Silver nanoparticles biosynthesized from secondary metabolite producing marine actinobacteria and evaluation of their biomedical potential.

Biosynthesis of silver nanoparticles (AgNPs) from marine actinobacteria offers a promising avenue for exploring bacterial extracts as reducing and stabilizing agents. We report extracellular extracts of Rhodococcus rhodochrous (MOSEL-ME29) and Streptomyces sp. (MOSEL-ME28), identified by 16S rRNA gene sequencing for synthesis of AgNPs. Ultrafine silver nanoparticles were biosynthesized using the extracts of R. rhodochrous and Streptomyces sp. and their possible therapeutic applications were studied. The physicochemical properties of nanoparticles were established by HR-SEM/TEM, SAED, UV-Vis, EDS, XRD, and FTIR. UV-Vis spectra displayed characteristic absorption at 430 nm and 412 nm for AgNPs from Streptomyces sp. (S-AgNPs) and Rhodococcus sp. (R-AgNPs), respectively. HR-SEM/TEM, XRD, EDS analysis confirmed the spherical shape, crystalline nature, and elemental formation of silver. Crystallite or grain size was deduced as 5.52 nm for R-AgNPs and 35 nm for S-AgNPs. Zeta-potential indicated electrostatic negative charge for AgNPs, while FTIR revealed the presence of diverse functional groups. Disc diffusion assay indicated the broad-spectrum antibacterial potential of S-AgNPs with the maximum inhibition of B. subtilis while R-AgNPs revealed potency against P. aeruginosa at 10 µg/mL concentration. Biogenic AgNPs revealed antileishmanial activity and the IC50 was calculated as 164 µg/mL and 184 µg/mL for R-AgNPs and S-AgNPs respectively. Similarly, the R-AgNPs and S-AgNPs revealed anti-cancer potential against HepG2 and the IC50 was calculated as 49 µg/mL and 69 µg/mL for R-AgNPs and S-AgNPs, respectively. Moreover, the antioxidant activity showed significant results. MTT assay on RD cells, L20B cells, and Hep-2C indicated intensification in viability by reducing the concentration of R-AgNPs and S-AgNPs. The R-AgNPs and S-AgNPs inhibited sabin-like poliovirus (1TCID50 infection in RD cells). Furthermore, hemocompatibility at low concentrations has been confirmed. Hence, it is concluded that biogenic-AgNPs has the potential to be used in diverse biological applications and that the marine actinobacteria are an excellent resource for fabrication of AgNPs.

Improving Biosecurity in Pakistan: Current Efforts, Challenges, and Recommendations on a Multidimensional Management Strategy.

Rapidly advancing biotechnological research and easy access to biological information have created challenges for preventing their intentional misuse. Because of the dedicated efforts of science and policy leaders in Pakistan, the country is on the road to becoming a regional role model in ensuring biosafety and capacity for biosecurity. Although Pakistan has made remarkable efforts promoting a culture of biosafety and biosecurity, several constraints persist related to sustainable development and the expansion of the biosecurity capacity in Pakistan. Studies suggest there is little understanding of dual-use research of concern among the majority of life scientists in Pakistan. Further, Pakistan has yet to fully develop and implement approaches for risk assessment, frameworks for oversight and regulation, or tools and procedures for monitoring and reporting situations in which misuse of biological materials might occur. More data are needed to answer questions about key biosecurity hazards specific to the country or region and the procedures that may be required for periodic risk assessments. Integrating biosecurity into the existing infrastructure with limited resources is key challenge in implementation. More coordination is required among various stakeholders to develop an effective biosecurity framework. This article focuses on Pakistan's strides toward biosecurity preparedness in terms of protecting pathogens, toxins, or biological technologies with dual-use risks to prevent their malevolent use. We propose a multidimensional biosecurity management model at the national level to strengthen the biosecurity enterprise in Pakistan.

Genome sequence analysis and bioactivity profiling of marine-derived actinobacteria, Brevibacterium luteolum, and Cellulosimicrobium funkei.

Genome analysis gives important insights into the biosynthetic potential of marine actinobacteria. The genomes of two marine actinomycetes Brevibacterium luteolum MOSEL-ME10a and Cellulosimicrobium funkei MOSEL-ME6 were sequenced to identify the biosynthetic gene clusters (BGCs). Additionally, anti-proliferative, antioxidant, and enzyme inhibitory activities were studied in vitro. We report a total genome size of 2.77 Mb with GC content of 67.8% and 6.81 Mb with GC content of 69% for Brevibacterium sp. MOSEL-ME10a and Cellulosimicrobium sp. MOSEL-ME6, respectively. Biosynthetic gene clusters (BGCs) encoding different classes of natural products were predicted including terpenes, peptides, siderophores, ectoines, and bacteriocins. The bioactivity potential of crude extracts derived from these strains was evaluated. Notable anti-proliferative activity was observed against HepG2 cell line (hepatocellular carcinoma) with an IC50 value of 182 µg/mL for Brevibacterium sp. MOSEL-ME10a. Furthermore, antioxidant activity was assessed with IC50 values of 48.91 µg/mL and 102.5 µg/mL for Brevibacterium sp. MOSEL-ME10a and Cellulosimicrobium sp. MOSEL-ME6, respectively. Protein kinase inhibition potential was observed only for Brevibacterium sp. MOSEL-ME10a. Our study also reports lower amylase enzyme inhibition potential for both strains. Moreover, both crude extracts showed only slight-to-no toxic effect on erythrocytes at 400 µg/mL and below, indicating erythrocyte membrane stability. Our data present the genomic features revealing biosynthetic potential of marine actinobacteria as well as biological activities found in vitro.

Ethics, pandemic and environment; looking at the future of low middle income countries.

COVID-19 which started in Wuhan, China and swiftly expanded geographically worldwide, including to Low to Middle Income Countries (LMICs). This in turn raised numerous ethical concerns in preparedness, knowledge sharing, intellectual property rights, environmental health together with the serious constraints regarding readiness of health care systems in LMICs to respond to this enormous public health crisis. From the restrictions on public freedom and burgeoning socio-economic impacts to the rationing of scarce medical resources, the spread of COVID-19 is an extraordinary ethical dilemma for resource constrained nations with less developed health and research systems. In the current crisis, scientific knowledge and technology has an important role to play in effective response. Emergency preparedness is a shared responsibility of all countries with a moral obligation to support each other. This review discusses the ethical concerns regarding the national capacities and response strategies in LMICs to deal with the COVID-19 pandemic as well as the deep link between the environment and the increasing risk of pandemics.

Genome of Serratia nematodiphila MB307 offers unique insights into its diverse traits.

A pigment-producing species of Serratia was isolated from the rhizosphere of a heavy metal resistant Cannabis sativa plant growing in effluent-affected soil of Hattar Industrial Estate, Haripur, Pakistan. Here, we report the genome sequence of this bacterium, which has been identified as Serratia nematodiphila on the basis of whole genome comparison using the OrthoANI classification scheme. The bacterium exhibited diverse traits, including plant growth promotion, antimicrobial, bioremediation, and pollutant tolerance capabilities including metal tolerance, azo dye degradation, ibuprofen degradation, etc. Plant growth-promoting exoenzyme production as well as phosphate solubilisation properties were observed. Genes for phosphate solubilisation, siderophore production, and chitin destruction were identified in addition to other industrially important enzymes like nitrilase and lipase. Secondary metabolite producing apparatus for high value chemicals in the whole genome was also analysed. The number of antibiotic resistance genes was then profiled in silico, through a match with Antibiotic Resistant Gene and CAR database. This is the first report of a S. nematodiphila genome from a polluted environment. This could significantly contribute to the understanding of pollution tolerance, antibiotic resistance, association with nematodes, production of bio-pesticide, and their role in plant growth promotion.

Ethical Issues Regarding CRISPR Mediated Genome Editing.

CRISPR-Cas9 has emerged as a simple, precise and most rapid genome editing technology. With a number of promising applications ranging from agriculture and environment to clinical therapeutics, it is greatly transforming the field of molecular biology. However, there are certain ethical, moral and safety concerns related to the attractive applications of this technique. The most contentious issues concerning human germline modifications are the challenges to human safety and morality such as risk of unforeseen, undesirable effects in clinical applications particularly to correct or prevent genetic diseases, matter of informed consent and the risk of exploitation for eugenics. Stringent regulations and guidelines as well as worldwide debate and awareness are required to ensure responsible and wise use of CRISPR mediated genome editing technology. There is a need for an extensive dialogue among scientists, ethicists, industrialists and policy makers on its societal implications. The opinion of different elements of the society including the general public as well as religious scholars is also critical. In countries with existing legislative framework, it might be appropriate to allow CRISPR based research to proceed with proper justification. However, much anticipated future clinical applications must be strictly regulated with newly established regulations.

An assessment on the role of endophytic microbes in the therapeutic potential of Fagonia indica.

BACKGROUND: Natural products of animals, plants and microbes are potential source of important chemical compounds, with diverse applications including therapeutics. Endophytic bacteria that are especially associated with medicinal plants presents a reservoir of therapeutic compounds. Fagonia indica has been recently investigated by numerous researchers because of its striking therapeutic potential especially in cancer. It is also reported that endophytes play a vital role in the biosynthesis of various metabolites; therefore we believe that endophytes associated with F. indica are of crucial importance in this regard. The present study aims successful isolation, molecular identification of endophytic bacteria and their screening for bioactive metabolites quantification and in vitro pharmacological activities. METHODS: 16S rRNA gene sequencing was used for the identification of isolated endophytic bacteria. Methanolic extracts were evaluated for total phenolic contents (TPC), total flavonoids contents (TFC), DPPH free radical scavenging activity, reducing power and total anti-oxidant assays were performed. And also screened for antibacterial and antifungal activities by disc diffusion method and their MIC were calculated by broth dilution method using microplate reader. Further, standard protocols were followed for antileishmanial activity and protein kinase inhibition. Analysis and statistics were performed using SPSS, Table curve and Origin 8.5 for graphs. RESULTS: Bacterial strains belonging to various genera (Bacillus, Enterobacter, Pantoea, Erwinia and Stenotrophomonas) were isolated and identified. Total phenolic contents and total flavonoids contents varies among all the bacterial extracts respectively in which Bacillus subtilis showed high phenolic contents 243 µg/mg of gallic acid equivalents (GAE) and Stenotrophomonas maltophilia showed high flavonoids contents 15.9 µg/mg quercitin equivalents (QA), total antioxidant capacity (TAC) 37.6 µg/mg of extract, reducing power (RP) 206 µg/mg of extract and 2, 2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging activity with 98.7 µg/mL IC50 value. Although all the extracts tested were active to inhibit growth of selected pathogenic microbes (bacteria and fungi), but significant antibacterial activity was observed against Klebsiella pneumonia and B. subtilis. An Enterobacter cloaca was active against Leishmania tropica with IC50 value of 1.4 µg/mg extracts. B. subtilis and Bacillus tequilensis correspondingly exhibit significant protein kinase inhibition of 47 ± 0.72 and 42 ± 1.21 mm bald zones, indicating anti-infective and antitumor potential. CONCLUSIONS: Our findings revealed that crude extracts of selected endophytic bacteria from F. indica possess excellent biological activities indicating their potential as an important source of antibiotics (antifungal, antibacterial) compounds.

Emerging Viral Infections in Pakistan: Issues, Concerns, and Future Prospects.

Emerging infectious diseases pose a serious threat to public health security; this is especially true in the underdeveloped world because of limited resources to combat them. These emerging pathogens are characterized by a novel mode of pathogenesis and, in some cases, a broad host range. Over the past few decades, Pakistan has suffered a great deal from infectious diseases such as dengue, Crimean-Congo fever, hepatitis, measles, and polio. Changing climate conditions, environmental degradation, global warming, loss of biodiversity, and other ecological determinants have a direct effect on these diseases and result in the emergence and reemergence of infectious entities. The causes of such disease outbreaks are complex and often not well understood. Dealing with an outbreak requires an integrated and coordinated approach, with decision making by various state departments. Stringent biosecurity and biosafety protocols can help to reduce the chances of infection dissemination. In order to mitigate the risks associated with emerging pathogens, there is a greater need to understand the interactions of pathogen-host-environment, to monitor molecular evolution and genomic surveillance, and to facilitate the gearing up of scientists across the globe to control these emerging diseases. This article reviews recent outbreaks in Pakistan and challenges for the development of an agile healthcare setup in the country.