic genomic properties of the type strain Sp3.

Draft genome sequence of 'Treponema phagedenis' strain V1, isolated from bovine digital dermatitis.

'Treponema phagedenis' is considered to be a key agent in the pathogenesis of bovine digital dermatitis, an infectious foot condition of economic and animal welfare importance. We hereby report the draft sequence of 'T. phagedenis' strain V1. The draft genome assembly consists of 51 scaffolds comprising 3,129,551 bp and a GC-content of 39.9 %. Putative pathogenicity related factors have been identified in the genome that can be used in future studies to gain insight into the pathogenic mechanisms of 'T. phagedenis'.

Genome-guided analysis of physiological capacities of Tepidanaerobacter acetatoxydans provides insights into environmental adaptations and syntrophic acetate oxidation.

This paper describes the genome-based analysis of Tepidanaerobacter acetatoxydans strain Re1, a syntrophic acetate-oxidising bacterium (SAOB). Principal issues such as environmental adaptations, metabolic capacities, and energy conserving systems have been investigated and the potential consequences for syntrophic acetate oxidation discussed. Briefly, in pure culture, T. acetatoxydans grows with different organic compounds and produces acetate as the main product. In a syntrophic consortium with a hydrogenotrophic methanogen, it can also reverse its metabolism and instead convert acetate to formate/H2 and CO2. It can only proceed if the product formed is continuously removed. This process generates a very small amount of energy that is scarcely enough for growth, which makes this particular syntrophy of special interest. As a crucial member of the biogas-producing community in ammonium-rich engineered AD processes, genomic features conferring ammonium resistance, bacterial defense, oxygen and temperature tolerance were found, as well as attributes related to biofilm formation and flocculation. It is likely that T. acetatoxydans can form an electrochemical gradient by putative electron-bifurcating Rnf complex and [Fe-Fe] hydrogenases, as observed in other acetogens. However, genomic deficiencies related to acetogenic metabolism and anaerobic respiration were discovered, such as the lack of formate dehydrogenase and F1F0 ATP synthase. This has potential consequences for the metabolic pathways used under SAO and non-SAO conditions. The two complete sets of bacteriophage genomes, which were found to be encoded in the genome, are also worthy of mention.

Complete genome sequence of a plant associated bacterium Bacillus amyloliquefaciens subsp. plantarum UCMB5033.

Bacillus amyloliquefaciens subsp. plantarum UCMB5033 is of special interest for its ability to promote host plant growth through production of stimulating compounds and suppression of soil borne pathogens by synthesizing antibacterial and antifungal metabolites or priming plant defense as induced systemic resistance. The genome of B. amyloliquefaciens UCMB5033 comprises a 4,071,167 bp long circular chromosome that consists of 3,912 protein-coding genes, 86 tRNA genes and 10 rRNA operons.

Genome analysis of Bacillus amyloliquefaciens Subsp. plantarum UCMB5113: a rhizobacterium that improves plant growth and stress management.

The Bacillus amyloliquefaciens subsp. plantarum strain UCMB5113 is a Gram-positive rhizobacterium that can colonize plant roots and stimulate plant growth and defense based on unknown mechanisms. This reinforcement of plants may provide protection to various forms of biotic and abiotic stress. To determine the genetic traits involved in the mechanism of plant-bacteria association, the genome sequence of UCMB5113 was obtained by assembling paired-end Illumina reads. The assembled chromosome of 3,889,532 bp was predicted to encode 3,656 proteins. Genes that potentially contribute to plant growth promotion such as indole-3-acetic acid (IAA) biosynthesis, acetoin synthesis and siderophore production were identified. Moreover, annotation identified putative genes responsible for non-ribosomal synthesis of secondary metabolites and genes supporting environment fitness of UCMB5113 including drug and metal resistance. A large number of genes encoding a diverse set of secretory proteins, enzymes of primary and secondary metabolism and carbohydrate active enzymes were found which reflect a high capacity to degrade various rhizosphere macromolecules. Additionally, many predicted membrane transporters provides the bacterium with efficient uptake capabilities of several nutrients. Although, UCMB5113 has the possibility to produce antibiotics and biosurfactants, the protective effect of plants to pathogens seems to be indirect and due to priming of plant induced systemic resistance. The availability of the genome enables identification of genes and their function underpinning beneficial interactions of UCMB5113 with plants.

Study of natural radioactivity in Mansehra granite, Pakistan: environmental concerns.

A part of Mansehra Granite was selected for the assessment of radiological hazards. The average activity concentrations of (226)Ra, (232)Th and (40)K were found to be 27.32, 50.07 and 953.10 Bq kg(-1), respectively. These values are in the median range when compared with the granites around the world. Radiological hazard indices and annual effective doses were estimated. All of these indices were found to be within the criterion limits except outdoor external dose (82.38 nGy h(-1)) and indoor external dose (156.04 nGy h(-1)), which are higher than the world's average background levels of 51 and 55 nGy h(-1), respectively. These values correspond to an average annual effective dose of 0.867 mSv y(-1), which is less than the criterion limit of 1 mSv y(-1) (ICRP-103). Some localities in the Mansehra city have annual effective dose higher than the limit of 1 mSv y(-1). Overall, the Mansehra Granite does not pose any significant radiological health hazard in the outdoor or indoor.

First Genome Sequence of a Syntrophic Acetate-Oxidizing Bacterium, Tepidanaerobacter acetatoxydans Strain Re1.

Syntrophic acetate-oxidizing bacteria (SAOB) have been identified as key organisms for efficient biogas production from protein-rich materials. Tepidanaerobacter acetatoxydans is the first reported SAOB for which the genome has been sequenced. Genome analysis will aid us in understanding the mechanisms regulating syntrophy, particularly energy-conserving and electron transfer mechanisms.

Draft Genome Sequence of Clostridium ultunense Strain Esp, a Syntrophic Acetate-Oxidizing Bacterium.

Clostridium ultunense strain Esp belongs to the functional group of syntrophic acetate-oxidizing bacteria (SAOB), which have been identified as key organisms for efficient biogas production from protein-rich materials. Genome analysis and comparative genomics might aid us to define physiological features that are essential for maintaining this particular syntrophic lifestyle.

Genome Sequence of a Plant-Associated Bacterium, Bacillus amyloliquefaciens Strain UCMB5036.

We announce here the genome sequence of Bacillus amyloliquefaciens strain UCMB5036, a plant growth-promoting bacterium isolated from a cotton plant. Its genome contains gene clusters involved in nonribosomal synthesis of secondary metabolites known for their antimicrobial activities. The availability of this genome will provide novel insights into plant-bacterium-associated activities.