Laboratory and Field Responses of the Mosquito, Culex quinquefasciatus, to Plant-Derived Culex spp. Oviposition Pheromone and the Oviposition Cue Skatole

Laboratory and field studies were conducted on the oviposition behavior of the pathogen-vectoring mosquito, Culex quinquefasciatus, in response to the oviposition pheromone 6-acetoxy-5-hexadecanolide, produced from a renewable plant resource, Kochia scoparia (Chenopodiaceae) (plant-derived pheromone, PDP), and via an established synthetic route (synthetic oviposition pheromone, SOP). Responses to the oviposition cue skatole (3-methylindole), presented individually and in combination with the plant-derived and synthetic oviposition pheromone, were also studied. Both laboratory and field assays showed that PDP and SOP were equally attractive. Synergistic effects were observed with one combination of PDP and skatole combinations in laboratory assays. Synergy was also observed under field conditions. SOP and skatole combinations showed additive effects in laboratory assays, but were not tested in field bioassays. Although synergism has been previously demonstrated with combinations of SOP and polluted waters, the work presented here is the first example of synergy between a specific oviposition attractant and the oviposition pheromone. Furthermore, the efficacy of mosquito pheromone produced from a cheap, renewable botanical source has been demonstrated.     

Influence of Synthetic Oviposition Pheromone and Volatiles from Soakage Pits and Grass Infusions Upon Oviposition Site-Selection of Culex Mosquitoes in Tanzania

The response of Culex mosquitoes to (5R,6S)-6-acetoxy-5-hexadecanolide (the synthetic oviposition pheromone, SOP), emanations from soakage pit water (SPW), and grass infusions (GI) was studied in pit latrines in Muheza, Tanzania. Water treated with the synthetic oviposition pheromone received more egg rafts of Culex quinquefasciatus Say and Cx. cinereus Theobald than did the untreated water (P < 0.001). The residual activity of SOP did not decrease over a nine-day period, with the geometric mean number of egg rafts laid in the SOP-treated water varying daily between 2.5 and 8.9 as compared to 0.1–1.6 egg rafts laid in tap water. However, SOP did not attract ovipositing mosquitoes to nonbreeding sites, and both treatment and control received few eggs. SPW and GI attracted ovipositing Cx. quinquefasciatus, Cx. cinereus, and Cx. tigripes Grandpré and de Charmoy with the number of eggs rafts deposited in bowls containing SPW or GI being larger than that deposited in bowls with tap water (P < 0.05). More egg rafts of Cx. quinquefasciatus were deposited in SOP-treated water than in SPW (P < 0.05). When SOP was compared with SOP + SPW, more egg rafts of Cx. quinquefasciatus were deposited in bowls with the latter combination than in bowls containing SOP or SPW only (P < 0.05), indicating a synergistic effect between SOP and SPW. Similarly, when SOP-treated water and GI were compared with SOP + GI, more egg rafts of Cx. quinquefasciatus were laid in the latter bowl, indicating a synergistic effect between SOP and GI. The result suggest that the combined use of SOP and organically enriched water can be employed in monitoring of Cx. quinquefasciatus for control programs. This is the first record of the attraction of Cx. cinereus and Cx. tigripes to oviposition stimuli.     

Oviposition Responses of Gravid Female <Emphasis Type="Italic">Culex quinquefasciatus</Emphasis> to Egg Rafts and Low Doses of Oviposition Pheromone Under Semifield Conditions

Semifield experiments were conducted to study the oviposition of gravid Culex quinquefasciatus females in response to one or 10 egg rafts, or 3.0 μg of synthetic oviposition pheromone, (-)-(5R,6S)-6-acetoxy-5-hexadecanolide, a dose equivalent to 10 egg rafts. These treatments were added to the small bowls filled with either hay infusion or water in a small (0.3-m spacing) or a large-square design (3.4-m spacing). Oviposition choice was more pronounced in the “small square” assays. Mean number of egg rafts laid in response to a single egg raft in an oviposition jar filled with hay infusion was significantly greater than with all other treatments. When the oviposition pheromone dose was increased from one to 10 rafts or when 3.0 μg synthetic oviposition pheromone were dispensed on a floating receptacle, synergistic effects were observed between the oviposition pheromone and the hay infusion at both distances. This study is the first demonstration that the amount in a single raft induces oviposition of gravid Cx. quinquefasciatus females under semi-natural conditions.     

Synergism between chemical attractants and visual cues influencing oviposition of the mosquito,Culex quinquefasciatus (Diptera: Culicidae)

Physical factors, such as the color of the oviposition substrate, have seldom been compared with chemical cues in their ability to elicit oviposition behavior in mosquitoes. The role of dyed oviposition waters in attracting ovipositingCulex quinquefasciatus was examined in laboratory experiments. Oviposition waters dyed with ink were found to be significantly attractive to ovipositingCx. quinquefasciatus when compared to distilled water controls. Experiments demonstrated that the mosquitoes were responding to the increased optical density of the dyed oviposition water rather than volatile components of the dye. Ink was also considered in combination with chemical oviposition cues. No comparative data exist on the effect of physical and chemical factors presented together on the oviposition behavior ofCx. quinquefasciatus. Waters dyed with ink acted synergistically with a five-component chemical attractant mixture (3-methylindole, indole, 4-methylphenol, 4-ethylphenol, and phenol) in inducing oviposition in a 2×2 factorial experiment.     

Pheromone chirality of african palm weevil,Rhynchophorus phoenicis (F.) and palmetto weevil,Rhynchophorus cruentatus (F.) (Coleoptera: Curculionidae)

There are four stereoisomers of both 3-methyl-octan-4-ol, the aggregation pheromone of the African palm weevil,Rhynchophorus phoenicis (F.) and 5-methyl-octan-4-ol, the aggregation pheromone of the palmetto weevil,Rhynchophorus cruentatus (F.). Synthetic stereoisomers of 3-methyl-octan-4-ol and 5-methyl-octan-4-ol were baseline-separated on a Cyclodex-B fused silica column. Use of this column in gas chromatographic-electroantennographic detection (GC-EAD) and GC-mass spectrometric (GC-MS) analyses revealed that only one stereoisomer, (3S,4S)-3-methyl-octan-4-ol and (4S,5S)-5-methyl-octan-4-ol, is produced by maleR. phoenicis and maleR. cruentatus, respectively, and elicits good antennal responses by conspecific male and female weevils. In field trapping experiments, withR. phoenicis in Côte d'Ivoire andR. cruentatus in Florida, (3S,4S)-3-methyl-octan-4-ol and (4S,5S)-5-methyl-octan-4-ol strongly enhanced attraction of fresh palm tissue, whereas other stereoisomers were behaviorally benign. Stereoisomeric 3-methyl-octan-4-ol and 5-methyl-octan-4-ol may be utilized to monitor and/or manage populations of these two palm weevils.     

Attractancy and species specificity of 6-acetoxy-5-hexadecanolide,a mosquito oviposition attractant pheromone

Four stereoisomers of 6-acetoxy-5-hexadecanolide, a mosquito oviposition attractant pheromone, were bioassayed for their activity on mosquitoes. Only (–)-(5R,6S) isomer was active in attractingCulex quinquefasciatus Say females for oviposition at dosages of 0.5 g/100 ml water and above with the floating-cap method. The activity of this isomer increased 50-fold when it was applied directly to the water surface. The other three isomers, (+)-(5S,6R), (+)-(5R,6R), and (–)-(5s, 6s), were not active. The active isomer was ovipositionally attractive not only toC. quinquefasciatus but also toC. tarsalis Coquillett; however, it was 100 times more active in the former than in the latter species.Aedes aegypti (L.) andAnopheles quadrimaculatus Say were not attracted to the pheromone, thus suggesting that the pheromone is genus-specific.     

Identification of Epoxyhenicosadiene and Novel Diepoxy Derivatives as Sex Pheromone Components of the Clear-Winged Tussock Moth Perina nuda

Four EAG-active components (A–D) were found in the solvent extract of virgin females of the clear-winged tussock moth, Perina nuda. The most abundant component (B, ca. 250 ng/female) was identified as (3Z,6S,7R,9Z)-6,7-epoxyhenicosa-3,9-diene by GC-MS analyses of the extract, chemical derivatization, and comparative chiral HPLC. Minor components also elucidated were (3Z,9Z)-cis-6,7-epoxyicosa-3,9-diene, (A); (3R,4S,6S,7R,9Z)-3,4-6,7-diepoxyhenicos-9-ene, (C); and its 3S,4R,6S,7R isomer, (D); with amounts of 0.4, 5, and 8 ngt/female, respectively. Component B showed weak attractiveness to male moths in the field. The attractiveness was significantly enhanced by addition of component(s) C and/or D. No males were captured with either the antipode of component B or its mixtures with the minor components. In this field test, noctuid Hypocala rostrata males were also attracted with the synthetic P. nuda pheromone.     

Identification Of Sex Pheromone Components Of The Painted Apple Moth: A Tussock Moth With A Thermally Labile Pheromone Component

The sex pheromone of the painted apple moth, Teia anartoides (Lymantriidae) was investigated using GC-EAD and GC-MS analysis, derivatization, TLC analysis, and field cage and field trapping bioassays. The major sex pheromone components were identified as (6Z,9Z)-henicosa-6,9-dien-11-one and (6Z,9Z)-henicosa-6,9-diene. Other minor components of pheromone gland extracts included (6Z)-9R,10S-epoxyeicos-6-ene, (6Z)-9R,10S-epoxyhenicos-6-ene, (6Z,9Z)-henicosa-6,9-dien-11-ol, (6Z)-henicos-6-en-11-one, and (6Z, 8E)-henicosa-6,8-dien-11-one, but the roles of these minor components remain equivocal. In field cage and field experiments, a blend of all seven identified components [(6Z,9Z)-henicosa-6,9-dien-11-one (relative amount 100), (6Z,9Z)-henicosa-6,9-diene (100), (6Z)-9R,10S-epoxyeicos-6-ene (5), (6Z)-9R,10S-epoxyhenicos-6-ene (10), (6Z,9Z)-henicosa-6,9-dien-11-ol (5), (6Z)-henicos-6-en-11-one (1), and (6Z,8E)-henicosa-6,8-dien-11-one (25)] was as attractive to males as calling females, but tests with blends of the major component(s) with subsets of the minor components did not produce consistent results that unequivocally showed the various minor components to be critical components of the active blend. (6Z,9Z)-henicosa-6,9-dien-11-one is thermally labile and rearranges to (6Z,8E)-henicosa-6,8-dien-11-one and other products at ambient temperature, rendering the synthetic pheromone lure inactive after two days of field exposure.     

Laboratory and Field Study of the Attraction of Male Pea Midges, Contarinia pisi, to Synthetic Sex Pheromone Components

Behavioral activity of the recently identified sex pheromone components of the pea midge, Contarinia pisi, (2S,11S)-diacetoxytridecane, (2S,12S)-diacetoxytridecane, and 2-acetoxytridecane, was tested in wind tunnel and field-trapping experiments. In the wind tunnel, the attractancy of the three-component blend in a 7 : 10 : 0.1 ratio (following the above order, mimicking the ratios found in gland extract) did not differ significantly from female gland extract, whereas a mixture of the two major components (7 : 10) only attracted 2% of the males to the source. In the field, traps baited with the three-component blend caught by far the largest number of males. Traps baited with the two major components only caught slightly more than the blank traps, and catches in traps baited with 2-acetoxytridecane alone did not differ from catches in the blank traps. Traps baited with the racemate of all three components did not catch more than the blank traps, indicating that some of the enantiomers are inhibitory.     

Comparative Responses of Parasitoids to Synthetic and Plant-Extracted Nepetalactone Component of Aphid Sex Pheromones

The responses of aphid parasitoids to completely synthetic and plant-extracted nepetalactone, a component of aphid sex pheromones, were investigated. In wind tunnel tests, Praon volucre and Aphidius ervi responded equally strongly to both plant-extracted and 99% pure synthetic (4aS,7S,7aR)-nepetalactone. In the field, aphid-infested cereal trap plants were used to test the attractiveness of plant-extracted and synthetic (4aS,7S,7aR)-nepetalactone, as well as the synthetic enantiomer (4aR,7R,7aS)-nepetalactone. There was no significant difference in parasitisation levels by Praon spp. between plants baited with the synthetic or with the plant-extracted 7S isomers. However, the 7R isomer was unattractive alone and rendered the synthetic 7S isomer unattractive when combined to form a 50% blend of the two. It is concluded that the response of aphid parasitoids to nepetalactone is influenced more by enantiomeric purity rather than the possible presence of plant-related contaminants associated with plant extraction. The results are compared with data on male aphid responses to the compounds, which indicate that plant-derived contaminants can reduce attractancy, and the implications for the manipulation of parasitoids in the field are discussed.     

Optical isomers of 3,13-dimethylheptadecane: Sex pheromone components of the western false hemlock looper,Nepytia freemani (Lepidoptera: Geometridae)

(3S, 13R)-3, 13-Dimethylheptadecane [(3S, 13R)-3, 13-dime-17Hy] is the major pheromone component of the western false hemlock looper (WFHL),Nepytia freemani. In comparative gas chromotographic-electroantennographic detection (GC-EAD) analyses of stereoselectively synthesized isomers, 1 pg of (3S, 13R)-dime-17Hy elicited significantly stronger electrophysiological responses by male WFHL antennae than did 1 pg of separately injected (3R, 13R)-, (3R, 13S)- or (3S, 13S)-3, 13-dime-17Hy. In field experiments with individually tested stereoisomers. (3S, 13R)-3, 13-dime-17Hy was the only stereoisomer to attract males, but the four-stereoisomer blend was 3.6 times more attractive. Quaternary and all binary combinations of (3S, 13R)-3, 13-dime-17Hy with the other stereoisomers were equally attractive, suggesting that synergisytic behavioral activity in WFHL resided with either one of (3R, 13R)-, (3R, 13S)-, or (3S, 13S)-3, 13-dime-17Hy. Because optical isomers of (di)methylhydrocarbons do not separate on currently available columns, it remains unknown whether female WFHL also produce a four-stereoisomer pheromone blend. Substitutionality of pheromone stereoisomers without loss of behavioural activity has not previously been reported, but favorably compares with the concept of pheromone redundancy that was first suggested for the multiple pheromone component blend of the cabbage looper moth,Trichoplusia ni.     

Sex Pheromone Components of Indian Gypsy Moth, <Emphasis Type="Italic">Lymantria obfuscata</Emphasis>

The Indian gypsy moth, Lymantria obfuscata (Lepidoptera: Lymantriidae), has been recognized as a distinct species since 1865 but closely resembles a diminutive form of gypsy moth, Lymantria dispar. We tested the hypothesis that the sex pheromones of L. obfuscata and L. dispar are similar. In laboratory mate acceptance studies, very few male L. dispar made copulatory attempts when paired with female L. obfuscata, suggesting that female L. obfuscata emit one or more pheromone components antagonistic to male L. dispar. In coupled gas chromatographic–electroantennographic detection (GC–EAD) analyses of pheromone gland extract of female L. obfuscata, (Z)-2-methyloctadec-7-ene (2Me-7Z-18Hy) and (7R,8S)-cis-7,8-epoxy-2-methyloctadecane [(+)-disparlure] were most abundant and elicited the strongest responses from male L. obfuscata antennae. In field experiments near Solan (Himachal Pradesh, India), 2Me-7Z-18Hy and (+)-disparlure in combination attracted more male L. obfuscata than did either component alone. This two-component sex pheromone contrasts with the single-component sex pheromone [(+)-disparlure] of L. dispar. The contrasting composition of the lymantriid communities inhabited by L. obfuscata and L. dispar may explain why 2Me-7Z-18Hy is a pheromone component in L. obfuscata and a pheromone antagonist in L. dispar and why (−)-disparlure reduces pheromonal attraction of male L. dispar but not male L. obfuscata.     

Responses of Japanese Giant Looper Male Moth to Synthetic Sex Pheromone and Related Compounds

The Japanese giant looper, Ascotis selenaria cretacea, is a serious defoliator of tea gardens in Japan. In Mie Prefecture male moths were dose-dependently attracted to rubber septa baited with 0.1–10.0 mg of the sex pheromone component (Z,Z)-6,9-cis-3,4-epoxynonadecadiene. Monitoring by pheromone traps showed three flight periods (June, July, and August–September). A septum with a 1-mg dose was still active after exposure for over three months in the field, but the number of males captured by the old lure was one fifth of that captured by a fresh lure. The parent 3,6,9-triene, a minor pheromonal component, showed a synergistic effect on male attraction when mixed with the pheromonal epoxydiene at a very low ratio, but the 1:1 mixture hardly captured any male moths. Attraction was also effectively inhibited by the positional isomers (6,7- and 9,10-epoxydienes), which exhibited rather strong electrophysiological activities on the male antennae. When these related compounds were placed around a trap baited with the synthetic pheromone, the number of captured moths remarkably decreased, which indicates the possibility of using the compounds as a disruptant.     

Bioactivity, Synthesis, and Chirality of the Sex Pheromone of Currant Stem Girdler, Janus integer

It was previously reported that females of the currant stem girdler, Janus integer Norton (Hymenoptera: Cephidae), produce a compound, (Z)-9-octadecen-4-olide (1), that is sensitively detected by the antennae of males only. These characteristics suggested a pheromonal function, and this has now been confirmed with behavioral tests. Field tests conducted during two seasons in a commercial red currant field in Washington State showed that synthetic racemic 1 is attractive to male J. integer under natural conditions. A clear dose-response was evident, with greatest numbers of girdlers caught in sticky traps baited with 10 mg of the pheromone (in rubber septa) and least in traps baited with 1 mg or less. During May 2002, 10, 5, 3, and 1 mg baited traps caught means of 41.4, 26.6, 6.7, and 2.7 males/trap/visit (3–5 day intervals), respectively, with a maximum of 229 males caught in a single trap baited with 5 mg. A new synthetic method for racemic 1 is presented. The absolute configuration of natural 1 from the male sawflies was determined to be (R). The potential for using the sex pheromone of J. integer to improve management of this currant and gooseberry pest is discussed.     

Field evaluation of synthetic compounds mediating oviposition inCulex mosquitoes (Diptera: Culicidae)

3-Methylindole (skatole), a compound shown to be an oviposition attractant/stimulant forCulex quinquefasciatus in laboratory tests, was evaluated against natural populations of mosquitoes in the field. In experiments using paired black plastic tubs that contained water treated with an attractant solution containing a mixture of 3-methylindole, 4-methylphenol, 4-ethylphenol, phenol, and indole or that contained only tap water, the attractantbaited traps received significantly moreCx. quinquefasciatus egg rafts than did the untreated tap water. Gravid female traps that were baited with attractant solution collected significantly moreCx. quinquefasciatus females than did traps containing only tap water. There was no significant difference in number ofCx. quinquefasciatus collected in the traps baited with the attractant solution compared to the traps that contained water with 3-methylindole alone, indicating that 3-methylindole was solely responsible for the attraction. In replicated experiments conducted in experimental ponds, ponds that were treated with 3-methylindole received significantly moreCx. quinquefasciatus, Cx. tarsalis, andCx. stigmatosoma egg rafts than did untreated ponds. Experimental ponds treated with 3-methylindole at two levels (0.12 and 0.6 mg/liter) were equally attractive to ovipositingCx. quinquefasciatus. This is the first record of 3-methylindole showing attractancy/stimulation to ovipositingCx. stigmatosoma andCx. tarsalis under field or laboratory conditions.     

Sex Pheromone Components of Casuarina Moth, Lymantria xylina

cis-7,8-Epoxy-2-methyleicosane is a sex pheromone component of the Casuarina moth, Lymantria xylina Swinhoe. The compound was extracted from pheromone glands of female moths and was identified by coupled gas chromatographic–electroantennographic detection (GC-EAD) and GC–mass spectrometry. In field experiments in Taiwan, traps baited with either or both of (7R,8S)-cis-7,8-epoxy-2-methyleicosane (>99% ee) [termed here (+)-xylinalure] and (7S,8R)-cis-7,8-epoxy-2-methyleicosane (>99% ee) [termed here (–)-xylinalure] captured male L. xylina. Addition of further candidate pheromone components to xylinalure did not enhance its attractiveness. Demonstration of whether or not female L. xylina produce both optical isomers of xylinalure, and determination of the ratio, will require pheromone extract analyses on a chiral, enantiomer-separating column (as yet unavailable) or derivatization of epoxides in accumulated gland extracts. Attraction of male L. xylina to either enantiomer of xylinalure contrasts with enantiospecific production of, and/or response to, epoxy pheromones in congeners. With no other nocturnal lymantriid moth known in Taiwan to utilize xylinalure for pheromonal communication, enantiospecific fine tuning of xylinalure, or evolution of a more complex pheromone blend, may not have been necessary for L. xylina to maintain specificity of sexual communication. Racemic xylinalure will be appropriate for pheromone-based detection surveys of L. xylina in North America.     

Identification of the Sex Pheromone of the Swede Midge, <Emphasis Type="Italic">Contarinia nasturtii</Emphasis>

Coupled gas chromatographic–electroantennographic detection analyses of ovipositor extracts of calling Contarinia nasturtii females revealed two compounds that elicited responses from antennae of male midges. Using synthetic reference samples, these components were identified by gas chromatography–mass spectrometry and enantioselective GC as (2S,9S)-diacetoxyundecane and (2S,10S)-diacetoxyundecane. In addition, trace amounts of 2-acetoxyundecane were found in ovipositor extracts, and the (S)-enantiomer was synthesized. When tested in the wind tunnel, a blend of 5ng (2S,9S)-diacetoxyundecane and 10 ng (2S,10S)-diacetoxyundecane (mimicking the ratio found in the extracts) did not attract any of the males tested, but when 0.1 ng (S)-2-acetoxyundecane was added to the blend, 86.8% of the males were attracted to the bait. Three-component blends with lower or higher relative concentrations than 1% of (S)-2-acetoxyundecane [relative to (2S,10S)-diacetoxyundecane] were less attractive. In a field trapping experiment with released laboratory-reared C. nasturtii adults, traps baited with 500:1000:10 ng of (2S,9S)-diacetoxyundecane/(2S,10S)-diacetoxyundecane/(S)-2-acetoxyundecane applied to rubber septa or dental cotton rolls were tested. Traps without dispensers were used as controls. All three treatments were tested at 20 and 50 cm above ground. The estimated recapture rate was 30–50%, and 81.9% of the recaptured males were caught in traps positioned at 20 cm above ground, and 88.4% in traps with dental cotton rolls as dispensers.     

2-Methyl-(Z)-7-Octadecene: Sex Pheromone of Allopatric Lymantria lucescens and L. serva

Our objective was to identify the sex pheromone of Lymantria lucescens and Lymantria serva (Lepidoptera: Lymantriidae), whose larvae defoliate, respectively, Quercus spp. in temperate regions and Ficus spp. in the subtropics. Coupled gas chromatographic-electroantennographic (GC-EAD) detection analyses of pheromone gland extracts revealed one EAD active compound produced by female L. lucescens and by female L. serva. This was identified as 2-methyl-(Z)-7-octadecene (2me-Z7-18Hy) by retention index calculations on DB-5, DB-23, and DB-210 columns and by comparative GC-mass spectrometric (MS) and GC-EAD analyses of the insect-produced candidate pheromone and synthetic 2me-Z7-18Hy. In field experiments, traps baited with 2me-Z7-18Hy captured male L. lucescens near Toyota City, Japan, and male L. serva in Taipei, Taiwan. Allopatric distribution of L. lucescens and L. serva seems to allow both species to use the same sex pheromone without compromising its specificity.     

Pheromone Components and Diel Periodicity of Pheromonal Communication in Lymantria fumida

Extracts of pheromone glands from female Lymantria fumida were analyzed by coupled gas chromatographic–electroantennographic detection (GC-EAD) and by coupled GC–mass spectrometry (MS). The two compounds that elicited responses from male L. fumida antennae were identified as cis-7,8-epoxy-2-methyloctadecane (disparlure) and 2-methyl-Z7-octadecene (2me-Z7–18Hy). Field experiments in northern Japan demonstrated that synthetic (7R,8S)-cis-7,8-epoxy-2-methyloctadecane [(+)-disparlure] and 2me-Z7–18Hy are synergistic sex pheromone components of L. fumida. (7S,8R)-cis-7,8-Epoxy-2-methyloctadecane [(–)-disparlure] had no behavioral effect on male L. fumida. Traps baited with (+)-disparlure and 2me-Z7–18Hy captured male L. fumida between 21:00 and 24:00 hr, whereas traps baited with (+)-monachalure [(7R,8S)-cis-7,8-epoxy-octadecane], (+)-disparlure and 2me-Z7–18Hy attracted males of the nun moth, L. monacha L., between 02:00 and 04:00 hr. Both temporal separation of pheromonal communication and specificity of pheromone blends seem to contribute to the reproductive isolation of sympatric and coseasonal L. fumida and L. monacha.