Wolbachia endosymbionts in two Anopheles species indicates independent acquisitions and lack of prophage elements

Wolbachia is a genus of obligate bacterial endosymbionts that infect a diverse range of arthropod species as well as filarial nematodes, with its single described species, Wolbachia pipientis, divided into several 'supergroups' based on multilocus sequence typing. Wolbachia strains in mosquitoes have been shown to inhibit the transmission of human pathogens including Plasmodium malaria parasites and arboviruses. Despite their large host range, Wolbachia strains within the major malaria vectors of the Anopheles (A.) gambiae and A. funestus complexes appear at low density based solely on PCR-based methods. Questions have been raised as to whether this represents a true endosymbiotic relationship. However, recent definitive evidence for two distinct, high-density strains of supergroup B Wolbachia within A. demeilloni and A. moucheti has opened exciting possibilities to explore naturally occurring Wolbachia endosymbionts in Anopheles for biocontrol strategies to block Plasmodium transmission. Here we utilise genomic analyses to demonstrate that both Wolbachia strains have retained all key metabolic and transport pathways despite their smaller genome size. We further confirm the presence of cytoplasmic incompatibility factor genes, despite noticeably few prophage regions. Additionally, phylogenetic analysis indicates that these Wolbachia strains may have been introduced into these two Anopheles species via horizontal transmission events, and unlikely to be by ancestral acquisition and subsequent loss events in the Anopheles gambiae species complex. These are the first Wolbachia genomes that enable us to study the relationship between natural strains Plasmodium malaria parasites and their Anopheline hosts.

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Community Knowledge, Perceptions, And Practices Regarding Malaria and Its Control In Jabi Tehnan District, Amhara Region, Northwest Ethiopia

10.21203/rs.3.rs-489446/v1 ◽

2021 ◽

Author(s):

Abebe Asale ◽

Zewdu Abro ◽

Bayu Enchalew ◽

Alayu Tesager ◽

Aklilu Belay ◽

...

Keyword(s):

Vector Control ◽

Malaria Control ◽

Community Education ◽

Indoor Residual Spraying ◽

Community Empowerment ◽

Malaria Episode ◽

Malaria Vectors ◽

Bed Nets ◽

Health Seeking ◽

Malaria Parasites

Abstract Background: Key program components of malaria control in Ethiopia include community empowerment and mobilization, vector control using long-lasting insecticidal nets (LLINs), indoor residual spraying (IRS), prompt diagnosis and treatment, and disease surveillance. However, the effectiveness of these interventions is often undermined by various challenges, including insecticide and drug resistance, the plasticity of malaria vectors feeding and biting behavior, and certain household factors that lead to misuse and poor utilization of LLINs. The primary objective of this study was to document households’ perceptions towards malaria and assess the prevalence of the disease and the constraints related to the ongoing interventions in Ethiopia (LLINs, IRS, community mobilization house screening). Method: The study was conducted in Jabi Tehnan district, Northwestern Ethiopi,a from November 2019 to March 2020. A total of 3,010 households distributed over 38 kebeles (villages) were randomly selected for socio-economic and demographic survey. Focus group discussions (FGDs) were conducted in 11 different health clusters taking into account agro-ecological differences. A total of 1,256 children under 10 years of age were screened for malaria parasites using microscopy in order to determine malaria prevalence. Furthermore, five-year malaria trend analysis was undertaken based on data obtained from the district health office to understand the disease dynamics.Result: Malaria knowledge in the area was high as all FGD participants correctly identified mosquito bites during the night as sources of malaria transmission. Delayed health seeking behavior remains a key behavioral challenge in malaria control as it took patients on average 4 days before reporting the case at the nearby health facility. On average households lost 2.53 working days per person-per malaria episode and theey spent US$ 18 per person perepisode. Out of the 1,256 randomly selected under 10 children tested for malaria parasites, 11 (0.89%) were found to be positive. Malaria disproportionately affected the adult segment of the population more, 50% of the total cases reported from households whose age was 15 and beyond. The second most affected group was the age group between 5 and 14 years followed by children under 10, with 31% and 14% burden,respectively.Conclusion: Despite the achievement of universal coverage in terms of LLINs access, utilization of vector control interventions in the area remained low.Using bed nets for unintended purposes remained a major challenge. Therefore, continued community education and communication work should be prioritised in the study area to bring about the desired behavioral changes.

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Experimental Infection of Sand Flies by Massilia Virus and Viral Transmission by Co-Feeding on Sugar Meal

Viruses ◽

10.3390/v11040332 ◽

2019 ◽

Vol 11 (4) ◽

pp. 332 ◽

Cited By ~ 2

Author(s):

Magdalena Jancarova ◽

Laurence Bichaud ◽

Jana Hlavacova ◽

Stephane Priet ◽

Nazli Ayhan ◽

...

Keyword(s):

Infection Rate ◽

Sexual Transmission ◽

Horizontal Transmission ◽

Experimental Studies ◽

Convincing Evidence ◽

Sand Fly ◽

Sand Flies ◽

Human Pathogens ◽

Larval Food ◽

Efficient Route

Background: Massilia virus (MASV) is a phlebovirus isolated from Phlebotomus perniciosus in various regions of southwestern Europe. It is closely related to human pathogens such as Toscana virus and sandfly fever Naples virus. The natural cycle of phleboviruses is poorly understood. Indeed, experimental studies demonstrate that transovarial and sexual transmission are not efficient enough for the maintenance of the virus in nature and to date there is no convincing evidence that a species of vertebrates is the reservoir of the virus. Here, we studied various transmission routes of MASV taking advantage of experimental colonies representing different species of sand flies. Methodology/Principal findings: In P. perniciosus, four sources of infection were compared: (i) Virus-seeded larval food to the first instar larvae (L1), or (ii) to the fourth instar larvae (L4), (iii) virus-seeded blood meal to adult females, and (iv) virus-seeded sugar meal to adults of both sexes. From 875 adults emerged from infected L1 and L4, only three were positive. In females infected by bloodmeal the infection rate was high before defecation, then it decreased drastically; MASV RNA was detected in only 5 out of 27 post-defecation. Surprisingly, the most efficient route of infection was observed after intake of virus-seeded sugar meal: 72% of females (79/110) and 52% of males (51/99) were found to be MASV RNA-positive. In addition, MASV-infected sandflies regurgitated virus particules into the sugar drop and MASV RNA was detectable in this drop for at least 24 h after regurgitation. MASV RNA was detected in about one third of the P. perniciosus exposed to this sugar drop contaminated by regurgitation. Sugar meal infection was also tested with six other species of sand flies. In males, there were no significant differences in infection rates when compared to P. perniciosus. In females, most species tested showed high infection rate at the beginning but then significant gradual decrease in infection rate during the experiment. Conclusions/Significance: We present the first description of arboviral infection of a dipteran vector using sugar meal. In all seven sand fly species tested, MASV was detected for two weeks post-infection. Our results showed that MASV can be transmitted between P. perniciosus either through co-feeding or via an infected sugar source such as plant sap. These newly described routes of horizontal transmission may play an important role in the circulation of phleboviruses in nature.

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Tissue Tropisms and Transstadial Transmission of a Rickettsia Endosymbiont in the Highland Midge, Culicoides impunctatus (Diptera: Ceratopogonidae)

Applied and Environmental Microbiology ◽

10.1128/aem.01492-20 ◽

2020 ◽

Vol 86 (20) ◽

Author(s):

Jack Pilgrim ◽

Stefanos Siozios ◽

Matthew Baylis ◽

Gregory D. D. Hurst

Keyword(s):

Vertical Transmission ◽

Fat Body ◽

Germ Line ◽

Horizontal Transmission ◽

Follicle Cells ◽

Biting Midges ◽

Diverse Range ◽

Suspensory Ligament ◽

Transstadial Transmission ◽

Tissue Tropisms

ABSTRACT Rickettsia is a genus of intracellular bacteria which can manipulate host reproduction and alter sensitivity to natural enemy attack in a diverse range of arthropods. The maintenance of Rickettsia endosymbionts in insect populations can be achieved through both vertical and horizontal transmission routes. For example, the presence of the symbiont in the follicle cells and salivary glands of Bemisia whiteflies allows Belli group Rickettsia transmission via the germ line and plants, respectively. However, the transmission routes of other Rickettsia bacteria, such as those in the Torix group of the genus, remain underexplored. Through fluorescence in situ hybridization (FISH) and transmission electron microscopy (TEM) screening, this study describes the pattern of Torix Rickettsia tissue tropisms in the highland midge, Culicoides impunctatus (Diptera: Ceratopogonidae). Of note is the high intensity of infection of the ovarian suspensory ligament, suggestive of a novel germ line targeting strategy. Additionally, localization of the symbiont in tissues of several developmental stages suggests transstadial transmission is a major route for ensuring maintenance of Rickettsia within C. impunctatus populations. Aside from providing insights into transmission strategies, the presence of Rickettsia bacteria in the fat body of larvae indicates potential host fitness and vector capacity impacts to be investigated in the future. IMPORTANCE Microbial symbionts of disease vectors have garnered recent attention due to their ability to alter vectorial capacity. Their consideration as a means of arbovirus control depends on symbiont vertical transmission, which leads to spread of the bacteria through a population. Previous work has identified a Rickettsia symbiont present in several species of biting midges (Culicoides spp.), which transmit bluetongue and Schmallenberg arboviruses. However, symbiont transmission strategies and host effects remain underexplored. In this study, we describe the presence of Rickettsia in the ovarian suspensory ligament of Culicoides impunctatus. Infection of this organ suggests the connective tissue surrounding developing eggs is important for ensuring vertical transmission of the symbiont in midges and possibly other insects. Additionally, our results indicate Rickettsia localization in the fat body of Culicoides impunctatus. As the arboviruses spread by midges often replicate in the fat body, this location implies possible symbiont-virus interactions to be further investigated.

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Symbiosis Comes of Age at the 10th Biennial Meeting of Wolbachia Researchers

Applied and Environmental Microbiology ◽

10.1128/aem.03071-18 ◽

2019 ◽

Vol 85 (8) ◽

Cited By ~ 4

Author(s):

Irene L. G. Newton ◽

Barton E. Slatko

Keyword(s):

Cell Biology ◽

Rna Virus ◽

Host Cells ◽

Human Pathogens ◽

Content Type ◽

New Science ◽

Successful Infection ◽

Filarial Nematodes ◽

Insect Populations ◽

Basic Biology

ABSTRACT Wolbachia pipientis is an alphaproteobacterial obligate intracellular microbe and arguably the most successful infection on our planet, colonizing 40% to 60% of insect species. Wolbachia spp. are also present in most, but not all, filarial nematodes, where they are obligate mutualists and are the targets for antifilarial drug discovery. Although Wolbachia spp. are related to important human pathogens, they do not infect mammals but instead are well known for their reproductive manipulations of insect populations, inducing the following phenotypes: male killing, feminization, parthenogenesis induction, and cytoplasmic incompatibility (CI). The most common of these, CI, results in a sperm-egg incompatibility and increases the relative fecundity of infected females in a population. In the last decade, Wolbachia spp. have also been shown to provide a benefit to insects, where the infection can inhibit RNA virus replication within the host. Wolbachia spp. cannot be cultivated outside host cells, and no genetic tools are available in the symbiont, limiting approaches available for their study. This means that many questions fundamental to our understanding of Wolbachia basic biology remained unknown for decades. The 10th biennial international Wolbachia conference, Wolbachia Evolution, Ecology, Genomics and Cell Biology: A Chronicle of the Most Ubiquitous Symbiont, was held on 17 to 22 June 2018 in Salem, MA. In this review, we highlight the new science presented at the meeting, link it to prior efforts to answer these questions across the Wolbachia genus, and present the importance of these findings to the field of symbiosis. The topics covered in this review are based on the presentations at the conference.

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Interleukin-5 Is Essential for Vaccine-Mediated Immunity but Not Innate Resistance to a Filarial Parasite

Infection and Immunity ◽

10.1128/iai.68.5.2513-2517.2000 ◽

2000 ◽

Vol 68 (5) ◽

pp. 2513-2517 ◽

Cited By ~ 47

Author(s):

Laetitia Le Goff ◽

P'ng Loke ◽

H. Fahimeda Ali ◽

David W. Taylor ◽

Judith E. Allen

Keyword(s):

Critical Role ◽

Human Pathogens ◽

Filarial Infection ◽

Innate Resistance ◽

Interleukin 5 ◽

Filarial Nematodes ◽

Third Stage ◽

Parasite Survival ◽

Patent Infection

ABSTRACT The study of protective immune mechanisms effective against filarial nematodes has been hampered by the inability of these important human pathogens to infect laboratory mice. Recently,Litomosoides sigmodontis, a natural parasite of rats, has been developed as a valuable model for the study of filarial infection. BALB/c mice are fully susceptible to infection with L. sigmodontis third-stage larvae and develop patent infection. In contrast, mice on the C57BL background are resistant, and parasites undergo only a single molt and do not mature to adulthood. We used interleukin-5 (IL-5)-deficient mice on the C57BL/6 background to address the role of IL-5 and eosinophils in the innate resistance of C57BL/6 mice. We found no differences in parasite survival between IL-5-deficient and C57BL/6 mice. However, when these mice were used for the analysis of vaccine-mediated immunity, a critical role for IL-5 was elucidated. Mice genetically deficient in IL-5 were unable to generate a protective immune response when vaccinated with irradiated larvae, whereas C57BL/6 mice were fully protected from challenge infection. These studies help to clarify the highly controversial role of eosinophils in filarial infection.

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The Tissue Tropisms and Transstadial Transmission of a Rickettsia Endosymbiont in the Highland Midge, Culicoides impunctatus (Diptera: Ceratopogonidae)

10.1101/2020.06.23.166496 ◽

2020 ◽

Author(s):

Jack Pilgrim ◽

Stefanos Siozios ◽

Matthew Baylis ◽

Gregory D. D. Hurst

Keyword(s):

Vertical Transmission ◽

Developmental Stages ◽

Fat Body ◽

Horizontal Transmission ◽

Follicle Cells ◽

Biting Midges ◽

Diverse Range ◽

Suspensory Ligament ◽

Transstadial Transmission ◽

Tissue Tropisms

AbstractRickettsia are a group of intracellular bacteria which can manipulate host reproduction and alter sensitivity to natural enemy attack in a diverse range of arthropods. The maintenance of Rickettsia endosymbionts in insect populations can be achieved through both vertical and horizontal transmission routes. For example, the presence of the symbiont in the follicle cells and salivary glands of Bemisia whiteflies allows Belli group Rickettsia transmission via the germline and plants, respectively. However, the transmission routes of other Rickettsia, such as those in the Torix group of the genus, remain underexplored. Through fluorescence in-situ hybridisation (FISH) and transmission electron microscopy (TEM) screening, this study describes the pattern of Torix Rickettsia tissue tropisms in the highland midge, Culicoides impunctatus (Diptera: Ceratopogonidae). Of note is high intensity of infection of the ovarian suspensory ligament, suggestive of a novel germline targeting strategy. Additionally, localisation of the symbiont in tissues of several developmental stages suggests transstadial transmission is a major route of ensuring maintenance of Rickettsia within C. impunctatus populations. Aside from providing insights into transmission strategies, Rickettsia presence in the fat body of larvae indicates potential host fitness and vector capacity impacts to be investigated in the future.Importance StatementMicrobial symbionts of disease vectors have garnered recent attention due to their ability to alter vectorial capacity. Their consideration as a means of arbovirus control depends on symbiont vertical transmission which leads to spread of the bacteria through a population. Previous work has identified a Rickettsia symbiont present in several vector species of biting midges (Culicoides spp.), however, symbiont transmission strategies and host effects remain underexplored. In this study, we describe the presence of Rickettsia in the ovarian suspensory ligament and the ovarian epithelial sheath of Culicoides impunctatus. Infection of these organs suggest the connective tissue surrounding developing eggs is important for ensuring vertical transmission of the symbiont in midges and possibly other insects. Additionally, our results indicate Rickettsia localisation in the fat body of Culicoides impunctatus. As viruses spread by midges often replicate in the fat body, this implies possible vector competence effects to be further investigated.

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Bacterial defenses against a natural antibiotic promote collateral resilience to clinical antibiotics

PLoS Biology ◽

10.1371/journal.pbio.3001093 ◽

2021 ◽

Vol 19 (3) ◽

pp. e3001093

Author(s):

Lucas A. Meirelles ◽

Elena K. Perry ◽

Megan Bergkessel ◽

Dianne K. Newman

Keyword(s):

Antibiotic Resistance ◽

Burkholderia Cepacia ◽

Evolutionary History ◽

Antibiotic Tolerance ◽

Human Pathogens ◽

Opportunistic Pathogens ◽

Diverse Range ◽

Natural Toxins ◽

Polymicrobial Communities ◽

Environmental Microbes

Bacterial opportunistic human pathogens frequently exhibit intrinsic antibiotic tolerance and resistance, resulting in infections that can be nearly impossible to eradicate. We asked whether this recalcitrance could be driven by these organisms’ evolutionary history as environmental microbes that engage in chemical warfare. Using Pseudomonas aeruginosa as a model, we demonstrate that the self-produced antibiotic pyocyanin (PYO) activates defenses that confer collateral tolerance specifically to structurally similar synthetic clinical antibiotics. Non-PYO-producing opportunistic pathogens, such as members of the Burkholderia cepacia complex, likewise display elevated antibiotic tolerance when cocultured with PYO-producing strains. Furthermore, by widening the population bottleneck that occurs during antibiotic selection and promoting the establishment of a more diverse range of mutant lineages, PYO increases apparent rates of mutation to antibiotic resistance to a degree that can rival clinically relevant hypermutator strains. Together, these results reveal an overlooked mechanism by which opportunistic pathogens that produce natural toxins can dramatically modulate the efficacy of clinical antibiotics and the evolution of antibiotic resistance, both for themselves and other members of clinically relevant polymicrobial communities.

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Community knowledge, perceptions, and practices regarding malaria and its control in Jabi Tehnan district, Amhara Region, Northwest Ethiopia

Malaria Journal ◽

10.1186/s12936-021-03996-5 ◽

2021 ◽

Vol 20 (1) ◽

Author(s):

Abebe Asale ◽

Zewdu Abro ◽

Bayu Enchalew ◽

Alayu Teshager ◽

Aklilu Belay ◽

...

Keyword(s):

Community Education ◽

Indoor Residual Spraying ◽

Northwest Ethiopia ◽

Primary Objective ◽

Malaria Episode ◽

Malaria Vectors ◽

Bed Nets ◽

Health Seeking ◽

District Health ◽

Malaria Parasites

Abstract Background Use of long-lasting insecticidal nets (LLINs), indoor residual spraying (IRS), community-based malaria education, prompt diagnosis and treatment are key programme components of malaria prevention and control in Ethiopia. However, the effectiveness of these interventions is often undermined by various challenges, including insecticide and drug resistance, the plasticity of malaria vectors feeding and biting behaviour, and certain household factors that lead to misuse and poor utilization of LLINs. The primary objective of this study was to document households’ perceptions towards malaria and assess the prevalence of the disease and the constraints related to the ongoing interventions in Ethiopia (LLINs, IRS, community mobilization house screening). Methods The study was conducted in Jabi Tehnan district, Northwestern Ethiopia, from November 2019 to March 2020. A total of 3010 households from 38 villages were randomly selected for socio-economic and demographic survey. Focus group discussions (FGDs) were conducted in 11 different health clusters considering agro-ecological differences. A total of 1256 children under 10 years of age were screened for malaria parasites using microscopy to determine malaria prevalence. Furthermore, 5-year malaria trend analysis was undertaken based on data obtained from the district health office to understand the disease dynamics. Results Malaria knowledge in the area was high as all FGD participants correctly identified mosquito bites during the night as sources of malaria transmission. Delayed health-seeking behaviour remains a key behavioural challenge in malaria control as it took patients on average 4 days before reporting the case at the nearby health facility. On average, households lost 2.53 working days per person-per malaria episode and they spent US$ 18 per person per episode. Out of the 1256 randomly selected under 10 children tested for malaria parasites, 11 (0.89%) were found to be positive. Malaria disproportionately affected the adult segment of the population more, with 50% of the total cases reported from households being from among individuals who were 15 years or older. The second most affected group was the age group between 5 and 14 years followed by children aged under 5, with 31% and 14% burden, respectively. Conclusion Despite the achievement of universal coverage in terms of LLINs access, utilization of vector control interventions in the area remained low. Using bed nets for unintended purposes remained a major challenge. Therefore, continued community education and communication work should be prioritized in the study area to bring about the desired behavioural changes.

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Impact of sugarcane irrigation on malaria vector Anopheles mosquito fauna, abundance and seasonality in Arjo-Didessa, Ethiopia

10.21203/rs.3.rs-37850/v3 ◽

2020 ◽

Author(s):

Assalif Demissew Shifera ◽

Dawit Hawaria ◽

Solomon Kibret ◽

Abebe Animut ◽

Arega Tsegaye ◽

...

Keyword(s):

Species Composition ◽

Malaria Vector ◽

Mosquito Species ◽

Malaria Vectors ◽

Anopheline Mosquito ◽

Chi Square ◽

Anopheles Species ◽

Mosquito Abundance ◽

Anopheles Mosquitoes ◽

The Impact

Abstract Background: Although irrigation activities are increasing in Ethiopia, limited studies evaluated their impact on malaria vector mosquito composition, abundance and seasonality. This study aimed at evaluating the impact of sugarcane irrigation on species composition, abundance and seasonality of malaria vectors. Methods : Adult Anopheles mosquitoes were collected using CDC light traps from three irrigated and three non-irrigated clusters in and around Arjo-Didessa sugarcane irrigation scheme in southwestern Ethiopia. Mosquito collections were conducted in four seasons: two wet and two dry, in 2018 and 2019. Mosquito species composition, abundance and seasonality were compared between irrigated and non-irrigated clusters. Anopheles mosquitoes were identified to species using morphological keys and An. gambiae s.l to sibling species using PCR. Chi-square was used to analyze the association between Anopheles species occurrence and environmental and seasonal parameters. Results: Overall, 2,108 female Anopheles mosquitoes comprising of six species were collected. Of these, 92.7% (n=1954) were from irrigated clusters and 7.3% (n=154) from the non-irrigated. An. gambiae s.l was the most abundant (67.3%) followed by An. coustani complex (25.3 %) and An. pharoensis (5.7%). PCR based identification revealed that 74.7% (n=168) of the An. gambiae s.l were An. arabiensis and 22.7% (n=51) An. amharicus . Density of An. gambiae s.l. (both indoor and outdoor) was higher in irrigated than non-irrigated clusters. The overall anopheline mosquito abundance during the wet seasons (87.2%; n=1837) was higher than the dry seasons (12.8%; n=271). Conclusion : The ongoing sugarcane irrigation activities in Arjo-Didessa created conditions suitable for increased malaria transmitting Anopheles species diversity and abundance. This in turn could drive malaria transmission in Arjo-Didessa and its environs in both dry and wet seasons. Thus, currently practiced malaria vector interventions need to be strengthened and consider larval source management to reduce vector abundance in the irrigated areas. Key Words: Malaria, Irrigation, Anopheles mosquitoes, vector density, An. amharicus , Ethiopia

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Malaria vectors in the Brazilian Amazon: Anopheles of the subgenus Nyssorhynchus

Revista do Instituto de Medicina Tropical de São Paulo ◽

10.1590/s0036-46652000000200005 ◽

2000 ◽

Vol 42 (2) ◽

pp. 87-94 ◽

Cited By ~ 140

Author(s):

Wanderli Pedro TADEI ◽

Bedsy DUTARY THATCHER

Keyword(s):

Sibling Species ◽

Activity Cycle ◽

Brazilian Amazon ◽

External Factors ◽

Malaria Vectors ◽

Human Intervention ◽

Principal Vector ◽

Anopheles Species ◽

Virgin Forest ◽

Continuous Activity

Various species of Anopheles (Nyssorhynchus) were studied in the Amazon with the objective of determining their importance as malaria vectors. Of the 33 known Anopheles species occurring in the Amazon, only 9 were found to be infected with Plasmodium. The different species of this subgenus varied both in diversity and density in the collection areas. The populations showed a tendency towards lower density and diversity in virgin forest than in areas modified by human intervention. The principal vector, An. darlingi, is anthropophilic with a continuous activity cycle lasting the entire night but peaking at sunset and sunrise. These species (Nyssorhynchus) are peridomiciliary, entering houses to feed on blood and immediately leaving to settle on nearby vegetation. Anopheles nuneztovari proved to be zoophilic, crepuscular and peridomiciliary. These habits may change depending on a series of external factors, especially those related to human activity. There is a possibility that sibling species exist in the study area and they are being studied with reference to An. darlingi, An. albitarsis and An. nuneztovari. The present results do not suggest the existence of subpopulations of An. darlingi in the Brazilian Amazon.

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