The Anopheles coluzzii range extends into Kenya: detection, insecticide resistance profiles and population genetic structure in relation to conspecific populations in West and Central Africa.

BACKGROUND: Anopheles coluzzii is a primary vector of malaria found in West and Central Africa, but its presence has hitherto never been documented in Kenya. A thorough understanding of vector bionomics is important as it enables the implementation of targeted and effective vector control interventions. Malaria vector surveillance efforts in the country have tended to focus on historically known primary vectors. The current study sought to determine the taxonomic status of samples collected from five different malaria epidemiological zones in Kenya as well as describe the population genetic structure and insecticide resistance profiles in relation to other An. coluzzii populations. METHODS: Mosquitoes were sampled as larvae from Busia, Kwale, Turkana, Kirinyaga and Kiambu counties, representing the range of malaria endemicities in Kenya, in 2019 and 2021 and emergent adults analysed using Whole Genome Sequencing (WGS) data processed in accordance with the Anopheles gambiae 1000 Genomes Project phase 3. Where available, historical samples from the same sites were included for WGS. Comparisons were made with An. coluzzii cohorts from West and Central Africa. RESULTS: This study reports the detection of An. coluzzii for the first time in Kenya. The species was detected in Turkana County across all three time points from which samples were analyzed and its presence confirmed through taxonomic analysis. Additionally, there was a lack of strong population genetic differentiation between An. coluzzii from Kenya and those from the more northerly regions of West and Central Africa, suggesting they represent a connected extension to the known species range. Mutations associated with target-site resistance to DDT and pyrethroids and metabolic resistance to DDT were found at high frequencies up to 64%. The profile and frequencies of the variants observed were similar to An. coluzzii from West and Central Africa but the ace-1 mutation linked to organophosphate and carbamate resistance present in An. coluzzii from coastal West Africa was absent in Kenya. CONCLUSIONS: These findings emphasize the need for the incorporation of genomics in comprehensive and routine vector surveillance to inform on the range of malaria vector species, and their insecticide resistance status to inform the choice of effective vector control approaches.

Repellent Effects of Selected Organic Leaf Extracts of Tithonia diversifolia (Hemsl.) A. Gray and Vernonia lasiopus (O. Hoffman) against Sitophilus zeamais Motschulsky (Coleoptera: Curculionidae).

INTRODUCTION: Sitophilus zeamais infestation is among the major setbacks to sustainable maize farming and availability. It causes an estimated annual loss of 5-10% and 20-30% of the total maize grains loss in the temperate and tropical zones, respectively. Although synthetic pesticides are quick and effective in managing crop pests, their overuse and misuse is discouraged due to their detrimental effects on human and environment. Natural pesticidal products that are extracted from plants are particularly gaining importance as an alternative to synthetic pesticides. They are available, easily biodegraded and have low toxicity to nontarget organisms. Most botanical pesticides act on insects by repelling them away from the crops in the field or in the stores. Therefore, this study aimed to determine repellency potential of organic leaf extracts of Tithonia diversifolia and Vernonia lasiopus on S. zeamais. Materials and methods. The phytochemical profile of T. diversifolia and V. lasiopus was determined using GC-MS. Laboratory-based experiments were carried out using area preference method to assess the efficacy of the extracts against weevils for a test period of 5 h. Six groups of experiments were set up with ten S. zeamais in each test: positive control (Actellic), negative control (solvent only), and four different experimental extract concentrations (25, 50, 75, and 100%). RESULTS: The results indicated that T. diversifolia and V. lasiopus leaf extracts possess potent repellency effect on weevils. All the extracts simply discouraged S. zeamais from the treated areas recording significantly good levels of repellent activities between 26 and 96%. Furthermore, the GC-MS analysis manifested the presence of bioactive compound in the extracts which are associated with the repellency effects. CONCLUSION: The study scientifically confirms the traditional use of the T. diversifolia and V. lasiopus and provides important platform for further study on the extracts as bioresource of botanical repellent.

Volatile Compounds of Endophytic Bacillus spp. have Biocontrol Activity Against Sclerotinia sclerotiorum.

To develop an effective biological agent to control Sclerotinia sclerotiorum, three endophytic Bacillus spp. strains with high antagonistic activity were isolated from maize seed and characterized. In vitro assays revealed that the Bacillus endophytes could produce volatile organic compounds (VOC) that reduced sclerotial production and inhibited mycelial growth of S. sclerotiorum. Gas chromatography-mass spectrometry revealed that the selected strains produced 16 detectable VOC. Eight of the produced VOC exhibited negative effects on S. sclerotiorum, while a further four induced accumulation of reactive oxygen species in mycelial cells. A mixture of VOC produced by Bacillus velezensis VM11 caused morphological changes in the ultrastructure and organelle membranes of S. sclerotiorum mycelial cells. The bromophenol blue assay revealed a yellow color of untreated fungal mycelium, which grew faster and deeper from 24 to 72 h postinoculation, as an indication of reduced pH. The potassium permanganate (KMnO4) titration assay showed that the rate of oxalic acid accumulation was higher in minimal salt liquid medium cultures inoculated with untreated fungal plugs compared with the Bacillus VOC-treated ones. Interestingly, biological control assays using host-plant leaves challenged with treated fungal mycelial plugs produced reduced lesions compared with the control. These findings provide new viable possibilities of controlling diseases caused by S. sclerotiorum using VOC produced by Bacillus endophytes.

A scoping review on tsetse fly blood meal sources and its assay methods since 1956 to 2022.

BACKGROUND: Tsetse flies (Glossina spp.) are the definitive biological vectors of African trypanosomes in humans and animals. Controlling this vector is the most promising method of preventing trypanosome transmission. This requires a comprehensive understanding of tsetse biology and host preference to inform targeted design and management strategies, such as the use of olfaction and visual cues in tsetse traps. No current review exists on host preference and blood meal analyses of tsetse flies. METHODS: This review presents a meta-analysis of tsetse fly blood meal sources and the methodologies used to identify animal hosts from 1956 to August 2022. The Preferred Reporting Items for Systematic Reviews and Meta-Analyses extension for Scoping Reviews (PRIMA-ScR) was applied. This focused on tsetse-endemic countries, blood meal analysis methodologies and the blood meal hosts identified. The articles were retrieved and screened from databases using predetermined eligibility criteria. RESULTS: Only 49/393 of the articles retrieved matched the inclusion criteria. Glossina's main hosts in the wild included the bushbuck, buffalo, elephant, warthog, bushpig and hippopotamus. Pigs, livestock and humans were key hosts at the domestic interface. The least studied species included Glossina fuscipleuris, G. fusca, G. medicorum, G. tabaniformis and G. austeni. In the absence of preferred hosts, Glossina fed opportunistically on a variety of hosts. Precipitin, haemagglutination, disc diffusion, complement fixation, ELISA and PCR-based assays were used to evaluate blood meals. Cytochrome b (Cyt b) was the main target gene in PCR to identify the vertebrate hosts. CONCLUSIONS: Tsetse blood meal sources have likely expanded because of ecological changes that could have rendered preferred hosts unavailable. The major approaches for analysing tsetse fly blood meal hosts targeted Cyt b gene for species identification by Sanger sequencing. However, small-fragment DNAs, such as the mammalian 12S and 16S rRNA genes, along with second- and third-generation sequencing techniques, could increase sensitivity for host identification in multiple host feeders that Sanger sequencing may misidentify as "noise". This review of tsetse fly blood meal sources and approaches to host identification could inform strategies for tsetse control.

The Anopheles coluzzii range extends into Kenya: Detection, insecticide resistance profiles and population genetic structure in relation to conspecific populations in West and Central Africa.

Background: Anopheles coluzzii is a primary vector of malaria found in West and Central Africa, but its presence has hitherto never been documented in Kenya. A thorough understanding of vector bionomics is important as it enables the implementation of targeted and effective vector control interventions. Malaria vector surveillance efforts in the country have tended to focus on historically known primary vectors. In the current study, we sought to determine the taxonomic status of samples collected from five different malaria epidemiological zones in Kenya as well asdescribe the population genetic structure and insecticide resistance profiles in relation to other An. coluzzi populations. Methods: Mosquitoes were sampled as larvae from Busia, Kwale, Turkana, Kirinyaga and Kiambu counties, representing the range of malaria endemicities in Kenya, in 2019 and 2021 and emergent adults analysed using Whole Genome Sequencing data processed in accordance with the Anopheles gambiae 1000 Genomes Project phase 3. Where available, historical samples from the same sites were included for WGS. Results: This study reports the detection of Anopheles coluzzii for the first time in Kenya. The species was detected in Turkana County across all three time points sampled and its presence confirmed through taxonomic analysis. Additionally, we found a lack of strong population genetic differentiation between An. coluzzii from Kenya and those from the more northerly regions of West and Central Africa, suggesting they represent a connected extension to the known species range. Mutations associated with target-site resistance to DDT and pyrethroids and metabolic resistance to DDT were found at high frequencies of ~60%. The profile and frequencies of the variants observed were similar to An. coluzzii from West and Central Africa but the ace-1 mutation linked to organophosphate and carbamate resistance present in An. coluzzii from coastal West Africa was absent in Kenya. Conclusions: These findings emphasise the need for the incorporation of genomics in comprehensive and routine vector surveillance to inform on the range of malaria vector species, and their insecticide resistance status to inform the choice of effective vector control approaches.

Influence of soil fertility management technologies on phosphorus fractions, sorption characteristics, and use efficiency in humic Nitisols of Upper Eastern Kenya.

Fractions of phosphorus (P) and its sorption characteristics are affected by different soil fertility (FM) technologies which ultimately affect crop growth and productivity. However, the response of P fractions and sorption characteristics to soil fertility technologies that integrate diverse amendments is still poorly understood in acidic Nitisols. A randomized complete block design was layout in an acidic Nitisol to determine fractions of P, its sorption characteristics and use efficiencies in acidic Nitisols under various FM technologies in field conditions. The use of minimum tillage + maize residue + inorganic fertilizer + goat manure (MTCrGF) had the highest impact on and significantly increased resin-Pi, NaHCO3-Pi, and maximum P sorption (Smax) by 182, 76, and 52 mg P kg-1. Moreover, NaOH-Pi and Smax concentrations were higher under conventional tillage + maize residue + inorganic fertilizer + goat manure (CTCrGF) by 216 mg P kg-1 and 49 mg P kg-1 than the control. MTCrGF and CTCrGF also had the lowest P bonding energy (0.04 L mg-1). CTCrGF had the highest P partial productivity factor (0.093 and 0.140 kg biomass kg-1 P) and P agronomic efficiency (0.080 and 0.073 kg biomass kg-1 P) during the two cropping seasons. The results demonstrate the positive influence of combining multiple P sources on soil P fractions, sorption characteristics, and use efficiencies. Notably, combining either conventional or minimum tillage with maize straw and applying integrated manure and inorganic fertilizer (MTCrGF or CTCrGF) can increase the labile P concentrations and reduce the potential depletion of the non-renewable rock phosphate and the use of inorganic phosphatic fertilizers for agricultural production.

Farmers' knowledge, perceptions, and practices on animal trypanosomosis and the tsetse fly vector: A cross-sectional study around Kenya's Arabuko-Sokoke Forest Reserve at the livestock-wildlife interface.

Background: Animal African trypanosomosis (AAT) is a veterinary disease caused by trypanosomes transmitted cyclically by tsetse flies. AAT causes huge agricultural losses in sub-Saharan Africa. Both tsetse flies and trypanosomosis (T&T) are endemic in the study area inhabited by smallholder livestock farmers at the livestock-wildlife interface around Arabuko-Sokoke Forest Reserve (ASFR) in Kilifi County on the Kenyan coast. We assessed farmers' knowledge, perceptions and control practices towards T&T. Methods: A cross-sectional study was conducted during November and December 2017 to collect data from 404 randomly selected cattle-rearing households using a structured questionnaire. Descriptive statistics were used to determine farmers' knowledge, perceptions, and control practices towards T&T. Demographic factors associated with knowledge of T&T were assessed using a logistic regression model. Results: Participants consisted of 53% female, 77% married, 30% elderly (>55 years), and the majority (81%) had attained primary education or below. Most small-scale farmers (98%) knew the tsetse fly by its local name, and 76% could describe the morphology of the adult tsetse fly by size in comparison to the housefly's ( Musca domestica). Only 16% of the farmers knew tsetse flies as vectors of livestock diseases. Higher chances of adequate knowledge on T&T were associated with the participants' (i) age of 15-24 years (aOR 2.88 (95% CI 1.10-7.52), (ii) level of education including secondary (aOR 2.46 (95% CI 1.43-4.24)) and tertiary (aOR 3.80 (95% CI 1.54-9.37)), and (iii) employment status: self-employed farmers (aOR 6.54 (95% CI 4.36-9.80)). Conclusions: Our findings suggest that small-scale farmers around ASFR have limited knowledge of T&T. It is envisaged that efforts geared towards training of the farmers would bridge this knowledge gap and sharpen the perceptions and disease control tactics to contribute to the prevention and control of T&T.

Comparison of volatile blends and gene sequences of two isolates of Metarhizium anisopliae of different virulence and repellency toward the termite Macrotermes michaelseni.

Previously, we reported an interesting relationship between virulence and repellency of different isolates of the fungus Metarhizium anisopliae towards the termite Macrotermes michaelseni: the higher the virulence of a given isolate, the greater its repellency. In the present study, we compared the volatile profiles of two isolates, one that was more virulent (and repellent) and one that was less virulent (and repellent) to the termite. The prominent components of the two blends were characterized by gas chromatography-mass spectrometry and authenticated by gas chromatography co-injections with synthetic standards. There were both qualitative and quantitative differences between the two blends. The repellencies of synthetic blends of 10 prominent constituents of the volatiles of the two isolates were compared and that of the more virulent isolate was found to be significantly more repellent. Subtractive bioassays were carried out with one of the constituents of each of the two 10-component blends missing at a time to determine its relative contribution to the overall repellency. The results indicated that the repellency of the volatiles of each isolate was primarily due to synergistic effects of a smaller number of constituents. Intraspecific differences between the two isolates were also reflected in their nucleotide sequences.

Improved Sample Selection and Preparation Methods for Sampling Plans Used to Facilitate Rapid and Reliable Estimation of Aflatoxin in Chicken Feed.

Aflatoxin B1 (AFB1), a toxic fungal metabolite associated with human and animal diseases, is a natural contaminant encountered in agricultural commodities, food and feed. Heterogeneity of AFB1 makes risk estimation a challenge. To overcome this, novel sample selection, preparation and extraction steps were designed for representative sampling of chicken feed. Accuracy, precision, limits of detection and quantification, linearity, robustness and ruggedness were used as performance criteria to validate this modification and Horwitz function for evaluating precision. A modified sampling protocol that ensured representativeness is documented, including sample selection, sampling tools, random procedures, minimum size of field-collected aggregate samples (primary sampling), procedures for mass reduction to 2 kg laboratory (secondary sampling), 25 g test portion (tertiary sampling) and 1.3 g analytical samples (quaternary sampling). The improved coning and quartering procedure described herein (for secondary and tertiary sampling) has acceptable precision, with a Horwitz ratio (HorRat = 0.3) suitable for splitting of 25 g feed aliquots from laboratory samples (tertiary sampling). The water slurring innovation (quaternary sampling) increased aflatoxin extraction efficiency to 95.1% through reduction of both bias (-4.95) and variability of recovery (1.2-1.4) and improved both intra-laboratory precision (HorRat = 1.2-1.5) and within-laboratory reproducibility (HorRat = 0.9-1.3). Optimal extraction conditions are documented. The improved procedure showed satisfactory performance, good field applicability and reduced sample analysis turnaround time.

Comparison of quantitative polymerase chain reaction, Kato-Katz and circulating cathodic antigen rapid test for the diagnosis of Schistosoma mansoni infection: A cross-sectional study in Kirinyaga County, Kenya.

The current standard diagnostic tests for Schistosoma mansoni are the Kato-Katz and circulating cathodic antigen (CCA) techniques. However, these techniques have been documented to have several limitations that have a direct impact on schistosomiasis control programmes. Therefore, there is a need for more sensitive and specific tests for diagnosing schistosomiasis. This study compared the performance of quantitative polymerase chain reaction (qPCR), Kato-Katz, and point-of-care circulating cathodic antigen (POC-CCA) techniques in the diagnosis of S. mansoni infection in the Mwea irrigation scheme, Kirinyaga County in Central Kenya. We carried out a cross-sectional study on 357 individuals residing in four villages in the Mwea irrigation scheme. The participants provided urine and stool samples which were screened for S. mansoni infections using the three techniques. The prevalence of S. mansoni by each technique was calculated and 95% confidence intervals estimated using binomial regression model. Sensitivity and specificity were determined using 2 × 2 contingency tables and compared using the McNemar's chi-square test. Positive and negative predictive values were also determined using the weighted generalized score chi-square test for paired data. The study showed that the prevalence of S. mansoni was 32.8%, 62.5% and 72.8% using Kato-Katz, POC-CCA and qPCR techniques, respectively. Further, when using Kato-Katz as a gold standard, POC-CCA sensitivity was 78.6% and specificity was 45.4%, while qPCR sensitivity was 97.4% and specificity was 39.2%. When using qPCR as the gold standard, Kato-Katz sensitivity was 43.8% and specificity was 96.9%, while POC-CCA sensitivity was 78.1% and specificity was 79.4%. Finally, when using the averaged results from the three techniques as the gold standard, the sensitivity was 41.6%, 79.4% and 92.5% for Kato-Katz, POC-CCA and qPCR, respectively, with a specificity of 100% for all techniques. Kato-Katz technique showed low sensitivity compared to the POC-CCA and qPCR despite it being the most commonly preferred method of choice to diagnose S. mansoni infections. qPCR showed superior sensitivity followed by POC-CCA, hence it can be used as an alternative or to confirm the results obtained by the Kato-Katz technique.

Effects of increased plastic film residues on soil properties and crop productivity in agro-ecosystem.

Intensive use of low-density polyethylene (LDPE) plastic films in agro-ecosystems has raised considerable concerns due to the increasing film residues in soils. It is unclear how the increased film residues affect soil properties and crop productivity and whether biodegradable (Bio) film can substitute LDPE. To address the issue, we designed a landfill experiment with different addition levels of plastic residue into soils of maize (Zea mays L.) field from 2018 to 2019. Six treatments were arranged as PMT1-T3/BioT1-T3, representing the low, medium, and high-level application of LDPE / Bio film fragments, with no residual film, applied as CK. Results show that, soil bulk density was significantly increased from 1.19 to 1.31 g/cm3 regardless of residue types. In contrast, soil porosity was lowered from 58.03% in CK to 57.36% in Bio and 56.12% in LDPE significantly (P < 0.05). Increased residues improved soil nitrogen level and lowered the C/N ratio significantly. Also, it decreased microbial biomass C and N levels but with no change in C/N (P < 0.05). Maize yield and WUE decreased, while soil water storage increased significantly. LDPE residues affected soil properties and productivity partly lower than Bio ones did, but the negative effects of them were similar in the maize field.

Horizontal Transmission of the Symbiont Microsporidia MB in Anopheles arabiensis.

The recently discovered Anopheles symbiont, Microsporidia MB, has a strong malaria transmission-blocking phenotype in Anopheles arabiensis, the predominant Anopheles gambiae species complex member in many active transmission areas in eastern Africa. The ability of Microsporidia MB to block Plasmodium transmission together with vertical transmission and avirulence makes it a candidate for the development of a symbiont-based malaria transmission blocking strategy. We investigate the characteristics and efficiencies of Microsporidia MB transmission between An. arabiensis mosquitoes. We show that Microsporidia MB is not transmitted between larvae but is effectively transmitted horizontally between adult mosquitoes. Notably, Microsporidia MB was only found to be transmitted between male and female An. arabiensis, suggesting sexual horizontal transmission. In addition, Microsporidia MB cells were observed infecting the An. arabiensis ejaculatory duct. Female An. arabiensis that acquire Microsporidia MB horizontally are able to transmit the symbiont vertically to their offspring. We also investigate the possibility that Microsporidia MB can infect alternate hosts that live in the same habitats as their An. arabiensis hosts, but find no other non-anopheline hosts. Notably, Microsporidia MB infections were found in another primary malaria African vector, Anopheles funestus s.s. The finding that Microsporidia MB can be transmitted horizontally is relevant for the development of dissemination strategies to control malaria that are based on the targeted release of Microsporidia MB infected Anopheles mosquitoes.

Dual plastic film and straw mulching boosts wheat productivity and soil quality under the El Nino in semiarid Kenya.

The extreme climate events such as El Nino seriously threaten crop production and agro-ecological sustainability because of the aggravated environmental stresses worldwide, particularly in sub-Saharan Africa. To address this issue, we investigated the effects of dual plastic film and straw mulching in ridge-furrow (RF) system on wheat productivity, soil carbon and nitrogen stocks in a semiarid area in Kenya from 2015 to 2017. The experimental site represents a typical semiarid continental monsoon climate, and soil type is chromic vertisols. Field experiment with randomized block design consisted of six RF treatments as follows: 1) dual black plastic film and straw mulching (RFbS), 2) dual transparent plastic film and straw mulching (RFtS), 3) sole black plastic film mulching (RFb), 4) sole transparent plastic mulching RF (RFt), 5) sole straw mulching (RFS) and 6) no mulching (CK). The results indicated that seasonal dynamics of rainfall and air temperature fit in with the weather type of El Nino over four growing seasons. RFbS, RFtS, RFb and RFt significantly increased soil water storage (SWS), topsoil temperature, aboveground biomass, grain yield and water use efficiency across four growing seasons (p < 0.05) as compared with CK. Among all the treatments, RFbS and RFtS achieved the greatest SWS, AgB, grain yield and WUE, owing to improved soil hydro-thermal status in both treatments. Critically, RFbS and RFtS significantly improved soil organic carbon and total nitrogen, soil bulk density and the C:N ratio following four growing seasons, comparing with other treatments (p < 0.05). Besides, RFbS and RFtS gave the highest economic returns among all treatments. For the first time, we found that dual plastic film and straw mulching could serve as a sustainable land management to boost wheat productivity and improve soil quality under El Nino in semiarid areas of SSA.

The expression of Spodoptera exigua P450 and UGT genes: tissue specificity and response to insecticides.

Cytochrome P450 and UDP-glucosyltransferase (UGT) as phase I and phase II metabolism enzymes, respectively, play vital roles in the breakdown of endobiotics and xenobiotics. Insects can increase the expression of detoxification enzymes to cope with the stress from xenobiotics including insecticides. However, the molecular mechanisms for insecticide detoxification in Spodoptera exigua remain elusive, and the genes conferring insecticide metabolisms in this species are less well reported. In this study, 68 P450 and 32 UGT genes were identified. Phylogenetic analysis showed gene expansions in CYP3 and CYP4 clans of P450 genes and UGT33 family of this pest. P450 and UGT genes exhibited specific tissue expression patterns. Insecticide treatments in fat body cells of S. exigua revealed that the expression levels of P450 and UGT genes were significantly influenced by challenges of abamectin, lambda-cyhalothrin, chlorantraniliprole, metaflumizone and indoxacarb. Multiple genes for detoxification were affected in expression levels after insecticide exposures. The results demonstrated that lambda-cyhalothrin, chlorantraniliprole, metaflumizone and indoxacarb induced similar responses in the expression of P450 and UGT genes in fat body cells; eight P450 genes and four UGT genes were co-up-regulated significantly, and no or only a few CYP/UGT genes were down-regulated significantly by these four insecticides. However, abamectin triggered a distinct response for P450 and UGT gene expression; more P450 and UGT genes were down-regulated by abamectin than by the other four compounds. In conclusion, P450 and UGT genes from S. exigua were identified, and different responses to abamectin suggest a different mechanism for insecticide detoxification.

UV Resistance of bacteria from the Kenyan Marine cyanobacterium Moorea producens.

UV resistance of bacteria isolated from the marine cyanobacterium Moorea producens has not been observed previously, findings which highlight how unsafe germicidal UV irradiation for sterilization of air, food, and water could be. Further, UV resistance of Bacillus licheniformis is being observed for the first time. This study focused on bacteria isolated from the marine cyanobacterium M. producens collected off the Kenyan coast at Shimoni, Wasini, Kilifi, and Mida. UV irradiance of isolates (302 nm, 70 W/m2 , 0-1 hr) established B. licheniformis as the most UV resistant strain, with the following order of taxon resistance: Bacilli> Î³ proteobacteria > Actinobacteria. UV resistance was independent of pigmentation. The maximum likelihood phylogenetic distance determined for both B. licheniformis and Bacillus aerius relative to M. producens CCAP 1446/4 was 2.0. Survival of B. licheniformis upon UV irradiance followed first-order kinetics (k = 0.035/min, R2  = 0.88). Addition of aqueous extracts (2, 10, 20 and 40 mg/ml) of this B. licheniformis strain on the less resistant Marinobacterium stanieri was not significant, however, the commercial sunscreen benzophenone-3 (BP-3) positive control and the time of irradiance were significant. Detection of bacteria on M. producens filaments stained with acridine orange confirmed its nonaxenic nature. Although the chemistry of UV resistance in cyanobacteria has been studied in depth revealing for example the role of mycosporine like amino acids (MAAs) in UV resistance less is known about how bacteria resist UV irradiation. This is of interest since cyanobacteria live in association with bacteria.

Transcription factors CncC/Maf and AhR/ARNT coordinately regulate the expression of multiple GSTs conferring resistance to chlorpyrifos and cypermethrin in Spodoptera exigua.

BACKGROUND: Glutathione S-transferases (GSTs) are a superfamily of multifunctional dimeric proteins existing in both prokaryotic and eukaryotic organisms. They are involved in the detoxification of both endogenous and exogenous electrophiles, including insecticides. However, the molecular mechanisms underlying the regulation of GST genes in insects are poorly understood. RESULTS: We first identified at least three GST genes involved in resistance to the insecticides chlorpyrifos and cypermethrin. Analysis of upstream sequences revealed that three GSTs (SeGSTo2, SeGSTe6 and SeGSTd3) harbor the same cap 'n' collar C/muscle aponeurosis fibromatosis (CncC/Maf) binding site, and SeGSTo2 and SeGSTe6 contain the aryl hydrocarbon receptor/aryl hydrocarbon receptor nuclear translocator (AhR/ARNT) binding site. Luciferase reporter assay showed co-transfection of reporter plasmid containing the SeGSTe6 promoter with CncC and/or Maf expressing constructs significantly boosted transcription. Similarly, AhR and/or ARNT expressing constructs also significantly increased the promoter activities. The co-transfection of mutated reporter plasmid with CncC/Maf or AhR/ARNT did not increase transcription activity anymore. Constitutive over-expression of CncC, Maf and AhR was also found in the HZ16 strain, which might be the molecular mechanism for up-regulated expression of multiple detoxification genes conferring resistance to insecticides. CONCLUSION: These results suggest that CncC/Maf and AhR/ARNT coordinately regulate the expression of multiple GST genes involved in insecticide resistance in Spodoptera exigua. © 2019 Society of Chemical Industry.

Effects of methimazole on Drosophila glucolipid metabolism in vitro and in vivo.

Methimazole (MMI) is an antithyroid agent widely used in the treatment of hyperthyroidism, and metabolized by cytochrome P450 enzymes and flavin-containing monooxygenases in mammals. However, drug overdose and the inadequate detoxification of the metabolite(s) are responsible for hepatocellular damage and organ dysfunction. Depending on the desired properties, Drosophila melanogaster has recently emerged as an ideal model organism for the study of human diseases. Here we investigated the changes in metabolic profiles and mRNA expressions related to glucolipid metabolism in response to treatment with MMI in Drosophila. Remarkable loss of lifespan occurred in fruit flies fed on the diets containing 10 or 30mM MMI compared to unsupplemented controls. To examine whether MMI affects glucolipid metabolism in vitro and in vivo, fruit flies were fed diets containing 30mM MMI for two weeks and Drosophila S2 cells were incubated with 300µM MMI for 48h. Measurements of metabolites showed that triglyceride content dramatically decreased (30.56% in vivo and 18.13% in vitro), and glycogen content significantly increased (10.7% in vivo and 126.8% in vitro). Quantitative analyses indicated that mRNA expression levels of Dmfmo1, s6k, dilp2, acc and dilp5 genes involved in metabolic homeostasis were remarkably down-regulated in vivo and in vitro. Meanwhile, the addition of MMI could significantly reduce the lipid droplet content in S2 cells by approximately 25% compared to control subjects. These data may provide a biological basis for the study of MMI on disease symptoms and complications, and discovery of therapeutic treatments.

Molecular detection and phylogenetic analysis of Kenyan human bocavirus isolates.

INTRODUCTION: The commonly expected causative agents associated with flu-like symptoms in Kenya are the classical viral pathogens identifiable as influenza virus, adenovirus, parainfluenza virus, enteroviruses, respiratory syncytial virus (RSV) and rhinovirus. However, newer agents have been identified globally that present with illnesses clinically indistinguishable from those caused by the classical pathogens; one of them is human bocavirus. METHODOLOGY: A total of 384 specimens were analyzed, primarily to determine if the emerging human bocavirus (HBoV) infections exist in Kenya as coinfections with other respiratory viruses and to describe the genotype of the virus in circulation. In brief, viral nucleic acids were extracted from culture supernatants, amplified by PCR, and sequenced. RESULTS: HBoV DNA was amplified from 1.8% of screened specimens. Coinfection with parainfluenza virus, adenovirus, and enterovirus was 2.5%, 2%, and 1.4%, respectively. Multiple coinfections consisting of HBoV plus two other viruses were found in 3% of specimens. Isolation occurred in the months of January, March, April, August, and November. Retrospective review of clinical parameters indicated that all the individuals complained of non-specific symptoms, mainly fever, coughs, nasal stuffiness, runny noses, and vomiting. Phylogenetically, the GenBank deposited sequences of this study's isolates cluster closely to the reference strain NC_07455 (HBoV1). CONCLUSION: Coinfections with human bocavirus (HBoV1) occur in Kenya, and high incidence might primarily be during the early stages of children's lives.

No apparent reduction in schistosome burden or genetic diversity following four years of school-based mass drug administration in mwea, central kenya, a heavy transmission area.

BACKGROUND: Schistosomiasis is a debilitating neglected tropical disease that infects over 200 million people worldwide. To combat this disease, in 2012, the World Health Organization announced a goal of reducing and eliminating transmission of schistosomes. Current control focuses primarily on mass drug administration (MDA). Therefore, we monitored transmission of Schistosoma mansoni via fecal egg counts and genetic markers in a typical school based MDA setting to ascertain the actual impacts of MDA on the targeted schistosome population. METHODS: For 4 years, we followed 67 children enrolled in a MDA program in Kenya. Infection status and egg counts were measured each year prior to treatment. For 15 of these children, for which there was no evidence of acquired resistance, meaning they became re-infected following each treatment, we collected microsatellite genotype data from schistosomes passed in fecal samples as a representation of the force of transmission between drug treatments. We genotyped a total of 4938 parasites from these children, with an average of 329.2 parasites per child for the entire study, and an average of 82.3 parasites per child per annual examination. We compared prevalence, egg counts, and genetic measures including allelic richness, gene diversity (expected heterozygosity), adult worm burdens and effective number of breeders among time points to search for evidence for a change in transmission or schistosome populations during the MDA program. FINDINGS: We found no evidence of reduced transmission or schistosome population decline over the course of the program. Although prevalence declined in the 67 children as it did in the overall program, reinfection rates were high, and for the 15 children studied in detail, schistosome egg counts and estimated adult worm burdens did not decline between years 1 and 4, and genetic diversity increased over the course of drug treatment. INTERPRETATION: School based control programs undoubtedly improve the health of individuals; however, our data show that in an endemic area, such a program has had no obvious effect on reducing transmission or of significantly impacting the schistosome population as sampled by the children we studied in depth. Results like these, in combination with other sources of information, suggest more integrated approaches for interrupting transmission and significantly diminishing schistosome populations will be required to achieve sustainable control.