ISOLATION, CHARACTERIZATION AND ANTIBACTERIAL SREENING OF ANTIBIOTICS PRODUCED FROM STREPTOMYCES ISOLATED FROM DUMPSITE SOILS IN ILORIN, NORTH CENTRAL NIGERIA

Antibiotics are one of the most exploited metabolites produced by soil actinomycetes. This study isolated fifteen actinomycetes (A1 – A15) from dumpsite soils within Ilorin metropolis & screened them for antibacterial activity. Isolates were identified with morphology & biochemical characteristics.Their activity against clinical bacteria (Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae & Staphylococcus aureus) was determined. Their bioactive metabolites after fermentation was assayed for by agar well diffusion & compared with reference antibiotics. Molecular analysis of two most active isolates was done. The most active metabolite was subjected to GCMS. All isolates were identified as Streptomyces, 7 isolates exhibited activity against 3 test bacteria with highest (10.5mm±0.35) by Streptomyces griseoplanus while S. aureus demonstrated total resistance to all isolates. Secondary activity revealed 7 of the metabolites as effective against the bacteria with highest activity (28.5±1.04mm) by S. sparsogenes showing similar activity with Cefuroxime (28.01±0.01mm). The two most active isolates were Streptomyces griseoplanus strain NR725RL-ISP 5009 & Streptomyces sparsogenes strain NBRC 1308616S & RNA nucleotide similarity revealed close phylogenic relationship. GC-MS revealed10 compounds in S. sparsogenes strain NBRC 1308616S metabolite. Streptomyces from dumpsite soils within Ilorin produced bioactive compounds against clinical test bacteria and thus could be potential sources of antibiotics.

The Distribution of COVID 19 based on Phylogeny Construction in Silico Sequences SARS-CoV-2 RNA at Genbank NCBI

The Covid-19 pandemic, due to severe acute respiratory coronavirus (SARS-CoV-2) virus, has an effect on human civilization today. With high fatality infections, SARS Covid-19 has influenced the global economic, socio-cultural, and even political order. This study aims to construct the phylogeny of the SARS corona virus that causes Covid-19 in various countries in the world by using the SARS Covid-19 gene database from the NCBI GenBank. The results of this study can trace the origin of SARS Covid-19, which is then called SARS-CoV-2, the gene characteristics, and the evolutionary relationship of these genes to various countries in the world. This research uses in silico method with gene sequence sources from the NCBI GenBank (www.ncbi.nih.gov). A total of 433 SARS Covid-19 sequences reported by 21 countries as of April 2rd, 2020 were the subject of the study. Sequences representing each country were analyzed using the MEGA 7.0 program. The results showed that the phylogeny trees formed were obtained by 2 main monophyletic groups. The first major monophyletic group consisted of 11 nodes, with 19 SARS-CoV-2 gene sequences from 23 countries. The second major monophyletic group consisted of 5 nodes with 5 countries of origin of SARS-CoV sequence 19. The spread of Covid-19 from the epicenter in Wuhan, China to the world has taken place randomly. This has happened because of the migration of people from the Chinese epicenter. The location of the countries adjacent to China did not determine the closest phylogenic relationship. The number of phylogenetic nodes formed showed mutases which caused very high variations of the SARS CoV 2 RNA gene sequence. The results of this study reinforce that efforts to limit the spread of human viruses to humans must be done. The presence of sequences from China in the 2 main monophyletic groups confirms that this virus originated in the Chinese epicenter.

A Proposal for the Classification and Nomenclature of Hand, Foot and Mouth Disease-Related Enteroviruses

Background: Enterovirus has diverged into many types, some of which cause hand, foot and mouth disease (HFMD) in children. The predominant enterovirus types associated with HFMD are EVA71, CVA16, CVA6 and CVA10. Subtyping of these enteroviruses is crucial to HFMD surveillance. Because of lacking proper and uniform criteria and being based on partial VP1 sequences, however, current classification resulted in some confusing and conflicting results. Method: We reclassified EVA71, CVA16, CVA6 and CVA10 using a combined criteria of phylogenic relationship and genetic distance. Results: Using the combined criteria, we classified EVA71 into seven genotypes of A–G, CVA16 and CVA6 into three subtypes of A-C, and CVA10 into nine subtypes/sub-subtypes of A-G, H1 and H2, and identified eight unclassified subtypes that lack genomic sequences. The mean genetic divergence was 15.5-33.8% between subtypes, 12-15% between sub-subtypes, and less than 12% within subtypes/sub-subtypes. In addition, we identified two new EVA71 inter-subtype recombinants RF01_CG and RF02_CG and demonstrated that EVA71 subtypes D and F and CVA10 subtype B experienced inter-subtype recombination events during early evolution. Conclusions: The new nomenclature proposal provides a reasonable framework for proper classification of enteroviruses, which will be useful for epidemiological surveillance of HFMD, disease management, and vaccine development.

Human URAT1/SLC22A12 gene promoter is regulated by 27-hydroxycholesterol through estrogen response elements

Elevated levels of uric acid, a metabolite of purine in humans, is related to various diseases, such as gout, atherosclerosis and renal dysfunction. The excretion and reabsorption of uric acid to/from urine is tightly regulated by uric acid transporters. The amino acid sequences of uric acid reabsorption transporters, URAT1/SLC22A12, OAT4/SLC22A11, and OAT10/SLC22A13, share closer phylogenic relationship, whereas the gene promoter sequences are distant phylogenic relationship. Through the single-cell RNA-sequencing analysis of an adult human kidney, we found that only a small number of cells express these transporters, despite their role in the regulation of serum uric acid levels. Transcriptional motif analysis on these transporter genes, revealed that the URAT1/SLC22A12 gene promoter displayed the most conserved estrogen response elements (EREs) among the three transporters. The endogenous selective estrogen receptor modulator (SERM) 27-hydroxycholesterol (27HC) had positive effects on the transcriptional activity of URAT1/SLC22A12. We also found that 27HC increased the protein and gene expression of URAT1/SLC22A12 in mouse kidneys and human kidney organoids, respectively. These results strongly suggest the role of 27HC for URAT1/SLC22A12 expression in renal proximal tubules and upregulation of serum uric acid levels and also show the relationship between cholesterol metabolism and serum uric acid regulation.Significance StatementThe elevated levels of serum uric acid cause various diseases, and the excretion/reabsorption of uric acid to/from urine is tightly regulated by the uric acid transporters. We found that despite the role in serum uric acid regulation, only a small number of cells express URAT1/SLC22A12. We also found that URAT1/SLC22A12 gene promoter region has effective estrogen response elements, and endogenous selective estrogen receptor (ER) modulator 27-hydroxycholesterol (27HC) increased URAT1/SLC22A12 expression in the mice kidneys and human kidney organoids. These suggest that 27HC increases URAT1/SLC22A12 expression and upregulate serum uric acid levels. Since 27HC connects cholesterol metabolism, our study indicates the important link between cholesterol metabolism and serum uric acid regulation, and also provides a novel therapeutic approach to hyperuricemia.

ORTHOSCOPE Analysis Reveals the Presence of the Cellulose Synthase Gene in All Tunicate Genomes but Not in Other Animal Genomes

Tunicates or urochordates—comprising ascidians, larvaceans, and salps—are the only metazoans that can synthesize cellulose, a biological function usually associated with bacteria and plants but not animals. Tunicate cellulose or tunicine is a major component of the outer acellular coverage (tunic) of the entire body of these organisms. Previous studies have suggested that the prokaryotic cellulose synthase gene (CesA) was horizontally transferred into the genome of a tunicate ancestor. However, no convenient tools have been devised to determine whether only tunicates harbor CesA. ORTHOSCOPE is a recently developed tool used to identify orthologous genes and to examine the phylogenic relationship of molecules within major metazoan taxa. The present analysis with this tool revealed the presence of CesA orthologs in all sequenced tunicate genomes but an absence in other metazoan genomes. This supports an evolutionary origin of animal cellulose and provides insights into the evolution of this animal taxon.