16S-Metagenomic data profiling of the bacterial community in ready-to-eat vegetable salads sold in Kampala, Uganda.

This article presents a dataset on bacterial community structure associated with Ready-to-eat (RTE) vegetable salads sold in Kampala City, Uganda. The Illumina Miseq sequencing of 16S rRNA gene amplicon unveiled the bacterial communities and generated a metagenomic library from RTE vegetable salads to understand the diversities and distribution. The metagenome contained a total of 23,805 sequences with 35,420 Taxonomic units (OTUs). Metagenome sequence information is obtainable at NCBI under the Bioproject assigned accession number PRJNA1064313. Taxonomic hits distribution from VSEARCH analysis at phylum level classification of NN-3 discovered predominantly Proteobacteria (65.34%) followed by Firmicutes (31.60%) and Bacteroidota (0.14%). Deinococcota (0.01%) and Planctomycetota (0.01%) were also detected. Also, VSEARCH-assisted analysis of NN-4 detected a higher prevalence of Firmicutes (65.68%) than Proteobacteria (33.25%), while Bacteroidota (0.04%) indicating the presence of contaminants of faecal sources.

High-throughput metagenomic assessment of Cango Cave microbiome-A South African limestone cave.

Microorganisms inhabiting caves exhibit medical or biotechnological promise, most of which have been attributed to factors such as antimicrobial activity or the induction of mineral precipitation. This dataset explored the shotgun metagenomic sequencing of the Cango cave microbial community in Oudtshoorn, South Africa. The aimed to elucidate both the structure and function of the microbial community linked to the cave. DNA sequencing was conducted using the Illumina NovaSeq platform, a next-generation sequencing. The data comprises 4,738,604 sequences, with a cumulative size of 1,180,744,252 base pairs and a GC content of 52%. Data derived from the metagenome sequences can be accessed through the bioproject number PRJNA982691 on NCBI. Using an online metagenome server, MG-RAST, the subsystem database revealed that bacteria displayed the highest taxonomical representation, constituting about 98.66%. Archaea accounted for 0.05%, Eukaryotes at 1.20%, viruses were 0.07%, while unclassified sequences had a representation of 0.02%. The most abundant phyla were Proteobacteria (81.74%), Bacteroidetes (10.57%), Actinobacteria (4.16%), Firmicutes (SK‒1.03%), Acidobacteria (0.20), and Planctomycetes (SK‒0.16%). Functional annotation using subsystem analysis revealed that clustering based on subsystems had 13.44%, while amino acids and derivatives comprised 11.41%. Carbohydrates sequences constituted 9.55%, along with other advantageous functional traits essential for growth promotion and plant management.

Novel Plasmodium falciparum k13 gene polymorphisms from Kisii County, Kenya during an era of artemisinin-based combination therapy deployment.

BACKGROUND: Currently, chemotherapy stands out as the major malaria intervention strategy, however, anti-malarial resistance may hamper global elimination programs. Artemisinin-based combination therapy (ACT) stands as the drug of choice for the treatment of Plasmodium falciparum malaria. Plasmodium falciparum kelch13 gene mutations are associated with artemisinin resistance. Thus, this study was aimed at evaluating the circulation of P. falciparum k13 gene polymorphisms from Kisii County, Kenya during an era of ACT deployment. METHODS: Participants suspected to have malaria were recruited. Plasmodium falciparum was confirmed using the microscopy method. Malaria-positive patients were treated with artemether-lumefantrine (AL). Blood from participants who tested positive for parasites after day 3 was kept on filter papers. DNA was extracted using chelex-suspension method. A nested polymerase chain reaction (PCR) was conducted and the second-round products were sequenced using the Sanger method. Sequenced products were analysed using DNAsp 5.10.01 software and then blasted on the NCBI for k13 propeller gene sequence identity using the Basic Local Alignment Search Tool (BLAST). To assess the selection pressure in P. falciparum parasite population, Tajima' D statistic and Fu & Li's D test in DnaSP software 5.10.01 was used. RESULTS: Out of 275 enrolled participants, 231 completed the follow-up schedule. 13 (5.6%) had parasites on day 28 hence characterized for recrudescence. Out of the 13 samples suspected of recrudescence, 5 (38%) samples were positively amplified as P. falciparum, with polymorphisms in the k13-propeller gene detected. Polymorphisms detected in this study includes R539T, N458T, R561H, N431S and A671V, respectively. The sequences have been deposited in NCBI with bio-project number PRJNA885380 and accession numbers SAMN31087434, SAMN31087433, SAMN31087432, SAMN31087431 and SAMN31087430 respectively. CONCLUSIONS: WHO validated polymorphisms in the k13-propeller gene previously reported to be associated with ACT resistance were not detected in the P. falciparum isolates from Kisii County, Kenya. However, some previously reported un-validated k13 resistant single nucleotide polymorphisms were reported in this study but with limited occurrences. The study has also reported new SNPs. More studies need to be carried out in the entire country to understand the association of reported mutations if any, with ACT resistance.

Trend of Malaria Burden Among Residents of Kisii County, Kenya After More Than a Decade Usage of Artemisinin Combined Therapies, 11-Year Laboratory Based Retrospective Study.

Background: Malaria remains a major vector borne disease globally, with the majority of the casualties reported in Africa. Despite this fact, there is drastic reduction in malaria infection using Artemisinin combined therapies (ACTs). Malaria is characterized by significant inconsistency in different geographical locations due to different confounding factors. There is need to identify zone-specific malaria trends and interventions to completely eliminate the disease. Thus the study was aimed at assessing the 11-year trend of microscopically confirmed malaria cases in Kisii County, Kenya, so as to devise area-specific evidence-based interventions, informed decisions, and to track the effectiveness of malaria control programs. Methods: This was a retrospective study carried out to determine 11-year malaria trend rates centered on the admission and laboratory records from health facilities located at four Sub-Counties in Kisii County, Kenya. Parasitological positivity rates of malaria were determined by comparing with the register records in health facilities which recorded confirmed malaria cases with the total number of monthly admissions over the entire year. Data was analyzed by using descriptive tools and chi-square test. Results: There were 36,946 suspect cases, with 8449 (22.8%) confirmed malaria cases reported in this study. The overall malaria slide positivity rate over the last 11 years in the study area was 22.6%. The months of April and August showed the largest number of malaria cases (63%). The age group of ≥18 years contained the most positive confirmed cases, having a prevalence rate of 2953 (35.45%). Out of the confirmed malaria cases, 2379 (28.1%) were males and 6070 (71.9%) were females The highest malaria prevalence rate was recorded in 2014, with Marani Sub-County recording the highest positivity rate of 37.94%. Conclusion: From the observed trends, malaria prevalence and transmission still remains stable in the study area. Thus more interventions need to be scaled up.

Limited Polymorphism in Plasmodium falciparum Artemisinin Resistance Kelch13-Propeller Gene Among Clinical Isolates from Bushenyi District, Uganda.

INTRODUCTION: Drug resistance remains a major challenge in malaria treatment, especially after the emergence of resistance to artemisinin-based combined therapies. Plasmodium falciparum Kelch13 gene mutations are implicated in conferring artemisinin resistance. Thus, this study was aimed at determining the occurrence of Kelch13 (K13) propeller resistance gene polymorphism mutations in Bushenyi district, Uganda. METHODS: Participants suspected to have malaria were recruited. P. falciparum was confirmed using antigen histidine-rich protein 2 (HRP2) (Pf) (Access Bio, Inc, USA) and microscopy. Malaria-positive patients were treated with artemeter-lumefantrine (AL). Blood was withdrawn from participants who tested positive for parasites after day 3 and kept in blood filter papers (ET31CHR; Whatman Limited, Kent, UK). DNA was extracted using chelex-suspension method. Nested polymerase chain reaction (PCR) was conducted and the second-round products sequenced using Sanger's method. Sequenced products were analyzed using DNAsp 5.10.01 software and then blasted on to the NCBI for K13-propeller gene sequence identity using the Basic Local Alignment Search Tool (BLAST). RESULTS: Out of 283 enrolled participants, 194 completed the follow-up schedule. A total of 134 (69%) had no parasites on day 3, while 60 (31%) had parasites on that day. Out of the 60 samples, 40 (62%) were positively amplified as P. falciparum, with polymorphisms in the K13-propeller gene detected in 3 (7.5%) out of the 40 amplicons. Polymorphisms at codon 1929, 1788 and 1801 were detected separately in one sample each. Sequences have been deposited in NCBI with accession numbers PRJNA720348 and PRJNA720800. CONCLUSION: Polymorphisms in the K13-propeller gene previously reported to be associated with artemisinin resistance were not detected in the P. falciparum isolates from Bushenyi district, Uganda. More studies need to be conducted on the new mutations detected so as to understand their association, if any, with ACT resistance.

Metagenomic Insight into the Community Structure of Maize-Rhizosphere Bacteria as Predicted by Different Environmental Factors and Their Functioning within Plant Proximity.

The rhizosphere microbiota contributes immensely to nutrient sequestration, productivity and plant growth. Several studies have suggested that environmental factors and high nutrient composition of plant's rhizosphere influence the structural diversity of proximal microorganisms. To verify this assertion, we compare the functional diversity of bacteria in maize rhizosphere and bulk soils using shotgun metagenomics and assess the influence of measured environmental variables on bacterial diversity. Our study showed that the bacterial community associated with each sampling site was distinct, with high community members shared among the samples. The bacterial community was dominated by Proteobacteria, Actinobacteria, Acidobacteria, Gemmatimonadetes, Bacteroidetes and Verrucomicrobia. In comparison, genera such as Gemmatimonas, Streptomyces, Conexibacter, Burkholderia, Bacillus, Gemmata, Mesorhizobium, Pseudomonas and Micromonospora were significantly (p ≤ 0.05) high in the rhizosphere soils compared to bulk soils. Diversity indices showed that the bacterial composition was significantly different across the sites. The forward selection of environmental factors predicted N-NO3 (p = 0.019) as the most influential factor controlling the variation in the bacterial community structure, while other factors such as pH (p = 1.00) and sulfate (p = 0.50) contributed insignificantly to the community structure of bacteria. Functional assessment of the sampling sites, considering important pathways viz. nitrogen metabolism, phosphorus metabolism, stress responses, and iron acquisition and metabolism could be represented as Ls > Rs > Rc > Lc. This revealed that functional hits are higher in the rhizosphere soil than their controls. Taken together, inference from this study shows that the sampling sites are hotspots for biotechnologically important microorganisms.

The diverse functional genes of maize rhizosphere microbiota assessed using shotgun metagenomics.

BACKGROUND: The geographical diversification in chemical, biological and physical properties of plant biospheres instigates heterogenicity in the proliferation of important soil microbiome. Controlling functions and structure of plant rhizosphere from a better understanding and prediction of a plant's immediate environment will help assess plant-microbe interplay, improve the productivity of plant ecosystems and improve plant response to adverse soil conditions. Here we characterized functional genes of the microbial community of maize rhizosphere using a culture-independent method. RESULTS: Our metadata showed microbial genes involved in nitrogen fixation, phosphate solubilization, quorum sensing molecules, trehalose, siderophore production, phenazine biosynthesis protein, daunorubicin resistance, acetoin, 1-aminocyclopropane-1-carboxylate deaminase, 4-hydroxybenzoate, disease control and stress-reducing genes (superoxidase dismutase, catalase, peroxidase, etc.). ß-Diversity showed that there is a highly significant difference between most of the genes mined from rhizosphere soil samples and surrounding soils. CONCLUSIONS: The high relative abundance of stress-reducing genes mined from this study showed that the sampling sites harbor not only important plant-beneficial organisms but also a hotspot for developing bio-fertilizers. Nevertheless, since most of these organisms are unculturable, mapping cultivation strategies for their growth could make them readily available as bio-inoculants and possible biotechnological applications in the future. © 2020 Society of Chemical Industry.

Shotgun Metagenomic Survey of Maize Soil Rhizobiome.

The plant soil rhizobiome induces critical functions in the plant proximal environment. Linkages between soil microbiota and primary functional attributes are underexplored. Here, we present the metagenomes of maize soil rhizosphere organisms with functional diversity associated with farms at two different municipalities in North West and Gauteng provinces of South Africa. We describe a plenteous and diverse microbial community.

In Vitro Antibacterial Activity of Crude Extracts of Artocarpus heterophyllus Seeds against Selected Diarrhoea-Causing Superbug Bacteria.

The current upsurge in resistance to conventional antibiotics, as well as high cost of orthodox medical treatment, called for the use of medicinal plants as an alternative therapy. This research was aimed at determining the antibacterial activity of Artocarpus heterophyllus seed extracts (Jackfruit as it is locally called) in the treatment of diarrhoea. Ethanolic and hexanolic seed crude extracts of the plant were screened for antidiarrhoeal activity against bacteria isolated from clinical samples (methicillin-resistant and susceptible Staphylococcus aureus, multidrug-resistant Pseudomonas aeruginosa, ciprofloxacin-resistant Salmonella typhimurium, and third-generation cephalosporin-resistant Escherichia coli). Plant phytochemical screening was conducted using standard methods. The antibacterial activity was carried out using the agar well diffusion method and compared to the standard antibiotics ceftriaxone and vancomycin. The minimum inhibitory concentration was determined by the microbroth dilution method, whereas the minimum bactericidal concentration was determined by plating out from microtitre plates with no visible growth. The results of phytochemical screening revealed the presence of tannins, flavonoids, reducing sugars, cardiac glycosides, saponins, and steroids from the prepared crude extracts. The ethanolic and hexanolic extracts had activity on multidrug-resistant Pseudomonas aeruginosa, methicillin-resistant Staphylococcus aureus, and methicillin-susceptible Staphylococcus aureus with the mean and standard error zone of inhibition that ranged from 8.5 ± 0.5 to 16.5 ± 0.25 mm; however, the extracts were found not to have activity on resistant E. coli and Salmonella typhimurium. The ethanolic crude extract had the lowest MIC and MBC values of 31.25 and 125 mg/ml, respectively, compared to the hexane extract which had the MIC and MBC values of 62.50 and 250 mg/ml, respectively. This provides the evidence for its usage as an alternative herbal remedy for the treatment of diarrhoea caused by susceptible and methicillin-resistant Staphylococcus aureus and multidrug resistant Pseudomonas aeruginosa.

Effect of storage on the quality of processed palm oil collected from local milling points within Ile-Ife, Osun State, Nigeria.

The influence of storage practices on physicochemical and microbial changes in crude palm oil (CPO) from milling points in Ile-Ife, Nigeria were investigated. Freshly milled CPO samples were collected from four traditional milling points, dispensed in 150 mL portions in sterile bottles and stored under two different conditions (sunlight reflection and in the dark, both at room temperature) for 4 months. Samples were obtained periodically during the storage period for microbiological and physicochemical analysis following established methods. The aerobic mesophilic (2.16 × 106 cfu/mL) and Enteric bacterial (1.6 × 106 cfu/mL) counts of the fresh CPO samples decreased during storage with those exposed to sunlight reflections having very high significant difference (P < 0.00) compared to those stored in the dark at P ≤ 0.05. The bacterial isolates were identified as Bacillus pasteurii (29%), Staphylococcus aureus (22%), Enterobacter aerogenes (17%), Micrococcus sp. (12%), Escherichia coli (8%), Pseudomonas aeruginosa (7%) and Serratia marcescens (5%). Of the physicochemical parameters studied, moisture content (MC) reduced significantly from between 2.55 and 5.50% in fresh sample to between 0.1 and 0.5% at the end of storage while the free fatty acids (FFA) increased from between 0.5 and 1.0% to between 2.2 and 3.1% respectively. Storage under the influence of sunlight resulted in significant increase in iodine value of CPO from Mills 1, 2 and 4, indicating oxidative instability of the palm oil. It could be concluded that storage of freshly milled palm oil at room temperature (in the dark or exposure to sunlight) for a period of 4 months resulted in reduced bacterial load, decrease in MC and stable peroxide value and FFA.

Prevalence of Dermatophytosis and Antifungal Activity of Ethanolic Crude Leaf Extract of Tetradenia riparia against Dermatophytes Isolated from Patients Attending Kampala International University Teaching Hospital, Uganda.

Dermatophyte infections are a global health problem but neglected in Uganda. This work aimed at determining prevalence of dermatophytosis and antifungal activity of ethanolic crude leaf extract of Tetradenia riparia against dermatophytes isolated from patients attending Kampala International University Teaching Hospital (KIU-TH), Uganda. A total of 100 samples of skin and nail scrapings were collected and processed using standard microscopy (KOH) and cultural methods. T. riparia leaves were collected and processed with 95% ethanol using standard extraction method. The crude leaves ethanolic extract was tested against three dermatophytes: Trichophyton tonsurans, T. mentagrophyte, and Microsporum audouinii using modified agar well diffusion method. Minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) of the ethanolic leaves crude extract were also determined using broth tube dilution and culture, respectively. Out of 100 samples collected, 49 (49%, 95%CI: 0.3930-0.5876) were found positive for microscopy. The prevalence of dermatophytosis was significantly (p=0.001) associated with age groups of participants with higher infection among those aged 11-20 and 21-30 years with 75.0% each. Out of the 49 that were positive by microscopy, 28 (57.15%, 95% CI: 0.1987-0.3739) were positive by culture. Thirty-one (31) fungal isolates were obtained which included both dermatophyte and non-dermatophyte fungi. T. verrucosum had highest distribution 6 (19.35%) among dermatophytes species while Aspergillus spp. were found to have highest distribution 7 (22.58%) among non-dermatophyte species. The result of the antidermatophytic test showed that T. riparia ethanolic crude leaves extract had activity against tested dermatophytes at 1 g/ml. MIC and MFC of the crude extract of T. riparia against tested dermatophytes ranged from 62.5 to 250 mg/ml and 125 to 500 mg/ml, respectively. The findings of this study reported the presence of dermatophytes causing dermatophytosis among patients attending KIU-TH. The results of the current study showed that T. riparia leaves ethanolic crude extract has antidermatophytic activity against tested dermatophytes.