Public Engagement with Biotechnology Inside and Outside the Classroom: Community-Focused Approaches.

Biotechnology offers vast benefits to the environment, animals, and human health, and contributes to improving socioeconomic conditions for the public. However, biotechnology innovations continue to trigger public concern and opposition over their potential social, health, and ecological risks. There is an opportunity to increase knowledge and acceptance of biotechnology through engagement, education, and community participation. In this perspective, we highlight crucial factors that shape the public perception of biotechnology and present opportunities for scientists to effectively communicate their ideas while engaging with local and global communities. Initiatives that seek to involve communities in design, development, and adoption processes are crucial for the successful implementation of biotechnology-based solutions.

Intellectual property in entomology: Analysis and perspective on recent trends in global patent publications.

Intellectual property (IP) is an important consideration for entomological research and provides a means to capture value from new discoveries. Herein, we describe an analysis of more than 26 000 patent publications from 2007-2017 related to the field of entomology. These patents were divided among 8000 patent assignees; however, only 5% of the assignees had ≥10 patents. Corporations accounted for the largest share of patents (59%), with individuals (20%), academic institutions (17%) and government organizations (4%) making up the remaining segments. From 2007-2017 the number of entomological patents increased by 400%, with the largest number being from China. However, unlike patents from Europe, Japan or the US, which target a range of countries, the Chinese patents almost exclusively focus on China. Among the array of subjects covered are transgenic insects and plants, repellents, recombinant insect cells, with the highest proportion of patents focused on insecticides (39%), followed by insecticide mixtures (27%) and formulations (21%). The top 30 patent assignees included companies/institutions from China (18), Europe (3), Japan (6) and the US (3). Among the top 12 entities, IP from the US assignees was distributed across insecticides, mixtures and insecticidal traits while those from China were more focused on mixtures. However, given expanding IP numbers from China it is expected that in the future there will be a greater impact on new insecticides and related technologies. © 2020 Society of Chemical Industry.

RNAi targeting of rootworm Troponin I transcripts confers root protection in maize.

Western corn rootworm, Diabrotica virgifera virgifera, is the major agronomically important pest of maize in the US Corn Belt. To augment the repertoire of the available dsRNA-based traits that control rootworm, we explored a potentially haplolethal gene target, wings up A (wupA), which encodes Troponin I. Troponin I, a component of the Troponin-Tropomyosin complex, is an inhibitory protein involved in muscle contraction. In situ hybridization showed that feeding on wupA-targeted dsRNAs caused systemic transcript knockdown in D. v. virgifera larvae. The knockdown of wupA transcript, and by extension Troponin I protein, led to deterioration of the striated banding pattern in larval body muscle and decreased muscle integrity. Additionally, the loss of function of the circular muscles surrounding the alimentary system led to significant accumulation of food material in the hind gut, which is consistent with a loss of peristaltic motion of the alimentary canal. In this study, we demonstrate that wupA dsRNA is lethal in D. v. virgifera larvae when fed via artificial diet, with growth inhibition of up to 50% within two days of application. Further, wupA hairpins can be stably expressed and detected in maize. Maize expressing wupA hairpins exhibit robust root protection in greenhouse bioassays, with several maize transgene integration events showing root protection equivalent to commercial insecticidal protein-expressing maize.

Use of chromatin remodeling ATPases as RNAi targets for parental control of western corn rootworm (Diabrotica virgifera virgifera) and Neotropical brown stink bug (Euschistus heros).

RNA interference (RNAi) is a gene silencing mechanism that is present in animals and plants and is triggered by double stranded RNA (dsRNA) or small interfering RNA (siRNA), depending on the organism. In the western corn rootworm (WCR), Diabrotica virgifera virgifera LeConte (Coleoptera: Chrysomelidae), RNAi can be achieved by feeding rootworms dsRNA added to artificial diet or plant tissues transformed to express dsRNA. The effect of RNAi depends on the targeted gene function and can range from an absence of phenotypic response to readily apparent responses, including lethality. Furthermore, RNAi can directly affect individuals that consume dsRNA or the effect may be transferred to the next generation. Our previous work described the potential use of genes involved in embryonic development as a parental RNAi technology for the control of WCR. In this study, we describe the use of chromatin-remodeling ATPases as target genes to achieve parental gene silencing in two insect pests, a coleopteran, WCR, and a hemipteran, the Neotropical brown stink bug, Euschistus heros Fabricius (Hemiptera: Pentatomidae). Our results show that dsRNA targeting chromatin-remodeling ATPase transcripts, brahma, mi-2, and iswi strongly reduced the fecundity of the exposed females in both insect species. Additionally, knockdown of chd1 reduced the fecundity of E. heros.

The evolving puzzle of autosomal versus Y-linked male determination in Musca domestica.

Sex determination is one of the most rapidly evolving developmental pathways, but the factors responsible for this fast evolution are not well resolved. The house fly, Musca domestica, is an ideal model for studying sex determination because house fly sex determination is polygenic and varies considerably between populations. Male house flies possess a male-determining locus, the M factor, which can be located on the Y or X chromosome or any of the five autosomes. There can be a single M or multiple M factors present in an individual male, in heterozygous or homozygous condition. Males with multiple copies of M skew the sex ratio toward the production of males. Potentially in response to these male-biased sex ratios, an allele of the gene transformer, Md-tra(D), promotes female development in the presence of one or multiple M factors. There have been many studies to determine the linkage and frequency of these male determining factors and the frequency of Md-tra(D) chromosomes in populations from around the world. This review provides a summary of the information available to date regarding the patterns of distribution of autosomal, X-linked and Y-linked M factors, the relative frequencies of the linkage of M, the changes in frequencies found in field populations, and the fitness of males with autosomal M factors vs. Y-linked M. We evaluate this natural variation in the house fly sex determination pathway in light of models of the evolution of sex determination.

A quantitative analysis of a modified feeding method for rearing Cimex lectularius (Hemiptera: Cimicidae) in the laboratory.

BACKGROUND: A modified artificial feeding system was developed for rearing Cimex lectularius and compared with the most widely used artificial feeding method which requires custom-made glassware and a circulating water bath to warm the blood. The petri dish method reduces some of the drawbacks from the current water bath method, such as the possibility of flooding bed bug rearing jars with water or blood and the need for expensive custom-made glassware. RESULTS: Field and lab strain first instar nymphs were given a 15 or 30 min interval to feed for 6-9 weeks. Analysis of bed bugs that fed during a 6 week time period showed that there was no significant difference in the numbers that fed using the petri dish method compared with those that fed using the water bath method. Development of the nymphs also showed that there was no significant difference in the time required to produce adults by either method. CONCLUSION: The petri dish method is an attractive alternative to the water bath method. Set-up is less complex, quicker, multiple jars of bed bugs can be fed at the same time, petri dishes of blood are disposable for easy clean-up and the potential of fatality due to flooding with blood or water is reduced.

Consumption of a durable termite bait matrix by subterranean termites (Isoptera: Rhinotermitidae) and resulting insecticidal activity.

BACKGROUND: A novel durable termite bait was developed to enable continuous bait availability and lengthen the monitoring interval to 1 year. Laboratory studies were conducted to determine the palatability and insecticidal activity of this bait to Reticulitermes flavipes (Kollar), R. virginicus (Banks), R. hesperus Banks, Coptotermes formosanus Shiraki and Heterotermes aureus (Synder). RESULTS: Consumption of the blank durable bait matrix was significantly higher than consumption of a blank preferred textured cellulose matrix (PTC) by R. virginicus, R. flavipes and C. formosanus. R. flavipes, R. hesperus and H. aureus consumed significantly more durable bait than PTC when both contained the active ingredient noviflumuron. All bait treatments resulted in significant mortality relative to the untreated controls. Survivorship of R. virginicus, C. formosanus and H. aureus was 2% or less and not significantly different between the durable bait and PTC treatments containing noviflumuron. The durable bait matrix lagged behind the PTC matrix in mortality over time for all species tested except H. aureus. CONCLUSION: The durable bait was highly palatable and effective in inducing mortality to R. flavipes, R. virginicus, R. hesperus, C. formosanus and H. aureus in the laboratory. This unique bait matrix will be available to termites continuously and allows for an annual monitoring interval. The durability of this bait matrix is unprecedented, allowing for bait to remain active for years and thus providing continuous structural protection.

Selective advantage for IIIM males over YM males in cage competition, mating competition, and pupal emergence in Musca domestica L. (Diptera: Muscidae).

In the house fly, Musca domestica L. (Diptera: Muscidae), sex is usually determined by a dominant factor, M, located on the Y chromosome. However, there are autosomal male (A(M)) populations in which the M factor is located on one or more of the five autosomes (I-V), most commonly on the third chromosome. Herein we report the use of isogenic strains to determine the relative fitness of Y(M) versus III(M) males in three different experiments. First, cages were started with 50% Y(M) and 50% III(M) males, and the frequencies of Y(M) and III(M) males were evaluated across generations. Second, mating competition studies were preformed with these isogenic strains. Third, the relative emergence rates of III(M) versus Y(M) male pupae held at three temperatures for 3 d were examined. All three studies indicate that III(M) males have a greater fitness than Y(M) males. In the cage competition studies, >90% of the males were III(M) after seven generations. III(M) males were more likely to mate than Y(M) males, and a higher percent of III(M) males emerged after being held as pupae at 4, 16, or 28 degrees C for 3 d. The implications of these studies to the distribution of III(M) and Y(M) males in field populations are discussed.

Comparison of two acetylcholinesterase gene cDNAs of the lesser mealworm, Alphitobius diaperinus, in insecticide susceptible and resistant strains.

Two cDNAs encoding different acetylcholinesterase (AChE) genes (AdAce1 and AdAce2) were sequenced and analyzed from the lesser mealworm, Alphitobius diaperinus. Both AdAce1 and AdAce2 were highly similar (95 and 93% amino acid identity, respectively) with the Ace genes of Tribolium castaneum. Both AdAce1 and AdAce2 have the conserved residues characteristic of AChE (catalytic triad, intra-disulfide bonds, and so on). Partial cDNA sequences of the Alphitobius Ace genes were compared between two tetrachlorvinphos resistant (Kennebec and Waycross) and one susceptible strain of beetles. Several single nucleotide polymorphisms (SNPs) were detected, but only one non-synonymous mutation was found (A271S in AdAce2). No SNPs were exclusively found in the resistant strains, the A271S mutation does not correspond to any mutations previously reported to alter sensitivity of AChE to organophosphates or carbamates, and the A271S was found only as a heterozygote in one individual from one of the resistant A. diaperinus strains. This suggests that tetrachlorvinphos resistance in the Kennebec and Waycross strains of A. diaperinus is not due to mutations in either AChE gene. The sequences of AdAce1 and AdAce2 provide new information about the evolution of these important genes in insects.

Changes in the frequency of YM versus IIIM in the housefly, Musca domestica L., under field and laboratory conditions.

In the housefly, Musca domestica L., sex is usually determined by a dominant factor, M, located on the Y chromosome. However, there are 'autosomal male' (AM) populations in which the M factor is located on one or more of the five autosomes (I-V) or on X. We examined changes in the frequency of AM and YM males in North Carolina populations of houseflies after 4 years in the laboratory (NC Lab 02:06) and after 4 or 5 years in the field (NC 2006 and NC 2007). In 2002, 77.7% of the male houseflies were III/III;XYM, 20% were IIIM/III;XX, and 2.3% were IIIM/III;XYM. After 4 years in the laboratory, IIIM/III males disappeared and 17.4% of the males were XMYM. Conversely, 4 years later, the field population was relatively unchanged from 2002. Thus, there was a strong selection against IIIM/III males in the laboratory, but not in the field. Field-collected flies from 2007 indicated a slight increase in the frequency of XYM males and a slight decrease in the frequency of IIIM/III males (relative to 2002 and 2006), suggesting that the relative frequency of XYM and IIIM/III can vary slightly over time in field populations. The detection of XMYM males in 2007 offered the opportunity to evaluate the frequency of the female-determining FD factor, which was found to be present in both the laboratory and field populations, but frequencies varied greatly. The present study represents the first report of FD in houseflies from North America. The significance of these results, relative to observed clines in AM versus YM males, is discussed.

Dynamics of insecticide resistance alleles in house fly populations from New York and Florida.

The frequency of insecticide-resistance alleles for two genes (Vssc1 and CYP6D1) was studied in field collected populations of house flies from two different climates. While the frequency of these resistance alleles in flies at dairies from four states has recently been reported, there is no information on the relative change of these allele frequencies over time. House flies were collected during the 2003-2004 season from New York and Florida before the first application of permethrin, during the middle of the field season, after the final application, and again the following spring (following months without permethrin use). Bioassay results indicated that homozygous susceptible and extremely resistant flies were rare, while moderately and highly resistant individuals were relatively common at all times in both states. The frequency of resistance alleles at the New York dairy rose during the season and declined over the winter, suggesting an overwintering fitness cost associated with these alleles. The super-kdr allele was detected for the first time in North America at the end of 2003. In Florida the frequency of the resistance alleles did not increase during the spray season or decrease during the winter, suggesting there is substantial immigration of susceptible alleles to the Florida dairy and no overwintering fitness cost associated with resistance alleles in this climate. Resistance to permethrin correlated well with the frequency of the Vssc1 and CYP6D1 resistance alleles in flies from New York, but not as well in the population from Florida. This suggests there may be a new resistance mechanism or allele evolving in Florida.

Resistance to cyfluthrin and tetrachlorvinphos in the lesser mealworm, Alphitobius diaperinus, collected from the eastern United States.

The lesser mealworm, Alphitobius diaperinus (Panzer), is an important pest in poultry facilities. The toxicity of cyfluthrin and tetrachlorvinphos to five strains of the lesser mealworm was compared with the toxicity to a susceptible laboratory strain. Bioassays were carried out with both larvae and adults. For the susceptible strain, cyfluthrin and tetrachlorvinphos had similar toxicity to adults, but cyfluthrin was 5 times more toxic to larvae when compared with tetrachlorvinphos. High levels of resistance to tetrachlorvinphos in two beetle strains were detected in both larvae and adults, although these strains were heterogeneous and still contained susceptible individuals. Resistance to cyfluthrin ranged from 1.7- to 9.5-fold for adults and from 0.5- to 29-fold for larvae at the LC(95). Overall, the patterns of resistance did not mirror the insecticide use patterns reported at these facilities. The implications of these results to management of the lesser mealworms are discussed.

A cline in frequency of autosomal males is not associated with insecticide resistance in house fly (Diptera: Muscidae).

Geographic variation in the chromosomal location of the male sex determining factor (M) was studied in four house fly, Musca domestica L., populations from the eastern United States. We found a strong clinal trend (29 degrees 41' latitude in Florida to 44 degrees 2' in Maine) in which the percentage of standard XY(M) males increased with increasing latitude. In Florida, 100% of the males possessed the M factor on the third autosome (III(M)). North Carolina had 20% III(M) males and 2.35% with both Y(M) and III(M). Fewer III(M) males were found in New York (4.35%). Populations from Maine contained 100% XY(M) males. In two of three standard laboratory-susceptible strains, all males carried M on an autosome ("autosomal males" or A(M)): CS (III(M)) and SRS (V(M)). Insecticide bioassays of four field-collected strains led us to conclude that resistance is not correlated with sex determination over a broad range of insecticides. For example, high levels of resistance to permethrin (86-99% survival at a diagnostic concentration) were found in all four field-collected strains. The five other insecticides evaluated showed varying levels of resistance among field strains. We conclude that a cline is present in house fly populations from the eastern United States with 100% III(M) males in the south and entirely Y(M) males in the north and that insecticide resistance is not a key factor influencing the evolution or linkage of M.