Detection of Invasive Anopheles stephensi Mosquitoes through Molecular Surveillance, Ghana.

The invasive Anopheles stephensi mosquito has rapidly expanded in range in Africa over the past decade. Consistent with World Health Organization guidelines, routine entomologic surveillance of malaria vectors in Accra, Ghana, now includes morphologic and molecular surveillance of An. stephensi mosquitoes. We report detection of An. stephensi mosquitoes in Ghana.

Biting behaviour, spatio-temporal dynamics, and the insecticide resistance status of malaria vectors in different ecological zones in Ghana.

BACKGROUND: A significant decrease in malaria morbidity and mortality has been attained using long-lasting insecticide-treated nets and indoor residual spraying. Selective pressure from these control methods influences changes in vector bionomics and behavioural pattern. There is a need to understand how insecticide resistance drives behavioural changes within vector species. This study aimed to determine the spatio-temporal dynamics and biting behaviour of malaria vectors in different ecological zones in Ghana in an era of high insecticide use for public health vector control. METHODS: Adult mosquitoes were collected during the dry and rainy seasons in 2017 and 2018 from five study sites in Ghana in different ecological zones. Indoor- and outdoor-biting mosquitoes were collected per hour from 18:00 to 06:00 h employing the human landing catch (HLC) technique. Morphological and molecular species identifications of vectors were done using identification keys and PCR respectively. Genotyping of insecticide-resistant markers was done using the TaqMan SNP genotyping probe-based assays. Detection of Plasmodium falciparum sporozoites was determined using PCR. RESULTS: A total of 50,322 mosquitoes belonging to four different genera were collected from all the study sites during the sampling seasons in 2017 and 2018. Among the Anophelines were Anopheles gambiae s.l. 93.2%, (31,055/33,334), An. funestus 2.1%, (690/33,334), An. pharoensis 4.6%, (1545/33,334), and An. rufipes 0.1% (44/33,334). Overall, 76.4%, (25,468/33,334) of Anopheles mosquitoes were collected in the rainy season and 23.6%, (7866/33,334) in the dry season. There was a significant difference (Z = 2.410; P = 0.0160) between indoor-biting (51.1%; 15,866/31,055) and outdoor-biting An. gambiae s.l. (48.9%; 15,189/31,055). The frequency of the Vgsc-1014F mutation was slightly higher in indoor-biting mosquitoes (54.9%) than outdoors (45.1%). Overall, 44 pools of samples were positive for P. falciparum CSP giving an overall sporozoite rate of 0.1%. CONCLUSION: Anopheles gambiae s.l. were more abundant indoors across all ecological zones of Ghana. The frequency of G119S was higher indoors than outdoors from all the study sites, but with higher sporozoite rates in outdoor mosquitoes in Dodowa and Kpalsogu. There is, therefore, an urgent need for a supplementary malaria control intervention to control outdoor-biting mosquitoes.

First detection of Anopheles stephensi in Ghana using molecular surveillance.

The invasive Anopheles stephensi mosquito has been rapidly expanding in range in Africa over the last decade, spreading from the Indian sub-continent to several East African countries (Djibouti, Ethiopia, Sudan, Somalia and Kenya) and now in West Africa, Nigeria. The rapid expansion of this invasive vector poses a major threat to current malaria control and elimination efforts. In line with the WHO's strategy to stop the spread of this invasive species by enhancing surveillance and control measures in Africa, we incorporated morphological and molecular surveillance of An. stephensi into routine entomological surveillance of malaria vectors in the city of Accra, Ghana. Here, we report on the first detection of An. stephensi in Ghana. An. stephensi mosquitoes were confirmed using PCR and sequencing of the ITS2 regions. These findings highlight the urgent need for increased surveillance and response strategies to mitigate the spread of An. stephensi in Ghana.

Transcriptome profiling of two rice genotypes under mild field drought stress during grain-filling stage.

Drought is one of the most critical abiotic stresses that threaten crop production worldwide. This stress affects the rice crop in all stages of rice development; however, the occurrence during reproductive and grain-filling stages has the most impact on grain yield. Although many global transcriptomic studies have been performed during the reproductive stage in rice, very limited information is available for the grain-filling stage. Hence, we intend to investigate how the rice plant responds to drought stress during the grain-filling stage and how the responses change over time under field conditions. Two rice genotypes were selected for RNA-seq analysis: '4610', previously reported as a moderately tolerant breeding line, and Rondo, an elite indica rice cultivar susceptible to drought conditions. Additionally, 10 agronomic traits were evaluated under normal irrigated and drought conditions. Leaf tissues were collected during grain-filling stages at two time points, 14 and 21 days after the drought treatment, from both the drought field and normal irrigated field conditions. Based on agronomic performances, '4610' was less negatively affected than Rondo under mild drought conditions, and expression profiling largely aligned with the phenotypic data. The transcriptomic data indicated that, in general, '4610' had much earlier responses than its counterpart in mitigating the impact of drought stress. Several key genes and gene families related to drought stress or stress-related conditions were found differentially expressed in this study, including transcription factors, drought tolerance genes and reactive oxygen species scavengers. Furthermore, this study identified novel differentially expressed genes (DEGs) without function annotations that may play roles in drought tolerance-related functions. Some of the important DEGs detected in this study can be targeted for future research.

Novel Wolbachia strains in Anopheles malaria vectors from Sub-Saharan Africa.

Background:  Wolbachia, a common insect endosymbiotic bacterium that can influence pathogen transmission and manipulate host reproduction, has historically been considered absent from the  Anopheles (An.) genera, but has recently been found in  An. gambiae s.l. populations in West Africa.  As there are numerous  Anopheles species that have the capacity to transmit malaria, we analysed a range of species across five malaria endemic countries to determine  Wolbachia prevalence rates, characterise novel  Wolbachia strains and determine any correlation between the presence of  Plasmodium,  Wolbachia and the competing bacterium  Asaia. Methods:  Anopheles adult mosquitoes were collected from five malaria-endemic countries: Guinea, Democratic Republic of the Congo (DRC), Ghana, Uganda and Madagascar, between 2013 and 2017.  Molecular analysis was undertaken using quantitative PCR, Sanger sequencing,  Wolbachia multilocus sequence typing (MLST) and high-throughput amplicon sequencing of the bacterial  16S rRNA gene.  Results: Novel  Wolbachia strains were discovered in five species:  An. coluzzii,  An. gambiae s.s.,  An. arabiensis,  An. moucheti and  An. species A, increasing the number of  Anopheles species known to be naturally infected. Variable prevalence rates in different locations were observed and novel strains were phylogenetically diverse, clustering with  Wolbachia supergroup B strains.  We also provide evidence for resident strain variants within  An. species A.  Wolbachia is the dominant member of the microbiome in  An. moucheti and  An. species A but present at lower densities in  An. coluzzii.  Interestingly, no evidence of  Wolbachia/Asaia co-infections was seen and  Asaia infection densities were shown to be variable and location dependent.  Conclusions: The important discovery of novel  Wolbachia strains in  Anopheles provides greater insight into the prevalence of resident  Wolbachia strains in diverse malaria vectors.  Novel  Wolbachia strains (particularly high-density strains) are ideal candidate strains for transinfection to create stable infections in other  Anopheles mosquito species, which could be used for population replacement or suppression control strategies.

An unusual cause of a sellar mass.

A 57-year-old man presented with features suggestive of a non-functioning pituitary tumour. He underwent trans-sphenoidal resection of the tumour which was found to be a chordoma and not a pituitary adenoma. This case demonstrates that a chordoma may directly involve the sellar region and mimic a pituitary adenoma. We also discuss the aetiology, areas of involvement, management and prognosis of this unusual tumour.

Instrumentation enabling study of plant physiological response to elevated night temperature.

BACKGROUND: Global climate warming can affect functioning of crops and plants in the natural environment. In order to study the effects of global warming, a method for applying a controlled heating treatment to plant canopies in the open field or in the greenhouse is needed that can accept either square wave application of elevated temperature or a complex prescribed diurnal or seasonal temperature regime. The current options are limited in their accuracy, precision, reliability, mobility or cost and scalability. RESULTS: The described system uses overhead infrared heaters that are relatively inexpensive and are accurate and precise in rapidly controlling the temperature. Remote computer-based data acquisition and control via the internet provides the ability to use complex temperature regimes and real-time monitoring. Due to its easy mobility, the heating system can randomly be allotted in the open field or in the greenhouse within the experimental setup. The apparatus has been successfully applied to study the response of rice to high night temperatures. Air temperatures were maintained within the set points +/- 0.5 degrees C. The incorporation of the combination of air-situated thermocouples, autotuned proportional integrative derivative temperature controllers and phase angled fired silicon controlled rectifier power controllers provides very fast proportional heating action (i.e. 9 ms time base), which avoids prolonged or intense heating of the plant material. CONCLUSION: The described infrared heating system meets the utilitarian requirements of a heating system for plant physiology studies in that the elevated temperature can be accurately, precisely, and reliably controlled with minimal perturbation of other environmental factors.

Leaf and canopy photosynthetic characteristics of cotton (Gossypium hirsutum) under elevated CO2 concentration and UV-B radiation.

Increases in both atmospheric CO2 concentration ([CO2]) and ultraviolet-B (UV-B) radiation on the Earth's surface are features of current climate change patterns. An experiment was conducted in sunlit, controlled environment chambers known as Soil-Plant-Atmosphere-Research (SPAR) units to determine interactive effects of elevated [CO2] and UV-B radiation on leaf and canopy photosynthetic characteristics of cotton. Six treatments were comprised of two CO2 levels of 360 (ambient) and 720 (elevated) microL L(-1) and three levels of 0 (control), 8, and 16 kJ m(-2) d(-1) biologically effective UV-B radiation. Treatments were imposed for 66 days from crop emergence through three weeks after the first flower stage. Plants grown in elevated [CO2] had significantly greater leaf area, higher leaf and canopy net photosynthetic rates (PN), lower dark respiration rate (Rd), and lower light compensation point (LCP) than plants grown in ambient [CO2]. There was no difference in CO2 compensation point (gamma), maximum rate of Rubisco activity (Vcmax), or light-saturated rate of electron transport (Jmax) between ambient and elevated CO2 treatments. When plants were grown in 8 kJ m(-2) d(-1) UV-B radiation, most of the measured photosynthetic parameters did not differ from control plants. High UV-B (16 kJ) radiation, however, caused 47-50% smaller leaf area, 38-44% lower leaf PN, 72-74% lower Vcmax, and 61-66% lower Jmax compared to the control. There were no interactive effects of [CO2] and UV-B radiation on most of the photosynthetic parameters measured. From the results, it is concluded that decreased canopy photosynthesis due to enhanced UV-B radiation in cotton is associated with both smaller leaf area and lower leaf PN, and loss of Rubisco activity and electron transport are two major factors in UV-B inhibition of leaf PN.