Laboratory maintenance and care of Rhodnius prolixus (Hemiptera: Reduviidae) and other Triatominae.

Triatomines (Hemiptera: Reduviidae) are hematophagous insects that transmit Trypanosoma cruzi, the etiological agent for Chagas disease, to humans and other mammals. As medically important vectors, species such as Rhodnius prolixus (Hemiptera: Reduviidae) have long been used as a model organism for physiological studies. Laboratory rearing of triatomines is needed to support vector and parasite research.

Juvenile Hormone as a contributing factor in establishing midgut microbiota for fecundity and fitness enhancement in adult female Aedes aegypti.

Understanding the factors influencing mosquitoes' fecundity and longevity is important for designing better and more sustainable vector control strategies, as these parameters can impact their vectorial capacity. Here, we address how mating affects midgut growth in Aedes aegypti, what role Juvenile Hormone (JH) plays in this process, and how it impacts the mosquito's immune response and microbiota. Our findings reveal that mating and JH induce midgut growth.

Adult Anesthesia, Feeding, and Sex Separation.

The adult stage is the only nonaquatic stage of the mosquito. Both male and female mosquitoes require access to a source of sugar to survive. In the insectary, a temperature of ∼27°C and 80% relative humidity and a cycle of 12 h light:12 h dark light, ideally with a sunrise and sunset period, are necessary minimum conditions to mimic their natural environment.

Larval Rearing.

Mosquito larvae are aquatic and go through four development stages (larval instars L1-L4) before pupation. Species vary in the duration of larval development, and a variety of external factors affect the development rate (e.g.

Considerations for Rearing and Maintaining in the Laboratory.

mosquitoes can transmit several human pathogens, including viruses such as o'nyong-nyong and parasites including spp. and spp., which cause malaria and filariasis, respectively.

Egg Collection, Disinfection, and Hatching.

Gravid (i.e., with fully developed eggs), mated females typically lay their eggs directly on water ∼48-72 h after a blood meal.

Pupa Collection and Sex Identification.

For most species, larval-pupal metamorphosis commences ∼1 wk after egg hatching. However, depending on the amount of food provided, HO temperature, and larval density, the pupation process can be accelerated or delayed. Synchronous pupation is difficult to accomplish consistently, and, thus, pupae need to be separated from larvae daily.

Evaluation of the residual efficacy and physical durability of five long-lasting insecticidal nets (LLINs) in Senegal.

Background: The preventive and curative strategies of malaria are based on promoting the use of long-lasting insecticidal nets (LLINs) and treating confirmed cases with artemisinin-based combination therapy. These strategies have led to a sharp decline in the burden of malaria, which remains a significant public health problem in sub-Saharan countries. The objective of this study was to determine and compare the residual efficacy of LLINs recommended by the World Health Organization.

Adaptation of ELISA detection of Plasmodium falciparum and Plasmodium vivax circumsporozoite proteins in mosquitoes to a multiplex bead-based immunoassay.

Background: Plasmodium spp. sporozoite rates in mosquitoes are used to better understand malaria transmission intensity, the relative importance of vector species and the impact of interventions. These rates are typically estimated using an enzyme-linked immunosorbent assay (ELISA) utilizing antibodies against the circumsporozoite protein of Plasmodium falciparum, Plasmodium vivax VK210 (P.

Delivery of Double-Stranded RNAs (dsRNAs) Produced by Escherichia coli HT115(DE3) for Nontransgenic RNAi-Based Insect Pest Management.

RNA interference (RNAi) is a powerful mechanism that can be exploited not only for physiology research but also for designing insect pest management approaches. Some insects cause harm by vectoring diseases dangerous to humans, livestock, or plants or by damaging crops. For at least a decade now, different insect control strategies that induce RNAi by delivering double stranded RNA (dsRNA) targeting essential genes have been proposed.

Surviving the Journey: Comparisons of Temperature-Stabilizing Materials for Living Arthropod Shipments.

Shipments of living mosquitoes and other arthropods require temperatures that are within a range that is compatible with their health and survival. In addition to express shipping and insulated containers, shipments often include materials that either store heat (i.e.

Cryoprotectant toxicity and hypothermic sensitivity among Anopheles larvae.

Laboratory rearing of mosquitoes is commonly practiced by researchers studying mosquito-borne infectious diseases and vector control methods. In the absence of cryopreservation methods to stabilize unique or genetically modified strains, mosquito lines must be continuously maintained, a laborious process that risks selection effects, contamination, and genetic drift. Towards the development of a cryopreservation protocol, several commonly used cryoprotectants were systematically characterized here both individually and as cocktails.

Delimiting cryptic morphological variation among human malaria vector species using convolutional neural networks.

Deep learning is a powerful approach for distinguishing classes of images, and there is a growing interest in applying these methods to delimit species, particularly in the identification of mosquito vectors. Visual identification of mosquito species is the foundation of mosquito-borne disease surveillance and management, but can be hindered by cryptic morphological variation in mosquito vector species complexes such as the malaria-transmitting Anopheles gambiae complex. We sought to apply Convolutional Neural Networks (CNNs) to images of mosquitoes as a proof-of-concept to determine the feasibility of automatic classification of mosquito sex, genus, species, and strains using whole-body, 2D images of mosquitoes.

Physiological responses to cryoprotectant treatment in an early larval stage of the malaria mosquito, Anopheles gambiae.

The development of cryopreservation protocols for Anopheles gambiae could significantly improve research and control efforts. Cryopreservation of any An. gambiae life stage has yet to be successful.

Trials of the Automated Particle Counter for laboratory rearing of mosquito larvae.

As a means of obtaining reproducible and accurate numbers of larvae for laboratory rearing, we tested a large-particle flow-cytometer type device called the 'Automated Particle Counter' (APC). The APC is a gravity-fed, self-contained unit that detects changes in light intensity caused by larvae passing the detector in a water stream and controls dispensing by stopping the flow when the desired number has been reached. We determined the accuracy (number dispensed compared to the target value) and precision (distribution of number dispensed) of dispensing at a variety of counting sensitivity thresholds and larva throughput rates (larvae per second) using < 1-day old Anopheles gambiae and Aedes aegypti larvae.

Fluorescent markers rhodamine B and uranine for Anopheles gambiae adults and matings.

Background: Marking mosquitoes is vital for mark-release-recapture and many laboratory studies, but their small size precludes the use of methods that are available for larger animals such as unique identifier tags and radio devices. Fluorescent dust is the most commonly used method to distinguish released individuals from the wild population. Numerous colours and combinations can be used, however, dust sometimes affects longevity and behaviour so alternatives that do not have these effects would contribute substantially.

An Atypical Case of Autochthonous Cutaneous Leishmaniasis Associated with Naturally Infected Phlebotomine Sand Flies in Texas, United States.

In the United States, phlebotomine sand flies carrying () are endemic along the southern border. However, relatively little is known about the enzootic and zoonotic transmission of () within the United States, and autochthonous cases of the consequent disease are rarely reported. We investigated an atypical case of cutaneous leishmaniasis (CL) caused by () in a patient from central Texas which did not respond to a typical antileishmanial chemotherapy.

Pragmatic selection of larval mosquito diets for insectary rearing of Anopheles gambiae and Aedes aegypti.

Larval mosquitoes are aquatic omnivorous scavengers which scrape food from submerged surfaces and collect suspended food particles with their mouth brushes. The composition of diets that have been used in insectaries varies widely though necessarily provides sufficient nutrition to allow colonies to be maintained. Issues such as cost, availability and experience influence which diet is selected.

Downregulation of female doublesex expression by oral-mediated RNA interference reduces number and fitness of Anopheles gambiae adult females.

Background: Mosquito-borne diseases affect millions worldwide, with malaria alone killing over 400 thousand people per year and affecting hundreds of millions. To date, the best strategy to prevent the disease remains insecticide-based mosquito control. However, insecticide resistance as well as economic and social factors reduce the effectiveness of the current methodologies.

Plasmodium falciparum (Haemosporodia: Plasmodiidae) and O'nyong-nyong Virus Development in a Transgenic Anopheles gambiae (Diptera: Culicidae) Strain.

Transgenic Anopheles gambiae Giles (Diptera: Culicidae) mosquitoes have been developed that confer sexual sterility on males that carry a transgene encoding a protein which cuts ribosomal DNA. A relevant risk concern with transgenic mosquitoes is that their capacity to transmit known pathogens could be greater than the unmodified form. In this study, the ability to develop two human pathogens in these transgenic mosquitoes carrying a homing endonuclease which is expressed in the testes was compared with its nontransgenic siblings.

Infection prevention and control training and capacity building during the Ebola epidemic in Guinea.

Background: During the 2014-2016 Ebola epidemic in West Africa, a key epidemiological feature was disease transmission within healthcare facilities, indicating a need for infection prevention and control (IPC) training and support.

Methods: IPC training was provided to frontline healthcare workers (HCW) in healthcare facilities that were not Ebola treatment units, as well as to IPC trainers and IPC supervisors placed in healthcare facilities. Trainings included both didactic and hands-on components, and were assessed using pre-tests, post-tests and practical evaluations.

Pioneer study of population genetics of Rhodnius ecuadoriensis (Hemiptera: Reduviidae) from the central coastand southern Andean regions of Ecuador.

Effective control of Chagas disease vector populations requires a good understanding of the epidemiological components, including a reliable analysis of the genetic structure of vector populations. Rhodnius ecuadoriensis is the most widespread vector of Chagas disease in Ecuador, occupying domestic, peridomestic and sylvatic habitats. It is widely distributed in the central coast and southern highlands regions of Ecuador, two very different regions in terms of bio-geographical characteristics.

Malaria vector research and control in Haiti: a systematic review.

Background: Haiti has a set a target of eliminating malaria by 2020. However, information on malaria vector research in Haiti is not well known. This paper presents results from a systematic review of the literature on malaria vector research, bionomics and control in Haiti.

What Do We Know About Chagas Disease in the United States?

Chagas disease, caused by the parasite Trypanosoma cruzi, affects more than 5 million people worldwide leading to serious heart and gastrointestinal disease in a proportion of chronically infected patients. Important modes of transmission include vector-borne, congenital, and via blood transfusion or organ transplant from an infected donor. Vector-borne transmission of Chagas disease occurs in the Americas, including the southern half of North America, where the specific vector insects (triatomines), T.