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The purpose of the study is to examine the role and impacts of growth hormone in maximizing animal products. Growth hormones are biological stimulants that are found either naturally in the organism or synthetically manufactured. Phytoestrogens, phytoprogestrons and Phenolic compounds are hormones from plants. Drugs from of placenta and colostrums of cow`s contain progesterone, estrogene, gonadotropin, and prostaglandins hormones. Growth hormones have got popular applications in dairy, beef, feed improvement and Biopharmaceutical productions with the aim of producing valuable products: fat free meat (Porcine Somatotropin hormone in pigs), nutritionally and medicinally reach milk (Bovine Somatotropin hormone in cattle), palatable and disease and insect pest resistant forage crop production. They have got also contribution in maximizing livestock production by involving in adjusting animals’ reproductive process such as oestrus synchronization and superovulation mainly during artificial insemination and embryo transfer. Controversially, these hormones have wide impacts on human being, animal welfare, environment and etc. Contamination of ground water by hormones that are found in the animals` excreta will cause deleterious effects such as cancer, loss of fertility, and some imbalance of minerals in the water and soil.

Microbial genes and their product were diverse and beneficial for heavy metal bioremediation from the contaminated sites. Screening of genes and gene products plays a significant role in the detoxification of pollutants. Understanding of the promoter region and its regulatory elements is a vital implication of microbial genes. To the best of our knowledge, there is no in silico study reported so far on mer gene families used for heavy metal bioremediation. The motif distribution was observed densely upstream of the TSSs (transcription start sites) between +1 and -350 bp and sparsely distributed above -350 bp, according to the current study. MEME identified the best common candidate motifs of TFs (transcription factors) binding with the lowest e value (7.2e-033) and is the most statistically significant candidate motif. The EXPREG output of the 11 TFs with varying degrees of function such as activation, repression, transcription, and dual purposes was thoroughly examined. Data revealed that transcriptional gene regulation in terms of activation and repression was observed at 36.4% and 54.56%, respectively. This shows that most TFs are involved in transcription gene repression rather than activation. Likewise, EXPREG output revealed that transcriptional conformational modes, such as monomers, dimers, tetramers, and other factors, were also analyzed. The data indicated that most of the transcriptional conformation mode was dual, which accounts for 96%. CpG island analysis using online and offline tools revealed that the gene body had fewer CpG islands compared to the promoter regions. Understanding the common candidate motifs, transcriptional factors, and regulatory elements of the mer operon gene cluster using a machine learning approach could help us better understand gene expression patterns in heavy metal bioremediation.

The leather industry is one of the major polluting industries in Ethiopia. Environment protection has prompted industrialists and scientists to re-look at transformation of tannery solid waste materials to valuable products using enzymatic hydrolysis. Acid and liming process of gelatin production generate large amount of wastes and used limited raw materials. This study was therefore, aimed to extract papain from papaya fruit latex and use it for transforming raw trimming to gelatin. Extraction and purification was carried out using Three Phase Partitioning. Protein content and protease activity were determined using lowry and casein hydrolysis method, respectively. Effect of agitation time taken, crude extract to t-butanol ratio, ammonium sulphate concentration and temperature on purification fold and activity recovery of papain was evaluated and process was optimized. At its optimized parameters, the intermediate phase of TPP showed 94.14% percentage of protease activity recovery and 8.31 purification fold. At its optimized parameters, bottom phase showed 75.15% percentage of protease activity recovery and 6.61 purification fold. Enzymatic hydrolysis of raw trimming produced 68 wt% and 64 wt% gelatin recovery from the use of crude and intermediate phase papain, respectively. Relatively low gel strength of gelatin with bloom value of 95g and 87g were obtained from hydrolysis of raw trimming by intermediate and crude papain, respectively. This study showed that the use of crude extract for transforming raw trimming to gelatin was found good as it showed comparable protease activity. The TPP can be employed for the extraction and purification of proteases from papaya fruit for industrial application that requires it in partial pure form.

Background: Breast cancer is the most common cancer-affecting women globally, with disproportionally high mortality rates in lower-income countries, including Ethiopia. The stage at diagnosis is a well-defined predictive system that determines the likelihood of breast cancer mortality. Early-stage breast cancer at diagnosis is associated with better treatment outcomes as compared with late stage. Although there are numerous primary studies on women with breast cancer with different proportions of early-stage breast cancer, there is currently no summary data on what proportion of breast cancer was diagnosed at early-stage in Ethiopia. This study focused on a pooled proportion of early-stage breast cancer at diagnosis in Ethiopia., Methods: By using key terms, Medline through Pub-Med, Google Scholar, Science Direct, HINARI and Medley were searched about breast cancer in Ethiopia, and a total of 288 articles were retrieved. After screening the articles and quality of each article was assessed using Newcastle-Ottawa Scale. Finally, 41 articles were used for the final pooled proportion. A random effects model was used to estimate the pooled prevalence and heterogeneity of included studies that were then assessed by using prediction interval., Results: Pooled proportion of early-stage breast cancer at diagnosis in Ethiopian hospitals was found to be 36% with a 95% confidence interval ranging from 31 to 41% and a 95% prediction interval ranging from 28 to 45%., Conclusion: Most breast cancer patients (64%) in Ethiopia are diagnosed at a late-stage. This contributes to the high mortality rates of breast cancer among women in Ethiopia. Therefore, in line with recommendations by the World Health Organization, we recommend that there should be an emphasis in Ethiopia to focus on early detection of breast cancer., (© 2024. The Author(s).)

Protists in general comprise about one-third of the parasitic species infecting arthropod vectors, the role of free-living and epibiotic ciliates on mosquitoes have been insufficiently studied either due to their low pathogenicity or facultative parasites. Studies have shown that exposure of Paramecium ciliate protists, like Vorticella species, to first instar Culex nigripalpus Theobald, larvae delayed larval development and reduced biomass of emerged adults due to competition for food sources like bacteria and other microbes essential to mosquito growth and survival. Thus, we report on the capacity of a Vorticella sp. protist's ability to cross-infect host species and parasitize multiple mosquito larvae. The unique adapted behavior with the ability to remain on the exuviae in tree hole habitats provide a novel delivery system to develop products for target species-specific mosquitocides, larvicides, or viricides to be applied and sustained in aquatic systems., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Inc.)

Background: Multi-month dispensing (MMD) of antiretroviral therapy (ART) is an integral component of differentiated HIV service delivery for people living with HIV (PLHIV). Although many countries have scaled up ART dispensing to 3-month intervals, Ethiopia was the first African country to implement six-month dispensing (6-MMD) at scale, introducing its Appointment Spacing Model (ASM) for people doing well on ART in 2017. As of June 2021, 51.4% (n = 215,101) of PLHIV on ART aged ≥ 15 years had enrolled in ASM. Since little is known about the benefits and challenges of ASM perceived by Ethiopian clients and their healthcare workers (HCWs), we explored how the ASM was being implemented in Ethiopia's Oromia region in September 2019., Methods: Using a parallel convergent mixed-methods study design, we conducted 6 focus groups with ASM-eligible enrolled clients, 6 with ASM-eligible non-enrolled clients, and 22 in-depth interviews with HCWs. Data were audio-recorded, transcribed and translated into English. We used thematic analysis, initially coding deductively, followed by inductive coding of themes that emerged from the data, and compared the perspectives of ASM-enrolled and non-enrolled clients and their HCWs., Results: Participants enrolled in ASM and HCWs perceived client-level ASM benefits to include time and cost-savings, fewer work disruptions, reduced stigma due to fewer clinic visits, better medication adherence and improved overall health. Perceived health system-level benefits included improved quality of care, decongested facilities, reduced provider workloads, and improved record-keeping. Although non-enrolled participants anticipated many of the same benefits, their reasons for non-enrollment included medication storage challenges, concerns over less frequent health monitoring, and increased stress due to the large quantities of medicines dispensed. Enrolled participants and HCWs identified similar challenges, including client misunderstandings about ASM and initial ART stock-outs., Conclusions: ASM with 6-MMD was perceived to have marked benefits for clients and health systems. Clients enrolled in the ASM and their HCWs had positive experiences with the model, including perceived improvements in efficiency, quality and convenience of HIV treatment services. The concerns of non-ASM enrolled participants suggest the need for enhanced client education about the model and more discreet and efficiently packaged ART and highlight that ASM is not ideal for all clients., (© 2023. The Author(s).)

Among disease vectors, Aedes aegypti (L.) (Diptera: Culicidae) is one of the most insidious species in the world. The disease burden created by this species has dramatically increased in the past 50 years, and during this time countries have relied on pesticides for control and prevention of viruses borne by Ae. aegypti. The small number of available insecticides with different modes of action had led to increases in insecticide resistance, thus, strategies, like the "Incompatible Insect Technique" using Wolbachia's cytoplasmic incompatibility are desirable. We evaluated the effect of releases of Wolbachia infected Ae. aegypti males on populations of wild Ae. aegypti in the metropolitan area of Houston, TX. Releases were conducted by the company MosquitoMate, Inc. To estimate mosquito population reduction, we used a mosquito abundance Bayesian hierarchical estimator that accounted for inefficient trapping. MosquitoMate previously reported a reduction of 78% for an intervention conducted in Miami, FL. In this experiment we found a reduction of 93% with 95% credibility intervals of 86% and 96% after six weeks of continual releases. A similar result was reported by Verily Life Sciences, 96% [94%, 97%], in releases made in Fresno, CA., Competing Interests: The authors have declared that no competing interests exist., (Copyright: This is an open access article, free of all copyright, and may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. The work is made available under the Creative Commons CC0 public domain dedication.)

Microbial genes and their product were diverse and beneficial for heavy metal bioremediation from the contaminated sites. Screening of genes and gene products plays a significant role in the detoxification of pollutants. Understanding of the promoter region and its regulatory elements is a vital implication of microbial genes. To the best of our knowledge, there is no in silico study reported so far on mer gene families used for heavy metal bioremediation. The motif distribution was observed densely upstream of the TSSs (transcription start sites) between +1 and -350 bp and sparsely distributed above -350 bp, according to the current study. MEME identified the best common candidate motifs of TFs (transcription factors) binding with the lowest e value (7.2 e -033) and is the most statistically significant candidate motif. The EXPREG output of the 11 TFs with varying degrees of function such as activation, repression, transcription, and dual purposes was thoroughly examined. Data revealed that transcriptional gene regulation in terms of activation and repression was observed at 36.4% and 54.56%, respectively. This shows that most TFs are involved in transcription gene repression rather than activation. Likewise, EXPREG output revealed that transcriptional conformational modes, such as monomers, dimers, tetramers, and other factors, were also analyzed. The data indicated that most of the transcriptional conformation mode was dual, which accounts for 96%. CpG island analysis using online and offline tools revealed that the gene body had fewer CpG islands compared to the promoter regions. Understanding the common candidate motifs, transcriptional factors, and regulatory elements of the mer operon gene cluster using a machine learning approach could help us better understand gene expression patterns in heavy metal bioremediation., Competing Interests: The authors declare that they have no competing interests., (Copyright © 2022 Duguma Dibbisa and Gobena Wagari.)

Background: Understanding the clinical features of COVID-19 and duration for resolution of symptoms is crucial for isolation of patients and tailoring public health messaging, interventions and policy. Therefore, this study aims to assess the median duration of COVID-19 signs and symptoms' resolution and explore its predictors among symptomatic COVID-19 patients in Ethiopia., Methods: A hospital-based prospective cohort study involving 124 COVID-19 cases was conducted at Eka Kotebe General Hospital, COVID-19 Isolation and Treatment Center. The study participants were all symptomatic COVID-19 adult patients admitted to the hospital from 18 March to 20 August 2020. Physicians at the centre recorded the data using a log sheet. Cox proportional-hazards regression model was conducted. Statistical significance was defined at P  < 0.05., Results: A total of 124 symptomatic COVID-19 patients with a mean age of 42 years (±17) were involved in the study. The median duration of symptom resolution of COVID-19 was seven days with a minimum of two and a maximum of sixty-eight days. Sex and body mass index (BMI) were statistically significant predictors of the symptom resolution. The hazard of having delayed sign or symptom resolution in males was 55% higher than in females ( P  = 0.039; CI: 0.22-0.96) and the hazard of delayed sign or symptom resolution in those with BMI ≥ 25 kg/m 2 was 35% higher than in those with BMI < 25 kg/m 2 ( P  = 0.041; CI: 0.44-0.98])., Conclusions: The median duration of COVID-19 symptom resolution was seven days. Being male and/or having a BMI ≥ 25 kg/m 2 were predictors of a delayed sign or symptom resolution time. Therefore, it is important to consider proportion of males and those with BMI ≥ 25 kg/m 2 when preparing isolation and treatment centres. Males and individuals with BMI ≥ 25 kg/m 2 shall also be given priority when shielding from the COVID-19., Competing Interests: The authors declare no conflict of interest., (© 2021 The Authors. Lifestyle Medicine published by John Wiley & Sons Ltd.)

Background: The coronavirus disease 2019 (COVID-19) is caused by severe acute respiratory syndrome coronavirus (SARS-CoV-2) and became pandemic after emerging in Wuhan, China, in December 2019. Several studies have been conducted to understand the key features of COVID-19 and its public health impact. However, the prognostic factors of COVID-19 are not well studied in the African setting. In this study, we aim to determine the epidemiological and clinical features of COVID-19 cases, immunological and virological courses, interaction with nutritional status, and response to treatment for COVID-19 patients in Ethiopia., Methods: A multi-center cohort study design will be performed. Patients with confirmed COVID-19 infection admitted to selected treatment centers will be enrolled irrespective of their symptoms and followed-up for 12 months. Baseline epidemiological, clinical, laboratory and imaging data will be collected from treatment records, interviews, physical measurements, and biological samples. Follow-up data collection involves treatment and prognostic outcomes to be measured using different biomarkers and clinical parameters. Data collection will be done electronically using the Open Data Kit (ODK) software package and then exported to STATA/SPSS for analysis. Both descriptive and multivariable analyses will be performed to assess the independent determinants of the treatment outcome and prognosis to generate relevant information for informed prevention and case management. The primary outcomes of this study are death/survival and viral shedding. Secondary outcomes include epidemiological characteristics, clinical features, genetic frequency shifts (genotypic variations), and nutritional status., Discussion: This is the first large prospective cohort study of patients in hospitals with COVID-19 in Ethiopia. The results will enable us to better understand the epidemiology of SARS-CoV-2 in Africa. This study will also provide useful information for effective public health measures and future pandemic preparedness and in response to outbreaks. It will also support policymakers in managing the epidemic based on scientific evidence., Trial Registration: The Protocol prospectively registered in ClinicalTrials.gov (NCT04584424) on 30 October, 2020., (© 2021. The Author(s).)

Objectives: Kangaroo Mother Care (KMC), prolonged skin-to-skin care of the low birth weight baby with the mother plus exclusive breastfeeding reduces neonatal mortality. Global KMC coverage is low. This study was conducted to develop and evaluate context-adapted implementation models to achieve improved coverage., Design: This study used mixed-methods applying implementation science to develop an adaptable strategy to improve implementation. Formative research informed the initial model which was refined in three iterative cycles. The models included three components: (1) maximising access to KMC-implementing facilities, (2) ensuring KMC initiation and maintenance in facilities and (3) supporting continuation at home postdischarge., Participants: 3804 infants of birth weight under 2000 g who survived the first 3 days, were available in the study area and whose mother resided in the study area., Main Outcome Measures: The primary outcomes were coverage of KMC during the 24 hours prior to discharge and at 7 days postdischarge., Results: Key barriers and solutions were identified for scaling up KMC. The resulting implementation model achieved high population-based coverage. KMC initiation reached 68%-86% of infants in Ethiopian sites and 87% in Indian sites. At discharge, KMC was provided to 68% of infants in Ethiopia and 55% in India. At 7 days postdischarge, KMC was provided to 53%-65% of infants in all sites, except Oromia (38%) and Karnataka (36%)., Conclusions: This study shows how high coverage of KMC can be achieved using context-adapted models based on implementation science. They were supported by government leadership, health workers' conviction that KMC is the standard of care, women's and families' acceptance of KMC, and changes in infrastructure, policy, skills and practice., Trial Registration Numbers: ISRCTN12286667; CTRI/2017/07/008988; NCT03098069; NCT03419416; NCT03506698., Competing Interests: Competing interests: None declared., (© Author(s) (or their employer(s)) 2021. Re-use permitted under CC BY. Published by BMJ.)

Mosquito-borne pathogens contribute significantly to the global burden of infectious diseases and are a continuing public health concern in the United States. Blood feeding by vector mosquitoes is a critical step in the transmission of human pathogens. Continuous surveillance of mosquito feeding patterns, especially in major population centers, is necessary for sustainable, effective control strategies. To better understand female feeding habits in Harris County, TX, we trapped mosquitoes from various locations, distributed among urban and semi-urban environments. Bloodmeal hosts were determined using a cytochrome C oxidase I DNA barcoding strategy. We identified a diverse array of vertebrate hosts with a high degree of avian host utilization, most surprisingly from anthropophilic species like Aedes aegypti (L.). We also detected sequences from two different vertebrate hosts in about half of specimens examined, suggesting that multiple bloodmeals had been acquired in the same feeding cycle by a sizable fraction of females in both urban and semi-urban locations. The high proportion of feeding on domestic chickens may indicate that a significant number of homeowners are rearing chickens within close proximity to study trap sites. As non-amplifying hosts, chickens may have a diluting effect on West Nile virus, as well as a zooprophylactic effect in their immediate vicinities. Ultimately, spatial and temporal host utilization patterns add insight into potential disease transmission dynamics, thereby informing vector control strategies in Harris County and other metropolitan areas., (© The Author(s) 2020. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Introduction: Kangaroo Mother Care (KMC) is the practice of early, continuous and prolonged skin-to-skin contact between the mother and the baby with exclusive breastfeeding. Despite clear evidence of impact in improving survival and health outcomes among low birth weight infants, KMC coverage has remained low and implementation has been limited. Consequently, only a small fraction of newborns that could benefit from KMC receive it., Methods and Analysis: This implementation research project aims to develop and evaluate district-level models for scaling up KMC in India and Ethiopia that can achieve high population coverage. The project includes formative research to identify barriers and contextual factors that affect implementation and utilisation of KMC and design scalable models to deliver KMC across the facility-community continuum. This will be followed by implementation and evaluation of these models in routine care settings, in an iterative fashion, with the aim of reaching a successful model for wider district, state and national-level scale-up. Implementation actions would happen at three levels: 'pre-KMC facility'-to maximise the number of newborns getting to a facility that provides KMC; 'KMC facility'-for initiation and maintenance of KMC; and 'post-KMC facility'-for continuation of KMC at home. Stable infants with birth weight<2000 g and born in the catchment population of the study KMC facilities would form the eligible population. The primary outcome will be coverage of KMC in the preceding 24 hours and will be measured at discharge from the KMC facility and 7 days after hospital discharge., Ethics and Dissemination: Ethics approval was obtained in all the project sites, and centrally by the Research Ethics Review Committee at the WHO. Results of the project will be submitted to a peer-reviewed journal for publication, in addition to national and global level dissemination., Study Status: WHO approved protocol: V.4-12 May 2016-Protocol ID: ERC 2716. Study implementation beginning: April 2017. Study end: expected March 2019., Trial Registration Number: Community Empowerment Laboratory, Uttar Pradesh, India (ISRCTN12286667); St John's National Academy of Health Sciences, Bangalore, India and Karnataka Health Promotion Trust, Bangalore, India (CTRI/2017/07/008988); Society for Applied Studies, Delhi (NCT03098069); Oromia, Ethiopia (NCT03419416); Amhara, SNNPR and Tigray, Ethiopia (NCT03506698)., Competing Interests: Competing interests: None declared., (© Author(s) (or their employer(s)) 2019. Re-use permitted under CC BY. Published by BMJ.)

The mosquitoes Aedes aegypti (L.) and Ae .  albopictus Skuse are the major vectors of dengue, Zika, yellow fever, and chikungunya viruses worldwide. Wolbachia , an endosymbiotic bacterium present in many insects, is being utilized in novel vector control strategies to manipulate mosquito life history and vector competence to curb virus transmission. Earlier studies have found that Wolbachia is commonly detected in Ae .  albopictus but rarely detected in Ae. aegypti . In this study, we used a two-step PCR assay to detect Wolbachia in wild-collected samples of Ae. aegypti . The PCR products were sequenced to validate amplicons and identify Wolbachia strains. A loop-mediated isothermal amplification (LAMP) assay was developed and used for detecting Wolbachia in selected mosquito specimens as well. We found Wolbachia in 85/148 (57.4%) wild Ae. aegypti specimens from various cities in New Mexico, and in 2/46 (4.3%) from St. Augustine, Florida. Wolbachia was not detected in 94 samples of Ae. aegypti from Deer Park, Harris County, Texas. Wolbachia detected in Ae. aegypti from both New Mexico and Florida was the w AlbB strain of Wolbachia pipientis . A Wolbachia -positive colony of Ae. aegypti was established from pupae collected in Las Cruces, New Mexico, in 2018. The infected females of this strain transmitted Wolbachia to their progeny when crossed with males of Rockefeller strain of Ae. aegypti , which does not carry Wolbachia . In contrast, none of the progeny of Las Cruces males mated to Rockefeller females were infected with Wolbachia ., Competing Interests: The authors declare no competing financial interests.

Although mosquito microbiota are known to influence reproduction, nutrition, disease transmission, and pesticide resistance, the relationship between host-associated microbial community composition and geographical location is poorly understood. To begin addressing this knowledge gap, we characterized microbiota associated with adult females of Culex nigripalpus mosquito vectors of Saint Louis Encephalitis and West Nile viruses sampled from three locations in Florida (Vero Beach, Palmetto Inland, and Palmetto Coast). High-throughput sequencing of PCR-amplified 16S rRNA genes demonstrated significant differences among microbial communities of mosquitoes sampled from the three locations. Mosquitoes from Vero Beach (east coast Florida) were dominated by uncultivated Asaia sp. ( Alphaproteobacteria ), whereas microbiota associated with mosquitoes collected from two mosquito populations at Palmetto (west coast Florida) sites were dominated by uncultured Spironema culicis ( Spirochaetes ), Salinisphaera hydrothermalis ( Gammaproteobacteria ), Spiroplasma ( Mollicutes ), uncultured Enterobacteriaceae , Candidatus Megaira ( Alphaproteobacteria ; Rickettsiae ), and Zymobacter ( Gammaproteobacteria ). The variation in taxonomic profiles of Cx. nigripalpus gut microbial communities, especially with respect to dominating taxa, is a potentially critical factor in understanding disease transmission and mosquito susceptibility to insecticides among different mosquito populations., Competing Interests: The authors declare that they have no competing interests.

The amount of solid waste generated in developing countries is rising over time due to economic growth, change inconsumer behavior, and lifestyles of people. But it is hard to manage and handle the increase of solid waste with existing waste management infrastructure. Thus, the management system of solid waste is very poor and has become a serious problem. The main purpose of this study is to quantify the volume of solid waste generated and investigate factors affecting generation and disposal of wastes in the study area. The result of this study indicated that total waste generated from households was about 97.092 kg/day. Furthermore, the study reveals that the solid waste generation rate of the town is 0.261 kg/person/day. About 57.5% of solid waste is properly disposed of to a landfill site, whereas the remaining 42.5% is illegally dumped at the roadsides and open fields. Implications: Nowadays, in developing countries there is a high concentration of people in urban areas, causing the generation of an enormous concentration of municipal waste in urban areas. Therefore this study's findings will be important for various policymakers and town planners. This may also serve as a benchmark for the municipal authorities of the town for whom the problem is still invisible and negligible and can push environmental protection authorities to reexamine the implementation of their policies and strategies with regard to the broader issues of human and environmental health conditions of town dwellers.

Microbiota associated with mosquito vector populations impact several traits of mosquitoes, including survival, reproduction, control, and immunity against pathogens. The influence of seasonal variations and mosquito species on mosquito gut microbiota is poorly understood. We sought to determine whether the mosquito microbiota associated with immature stages of two congeners (Culex coronator and Culex nigripalpus) differ temporally and between the two species. Using high throughput 16S rRNA gene sequence analysis, we characterized bacterial and archaeal communities found in the immature stages of the two Culex mosquito species sampled over three seasons to compare the diversity of bacteria between the two species. Beta diversity analyses of the larval microbiota sequences revealed that the two Culex species differed significantly, both temporally within each species and between the two species. Bacteria in Cx. coronator larvae were dominated by Alphaproteobacteria, mainly associated with Roseoccocus and unidentified species of Rhizobiales, and two unidentified species of Cyanobacteria. In contrast, Cx. nigripalpus was dominated by Thorsellia anophelis (Gammaproteobacteria), Clostridium, an unidentified species of Ruminococcacae (Clostridiales), and additional unidentified species associated with Erysipelotrichaceae (Erysipelotrichales), Bacteroidales, and Mollicutes. Results of our study revealed both seasonal and interspecies differences in bacterial community composition associated with the immature stages of Cx. coronator and Cx. nigripalpus vector populations in Florida. These results have important implications for our understanding of the underlying factors of variations in disease transmission among seasons, susceptibility to various pesticides, and other biotic factors, including the role of the microbiota on the spread of invasive species. In addition, our results suggest close associations of certain bacteria species with each of the two Culex species that will be further targeted for their potential in the development of microbial-based control strategies.

Aedes aegypti (L.) is the vector responsible for transmitting dengue, chikungunya, yellow fever, and Zika viruses, as well as other pathogens. Microbial larvicides based on Bacillus thuringiensis Berliner israelensis (Bti) and Saccharopolyspora spinosa Mertz and Yao, such as VectoBac 12AS and Natular 2EC, have been shown to be effective in reducing larval populations of Ae. aegypti. We examined the gene expression of two detoxification enzymes, glucosyl and glucuronosyl transferases (AaeGGT1 and AaeGGT2), through developmental stages and a time course study in response to larvicide exposure using qualitative real-time polymerase chain reaction (qPCR). AaeGGT1 and AaeGGT2 gene expressions were differentially regulated during development of the immature stages. We also found that male adults had higher expression than female adults after controlling for age effects. AaeGGT1 and AaeGGT2 gene expression were both upregulated in response to VectoBac 12AS and Natular 2EC treatments with the maximum level of expression occurring 24 h after treatment applications. This information sheds light on larvicide-induced changes in the physiology of Ae. aegypti with implications for development of mosquito control strategies., (© The Authors 2017. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

The effects of microbial biopesticides used for mosquito control on autotrophic microorganisms such as nanophytoplankton are equivocal. We examined impacts of mosquito biopesticides and mosquito larvae on primary producers in two independent experiments. In the first experiment, we examined the effects of a commonly used microbial biopesticide formulation (VectoMax ® CG) on a unicellular microalga, Selenastrum capricornatum Printz, under axenic laboratory conditions. The biopesticide treatments included two concentrations (0.008 and 0.016 g liter -1 ) of VectoMax ® CG and two controls (one untreated and another with autoclaved 0.016 g VectoMax ® CG liter -1 ) in replicated axenic experimental microcosms. Spectrophotometric analysis of chlorophyll a (proxy for algal biomass) and direct enumeration of algal cells following the treatments revealed no significant effects of the microbial biopesticide on algal population growth during the four-week study. In the second experiment, we tested the effects of different densities of Culex larvae on the population of S. capricornatum. Effects of mosquito larvae feeding on S. capricornatum were significant with a curvilinear relationship between larval density and algal abundance in the water column. Together, these studies demonstrated a lack of direct cytological/toxicological effects of Bacillus-based microbial pesticides on freshwater primary production and support the hypothesis that the reduction in algal primary production previously reported when Bti products were applied to aquatic environments was likely independent of the Bacillus-based larvicidal toxins. Instead, it was likely mediated by microbial interactions in the water column and the trophic cascade effects that resulted from the removal of larval mosquitoes. These studies suggest that mosquito larvae independent of pesticide application can influence primary production. Our method of evaluating biopesticides against small photoautotrophs can be very useful for studying the unintended effects on autotrophic microorganisms of other pesticides, including herbicides and pesticides applied to aquatic environments., (© 2017 The Society for Vector Ecology.)

Ciliate protists and rotifers are ubiquitous in aquatic habitats and can comprise a significant portion of the microbial food resources available to larval mosquitoes, often showing substantial declines in abundance in the presence of mosquito larvae. This top-down regulation of protists is reported to be strong for mosquitoes inhabiting small aquatic containers such as pitcher plants or tree holes, but the nature of these interactions with larval mosquitoes developing in other aquatic habitats is poorly understood. We examined the effects of these two microbial groups on lower trophic level microbial food resources, such as bacteria, small flagellates, and organic particles, in the water column, and on Culex larval development and adult production. In three independent laboratory experiments using two microeukaryote species (one ciliate protist and one rotifer) acquired from field larval mosquito habitats and cultured in the laboratory, we determined the effects of Culex nigripalpus larval grazing on water column microbial dynamics, while simultaneously monitoring larval growth and development. The results revealed previously unknown interactions that were different from the top-down regulation of microbial groups by mosquito larvae in other systems. Both ciliates and rotifers, singly or in combination, altered other microbial populations and inhibited mosquito growth. It is likely that these microeukaryotes, instead of serving as food resources, competed with early instar mosquito larvae for microbes such as small flagellates and bacteria in a density-dependent manner. These findings help our understanding of the basic larval biology of Culex mosquitoes, variation in mosquito production among various larval habitats, and may have implications for existing vector control strategies and for developing novel microbial-based control methods.

Pollution from nutrients in aquatic habitats has been linked to increases in disease vectors, including mosquitoes and other pestiferous insects. One possibility is that changes in mosquito microbiomes are impacted by nutrient enrichments and that these changes affect various traits, including larval development, susceptibility to larval control agents, and susceptibility of the adult mosquitoes to pathogens. We tested this hypothesis using field mesocosms supplemented with low- and high-organic-nutrient regimens and then sampled microbial communities associated with the naturally colonizing Culex nigripalpus mosquito vector. By high-throughput sequencing of 16S rRNA gene sequences, we found no significant differences in overall microbial communities associated with sampled mosquitoes, despite detecting discernible differences in environmental variables, including pH, dissolved oxygen, and nutrient amendments. Nevertheless, indicator species analysis revealed that members of the Clostridiales were significantly associated with mosquitoes that originated from high-nutrient enrichments. In contrast, members of the Burkholderiales were associated with mosquitoes from the low-nutrient enrichment. High bacterial variability associated with the life stages of the C. nigripalpus was largely unaffected by levels of nutrient enrichments that impacted larval microbial resources, including bacteria, ciliates, and flagellates in the larval environments. IMPORTANCE Mosquito microbiota provide important physiological and ecological attributes to mosquitoes, including an impact on their susceptibility to pathogens, fitness, and sensitivity to mosquito control agents. Culex nigripalpus mosquito populations transmit various pathogens, including the Saint Louis and West Nile viruses, and proliferate in nutrient-rich environments, such as in wastewater treatment wetlands. Our study examined whether increases in nutrients within larval mosquito developmental habitats impact microbial communities associated with C. nigripalpus mosquitoes. We characterized the effects of organic enrichments on microbiomes associated with C. nigripalpus mosquitoes and identified potential bacterial microbiota that will be further investigated for whether they alter mosquito life history traits and for their potential role in the development of microbial-based control strategies.

Background: The native microflora associated with mosquitoes have important roles in mosquito development and vector competence. Sequencing of bacterial V3 region from 16S rRNA genes across the developmental stages of Culex mosquitoes (early and late larval instars, pupae and adults) was used to test the hypothesis that bacteria found in the larval stage of Culex are transstadially transmitted to the adult stage, and to compare the microbiomes of field-collected versus laboratory-reared mosquitoes., Results: Beta diversity analysis revealed that bacterial community structure differed among three life stages (larvae, pupae and adults) of Culex tarsalis. Although only ~2% of the total number of bacterial OTUs were found in all stages, sequences from these OTUs accounted for nearly 82% of the total bacterial sequences recovered from all stages. Thorsellia (Gammaproteobacteria) was the most abundant bacterial taxon found across all developmental stages of field-collected Culex mosquitoes, but was rare in mosquitoes from laboratory-reared colonies. The proportion of Thorsellia sequences in the microbiomes of mosquito life stages varied ontogenetically with the greatest proportions recovered from the pupae of C. tarsalis and the lowest from newly emerged adults. The microbiome of field-collected late instar larvae was not influenced significantly by differences in the microbiota of the habitat due to habitat age or biopesticide treatments. The microbiome diversity was the greatest in the early instar larvae and the lowest in laboratory-reared mosquitoes., Conclusions: Bacterial communities in early instar C. tarsalis larvae were significantly more diverse when compared to late instar larvae, pupae and newly emerged adults. Some of the bacterial OTUs found in the early instar larvae were also found across developmental stages. Thorsellia dominated the bacterial communities in field-collected immature stages but occurred at much lower relative abundance in adults. Differences in microbiota observed in larval habitats did not influence bacterial community profiles of late instar larvae or adults. However, bacterial communities in laboratory-reared C. tarsalis larvae differed significantly from the field. Determining the role of Thorsellia in mosquitoes and its distribution across different species of mosquitoes warrants further investigation.

Schoenoplectus maritimus (alkali bulrush) has desirable attributes, such as a short growth habit (height of mature stands < 1.5 m) and annual senescence, for a potential alternative to tall (height > 3 m) emergent macrophytes in shallow constructed treatment wetlands treating ammonium-dominated wastewater. The effects of different ammonium nitrogen (NH4-N) levels on alkali bulrush growth and its ability to take up nutrients from the wastewater, as well as on mosquito production, across the range of NH4-N found in constructed wetlands of southern California are unknown. We evaluated the effects of enrichment with NH4-N on mosquito production and on the nutrient uptake and growth of alkali bulrush in two studies. Overall, significantly greater numbers (> 50%) of immature mosquitoes (mainly Culex tarsalis) were found in mesocosms enriched with NH4-N than in mesocosms receiving ambient (<0.3 mg/liter) NH4-N. High NH4-N enrichment (up to 60 mg/liter) did not adversely impact the height and stem density of S. maritimus, although a significant decrease in biomass was observed at the highest enrichment level. Nitrogen uptake by alkali bulrush increased directly with NH4-N enrichment, whereas carbon was conserved in the above-ground biomass across the enrichment gradient. Alkali bulrush is recommended for use as part of integrated mosquito management programs for moderately enriched, multipurpose, constructed treatment wetlands that improve water quality as well as provide wetland habitat for waterfowl., (© 2014 The Society for Vector Ecology.)

Microbes are important for mosquito nutrition, growth, reproduction and control. In this study, we examined bacterial communities associated with larval mosquitoes and their habitats. Specifically, we characterized bacterial communities associated with late larval instars of the western encephalitis mosquito (Culextarsalis), the submerged portions of two emergent macrophytes (California bulrush, Schoenoplectuscalifornicus and alkali bulrush, Schoenoplectusmaritimus), and the associated water columns to investigate potential differential use of resources by mosquitoes in different wetland habitats. Using next-generation sequence data from 16S rRNA gene hypervariable regions, the alpha diversity of mosquito gut microbial communities did not differ between pond mesocosms containing distinct monotypic plants. Proteobacteria, dominated by the genus Thorsellia (Enterobacteriaceae), was the most abundant phylum recovered from C. tarsalis larvae. Approximately 49% of bacterial OTUs found in larval mosquitoes were identical to OTUs recovered from the water column and submerged portions of the two bulrushes. Plant and water samples were similar to one another, both being dominated by Actinobacteria, Bacteroidetes, Cyanobacteria, Proteobacteria and Verrucomicrobia phyla. Overall, the bacterial communities within C. tarsalis larvae were conserved and did not change across sampling dates and between two distinct plant habitats. Although Thorsellia spp. dominated mosquito gut communities, overlap of mosquito gut, plant and water-column OTUs likely reveal the effects of larval feeding. Future research will investigate the role of the key indicator groups of bacteria across the different developmental stages of this mosquito species.

Mineral deficiencies in cattle, widespread in East Africa, impair optimal health and production and consequently place a great burden on the farmers' income. Therefore, detection of shortages and imbalances of specific minerals is essential. Our objective was to evaluate the mineral status of grazing cattle around the Gilgel Gibe catchment in Ethiopia and associated factors. In study I, individual animal plasma and herd faecal Ca, P, Mg, Na, K, S, Fe, Zn, Mn and Cu concentrations were determined in adult zebu cattle (Bos indicus; n=90) grazing at three altitudes around the catchment, whilst recording body condition score and sex. In study II, liver samples of adult male zebu cattle (n=53) were analysed for Cu, Zn, Fe, Se and Mo concentrations and inspected for parasitic infections. Plasma and liver analyses revealed a Cu deficiency problem in the area, since 68 and 47 % of cattle, respectively, were Cu deprived according to diagnostic criteria for Bos taurus cattle. High hepatic Mo concentrations in 17 % of cases might reflect excessive dietary Mo intake. Liver Se and plasma Na concentrations were too low in 92 and 80 % of cattle. Plasma Mn concentrations were largely below the detection limit. Plasma Cu as well as Ca concentrations were lower in the lowest altitude compared to the highest altitude group (P<0.05), whereas lean to medium cattle had lower plasma Cu concentrations (P<0.05). No differences in hepatic mineral concentrations were detected between cattle with different types of parasitic infection. In conclusion, bovine mineral deficiencies were present in the Gilgel Gibe area and were associated with grazing altitude and body condition score.