Characterization and identification of long-chain hydrocarbon-degrading bacterial communities in long-term chronically polluted soil in Ogoniland: an integrated approach using culture-dependent and independent methods.

Escalating oil consumption has resulted in an increase in accidental spills of petroleum hydrocarbons, causing severe environmental degradation, notably in vulnerable regions like the Niger Delta. Complex mixture of these hydrocarbons particularly long-chain alkanes presents unique challenges in restoration of polluted environment due to their chemical properties. This study aimed to investigate the long-chain hydrocarbon-degrading bacterial communities within long-term chronically polluted soil in Ogoniland, by utilizing both traditional cultivation methods and modern culture-independent techniques. Results revealed that surface-polluted soil (SPS) and subsurface soil (SPSS) exhibit significantly higher total organic carbon (TOC) ranging from 5.64 to 5.06% and total petroleum hydrocarbons (TPH) levels ranging from 36,775 ppm to 14,087 ppm, compared to unpolluted soil (UPS) with 1.97% TOC and 479 ppm TPH, respectively. Analysis of carbon chain lengths reveals the prevalence of longer-chain alkanes (C20-28) in the surface soil. Culture-dependent methods, utilizing crude oil enrichment (COE) and paraffin wax enrichment (PWE), yield 47 bacterial isolates subjected to a long-chain alkane degradation assay. Twelve bacterial strains demonstrate significant degradation abilities across all enriched media. Three bacterial members, namely Pseudomonas sp. (almA), Marinomonas sp. (almA), and Alteromonas (ladA), exhibit genes responsible for long-chain alkane degradation, demonstrating efficiency between 50 and 80%. Culture-independent analysis reveals that surface SPS samples exhibit greater species richness and diversity compared to subsurface SPSS samples. Proteobacteria dominates as the phylum in both soil sample types, ranging from 22.23 to 82.61%, with Firmicutes (0.2-2.22%), Actinobacteria (0.4-3.02%), and Acidobacteria (0.1-3.53%) also prevalent. Bacterial profiles at genus level revealed that distinct variations among bacterial populations between SPS and SPSS samples comprising number of hydrocarbon degraders and the functional predictions also highlight the presence of potential catabolic genes (nahAa, adh2, and cpnA) in the polluted soil. However, culture-dependent analysis only captured a few of the dominant members found in culture-independent analysis, implying that more specialized media or environments are needed to isolate more bacterial members. The findings from this study contribute valuable information to ecological and biotechnological aspects, aiding in the development of more effective bioremediation applications for restoring oil-contaminated environments.

Human viral pathogens in the wastewater-source water-drinking water continuum: Evidence, health risks, and lessons for future outbreaks in low-income settings.

Human viral pathogens, including SARS-CoV-2 continue to attract public and research attention due to their disruption of society, global health, and the economy. Several earlier reviews have investigated the occurrence and fate of SARS-CoV-2 in wastewater, and the potential to use such data in wastewater-based epidemiology. However, comprehensive reviews tracking SARS-CoV-2 and other viral pathogens in the wastewater-water-drinking water continuum and the associated risk assessment are still lacking. Therefore, to address this gap, the present paper makes the following contributions: (1) critically examines the early empirical results to highlight the occurrence and stability of SARS-CoV-2 in the wastewater-source water-drinking water continuum, (2) discusses the anthropogenic and hydro(geo)logical processes controlling the circulation of SARS-CoV-2 in the wastewater-source water-drinking water continuum, (3) discusses the risky behaviour, drivers and high-risk settings in the wastewater-source water-drinking water continuum, (4) uses the available empirical data on SARS-CoV-2 occurrence in the wastewater-source water-drinking water continuum to discuss human health risks from multiple exposure pathways, gendered aspects of SARS-CoV-2 transmission via shared on-site sanitation systems, and (5) develops and risk mitigation strategy based on the available empirical evidence and quantitative human risk assessment data. Finally, it presents a comprehensive research agenda on SARS-CoV-2/COVID-19 to guide the mitigation of future similar outbreaks in low-income settings.

High-throughput amplicon sequencing datasets of microbial community in soils irrigated by quicklime and fly ash-treated acid mine drainage water.

In water-stressed regions, the use of treated acid mine drainage (AMD) water for irrigated agriculture has been suggested as an alternative to address the shortage of fresh water sources. However, the short and long-term impact of using such (un)treated AMD water on soil health, particularly the microbiome structure and functional capacity, is not known. We present high-throughput amplicon sequence (HTS) datasets of purified microbial metacommunity DNA of soils under Irish potato production irrigated by quicklime and fly ash treated AMD water. The irrigation treatments included quicklime treated AMD water (A1Q and A2Q; n = 16), and quicklime and fly ash-treated AMD water (AFQ; n = 5), untreated AMD water (uAMD; n = 7) and control group using tap water (n = 5). The V1-V3 hypervariable region of the 16S rRNA gene from each sample were sequenced on an Illumina MiSeq to generate these HTS datasets. The raw sequences underwent quality-checking, demultiplexing into FASTQ files, and processing using MOTHUR pipeline (v1.40.0). Th quality reads classified into taxonomic ids (phylum, class, order, family, and genus) using the Naïve Bayesian classifier algorithm against the SILVA database (v132) and were assigned to operational taxonomic units (OTUs) based on the pairwise distance matrix (Euclidean distance matrix). The applicability of the HTS datasets was confirmed by microbial taxa at the phylum level. All HTS datasets are available through the BioSample Submission Portal under the BioProject ID PRJNA974836 (https://www.ncbi.nlm.nih.gov/bioproject/974836).

Beach sand mycobiome: The silent threat of pathogenic fungi and toxic metal contamination for beachgoers.

Emphasis is always placed on bacterial but not fungal pathogens in marine environments. We analysed the fungal diversity, functional predictions, and toxic metals and metalloids contamination in beach sand from different South African locations. Results revealed a diverse fungal community, with Ascomycota, Rozellomycota, and Basidiomycota being the dominant phyla. Functional predictions highlighted fungal metabolic pathways related to of carbohydrates, amino acids, and lipids, in different beach samples. Elevated concentrations of toxic metals and metalloids were detected in Central and Harbour beach sands, likely due to anthropogenic activities. Correlations among different elements were observed, suggesting complex interactions in the coastal environment. Fungal pathogens like Cladosporium, Fusarium, Aspergillus, and Candida in beach sands raise potential public health risk concerns. Therefore, monitoring fungal diversity (including pathogens) alongside bacterial contamination in beach environments is imperative. The results contribute to understanding fungal community dynamics, functional potential, toxic metal and metalloid contamination, and potential risks associated with beach sand ecosystems.

Bioaccumulation, Bioindication and Health Risk Assessment of Heavy Metals in Cape Horse Mackerel (Trachurus trachurus) and Slinger Seabream (Chrysoblephus puniceus) in the Durban Basin and Cape Vidal, South Africa.

The bioaccumulation of heavy metals (HMs) in marine fish is a growing global concern due to potential human health risks. The study analyzed HM in the muscle tissue, gills, and gut of adult male and female cape horse mackerel and slinger seabream caught in the polluted Durban Basin and pristine Cape Vidal from April 2018 to February 2019. Results revealed interspecific, spatial, and organ-specific variability in HM levels. In the Durban Basin, slinger seabream had bioaccumulation (in mg/kg) of As (2.3 ± 0.2), Cr (2.6 ± 0.2), Ni (2.0 ± 0.1), and Pb (4.1 ± 0.3) while cape horse mackerel had Ni (1.6 ± 0.2), Pb (4.7 ± 0.6), and Zn (52 ± 3.01) exceeding World Health Organization (WHO) regulatory limits. Metal pollution index (MPI) values were also higher in Durban Basin (> 5.13) than Cape Vidal (< 3.32) for both species' muscles. Liver and gills of slinger seabream and gut of cape horse mackerel exhibited higher HM accumulation patterns proportionate to the environmental concentrations, indicating the bioindicative potential of HM pollution by the two species. Risk assessment indicated that both fish species had target hazard quotient > 1 for Cr, and target cancer risk < 10-4 for Pb, implying significant potential non-carcinogenic and carcinogenic health risks associated with fish consumption from the Durban Basin. The study recommends daily consumption limits of 16 g/day for slinger seabream and 14 g/day for cape horse mackerel to ensure health safety. The findings contribute to the understanding of HM pollution in the Durban Basin and provide important information for decision-makers and policymakers in developing effective strategies to mitigate and manage HM contamination in fish populations.

Untapped talents: insight into the ecological significance of methanotrophs and its prospects.

The deep ocean is a rich reservoir of unique organisms with great potential for bioprospecting, ecosystem services, and the discovery of novel materials. These organisms thrive in harsh environments characterized by high hydrostatic pressure, low temperature, and limited nutrients. Hydrothermal vents and cold seeps, prominent features of the deep ocean, provide a habitat for microorganisms involved in the production and filtration of methane, a potent greenhouse gas. Methanotrophs, comprising archaea and bacteria, play a crucial role in these processes. This review examines the intricate relationship between the roles, responses, and niche specialization of methanotrophs in the deep ocean ecosystem. Our findings reveal that different types of methanotrophs dominate specific zones depending on prevailing conditions. Type I methanotrophs thrive in oxygen-rich zones, while Type II methanotrophs display adaptability to diverse conditions. Verrumicrobiota and NC10 flourish in hypoxic and extreme environments. In addition to their essential role in methane regulation, methanotrophs contribute to various ecosystem functions. They participate in the degradation of foreign compounds and play a crucial role in cycling biogeochemical elements like metals, sulfur, and nitrogen. Methanotrophs also serve as a significant energy source for the oceanic food chain and drive chemosynthesis in the deep ocean. Moreover, their presence offers promising prospects for biotechnological applications, including the production of valuable compounds such as polyhydroxyalkanoates, methanobactin, exopolysaccharides, ecotines, methanol, putrescine, and biofuels. In conclusion, this review highlights the multifaceted roles of methanotrophs in the deep ocean ecosystem, underscoring their ecological significance and their potential for advancements in biotechnology. A comprehensive understanding of their niche specialization and responses will contribute to harnessing their full potential in various domains.

Medium-chain alkane biodegradation and its link to some unifying attributes of alkB genes diversity.

Hydrocarbon footprints in the environment, via biosynthesis, natural seepage, anthropogenic activities and accidents, affect the ecosystem and induce a shift in the healthy biogeochemical equilibrium that drives needed ecological services. In addition, these imbalances cause human diseases and reduce animal and microorganism diversity. Microbial bioremediation, which capitalizes on functional genes, is a sustainable mitigation option for cleaning hydrocarbon-impacted environments. This review focuses on the bacterial alkB functional gene, which codes for a non-heme di­iron monooxygenase (AlkB) with a di­iron active site that catalyzes C8-C16 medium-chain alkane metabolism. These enzymes are ubiquitous and share common attributes such as being controlled by global transcriptional regulators, being a component of most super hydrocarbon degraders, and their distributions linked to horizontal gene transfer (HGT) events. The phylogenetic approach used in the HGT detection suggests that AlkB tree topology clusters bacteria functionally and that a preferential gradient dictates gene distribution. The alkB gene also acts as a biomarker for bioremediation, although it is found in pristine environments and absent in some hydrocarbon degraders. For instance, a quantitative molecular method has failed to link alkB copy number to contamination concentration levels. This limitation may be due to AlkB homologues, which have other functions besides n-alkane assimilation. Thus, this review, which focuses on Pseudomonas putida GPo1 alkB, shows that AlkB proteins are diverse but have some unifying trends around hydrocarbon-degrading bacteria; it is erroneous to rely on alkB detection alone as a monitoring parameter for hydrocarbon degradation, alkB gene distribution are preferentially distributed among bacteria, and the plausible explanation for AlkB affiliation to broad-spectrum metabolism of hydrocarbons in super-degraders hitherto reported. Overall, this review provides a broad perspective of the ecology of alkB-carrying bacteria and their directed biodegradation pathways.

Wnt Signaling Pathway Collapse upon ß-Catenin Destruction by a Novel Antimicrobial Peptide SKACP003: Unveiling the Molecular Mechanism and Genetic Activities Using Breast Cancer Cell Lines.

Despite progress in breast cancer treatment, the survival rate for patients with metastatic breast cancer remains low due to chemotherapeutic agent resistance and the lack of specificity of the current generation of cancer drugs. Our previous findings indicated that the antimicrobial peptide SKACP003 exhibited anticancer properties, particularly against the MCF-7, MDA-MB-231, and MDA-MB-453 breast cancer cell lines. However, the mechanism of SKACP003-induced cancer cell death is unknown. Here, we investigated the molecular mechanism by which SKACP003 inhibits the cell cycle, cell proliferation, and angiogenesis in breast cancer cell lines. The results revealed that all the breast cancer cell lines treated at their IC50 values significantly inhibited the replicative phase of the cell cycle. The SKACP003-induced growth inhibition induced apoptosis, as evidenced by a decrease in BCL-2 and an increase in BAX and caspase gene (Cas-3, Cas-8, and Cas-9) expression. Reduced expression of the ß-Catenin signaling pathway was associated with the SKACP003-induced apoptosis. SKACP003-treated breast cancer cells showed decreased expression of Wnt/ß-Catenin targeting genes such as C-Myc, P68, and COX-2 and significant downregulation of CDK-4 and CDK-6 genes. Furthermore, cytoplasmic ß-catenin protein levels in SKACP003-treated cell lines were significantly lower than in control cell lines. The results of the current study suggest that the newly identified antimicrobial peptide SKACP003 has great potential as a candidate for specifically targeting the ß-catenin and thus significantly reducing the progression and prognosis of breast cancer cell lines.

Unravelling the genetic and functional diversity of dominant bacterial communities involved in manure co-composting bioremediation of complex crude oil waste sludge.

The present study aimed to characterize the bacterial community and functional diversity in co-composting microcosms of crude oil waste sludge amended with different animal manures, and to evaluate the scope for biostimulation based in situ bioremediation. Gas chromatography-mass spectrometry (GC-MS) analyses revealed enhanced attenuation (>90%) of the total polyaromatic hydrocarbons (PAHs); the manure amendments significantly enhancing (up to 30%) the degradation of high molecular weight (HMW) PAHs. Microbial community analysis showed the dominance (>99% of total sequences) of sequences affiliated to phyla Proteobacteria, Firmicutes, Actinobacteria and Bacteroidetes. The core genera enriched were related to hydrocarbon metabolism (Pseudomonas, Delftia, Methylobacterium, Dietzia, Bacillus, Propionibacterium, Bradyrhizobium, Streptomyces, Achromobacter, Microbacterium and Sphingomonas). However, manure-treated samples exhibited high number and heterogeneity of unique operational taxonomic units (OTUs) with enrichment of additional hydrocarbon-degrading bacterial taxa (Proteiniphilum, unclassified Micrococcales, unclassified Lachnospiraceae, Sphingobium and Stenotrophomonas). Thirty-three culturable hydrocarbon-degrading microbes were isolated from the co-composting microcosms and mainly classified into Burkholderia, Sanguibacter, Pseudomonas, Bacillus, Rhodococcus, Lysinibacillus, Microbacterium, Brevibacterium, Geobacillus, Micrococcus, Arthrobacter, Cellulimicrobacterium, Streptomyces Dietzia,etc,. that was additionally affirmed with the presence of catechol 2,3-dioxygenase gene. Finally, enhanced in situ degradation of total (49%), LMW (>75%) and HMW PAHs (>35%) was achieved with an enriched bacterial consortium of these microbes. Overall, these findings suggests that co-composting treatment of crude oil sludge with animal manures selects for intrinsically diverse bacterial community, that could be a driving force behind accelerated bioremediation, and can be exploited for engineered remediation processes.

Evolution and Emergence of Antibiotic Resistance in Given Ecosystems: Possible Strategies for Addressing the Challenge of Antibiotic Resistance.

Antibiotics were once considered the magic bullet for all human infections. However, their success was short-lived, and today, microorganisms have become resistant to almost all known antimicrobials. The most recent decade of the 20th and the beginning of the 21st century have witnessed the emergence and spread of antibiotic resistance (ABR) in different pathogenic microorganisms worldwide. Therefore, this narrative review examined the history of antibiotics and the ecological roles of antibiotics, and their resistance. The evolution of bacterial antibiotic resistance in different environments, including aquatic and terrestrial ecosystems, and modern tools used for the identification were addressed. Finally, the review addressed the ecotoxicological impact of antibiotic-resistant bacteria and public health concerns and concluded with possible strategies for addressing the ABR challenge. The information provided in this review will enhance our understanding of ABR and its implications for human, animal, and environmental health. Understanding the environmental dimension will also strengthen the need to prevent pollution as the factors influencing ABR in this setting are more than just antibiotics but involve others like heavy metals and biocides, usually not considered when studying ABR.

Correlations Between Root Metabolomics and Bacterial Community Structures in the Phragmites australis Under Acid Mine Drainage-Polluted Wetland Ecosystem.

Despite root microecology playing critical role in plant growth and fidelity, relatively few studies have focused on the link between the microbial communities and root metabolome in the aquatic macrophytes under heavy metal (HM) pollution. Using high-throughput metagenomic sequencing, targeted metabolomics and community-level physiological profile analyses, we investigated the symbiotic associations between Phragmites australis with rhizospheric bacterial communities under differing acid mine drainage (AMD) pollution. Results indicated that AMD pollution and root localization significantly affected root metabolome profiles. Higher accumulation of adenosine monophosphate, inosine, methionine, carnitine and dimethylglycine were observed in the rhizosphere under AMD than non-AMD habitat. Overall, the bacterial diversity and richness, and functional (metabolic) diversity were lower under high-AMD pollution. While non-AMD site was enriched with members of phylum Firmicutes, Proteobacteria were the most abundant taxa in the rhizosphere and endosphere under AMD-polluted sites. Further, plant growth promoting rhizobacteria (Rhizobium, Delftia, Bradyrhizobium, and Mesorhizobium) and metal-tolerant bacteria (Bacillus, Arthrobacter, Massilia and Methylocystis) were most abundant in AMD-polluted than non-AMD habitat. Finally, pH, TDS (total dissolved solids), Cu, Cr, Fe, and Zn content were the key environmental factors that strongly contributed to the spatial perturbation of rhizospheric metabolites, proteobacterial and acidobacterial taxa. Overall, the study linked the differential endospheric and rhizospheric bacterial community and metabolite profiles in P. australis under AMD environment and provided insights into HM adaptability and phytoremediation potential.

Taxonomic and Functional Distribution of Bacterial Communities in Domestic and Hospital Wastewater System: Implications for Public and Environmental Health.

The discharge of untreated hospital and domestic wastewater into receiving water bodies is still a prevalent practice in developing countries. Unfortunately, because of an ever-increasing population of people who are perennially under medication, these wastewaters contain residues of antibiotics and other antimicrobials as well as microbial shedding, the direct and indirect effects of which include the dissemination of antibiotic resistance genes and an increase in the evolution of antibiotic-resistant bacteria that pose a threat to public and environmental health. This study assessed the taxonomic and functional profiles of bacterial communities, as well as the antibiotic concentrations in untreated domestic wastewater (DWW) and hospital wastewater (HWW), using high-throughput sequencing analysis and solid-phase extraction coupled to Ultra-high-performance liquid chromatography Mass Spectrometry (UHPLC-MS/MS) analysis, respectively. The physicochemical qualities of both wastewater systems were also determined. The mean concentration of antibiotics and the concentrations of Cl-, F- and PO43 were higher in HWW samples than in DWW samples. The phylum Firmicutes was dominant in DWW with a sequence coverage of 59.61% while Proteobacteria was dominant in HWW samples with a sequence coverage of 86.32%. At genus level, the genus Exiguobacterium (20.65%) and Roseomonas (67.41%) were predominant in DWW and HWW samples, respectively. Several pathogenic or opportunistic bacterial genera were detected in HWW (Enterococcus, Pseudomonas and Vibrio) and DWW (Clostridium, Klebsiella, Corynebacterium, Bordetella, Staphylocccus and Rhodococcus) samples. Functional prediction analysis indicated the presence of beta-lactam resistance, cationic antimicrobial peptide (CAMP) resistance and vancomycin resistance genes in HWW samples. The presence of these antibiotic resistance genes and cassettes were positively correlated with the presence of pathogens. These findings show the risk posed to public and environmental health by the discharge of untreated domestic and hospital wastewaters into environmental water bodies.

The occurrence of opioid compounds in wastewater treatment plants and their receiving water bodies in Gauteng province, South Africa.

The continuous influx of opioid compounds into aquatic environments has become an increasing and persistent concern, due to their extensive use. This is especially alarming as wastewater treatment plants (WWTPs) are unable to completely remove them. Despite the reported health concerns, the occurrence of opioid compounds in the environment has not received much attention. The present study investigates the occurrence of 19 opioids in four WWTPs and their respective receiving water bodies. All wastewater samples revealed opioids at concentration ranging from ng/L to µg/L with most influents having higher concentrations than effluents. WWTPs appeared to perform poorly (p > 0.05 between influents and effluents), and were unable to remove some opioids including Methadone (-27.3%) from the Leeuwkuil WWTP, Codeine (-21.7%) and Thebaine (-3.77%) from the Sandspruit WWTP, and Hydrocodone (-1.06%) from the Meyerton WWTP, respectively. Samples collected from the Leeuwkuil WWTP were the most contaminated, with eighteen out of nineteen opioid analogues exceeding 1 µg/L. Upstream surface water contained less opioids (most < LOQ) than downstream (p < 0.05), with Hydrocodone, Oxycodone, Hydromorphone, Fentanyl, Ketamine and Dihydrocodeine not detected. The occurrence of high concentrations of opioid analogues in downstream surface water (298 ng/L -10.8 µg/L for Klip River, 4.49 ng/L -13.1 µg/L for Vaal River, 70.5 ng/L -10.0 µg/L for Soutspruit River and 8.0 ng/L - 2.43 µg/L for Sun Spruit River) was directly linked to their mass loads in the respective wastewater effluent samples.

Local Geomorphological Gradients and Land Use Patterns Play Key Role on the Soil Bacterial Community Diversity and Dynamics in the Highly Endemic Indigenous Afrotemperate Coastal Scarp Forest Biome.

Southern Afrotemperate forests are small multi-layered and highly fragmented biodiversity rich biomes that support unique flora and fauna endemism. However, little is known about the microbial community and their contribution to these ecosystems. In this study, high throughput sequencing analysis was used to investigate the soil bacterial community structure and function, and understand the effect of local topography/geomorphological formations and land use patterns on a coastal scarp forest. Soil samples were collected from three forest topography sites: upper (steeper gradients, 30-55°; open canopy cover, <30%), mid (less steep, 15-30°; continuous forest canopy, >80%), and lower (flatter gradient, <15°; open canopy cover, 20-65%), and from the adjacent sugarcane farms. Results indicated that forest soils were dominated by members of phyla Proteobacteria (mainly members of α-proteobacteria), Actinobacteria, Acidobacteria, Firmicutes, and Planctomycetes, while Actinobacteria and to a lesser extent ß-proteobacteria and γ-proteobacteria dominated SC soils. The core bacterial community clustered by habitat (forest vs. sugarcane farm) and differed significantly between the forest topography sites. The Rhizobiales (genera Variibacter, Bradyrhizobium, and unclassified Rhizobiales) and Rhodospirallales (unclassified Rhodospirillum DA111) were more abundant in forest mid and lower topographies. Steeper forest topography (forest_upper) characterized by the highly leached sandy/stony acidic soils, low in organic nutrients (C and N) and plant densities correlated to significant reduction of bacterial diversity and richness, associating significantly with members of order Burkholderiales (Burkholderia-Paraburkholderia, Delftia, and Massilia) as the key indicator taxa. In contrast, changes in the total nitrogen (TN), soil organic matter (SOM), and high acidity (low pH) significantly influenced bacterial community structure in sugarcane farm soils, with genus Acidothermus (Frankiales) and uncultured Solirubrobacterales YNFP111 were the most abundant indicator taxa. Availability of soil nutrients (TN and SOM) was the strongest driver of metabolic functions related to C fixation and metabolism, N and S cycling; these processes being significantly abundant in forest than sugarcane farm soils. Overall, these results revealed that the local topographical/geomorphological gradients and sugarcane farming affect both soil characteristics and forest vegetation (canopy coverage), that indirectly drives the structure and composition of bacterial communities in scarp forest soils.

Influences of geochemical factors and substrate availability on Gram-positive and Gram-negative bacterial distribution and bio-processes in ageing municipal landfills.

Bacteria are primary agents of organic substrate metabolisation and elemental cycling in landfills. Two major bacterial groups, namely, Gram-positive (GP) and Gram-negative (GN), drive independent metabolic functions that contribute to waste stabilisation. There is a lack of explicit exploration of how these different bacterial guilds respond to changing carbon (C) availability and substrate depletion as landfills age and how landfill geochemistry regulates their distribution. This study investigated and compared the abundance and vertical distribution of GP and GN bacteria in 14- and 36-year-old municipal landfills and explored linkages among bacterial groups, nutrient elements, heavy metals and soil texture. We found higher GP bacteria in the 14-year-old landfill, while GN bacteria dominated the 36-year-old landfill. The non-metric multidimensional scaling (nMDS) analysis showed that dissimilarities in the relative abundance of the GP and GN bacteria were linked distinctly to landfill age, and not depth. In support of this inference, we further found that GP and GN bacteria were negatively correlated with heavy metals and essential nutrients in the 14- and 36-year-old landfills, respectively. Notably, the GP/GN ratio, an indicator of relative C available for bacterial mineralisation, was greater in the14-year-old landfill, suggesting greater C availability. Conversely, the C to N ratio was higher in the 36-year-old landfill, indicating lower N mineralisation. Collectively, the results of the study reveal key insights into how landfill ageing and stabilisation influence distinct functional shifts in the abundance of GP and GN bacteria, and these are mainly driven by changes in C and N bioavailability.

Fungal and metabolome diversity of the rhizosphere and endosphere of Phragmites australis in an AMD-polluted environment.

Symbiotic associations with rhizospheric microbial communities coupled with the production of metabolites are key adaptive mechanisms by metallophytes to overcome metal stress. However, little is known about pseudometallophyte Phragmites australis interactions with fungal community despite commonly being applied in wetland phytoremediation of acid mine drainage (AMD). In this study, fungal community diversity and metabolomes production by rhizosphere and root endosphere of P. australis growing under three different AMD pollution gradient were analyzed. Our results highlight the following: 1) Ascomycota and Basidiomycota were dominant phyla, but the diversity and richness of taxa were lower within AMD sites with Penicillium, Candida, Saccharomycetales, Vishniacozyma, Trichoderma, Didymellaceae, and Cladosporium being enriched in the root endosphere and rhizosphere in AMD sites than non-AMD site; 2) non-metric multidimensional scaling (NMDS) of 73 metabolomes revealed spatially defined metabolite exudation by distinct root parts (rhizosphere vs endosphere) rather than AMD sites, with significant variability occurring within the rhizosphere correlating to pH, TDS, Fe, Cr, Cu and Zn content changes; 3) canonical correspondence analysis (CCA) confirmed specific rhizospheric fungal taxonomic changes are driven by pH, TDS, heavy metals, and stress-related metabolomes produced. This is the first report that gives a snapshot on the complex endophytic and rhizospheric fungal community structure and metabolites perturbations that may be key in the adaptability and metal phytoremediation by P. australis under AMD environment.

Shifts in Bacterial Diversity During the Spontaneous Fermentation of Maize Meal as Revealed by Targeted Amplicon Sequencing.

Maize meal was allowed to undergo uncontrolled fermentation in the laboratory, in simulation of the traditional method of fermentation as practised in most African households. During the fermentation process, samples were collected daily for 11 days. Physico-chemical analysis of the fermenting slurry and metagenomics analysis of the microbial community using 16S rRNA demonstrated an interrelationship between the changes in the properties of the fermentation environment and the successional interplay of the microbial community. The first 24 h of fermentation at pH of 6.5 was characterised by the proliferation of probiotic Lactobacillus and Bifidobacterium, with their relative abundance being 40.7% and 29.9%, respectively. However, prolonged fermentation and a drop in pH from 5.3 to 3.7 caused a decline and finally an absence of these probiotic bacteria which were replaced by Clostridium spp. with a relative abundance of between 97% and 99% from day 5 to day 11. This study demonstrated that prolonged fermentation of maize meal is not ideally suited for the proliferation of probiotic nutritionally beneficial bacteria.

Distribution and comparison of bacterial communities in HVAC systems of two university buildings: Implications for indoor air quality and public health.

The installation of HVAC systems in building is meant to enhance indoor air quality as well as increase comfort to occupants. However, HVAC systems have also become a vehicle of contamination of indoor air with potentially pathogenic microorganisms. DNA was extracted from ten HVAC filter dust samples collected from two buildings and subjected to high throughput sequencing analysis to determine the bacterial community structure. Further, the Phylogenetic Investigation of Communities by Reconstruction of Unobserved States (PICRUSt2) software was used to predict the potential functional capabilities of the bacterial communities. Sequencing analysis led to the identification of five major bacterial phyla, including Proteobacteria, Cyanobacteria, Actinobacteria, Firmicutes and Bacteroidetes. At genus level, Mycobacterium, Bacillus, Cupriavidus, Hyphomicrobium and Mesorhizobium were the most dominant. With the exception of the later two bacterial genera, the first three are potential pathogens whose presence in HVAC systems poses a significant public health risk, especially among immunocompromised individuals. Nine pathways associated with antibiotics resistance and bacterial pathogenicity were identified, including polymyxin resistance and peptidoglycan biosynthesis pathways. Further, investigation of the relationship between the detected bacterial meta-communities and predicted potential virulence factors (antibiotic resistance and pathogenic genes) led to the detection of 350 positive associations among 43 core bacteria, 2 pathogenic genes (sitA and uidA) and 14 resistance genes. Overall, the heterogeneous nature of microorganisms found in HVAC systems observed in this study shows that HVAC systems are the origin of airborne infections in indoor environments, and must be periodically cleaned and disinfected to avoid the build-up of pathogens, and the subsequent exposure of human occupants of these pathogens.

Azole antifungal resistance in fungal isolates from wastewater treatment plant effluents.

Wastewater treatment plants (WWTPs) can be significant sources of antifungal resistant fungi, which can disseminate further in the environment by getting into rivers together with effluents discharged from WWTPs and pose a risk for human health. In this study, the presence of azole resistance was determined in fungal isolates from treated effluents of two WWTPs using the standard microdilution method from Clinical and Laboratory Standards Institute (CLSI). A total of 41 fungal isolates representing 23 fungal species and 16 fungal genera were obtained. Fungal genera related to the known human and/or plant pathogens such as Aspergillus, Fusarium, and Candida were detected. Among the observed species, the susceptibility of Aspergillus fumigatus and Fusarium oxysporum was tested against fluconazole (FCZ), ketoconazole (KTZ), itraconazole (ITZ), and voriconazole (VCZ). The isolate A. fumigatus was susceptible to KTZ, ITZ, and VCZ, while it showed resistance against FCZ. On the contrast, the isolate F. oxysporum showed resistance to KTZ, ITZ, and VCZ. Comparatively, VCZ showed highest activity against both A. fumigatus and F. oxysporum. Analysis of the gene Cyp51A for the A. fumigatus isolate showed no evidence of drug resistance that could be related to point mutations and/or tandem repeats in the gene. To the best of our knowledge, this is the first susceptibility test study on A. fumigatus and F. oxysporum isolates from the WWTPs of South Africa. In conclusion, this study indicated an urgent need for thorough investigation with larger group of fungal isolates from different regions of South Africa to broadly understand the role of WWTPs in the dissemination of azole antifungal drug resistance.

Community diversity metrics, interactions, and metabolic functions of bacteria associated with municipal solid waste landfills at different maturation stages.

Municipal landfills are hot spots of dynamic bioprocesses facilitated by complex interactions of a multifaceted microbiome, whose functioning in municipal landfills at different maturing stages is poorly understood. This study determined bacterial community composition, interaction conetworks, metabolic functions, and controlling physicochemical properties in two landfills aged 14 and 36 years. High throughput sequencing revealed a similar distribution of bacterial diversity, evenness, and richness in the 14- and 36-year-old landfills in the 0-90 cm depth. At deeper layers (120-150 cm), the 14-year-old landfill had significantly greater bacterial diversity and richness indicating that it is a more active microcosm than the 36-year-old landfill, where phylum Epsilonbacteraeota was overwhelmingly dominant. The taxonomic and functional diversity in the 14-year-old landfill was further reflected by the abundant presence of indicator genera Pseudomonas,Lutispora,Hydrogenspora, and Sulfurimonas coupled with the presence of biomarker enzymes associated with carbon (C), nitrogen (N), and sulfur (S) metabolism. Furthermore, canonical correspondence analysis revealed that bacteria in the 14-year-old landfill were positively correlated with high C, N, S, and phosphorus resulting in positive cooccurrence interactions. In the 36-year-old landfill, negative coexclusion interactions populated by members of N fixing Rhizobiales were dominant, with metabolic functions and biomarker enzymes predicting significant N fixation that, as indicated by interaction network, potentially inhibited ammonia-intolerant bacteria. Overall, our findings show that diverse bacterial community in the 14-year-old landfill was dominated by copiotrophs associated with positive conetworks, whereas the 36-year-old landfill was dominated by lithotrophs linked to coexclusion interactions that greatly reduced bacterial diversity and richness.