Julieta Brambila

Use of DNA barcodes to identify invasive armyworm Spodoptera species in Florida.

Abstract A critical component for sustaining adequate food production is the protection of local agriculture from invasive pest insects. Essential to this goal is the ability to accurately distinguish foreign from closely related domestic species, a process that has traditionally required identification using diagnostic morphological “keys” that can be both subtle and labor-intensive. This is the case for the Lepidopteran group of insects represented by Spodoptera, a genus of Noctuidae “armyworm” moths that includes several important agricultural pests. Two of the most destructive species, Spodoptera littoralis (Boisduval) (Lepidoptera: Noctuidae) and S. litura (F.) are not yet established in North America. To facilitate the monitoring for these pests, the feasibility of using DNA barcoding methodology for distinguishing between domestic and foreign Spodoptera species was tested. A DNA barcoding database was derived for a subset of Spodoptera species native to Florida, with an emphasis on those attracted to pheromone blends developed for S. litura or S. littoralis. These were then compared to the barcode sequences of S. litura collected from Taiwan and S. littoralis from Portugal. Consistent discrimination of the different species was obtained with phenetic relationships produced that were generally in agreement with phylogenetic studies using morphological characteristics. The data presented here indicate that DNA barcoding has the potential to be an efficient and accurate supplement to morphological methods for the identification of invasive Spodoptera pests in North America.

Monitoring for Exotic Spodoptera Species (Lepidoptera: Noctuidae) in Florida

Abstract Trapping studies were conducted in 2 Florida locations to determine if 3 Old World Spodoptera Guenée species were present. Commercially-produced lures for S. exempta (Walker), S. littoralis (Boisduval), and S. litura (F.), plus a S. litura lure made by the USDA-APHIS-CPHST laboratory at Otis ANGB in Massachusetts, were used with plastic Unitraps and placed near 2 orchid nurseries in Lake and Miami-Dade counties. One S. litura male moth was identified from collections made in Apr 2007; no other exotic species were found in either location. However, thousands of resident species were collected, including S. albula (Walker) (= S. sunia Guenée), S. dolichos (F.), S. eridania (Stoll), S. exigua (Hübner), (J. E. Smith), and S. pulchella (Herrich-Schäffer). This study exposed the amount of labor and level of technical knowledge needed for scientists involved in finding exotic Spodoptera species.

A Major Pest of Cotton, Oxycarenus hyalinipennis (Heteroptera: Oxycarenidae) in the Bahamas

Oxycarenus hyalinipennis (Fig. 1) is a member of the superfamily Lygaeoidea, family Oxycarenidae (Henry 1997), subfamily Oxycareninae. Originally named Aphanus tardus var. hyalinipennis , this species was described from Italy by Costa in 1847. Adults measure 4 to 4.3 mm long and are tapered anteriorly and rounded posteriorly; their thorax, head, antennae, and femora are black and their wings are translucent white, but this species is highly variable (Slater 1972). Males and females have similar coloration, but males are slightly smaller than females. There are 5 instars (Fig. 1), which have pink to red abdomens. Oxycarenus hyalinipennis has numerous synonyms and common names, but as an important pest of cotton worldwide, it is commonly referred to as “the cotton seed bug.” It not only feeds on other plants in the order Malvales, especially in the family Malvaceae, but also in Tiliaceae and Sterculiaceae (Slater & Baranowski 1994). Adults and nymphs suck oil from mature seeds and fluid from leaves of young stems to obtain moisture, according to Ananthakrishnan et al. (1982). If cotton seed bugs are present in sufficient numbers, the cotton fiber becomes stained during processing by the maceration of their bodies. In Israel during outbreaks, this insect has been reported as aggregating on various trees and shrubs including dates, figs, avocados, and persimmons. Many of these fruits were damaged, purportedly due to contamination with a pungent odor (Nakache & Klein 1992). Oxycarenus hyalinipennis has a worldwide distribution, although it has not been reported from North America. Because it has been intercepted numerous times at U.S. ports of entry and because it is already established on islands near the U.S. mainland, its arrival in the U.S. is expected. In the West Indies, Baranowski & Slater (2005) report it from the Turks and Caicos, Bahamas, Cayman Islands, and Hispaniola. It was not found during a preliminary survey along the entire eastern coast of Florida in 2006 (Brambila & Dobbs, unpublished data, 2007). The objectives of this study were to (1) collect observational data on O. hyalinipennis , and (2) determine the infestation levels of this pest on cotton in Great Inagua. Great Inagua was the site of a failed attempt to grow and cultivate cotton, Gossypium hirsutum L., in the early 20 th century. These cotton plantations lasted only a few years, but the cotton itself has persisted in a feral state and can be found as a roadside weed near Matthewtown and former settlements (Fig. 2). This made Great Inagua an excellent location to look for and study infestation levels of Oxycarenus hyalinipennis (Costa), a known pest of cotton. Three locations were chosen for survey along the western coast of Great Inagua (Fig. 2). Only

Combining Tpi and CO1 Genetic Markers to Discriminate Invasive Helicoverpa armigera From Local Helicoverpa zea (Lepidoptera: Noctuidae) Populations in the Southeastern United States

Abstract The recent establishment of the Old World pest Helicoverpa armigera (Hübner) into South America has had significant economic consequences and places the rest of the hemisphere at risk, emphasizing the need for improved methods of monitoring. A major complication is that a sibling species endemic to the New World, Helicoverpa zea (Boddie), is morphologically very similar, with the two species capable of producing fertile hybrids in the laboratory. The consequences of such hybridization in the field are uncertain, but could result in significant and unpredictable changes in the timing, range, and pesticide susceptibilities of Helicoverpa infestations. The objective here is to provide new genetic resources applicable to Helicoverpa populations in northern Florida and neighboring states (a region at risk for H. armigera) that can distinguish the two species and possible hybrids. The genetic variability in segments of the mitochondrial cytochrome oxidase 1 (CO1) and the Z-linked triosephosphate isomerase (Tpi) genes were determined for H. zea from the southeastern United States. These were compared to DNA sequences from H. armigera specimens from Morocco, Australia, and Europe. Phylogenetic network analysis showed a clear demarcation between the two species for all gene segments. These results extend earlier studies establishing CO1 as marker for discriminating the Helicoverpa species complex and introduce a new sex-linked genomic marker. The CO1 and Tpi markers in combination provide a more accurate and sensitive method than existing techniques for identifying hybridization between H. zea and H. armigera and could potentially be used to extrapolate the likely source of invasive H. armigera populations.

Assessing the Usefulness of DNA Barcoding to Identify Oxycarenus hyalinipennis (Hemiptera: Oxycarenidae) in Florida, a Potentially Invasive Pest of Cotton

ABSTRACT Invasive insects present an ongoing challenge to the safety of U.S. agriculture. A current threat to the U.S. cotton industry is Oxycarenus hyalinipennis (Costa), commonly known as the cotton seed bug. Populations are found throughout most of the world except for North America, and the southeastern U.S. is believed to provide a favorable environment for its establishment. A major component in efforts to control the spread of invasive pests is the rapid and accurate identification of intercepted specimens. Unfortunately, O. hyalinipennis belongs to an incompletely characterized taxon where the assignment of species identity by simple morphological keys is often problematic. In this study, we assessed the potential of DNA barcoding to facilitate the identification of the cotton seed bug in field-collected specimens.

PARAGONATAS DIVERGENS (HEMIPTERA: RHYPAROCHROMIDAE): FIRST CONFIRMED RECORD FOR FLORIDA AND THE UNITED STATES

A specimen of Paragonatas divergens (Distant) (Hemiptera: Heteroptera: Rhyparochromidae: Lethaeini) was collected in northeastern Lee County, Florida, May 16, 2003, by the senior author during a routine survey. The specimen was taken by general sweep-net collecting along a roadside at the intersection of North River Road (Hwy. 78) and Parkinson Road. A variety of herbaceous plants at the site was sampled but ragweed, Ambrosia artemisiifolia L., was particularly abundant. In general, members of the Lethaeini are poorly represented in Florida. Slater and Baranowski (1990) noted the presence of only three genera: Cistalia Stal, Cryphula Stal, and Paragonatas Barber, each with a single species reported from Florida. One additional genus, Valtissius Barber, has been reported from Florida as Petissius Distant (Van Duzee 1917) but without specific locality data. Paragonatas divergens occurs throughout tropical America and the West Indies (O’Donnell 1986; Slater 1964; Slater and Baranowski 1990). This species has been referred to as the most common lethaeine in the New World (O’Donnell 1986), yet it was not previously known to occur in the United States. Palmer and Bennett (1988) reported this bug (as Palagonatas [sic]) on Baccharis halimifolia L. in Florida but without a specific locality. The current location of the specimen upon which their record was based is unknown, resulting in Slater and Baranowski (1990) considering this a questionable Florida record. The single female was initially identified by the senior author. The identification was later confirmed by Thomas J. Henry (USDA-ARS-SEL, Washington, D.C.) on July 7, 2003, and the specimen is currently housed at the National Museum of Natural History, Smithsonian Institution, Washington, D.C. Paragonatas contains two known species, P. costaricensis (Distant) and the type species, P. divergens (Distant). The former has been known to occur in the South Florida counties of MiamiDade and Monroe for some time, but P. divergens has not been reliably reported from the United States until now (Slater and Baranowski 1990). Paragonatas divergens , however, has been intercepted on imported commodities numerous times at several of Florida’s ports of entry (Fort Lauderdale, Jacksonville, Miami, and West Palm Beach) as well as other ports of entry in at least seven other states, including Texas and California. The generic limits for Paragonatas need to be redefined. O’Donnell (1986) stated that she could find no common diagnostic characters that united the two species. Slater and Baranowski (1990) questioned whether the two species are congeneric, observing that P. divergens more closely resembles species of Cistalia . In addition, we have noted substantial differences in the general appearance of some P. divergens specimens from the West Indies compared with those from Central America. In our opinion, at least one additional species occurs in the West Indies. The specimen captured in Florida in May, 2003, more closely resembles those we have seen from Central America. Paragonatas divergens is a ground-dwelling species that apparently feeds on fallen seeds of a variety of plants characteristic of old field habitats (Slater and Baranowski 1990). Paragonatas divergens can be easily distinguished from P. costaricensis by the following characters: dorsal surface dull, or at most sub-shining, pubescent, reddishbrown, usually with an obvious comma-shaped, pale macula distally on the corium; forefemora possessing 2-3 acute spines distally; scent gland auricle slender, not strongly curved posteriorly. P. costaricensis : dorsal surface strongly polished, brown, lacking a distinct comma-shaped pale macula distally on the corium; forefemora lacking acute spines distally, scent gland auricle broad and distinctly curved posteriorly. The authors express appreciation to Thomas J. Henry for verifying the identification and for reviewing an earlier version of this manuscript. In addition, we thank Charles F. Brodel (USDAAPHIS-PPQ, Miami, FL) and James A. Slater (University of Connecticut, Storrs, CT) for critically reviewing the manuscript. We thank T. J. Henry and A. G. Wheeler (Clemson University, Clemson, SC) for companionship during the trip in which the specimen was taken.

Efficacies of Four Pheromone-Baited Traps in Capturing Male Helicoverpa (Lepidoptera: Noctuidae) Moths in Northern Florida

Abstract Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) is a serious pest of grain, row, and vegetable crops throughout much of the world, although it is currently not established in the United States. USDA-APHIS and the Cooperative Agricultural Pest Survey program are charged with the responsibility to monitor for this insect pest. The adult stage is the easiest to monitor using pheromone-baited traps. Traps must be easy to handle, portable and cost effective so that they can provide high quality specimens for identifiers. This study was conducted from spring through the fall in 2010 and 2011 to compare the trapping efficacy and cost-effectiveness of 4 pheromone-baited traps for male Helicoverpa moths. Over 11,600 Helicoverpa moths were captured, all identified as the corn earworm, H. zea (Boddie). The Pherocon® 1C “sticky” trap generally captured the fewest number of males, while equal numbers of moths were captured in a wire cone, ScentryTM Heliothis, and Universal (Unitrap) Moth “bucket” trap when moderate populations were present. Wire cone traps performed statistically better when high populations were present. The sticky traps captured the highest number of non-target insects, most being ants, flies, and beetles. Overall, the average corn earworm per trap vs. cost ratio for bucket traps was higher than the other traps, suggesting that more moths per dollar would be captured using these traps.

Efficiency of Trapping Systems for Detecting Tuta absoluta (Lepidoptera: Gelechiidae)

ABSTRACT Tuta absoluta Meyrick (Lepidoptera: Gelechiidae), a pest of tomato, was recently detected in Panama in Central America and now threatens to expand into the important tomato production areas of Mexico and the United States. Moths caught in T. absoluta pheromone-baited traps must be removed and dissected to confirm the species present before containment and mitigation strategies are put in place. Timely processing of traps can be hindered by the presence of numerous similar nontarget moths that cannot be easily prescreened. Trapping systems using dry bucket traps or Delta traps with either hot melt pressure sensitive adhesives (HMPSA) or cool melt adhesives were evaluated for their effectiveness in trapping T. absoluta and for their ease in allowing identification of nontarget moths. Delta traps in Panama with HMPSA and cool melt adhesives both trapped T. absoluta with equal efficacy. In Florida, nontarget moths were easier to prescreen from bucket traps and HMPSA inserts. Importantly, moths found in bucket traps as well as on cool melt adhesive inserts were of a lower quality than those on HMPSA inserts, making identification more difficult. Studies conducted in Florida and Panama tomato and potato fields showed that commercially produced pheromones containing only the main pheromone component ((3E, 8Z, 11Z)-tetradecatrien-1-yl acetate) or containing both the main and minor pheromone component ((3E, 8Z)-tetradecadien-1-yl) attracted nontarget moths. Survey programs, particularly large-scale ones, should consider the application of alternative trapping systems or new adhesives available in order to facilitate the visual prescreening of nontarget moths.

A New Species of the Genus Sinoe (Lepidoptera: Gelechiidae: Litini) from Florida

ABSTRACT A new species of Gelechiidae, Sinoe capsana sp. nov., is reported from southern Florida, USA. All specimens were recently caught in pheromone traps near tomato fields during an early detection survey for the tomato leafminer, Tuta absoluta (Meyrick), a pest native to South America that does not occur in North America. Sinoe capsana sp. nov. is described and imagos and male genitalia are illustrated.

FIRST RECORDS FOR DIEUCHES ARMATIPES (HETEROPTERA: RHYPAROCHROMIDAE) IN NORTH AMERICA

Dieuches armatipes (Walker), a moderately large lygaeoid that occurs throughout Africa, has been discovered recently in several Florida counties. Previously, it had been intercepted by the U.S. Department of Agriculture, APHIS/PPQ, at several Florida ports-of-entry. In Africa, D. armatipes feeds on peanuts ( Arachis hypogaea L.) during the harvesting process; thus, this species has the potential to become a serious pest in northern Florida. In this paper, we give the first United States records of D. armatipes , review the literature, provide a diagnosis, and discuss its pest potential. Henry and Froeschner (1993) gave the first New World records of D. armatipes from the West Indies, based on collections from Dominican Republic, Grand Cayman, Jamaica, and St. Kitts. It also has been collected in St. Croix and Cayman Brac (R. M. Baranowski 2003, pers. comm.). In the Old World, D. armatipes is distributed throughout Africa and as far north as Spain (Andalucia) (Eyles 1973). In his world review, Eyles (1973) redescribed D. armatipes and included a key to all species of Dieuches , photographs of adults, and illustrations of genital capsules, parameres, and spermathecae. At present, 132 species of Dieuches are known (Eyles 1995). Henry and Froeschner (1993) redescribed the adult of D. armatipes and included dorsal and lateral photographs of an adult female to help distinguish it from other rhyparochromid species in the United States.