Biology subjects

Makhulu, E. E.

Publications and source records attributed to Makhulu, E. E..

6 recordsLinked to original sources

bioRxiv2025

Changes in the Anopheles arabiensis transcriptome and gut microbiota profiles associated with Microsporidia MB

Microsporidia MB is an endosymbiotic microbe found in Anopheles mosquito populations. This symbiont can block Plasmodium transmission in mosquitoes, and it can spread through mosquito populations and be sustained over generations by vertical and horizontal transmission. These characteristics make Microsporidia MB a potential candidate for symbiont-based malaria vector control. However, the mechanistic basis of interactions between Microsporidia MB and Anopheles arabiensis is poorly characterized. We investigated how the presence of Microsporidia MB affects transcriptomics profiles and the gut microbiota composition and diversity in non-blood-fed and blood-fed (24, 48 and 72 hours post blood meal) An. arabiensis mosquitoes. We observed that mosquito infection with Microsporidia MB upregulated farnesoic acid O-methyltransferase, a gene linked to juvenile hormone biosynthesis in the non-blood-fed mosquitoes. In addition, blood feeding in Microsporidia MB positive mosquitoes was associated with an activation of immune-related genes 24 hours after a blood meal, where several genes including the lipopolysaccharide tumor necrosis factor gene were upregulated. Interestingly, we also observed that Microsporidia MB positivity was associated with a microbiota shift to favor the proliferation of microbes including Pseudomonas and Serratia 24 hours post blood meal. There were indications of immune system activation in Microsporidia MB positive mosquitoes up to 48 hours after a blood meal where factors such as the peptidoglycan recognition SC2-like and lysozyme c-1 were upregulated. Notably, an upregulated immune system at this time point was associated with downregulation of genes associated with metabolism and the restoration of Serratia, Pseudomonas and other key microbes to relative abundances similar to those recorded in non-blood-fed mosquitoes. At the 72-hour time point, Microsporidia MB positive mosquitoes exhibited a downregulation of genes associated with immunity, including cecropins and defensins, while metabolic processes were predominantly upregulated. Our results provide insights into the effect of Microsporidia MB infection on the An. arabiensis gene expression and gut microbiota profiles. This work will contribute to mechanistic insights into symbiont-mediated malaria transmission blocking.

systems biology↗

bioRxiv2024

Temperature modulates the dissemination potential of Microsporidia MB, a malaria-blocking endosymbiont of Anopheles mosquitoes

The endosymbiont Microsporidia MB is a promising malaria control strategy that inhibits the development of Plasmodium naturally in Anopheles mosquitoes. To be successful, it would be necessary to significantly increase the prevalence of Microsporidia MB in populations of malaria mosquitoes to decrease the malaria transmission potential of the mosquito population. However, very little is known about the role of temperature in driving the prevalence of Microsporidia MB infections in mosquito populations. By rearing mosquito larvae under four air temperature regimes (22{degrees}C, 27{degrees}C, 32{degrees}C and 37{degrees}C), we show that warm temperatures favour the growth of Microsporidia MB infected larvae. In addition, Microsporidia MB infected larvae developed faster compared to the uninfected offspring of the same mothers. Starting with 10 Microsporidia MB infected mothers, our population growth model showed that, at 32{degrees}C, it would take 15-35 days to reach a population of 1000 Microsporidia MB infected mothers; this represents a dissemination potential of 4.7, 1.3 and 1.7 times higher compared to 22{degrees}C, 27{degrees}C and 37{degrees}C, respectively. Despite a relatively high mosquito mortality rate (20% more compared to 27{degrees}C), 32{degrees}C was estimated the best temperature for rearing Microsporidia MB infected larvae due to the shorter development time and high infection rate. This study gives insight into the favourable conditions for Microsporidia MB mass rearing and potential release strategies in malarious regions. ImportanceMalaria parasites transmitted by Anopheles mosquitoes cause a life-threatening disease, imposing a massive toll on human health and economic sustainability in sub-Saharan Africa. Relying only on insecticide- and drug-based control products whose efficacy has been eroded by resistance to control malaria is not sufficient anymore. New innovative approaches are urgently needed and Microsporidia MB, a naturally occurring symbiont across Africa is capable of inhibiting Plasmodium transmission in Anopheles gambiae s.l.. Its success in adverting a rebound of malaria cases will depend on the infection dynamic of the symbiont over time and space. Through experimental studies on field derived mosquitoes and mathematical modelling, we demonstrate that Microsporidia MB dissemination potential increase with temperature within a viable range for Anopheles mosquitoes, due to trade-offs between mosquito development and survival and the symbiont growth. Future studies should now investigate how fluctuating temperatures modulate the Plasmodium transmission blocking performance in nature.

microbiology↗

bioRxiv2024

The Plasmodium transmission-blocking symbiont, Microsporidia MB, is vertically transmitted through Anopheles arabiensis germline stem cells

Microsporidia MB is a promising candidate for developing a symbiont-based strategy for malaria control because it disrupts the capacity of An. arabiensis to transmit the Plasmodium parasite. The symbiont is predominantly localized in the reproductive organs and is transmitted vertically from mother to offspring and horizontally (sexually) during mating. Due to the contribution of both transmission routes, Microsporidia MB has the potential to spread through target vector populations and become established at high prevalence. Stable and efficient vertical transmission of Microsporidia MB is important for its sustainable use for malaria control, however, the vertical transmission efficiency of Microsporidia MB can vary. In this study, we investigate the mechanistic basis of Microsporidia MB vertical transmission in An. arabiensis. We show that vertical transmission occurs through the acquisition of Microsporidia MB by Anopheles cystocyte progenitors following the division of germline stem cells. We also show that Microsporidia MB replicates to increase infection intensity in the oocyte of developing eggs when mosquitoes are given a blood meal suggesting that symbiont proliferation in the ovary is coordinated with egg development. The rate of Microsporidia MB transmission to developing eggs is on average higher than the recorded (mother to adult offspring) vertical transmission rate. This likely indicates that a significant proportion of An. arabiensis offspring lose their Microsporidia MB symbionts during development. The stability of germline stem cell infections, coordination of symbiont proliferation, and very high rate of transmission from germline stem cells to developing eggs indicate that Microsporidia MB has a highly specialized vertical transmission strategy in An. arabiensis, which may explain host specificity. Author SummaryMosquito vectors of diseases are associated with a broad range of microbes. Some of the microbes significantly affect vector biology including pathogen transmission efficiency. Anopheles mosquitoes, which transmit the malaria parasite, Plasmodium falciparum, harbor a native microbe known as Microsporidia MB. This microbe interferes with the formation of transmissible stages of the parasite that are transferred to humans by female mosquitoes when taking a blood meal. This phenotype can be exploited to develop a novel strategy for controlling malaria similar to the control of dengue fever using Aedes mosquitoes carrying Wolbachia bacteria. Mother-to-offspring transmission of protective microbes is important in sustainable application of microbe-based technologies to control vector-borne diseases because it ensures maintenance of the microbe in target vector populations across many generations. Here, we investigated stability of Microsporidia MB infections and efficiency of mother-to-offspring transmission during early stages of egg formation and development. We found that this microbe has a specialized transmission mechanism that involves infecting the germline cells that are important in egg production. We also demonstrated a very high transmission rate (97%) of the Microsporidia MB from infected germline cells into daughter cells during cell division. As the germline daughter cells developed into eggs, Microsporidia MB established itself in the egg yolk through active replication which only occured after the female mosquitoes had a blood meal. Our study gives insights into an efficient mother-to-offspring transmission route of Microsporidia MB that can be utilized sustainably in microbe-based intervention to control malaria.

ecology↗

bioRxiv2023

Localisation and tissue tropism of the symbiont Microsporidia MB in the germ line and somatic tissues of Anopheles arabiensis

The Anopheles symbiont, Microsporidia MB, is maternally inherited and has a strong malaria transmission-blocking phenotype in Anopheles arabiensis. Microsporidia MB is also vertically transmitted, sexually transmitted and avirulent. These characteristics are expected to promote its spread through mosquito populations, enhancing the potential of Microsporidia MB as a candidate for the development of a symbiont-mediated malaria transmission blocking strategy. We found that the patterns of Microsporidia MB localisation over the development of An. arabiensis indicate accumulation in tissues linked to its transmission, specifically the male and female gonadal tissues. Transovarial vertical transmission of Microsporidia MB occurs in the female An. arabiensis ovary when Microsporidia MB becomes localised to the cytoplasm of the developing oocyte. In male An. arabiensis, Microsporidia MB is localised in the testis and vas deferens. Notably, a high intensity of Microsporidia MB can also be observed in the An. arabiensis adult but not larval gut. The levels of Microsporidia MB found in the female ovary are linked to the progression of oogenesis, increasing after blood feeding initiates the development of eggs. The levels of Microsporiodia MB in the male and female gonadal and gut tissue do not increase as mosquitoes age. Altogether, the high specificity of Microsporidia MB tissue localisation patterns and changes in infection prevalence and intensity suggest adaptation to maximise transmission and avirulence in Anopheles arabiensis. ImportanceMicrosporidia MB is a symbiont with strong malaria transmission-blocking phenotype in Anopheles arabiensis. It spreads in mosquito populations through mother-to-offspring and sexual transmission. The ability of Microsporidia MB to block Plasmodium transmission together with its ability to spread within Anopheles populations and its avirulence to the host makes it a very attractive candidate for developing a key strategy to stop malaria transmissions. Here, we report the basis of Microsporidia MB transmission. We find that Microsporidia MB accumulates in Anopheles arabiensis tissues linked to its sexual and vertical transmission. Its prevalence and intensity in the tissues over the mosquito life cycle suggest adaptation to maximise transmission and avirulence in Anopheles arabiensis. These findings provide the foundation for understanding the factors that affect Microsporidia MB transmission efficiency. This will contribute to the establishment of strategies to maximize Microsporidia MB transmission for Anopheles mosquito population replacement and malaria transmission blocking.

microbiology↗

bioRxiv2020

Tsetse blood-meal sources, endosymbionts, and trypanosome infections provide insight into African trypanosomiasis transmission in the Maasai Mara National Reserve, a wildlife-human-livestock interface

BackgroundAfrican trypanosomiasis (AT) is a neglected disease of both humans and animals caused by Trypanosoma parasites, which are transmitted by obligate hematophagous tsetse flies (Glossina spp.). Understanding of AT transmission is hampered by limited knowledge on interactions of tsetse flies with their vertebrate hosts and the influence of endosymbionts on vector competence, especially in wildlife-human-livestock interfaces. We identified the tsetse species, their blood-meal sources, and the correlation between endosymbiont and trypanosome infection status in the trypanosome-endemic Maasai Mara National Reserve (MMNR) of Kenya. Methodology/Principal FindingsAmong 1167 tsetse flies (1136 Glossina pallidipes, 31 Glossina swynnertoni) collected from 10 sampling sites, 28 (2.4%) were positive by PCR for trypanosomes, majority (17/28) being Trypanosoma vivax. Blood-meal analyses based on high-resolution melting analysis of mitochondrial cytochrome c oxidase 1 and cytochrome b gene PCR products (n = 345) identified humans as the most common vertebrate host (37%), followed by hippopotamus (29.1%), African buffalo (26.3%), elephant (3.39%), and giraffe (0.84%). Trypanosome-infected flies had fed on hippopotamus and buffalo. Additionally, PCR analysis revealed that tsetse flies were more likely to be infected with trypanosomes if they were infected with the Sodalis glossinidius endosymbiont (P = 0.0022 Fishers exact test). Conclusions/SignificanceDiverse species of wildlife hosts may contribute to the maintenance of tsetse populations and/or persistent circulation of African trypanosomes in the MMNR. Although the African buffalo is known to be a key reservoir of AT, the higher proportion of hippopotamus blood-meals in trypanosomes-infected flies identified here indicates that other wildlife species may also be important to transmission cycles. No trypanosomes associated with human disease were identified, but the high proportion of human blood-meals identified are indicative of human African trypanosomiasis transmission risk. Furthermore, this work provides data showing that Sodalis endosymbionts can is associated with increased trypanosome infection rates in endemic ecologies. Author summaryHuman and animal African trypanosomiasis are neglected tropical diseases with potential to spread to new areas. Wild animals are important reservoirs for African trypanosomes and crucial in the emergence and re-emergence of AT. Vertebrate host-vector-parasite interactions are integral to trypanosome transmission. We investigated the vertebrate blood-meals and trypanosomes-endosymbionts co-infections in tsetse flies, which have been associated with reservoirs and vector competence, respectively, on AT transmission in Kenyas Maasai Mara National Reserve. We identified tsetse fly diversity, trypanosome and endosymbiont infection status, and vertebrate blood-meal hosts to infer potential transmission dynamics. We found that Glossina pallidipes was the major tsetse fly vector and that Trypanosoma vivax was the main trypanosome species circulating in the region. Humans, hippopotamus, and buffalo were the most frequented for blood-meals. Buffalo and hippopotamus blood-meals were identified in trypanosome infected flies. Feeding of the flies on both humans and wildlife may potentiate the risk of the human trypanosomiasis in this ecology. Additionally, we found that the endosymbiont Sodalis glossinidius is associated with higher trypanosome infection rates in wild tsetse flies. These findings emphasize the importance of understanding the interaction of tsetse flies with vertebrate blood-meal sources and their endosymbionts in the transmission and control of AT.

molecular biology↗

bioRxiv2020

Pathogens, endosymbionts, and blood-meal sources of host-seeking ticks in the fast-changing Maasai Mara wildlife ecosystem

BackgroundThe role of questing ticks in the epidemiology of tick-borne diseases in Kenyas Maasai Mara National Reserve (MMNR), an ecosystem with intensified human-wildlife-livestock interactions, remains poorly understood. Therefore, we carried out a survey of the diversity of questing ticks, their blood-meal hosts, and tick-borne pathogens to understand potential effects to human and livestock health. MethodsQuesting ticks were collected by flagging and hand picks from vegetation in 25 localities and identified based on morphologic and molecular criteria. We used PCR with high-resolution melting (HRM) analysis, and sequencing to identify Anaplasma, Babesia, Coxiella, Ehrlichia, Rickettsia, and Theileria pathogen diversities and blood meals in 231 tick pools. ResultsA total of 1,465 host-seeking ticks were collected, including Rhipicephalus appendiculatus (n = 1,125), Rhipicephalus pulchellus (n = 6), Rhipicephalus evertsi (n = 5), Amblyomma cf. gemma (n = 178), Amblyomma gemma (n = 145), Amblyomma variegatum (n = 4), Amblyomma sp. (n = 1), and Haemaphysalis leachi (n = 1). Remnant blood-meals from humans, wildebeest, and African buffalo were detected in Rh. appendiculatus, goat in Rh. evertsi, sheep in Am. gemma, and cattle in Am. variegatum. Rickettsia africae was detected in Am. gemma (1/25 pools) that had blood-meal remnant from sheep and Am. variegatum (4/25 pools) that had fed on cattle. Rickettsia spp. were found in Am. gemma (4/25 pools) and Rh. evertsi (1/4 pools). Anaplasma ovis was detected in Rh. appendiculatus (1/172 pools) and Rh. evertsi (1/4 pools), while Anaplasma bovis was detected in Rh. appendiculatus (1/172 pools). Theileria parva was detected in Rh. appendiculatus (27/172 pools). Babesia, Ehrlichia and Coxiella pathogens were not found in any ticks. Unexpectedly, diverse Coxiella sp. endosymbionts were detected in all tick genera (174/231 pools). ConclusionsThe data shows that ticks from the rapidly-changing MMNR are infected with zoonotic R.africae and unclassified Rickettsia spp, demonstrating the persistent risk of African tick-bite fever and other and Spotted Fever Group rickettsioses to local dwellers and visitors to the Maasai Mara ecosystem. Protozoan pathogens that may pose risk to livestock production were also identified. We also highlight possible existence of morphotypic variants of Amblyomma species, based on the observation of Ambyomma cf. gemma, which may be potential human parasites or emergent disease vectors. Our findings also demonstrate that questing ticks in this ecosystem have dynamic vertebrate blood sources including humans, wildlife and domestic animals, which may amplify transmission of tickborne zoonoses and livestock diseases. Further studies are needed to determine the role of Coxiella endosymbionts in tick physiology and vector competence.

molecular biology↗