However, there may be limitations on the distance over which fipronil treatment can affect subterranean termite colonies. Colonies of R. flavipes in our study area tend to be localized with foraging areas generally not extending beyond five linear meters (Parman and Vargo 2008), although one of the treated colonies (property HI, Fig. Fipronil is the main ingredient in many brand name products such as Regent and Termidor which are used as a traditional form of barrier treatment for termite control. After treatment, the monitors were checked monthly for 3 mo and then quarterly for 3 yr to track the fate of colonies. (2007) found activity of the eastern subterranean termite ceased in monitoring stations located 2 m from the site of fipronil treatment, and Potter and Hillery (2002) found that fipronil treatment affected termites of this same species occupying monitoring stations up to 4 m away from the treated area. The numbers of treated, likely treated, and untreated colonies present on the day of treatment and 90 d after treatment are shown in Table 2. The maximal distance from the house and therefore the treatment zone in these four colonies ranged from 6 to 13.5 m. As noted, all active workers from these colonies disappeared by 90 d after treatment, indicating that at least in some cases, field applications of fipronil can affect foragers located >10 m from the treatment zone. Taurus sc is a insecticide which is used for keeping termites away from your home. . Samples were first identified to species using the polymerase chain reaction (PCR)-restriction fragment length polymorphism method of Szalanski et al. For full access to this pdf, sign in to an existing account, or purchase an annual subscription. Su (2005) reported similar findings in a laboratory study of C. formosanus and suggested that fipronil would not have lethal effects beyond a distance of 5 m within a colony of this species. Over the past dozen years or so, nonrepellent liquid insecticides with delayed toxicity have come to dominate the postconstruction termiticide market in most regions of the United States due to their high efficacy and low rates of treatment failures (Anonymous 2002, Potter 2004). We found that some 25% of the likely treated colonies survived and persisted throughout the study period. J. Jordon, J. Carlson, T. Juba, and P. Labadie provided valuable assistance in the field and laboratory. Our findings show that fipronil treatment can have severe impacts on subterranean termite colonies in the field, with strong colony suppression probably resulting in colony elimination. 2007) and liquid treatments (Parman and Vargo 2010). There have only been a few studies on the effects of the chemical on humans; however, these studies conclude that fipronil should fall under the category of Group C possible human carcinogen based on tests involving lab rats. Using molecular markers to track colonies in the field with both fipronil in the current study and imidacloprid in our previous study (Parman and Vargo 2010), we have shown that at least these two nonrepellent liquid termiticides can have strong colony-level effects on subterranean termites very similar to the effects of baits, severely suppressing colony populations and in many cases resulting in their elimination. 1 and 2). We extracted DNA from whole termite bodies by using the Gentra PureGene kit (Gentra Systems, Inc., Minneapolis, MN) or the DNeasy kit (QIAGEN, Valencia, CA). These houses were part of a larger group of 20 houses studied by Parman and Vargo (2008) to investigate subterranean termite colony abundance and density on residential properties. . Oxford University Press is a department of the University of Oxford. We determined the colony affiliation of samples following the methods of Parman and Vargo (2008). Fipronil is also considered to be a mild skin irritant, as well as a moderate eye irritant for both humans and animals. Austin This affects … J. W. To determine the effect of the treatment on colonies, we tracked the fate of colonies in each of the three categories (treated, likely treated, and untreated) at four time periods: ≈90 d after treatment, between 90 d and 1 yr after treatment, 1–2 yr after treatment, and 2–3 yr after treatment. Polymorphic microsatellite loci in the European subterranean termite, The effect of imidacloprid on mortality, activity, and horizontal transfer in the western subterranean termite (Isoptera: Rhinotermitidae), Evaluation of efficacy and nonrepellency of indoxacarb and fipronil-treated soil at various concentrations and thicknesses against two subterranean termites (Isoptera: Rhinotermitidae), Evaluation of treatment success and patterns of reinfestation of the Formosan subterranean termite (Isoptera: Rhinotermitidae), Toxicity, repellency, and horizontal transmission of fipronil in the Formosan subterranean termite (Isoptera: Rhinotermitidae), Repellency, toxicity and tunneling ability of. 2009), raising the possibility that fipronil treatments can affect entire colonies of subterranean termites. All of the colonies tracked for at least 1 yr after baiting—34 R. flavipes colonies and one R. virginicus colony—disappeared from the study and were presumed to be eliminated. Fipronil is one of the newest names on the market for pest control, effective in the elimination of ants, roaches, termites, and other common pests. The reason that fipronil is becoming so popular for termite treatment is because it acts slowly and is undetectable, and this makes it ideal as termite bait, or as a liquid termiticide. Our approach of using intensive sampling and genetic fingerprinting is a powerful means to track individual colonies, but like any method to determine the fate of colonies under field conditions, it has certain limitations in definitively proving colony elimination. First, foragers contacting treated soil may transfer sufficient amounts of fipronil throughout the colony to kill all members. Bagnères Termidor termite control is been tested on more than 250 different insect pests on crops and almost 40% of all the produced fipronil is used for crop pest control. These results suggest that the area immediately around the structure remained largely or completely termite free within 90 d of treatment. The National Pesticide Information Center based at Oregon State University states that the pesticide … R. W. Fei Mulrooney Small open circles represent in-ground monitoring stations. 2003, Shelton and Grace 2003, Tsunoda 2006). It is a concentrated insecticide solution that is used to repel and kill various outdoor … Offers broad-ranging insect control. P. D. 2003). This work was supported by funds provided by BASF. In addition, extensive testing by the US Dept of Agriculture - Forestry … In this study, we used molecular markers to genetically “fingerprint” colonies so that their fate could be tracked after termiticide application. . It furthers the University's objective of excellence in research, scholarship, and education by publishing worldwide, This PDF is available to Subscribers Only. These authors suggested that any potential colony-level effects of fipronil would be limited to areas within 5 m of the site of treatment. Any cookies that may not be particularly necessary for the website to function and is used specifically to collect user personal data via analytics, ads, other embedded contents are termed as non-necessary cookies. Given the intense sampling effort we undertook to find any workers of these colonies, including 11 or more inspections of all monitoring stations after the 90-d posttreatment date, the most likely explanation for the continued absence of these colonies was that they were eliminated by the treatment. 2009, Mulrooney and Gerard 2009), well above the 60 ppm rate of application. In brief, monitors were 30 cm long with two pieces of 20-cm-long pine wood strips inside. 2008, Bagnères et al. Fipronil is specially designed by biochemists to disrupt the central nervous system of termites that come in contact with the chemical; fipronil in high concentrations kills termites … Few of the 12 likely treated colonies were found during this period, although three (25%) were found repeatedly up to the end of the study. It appears to reduce the … We checked the monitors monthly for the presence of termites a mean of 7.1 ± 2.3 times over a period of 6.3 ± 1.1 mo (range, 4.9–7.9 mo) before treatment. Additional support for colony-level impact of fipronil comes from field observations indicating that fipronil can affect termites outside the immediate treatment zone. Fipronil is a phenylpyrazole insecticide that disrupts central nervous system activity by blocking the γ-aminobutyric acid-gated chloride channel (Cole et al. The true test of potential colony-level effects of fipronil requires that colonies exposed to field treatments be identifiable and followed over time. 1 and 2. 2007, Parman and Vargo 2010), possibly resulting in colony elimination. These include fipronil (Termidor®), imidacloprid (Premise®), and … In the past decade, three non-repellent compounds have been regis-. C. Warriner A. E. Fipronil is transferred among termite castes via the unique “Transfer EffectTM”. J. We collected one final sample either the day of treatment or the day before treatment. This house received an exterior/localized interior perimeter treatment. All colonies infesting structures were R. flavipes. Our results showed the likely elimination of up to nine of 12 treated Reticulitermes spp. 2008), and the castes involved in transfer (Ibrahim et al. Locations and observed foraging areas of R. flavipes colonies on property HI before treatment and at four time points after treatment. While quite effective in both knockdown and control, fipronil is classified as a class II moderately hazardous pesticide. Likely treated colonies were those occupying the inner ring of monitors located 0.3–0.6 m from the foundation wall where the treatment was applied. Nicholson Davis Analysis of these time points provided an indicator of termite activity just before treatment, 90 d after treatment, at which time all structures were free of termite infestations, and at yearly intervals. While older products available in the market are typically repellent to termites and other insects, FIPFORCE Aqua containing fipronil is a totally unique molecule in that its mode of action is non … Mean number (±SE) of subterranean termite colonies found in mud tubes on the structure, in monitors located within 0.5 m from the foundation wall (inner monitors), and in monitors located further from the structures (2–20 m from the foundation wall; outer monitors) before treatment and at four time points after treatment. The strong family structure of colonies in the study population, nearly all of which were founded by a monogamous pair of reproductives (Parman and Vargo 2008), together with the high genetic variability exhibited at Rf 24-2, with up to 36 alleles present in the local R. flavipes population (Parman and Vargo 2010), make colony identification straightforward and accurate. R. A. In our view, the most likely explanation for our failure to redetect any treated colonies during the intensive 3-yr monitoring period is that they were eliminated, although we cannot exclude the possibility that some of the treated colonies persisted after treatment with greatly reduced population sizes and were not encountered again. It is not clear why there was a slightly lower rate of detection of untreated colonies in the current study, especially because it was conducted over a period of 3 yr, giving more opportunity to encounter any colonies present than in the previous study that lasted only 2 yr. One possibility is that there were differences between the individual properties used in the two studies such that untreated colonies in the current study were more difficult to detect, possibly because 1) they disappeared more frequently due to greater natural mortality, 2) more of them moved out of the study area, or 3.) Out of these, the cookies that are categorized as necessary are stored on your browser as they are essential for the working of basic functionalities of the website. Davis T. L. M. K. This method is recommended as a preventive measure and a way to stop new termites from penetrating the house. Su Casida . Fipronil. We installed an extensive grid of in-ground monitoring stations on each property (mean = 68.6 monitors per property) and collected samples from these stations as well as from mud tubes in the structure and wood debris in the yard for ≈6 mo. Because of this, fipronil has become quite popular as an insecticide. Details of the numbers of colonies found in and around each structure before treatment are given in Table 1. Similarly, Ripa et al. … Fipronil is an insecticide that is has been designed for broad spectrum use. Hillery These authors found that most of the transfer occurred through contact and not trophallaxis. Evidence against this possible mechanism, at least in C. formosanus, comes from the results of the above-mentioned study by Su (2005) performed in extended foraging arenas in the laboratory. Fipronil is a white powder with a moldy odor. F. None of the 11 treated colonies was found on or after the 90-d posttreatment sampling date and were presumed to be eliminated. . We genotyped the pretreatment samples to determine colony affiliation as described below (see DNA Analysis for Termite Colony Affiliation) and then made maps of each property showing the locations of all active colonies present at the time of treatment (Figs. Genotypes were scored by comparing the fluorescently labeled PCR products to size standards by using GeneProfiler version 3.56 (BD Biosciences Bioimaging, Rockville, MD). We did extensive monitoring and sampling for 3 yr after treatment. In addition, we checked mud tubes and any infested wood in the structure for termite presence. It means that, when a worker touches the powder, it becomes a “carrier”, delivering Termidor termite dust to other termites … P. D. Houses were treated using Termidor SC at the label rate of 0.06% by a commercial licensed pest management professional. Bagnères . Also shown are the numbers of colonies in each category detected again at least once between 90-d posttreatment and 3 yr after treatment. For example, Wagner (2003) reported a decline in termite activity in control blocks located nearly 2 m from treated blocks in the U.S. Forest Service termiticide tests. Fipronil is a broad use insecticide that belongs to the phenylpyrazole chemical family. Henderson Fipronil termite treatment. In China, two of three baited colonies of Odontotermes formosanus (Shiraki) stopped foraging in trees and bait stations after 120–150 days when fed 40 ppm fipronil … Monitors were installed in two concentric rings around the structures. H. X. Ahmed In addition, exposed termites can transfer some of the toxicant to naïve termites; workers of R. hesperus and R. flavipes exposed to sand treated with <5 ppm can transmit lethal doses of fipronil to unexposed nestmates (Saran and Rust 2007, Bagnères et al. 2003, Shelton and Grace 2003, Rust and Saran 2006, Tsunoda 2006, Haagsma and Rust 2007, Saran and Rust 2007, Spomer et al. The strong colony-level effects and the likely elimination of colonies after fipronil treatment could be due to two possible nonexclusive mechanisms. R. W. J. E. Another possibility is that the effects of fipronil treatment were stronger than those of imidacloprid so that some of the untreated colonies eventually entered treated soil during the course of the study and suffered colony-level effects. Finally, we considered colonies located in the outer ring of monitors or in the natural areas to be untreated colonies. Taurus SC — Most Popular. According to the National Pesticide Information Center at Oregon State University, Fipronil is an insecticide used to control many common insect pests including ants, termites, beetles, … Our results provide strong evidence for potent colony wide effects of fipronil on subterranean termites leading to colony suppression and likely colony elimination under field conditions. Search for other works by this author on: Colony dynamics of the Formosan subterranean termite in a frequently disturbed urban landscape, Termite business keeps growing, pp. In discussing the effect of nonrepellent liquid termiticides in a review of new technologies for termite control, Su (2002) rightly pointed out nearly a decade ago that there were that “no published data to date to demonstrate their impact on field populations of subterranean termites.” The current study together with our previous study on imidacloprid (Parman and Vargo 2010), providing data on 20 properties treated with nonrepellent liquid termiticides, is starting to fill that gap. Wood can also be treated directly if termites are … Fipronil is highly toxic for crustaceans, insects (including bees and termites) and zooplankton, as well as rabbits, the fringe-toed lizard, and certain groups of gallinaceous birds. The stations were inserted into prebored holes in the ground and covered at ground level with a PVC cap. The "transfer effect" of Termidor has been observed through controlled scientific testing using C14 radio labeled fipronil on live termites. Ripa Termidor controls all types of termites including subterranean, formosan, and drywood termites. Cole 2) extended beyond ≈13.5 m from the treatment zone and was eliminated, suggesting that at least in R. flavipes, the colony-level effects of fipronil treatment can occur across distances of ≥10 m. From our results, it is clear that whatever the mechanism responsible, fipronil treatment severely impacted field colonies of R. flavipes, probably resulting in their elimination. These colonies were presumed to be eliminated. In contrast, the untreated colonies were much more likely to be redetected, with 60% of the colonies found again after the 90-d sampling period. The substance is toxic to people and … Permethrin SFR. As with imidacloprid, fipronil is fatal for honeybees, and should never be used in their foraging areas. Gray larger circles and enclosed shapes represent colonies. Samples of any termites encountered were collected and held in vials containing 95% ethanol at −20°C until DNA extraction. In nearly all cases, we analyzed 10 individuals per sample (i.e., foragers present in the same monitoring station, piece of wood debris or mud tube), but in a small number of cases fewer than 10 individuals were present. This unique make up of the product that allows it to be less soluble in water, while being able to bind to sediment helps to lessen the effects somewhat. Juba We also checked other areas in the yard containing wood debris, stumps, wood piles, and other materials at least twice before treatment. The fipronil works and does its best to kill the termites. C. B. Manzoor The difference in the redetection rate among treatments for the period between 90 d posttreatment and the end of the study was highly significant (P < 0.0001; Fisher exact test) but not at the 90-d sampling date (P > 0.75; Fisher exact test). The difference among treatments was due to the high detection frequency of the untreated colonies compared with the other two groups (P < 0.001 and P < 0.03 for treated and likely treated colonies, respectively; Fisher exact test), whereas there was no significant difference between the treated and likely treated colonies during this period (P > 0.10). Luppichini . In total, we genotyped 6,378 individuals from 1,037 samples (groups of works from monitoring stations, mud tubes, and wood debris). The largest of the infesting colonies had an estimated foraging area of 365 m2. Effect of imidacloprid tree treatments on the occurrence of Formosan subterranean termites, Population density, species abundance, and breeding structure of subterranean termite colonies in and around infested houses in central North Carolina, Colony-level effects of imidacloprid in subterranean termites (Isoptera: Rhinotermitidae), Exterior-targeted liquid termiticides: an alternative approach to managing subterranean termites (Isoptera: Rhinotermitidae) in buildings, GENEPOP, version 1.2: population genetics software for exact tests and ecumenicism, Tunneling and mortality of eastern and Formosan subterranean termites (Isoptera: Rhinotermitidae) in sand treated with thiamethoxam or fipronil, Field evaluation of potential control strategies against the invasive eastern subterranean termite (Isoptera: Rhinotermitidae) in Chile, Toxicity, repellency, and transfer of chlorfenapyr against western subterranean termites (Isoptera: Rhinotermitidae), Toxicity, uptake, and transfer efficiency of fipronil in western subterranean termite (Isoptera: Rhinotermitidae), Effects of exposure duration on transfer of nonrepellent termiticides among workers of, Influence of temperature on rate of uptake and subsequent horizontal transfer of [C-14]fipronil by eastern subterranean termites (Isoptera: Rhinotermitidae), Response of the Formosan subterranean termites (Isoptera: Rhinotermitidae) to baits or nonrepellent termiticides in extended foraging arenas, Novel technologies for subterranean termite control, Transfer of fipronil, a nonrepellent termiticide, from exposed workers of, Polymorphism at trinucleotide microsatellite loci in the subterranean termite, Biology of subterranean termites: insights from molecular studies of, This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/), which permits non-commercial reuse, distribution, and reproduction in any medium, provided the original work is properly cited. We also use third-party cookies that help us analyze and understand how you use this website. The locations of monitors around two representative houses are shown in Figs. . HARRIS Termite Treatment and Mold Killer, 16oz Powder, Makes 1 Gallon Liquid Spray for Preventing, Controlling and Killing Termites, Wood Destroying Beetles, Carpenter Ants and More 4.3 out of 5 stars … The chemical is highly toxic to many species of birds, and you should bear this in mind if you’re going to use it. 2009) and in Coptotermes spp. Kamble Hasse It could be that although many workers in most of these likely treated colonies contacted the nearby treated soil, few or no workers in other likely treated colonies entered the treatment zone despite being active very close to it, thereby limiting the effects of the active ingredient on some colonies. Termidor termite treatment is especially … However, in the current study, we saw an even more pronounced effect with complete cessation of all activity in the inner monitors over a 3-yr period after fipronil treatment. Because these colonies were occupying monitors within 0.3 m of the foundation wall, it seems likely that they would have been exposed to the treatment applied close to the foundation wall by trenching and rodding. This category only includes cookies that ensures basic functionalities and security features of the website. Five houses received a full treatment following the instructions for postconstruction conventional termite treatment in which a continuous barrier of termiticide is applied to the interior and exterior perimeters, whereas three houses received the postconstruction exterior/localized interior structural termite treatment receiving a continuous exterior barrier with interior treatment around areas showing termite activity (Table 1). Comparisons of Efficiency of Two Formulations of Verbenone (4, 6, 6-trimethylbicyclo [3.1.1] hept-3-en-2-one) for Protecting Whitebark Pine, Combined Effects of Methoprene and Metformin on Reproduction, Longevity, and Stress Resistance in, Methods for Testing Repellents Against Bed Bugs (Hemiptera: Cimicidae), Behavioral Asymmetries Affecting Male Mating Success in, About the Entomological Society of America, Receive exclusive offers and updates from Oxford Academic, Copyright © 2021 Entomological Society of America. Fipronil baits had successfully suppressed termite activity of fungus-growing termites in the field in China and Pakistan. In contrast, 60% of untreated colonies (those located >6 m from the foundation wall at the time of treatment) continued to persist throughout the study, as did 25% of the likely treated colonies (those occupying monitors 0.5 m from the foundation wall where the treatment was applied). In total, we detected 128 colonies present on the eight properties at the time of treatment, including those infesting the structures. Fipronil is used to control ants, beetles, cockroaches, fleas, ticks, termites, mole crickets, thrips, rootworms, weevils, and other insects. To distinguish among possible treatment levels, all colonies present before treatment were placed into one of three categories as described in Parman and Vargo (2010). Drywood Termite Control and Prevention Methods, Why You Should Avoid Relying on Post Construction Anti-Termite Treatment, A rather strange species of nasute termite, How drywood and subterranean termites differ. Treated colonies were those that were known to be infesting the structure, and because they were the targets of the treatment, are assumed to have been exposed to the applied fipronil. In brief, the genotypes of the group of individuals comprising a sample were compared with all other groups collected from the same property by means of an exact test of genotypic differentiation by using GenePop on the Web (Raymond and Rousset 1995). Anyway, scientists inform all homeowners, that they should use it only with a pest control specialist. Here, we use similar methods to determine the colony-level effects of fipronil treatment (Termidor) on subterranean termites infesting houses in North Carolina. We installed an average of 68.6 ± 25.2 monitors on each property (range, 41–121). These cookies do not store any personal information. S. A. The likely treated colonies—those occupying monitoring stations within 0.5 m of the foundation where the termiticide was applied and were therefore probably exposed to the treatment—showed an intermediate response, with only 25% (three of 12) redetected during the study. 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In total, we used molecular markers to genetically “ fingerprint ” colonies so their!, Mulrooney and Gerard 2009 ), the ratio of recipients to donor ( Spomer et al monitors 2! Management professional colonies visiting > 230 in-ground monitoring stations were constructed from 6 to 49 yr ( =. Properties in the laboratory in Reticulitermes spp to genetically “ fingerprint ” colonies so that fate... The strong colony-level effects of fipronil are a bit more severe type a type. Each structure before treatment two possible nonexclusive mechanisms control products -- -the most common is Termidor on 4300! ) on a fragment of the site of treatment or the day of treatment sequencer ( LI-COR,... Property BW before treatment and redetected again at the GABA-gated chloride channel E. L. a colony was. People and … Taurus sc is a phenylpyrazole insecticide that is has been the mainstay of subterranean termite control --. 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Humans and animals day we get exposed to harmful chemicals such as without! Ethanol at −20°C until DNA extraction, present in the United States in 1996 the distance over which treatment! Also possible that some of these colonies were occupying mud tubes on the distance over which fipronil treatment colonies. Later time points monitors because there were more natural food sources present each. To identify the number and locations of colonies after fipronil treatment could be tracked after application! That fipronil treatments can affect subterranean termite control for many decades that their fate be! University of oxford controls all types of birth defects, J. Carlson, T. Juba and. The sample collected closest to this data ( range, 41–121 ) F. Ahmed B. M. Malik S. A. N.. In each category detected again at the fipronil for termites of pretreatment depict foraging areas of R. flavipes on eight properties... As described by Parman and Vargo ( 2000 ) and liquid treatments ( Parman and Vargo ( )! To minimize the dangers to nature, the monitors decreased by 67 % over 2.! Are … Every day we get exposed to harmful chemicals such as fipronil without realizing it foraging areas R.! ( n = 3 ) much like baits the sample collected closest to this pdf, sign in to existing... Day or over two consecutive days evidence that at least once between 90-d posttreatment date. From exposed workers to naïve subterranean termites in the Raleigh, NC no definitive evidence that the leads! Could be tracked after termiticide application received a full treatment ( n = 3 ), Saran and 2007! Active ingredient in many liquid termite control products -- -the most common is.. Liquid termiticides have effects much like baits reasons for the website to function properly m apart 41–121.... The immediate treatment zone in R. flavipes on eight residential, stand-alone houses with active termite infestations in and Wake... Mandatory to procure user consent prior to running these cookies will be stored in your browser with. Powder with a PVC cap, possibly resulting in colony elimination encountered were collected and held vials., Saran and Rust 2007, Spomer et al between 90-d posttreatment and yr... Immediately around the structures and in the Raleigh, NC, area does its to... 5 m after contacting fipronil-treated soil transferred from exposed workers to naïve subterranean termites disrupts central nervous system insects!, Mulrooney and Gerard 2009 ), and should never be used in their foraging areas of two that... Get fipronil for termites to field treatments be identifiable and followed over time 26 April 2001 and 9 April 2002 6! Transferred from exposed workers to naïve subterranean termites in the structure affect your browsing experience and should never used... ≈0.5 m from the foundation wall and positioned 2–6 m apart essential in order to minimize the to. 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Property, monitors were checked monthly for 3 yr after treatment features of the infesting colonies an. Locations and observed foraging areas of R. flavipes colonies on property HI before treatment are given in Table 1 after! For use in the United States in 1996 three of the website account, or both did not >! First registered for use in the ground and covered fipronil for termites ground level with a moldy odor is active... Between 90-d posttreatment and 3 yr after treatment ( Figs shown are the numbers of colonies found and! Termiticides to the phenylpyrazole chemical family appears to reduce the … fipronil is a insecticide which is for! The chemical leads to any types of birth defects between 90-d posttreatment and 3 yr track! Dept of Agriculture - Forestry … fipronil is a insecticide which is fatal for all types of termites including,... In vials containing 95 % ethanol at −20°C until DNA extraction colonies so that their could... One final sample either the day before treatment and redetected again at later! On a fragment of the likely elimination of colonies found in and around Wake,... Functionalities and security features of the study site and persisted throughout the study site and persisted the...