A comparative study of the phytotoxic effects of siricid woodwasps on conifers

The effects of siricid mucus and the symbiotic fungus injected artificially, or by oviposition of caged females on young conifer trees were investigated in a comparative study of the phytotoxicity caused by seven species of siricid woodwasps. Dilute solutions of mucus were bioassayed using detached shoots. Rapid physiological changes in the radial growth of the stem, quantity of starch in the leaves, leaf pressure and colour of the foliage of living trees were induced by only one species, Sirex noctilio. Solutions of S. noctilio and S. cyaneus mucus caused premature senescence of needles on detached shoots. The other species, Sirex juvencus, Urocerus gigas, U. augur, U. sah and Xeris spectrum , produced no phytotoxic symptoms in living trees and less rapid senescence of needles on detached shoots. These differences are discussed in relation to the ecology of siricids in the northern hemisphere and the successful establishment of S. noctilio in pine plantations of Australasia.     

Experimental studies on the responses of European siricid woodwasps to host trees

A study was made of the oviposition responses of the siricid woodwasps, Sirex noctilio, S. juvencus, S. cyaneus and Urocerus gigas to experimentally debilitated spruce and pine trees. Treatments included felled, fire-burned, logged and girdled trees which were treated at intervals before bioassay with cages siricid females. In comparable treatments, S. noctilio showed a distinct preference for pines although it was quite successful on spruce while the other siricid species displayed a preference for spruce. The Sirex species successfully attacked more recently treated material although U. gigas showed a preference for timber in a more advanced state of debility. All species showed preferences for girdled trees although other treatments were also favoured by the different siricid species. Overall, the siricids preferred the area above the girdle irrespective of tree species and there was a higher rate of eggs above the girdle in treated spruce trees. A wide range of moisture content in the timber appeared to have no effect on drilling activity or the successful development of eggs. The response of a wild population of S. juvencus to the treated spruce is also described. The results are discussed in relation to the ecology of Siricidae in Europe and the establishment of S. noctilio in Australia. It is suggested that of the species studied, only S. noctilio poses a serious threat to the living pine stands of Australia.     

DNA sequence and RFLP data reflect geographical spread and relationships of Amylostereum areolatum and its insect vectors

The white rot fungus, Amylostereum areolatum (Basidiomycetes), is best known for its symbiotic relationship with various siricid wood wasp species. In this study, the relationship between isolates of A. areolatum associated with two wood wasp species, Sirex noctilio and S. juvencus, are considered to identify possible intraspecific groups. Isolates from the northern (native) and southern (exotic) hemispheres are included to determine patterns of geographical spread and origin of introductions into the southern hemisphere. The phylogenetic relationships of these isolates to authentic isolates of A. chailletii, A. laevigatum and A. ferreum were also investigated. Sequence and restriction fragment length polymorphism (RFLP) analyses of the variable nuc-IGS-rDNA region provided markers to distinguish intraspecific groups within A. areolatum. Isolates of A. areolatum associated with S. noctilio and S. juvencus contained four heterogenic sequences in the DNA region analysed. These sequences occurred in one of five combinations in each isolate. Some of these sequences were unique to isolates of A. areolatum from either wasp species, while others were present in both groups. This shows the ancient and specialized evolutionary relationship that exists between these insects and fungi. Isolates from the southern hemisphere all share the same sequence group. This supports previous hypotheses that S. noctilio has spread between countries and continents of this region. At the interspecific level, the IGS-rDNA sequence analysis showed that A. ferreum and A. laevigatum are closely related to each other, and they in turn are related to A. chailletii. Amylostereum areolatum was the most distinctly defined species in the genus. This can be attributed to the obligate relationship between A. areolatum and its insect vectors. Polymerase chain reaction-RFLP analysis was also shown to be an effective tool to distinguish between the different species of Amylostereum.     

Reproductive strategy of a woodwasp with no fungal symbionts, Xeris spectrum (Hymenoptera: Siricidae)

Experiments were conducted to elucidate the reproductive strategy of the siricid woodwasp, Xeris spectrum, which carries no substantial symbiotic fungi in its body, in a comparison with the life cycles of two fungus-carrying siricid woodwasp species, Sirex nitobei and Urocerus japonicus, by considering ecological traits such as seasonal patterns of occurrence, spatial distribution of emergence on a tree, and oviposition activities. Part of the X. spectrum populations emerged in spring, during May and June, while others emerged in summer, during August and September, simultaneously with other siricid fungus-carrying woodwasps. The vertical distribution pattern of X. spectrum emergence holes on the trunk closely coincided with the emergence hole pattern of S. nitobei. X. spectrum laid few eggs on fresh logs, old logs, or on logs inoculated with potato dextrose agar, whereas on logs inoculated with Amylostereum chailletii or A. areolatum, X. spectrum females oviposited no less than 30%, on average, of their potential eggs. Moreover, the oviposition sites on these logs were concentrated near the Amylostereum inoculation positions. These results indicate that X. spectrum has evolved a life history that utilizes fungal symbionts of other woodwasp species without itself possessing any symbiotic fungus. Moreover, X. spectrum has evolved a dual reproductive system in that (1) some adults emerge in summer during the same emergence period as the fungus-carrying woodwasps and thereby oviposit on host trees already inoculated with fungi, and (2) other adults emerge the next spring and oviposit on trees that were inoculated with fungi a year earlier. Received: 16 January 1997 / Accepted: 14 July 1997     

Within-tree distributions of the Sirex noctilio Fabricius (Hymenoptera: Siricidae)--parasitoid complex and development of an optimal sampling scheme

Little is known about the introduced European woodwasp, Sirex noctilio F. (Hymenoptera: Siricidae), and its hymenopteran parasitoid complex in North America. To assess within-tree and landscape-level densities and distributions of these insects, and develop a more cost effective approach for sampling and monitoring S. noctilio, 18 infested pines (12 P. resinosa Aiton and six P. sylvestris L.) were felled from six stands within three regions in central New York and cut into 0.5-m bolts. Bolts were carefully split to recover all siricids (S. noctilio and native siricids) and parasitoids. In total, 2,558 siricids were recovered; 1,972 siricid larvae and 586 teneral adult S. noctilio. Parasitism of siricids, a majority of which were S. noctilio based on larval rearing results, was 16.4% with Ibalia leucospoides ensiger Norton causing 10.7% of the documented mortality. Numbers of siricids and parasitoids declined 33-86% from the northern to the southern sampling regions, peak insect densities occurred in sections of the bole 15-19 cm in diameter and numbers of insects were generally higher in P. sylvestris than P. resinosa according to the highest ranked zero-inflated poisson and probit regression models. Bark thickness was not correlated with siricid or parasitoid species densities. We also describe sampling plans where as few as two 0.5-m samples from infested trees provided reliable within-tree insect densities.     

松树蜂与其共生真菌的互利共生关系

松树蜂Sirex noctilio Fabricius是一种重要的国际林业检疫性害虫,主要危害针叶树,原产欧亚大陆和北非。近100多年来,先后入侵大洋洲(新西兰和澳大利亚)、南美洲(乌拉圭、阿根廷、巴西和智利)、北美洲(加拿大和美国),以及南非。2013年8月,在中国黑龙江省内首次发现松树蜂,目前发现其主要危害樟子松。松树蜂能与一种淀粉韧革菌属Amylostereum的真菌Amylostereum areolatum(Fr.)Boidin形成严格的互利共生关系,该虫除直接钻蛀树木外,还能通过产卵行为将自身毒素腺体分泌的毒素和体内共生真菌随同虫卵一起注入寄主树木体内,形成"虫-毒-菌"3个致害因子相互协作的特殊危害方式,加速树势的衰弱并造成寄主树木死亡。本文就国内外松树蜂与其共生菌互利共生关系的研究进行了综述,分别从结构与功能的层次上对其互利共生关系进行了梳理和总结,重点阐释了松树蜂与共生菌的营养共生关系,松树蜂携带传播共生菌的机制,共生菌的种群遗传学以及松树蜂毒素和共生菌在危害寄主树木时的协同关系等。以期为开展关于松树蜂的专项研究提供一些合理的建议,同时为积极有效地防控该害虫提供科学依据。     

The responses of Ibalia species (Hymenoptera: Ibaliidae) to the fungal symbionts of siricid woodwasp hosts

The oviposition response of Ibalia drewseni and Ibalia leucospoides is described and compared. Oviposition behaviour is elicited by odours produced by the symbiotic fungus of several species of siricid woodwasps. The responses of the two Ibalia species to fungus cultures of different ages and host sources is discussed in relation to their emergence periods and the ecology of their hosts.     

Host-marking by female pepper weevils, Anthonomus eugenii

Pepper weevils, Anthonomus eugenii Cano (Coleoptera: Curculionidae), feed and oviposit in flower buds and small fruits of plants in the genus Capsicum, as well as several species of Solanum (Solanaceae). Females chew a small hole into the fruit, deposit a single egg within the cavity, and seal the hole with a clear anal secretion that hardens into an ‘oviposition plug’. Female oviposition behavior was studied in a series of small‐arena bioassays to determine whether previous oviposition in Jalapeño pepper fruit deterred subsequent oviposition and to determine what specific cues from an infested fruit influence female behavior. In choice and no‐choice tests, females preferred clean fruit to fruit that had received four eggs 24 h previously (i.e., infested fruit), whether the fruit was infested with conspecific eggs or their own eggs. Further bioassays demonstrated that the presence of female frass, or oviposition plugs alone, in the absence of eggs or any fruit damage, was sufficient to deter oviposition. In addition, females given the choice between an infested fruit with the oviposition plug removed or an unaltered infested fruit preferred the fruit with no plugs, even when eggs, frass, and feeding damage were still present. To determine whether females would avoid infested peppers under more natural conditions, we quantified oviposition on infested and uninfested sentinel pepper fruit within individually caged plants and on clean and infested plants caged together. Females consistently laid more eggs on clean fruit than on infested fruits and moved within and among pepper plants to search for more acceptable oviposition sites. We conclude that oviposition plugs, along with contaminated female, but not male, frass contain a deterrent that, in the absence of any other cue, is enough to alert a female that a patch is occupied.     

Ovipositional behaviour of Simulium argyreatum and S. variegatum and its relationship to infection by the fungus Erynia conica (Entomophthoraceae)

SUMMARY. 1. At two sites in Devon S. argyreatum and S. variegatum only oviposited on water-splashed boulders. The oviposition season was from 15 March to 9 November at Steps Bridge and from 28 March to 7 December at Fingle Bridge. Flies laid eggs only during a brief period after sunset.
2. Flies infected with the fungus E. conica returned to the oviposition sites at the same times as healthy flies, but infected flies appeared to be unable to lay eggs.
3. At least 10.9% of healthy flies were found to have infective conidia of E. conica on their wings after laying eggs.     

The fecundity, development and host relationships of Ceratobaeus spp. (Hymenoptera: Scelionidae), parasites of spider eggs

Abstract. 1. Ceratobaeus spp. enter the nest of their host spider and oviposit into eggs through the thin silk eggsac.
2. Temperatures below 15° C limit oviposition by slowing parasites down.
3. Males emerge prior to females and mate with their sibs. Sex ratios of 6.6–6.0:1 in favour of females were observed in the field and laboratory for two species.
4. Even though females can oviposit almost immediately after emergence their full complement of eggs (= 65) is not reached until several days later.
5. Superparasitism is low and appears to occur from subsequent accidental ovipositions. No marking of the external surface of hosts was observed.
6. Parasites overwinter as adults under bark. They do not feed as adults nor do they reabsorb their eggs, but rather oviposit as soon as host eggs become available in spring.
7. Successful oviposition occurs in later stages of host eggs reared at 15° C and 20° C than it does at 25°C. Different rates of development between host and parasite is proposed as an explanation for this phenomenon.
8. The species studied show varying degrees of specificity but each has a dominant host. Location of hosts involves cues from the habitat (bark), silk nests of spiders, and some factor associated with host eggs.     

Repeated copulation in the brown planthopper, Nilaparvata lugens Stál (Homoptera; Delphacidae)

Abstract. 1. Mating of the brown planthopper, Nilaparvata lugens , was investigated in relation to oviposition. Females became unreceptive immediately after mating and showed various types of repelling behaviour to courting males. As a result, females usually did not mate repeatedly in quick succession, but after ceasing to lay fertilized eggs they behaved as virgins and mated again before producing more fertilized eggs.
2. Copulation, followed by deposition of fertilized eggs, occurred twice, or in a few cases three times, throughout the adult stage. Copulation lasted about 2 min at the first mating and about 1 min at the second or third mating.
3. When the number of fertilized eggs began to decrease rapidly, oviposition rate also decreased, but it increased again immediately after re-mating. Repeated copulation was also related to the potential rate of population increase. In the present experiments, the estimates of net reproductive rate ( R ₀) in brachypterous females was 287.7 (first mating), 88.3 (second mating) and 7.2 (third mating) and in macropterous females 286.8, 120.6 and 3.6 respectively.
4. In a patchy environment, repeated copulation following an invasion may contribute to the increase of a population.     

Oviposition on host plants by a tropical forest grasshopper (Microtylopteryx hebardi: Acrididae)

Abstract. 1. The tropical forest grasshopper Microtylopteryx hebardi Rehn (Orthoptera, Acrididae, Ommatolampinae) oviposits inside leaves or stems of its host plants. This is the first known instance of oviposition inside living plant tissue (rather than in the ground, on top of leaves, or in detritus) for a tropical forest acridid.
2. In understorey secondary forest at La Selva, Costa Rica, M.hebardi females fed on fifteen species of host plants, but oviposited in only four of these.
3. In laboratory choice tests, females showed selectivity for oviposition host plants similar to that inferred from field observations.
4. Plant volume was a better predictor of the number of eggs laid per stem than variables that indicated leaf or stem characteristics of individual plants.
5. Unlike the ovipositor of acridids laying eggs in the ground, that of M.hebardi is equipped with toothed, sclerotized edges, probably used to bore into plant tissue.
6. Endophytic oviposition in M.hebardi may be related to finding host plants in a habitat that is complex, heterogeneous, and rich in plant species.     

Effect of Two Bark Beetle-Vectored Fungi on the On-Host Search and Oviposition Behavior of the Introduced Woodwasp Sirex noctilio (Hymenoptera: Siricidae) on Pinus sylvestris Trees and Logs

Sirex noctilio’s fungal symbiont, Amylostereum areolatum, is required for its offspring’s development. The symbiont is a weak competitor with bark beetle-vectored fungi so it would be beneficial to the woodwasp if it avoided ovipositing in substrate colonized by these competitors. The response of S. noctilio to the presence of two beetle-vectored fungi, Leptographium wingfieldii and Ophiostoma minus, inoculated into living trees, and to L. wingfieldii and A. areolatum inoculated into cut logs was investigated. The wasp avoided areas with L. wingfieldii; there were fewer signs of oviposition activity and drilling in these zones. There was no significant response to O. minus or A. areolatum. Female woodwasps can detect the presence of some fungi and make choices about oviposition sites that benefit their offspring.     

Natural enemies associated with Sirex noctilio (Hymenoptera: Siricidae) and S. nigricornis in Ontario, Canada

Sirex noctilio F. is an exotic woodwasp now found in eastern North America where it shares natural enemies with native woodwasps of Pinus spp. To study the extent to which native hymenopteran parasitoids and parasitic nematodes could affect woodwasp populations, 60 Pinus trees with symptoms of S. noctilio attack were felled in 2007 and 2008 in Ontario, Canada. Each tree bole was cut into 1-m sections that were placed in individual rearing tubes; emergence was monitored from May to November of the year of felling. Female S. noctilio were dissected to assess parasitism by the nematode Deladenus siricidicola Bedding. Two species of Siricidae emerged from these trees; S. noctilio, which accounted for most of the specimens collected, and S. nigricornis F. Of the three species of parasitoid that emerged, Ibalia leucospoides (Hochenwarth) was the most abundant, accounting for an overall hypothetical Siricidae parasitism rate of almost 20%. This parasitoid emerged over a similar time period as S. noctilio-between early July and early September. Except in trees >15 m in height, parasitism by I. leucospoides generally appeared uniform throughout the bole. Parasitism rates did not vary between the 2 yr, but did between sites in 1 yr. Parasitic nematodes were found in the haemocoel of about one third of S. noctilio females dissected but were never found sterilizing the eggs; none were found in S. noctilio emerging from P. resinosa. These findings suggest that I. leucospoides is currently the primary invertebrate natural enemy of S. noctilio in Ontario.     

Oviposition preference in the oligophagous tomato fruit fly, Neoceratitis cyanescens

We assessed the role of visual and olfactory cues on oviposition preference in the oligophagous tomato fruit fly, Neoceratitis cyanescens (Bezzi) (Diptera: Tephritidae). In a field survey, we evaluated the stage of susceptibility of field‐grown tomatoes by monitoring N. cyanescens infestations from fruit‐setting up to harvest, in relation to post‐flowering time, size, and visual properties of fruit. In two‐choice laboratory experiments, we tested the degree to which females use visual and olfactory cues to select their host plant for oviposition. In addition, we investigated the ability of flies to avoid fruit already infested by conspecific eggs or larvae, and the influence of natal host fruit on oviposition preference. Neoceratitis cyanescens females preferentially lay their eggs in small yellow‐green unripe fruit (2–3.5 cm diameter, 10–21 days post‐flowering). Damage to fruit was significantly affected by brightness and size properties. In laboratory experiments, females chose to lay their eggs in bright orange rather than yellow domes. On the sole basis of olfactory stimuli, females showed a significant preference for unripe vs. ripe host fruit, for unripe fruit vs. flowers or leaves, and for host vs. non‐host fruit (or control). However, colour interacted with odour as females dispatched their eggs equally between the yellow dome and the bright orange dome when unripe fruit of tomato was placed under the yellow dome vs. ripe fruit under the bright orange dome. When offered real ripe and unripe tomatoes, females preferred unripe tomatoes. Females significantly chose to lay eggs in non‐infested fruit when they were given the choice between these or fruit infested with larvae. In contrast, recent stings containing eggs did not deter females from laying eggs. Rather, they could have an attractive effect when deposited within <1 h. Regardless of their natal host plant, tomato or bugweed, N. cyanescens females laid significantly more eggs in a dome containing bugweed fruit. However, 15% of females originating from tomato laid eggs exclusively in the dome with tomato, against 3% of females originating from bugweed.     

Influence of egg size differences within egg clutches on larval parameters in nine libellulid species (Odonata)

Abstract.  1. In libellulids, egg size differs between species and populations. There are also size differences within egg clutches of individual females.
2. Past experiments suggest that there are two different types of egg clutches in libellulids. Egg size decreases significantly during oviposition in species that perform non-contact guarding during oviposition. In contrast, in species ovipositing in tandem, egg size is randomly distributed.
3. This study deals with the possible consequences of egg size variation within the different egg clutch types. The study examined whether there is a correlation between egg development time, offspring sex or larval size and egg size.
4. The current experiments were conducted in Namibia and Germany. Five non-contact guarding and four tandem guarding libellulid species were used.
5. In some species larger eggs needed more time to develop, in some species no correlation between egg size and egg development time could be found, whereas in other species larger eggs developed faster.
6. The sex ratio was biased towards females in Leucorrhinia dubia and in Sympetrum striolatum and egg size was not associated with gender.
7. In both egg clutch types larger eggs resulted in larger larvae. In this study, evidence was found that the effects of egg size diminished with progressing larval development under good conditions. However, it is possible that the effects may have a greater influence under harsh circumstances.     

Dispersal by bagworm larvae, Metisa plana: effects of population density, larval sex, and host plant attributes

Abstract 1. The work reported here investigated the incidence of dispersal by bagworm larvae Metisa plana (Walker) (Lepidoptera: Psychidae), a species with apterous females and winged males.
2. A mark–recapture study conducted in a plantation of oil palm revealed that a significant proportion of larvae undertakes multiple episodes of ballooning, and suggested that ballooning represents a strategy for foraging both within and between hosts.
3. Experiments conducted in a controlled cage environment indicated that increasing population density and defoliation of oil palm both promote dispersal by larvae.
4. Neonatal larvae typically remained on the host where they emerged, indicating that pupation site of late-instar females influences the distribution of their progeny, and suggesting a high level of genetic relatedness in localised bagworm populations.
5. Density-dependent dispersal may generate relatively uniform distributions of M. plana in plantations of oil palm, by simultaneously stabilising populations on heavily infested palms and redistributing larvae on lightly infested palms.
6. The rate of ballooning was greater for female than for male larvae, possibly resulting from the sex-specific impact of population density on reproductive success or from late-instar females seeking suitable oviposition sites for their future progeny.     

Gallery construction and oviposition of the bark beetle Ips typographus (Coleoptera: Scolytidae) at different breeding densities

Abstract. 1. The bole of a Norway spruce, Picea abies (L.) Karst., was cut in 25 cm sections and infested in the laboratory with 2 or 8 females/dm² of Ips typographus (L.) and an equal number of males.
2. One log of each beetle density was dissected every second day, beginning 2 days after female introduction.
3. The oviposition rate was highest at the beginning of the breeding period for both densities and then declined, especially at the high density. Residence time is a useful approximation for the number of laid eggs in the beginning of the oviposition period or when the density is low.
4. Final gallery length and egg number per gallery were on average significantly smaller at density 8 than at density 2.
5. Linear relationships between total gallery length and number of egg niches were found to be similar for the two densities.
6. An estimate of the egg to adult mortality at different densities was found by using data from a field study that related gallery length to breeding density in attacked living trees, together with published relationships between density and egg niches or emergence of new adults.
7. It was concluded that both decreased oviposition rate and earlier re-emergence at higher densities contribute to a declining oviposition with increasing density. The final offspring production is futher reduced by lower survival due to larval competition as density increases.     

松树蜂的产卵行为

【目的】研究松树蜂Sirex noctilio的产卵行为,明确其产卵能力,为评估其繁殖潜力和危害能力提供基础数据。【方法】于黑龙江省大庆市杜尔伯特蒙古族自治县新店林场采集被松树蜂危害的寄主樟子松Pinus sylvestris var.mongolica木段上获得虫源。在室内条件下观察和分析松树蜂产卵的行为过程及规律。解剖松树蜂在樟子松木段上的产卵孔,并观察其结构特征。【结果】松树蜂羽化时即性成熟,能够马上进行交配和产卵。松树蜂雌成虫一次完整的产卵过程主要分为4个动作:树皮钻孔、木质部钻刺、产卵(注入有毒黏液和共生真菌Amylosereum areolatum)和产卵器拔出。产卵时间在360～540 s之间的产卵频数最多,占产卵总频数的41.40%;产卵过程中木质部钻刺用时最长,至少占整个产卵过程用时的90%。松树蜂雌成虫在一个产卵孔处会进行1～4次产卵,产卵时间和产卵次数呈显著正相关,当产卵时间t<360 s时,进行了1次产卵;当360 s≤t<540 s时,进行了2次产卵;当540 s≤t<780 s时,进行了3次产卵;当t≥780 s时,进行了4次或更多次产卵。松树蜂在一个产卵孔处进行1, 2, 3和4次产卵的比例分别为21.66%, 41.40%, 27.39%和9.55%。【结论】松树蜂雌成虫在一个产卵孔处的产卵时间和产卵次数呈正相关。利用产卵时间和产卵次数的关系,在只调查产卵时间的情况下,可以推断产卵次数。松树蜂在一个产卵孔处产卵的次数多,对寄主樟子松危害大。     

Oviposition preferences of Maculinea alcon as influenced by aphid (Aphis gentianae) and fungal (Puccinia gentianae) infestation of larval host plants

Abstract 1. The influence of infestation of the larval host plant Gentiana cruciata on the egg‐laying preferences of the xerophilous ecotype of Alcon Blue butterfly (Maculinea alcon) was studied in a semi‐dry grassland area (Aggtelek Karst Region, Northern Hungary). 2. We examined whether oviposition patterns of females differed when G. cruciata stems were uninfested compared with when they were infested by an aphid (Aphis gentianae) or a rust (Puccinia gentianae) species. 3. Females laid more than 90% of their eggs on fertile, uninfested G. cruciata stems, although these stems comprised only ～ 50% of the total stems available. Stems infested by aphids were similar to uninfested ones in properties that had a strong correlation with egg numbers, and yet there were significantly fewer eggs on infested stems than on intact ones. 4. Females never laid eggs on parts of Gentiana stems infested by aphids, and the presence of Lasius paralienus ants, which have a mutualistic interaction with Aphis gentianae, did not increase the repulsive effect of aphids. Infection of Gentiana by Puccinia did not influence the egg‐laying behaviour of females, even though the flowers and buds of infested stems exhibited a delayed development. 5. Aphid infestation can influence butterfly oviposition patterns through both direct and indirect effects. The presence of aphids directly excluded oviposition, but our data also indicated the possibility of an indirect effect of aphid infestation. Stems that had no aphids at the last egg counting, but were infested prior to it, had significantly fewer eggs than those that were never infested.