Impact of involving the community in entomological surveillance of Triatoma infestans (Klug, 1834) (Hemiptera, Triatominae) vectorial control.

BACKGROUND: Vectorial transmission is the principal path of infection by Trypanosoma cruzi, the parasite that causes Chagas disease. In Argentina, Triatoma infestans is the principal vector; therefore, vector control is the main strategy for the prevention of this illness. The Provincial Program of Chagas La Rioja (PPCHLR) carries out entomological evaluation of domiciliary units (DUs) and spraying of those where T. infestans is found. The lack of government funds has led to low visitation frequency by the PPCHLR, especially in areas with a low infestation rate, which are not prioritized. Therefore, seeking possible alternatives to complement control activities is necessary. Involving householders in entomological evaluation could be a control alternative. The major objective was to determine the cost of entomological evaluation with and without community participation. METHODS: For entomological evaluation without community participation, PPCHLR data collected in February 2017 over 359 DUs of the Castro Barros Department (CBD) were used. For entomological evaluation with community participation, 434 DUs of the same department were selected in November 2017. Each householder was trained in collecting insects, which were kept in labeled plastic bags, recovered after 2 weeks, and analyzed in the laboratory for the presence of T. cruzi. Using householders' collection data, a spatial scan statistic was used to detect clusters of different T. infestans infestations. Entomological evaluation costs with and without community participation related to the numbers of DUs visited, DUs evaluated, and DUs sprayed were calculated and compared between methodologies. In addition, the number of DUs evaluated of the DUs visited was compared. RESULTS: According to the results, the triatomines did not show evidence of T. cruzi infection. Spatial analysis detected heterogeneity of T. infestans infestation in the area. Costs related to the DUs visited, evaluated, and sprayed were lower with community participation (p < 0.05). In addition, more DUs were evaluated in relation to those visited and a greater surface area was covered with community participation. CONCLUSION: Participation of the community in the infestation survey is an efficient complement to vertical control, allowing the spraying to be focused on infested houses and thus reducing the PPCHLR's costs and intervention times.

A Simple, Inexpensive Method for Mark-Recapture of Ixodid Ticks.

Mark-recapture techniques have been widely used and specialized to study organisms throughout the field of biology. To mark-recapture ticks (Ixodida), we have created a simple method to mark ticks using nail polish applied with an insect pin secured in a pencil that allows for a variety of questions to be answered. For measuring tick control efficacy, estimating population estimates, or measuring movement of ticks, this inexpensive mark-recapture method has been easily applied in the field and in the lab to provide useful data to answer a variety of questions about ticks.

A new cost-effective and fast direct PCR protocol for insects based on PBS buffer.

Insect DNA barcoding is a species identification technique used in biodiversity assessment and ecological studies. However, DNA extraction can result in the loss of up to 70% of DNA. Recent research has reported that direct PCR can overcome this issue. However, the success rates could still be improved, and tissues used for direct PCR could not be reused for further genetic studies. Here, we developed a direct PCR workflow that incorporates a 2-min sample preparation in PBS-buffer step for fast and effective universal insect species identification. The developed protocol achieved 100% success rates for amplification in six orders: Mantodea, Phasmatodea, Neuroptera, Odonata, Blattodea and Orthoptera. High and moderate success rates were obtained for five other species: Lepidoptera (97.3%), Coleoptera (93.8%), Diptera (90.5%), Hemiptera (81.8%) and Hymenoptera (75.0%). High-quality sequencing data were also obtained from these amplifiable products, allowing confidence in species identification. The method was sensitive down to 1/4th of a 1-mm fragment of leg or body and its success rates with oven-dried, ethanol-preserved, food, bat guano and museum specimens were 100%, 98.6%, 90.0%, 84.0% and 30.0%, respectively. In addition, the pre-PCR solution (PBS with insect tissues) could be used for further DNA extraction if needed. The workflow will be beneficial in the fields of insect taxonomy and ecological studies due to its low cost, simplicity and applicability to highly degraded specimens.

Sequencing historical specimens: successful preparation of small specimens with low amounts of degraded DNA.

Despite advances that allow DNA sequencing of old museum specimens, sequencing small-bodied, historical specimens can be challenging and unreliable as many contain only small amounts of fragmented DNA. Dependable methods to sequence such specimens are especially critical if the specimens are unique. We attempt to sequence small-bodied (3-6 mm) historical specimens (including nomenclatural types) of beetles that have been housed, dried, in museums for 58-159 years, and for which few or no suitable replacement specimens exist. To better understand ideal approaches of sample preparation and produce preparation guidelines, we compared different library preparation protocols using low amounts of input DNA (1-10 ng). We also explored low-cost optimizations designed to improve library preparation efficiency and sequencing success of historical specimens with minimal DNA, such as enzymatic repair of DNA. We report successful sample preparation and sequencing for all historical specimens despite our low-input DNA approach. We provide a list of guidelines related to DNA repair, bead handling, reducing adapter dimers and library amplification. We present these guidelines to facilitate more economical use of valuable DNA and enable more consistent results in projects that aim to sequence challenging, irreplaceable historical specimens.

The Technical and Performance Characteristics of a Low-Cost, Simply Constructed, Black Light Moth Trap.

The universal mercury vapor black light trap is an effective device used for collecting moth specimens in a wide variety of habitats; yet, they can present challenges for researchers. The mercury vapor trap is often powered by a heavy automotive battery making it difficult to conduct extensive surveys in remote regions. The mercury vapor trap also carries a considerable financial cost per trap unit, making trapping challenging with low research budgets. Here, we describe the development and trapping properties of a lighter, simply constructed, and less expensive trap. The LED funnel trap consists of a funnel, soda bottles with plastic vanes, and is powered by rechargeable 9-V batteries. Two strips of low-wavelength LEDs are used as attractants. We tested the trapping parameters of this trap design compared to a standard mercury vapor trap over 10 trap nights in a suburban woodlot in the summer of 2015. The mercury vapor trap caught significantly more moth individuals than the LED trap (average of 78 vs 40 moths per trap night; P < 0.05), and significantly more species than the LED trap (23 vs 15 per trap night; P < 0.05); the mercury vapor trap caught a total of 104 macromoth species over the duration of the study, compared to a total of 87 by the LED trap. Despite the lower yields, the low cost of the LED trap (<$30 ea.) makes it superior to the mercury vapor trap in cost-acquisition per moth species and per moth individual trapped. The LED trap may be a viable alternative to the standard mercury vapor trap, facilitating insect trapping in more diverse settings.

Microsatellite loci cross-species transferability in Aedes fluviatilis (Diptera:Culicidae): a cost-effective approach for population genetics studies.

BACKGROUND: Aedes fluviatilis is a neotropical mosquito species thought to be a potential vector of Yellow Fever viruses and can be infected with Plasmodium gallinaceum in laboratory. A better understanding of its genetic structure is very important to understand its epidemiologic potential and how it is responding to urbanization. The objective of this study was to survey the transferability of microsatellites loci developed for other Aedes to Ae. fluviatilis. FINDINGS: We tested in Ae. fluviatilis 40 pairs of primers known to flank microsatellite regions in Aedes aegypti, Aedes albopictus and Aedes caspius, and found eight loci that amplified consistently. The number of alleles per locus ranged from 2 to 15, and the expected heterozygosity ranged from 0.09 to 0.85. CONCLUSIONS: We found that several microsatellite primers successfully transferred to Ae. fluviatilis. This finding opens avenues for cost-effective optimization of high-resolution population genetic tools.

A cost-effective, community-based, mosquito-trapping scheme that captures spatial and temporal heterogeneities of malaria transmission in rural Zambia.

BACKGROUND: Monitoring mosquito population dynamics is essential to guide selection and evaluation of malaria vector control interventions but is typically implemented by mobile, centrally-managed teams who can only visit a limited number of locations frequently enough to capture longitudinal trends. Community-based (CB) mosquito trapping schemes for parallel, continuous monitoring of multiple locations are therefore required that are practical, affordable, effective, and reliable. METHODS: A CB surveillance scheme, with a monthly sampling and reporting cycle for capturing malaria vectors, using Centers for Disease Control and Prevention light traps (LT) and Ifakara Tent Traps (ITT), were conducted by trained community health workers (CHW) in 14 clusters of households immediately surrounding health facilities in rural south-east Zambia. At the end of the study, a controlled quality assurance (QA) survey was conducted by a centrally supervised expert team using human landing catch (HLC), LT and ITT to evaluate accuracy of the CB trapping data. Active surveillance of malaria parasite infection rates amongst humans was conducted by CHWs in the same clusters to determine the epidemiological relevance of these CB entomological surveys. RESULTS: CB-LT and CB-ITT exhibited relative sampling efficiencies of 50 and 7%, respectively, compared with QA surveys using the same traps. However, cost per sampling night was lowest for CB-LT ($13.6), followed closely by CB-ITT ($18.0), both of which were far less expensive than any QA survey (HLC: $138, LT: $289, ITT: $269). Cost per specimen of Anopheles funestus captured was lowest for CB-LT ($5.3), followed by potentially hazardous QA-HLC ($10.5) and then CB-ITT ($28.0), all of which were far more cost-effective than QA-LT ($141) and QA-ITT ($168). Time-trends of malaria diagnostic positivity (DP) followed those of An. funestus density with a one-month lag and the wide range of mean DP across clusters was closely associated with mean densities of An. funestus caught by CB-LT (P < 0.001). CONCLUSIONS: CB trapping schemes appear to be far more affordable, epidemiologically relevant and cost-effective than centrally supervised trapping schemes and may well be applicable to enhance intervention trials and even enable routine programmatic monitoring of vector population dynamics on unprecedented national scales.

Regional insect inventories require long time, extensive spatial sampling and good will.

Understanding how faunistic knowledge develops is of paramount importance to correctly evaluate completeness of insect inventories and to plan future research at regional scale, yet this is an unexplored issue. Aim of this paper was to investigate the processes that lead to a complete species inventory at a regional level for a beetle family. The tenebionid beetles of Latium region (Italy) were analysed as a case study representative of general situations. A comprehensive faunistic database including 3,561 records spanning from 1871 to 2010 was realized examining 25,349 museum specimens and published data. Accumulation curves and non-parametric estimators of species richness were applied to model increase in faunistic knowledge over time, through space and by collectors' number. Long time, large spatial extent and contribution of many collectors were needed to obtain a reliable species inventory. Massive sampling was not effective in recovering more species. Amateur naturalists (here called parafaunists) were more efficient collectors than professional entomologists. Museum materials collected by parafaunists over long periods and large spatial extent resulted to be a cost effective source of faunistic information with small number of collected individuals. It is therefore important to valuate and facilitate the work of parafaunists as already suggested for parataxonomists. By contrast, massive collections by standardized techniques for ecological research seem to be of scarce utility in improving faunistic knowledge, but their value for faunistic studies may be enhanced if they are conducted in poorly surveyed areas.

Occurrence and identification of risk areas of Ixodes ricinus-borne pathogens: a cost-effectiveness analysis in north-eastern Italy.

BACKGROUND: Ixodes ricinus, a competent vector of several pathogens, is the tick species most frequently reported to bite humans in Europe. The majority of human cases of Lyme borreliosis (LB) and tick-borne encephalitis (TBE) occur in the north-eastern region of Italy. The aims of this study were to detect the occurrence of endemic and emergent pathogens in north-eastern Italy using adult tick screening, and to identify areas at risk of pathogen transmission. Based on our results, different strategies for tick collection and pathogen screening and their relative costs were evaluated and discussed. METHODS: From 2006 to 2008 adult ticks were collected in 31 sites and molecularly screened for the detection of pathogens previously reported in the same area (i.e., LB agents, TBE virus, Anaplasma phagocytophilum, Rickettsia spp., Babesia spp., "Candidatus Neoehrlichia mikurensis"). Based on the results of this survey, three sampling strategies were evaluated a-posteriori, and the impact of each strategy on the final results and the overall cost reductions were analyzed. The strategies were as follows: tick collection throughout the year and testing of female ticks only (strategy A); collection from April to June and testing of all adult ticks (strategy B); collection from April to June and testing of female ticks only (strategy C). RESULTS: Eleven pathogens were detected in 77 out of 193 ticks collected in 14 sites. The most common microorganisms detected were Borrelia burgdorferi sensu lato (17.6%), Rickettsia helvetica (13.1%), and "Ca. N. mikurensis" (10.5%). Within the B. burgdorferi complex, four genotypes (i.e., B. valaisiana, B. garinii, B. afzelii, and B. burgdorferi sensu stricto) were found. Less prevalent pathogens included R. monacensis (3.7%), TBE virus (2.1%), A. phagocytophilum (1.5%), Bartonella spp. (1%), and Babesia EU1 (0.5%). Co-infections by more than one pathogen were diagnosed in 22% of infected ticks. The prevalences of infection assessed using the three alternative strategies were in accordance with the initial results, with 13, 11, and 10 out of 14 sites showing occurrence of at least one pathogen, respectively. The strategies A, B, and C proposed herein would allow to reduce the original costs of sampling and laboratory analyses by one third, half, and two thirds, respectively. Strategy B was demonstrated to represent the most cost-effective choice, offering a substantial reduction of costs, as well as reliable results. CONCLUSIONS: Monitoring of tick-borne diseases is expensive, particularly in areas where several zoonotic pathogens co-occur. Cost-effectiveness studies can support the choice of the best monitoring strategy, which should take into account the ecology of the area under investigation, as well as the available budget.

Inexpensive trap for monitoring the green June beetle.

Green June beetle, Cotinis nitida (L.), is an important pest of grapes, peaches, blackberries, blueberries, apples, and pears. Currently, there is no inexpensive, commercially available lure or trap that could serve monitoring green June beetle adults. The objective of this study was to develop and optimize an inexpensive bottle trap baited with isopropanol to attract and capture green June beetle adults. Bottle traps baited with 8 mm diameter cotton wicked dispensers emitted from 9 to 43 ml isopropanol in 48 h and maintained that alcohol at a fairly constant concentration compared with the prototypical bottle trap with large surface evaporation of isopropanol poured into the bottom of the trap. Over 5 d, the isopropanol in the wicked dispensers remained at the same stable concentration of 45-44.5%, whereas isopropanol concentration in the bottom of prototypical traps dropped from 45% to approximately 11% after 24 h and to 0.2% by 48 h. Bottle traps with isopropanol dispensers and cotton wicks of 4, 6, or 8 mm in diameter caught significantly more green June beetles than did prototypical bottle traps with no dispensers. Isopropanol concentrations of 45.5, 66, and 91% attracted more green June beetle adults than the lower concentrations. Significantly more green June beetle adults were attracted to traps with dispensers set at 1.3 m height than those at lower heights, and traps topped with a blue, orange, or white band captured more green June beetle adults than those with bands of other colors. The optimized bottle trap is made from recycled transparent polyethylene terephthalate beverage bottle (710-ml; 24 oz.) with a blue, orange, or white band, baited with an 8 mm cotton wick dispenser of 45.5% isopropanol and hung at a height of 1.3 m. Cost and uses for this trap are discussed.

An inexpensive, versatile mosquito rearing chamber for field, laboratory, and classroom investigations.

An inexpensive mosquito rearing chamber for field, laboratory, and classroom investigations is described. The rearing chamber is made from plastics recycled from peanut butter jars and room deodorizers. The top of the chamber requires mesh material and gluing. The cost for the rearing chamber is negligible. The design of the chamber allows for direct field collecting of larvae and for easy knock down/cold storage of emerged adults. In addition to its use in field and laboratory investigations, the chamber is an excellent device for classroom study of insect metamorphosis.

Finding the economics in economic entomology.

To recommend new pest management tactics and strategies to farmers and policy makers, economic entomologists must evaluate the economics of biologically reasonable approaches. We collected data to determine how frequently these economic evaluations occur. We discovered from our survey of entomological journals representing the discipline of economic entomology that < 1% of research papers published since 1972 include economic evaluations of pest management tactics. At least 85% of these analyses were performed by entomologists and not economists. Much of the research on economic evaluations is performed without special funds granted by agencies separate from the authors' institutions. In the United States, USDA competitive grants supported 20% of the economic evaluations published since 2000. However, only approximately 12% of the projects funded since 2000 by three sections of the USDA (Crops at Risk, Risk Avoidance and Mitigation Program, and Pest Management Alternatives Program) resulted in publications concerning economic evaluations. If the purpose of economic entomology is to ultimately determine the value of different kinds of tactics, the discipline may need to take steps to enhance the research that supports these evaluations.

Evaluation of the shaking technique for the economic management of American foulbrood disease of honey bees (Hymenoptera: Apidae).

Shaking is a nonantibiotic management technique for the bacterial disease American foulbrood (AFB) (Paenibacillus larvae sensu Genersch et al.), in which infected nesting comb is destroyed and the adult honey bees, Apis mellifera L. (Hymenoptera: Apidae), are transferred onto uncontaminated nesting material. We hypothesized that colonies shaken onto frames of uninfected drawn comb would have similar reductions in AFB symptoms and bacterial spore loads than those shaken onto frames of foundation, but they would attain higher levels of production. We observed that colonies shaken onto drawn comb, or a combination of foundation and drawn comb, exhibited light transitory AFB infections, whereas colonies shaken onto frames containing only foundation failed to exhibit clinical symptoms. Furthermore, concentrations of P. larvae spores in honey and adult worker bees sampled from colonies shaken onto all comb and foundation treatments declined over time and were undetectable in adult bee samples 3 mo after shaking. In contrast, colonies that were reestablished on the original infected comb remained heavily infected resulting in consistently high levels of spores, and eventually, their death. In a subsequent experiment, production of colonies shaken onto foundation was compared with that of colonies established from package (bulk) bees or that of overwintered colonies. Economic analysis proved shaking to be 24% more profitable than using package bees. These results suggest that shaking bees onto frames of foundation in the spring is a feasible option for managing AFB in commercial beekeeping operations where antibiotic use is undesirable or prohibited.

"The horizon opened up very greatly": Leland O. Howard and the transition to chemical insecticides in the United States, 1894-1927.

The transition to synthetic chemicals as a popular method of insect control in the United States was one of the most critical developments in the history of American agriculture. Historians of agriculture have effectively identified the rise and charted the dominance of early chemical insecticides as they came to define commercial agriculture between the emergence of Paris green in the 1870s and the popularity of DDT in the 1940s and beyond. Less understood, however, are the underlying mechanics of this transition. this article thus takes up the basic question of how farmers and entomologists who were once dedicated to an impressively wide range of insect control options ultimately settled on the promise of a chemically driven approach to managing destructive insects. Central to this investigation is an emphasis on the bureaucratic maneuverings of Leland O. Howard, who headed the Bureau of Entomology from 1894 to 1927. Like most entomologists of his era, Howard was theoretically interested in pursuing a wide variety of control methods--biological, chemical, and cultural included. In the end, however, he employed several tactics to streamline the government's efforts to almost exclusively support arsenic and lead-based chemical insecticides as the most commercially viable form of insect control. While Howard in no way "caused" the national turn to chemicals, this article charts the pivotal role he played in fostering that outcome.

Multiple orifice distribution system for placing green lacewing eggs into verticel larval rearing units.

Green lacewings are widely used biological control agents for various insect pests. To meet the needs of growers, green lacewings are being mass-reared commercially around the world. A common salt shaker has been used regularly to distribute eggs into Verticel lacewing larval rearing units. This technique is time consuming and inefficient because the number of eggs distributed in each cell is inconsistent. The multiple orifice distribution (MOD) system described here greatly improved egg distribution efficiency by increasing the percentage of Verticel cells containing the desired one to four eggs per cell (i.e., 40.8 and 52.1% by using salt shaker method versus 61.9% by using the MOD system). This mechanical system significantly reduced the labor and time involved in the process and would cost under $3500. In addition, this new system could be modified for distribution of other insect eggs.

Field trials of an improved cost-effective device for detecting peridomestic populations of Triatoma infestans (Hemiptera: Reduviidae) in rural Argentina.

An improved device for detecting peridomestic Triatoma infestans consisting of one-liter recycled Tetra Brik milk boxes with a central structure was tested using a matched-pair study design in two rural areas in Argentina. In Olta (La Rioja), the boxes were installed beneath the thatched roofs and on the vertical wooden posts of each peridomestic structure. After a 5-month exposure, at least one of the recovered boxes detected 88% of the 24 T. infestans-positive sites, and 86% of the 7 negative sites by timed manual collections at baseline. In Amamá (Santiago del Estero), the boxes were paired with the best performing prototype tested before (shelter unit). After 3 months, some evidence of infestation was detected in 89% (boxes) and 79% (shelters) of 18-19 sites positive by timed collections, whereas 19% and 16% of 32 negative sites were positive, respectively. Neither device differed significantly in the qualitative or quantitative collection of every sign of infestation. The installation site did not modify significantly the boxes' sampling efficiency in both study areas. As the total cost of each box was half as expensive as each shelter unit, the boxes are thus the most cost-effective and easy-to-use tool for detecting peridomestic T. infestans currently available.