Laboratory Colony Research Articles

Oviposition Behavior of Culex quinquefasciatus and Anopheles coluzzii Females According to the Ovitrap Color and Presence of Fertilizer in Breeding Sites

Malaria and lymphatic filariasis are still important public health problems in Cameroon and in most part of sub-Saharan Africa. However there is still not enough informations on the bionomic and oviposition behaviour of their respectives vectors. The present study assesses the influence of color and organics pollutants presence on Culex and Anopheles coluzzii females oviposition behaviour. Laboratory experiments using Culex quinquefasciatus and Anopheles coluzzii laboratory colonies were conducted. Gravid females were offered cups of different colors (red, green, yellow, black, purple and white) with water for oviposition. Experiments were also conducted with cups containing different concentration of NPK fertilizers (1%, 0.75%, 0.5% and 0%). The influence of NPK fertilizer on the larval development time was also assessed. Cx quinquefasciatus and Anopheles coluzzii gravids females were found to be attracted by dark colors over 50% of eggs were laid in cups of black, red or green colors. The experiment was conducted with the mother and the F1 and F2 generations and similar oviposition preference were registered. When mosquitoes were offered cups containing different concentration of NPK for oviposition it appeared that females of the two species prefer laying their eggs in low concentration of NPK. Rearing of larvae in water containing NPK fertilizers was found to accelerate the larval development time of Culex quinquefasciaus but not of Anopheles coluzzii. The study indicated that several factors could be influencing vector bionomic and need further understanding to improve the fight against vector populations.

Age Polyethism in Atta sexdens (Linnaeus) (Hymenoptera: Formicidae)

This study aimed to verify age polyethism occurrence in medium-sized (cephalic capsule = 2.3 ± 0.21 mm) and small-sized (cc = 1.4 ± 0.10 mm) workers from Atta sexdens (Linnaeus) colonies. Four laboratory colonies were used, and they were maintained at 25 ± 2 °C, with 75 ± 3% relative humidity and a 12-hour photoperiod. Workers from these colonies were marked after their emergence and observed throughout their lifetime to determine which tasks they performed. The number of ants performing each activity was analyzed using linear mixed-effect models (LME), considering the temporal effect and the block design (colonies). We found that fungal garden maintenance tasks were frequent for both sizes, but their occurrence decreased significantly from the ninth week. The foraging activity occurred gradually in both sizes, with stabilization in the number of workers from the fourth week onwards and declined in the last three weeks of lifespan. Waste management tasks occurred throughout life but were more frequent during the first two weeks of life, in both medium and small workers. Therefore, age polyethism may be related to all activities; however, foraging tasks presented a distinct pattern compared to tasks in the fungus garden and refuse dump, where younger ants were more frequently observed.

Wolbachia strain wAlbB maintains high density and dengue inhibition following introduction into a field population of Aedes aegypti.

Aedes aegypti mosquitoes carrying the wAlbB Wolbachia strain show a reduced capacity to transmit dengue virus. wAlbB has been introduced into wild Ae. aegypti populations in several field sites in Kuala Lumpur, Malaysia, where it has persisted at high frequency for more than 2 years and significantly reduced dengue incidence. Although these encouraging results indicate that wAlbB releases can be an effective dengue control strategy, the long-term success depends on wAlbB maintaining high population frequencies and virus transmission inhibition, and both could be compromised by Wolbachia–host coevolution in the field. Here, wAlbB-carrying Ae. aegypti collected from the field 20 months after the cessation of releases showed no reduction in Wolbachia density or tissue distribution changes compared to a wAlbB laboratory colony. The wAlbB strain continued to induce complete unidirectional cytoplasmic incompatibility, showed perfect maternal transmission under laboratory conditions, and retained its capacity to inhibit dengue. Additionally, a field-collected wAlbB line was challenged with Malaysian dengue patient blood, and showed significant blocking of virus dissemination to the salivary glands. These results indicate that wAlbB continues to inhibit currently circulating strains of dengue in field populations of Ae. aegypti, and provides additional support for the continued scale-up of Wolbachia wAlbB releases for dengue control.This article is part of the theme issue ‘Novel control strategies for mosquito-borne diseases’.

Evolution of Wolbachia mutualism and reproductive parasitism: insight from two novel strains that co-infect cat fleas.

Wolbachiae are obligate intracellular bacteria that infect arthropods and certain nematodes. Usually maternally inherited, they may provision nutrients to (mutualism) or alter sexual biology of (reproductive parasitism) their invertebrate hosts. We report the assembly of closed genomes for two novel wolbachiae, wCfeT and wCfeJ, found co-infecting cat fleas (Ctenocephalides felis) of the Elward Laboratory colony (Soquel, CA, USA). wCfeT is basal to nearly all described Wolbachia supergroups, while wCfeJ is related to supergroups C, D and F. Both genomes contain laterally transferred genes that inform on the evolution of Wolbachia host associations. wCfeT carries the Biotin synthesis Operon of Obligate intracellular Microbes (BOOM); our analyses reveal five independent acquisitions of BOOM across the Wolbachia tree, indicating parallel evolution towards mutualism. Alternately, wCfeJ harbors a toxin-antidote operon analogous to the wPip cinAB operon recently characterized as an inducer of cytoplasmic incompatibility (CI) in flies. wCfeJ cinB and three adjacent genes are collectively similar to large modular toxins encoded in CI-like operons of certain Wolbachia strains and Rickettsia species, signifying that CI toxins streamline by fission of large modular toxins. Remarkably, the C. felis genome itself contains two CI-like antidote genes, divergent from wCfeJ cinA, revealing episodic reproductive parasitism in cat fleas and evidencing mobility of CI loci independent of WO-phage. Additional screening revealed predominant co-infection (wCfeT/wCfeJ) amongst C. felis colonies, though fleas in wild populations mostly harbor wCfeT alone. Collectively, genomes of wCfeT, wCfeJ, and their cat flea host supply instances of lateral gene transfers that could drive transitions between parasitism and mutualism.

Dormancy in laboratory-reared Asian longhorned beetles, Anoplophora glabripennis

An insect’s capacity to survive winter is critical for range expansion in temperate regions. The Asian longhorned beetle (Anoplophora glabripennis) is a polyphagous wood-boring insect native to China and the Korean peninsula and poses a high risk of invasion in North America and Europe. It is unclear whether A. glabripennis enters diapause, which means that diapause cannot be included in assessments of the risk of this species invading forests in temperate regions. Using a laboratory colony, we examine larval developmental arrest, metabolic rates, gas exchange patterns, thermal sensitivity, and body composition to characterize larval dormancy. Chilled larvae entered a temperature-independent developmental arrest which usually required more than four weeks of chilling to break, decreased their metabolic rate by as much as 63%, and maintained energy stores throughout the chilling period – results consistent with an obligate diapause. We also observed a switch to discontinuous gas exchange at low temperatures. Thermal sensitivity of metabolic rate did not differ between chilled and non-chilled larvae. Taken together, we conclude that A. glabripennis enters a larval diapause during chilling and terminates diapause after a requisite chilling period. These results will enhance our ability to predict phenology and potential distribution of current and future invasions of A. glabripennis.

Life cycle and larval growth of Dynamis borassi (Coleoptera: Dryophthoridae), an emerging pest to the peach palm

Dynamis borassi Fabricius (Coleoptera: Dryophthoridae) is widely distributed in South America and is an economic pest in commercially important palms (Arecaceae). In Colombia, peach palm production is of socio-economic importance in the Pacific region but has decreased dramatically due to a weevil complex that attacks the top of the palm. Dynamis borassi is the principal cause of this damage. Despite its importance, the biology of this species is unknown. A rigorous study of life cycle duration, as well as each stage of development, is key to future ecological and behavioural studies and essential in the design of comprehensive pest management plans. We report details of the life history of this pest in a laboratory colony. Life cycle and the larval growth were studied under controlled laboratory conditions (27 ± 1 °C and 75 ± 5% RH) using two artificial diets: Diet 1 without peach palm inflorescences and Diet 2 with 100 g of inflorescences. The egg stage lasted 3.5 ± 0.1 days, the larval stage 165.5 ± 4.7 days for Diet 1 and 153.8 ± 3.9 days for Diet 2, and the pupal stage 37 ± 1.7 days for Diet 1 and 39.7 ± 1.2 days for Diet 2. Twelve instars were observed. The instar when pupation took place varied from instar VII to XII. The weight gain curve had four growth phases with both diets. Each diet produced a different life cycle duration and growth rate. No significant differences were found in the life cycle between the diets.

German Culex pipiens biotype molestus and Culex torrentium are vector-competent for Usutu virus

BackgroundUsutu virus (USUV) is a rapidly spreading zoonotic arbovirus (arthropod-borne virus) and a considerable threat to the global avifauna and in isolated cases to human health. It is maintained in an enzootic cycle involving ornithophilic mosquitoes as vectors and birds as reservoir hosts. Despite massive die-offs in wild bird populations and the detection of severe neurological symptoms in infected humans, little is known about which mosquito species are involved in the propagation of USUV.MethodsIn the present study, the vector competence of a German (i.e. “Central European”) and a Serbian (i.e. “Southern European”) Culex pipiens biotype molestus laboratory colony was experimentally evaluated. For comparative purposes, Culex torrentium, a frequent species in Northern Europe, and Aedes aegypti, a primarily tropical species, were also tested. Adult female mosquitoes were exposed to bovine blood spiked with USUV Africa 2 and subsequently incubated at 25 °C. After 2 to 3 weeks saliva was collected from each individual mosquito to assess the ability of a mosquito species to transmit USUV.ResultsCulex pipiens biotype molestus mosquitoes originating from Germany and the Republic of Serbia and Cx. torrentium mosquitoes from Germany proved competent for USUV, as indicated by harboring viable virus in their saliva 21 days post infection. By contrast, Ae. aegypti mosquitoes were relatively refractory to an USUV infection, exhibiting low infection rates and lacking virus in their saliva.ConclusionsConsistent with the high prevalences and abundances of Cx. pipiens biotype molestus and Cx. torrentium in Central and Northern Europe, these two species have most likely played a historic role in the spread, maintenance, and introduction of USUV into Germany. Identification of the key USUV vectors enables the establishment and implementation of rigorous entomological surveillance programs and the development of effective, evidence-based vector control interventions.Graphical

MULTIPLEX POLYMERASE CHAIN REACTION ASSAY FOR THE DETECTION OF FIVE COMMON ENDOPARASITE INFESTATIONS IN LABORATORY RODENTS

Rodent pinworms, Spironucleus muris and Tritrichomonas muris are the endoparasites that should be monitored and excluded from laboratory animal colonies. Nevertheless, traditional diagnostic methods may not efficiently detect and accurately demonstrate the endoparasite infestation status. In this study, we developed a multiplex PCR assay targeting the rRNA genes to simultaneously detect and differentiate five endoparasites, including Syphacia obvelata, Syphacia muris, Aspiculuris tetraptera, Spironucleus muris, and T. muris, as well as a housekeeping gene in feces. The multiplex PCR could identify an equivalent infection of pinworm, Spironucleus muris and T. muris, with a detection limit of as few as 10 copies. Furthermore, dual infections with up to 100-fold differences and triple infections with 10-fold differences in parasite loads can also be detected. In comparison of traditional methods with the multiplex PCR assay, 76 rodents from 11 research colonies and 3 pet shops and additional 27 fecal samples from laboratory rodents were screened for the infestation status of the five endoparasites. The multiplex PCR had higher sensitivity (97.2–100%) and accuracy (99–100%) than those of the traditional antemortem (sensitivity: 83–100%; accuracy: 94–100%) and postmortem methods (sensitivity: 75–100%; accuracy: 92.1–100%). In addition, an early stage of S. obvelata contamination in a SPF laboratory animal colony was also successfully detected by this multiplex PCR assay. This Pinworm/Spironucleus/Tritrichomonas/Actin Multiplex PCR assay should be a powerful tool to screen endoparasite infestations in laboratory colonies without animal sacrifice.

Influence of Temperature on the Life-Cycle Dynamics of Aedes albopictus Population Established at Temperate Latitudes: A Laboratory Experiment

Simple SummaryMosquitoes represent a potential major public health concern, as they are capable of transmitting several pathogens when biting humans. It is well known that temperature is a crucial factor affecting mosquito biology: for instance, warmer conditions can increase survival and fecundity. Here, we quantify the influence of different temperatures on the bionomics of Aedes albopictus, which is a mosquito species native to Southeast Asia that has been able to spread worldwide during the last forty years. We used specimens collected from northern Italy to assess if temperate individuals are characterized, possibly thanks to an adaptation process, by a different thermal response with respect to subtropical individuals. We found that immature stages are well adapted to colder temperatures, which nonetheless seem to prevent any blood-feeding activity. Adult longevity and fecundity were substantially greater at mild conditions. This thermal adaptation might increase the length of the breeding season and could allow the colonization of areas at higher altitude, resulting in an overall increased risk for potential transmission of Ae. albopictus-borne pathogens.The mosquito species Aedes albopictus has successfully colonized many areas at temperate latitudes, representing a major public health concern. As mosquito bionomics is critically affected by temperature, we experimentally investigated the influence of different constant rearing temperatures (10, 15, 25, and 30 °C) on the survival rates, fecundity, and developmental times of different life stages of Ae. albopictus using a laboratory colony established from specimens collected in northern Italy. We compared our results with previously published data obtained with subtropical populations. We found that temperate Ae. albopictus immature stages are better adapted to colder temperatures: temperate larvae were able to develop even at 10 °C and at 15 °C, larval survivorship was comparable to the one observed at warmer conditions. Nonetheless, at these lower temperatures, we did not observe any blood-feeding activity. Adult longevity and fecundity were substantially greater at 25 °C with respect to the other tested temperatures. Our findings highlight the ability of Ae. albopictus to quickly adapt to colder environments and provide new important insights on the bionomics of this species at temperate latitudes.

A Novel Anphevirus in Aedes albopictus Mosquitoes Is Distributed Worldwide and Interacts with the Host RNA Interference Pathway.

The Asian tiger mosquito Aedes albopictus is a competent vector for several human arboviruses including dengue, chikungunya and Zika viruses. Mosquitoes also harbor insect-specific viruses (ISVs) that may modulate host physiology and potentially affect the transmission of viruses that are pathogenic to vertebrates, thus representing a potential tool for vector control strategies. In Ae. albopictus we identified a novel anphevirus (family Xinmoviridae; order Mononegavirales) provisionally designated here as Aedes albopictus anphevirus (AealbAV). AealbAV contains a ~12.4 kb genome that is highly divergent from currently known viruses but displays gene content and genomic organization typical of known anpheviruses. We identified AealbAV in several publicly available RNA-Seq datasets from different geographical regions both in laboratory colonies and field collected mosquitoes. Coding-complete genomes of AealbAV strains are highly similar worldwide (>96% nucleotide identity) and cluster according to the geographical origin of their hosts. AealbAV appears to be present in various body compartments and mosquito life stages, including eggs. We further detected AealbAV-derived vsiRNAs and vpiRNAs in publicly available miRNA-Seq libraries of Ae. albopictus and in samples experimentally coinfected with chikungunya virus. This suggests that AealbAV is targeted by the host RNA interference (RNAi) response, consistent with persistent virus replication. The discovery and characterization of AealbAV in Ae. albopictus will now allow us to identify its infection in mosquito populations and laboratory strains, and to assess its potential impact on Ae. albopictus physiology and ability to transmit arboviruses.

Sampling Efficacy and Survival Rates of Labarrus pseudolividus (Coleoptera: Scarabaeidae) and Onthophagus taurus (Coleoptera: Scarabaeidae) Using Flotation and Sieve-Separation Methodology.

Understanding collection methodologies and their limitations are essential when targeting specific arthropods for use in habitat restoration, conservation, laboratory colony formation, or when holistically representing local populations using ecological surveys. For dung beetles, the most popular collection methodology is baited traps, followed by light traps and unbaited flight-intercept traps during diversity surveys. A less common collection method, flotation, is assumed to be laborious and messy, and so only a handful of papers exist on its refinement and strengths. Our purpose was threefold: First, we tested the recovery and survival rates of Labarrus (=Aphodius) pseudolividus (Balthasar) and Onthophagus taurus (Schreber) when floating beetle-seeded dung pats to determine potential collection and safety issues. We collected 72.4 and 78% of the seeded L. pseudolividus and O. taurus, respectively, with >95% survival rating. Second, we developed a flotation-sieving technique that enables users to rapidly collect and passively sort dung beetles with less time and effort. Specifically, we often collected 50–100 g of wild dung beetles within a couple of hours of gathering dung and sorted them in a couple more by allowing dung beetles to sort themselves by size within a series of sieves; Third, we reviewed flotation-based advantages and disadvantages in comparison to other methodologies.

External female genitalia of Triatoma jatai, Triatoma costalimai and Triatoma williami (Hemiptera: Reduviidae: Triatominae)

BackgroundTaxonomic identification of triatomines is generally performed based on aspects of their external morphology. However, the use of a multidisciplinary approach, considering morphological aspects of the external genitalia, morphometry, genetics, and phylogeography has been suggested, especially for similar and/or cryptic species. The rupestral species Triatoma jatai Gonçalves et al., 2013, Triatoma costalimai Verano & Galvão, 1959 and Triatoma williami Galvão et al., 1965, which are morphologically similar, have been found naturally infected with Trypanosoma cruzi (Chagas, 1909) in wild, peridomestic, and intradomestic environments, representing a risk of new outbreaks of Chagas disease. This study presents morphological description complementation of these species, with an emphasis on the structures of the female external genitalia, using scanning electron microscopy.MethodsThe females of T. jatai and T. costalimai (n = 10 of each) were captured in the Brazilian municipalities of Paranã and Aurora do Tocantins and were identified with the use of a dichotomous key for the Matogrossensis subcomplex. Females of T. williami (n = 5), were obtained from a laboratory colony. The females were cut transversely at the sixth abdominal segment and examined under scanning electron microscopy (SEM) at the Oswaldo Cruz/Fiocruz Institute Electronic Microscopy Platform.ResultsIt was possible to differentiate the three species based on the characteristics of urotergites VII, VIII and IX and urosternite VII, as well as the genital plaques, gonocoxites, and gonapophyses. To our knowledge, morphological differences in the spines present on gonapophysis 8 in triatomines are described here for the first time.ConclusionsThe results show that external genitalia of females are useful structures to differentiate T. costalimai, T. jatai and T. williami. SEM analysis contributes to and corroborates, together with other tools morphological and molecular, the distinction of the three species.

Performance of Ooencyrtus kuvanae (Hymenoptera: Encyrtidae) on two host species, Halyomorpha halys and Philosamia ricini

AbstractThe brown marmorated stink bug, Halyomorpha halys, is an invasive agricultural pest of fruit trees and vegetables. Egg parasitoids play a key role in the reducing of H. halys populations. Ooencyrtus kuvanae (Howard) (Hymenoptera: Encyrtidae) can parasitize H. halys and complete its life cycle in this host species. Many factors can influence this parasitoid–host relationship. Of these factors, we evaluated the effect of female age, exposure time, and host species on the biological characteristics and fecundity of O. kuvanae reared on eggs of H. halys as well as another previously known host Philosamia ricini (Lepidoptera: Saturniidae). In this study, we used a 3‐year‐old laboratory colony of O. kuvanae. Parasitism rates positively affected by exposure time in P. ricini. The highest parasitism rates were obtained in 5‐ and 7‐day‐old females of both hosts. The highest emergence rates were recorded on P. ricini for 5‐ and 7‐day‐old female P. ricini (81.8% and 84.8%, respectively). The development time of O. kuvanae ranged from 18.4 to 19.1 days on H. halys and 17.7 to 18.3 days on P. ricini. The longevity of O. kuvanae that were provided honey was 38.5 and 47.8 days on H. halys and P. ricini, respectively. The longevity of O. kuvanae that were not provided honey was 2.3 and 2.8 days on H. halys and P. ricini, respectively. The sex ratio was male‐biased (36.5% female) on H. halys and female‐biased (55.2% female) on P. ricini. Fecundity of O. kuvanae was 37.7 and 59.6 progeny per female for H. halys and P. ricini, respectively. The performance of O. kuvanae was lower when compared with its performance on the host P. ricini. Our results suggest that O. kuvanae has potential as new biological control agent for H. halys.

Cannibalism, oviposition and egg development in the edible long-horned grasshopper, Ruspolia differens (Orthoptera: Tettigoniidae) under laboratory conditions

Reliance on seasonal wild harvests of the edible long-horned grasshopper Ruspolia differens (Serville) (Orthoptera: Tettigoniidae) needs to be broken through developing efficient mass rearing protocols. This study aimed at boosting productivity of laboratory colonies of R. differens through understanding ways of minimising cannibalism, selecting suitable oviposition substrates and enhancing egg development and hatchability. Firstly, we investigated the extent of cannibalism in a colony of R. differens relative to total mortality, sex, diurnality, growth stage, body parts eaten and visual barriers (presence/absence of cardboard egg trays). R. differens cadavers recovered from cages with/without the egg trays were scored for cause of death (cannibalism/other), time of death (night/day), sex, growth stage and body part devoured. Secondly, cotton wool and leaf sheaths of Pennisetum sp., maize and Panicum sp. were evaluated for oviposition preference by R. differens. Eggs oviposited in each substrate were incubated to determine developmental duration and hatchability. Thirdly, we tested effect of moistening R. differens eggs on incubation period and hatchability in intact and opened leaf sheaths against unmoistened eggs in intact sheaths as a control. We found that cannibalism accounted for 49% of R. differens deaths with 83% of victims being adults. Males and females were equally susceptible to cannibalism. Over 97% of cannibalism occurred at night and presence of egg trays almost doubled incidence of cannibalism. The thorax was the most preferred body part, which was consumed in 77% of cannibalised individuals. Maize and Panicum sp. were preferred for oviposition over Pennisetum sp. and cotton wool. Opening of egg-laden leaf sheaths reduced incubation period by 0.4 days. Unmoistened eggs didn’t hatch after incubation for 25 days, but 65% of them hatched within 11 days upon moistening. These findings offer guidelines for improvement of protocols for mass rearing of R. differens for human consumption and other uses.

Partitiviruses Infecting Drosophila melanogaster and Aedes aegypti Exhibit Efficient Biparental Vertical Transmission.

Partitiviruses are segmented, multipartite double-stranded RNA (dsRNA) viruses that until recently were only known to infect fungi, plants, and protozoans. Metagenomic surveys have revealed that partitivirus-like sequences are also commonly associated with arthropods. One arthropod-associated partitivirus, galbut virus, is common in wild populations of Drosophila melanogaster To begin to understand the processes that underlie this virus's high global prevalence, we established colonies of wild-caught infected flies. Infection remained at stably high levels over 3 years, with between 63 and 100% of individual flies infected. Galbut virus infects fly cells and replicates in tissues throughout infected adults, including reproductive tissues and the gut epithelium. We detected no evidence of horizontal transmission via ingestion, but vertical transmission from either infected females or infected males was ∼100% efficient. Vertical transmission of a related partitivirus, verdadero virus, that we discovered in a laboratory colony of Aedes aegypti mosquitoes was similarly efficient. This suggests that efficient biparental vertical transmission may be a feature of at least a subset of insect-infecting partitiviruses. To study the impact of galbut virus infection free from the confounding effect of other viruses, we generated an inbred line of flies with galbut virus as the only detectable virus infection. We were able to transmit infection experimentally via microinjection of homogenate from these galbut-only flies. This sets the stage for experiments to understand the biological impact and possible utility of partitiviruses infecting model organisms and disease vectors.IMPORTANCE Galbut virus is a recently discovered partitivirus that is extraordinarily common in wild populations of the model organism Drosophila melanogaster Like for most viruses discovered through metagenomics, most of the basic biological questions about this virus remain unanswered. We found that galbut virus, along with a closely related partitivirus found in Aedes aegypti mosquitoes, is transmitted from infected females or males to offspring with ∼100% efficiency and can be maintained in laboratory colonies over years. This efficient transmission mechanism likely underlies the successful spread of these viruses through insect populations. We created Drosophila lines that contained galbut virus as the only virus infection and showed that these flies can be used as a source for experimental infections. This provides insight into how arthropod-infecting partitiviruses may be maintained in nature and sets the stage for exploration of their biology and potential utility.

Stability of the Virome in Lab- and Field-Collected Aedes albopictus Mosquitoes across Different Developmental Stages and Possible Core Viruses in the Publicly Available Virome Data of Aedes Mosquitoes.

Aedes mosquitoes can efficiently transmit many pathogenic arboviruses, placing a great burden on public health worldwide. In addition, they also carry a number of insect-specific viruses (ISVs), and it was recently suggested that some of these ISVs might form a stable species-specific "core virome" in mosquito populations. However, little is known about such a core virome in laboratory colonies and if it is present across different developmental stages. In this study, we compared the viromes in eggs, larvae, pupae, and adults of Aedes albopictus mosquitoes collected from a lab colony and compared each to the virome of different developmental stages collected in the field. The virome in lab-derived A. albopictus was very stable across all stages, consistent with a vertical transmission route of these viruses, and formed a possible "vertically transmitted core virome." The different stages of field-collected A. albopictus mosquitoes also contained this stable vertically transmitted core virome, as well as another set of viruses (e.g., viruses distantly related to Guadeloupe mosquito virus, Hubei virga-like virus 2, and Sarawak virus) shared by mosquitoes across different stages, which might represent an "environment-derived core virome." To further study this core set of ISVs, we screened 48 publicly available SRA viral metagenomic data sets of mosquitoes belonging to the genus Aedes, showing that some of the identified ISVs were identified in the majority of SRAs and providing further evidence supporting the core-virome concept.IMPORTANCE Our study revealed that the virome was very stable across all developmental stages of both lab-derived and field-collected Aedes albopictus The data representing the core virome in lab A. albopictus proved the vertical transmission route of these viruses, forming a "vertically transmitted core virome." Field mosquitoes also contained this stable vertically transmitted core virome as well as additional viruses, which probably represented "environment-derived core virome" and which therefore were less stable over time and geography. By further screening publicly available SRA viral metagenomic data sets from mosquitoes belonging to the genus Aedes, some of the identified core ISVs were shown to be present in the majority of SRAs, such as Phasi Charoen-like phasivirus and Guadeloupe mosquito virus. How these core ISVs influence the biology of the mosquito host and arbovirus infection and evolution deserves to be further explored.

Assessment of the Hemotek® system for the in vitro feeding of field-collected Culicoides imicola (Diptera: Ceratopogonidae) in South Africa.

The optimising and standardisation of in vitro blood feeding protocols for field-collected Culicoides species (Diptera: Ceratopogonidae) will be of essence for the comparison of the vector competencies of various populations of viruses of veterinary importance and the establishment of laboratory colonies of putative vector species. A custom-made feeding chamber to accommodate the small size of Culicoides imicola Kieffer was designed for the commercially available Hemotek® system and compared to existing membrane and cotton pledge feeding methods. High feeding rates coupled to higher mean blood meal volume than that of the existing OVI device indicated that the Hemotek system will be suitable for the feeding of field-collected Culicoides. The Hemotek system was subsequently used to identify factors that may affect feeding success in the laboratory. Evaluated factors were the source (host) and temperature of the blood meal, time of the day of feeding, the position of the blood reservoir in relation to the midges and exposure time to the blood. While only feeding orientation and the temperature of the blood source seems to significantly affect the feeding rate, all the factors did influence the volume of blood consumed.

Inter‐colony invasion between wild naked mole‐rat colonies

AbstractDespite published reports of inter‐colony aggression and invasion in the laboratory, direct aggressive competition for resources has not previously been considered as a driving force for the large colony size in naked mole‐rats (Heterocephalus glaber). We report evidence that wild colonies of naked mole‐rats invade neighbouring colonies and thus expand their territories. Furthermore, as has been observed in laboratory colonies, invading colonies may kidnap un‐weaned pups which can subsequently be incorporated into the workforce. This is convergent with the slave‐making ants and suggests that intergroup aggression between naked mole‐rat colonies may be a strong selective pressure for group living and extreme colony size.

Identification and genome sequencing of RNA viruses in the eucalyptus snout beetle Gonipterus spp. (Coleoptera: Curculionidae).

The genomes of two putative new RNA viruses (macula-like virus and bunya-like virus) were identified in total RNA extracted from dead eucalyptus snout beetles (Gonipterus spp.) from a laboratory colony. However, only bunya-like virus was detected in field-collected insects. The macula-like virus has a monopartite single-stranded RNA genome that contains three open reading frames (ORFs) encoding an RNA-dependent RNA polymerase (RdRp), a capsid protein (CP), protein with unknown function. The bunya-like virus genome was predicted to consist of two RNA segments: a large segment (L) encoding a single protein (RdRp) and a small segment (S) encoding a putative nucleocapsid protein.

Considerations for Human Blood-Feeding and Arthropod Exposure in Vector Biology Research: An Essential Tool for Investigations and Disease Control.

Eventually there may be a broadly acceptable, even perfected, substitute for the human host requirement for direct feeding experiments by arthropods, most notably mosquitoes. However, for now, direct and indirect feeding on human volunteers is an important, if not essential, tool in vector biology research (VBR). This article builds on the foundational publication by Achee et al. (2015) covering considerations for the use of human participants in VBR pursuits. The authors introduced methods involving human participation in VBR, while detailing human-landing collections (catches) as a prime example. Benedict et al. (2018) continued this theme with an overview of human participation and considerations for research that involves release of mosquito vectors into the environment. In this study, we discuss another important aspect of human use in VBR activities: considerations addressing studies that require an arthropod to feed on a live human host. Using mosquito studies as our principal example, in this study, we discuss the tremendous importance and value of this approach to support and allow study of a wide variety of factors and interactions related to our understanding of vector-borne diseases and their control. This includes establishment of laboratory colonies for test populations, characterization of essential nutrients that contribute to mosquito fitness, characterization of blood-feeding (biting) behavior and pathogen transmission, parameterization for modeling transmission dynamics, evaluation of human host attraction and/or agents that repel, and the effectiveness of antivector or parasite therapeutic drug studies.