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The superfamily Tenebrionoidea is one of the most challenging clades in the beetle tree-of-life owing to its vast species richness and complex taxonomic history. Within this group, the family Melandryidae has long been overlooked and its systematics remains poorly known. Using available sequence data, we infer the most comprehensive phylogeny of Melandryidae to date. Our results support the polyphyly of Melandryidae with three independent clades spread across Tenebrionoidea. To accommodate these results, we restrict the status of Melandryidae and resurrect the family Osphyidae stat. rev. The third clade corresponding to the tribe Serropalpini pro parte is placed as incertae sedis within Tenebrionoidea pending further investigation and additional taxon sampling to resolve its phylogenetic placement.

Over the past two decades, research on the conceptualization, measurement, and refinement of stigma has grown extensively due to increasing recognition of stigma as a driver of adverse health outcomes. While these lines of research have generally recognized that stigma is enacted in social relationships, few analysts have characterized how the structure and composition of these relationships influence stigma, particularly among immigrant populations. In response, this paper integrates data from social network analysis and in-depth interviews to explore the experiences of and responses to stigma by African immigrants living with HIV in the United States.All participants reported that they anticipated, experienced, and internalized stigma within their personal networks. Many concealed their status and disclosed to only trusted associates, family members, and medical providers. Building on findings from previous studies, we found that the meanings and belief systems (particularly African cultural discourses that link HIV with mortality, immorality, retribution, and silence) matter for how stigma is assigned, enacted, experienced, and resisted. Our analyses also revealed that the structure of participants personal networks (i.e, the extent to which their associates were connected to each other, and how integrated/involved they were in these network relationships) intensified or diluted their exposure to stigmatizing discourses, depending on the composition (resourcefulness/quality) of their personal networks. Such network connections (i.e., social capital) translate into rewards if they are supportive and accepting, and costs if they stigmatize.By showing how individuals can use their social networks to stigmatize or support their peers, this study advances theoretical expositions on (1) how the meanings and belief systems held by individuals matter for understanding social network/structural processes, (2) how social networks shape how stigma is assigned, enacted, experienced and resisted,(3) the costs and downsides of social capital, which are often neglected through emphasis on its salutary impacts. Our findings underscore the need for interventions that leverage the salient properties of personal networks to understand, conceptualize, measure, and reduce stigma.

Abstract Tobacco use significantly influences the oral microbiome. However, less is known about how different tobacco products specifically impact the oral microbiome over time. To address this knowledge gap, we characterized the oral microbiome of cigarette users, smokeless tobacco users, and non-users over 4 months (four time points). Buccal swab and saliva samples (n = 611) were collected from 85 participants. DNA was extracted from all samples and sequencing was carried out on an Illumina MiSeq, targeting the V3–V4 region of the 16S rRNA gene. Cigarette and smokeless tobacco users had more diverse oral bacterial communities, including a higher relative abundance of Firmicutes and a lower relative abundance of Proteobacteria, when compared to non-users. Non-users had a higher relative abundance of Actinomyces, Granulicatella, Haemophilus, Neisseria, Oribacterium, Prevotella, Pseudomonas, Rothia, and Veillonella in buccal swab samples, compared to tobacco users. While the most abundant bacterial genera were relatively constant over time, some species demonstrated significant shifts in relative abundance between the first and last time points. In addition, some opportunistic pathogens were detected among tobacco users including Neisseria subflava, Bulleidia moorei and Porphyromonas endodontalis. Overall, our results provide a more holistic understanding of the structure of oral bacterial communities in tobacco users compared to non-users.

ABSTRACT Lyme disease, caused by spirochetes in the Borrelia burgdorferi sensu lato clade within the Borrelia genus, is transmitted by Ixodes ticks and is currently the most prevalent and rapidly expanding tick-borne disease in Europe and North America. We report complete genome sequences of 47 isolates that encompass all established species in this clade while highlighting the diversity of the widespread human pathogenic species B. burgdorferi. A similar set of plasmids has been maintained throughout Borrelia divergence, indicating that they are a key adaptive feature of this genus. Phylogenetic reconstruction of all sequenced Borrelia genomes revealed the original divergence of Eurasian and North American lineages and subsequent dispersals that introduced B. garinii, B. bavariensis, B. lusitaniae, B. valaisiana, and B. afzelii from East Asia to Europe and B. burgdorferi and B. finlandensis from North America to Europe. Molecular phylogenies of the universally present core replicons (chromosome and cp26 and lp54 plasmids) are highly consistent, revealing a strong clonal structure. Nonetheless, numerous inconsistencies between the genome and gene phylogenies indicate species dispersal, genetic exchanges, and rapid sequence evolution at plasmid-borne loci, including key host-interacting lipoprotein genes. While localized recombination occurs uniformly on the main chromosome at a rate comparable to mutation, lipoprotein-encoding loci are recombination hotspots on the plasmids, suggesting adaptive maintenance of recombinant alleles at loci directly interacting with the host. We conclude that within- and between-species recombination facilitates adaptive sequence evolution of host-interacting lipoprotein loci and contributes to human virulence despite a genome-wide clonal structure of its natural populations.IMPORTANCELyme disease (also called Lyme borreliosis in Europe), a condition caused by spirochete bacteria of the genus Borrelia, transmitted by hard-bodied Ixodes ticks, is currently the most prevalent and rapidly expanding tick-borne disease in the United States and Europe. Borrelia interspecies and intraspecies genome comparisons of Lyme disease-related bacteria are essential to reconstruct their evolutionary origins, track epidemiological spread, identify molecular mechanisms of human pathogenicity, and design molecular and ecological approaches to disease prevention, diagnosis, and treatment. These Lyme disease-associated bacteria harbor complex genomes that encode many genes that do not have homologs in other organisms and are distributed across multiple linear and circular plasmids. The functional significance of most of the plasmid-borne genes and the multipartite genome organization itself remains unknown. Here we sequenced, assembled, and analyzed whole genomes of 47 Borrelia isolates from around the world, including multiple isolates of the human pathogenic species. Our analysis elucidates the evolutionary origins, historical migration, and sources of genomic variability of these clinically important pathogens. We have developed web-based software tools (BorreliaBase.org) to facilitate dissemination and continued comparative analysis of Borrelia genomes to identify determinants of human pathogenicity.

Abstract Background Chemotherapy is a primary treatment for cancer, but its efficacy is often limited by cancer-associated bacteria (CAB) that impair tumor suppressor functions. Our previous research found that Mycoplasma fermentans DnaK, a chaperone protein, impairs p53 activities, which are essential for most anti-cancer chemotherapeutic responses. Methods To investigate the role of DnaK in chemotherapy, we treated cancer cell lines with M. fermentans DnaK and then with commonly used p53-dependent anti-cancer drugs (cisplatin and 5FU). We evaluated the cells’ survival in the presence or absence of a DnaK-binding peptide (ARV-1502). We also validated our findings using primary tumor cells from a novel DnaK knock-in mouse model. To provide a broader context for the clinical significance of these findings, we investigated human primary cancer sequencing datasets from The Cancer Genome Atlas (TCGA). We identified F. nucleatum as a CAB carrying DnaK with an amino acid composition highly similar to M. fermentans DnaK. Therefore, we investigated the effect of F. nucleatum DnaK on the anti-cancer activity of cisplatin and 5FU. Results Our results show that both M. fermentans and F. nucleatum DnaKs reduce the effectiveness of cisplatin and 5FU. However, the use of ARV-1502 effectively restored the drugs' anti-cancer efficacy. Conclusions Our findings offer a practical framework for designing and implementing novel personalized anti-cancer strategies by targeting specific bacterial DnaKs in patients with poor response to chemotherapy, underscoring the potential for microbiome-based personalized cancer therapies.

Abstract Intrinsic metabolism shapes the immune environment associated with immune suppression and tolerance in settings such as organ transplantation and cancer. However, little is known about the metabolic activities in an immunosuppressive environment. In this study, we employed metagenomic, metabolomic, and immunological approaches to profile the early effects of the immunosuppressant drug tacrolimus, antibiotics, or both in gut lumen and circulation using a murine model. Tacrolimus induced rapid and profound alterations in metabolic activities within two days of treatment, prior to alterations in gut microbiota composition and structure. The metabolic profile and gut microbiome after seven days of treatment was distinct from that after two days of treatment, indicating continuous drug effects on both gut microbial ecosystem and host metabolism. The most affected taxonomic groups are Clostriales and Verrucomicrobiae (i.e., Akkermansia muciniphila), and the most affected metabolic pathways included a group of interconnected amino acids, bile acid conjugation, glucose homeostasis, and energy production. Highly correlated metabolic changes were observed between lumen and serum metabolism, supporting their significant interactions. Despite a small sample size, this study explored the largely uncharacterized microbial and metabolic events in an immunosuppressed environment and demonstrated that early changes in metabolic activities can have significant implications that may serve as antecedent biomarkers of immune activation or quiescence. To understand the intricate relationships among gut microbiome, metabolic activities, and immune cells in an immune suppressed environment is a prerequisite for developing strategies to monitor and optimize alloimmune responses that determine transplant outcomes.

Abstract Background The genomes of indigenous African cattle are composed of components with Middle Eastern (taurine) and South Asian (indicine) origins, providing a valuable model to study hybridization and to identify genetic barriers to gene flow. In this study, we analysed indigenous African cattle breeds as models of hybrid zones, considering taurine and indicine samples as ancestors. In a genomic cline analysis of whole-genome sequence data, we considered over 8 million variants from 144 animals, which allows for fine-mapping of potential genomic incompatibilities at high resolution across the genome. Results We identified several thousand variants that had significantly steep clines (‘SCV’) across the whole genome, indicating restricted introgression. Some of the SCV were clustered into extended regions, with the longest on chromosome 7, spanning 725 kb and including 27 genes. We found that variants with a high phenotypic impact (e.g. indels, intra-genic and missense variants) likely represent greater genetic barriers to gene flow. Furthermore, our findings provide evidence that a large proportion of breed differentiation in African cattle could be linked to genomic incompatibilities and reproductive isolation. Functional evaluation of genes with SCV suggest that mitonuclear incompatibilities and genes associated with fitness (e.g. resistance to paratuberculosis) could account for restricted gene flow in indigenous African cattle. Conclusions To our knowledge, this is the first time genomic cline analysis has been applied to identify restricted introgression in the genomes of indigenous African cattle and the results provide extended insights into mechanisms (e.g. genomic incompatibilities) contributing to hybrid differentiation. These results have important implications for our understanding of genetic incompatibilities and reproductive isolation and provide important insights into the impact of cross-breeding cattle with the aim of producing offspring that are both hardy and productive.

A conventional hydrocyclones is a versatile equipment with a high processing capacity and low maintenance cost. Currently, several studies aim to alter the typical structure of the conventional hydrocyclone in order to modify its performance and purpose. For this, filtering hydrocyclones have emerged, where a porous membrane replaces the conic or cylindrical wall. During the operation of this equipment, in addition to the traditionally observed streams (feed, underflow, and overflow), there is a liquid stream resulting from the filtration process, commonly referred to as filtrate. This work proposes to numerically investigate the solid particle/liquid water separation process in a filtering hydrocyclone using the commercial software Ansys CFX® 15.0. The proposed mathematical model for the study considers three-dimensional, steady state and turbulent flow, using the Eulerian–Eulerian approach and the Shear Stress Transport (SST) turbulence model. This study presents and analyzes the volume fraction, velocity, and pressure fields, along with flowlines and velocity profiles. The results indicate that the proposed model effectively captures the fluid dynamic behavior within the filtering hydrocyclone, highlighting higher pressures near the porous membrane and a higher concentration of solid particles in the conical region, with water being more concentrated in the cylindrical part of the hydrocyclone. Additionally, the findings show that the volumetric flow rate of the filtrate significantly influences the internal flow dynamics, with conventional hydrocyclones demonstrating higher pressure gradients compared to the proposed filtering hydrocyclone.

Abstract Advanced treated municipal wastewater is an important alternative water source for agricultural irrigation. However, the possible persistence of chemical and microbiological contaminants in these waters raise potential safety concerns with regard to reusing treated wastewater for food crop irrigation. Two low-cost and environmentally-friendly filter media, biochar (BC) and zero-valent iron (ZVI), have attracted great interest in terms of treating reused water. Here, we evaluated the efficacy of BC-, nanosilver-amended biochar- (Ag-BC) and ZVI-sand filters, in reducing contaminants of emerging concern (CECs), Escherichia coli (E. coli) and total bacterial diversity from wastewater effluent. Six experiments were conducted with control quartz sand and sand columns containing BC, Ag-BC, ZVI, BC with ZVI, or Ag-BC with ZVI. After filtration, Ag-BC, ZVI, BC with ZVI and Ag-BC with ZVI demonstrated more than 90% (> 1 log) removal of E. coli from wastewater samples, while BC, Ag-BC, BC with ZVI and Ag-BC with ZVI also demonstrated efficient removal of tested CECs. Lower bacterial diversity was also observed after filtration; however, differences were marginally significant. In addition, significantly (p

Background: Pregnant women's understanding of maternal health information is critical in the application of information for pregnancy care, birth preparedness, and newborn care. However, the influence of socio-demographic characteristics on pregnant women's understanding of maternal health information is least understood. Purpose: The study examined the influence of social and demographic characteristics on pregnant women's comprehension of maternal health information. Methodology: A mixed research approach and descriptive cross-sectional design were applied. Questionnaires and focus group discussions were used to collect data from 132 pregnant women and 8 nurses/midwives. Data were analyzed using IBM SPSS version 21 and thematic analysis. Findings: Most of the pregnant women had a good ability to comprehend maternal health information. Level of education was found to influence the reading ability and language understanding ability of the pregnant women unlike age, marital status, income, and occupation. Age, level of education, monthly income, occupation, and marital status were found to influence the ability of pregnant women to understand maternal health information. Conclusion: The socio-demographic characteristics of pregnant women have an influence on their ability to comprehend maternal health information.

ABSTRACTBifidobacterium is a widely distributed commensal bacterial genus that displays beneficial pro-homeostatic and anti-inflammatory immunomodulatory properties. Depletion or absence of Bifidobacterium in humans and model organisms is associated with autoimmune responses and impaired immune homeostasis. At the cellular level, Bifidobacterium upregulates suppressive regulatory T cells, maintains intestinal barrier function, modulates dendritic cell and macrophage activity, and dampens intestinal Th2 and Th17 programs. While there has been a large volume of literature characterizing the probiotic properties of various Bifidobacterial species, the likely multifactorial mechanisms underlying these effects remain elusive, in particular, its immune tolerogenic effect. However, recent work has shed light on Bifidobacterium surface structural polysaccharide and protein elements, as well as its metabolic products, as commensal mediators of immune homeostasis. This review aims to discuss several mechanisms Bifidobacterium utilizes for immune modulation as well as their indirect impact on the regulation of gut microbiome structure and function, from structural molecules to produced metabolites. These mechanisms are pertinent to an increasingly networked understanding of immune tolerance and homeostasis in health and disease.

Lyme disease is a multisystem disorder primarily caused by Borrelia burgdorferi sensu lato. However, B. garinii, which has been identified on islands off the coast of Newfoundland and Labrador, Canada, is a cause of Lyme disease in Eurasia. We report isolation and whole-genome nucleotide sequencing of a B. garinii isolate from a cotton mouse (Peromyscus gossypinus) in South Carolina, USA. We identified a second B. garinii isolate from the same repository. Phylogenetic analysis does not associate these isolates with the previously described isolates of B. garinii from Canada.

Abstract The beneficial effects attributed to Bifidobacterium are largely attributed to their immunomodulatory capabilities, which are likely to be species- and even strain-specific. However, their strain-specificity in direct and indirect immune modulation remain largely uncharacterized. We have shown that B. pseudolongum UMB-MBP-01, a murine isolate strain, is capable of suppressing inflammation and reducing fibrosis in vivo. To ascertain the mechanism driving this activity and to determine if it is specific to UMB-MBP-01, we compared it to a porcine tropic strain B. pseudolongum ATCC25526 using a combination of cell culture and in vivo experimentation and comparative genomics approaches. Despite many shared features, we demonstrate that these two strains possess distinct genetic repertoires in carbohydrate assimilation, differential activation signatures and cytokine responses signatures in innate immune cells, and differential effects on lymph node morphology with unique local and systemic leukocyte distribution. Importantly, the administration of each B. pseudolongum strain resulted in major divergence in the structure, composition, and function of gut microbiota. This was accompanied by markedly different changes in intestinal transcriptional activities, suggesting strain-specific modulation of the endogenous gut microbiota as a key to immune modulatory host responses. Our study demonstrated a single probiotic strain can influence local, regional, and systemic immunity through both innate and adaptive pathways in a strain-specific manner. It highlights the importance to investigate both the endogenous gut microbiome and the intestinal responses in response to probiotic supplementation, which underpins the mechanisms through which the probiotic strains drive the strain-specific effect to impact health outcomes.

The use of synthetic compounds as corrosion inhibitor is diminishing due to strict environmental regulations and toxic effects of the compounds on human beings and the environment. Consequently, there exists the need to develop a new class of corrosion inhibitors with low toxicity, eco-friendliness and good efficiency. Therefore, this study investigates the inhibitory potential of Acalypha wilkesiana (AWAE) extract on mild steel in 1M HCl using weight loss method. Experiments were performed by varying immersion period, concentration of the inhibitor and temperature. The results obtained showed a decrease in the corrosion rate of mild steel as the extract concentration increases while the inhibition efficiency increases with extract concentration. Effect of temperature on the inhibition efficiency showed an increase in inhibition efficiency with rise in temperature. The adsorption of the inhibitor on the mild steel surface in acid solution was found to obey the Langmuir adsorption isotherm. Calculated thermodynamic parameters revealed that the mechanism of the corrosion inhibition may be due to the chemical adsorption of the phyto-constituents on the surface of mild steel. Kinetic treatment of the data followed a first order reaction and the half-life values of the metal increase with increase in the extract concentration. Preliminary investigation of the phytochemical constituents showed that Acalypha wilkesiana contains tannin, flavonoid, saponin, alkaloid, steroid , terpenoid and some other compounds in trace quantity. FTIR results indicate that AWAE contains O and N atoms in functional groups and aromatic ring which meet the general consideration of a typical corrosion inhibitor.

BackgroundThe United States' (U.S.) initiative to End the HIV Epidemic aims to reduce new HIV infections in areas of high HIV prevalence. Despite national efforts to reduce HIV incidence, cisgender women continue to represent approximately one out of every five new HIV diagnoses in the U.S. Taking pre-exposure prophylaxis (PrEP) is an effective HIV prevention strategy; however, PrEP initiation among cisgender women is suboptimal, with only 10% of eligible women receiving PrEP prescriptions in 2019.MethodsWe designed a trial to test the effectiveness of interventions to increase PrEP initiation, while evaluating the implementation strategy (hybrid type II trial) in seven obstetrics and gynecology (OB/GYN) clinics (two federally qualified health centers, three community-based, and two academic) in Baltimore, Maryland. A total of 42 OB/GYN providers will be enrolled and randomized (1:1:1) into one of three clinical trial arms (standard of care, patient-level intervention, or multi-level intervention). Eligible patients of enrolled providers will receive a sexual health questionnaire before their appointment through the electronic health record’s (EHR) patient portal. The questionnaire will be scored in three tiers (low, moderate, and high) to assess HIV risk. Patients at low risk will be offered an HIV test only, while those who score medium or high risk will be included in the clinical trial and assigned to the clinical trial arm associated with their provider. Differences in PrEP initiation, our primary outcome, across the three arms will be analyzed using generalized linear mixed-effect models with logistic regression. We will adjust results for demographic differences observed between arms and examine PrEP initiation stratified by patient’s and provider’s race and ethnicity.Additionally, a comprehensive economic analysis for each intervention will be conducted.DiscussionWe hypothesize that gathering information on sensitive sexual behaviors electronically, communicating HIV risk in an understandable and relatable format to patients and OB/GYN providers, and deploying EHR alerts will increase PrEP initiation and HIV testing.Trial RegistrationThe trial is registered with ClinicalTrials.gov (NCT05412433) on 09 June 2022. https://clinicaltrials.gov/ct2/show/NCT05412433?term=NCT05412433&draw=2&rank=1.

In the terrestrial carbon cycle is very relevant to identify the influence of soil in the CO2 released to the atmosphere, which is linked to multiple biotic and abiotic drivers. Arid ecosystems dominate the trend and interannual variability of the land CO2 sink. This pattern is mainly controlled by temperature, precipitation, and shortwave radiation. Thus, these environments are characterized by a wide variability of water availability, which causes the CO2 efflux to be highly variable in time, challenging our model capacities. This study aims to understand the ecosystem CO2 fluxes and their controlling mechanisms from the Chihuahuan Desert in Northeast Mexico. We explore the average contribution of the Rsoil (1.30 mmol m-2 s‑1) to Reco (1.76 mmol m-2 s‑1), while identifying the controlling mechanisms of both on an annual scale. The structural equation model constructed showed a good fit for the data, explaining 50% and 93% of the annual variance of Rsoil and Reco, respectively. According to this model, Rsoil was mainly controlled by the air temperature, and Reco by soil water content. Unexpectedly, vapor pressure deficit was the most weight variable with a direct negative effect on Reco, supporting the idea that the vegetation component has a crucial role in the CO2 efflux of this ecosystem. This study highlights the importance of include multiple factors in the models of the C cycle.

Arid and semi-arid ecosystems dominate the RS variability due to the multiple changing factors that control it. Consequently, any variation, in addition to climate change and land use change, impacts the concentration of CO2 in the atmosphere. Here, the effect of the interannual variation and the controlling factors of RS in native xeric shrublands and agricultural systems is investigated. This study was conducted in four sites per condition for two years (2019 to 2020), where RS and the soil properties were measured. The RS presented a higher variation in the xeric shrubland. The agricultural plots showed the highest RS (0.33 g CO2 m−2 hr−1) compared to the xeric shrubland (0.12 g CO2 m−2 hr−1). The soil water content was the main controlling variable for RS in both land uses. However, soil temperature affected RS only in agricultural plots. The variation in the RS under different land uses confirms that changes in the soil and environmental conditions (i.e., season) control the RS. In addition, if current management practices are maintained in agricultural sites and under a temperature increase scenario, a significant increase in the RS rate is expected.

Context and background Land use plans have been considered as a solution to land use problems and hence enhance ecological, economic and social sustainability of land use. Appropriateness of land use plans and hence its potential for adherence may rely on sufficiency of zones allocated for different land uses. Goal and Objectives: This study was designed to empirically identify land use implementation problems and suggest solutions relevant to the land users, the government, planners and other stakeholders. Specifically, the study assesses: (1) The extent to which the land use zones cover all zones needed by the stakeholders and; (2) Reasons for levels of sufficiency of the allocated land use zones. Methodology: Data were collected through household survey of 120 respondents from two villages, key informants, focus group discussions and field observation survey while secondary data were collected through review of guidelines for land use planning, village land use plans, district land use framework, books and journals. Information used to assess sufficiency of land use zones used in Village Land Use Plans (VLUP) from household survey and village records were descriptively analyzed. Results: The implementation of village land use plans was not done as expected. Land use zones were insufficient in terms of the allocated size and needs within the zones for current and future situation because of increasing population, overstocking, and lack of infrastructure necessary within specific zones. Other factors included inadequate consideration for uncertainties in population projection standard, unclear zoning regulation and discrepancy in population data. Based on the findings and conclusions, this study makes the following recommendations. First, the National Land Use Planning Commission should devise mechanisms to ensure that all the six steps of land use planning are completed towards implementable land use plans. Secondly, the national land use planning commission should review zoning standards to sufficiently allocate the land use zones. The population projections used for future allocation of land had influence on the sufficiency of the zones where the rate of population increase is assumed to be fixed throughout the ten years implementation period without consideration of uncertainties. It is worth incorporating GIS to establish trend of land use and forecast future land use to sufficiently allocate land during the 10 years lifespan of the VLUP. Thirdly, the national land use planning commission need to validate spatial data and population data at village level to avoid discrepancies which affect implementation of the village land use plans.

Abstract Nontraditional irrigation water sources (e.g., recycled water, brackish water) may harbor human pathogens, including Vibrio spp., that could be present in a viable-but-nonculturable (VBNC) state, stymieing current culture-based detection methods. To overcome this challenge, we coupled 5-bromo-2′-deoxyuridine (BrdU) labeling, enrichment techniques, and 16S rRNA sequencing to identify metabolically-active Vibrio spp. in nontraditional irrigation water (recycled water, pond water, non-tidal freshwater, and tidal brackish water). Our coupled BrdU-labeling and sequencing approach revealed the presence of metabolically-active Vibrio spp. at all sampling sites. Whereas, the culture-based method only detected vibrios at three of the four sites. We observed the presence of V. cholerae, V. vulnificus, and V. parahaemolyticus using both methods, while V. aesturianus and V. shilonii were detected only through our labeling/sequencing approach. Multiple other pathogens of concern to human health were also identified through our labeling/sequencing approach including P. shigelloides, B. cereus and E. cloacae. Most importantly, 16S rRNA sequencing of BrdU-labeled samples resulted in Vibrio spp. detection even when our culture-based methods resulted in negative detection. This suggests that our novel approach can effectively detect metabolically-active Vibrio spp. that may have been present in a VBNC state, refining our understanding of the prevalence of vibrios in nontraditional irrigation waters.

Abstract Background Removals caused by both natural and anthropogenic drivers such as logging and fire in miombo woodlands causes substantial carbon emissions. Here we present drivers and their effects on the variations on the number of stems and aboveground carbon (AGC) removals based on an analysis of Tanzania’s national forest inventory (NFI) data extracted from the National Forest Resources Assessment and Monitoring (NAFORMA) database using allometric models that utilize stump diameter as the sole predictor. Results Drivers of AGC removals in miombo woodlands of mainland Tanzania in order of importance were timber, fire, shifting cultivation, charcoal, natural death, firewood collection, poles, grazing by wildlife animals, carvings, grazing by domestic animals, and mining. The average number of stems and AGC removals by driver ranged from 0.006 to 16.587 stems ha−1 year−1 and 0.0–1.273 tCha−1 year−1 respectively. Furthermore, charcoal, shifting cultivation and fuelwood caused higher tree removals as opposed to timber, natural death and fire that accounted for higher AGC removals. Conclusions Drivers caused substantial effects on the number of stems and carbon removals. Increased mitigation efforts in addressing removals by timber, fires, shifting cultivation, charcoal and natural death would be effective in mitigating degradation in miombo woodlands of Tanzania. Additionally, site-specific studies need to be conducted to bring information that would be used for managing woodlands at local levels. This kind of study need to be conducted in other vegetation types like montane and Mangrove forest at national scale in Tanzania.

The service learning in multimedia skills for teenagers and young people as training program for the community was conducted through the expertise of Bachelor Program in Computer Engineering, Faculty of Engineering, Universitas Kristen Maranatha, for Gereja Kristen Pasundan (GKP) Cideres. The purpose of this training was to equip people with skills or knowledge that can be useful in daily activities. As this training was given during the Covid-19 pandemic, by requirement of health and safety protocols, the service learning was implemented online via the internet in a form of webinar. Online digital activities are increasingly important as conventional physical interactions and communication in work or study environment become unsafe and to be avoided or to be done less often than usual. Unfortunately for most people, technical skills for working or studying online are not yet adequate, especially in multimedia using some recent tools and softwares. So in this program, basic multimedia producing skills using Microsoft Photos (video editing feature) and Microsoft Power Point were introduced. The end result of the training was satisfactory, as those who participate are being able to produce simple digital multimedia presentation using the aforementioned tools and softwares. The training program was held online on November 28th, 2020 via Zoom Meeting Application, and was made possible thanks to the collaboration of Bachelor Program in Computer Engineering with the Directorate of Student and Alumni Affairs, Universitas Kristen Maranatha. Keywords: Covid-19 pandemic; multimedia; service learning

Abstract Background The New Guinean archipelago has been shaped by millions of years of plate tectonic activity combined with long-term fluctuations in climate and sea level. These processes combined with New Guinea’s location at the tectonic junction between the Australian and Pacific plates are inherently linked to the evolution of its rich endemic biota. With the advent of molecular phylogenetics and an increasing amount of geological data, the field of New Guinean biogeography begins to be reinvigorated. Results We inferred a comprehensive dated molecular phylogeny of endemic diving beetles to test historical hypotheses pertaining to the evolution of the New Guinean biota. We used geospatial analysis techniques to compare our phylogenetic results with a newly developed geological terrane map of New Guinea as well as the altitudinal and geographic range of species ( https://arcg.is/189zmz ). Our divergence time estimations indicate a crown age (early diversification) for New Guinea Exocelina beetles in the mid-Miocene ca. 17 Ma, when the New Guinean orogeny was at an early stage. Geographic and geological ancestral state reconstructions suggest an origin of Exocelina ancestors on the eastern part of the New Guinean central range on basement rocks (with a shared affinity with the Australian Plate). Our results do not support the hypothesis of ancestors migrating to the northern margin of the Australian Plate from Pacific terranes that incrementally accreted to New Guinea over time. However, our analyses support to some extent a scenario in which Exocelina ancestors would have been able to colonize back and forth between the amalgamated Australian and Pacific terranes from the Miocene onwards. Our reconstructions also do not support an origin on ultramafic or ophiolite rocks that have been colonized much later in the evolution of the radiation. Macroevolutionary analyses do not support the hypothesis of heterogeneous diversification rates throughout the evolution of this radiation, suggesting instead a continuous slowdown in speciation. Conclusions Overall, our geospatial analysis approach to investigate the links between the location and evolution of New Guinea’s biota with the underlying geology sheds a new light on the patterns and processes of lineage diversification in this exceedingly diverse region of the planet.

This present work revealed the isotherm, kinetic, and thermodynamic behaviour of methylene blue (MB) dye adsorbed onto acidic activated carbon (AAC) and base activated carbon (BAC) prepared from Raphia taedigera seed by carbonization and chemical activation. AAC and BAC were activated with sulphuric acid and sodium hydroxide respectively. Batch equilibrium studies were done under different experimental conditions such as MB dye concentration and temperature. The equilibrium data were modelled using Langmuir, Freundlich, Elovich, Temkin and Dubinin-Radushkevich isotherms. The Langmuir isotherm model best describes the uptake of MB dye onto AAC and BAC with R2 > 0.998 in all cases. The pseudo-first-order, pseudo-second-order and intra-particle diffusion equations were used to evaluate the kinetic properties. It was observed that the adsorption of MB dye onto the two activated carbons could best be described by the pseudo-second order equation with 0.999 < R2 ≤ 1. Thermodynamic parameters such as Gibbs free energy (ΔG0), standard enthalpy (ΔH0), standard entropy (ΔS0), and activation energy (Ea) were determined. The results of ΔG0indicated a spontaneous and feasible for AAC and non-spontaneous but feasible for BAC. Results of ΔH0 confirmed that the adsorption of MB onto AAC and BAC are endothermic and physical in nature. It can be concluded that AAC and BAC prepared from Raphia taedigera seed could be used as low-cost adsorbent for the removal of MB dye from the wastewater.

Abstract Background Ponds are important freshwater habitats that support both human and environmental activities. However, relative to their larger counterparts (e.g. rivers, lakes), ponds are understudied, especially with regard to their microbial communities. Our study aimed to fill this knowledge gap by using culture-independent, high-throughput sequencing to assess the dynamics, taxonomy, and functionality of bacterial and viral communities in a freshwater agricultural pond. Results Water samples (n = 14) were collected from a Mid-Atlantic agricultural pond between June 2017 and May 2018 and filtered sequentially through 1 and 0.2 μm filter membranes. Total DNA was then extracted from each filter, pooled, and subjected to 16S rRNA gene and shotgun sequencing on the Illumina HiSeq 2500 platform. Additionally, on eight occasions water filtrates were processed for viral metagenomes (viromes) using chemical concentration and then shotgun sequenced. A ubiquitous freshwater phylum, Proteobacteria was abundant at all sampling dates throughout the year. However, environmental characteristics appeared to drive the structure of the community. For instance, the abundance of Cyanobacteria (e.g. Nostoc) increased with rising water temperatures, while a storm event appeared to trigger an increase in overall bacterial diversity, as well as the relative abundance of Bacteroidetes. This event was also associated with an increase in the number of antibiotic resistance genes. The viral fractions were dominated by dsDNA of the order Caudovirales, namely Siphoviridae and Myovirdae. Conclusions Overall, this study provides one of the largest datasets on pond water microbial ecology to date, revealing seasonal trends in the microbial taxonomic composition and functional potential.

African women have a history of entrepreneurship dating back to the pre-colonial era, but in this 21st century, women in Africa are assessed by the World Bank to be at the lower deck of economic breakthrough in comparison to men. Although both men and women operate in the same business climate, women are usually poorer and are harder hit by adverse economic situations. Oftentimes, they self-develop their businesses and skills; they also seek, create and self-fund opportunities, but these efforts are usually met with huge obstacles and barriers. Therefore, this study aims to identify some of the obstacles and barriers plaguing the success and growth of women entrepreneurial activities and proffer solutions that could ameliorate the problems. The study uses a qualitative case study research design to uncover the experiences of women in a rural community of southwestern Nigeria. The data for analysis were collected through twenty in-depth interviews, two participants observations, and four focus group discussions. Some documents were analyzed to triangulate the data sources to ensure credibility. Findings include obstacles such as a traditional patriarchal culture that inhibits women from achieving personal development, complete subordination to male domination, colonial vestiges that gendered entrepreneurship in Africa, and lack of support by male-dominated government. The implication of these is that women continue to struggle to develop their entrepreneurial activities without much breakthrough. They remain economically impoverished and suffer greatly to care for themselves and their families, and to be part of social development. The study recommends that relevant organizations and government continue to work to develop strategies to remove these barriers.

PurposeDefine incidence and risk factors of osteonecrosis of the jaw (ONJ) and explore oral microbial signatures and host immune response as reflected by cytokine changes in saliva and serum in multiple myeloma (MM) patients on bisphosphate (BP) therapy.Patients and MethodsA single center observational prospective study of MM patients (n = 110) on >2 years of BP, none had ONJ at enrollment. Patients were followed every 3 months for 18 months with clinical/dental examination and serial measurements of inflammatory cytokines, bone turnover markers, and angiogenic growth factors. Oral microbiota was characterized by sequencing of 16S rRNA gene from saliva.ResultsOver the study period 14 patients (13%) developed BRONJ, at a median of 5.7 years (95% CI: 1.9–12.0) from MM diagnosis. Chronic periodontal disease was the main clinically observed risk factor. Oral microbial profiling revealed lower bacterial richness/diversity in BRONJ. Streptococcus intermedius, S. mutans, and S. perioris were abundant in controls; S. sonstellatus and S anginosus were prevalent in BRONJ. In the saliva, at baseline patients who developed BRONJ had higher levels of MIP-1β; TNF-α and IL-6 compared to those without BRONJ, cytokine profile consistent with M-1 macrophage activation. In the serum, patients with BRONJ have significantly lower levels of TGF beta and VEGF over the study period.ConclusionPeriodontal disease associated with low microbial diversity and predominance of invasive species with a proinflammatory cytokine profile leading to tissue damage and alteration of immunity seems to be the main culprit in pathogenesis of BRONJ.

ObjectiveTo characterize the microbial communities of the anterior nares (nose) and posterior pharynx (throat) of adults dwelling in the community and in nursing homes before and after treatment with intranasal mupirocin.MethodsStaphylococcus aureus-colonized adults were recruited from the community (n = 25) and from nursing homes (n = 7). S. aureus colonization was confirmed using cultures. Participants had specimens taken from nose and throat for S. aureus quantitation using quantitative PCR for the nuc gene and bacterial profiling using 16S rRNA gene sequencing over 12 weeks. After two baseline study visits 4 weeks apart, participants received intranasal mupirocin for 5 days with 3 further visits over a 8 week follow-up period.ResultsWe found a decrease in the absolute abundance of S. aureus in the nose for 8 weeks after mupirocin (1693 vs 141 fg/ul, p = 0.047). Mupirocin caused a statistically significant disruption in bacterial communities of the nose and throat after 1 week, which was no longer detected after 8 weeks. Bacterial community profiling demonstrated that there was a decrease in the relative abundance of S. aureus (8% vs 0.3%, pConclusionsDecolonization with mupirocin leads to a sustained effect on absolute and relative abundance of S. aureus but not for other bacteria in the nose. This demonstrates that a short course of mupirocin selectively decreases S. aureus in the nose for up to 8 weeks.

The etiology of vulvar lichen sclerosus (LS) remains unclear; however, alterations in cutaneous and gut microbiota may be contributing to the pathogenesis of this inflammatory condition. To explore this hypothesis, we conducted a pilot case-control study, obtaining dermal swab and stool samples from prepubertal girls with vulvar LS (n = 5), girls with nonspecific vulvovaginitis (n = 5), and healthy controls (n = 3). Samples (n = 56) were subjected to total DNA extractions. Resulting DNA was purified, subjected to PCR (targeting the V3V4 region of the 16S rRNA gene), sequenced, and analyzed using QIIME, MetagenomeSeq, and DESeq2 software packages. Our findings showed that there were significant differences in the cutaneous and gut microbiotas of girls with LS compared to controls. On the skin, girls with LS had a statistically significantly higher relative abundance of Porphyromonas spp., Parvimonas spp., Peptoniphilus spp., Prevotella spp., Dialister spp., and Peptostreptococcus spp., but a lower relative abundance of Cornyebacterium compared to the control group. In the gut samples, girls with LS had a significantly higher relative abundance of Dialister spp., Clostridiales spp., Paraprevotella spp., Escherichia coli, Bifidobacterium adolescentis, and Akkermansia muciniphila, and a lower relative abundance of Roseburia faecis and Ruminococcus bromii compared to controls. These results suggest a potential association between cutaneous and gut dysbiosis and pediatric vulvar LS. Future studies involving larger samples sizes are warranted to further evaluate this association.

The increasing use of reclaimed water for irrigation in areas lacking access to advanced wastewater treatment and reclaimed water distribution systems calls for an examination of irrigation-site-based treatment technologies that can improve the quality of this alternative water source. To address this need, we investigated the impact of zero-valent iron (ZVI)-sand filtration on the bacterial community structure and functional potential of conventionally treated reclaimed water utilized in downstream applications. Over a 2-month period, reclaimed water was collected from a tertiary wastewater treatment plant in the Mid-Atlantic, U.S. and trucked to our greenhouse facility. The water was stored in rain barrels and then filtered through one ZVI-sand filter every 5 days. Filtrate was then subjected to enumeration, phylotyping, shiga toxin screening, and antimicrobial susceptibility testing of Escherichia coli. Aliquots of filtrate were also DNA extracted, and purified DNA was subjected to 16S rRNA gene sequencing and metagenomic shotgun sequencing. The genera Dechloromonas, Desulfotomaculum, Leptonema, and Thermomonas, which contain denitrifying and sulfate reducing species, commonly used in bioremediation, and known to increase the inherent reactivity of ZVI, were significantly more relatively abundant in ZVI-sand filtered reclaimed water compared to reclaimed water. The concentration of E. coli in ZVI-sand filtered reclaimed water was significantly lower compared to that of reclaimed water, and cefoxitin- and tetracycline-resistant E. coli were undetectable after ZVI-sand filtration. ZVI-sand filtration reduced the occurrence of human as well as plant pathogenic genera (Aeromonas, Mycobacterium, Shewanella, Acidovorax, Agrobacterium, and Clavibacter) but increased the proportion of Azospira, a nitrogen fixing bacterial genera, in the microbial community. Our exploratory functional analysis showed a modest non-significant increase in the proportion of open reading frames for genes associated with iron uptake, oxidative stress, as well as defense and repair mechanisms after ZVI-sand filtration. These data indicate an iron rich environment in the filter causing an oxidative stress response by the bacterial community present in the reclaimed water. Our findings demonstrate that ZVI-sand filtration effectively filters conventionally treated reclaimed water. Longer-term, field-based studies are needed to evaluate the effectiveness of the filter in agricultural settings and inform the development of future agricultural water reuse regulations.

The expansion of grassland plant diversity is thought to have facilitated diversification of herbivorous insects. Here, the authors show opposing evolutionary dynamics in a clade of African grasses and associated stemborers, opposing the hypothesis about grasslands as a 'cradle' of herbivore diversity.

Abstract Background Chronic rhinosinusitis is an inflammatory disorder in which the role of bacteria remains uncertain. While sinus outflow obstruction is often an initiating event, mucosal inflammation and dysbiosis may persist or develop in sinuses with widely patent surgical openings. Understanding of the relationship between dysbiosis and chronic sinus inflammation is obfuscated by inter-individual microbiota variability and likely intra-individual temporal variation that has yet to be defined. In this study, long-term microbiota stability is investigated within surgically-opened maxillary sinuses of individuals with and without sinus inflammatory disease. Methods Maxillary sinus swabs were performed in 35 subjects with longstanding maxillary antrostomies. Subjects with and without active chronic maxillary sinusitis were included. Repeat swabs were obtained from the same sinuses after a prolonged interval (mean 719 ± 383 days). Patients were categorized based on the inflammatory status of the sinus mucosa at times of sample collection, as assessed by nasal endoscopy. Total DNA from swab eluents was extracted, and the microbiota characterized using 16S rRNA gene sequencing followed by taxonomic classification. Prevalence and abundance of genera were determined by analysis of 16S rRNA gene sequences. Taxa were identified that were stably present between two time points in individual subjects. Results The overall proportion of stable taxa across time points was 24.5 ± 10.6%. This stability index was consistent across patient groups and not correlated with clinical parameters. Highly prevalent taxa, including Staphylococcus, Corynebacterium, Propionibacterium, and Pseudomonas, were often stably present, but varied in relative abundance. Janthinobacterium, Enterobacter, Lactobacillus, and Acinetobacter were prevalent and moderately abundant taxa in healthy sinuses, but not in inflamed sinuses. Moraxella and Haemophilus were present at low prevalence and proportional abundance in chronically or intermittently inflamed sinuses, but not in healthy sinuses. Conclusions A relatively small component of the post-antrostomy maxillary sinus microbiota exhibits long-term stability in individual subjects. Stable bacteria include a limited number of highly prevalent and a larger number of lower prevalence taxa, which vary widely in proportional abundance. The concept of individual-specific core sinus microbiota, durable over time and medical therapy, but fluctuating in proportional abundance, has implications for understanding the role of bacteria in CRS pathogenesis.

Abstract Background Butterflies (Papilionoidea) are perhaps the most charismatic insect lineage, yet phylogenetic relationships among them remain incompletely studied and controversial. This is especially true for skippers (Hesperiidae), one of the most species-rich and poorly studied butterfly families. Methods To infer a robust phylogenomic hypothesis for Hesperiidae, we sequenced nearly 400 loci using Anchored Hybrid Enrichment and sampled all tribes and more than 120 genera of skippers. Molecular datasets were analyzed using maximum-likelihood, parsimony and coalescent multi-species phylogenetic methods. Results All analyses converged on a novel, robust phylogenetic hypothesis for skippers. Different optimality criteria and methodologies recovered almost identical phylogenetic trees with strong nodal support at nearly all nodes and all taxonomic levels. Our results support Coeliadinae as the sister group to the remaining skippers, the monotypic Euschemoninae as the sister group to all other subfamilies but Coeliadinae, and the monophyly of Eudaminae plus Pyrginae. Within Pyrginae, Celaenorrhinini and Tagiadini are sister groups, the Neotropical firetips, Pyrrhopygini, are sister to all other tribes but Celaenorrhinini and Tagiadini. Achlyodini is recovered as the sister group to Carcharodini, and Erynnini as sister group to Pyrgini. Within the grass skippers (Hesperiinae), there is strong support for the monophyly of Aeromachini plus remaining Hesperiinae. The giant skippers (Agathymus and Megathymus) once classified as a subfamily, are recovered as monophyletic with strong support, but are deeply nested within Hesperiinae. Conclusions Anchored Hybrid Enrichment sequencing resulted in a large amount of data that built the foundation for a new, robust evolutionary tree of skippers. The newly inferred phylogenetic tree resolves long-standing systematic issues and changes our understanding of the skipper tree of life. These resultsenhance understanding of the evolution of one of the most species-rich butterfly families.