Harvesting insect pests for animal feed: potential to capture an unexploited resource.

The demand for animal protein grows as the human population increases. Technological and genetic advances in traditional animal agriculture will not produce enough protein to meet future needs without significant innovations such as the use of insects as protein sources. Insect farming is growing insects, whereas insect harvesting is collecting insects from their natural habitats to produce high-quality protein for animal feed or human food. Intensive agricultural environments produce tremendous quantities of pestiferous insects and with the right harvest technologies these insects can be used as a protein supplement in traditional animal daily rations. An avenue to exploit these insects is to use traps such as the United States Department of Agriculture-Biomass Harvest Trap (USDA-BHT) to efficiently attract, harvest, and store insects from naturally abundant agricultural settings. The modular design allows for a low cost, easy to build and fix device that is user friendly and has customizable attractants to target various pest species. Although insect harvesting faces substantial challenges, including insect biomass quantity, seasonal abundance and preservation, food safety, and economic and nutritional evaluation, the potential for utilizing these pests for protein shows tremendous promise. In this forum, insect harvesting is discussed, including its potential, limitations, challenges, and research needs. In addition, the use of a mass trapping device is discussed as a tool to increase the biomass of insects collected from the environment.

Conceptualization, design, and construction of a novel insect mass trapping device: the USDA Biomass Harvest Trap (USDA-BHT).

The use of insects as animal feed has the potential to be a green revolution for animal agriculture as insects are a rich source of high-quality protein. Insect farming must overcome challenges such as product affordability and scalability before it can be widely incorporated as animal feed. An alternative is to harvest insect pests from the environment using mass trapping devices and use them as animal feed. For example, intensive agricultural environments generate large quantities of pestiferous insects and with the right harvest technologies, these insects can be used as a protein supplement in traditional animal daily rations. Most insect trapping devices are limited by the biomass they can collect. In that context, and with the goal of using wild collected insects as animal feed, the United States Department of Agriculture-Biomass Harvest Trap (USDA-BHT) was designed and built. The USDA-BHT is a valuable mass trapping device developed to efficiently attract, harvest, and store flying insects from naturally abundant agricultural settings. The trap offers a modular design with adjustable capabilities, and it is an inexpensive device that can easily be built with commonly available parts and tools. The USDA-BHT is also user-friendly and has customizable attractants to target various pest species.

Assessment of the USDA Biomass Harvest Trap (USDA-BHT) device as an insect harvest and mosquito surveillance tool.

Insects are a promising source of high-quality protein, and the insect farming industry will lead to higher sustainability when it overcomes scaling up, cost effectiveness, and automation. In contrast to insect farming (raising and breeding insects as livestock), wild insect harvesting (collecting agricultural insect pests), may constitute a simple sustainable animal protein supplementation strategy. For wild harvest to be successful sufficient insect biomass needs to be collected while simultaneously avoiding the collection of nontarget insects. We assessed the performance of the USDA Biomass Harvest Trap (USDA-BHT) device to collect flying insect biomass and as a mosquito surveillance tool. The USDA-BHT device was compared to other suction traps commonly used for mosquito surveillance (Centers for Disease Control and Prevention (CDC) light traps, Encephalitis virus surveillance traps, and Biogents Sentinel traps). The insect biomass harvested in the USDA-BHT was statistically higher than the one harvested in the other traps, however the mosquito collections between traps were not statistically significantly different. The USDA-BHT collected some beneficial insects, although it was observed that their collection was minimized at night. These findings coupled with the fact that sorting time to separate the mosquitoes from the other collected insects was significantly longer for the USDA-BHT, indicate that the use of this device for insect biomass collection conflicts with its use as an efficient mosquito surveillance tool. Nevertheless, the device efficiently collected insect biomass, and thus can be used to generate an alternative protein source for animal feed.

The microbiota of Amblyomma americanum reflects known westward expansion.

Amblyomma americanum, a known vector of multiple tick-borne pathogens, has expanded its geographic distribution across the United States in the past decades. Tick microbiomes may play a role shaping their host's life history and vectorial capacity. Bacterial communities associated with A. americanum may reflect, or enable, geographic expansion and studying the microbiota will improve understanding of tick-borne disease ecology. We examined the microbiota structure of 189 adult ticks collected in four regions encompassing their historical and current geographic distribution. Both geographic region of origin and sex were significant predictors of alpha diversity. As in other tick models, within-sample diversity was low and uneven given the presence of dominant endosymbionts. Beta diversity analyses revealed that bacterial profiles of ticks of both sexes collected in the West were significantly different from those of the Historic range. Biomarkers were identified for all regions except the historical range. In addition, Bray-Curtis dissimilarities overall increased with distance between sites. Relative quantification of ecological processes showed that, for females and males, respectively, drift and dispersal limitation were the primary drivers of community assembly. Collectively, our findings highlight how microbiota structural variance discriminates the western-expanded populations of A. americanum ticks from the Historical range. Spatial autocorrelation, and particularly the detection of non-selective ecological processes, are indicative of geographic isolation. We also found that prevalence of Ehrlichia chaffeensis, E. ewingii, and Anaplasma phagocytophilum ranged from 3.40-5.11% and did not significantly differ by region. Rickettsia rickettsii was absent from our samples. Our conclusions demonstrate the value of synergistic analysis of biogeographic and microbial ecology data in investigating range expansion in A. americanum and potentially other tick vectors as well.

Ivermectin as a novel malaria control tool: Getting ahead of the resistance curse.

Reduction in malaria clinical cases is strongly dependent on the ability to prevent Anopheles infectious bites. Vector control strategies using long-lasting insecticidal nets and indoor residual spraying with insecticides have contributed to significantly reduce the incidence of malaria in many endemic countries, especially in the Sub-Saharan region. However, global progress in reducing malaria cases has plateaued since 2015 mostly due to the increased insecticide resistance and behavioral changes in Anopheles vectors. Additional control strategies are thus required to further reduce the burden of malaria and contain the spread of resistant and invasive Anopheles vectors. The use of endectocides such as ivermectin as an additional malaria control tool is now receiving increased attention, driven by its different mode of action compared to insecticides used so far and its excellent safety record for humans. In this opinion article, we discuss the advantages and disadvantages of using ivermectin for malaria control with a focus on the risk of selecting ivermectin resistance in malaria vectors. We also highlight the importance of understanding how ivermectin resistance could develop in mosquitoes and what its underlying mechanisms and associated molecular markers are, and propose a research agenda to manage this phenomenon.

Do parapatric populations of the ticks Amblyomma tonelliae Nava, Beati & Labruna, 2014 and Amblyomma sculptum Berlese, 1888 (Acari: Ixodidae) hybridize?

This work aimed to determine if the tick species, Amblyomma sculptum and Amblyomma tonelliae, hybridize along their contact zones in Argentina. Free-living adults and nymphs of A. sculptum and A. tonelliae were collected in seven sampling locations of northern Argentina. In four of them, the two species occur in parapatry (possible hybrid zone) whereas in the other three sites, only one species is known to occur. A total of 65 A. sculptum and 65 A. tonelliae from both, allopatric and parapatric populations, were analysed. The nuclear (ITS2) and mitochondrial (COI and 12SrDNA) gene sequences of each tick were amplified and analysed to verify whether or not they could reveal the presence of hybrids among the parapatric samples. No morphological and molecular evidence was found to support the hypothesis of ongoing natural hybridization. Intrinsic postzygotic barriers may be the cause of lack of gene flow between the two species in areas of co-ocurrence. The results can be explained by the length of time the two lineages spent in allopatry since the middle of the Miocene and before their respective distribution range expanded again reaching a narrow secondary contact zone.

Integrative Taxonomy of Dermacentor variabilis (Ixodida: Ixodidae) with Description of a New Species, Dermacentor similis n. sp.

Dermacentor variabilis is the most widely distributed three-host tick in North America, and transmits a variety of pathogens. Within the United States, this species has a discontinuous distribution, widespread east of the Rocky Mountains and with a few populations west of the Rockies. Phylogenetic evidence based on individual markers or relatively small data sets has suggested that populations at both sides of this geographic barrier may correspond to two different species. In this study, we further explore this hypothesis using an integrative taxonomy framework. Both molecular (mitochondrial and nuclear markers) and morphological analyses of specimens collected from central-eastern and western states were performed to explore species delimitation in this taxon. Results from these analyses were consistent, and provide strong evidence that D. variabilis actually corresponds to two species. Herein, the western populations are described as a new species, Dermacentor similis n. sp. The usefulness of integrative taxonomy in the context of species delimitation is also discussed.

Integrating population genetic structure, microbiome, and pathogens presence data in Dermacentor variabilis.

Tick-borne diseases (TBDs) continue to emerge and re-emerge in several regions of the world, highlighting the need for novel and effective control strategies. The development of effective strategies requires a better understanding of TBDs ecology, and given the complexity of these systems, interdisciplinary approaches are required. In recent years, the microbiome of vectors has received much attention, mainly because associations between native microbes and pathogens may provide a new promising path towards the disruption of pathogen transmission. However, we still do not fully understand how host genetics and environmental factors interact to shape the microbiome of organisms, or how pathogenic microorganisms affect the microbiome and vice versa. The integration of different lines of evidence may be the key to improve our understanding of TBDs ecology. In that context, we generated microbiome and pathogen presence data for Dermacentor variabilis, and integrated those data sets with population genetic data, and metadata for the same individual tick specimens. Clustering and multivariate statistical methods were used to combine, analyze, and visualize data sets. Interpretation of the results is challenging, likely due to the low levels of genetic diversity and the high abundance of a few taxa in the microbiome. Francisella was dominant in almost all ticks, regardless of geography or sex. Nevertheless, our results showed that, overall, ticks from different geographic regions differ in their microbiome composition. Additionally, DNA of Rickettsia rhipicephali, R. montanensis, R. bellii, and Anaplasma spp., was detected in D. variabilis specimens. This is the first study that successfully generated microbiome, population genetics, and pathogen presence data from the same individual ticks, and that attempted to combine the different lines of evidence. The approaches and pre-processing steps used can be applied to a variety of taxa, and help better understand ecological processes in biological systems.

Population genetic structure and demographic history of the lone star tick, Amblyomma americanum (Ixodida: Ixodidae): New evidence supporting old records.

Range expansions are a potential outcome of changes in habitat suitability, which commonly result as a consequence of climate change. Hypotheses on such changes in the geographic distribution of a certain species can be evaluated using population genetic structure and demography. In this study we explore the population genetic structure, genetic variability, demographic history of, and habitat suitability for Amblyomma americanum, a North American tick species that is a known vector of several pathogenic microorganisms. We used a double digestion restriction site-associated DNA sequencing technique (dd-RAD seq) and discovered 8,181 independent single nucleotide polymorphisms (SNPs) in 189 ticks from across the geographic range of the species. Genetic diversity was low, particularly when considering the broad geographic range of this species. The edge populations were less diverse than populations belonging to the historic range, possibly indicative of a range expansion, but this hypothesis was not statistically supported by a test based on genetic data. Nonetheless, moderate levels of population structure and substructure were detected between geographic regions. For New England, demographic and species distribution models support a scenario where A. americanum was present in more northern locations in the past, underwent a bottleneck, and subsequently recovered. These results are consistent with a hypothesis that this species is re-establishing in this area, rather than one focused on range expansion from the south. This hypothesis is consistent with old records describing the presence of A. americanum in the northeastern US in the early colonial period.

Borrelia burgdorferi sensu lato infecting Ixodes auritulus ticks in Uruguay.

In the southern cone of South America different haplotypes of Borrelia burgdorferi sensu lato (Bbsl) have been detected in Ixodes spp. from Argentina, southern Brazil, Chile, and Uruguay. So far, Lyme borreliosis has not been diagnosed in Uruguay and the medical relevance of the genus Ixodes in South America is uncertain. However, the growing number of new genospecies of Bbsl in the southern cone region and the scarce information about its pathogenicity, reservoirs and vectors, highlights the importance of further studies about spirochetes present in Uruguay and the region. The aim of this study was to determine the presence of Bbsl in Ixodes auritulus ticks collected from birds and vegetation in two localities of southeastern Uruguay. In total 306 I. auritulus were collected from 392 passerine birds sampled and 1110 ticks were collected by flagging in vegetation. Nymphs and females were analyzed for Borrelia spp. by PCR targeting the flagellin (fla) gene and the rrfA-rrlB intergenic spacer region (IGS). The phylogenetic analysis of Borrelia spp. positive samples from passerine birds and vegetation revealed the presence of four fla haplotypes that form a clade within the Bbsl complex. They were closely related to isolates of Borrelia sp. detected in I. auritulus from Argentina and Canada.

The Amblyomma maculatum Koch, 1844 (Acari: Ixodidae) group of ticks: phenotypic plasticity or incipient speciation?

BACKGROUND: The goal of this study was to reassess the taxonomic status of A. maculatum, A. triste and A. tigrinum by phylogenetic analysis of five molecular markers [four mitochondrial: 12S rDNA, 16S rDNA, the control region (DL) and cytochrome c oxidase 1 (cox1), and one nuclear: ribosomal intergenic transcribed spacer 2 (ITS2)]. In addition, the phenotypic diversity of adult ticks identified as A. maculatum and A. triste from geographically distinct populations was thoroughly re-examined. RESULTS: Microscopic examination identified four putative morphotypes distinguishable by disjunct geographical ranges, but very scant fixed characters. Analysis of the separated mitochondrial datasets mostly resulted in conflicting tree topologies. Nuclear gene sequences were almost identical throughout the geographical ranges of the two species, suggesting a very recent, almost explosive radiation of the terminal operational taxonomic units. Analysis of concatenated molecular datasets was more informative and indicated that, although genetically very close to the A. maculatum - A. triste lineage, A. tigrinum was a monophyletic separate entity. Within the A. maculatum - A. triste cluster, three main clades were supported. The two morphotypes, corresponding to the western North American and eastern North American populations, consistently grouped in a single monophyletic clade with many shared mitochondrial sequences among ticks of the two areas. Ticks from the two remaining morphotypes, south-eastern South America and Peruvian, corresponded to two distinct clades. CONCLUSIONS: Given the paucity of morphological characters, the minimal genetic distance separating morphotypes, and more importantly the fact that two morphotypes are genetically indistinguishable, our data suggest that A. maculatum and A. triste should be synonymized and that morphological differences merely reflect very recent local adaptation to distinct environments in taxa that might be undergoing the first steps of speciation but have yet to complete lineage sorting. Nonetheless, future investigations using more sensitive nuclear markers and/or crossbreeding experiments might reveal the occurrence of very rapid speciation events in this group of taxa. Tentative node dating revealed that the A. tigrinum and A. maculatum - A. triste clades split about 2 Mya, while the A. maculatum - A.triste cluster radiated no earlier than 700,000 years ago.

The microbiome of Haemaphysalis lemuris (Acari: Ixodidae), a possible vector of pathogens of endangered lemur species in Madagascar.

Lemurs are primate species that are endemic to Madagascar. At present, about 90% of lemur species are endangered, and 5 species are among the 25 most endangered primates worldwide. Health status is a major factor impacting the viability of wild populations of many endangered species including lemurs. Given this context, we analyzed the microbiome of 24 specimens of Haemaphysalis lemuris, the most common tick parasitizing lemurs in their native habitats. Ticks were collected from 6 lemur species and microbiomes analyzed using next-generation sequencing. Our results show that the H. lemuris microbiome is highly diverse, including over 500 taxa, 267 of which were identified to genus level. Analysis of the microbiome also shows that there is a distinct "host" (lemur species) component when explaining the differences among and between microbial communities of H. lemuris. This "host" component seems to overwhelm any "locality" (geographic origin of the sample) component. In addition to the microbiome data, targeted PCR was used to test for the presence of three pathogens recently detected in the blood of wild lemurs: Borrelia sp., Candidatus Neoehrlichia sp., and Babesia sp. Overall, the presence of DNA of Rickettsia spp., Bartonella spp., Francisella spp., and a Babesia sp., in H. lemuris, is consistent with the hypothesis that these ectoparasites may act as vector for these pathogens. Further studies assessing vector competence are needed to confirm this hypothesis.

Amblyomma parvum Aragão, 1908 (Acari: Ixodidae): Phylogeography and systematic considerations.

The geographical distribution of Amblyomma parvum Aragão 1908 in the New World is disjunct, with two main clusters separated from each other by the Amazon basin. The main objectives of this study were to further investigate the systematic relationships within A. parvum, to determine whether or not populations from different geographical areas might represent cryptic species, and to reconstruct the phylogeographical evolutionary history of the species. The genetic diversity of A. parvum collected throughout its distributional range was analyzed by using 6 molecular markers: 5 mitochondrial [the small and the large ribosomal subunits 12rDNA and 16SrDNA, the cytochrome oxidase I (COI) and II (COII) and the control region or d-loop (DL)], and one nuclear (ITS2, Inter transcribed spacer 2). Phylogenetic trees were inferred by using maximum parsimony and Bayesian analyses. In addition, node dating was attempted for the main lineages identified phylogenetically. Although mitochondrial and nuclear topologies were not totally congruent, they all identified at least two main supported clusters, a Central American lineage, and a Brazilian-Argentinian lineage. Clade support and divergence values strongly suggest that the two lineages correspond to different taxonomic entities. Node dating placed the split between the Central American and the Brazilian-Argentinian lineages at approximately 5.8-4.9 Mya, just after the progressive replacement of the dry areas that occupied the northern part of South America by the Amazon Basin in the early-mid Miocene. This event might be the cause of fragmentation and putative speciation within the ancestral relatively xerophilic A. parvum population.

Seroepidemiological survey of Rickettsia spp. in dogs from the endemic area of Rickettsia parkeri rickettsiosis in Uruguay.

Rickettsia parkeri rickettsiosis is a vector-borne zoonosis that occurs in some countries of the American continent. Following the first description and determination of the pathogenicity to humans in 2004 in USA, this bacterium has been reported in several South American countries. Human cases have been diagnosed in both Uruguay and Argentina in the past years. This study consisted in a serosurvey of 1000 domestic dogs living in the endemic area of rickettsiosis in Uruguay, where Amblyomma triste is the tick vector. Sera were analyzed by Indirect Immunofluorescence Assay (IFA), against antigens of three different rickettsial species: R. rhipicephali, R. felis and R. parkeri. It was determined that 20.3% of the dogs had antibodies that reacted to at least one of the three species tested, taking as cut off ≥64 titers. Furthermore, 140 of the seropositive dogs (14%) had a titer at least 4 times higher to R. parkeri than those of any of the other species, thus, it was considered that the immune response was stimulated by that species in particular. This is the first serological survey in primary hosts for adults of A. triste in Uruguay, and therefore the first prevalence values are reported. Adult A. triste ticks collected from the environment as well as from dogs were analyzed by PCR in order to confirm the current circulation of the agent in the area. In this matter, two out of 28 ticks from dogs, and 3 out of 53 ticks from the environment were positive, and the corresponding sequence analysis revealed 100% similarity with R. parkeri strain maculatum.

First molecular detection of Rickettsia parkeri in Amblyomma tigrinum and Amblyomma dubitatum ticks from Uruguay.

Rickettsia parkei is the etiological agent of spotted fever in Uruguay, where is transmitted to humans by the tick Amblyomma triste. In the present study, ticks were collected from capybaras (Hydrochoerus hydrochaeris) and domestic dogs during 2011-2012 in different parts of Uruguay. Three out of 11 (27.3%) Amblyomma dubitatum ticks collected from capybaras, and 4 out of 6 (66.7%) Amblyomma tigrinum ticks collected from dogs were shown by molecular analyses to be infected by Rickettsia parkeri strain Maculatum 20. Until the present work, A. triste was the only tick species that was found infected by R. parkeri in Uruguay. This is the first report of R. parkeri infecting these two tick species in Uruguay, expanding the current distribution of this rickettsial pathogen in the country.

A new species of Ornithodoros (Acari: Argasidae), parasite of Microlophus spp. (Reptilia: Tropiduridae) from northern Chile.

A new species, Ornithodoros microlophi (Acari: Argasidae), belonging to the subgenus Alectorobius is described from larvae collected on the lizards Microlophus atacamensis (Donoso-Barros, 1966) and Microlophus quadrivittatus (Tschudi, 1845) (Squamata: Tropiduridae) in continental and insular localities from northern Chile. Larvae of O. microlophi can be distinguished from other Neotropical species of the genus Ornithodoros by a combination of the following characters, namely 10 pairs of ventral setae, venter with 6 pairs of sternal setae, dorsal plate pyriform, 19-21 pairs of dorsal setae (typically 20), 13 pairs are dorsolateral and 7 pairs are central, and hypostome with dental formula 4/4 in medial portion and apex pointed. Phylogenetic analysis of 16S rDNA sequences suggests that O. microlophi represents an independent lineage within Neotropical species of the Argasidae.