A review of the natural enemies of beetles in the subtribe Diabroticina (Coleoptera: Chrysomelidae): implications for sustainable pest management

Diabroticina is a speciose subtribe of New World Chrysomelidae (Subfamily Galerucinae: Tribe Luperini) that includes pests such as corn rootworms, cucumber beetles and bean leaf beetles (e.g. Diabrotica, Acalymma, Cerotoma species). The evolution and spread of pesticide resistance, the European invasion of Diabrotica v. virgifera LeConte, and possible development of resistance due to the large-scale deployment of Diabrotica-active Bt maize in North America have generated a sense of urgency in developing biological control options against Diabroticina pests. In the present study, we review available knowledge on biological control options, including 290 publications on natural enemy–Diabroticina associations in the New World. Several natural enemy species or groups appear to be promising candidates for control strategies with different ecological rationales. We propose that future research should pursue: (1) development of inundative biological control products, particularly mass-produced entomopathogenic nematodes and fungi, (2) understanding of specific natural enemies of Diabroticina larvae throughout the Americas and of adults particularly in higher altitudes of Central America or northern South America including potential classical biological control agents against D. v. virgifera; (3) enhancement of natural enemies through cultural practices, i.e., reduced tillage, reduced weed control, cover crops, diversified crop rotations or soil amendments. Research and action must be coordinated to accelerate the exploration of biological control options.     

A critical evaluation of host ranges of parasitoids of the subtribe Diabroticina (Coleoptera: Chrysomelidae: Galerucinae: Luperini) using field and laboratory host records

The subtribe Diabroticina is a large group of New World Chrysomelidae that includes corn rootworms, cucumber beetles and other pests. Recent introductions of Diabrotica virgifera virgifera LeConte into Europe, and the development of resistances to current management practices of rootworms in the USA have increased interest in new sustainable options for managing those pests. The only parasitoids that have been shown to consistently target and develop inside the beetle adults are Centistes gasseni Shaw, Centistes diabroticae Gahan (both Hym.: Braconidae), and Celatoria diabroticae Shimer, Celatoria compressa (Wulp), Celatoria bosqi Blanchard, and Celatoria setosa Coquillett (all Diptera: Tachinidae). This review improves our understanding of the realised and potential host range of these known parasitoids by rectifying erroneous references in light of new host records and from laboratory host range tests. Based on this critical review, all tachinid and braconid species studied are considered to be specific at least to the level of subtribe, i.e. Diabroticina. Celatoria setosa, Celatoria diabroticae and C. bosqi, have a narrow realized and potential host range; the former is restricted to the genus Acalymma and the last two to the fucata and virgifera groups of the genus Diabrotica. The braconids Centistes gasseni and C. diabroticae are also specific. The realized host range of C. gasseni includes species in the Diabrotica fucata and virgifera groups; while its potential host range also includes Acalymma species. The realized and potential host range of Centistes diabroticae includes Acalymma species as well as species in the fucata and virgifera groups of Diabrotica. Celatoria compressa has the broadest realised range compared to the other species studied, since it was obtained from species in several genera of Diabroticina; and its potential host range may also include Old World Aulacophora species.     

Biological control of <Emphasis Type="Italic">Lantana camara</Emphasis> in South Africa: targeting a different niche with a root-feeding agent, <Emphasis Type="Italic">Longitarsus</Emphasis> sp.

The root-feeding flea beetle, Longitarsus sp. (Coleoptera: Chrysomelidae: Alticinae), was studied as a potential biological control agent for Lantana camara L. (Verbenaceae) in South Africa. Host range tests were carried out on 52 plant species in 11 families. Although 11 plant species, all in the family Verbenaceae, supported complete development of Longitarsus sp. during no-choice tests, the beetles showed very strong preferences for L. camara during paired-choice and multi-choice tests. The results confirm that the beetles have a narrow host range, and that under natural conditions they are highly unlikely to utilise plants other than L. camara. In the unlikely event that some of the Lippia spp. are attacked in the field, they are not expected to sustain populations of the flea beetle over time. Attributes that should enhance the biocontrol potential of Longitarsus sp. include: the adults are long-lived and highly mobile; and, the larvae cause extensive direct damage to the roots of L. camara, which could in turn expose the plants to soil-born pathogens. All indications are that Longitarsus sp. could make a substantial contribution to the biological control of L. camara in many countries around the world because the beetles pose no threat to non-target plant species and they damage a part of the plant (i.e. roots) not yet affected by any other agent species.     

Host suitability and preference studies of Trichogramma cordubensis (Hymenoptera: Trichogrammatidae)

Studies of host suitability and preferences of Trichogramma cordubensis Vargas and Cabello (Hymenoptera: Trichogrammatidae) were performed with eggs of six Lepidoptera (Noctuidae) species: Thysanoplusia orichalcea Fabricius, Peridroma saucia (Hübner), Xestia c-nigrum L., Phlogophora meticulosa (L.), Noctua pronuba (L.), and N. atlantica (Warren). Host suitability was studied by analysing separately the effects of the attacked host species and the influence of the rearing host species on different biological parameters of T. cordubensis. Host preference was analysed by offering eggs of two host species simultaneously to a single female wasp without previous oviposition experience (dual-choice tests). Results show that P. saucia, followed by P. meticulosa were the least suitable hosts for T. cordubensis, since on these species the preimaginal development of the parasitoids was significantly longer and, the number of parasitized eggs as well the number of offspring per female were significantly lower. Contrarily, T. cordubensis parasitized at a higher rate the eggs of the endemic non-target species, N. atlantica. Dual choice tests showed that the option of the first host to be accepted by the wasp was random; however, the mean number of parasitized eggs differed significantly when two host species were offered simultaneously to T. cordubensis, always being the host species with heavier eggs the most parasitized.     

Survey for natural enemies of the invasive alien chrysomelid, Diabrotica virgifera virgifera, in Central Europe

The western corn rootworm, Diabrotica virgifera virgifera LeConte(Coleoptera: Chrysomelidae), is the mostdestructive pest of maize (Zea mays L.)in North America, and began to successfullyinvade Central Europe in the early 1990's. Thispaper reports a three-year field surveyconducted in Hungary, Yugoslavia, and Croatia,which are currently the focal points ofinvasion, with the aim to determine theoccurrence of indigenous natural enemies ofD. v. virgifera in Europe. A total of9,900 eggs, 550 larvae, 70 pupae and 33,000adults were examined for the occurrence ofparasitoids, nematodes, and fungal pathogens. It can be concluded from the survey resultsthat effective indigenous natural enemies arenot attacking any of the life stages of D.v. virgifera in Europe. The exception is theoccurrence of the fungi Beauveriabassiana (Bals.) Vuill. (Mitosporic fungi;formerly Deuteromyces) and Metarhiziumanisopliae (Metsch.) Sorok (Mitosporic fungi)attacking adults of D. v. virgifera at anextremely low level (< 1%). However no otherentomopathogenic fungal pathogens,entomopathogenic nematodes, or parasitoids werefound on eggs, larvae, pupae or adults. Whileseveral natural enemies in North and CentralAmerica are known to attack D. v.virgifera, it is apparent that indigenousnatural enemies in Europe have not adapted tothe high population density of the alieninvasive species D. v. virgifera. Classical biological control may provide anopportunity to reconstruct the natural enemycomplex of an invading alien pest, and itsapplication to manage D. v. virgiferapopulations in Europe should be considered.     

Predation by Podisus maculiventris (Hemiptera: Pentatomidae) on Plutella xylostella (Lepidoptera: Plutellidae) larvae parasitized by Cotesia plutellae (Hymenoptera: Braconidae) and its impact on cabbage

Biological control offers potentially effective suppression of the diamondback moth (DBM), Plutella xylostella, a serious pest of Brassica crops. Little is known of whether multiple natural enemies have additive, antagonistic, or synergistic effects on DBM populations. No-choice and choice tests were conducted to assess predation by Podisus maculiventris on DBM larvae parasitized by Cotesia plutellae and unparasitized larvae. In no-choice tests, P. maculiventris preyed on greater numbers of parasitized than unparasitized larvae and greater numbers of young larvae than old larvae. In choice tests with early third instar DBM, there was no difference in predation between parasitized or unparasitized larvae. However, in choice tests with older prey, P. maculiventris preyed on more parasitized than unparasitized larvae. Two field studies were conducted to test if this predator and parasitoid have additive, antagonistic or synergistic effects on DBM populations and plant damage in cabbage (Brassica oleracea var. capitata). In 2002, DBM populations were significantly lower in the presence of C. plutellae but not in the presence of P. maculiventris. There was not a significant interaction between the natural enemies. Plant damage was reduced only with C. plutellae. In 2003, DBM populations were significantly lower in the presence of C. plutellae and P. maculiventris, although the combination of natural enemies did not lead to a non-additive interaction. Plant damage was unaffected by the presence of either natural enemy. Because of its greater predation on parasitized larvae, P. maculiventris could be an intraguild predator of C. plutellae. Yet, their overall combined effect in the field was additive rather than antagonistic.     

Production of Anagrus epos Girault (Hymenoptera: Mymaridae) on Homalodisca vitripennis (Germar) (Hemiptera: Cicadellidae) eggs

Anagrus epos Girault (Hymenoptera: Mymaridae) is a natural enemy candidate for a classical biological control program targeting the glassy-winged sharpshooter (GWSS), Homalodisca vitripennis (Germar) (Hemiptera: Cicadellidae), in California. Little is known about the biology or ecology of A. epos when it utilizes GWSS eggs as a host. Here, we report the results of laboratory studies that describe the host age preference for oviposition, longevity of A. epos adults provided with different food sources, and developmental rates at six different constant temperature regimes. Anagrus epos is a gregarious parasitoid in GWSS eggs with up to 14 adults emerging from each GWSS egg. In choice and no-choice tests for oviposition, A. epos females successfully parasitized all developmental ages of GWSS eggs (1–8 days old). In choice tests, parasitism rates were significantly higher in 1-, 3-, 4-, and 5-day-old GWSS eggs than in 2-, 6-, 7-, and 8-day-old eggs. If provided with honey and water, honey only, water only, or no food or water, A. epos females lived on average 8.2, 4.7, 2.6, and 1.6 days, respectively. Anagrus epos required 294.1 degree-days above a lower temperature threshold of 12.4 °C to develop from egg to adult (eclosion). Our results provide baseline information useful in the development of an efficient parasitoid mass rearing program for A. epos release and evaluation in California.     

Risk to non-target plants from Charidotis auroguttata (Chrysomelidae: Coleoptera), a potential biocontrol agent for cat’s claw creeper Macfadyena unguis-cati (Bignoniaceae) in Australia

The potential of the leaf beetle Charidotis auroguttata as a biocontrol agent for cat’s claw creeper Macfadyena unguis-cati (Bignoniaceae), an environmental weed in Australia, and risk to non-target plants was evaluated under quarantine conditions. In no-choice tests, C. auroguttata adults and larvae fed on many plant species across different families, but egg to adult development occurred only on the target weed. However, when neonate larvae from the target weed were transferred onto Myoporum boninense australe (Myoporaceae), a non-target native plant, 11.7% completed development, as compared to 95% of larvae that completed development on the target weed. Larval development on this non-target species also took twice as long as on the target weed. No larvae completed development on other test plants. In choice tests, leaf area consumption by adults and larvae was significantly more on the target weed than on other plants, and oviposition occurred only on the target weed. In the no-choice demography trials, adults laid eggs from the second week after emergence on the target weed, with an average of 0.286 eggs/female/day, resulting in an 18-fold increase in the adult population over 16 weeks. On My. boninense australe adult survival remained high, but oviposition commenced only from the 10th week after emergence with an average of 0.023 eggs/female/day, and none of the eggs developed into adults. In the choice demography trials, oviposition on the target weed was evident from the fourth week onwards, while on the non-target plant oviposition commenced only from the 14th week. Only 10% of total adults and 11.3% of total eggs were found on the non-target plant, and none of these eggs developed into adults. Although the biocontrol agent can ‘spill-over’ from the target weed to the non-target native plant and cause adult feeding damage, the non-target plant could not sustain a viable insect population on its own. This agent was not approved for field release in Australia due to perceived risk to non-target species.     

Host age choice for parasitism by Lysiphlebia mirzai and its effect on the development and reproduction of brown citrus aphid

The effects of host age andparasitism by Lysiphlebia mirzaiShuja-Uddin (Hymenoptera: Braconidae) on the development, survivorship,reproduction, and life table parameters ofbrown citrus aphid, Toxoptera citricida(Kirkaldy) (Homoptera:Aphididae) were evaluated at 25 ± 1 °C, 80 ± 5% RH, and a photoperiodof 14:10 (L:D) h. The results showed that theduration from initial parasitization tomummification was independent of the aphid ageat the time of parasitization with a mean of6.0 d. Aphids parasitized at the 1st and 2ndinstar stages and 17% of those parasitized atthe 3rd instar stage failed to reach adulthood,while the rest of the aphids parasitized at the3rd and 4th instar stages and as adults reachedmaturity and produced a number of offspringthat increased with age at parasitization. Thelongevity and fecundity of parasitized aphidswere strongly correlated with the aphid age atparasitization. Aphids parasitized at the 1stand 2nd instar stages did not contribute to thepopulation growth of the aphids. The netreproductive rate (R_o) of the aphidsparasitized at the 3rd and 4th instar andadult stages were only 9, 20 and 43% of thatfor nonparasitized aphids, respectively. Hostage preference experiments involving all fiveaphid stages showed L. mirzai couldattack and parasitize all life stages of browncitrus aphid. Parasitism was highest in the2nd instar in a no-choice test, while in achoice test parasitism in the 2nd and 3rdinstars was similar. A relatively high rate ofparasitism was also observed in the 1st instarnymphs in both choice and no-choice tests. Theresults suggest that L. mirzai haspotential as a biocontrol agent of the browncitrus aphid and warrants a furtherevaluation.     

Distribution, host specificity, and overwintering of Celatoria bosqi Blanchard (Diptera: Tachinidae), a South American parasitoid of Diabrotica spp. (Coleoptera: Chrysomelidae: Galerucinae)

The genus Diabrotica (Coleoptera: Chrysomelidae) includes a great number of pest species, including some of the most important crops pests of the Americas. However, only five parasitoid species have been recorded for it. The parasitoid Celatoria bosqi Blanchard was the first parasitoid described from Diabrotica spp. in South America, where substantial parasitism has been observed. C. bosqi has been collected almost throughout the South American distribution of its main host, Diabrotica speciosa (Germar), in an area that includes temperate and tropical lowlands, and semiarid to humid highlands. Three Diabrotica species were found to host the parasitoid, D. speciosa (Germar), Hystiopsis sp., and Diabrotica viridula (F.), with a total parasitism of 2.60, 5.55, and <0.02%, respectively. Laboratory experiments with field beetles and puparia, reared in the laboratory, indicate that C. bosqi overwinters obligatorily in overwintering adult host beetles, remaining quiescent in its live host below developmental temperatures. Based on the known climatic range of C. bosqi, and its requirement of adult overwintering hosts, a potential distribution in North America is projected.     

Relative host plant species use by the lantana biological control agent Aconophora compressa (Membracidae) across its native and introduced ranges

Aconophora compressa Walker (Hemiptera: Membracidae) was released in 1995 against the weed lantana in Australia, and is now found on multiple host plant species. The intensity and regularity at which A. compressa uses different host species was quantified in its introduced Australian range and also its native Mexican range. In Australia, host plants fell into three statistically defined categories, as indicated by the relative rates and intensities at which they were used in the field. Fiddlewood (Citharexylum spinosum L.: Verbenaceae) was used much more regularly and at higher densities than any other host sampled, and alone made up the first group. The second group, lantana (Lantana camara L.: Verbenaceae; pink variety) and geisha girl (Duranta erecta L.: Verbenaceae), were used less regularly and at much lower densities than fiddlewood. The third group, Sheena’s gold (another variety of D. erecta), jacaranda (Jacaranda mimosifolia D. Don: Bignoniaceae) and myoporum (Myoporum acuminatum R. Br.: Myoporaceae), were used infrequently and at even lower densities. In Mexico, the insect was found at relatively low densities on all hosts relative to those in Australia. Densities were highest on L. urticifolia, D. erecta and Tecoma stans (L.) Juss. ex Kunth (Bignoniaceae), which were used at similar rates to one another. It was found also on a few other verbenaceous and non-verbenaceous host species but at even lower densities. The relative rate at which Citharexylum spp. and L. urticifolia were used could not be assessed in Mexico because A. compressa was found on only one plant of each species in areas where these host species co-occurred. The low rate at which A. compressa occurred on fiddlewood in Mexico is likely to be an artefact of the short-term nature of the surveys or differences in the suites of Citharexylum and Lantana species available there. These results provide further incentive to insist on structured and quantified surveys of non-target host use in the native range of potential biological control agents prior to host testing studies in quarantine.     

Comparative susceptibility of larval instars and pupae of the western corn rootworm to infection by three entomopathogenic nematodes

As a first step towards the development of an ecologically rational control strategy against western corn rootworm (WCR; Diabrotica virgifera virgifera LeConte, Coleoptera: Chrysomelidae) in Europe, we compared the susceptibility of the soil living larvae and pupae of this maize pest to infection by three entomopathogenic nematode (EPN) species. In laboratory assays using sand-filled trays, Heterorhabditis bacteriophora Poinar and H. megidis Poinar, Jackson & Klein (both Rhabditida: Heterorhabditidae) caused comparable mortality among all three larval instars and pupae of D. v. virgifera. In soil-filled trays, H. bacteriophora was slightly more effective against third larval instars and pupae, and H. megidis against third larval instars, compared to other developmental stages. In both sand and soil, Steinernema feltiae (Filipjev) (Rh.: Steinernematidae) was least effective against second instars. In conclusion, all larval instars of D. v. virgifera show susceptibility to infection by all three nematodes tested. It is predicted that early application against young larval instars would be most effective at preventing root feeding damage by D. v. virgifera. Applications of nematodes just before or during the time period when third instars are predominant in the field are likely to increase control efficacy. According to our laboratory assays, H. bacteriophora and H. megidis appear to be the most promising candidates for testing in the field. I. Hiltpold similarly contributed to this paper as the first author.     

The suitability of non-target native mangroves for the survival and development of the lantana bug <Emphasis Type="Italic">Aconophora compressa</Emphasis>, an introduced weed biological control agent

Aconophora compressa (Hemiptera: Membracidae), a biological control agent introduced against the weed Lantana camara (Verbenaceae) in Australia, has since been observed on several non-target plant species, including native mangrove Avicennia marina (Acanthaceae). In this study we evaluated the suitability of two native mangroves, A. marina and Aegiceras corniculatum (Myrsinaceae), for the survival and development of A. compressa through no-choice field cage studies. The longevity of females was significantly higher on L. camara (57.7 ± 3.8 days) than on A. marina (43.3 ± 3.3 days) and A. corniculatum (45.7 ± 3.8 days). The proportion of females laying eggs was highest on L. camara (72%) followed by A. marina (36%) and A. corniculatum (17%). More egg batches per female were laid on L. camara than on A. marina and A. corniculatum. Though more nymphs per shoot emerged on L. camara (29.9 ± 2.8) than on A. marina (13 ± 4.8) and A. corniculatum (10 ± 5.3), the number of nymphs that developed through to adults was not significantly different. The duration of nymphal development was longer on A. marina (67 ± 5.8 days) than on L. camara (48 ± 4 days) and A. corniculatum (43 ± 4.6 days). The results, which are in contrast to those from previous glasshouse and quarantine trials, provide evidence that A. compressa adults can survive, lay eggs and complete nymphal development on the two non-target native mangroves in the field under no-choice condition.     

Host Preferences of Trichogramma pretiosum and the Influence of Prior Ovipositional Experience on the Parasitism of Plutella xylostella and Pseudoplusia includens Eggs

Trichogramma wasps are generalist egg parasitoids used in biological control efforts. In a multi host situation they may preferentially parasitize a non-target host species to the detriment of the control program. Plutella xylostella Linnaeus (Lepidoptera: Plutellidae) is a very serious pest of cabbage, but is only one in a number of species in the ‘cabbageworm’ complex. We investigated the host preferences of Trichogramma pretiosum Riley (Hymenoptera: Chalcidoidea) when offered the eggs of Plutella xylostella and the eggs of Pseudoplusia includens Hübner (Lepidoptera: Noctuidae), another species in the ‘cabbageworm’ complex. Trichogramma pretiosum reared on the eggs of the factitious host Ephestia kuehniella Zeller (Lepidoptera: Pyralidae) parasitized both Plutella xylostella and Pseudoplusia includens eggs under laboratory conditions. For both choice and no-choice experiments, T. pretiosum parasitized significantly more P. xylostella eggs than P. includens eggs. Prior ovipositional experience with one or other of the two host species had no effect on the subsequent parasitism levels of the two host species. The preference for P. xylostella eggs was also not affected by this prior ovipositional experience.     

Natural occurrence of insect pathogenic fungi and insect parasitic nematodes in <Emphasis Type="Italic">Diabrotica virgifera virgifera</Emphasis> populations

The Western Corn Rootworm D. virgifera virgifera Le Conte (Coleoptera: Chrysomelidae), a serious pest of maize, has been recently introduced into Europe. Several approaches for its control are presently under investigation including microbial agents. In order to get information on the role of naturally occurring pathogens in the regulation of Diabrotica populations, we started an investigation in established populations in Hungary, Romania, Serbia, Austria, and Italy in 2005 and 2006. In infested maize fields in Hungary, plants and their root systems were grubbed out and larvae and pupae were collected. Adult D. v. virgifera were collected in Hungary, Austria, Romania, Serbia and Italy. Additionally, the occurrence of entomopathogenic fungi in soils of maize fields was determined using Galleria mellonella and Tenebrio molitor larvae as bait insects. The density of entomopathogenic fungi was obtained by plating soil suspension on selective medium. Metarhizium anisopliae and Beauveria spp. infections were found in 1.4% of field collected larvae, 0.2% of field collected pupae and 0.05% of field collected adults. Whereas natural infections of D. v. virgifera were rarely found, a high density of insect pathogenic fungi was recorded in Hungarian soils. M. anisopliae could be detected in every maize field either using the “bait method” or a “selective medium” method. This is the first report of a natural occurrence of entomoparasitic nematodes (Heterorhabditis sp., Steinernema sp.) in Diabrotica v. virgifera in Europe.     

Intra- and interspecific competition by Fopius arisanus and Diachasmimorpha tryoni (Hymenoptera: Braconidae), parasitoids of tephritid fruit flies

Fopius arisanus (Sonan) and Diachasmimorpha tryoni (Cameron) are two important solitary endoparasitoids of tephritid fruit flies. The former species attacks host eggs while the latter attacks host larvae, and both species emerge as adults from the host puparium. This study investigated intrinsic competition between these two parasitoids, as well as aspects of intraspecific competition within each species in the Mediterranean fruit fly, Ceratitis capitata (Wiedemann). Parasitization by F. arisanus resulted in direct mortality of host eggs and prolonged development of host eggs and larvae. Superparasitism by F. arisanus was uncommon when mean parasitism per host patch was <50%, but increased with rising rates of parasitism. Superparasitism by D. tryoni was more common. In superparasitized hosts, supernumerary individuals of F. arisanus were killed through physiological suppression, while supernumerary larvae of D. tryoni were killed mainly through physical attack. In multiparasitized hosts, dissections showed that 81.6% of D. tryoni eggs in the presence of F. arisanus larvae died within 3 days, indicating physiological inhibition of egg hatch. Rearing results further showed that F. arisanus won almost all competitions against D. tryoni. The ratio of D. tryoni stings to ovipositions was lower in hosts not previously parasitized by F. arisanus than in parasitized hosts, suggesting that D. tryoni can discriminate against parasitized hosts. The mechanism that F. arisanus employs to eliminate D. tryoni is similar to that it uses against all other larval fruit fly parasitoids so far reported. The results are discussed in relation to the competitive superiority of early acting species in fruit fly parasitoids, and to a possible competitive-mediated mechanism underlying host shift by D. tryoni to attack non-target flies following the successful introduction of F. arisanus in Hawaii.     

Open field host selection and behavior by tamarisk beetles (Diorhabda spp.) (Coleoptera: Chrysomelidae) in biological control of exotic saltcedars (Tamarix spp.) and risks to non-target athel (T. aphylla) and native Frankenia spp.

Biological control of invasive saltcedars (Tamarix spp.) in the western U.S. by exotic tamarisk leaf beetles, Diorhabda spp., first released in 2001 after 15 years of development, has been successful. In Texas, beetles from Crete, Greece were first released in 2004 and are providing control. However, adults alight, feed and oviposit on athel (Tamarix aphylla), an evergreen tree used for shade and as a windbreak in the southwestern U.S. and México, and occasionally feed on native Frankenia spp. plants. The ability of tamarisk beetles to establish on these potential field hosts was investigated in the field. In no-choice tests in bagged branches, beetle species from Crete and Sfax, Tunisia produced 30–45% as many egg masses and 40–60% as many larvae on athel as on saltcedar. In uncaged choice tests in south Texas, adult, egg mass and larval densities were 10-fold higher on saltcedar than on adjacent athel trees after 2 weeks, and damage by the beetles was 2- to 10-fold greater on saltcedar. At a site near Big Spring, in west-central Texas, adults, egg masses and 1st and 2nd instar larvae were 2- to 8-fold more abundant on saltcedar than on athel planted within a mature saltcedar stand being defoliated by Crete beetles, and beetles were 200-fold or less abundant or not found at all on Frankenia. At a site near Lovelock, Nevada, damage by beetles of a species collected from Fukang, China was 12–78% higher on saltcedar than on athel planted among mature saltcedar trees undergoing defoliation. The results demonstrate that 50–90% reduced oviposition on athel and beetle dispersal patterns within resident saltcedar limit the ability of Diorhabda spp. to establish populations and have impact on athel in the field.     

Pollen beetle (Meligethes aeneus) oviposition and feeding preference on different host plant species

The oviposition and feeding preferences of the pollen beetle, Meligethes aeneus, were determined in choice and no-choice tests in field, semi-field and greenhouse trials. Plant species used were Brassica napus, B. campestris, B. juncea, B. nigra, B. carinata, Sinapis alba and Crambe abyssinica. With respect to number of eggs laid, S. alba and C. abyssinica were inferior to the other species. Pollen beetles laid fewer eggs on B. nigra than on the other Brassica spp. in no-choice tests, however this difference was partly due to fewer eggs laid per bud rather than fewer buds used for oviposition. Most eggs, for all plant species, were deposited in buds sized 2–3 mm. Feeding damage on all plant species was relatively similar. Pollen beetles seem to have a wider host range for feeding than for oviposition. There was good agreement in plant species ranking as oviposition hosts between the field, semi-field and greenhouse trials.     

Supplementary host specificity testing of the sawfly <Emphasis Type="Italic">Heteroperryia hubrichi</Emphasis>, a candidate for classical biological control of Brazilian peppertree, <Emphasis Type="Italic">Schinus terebinthifolius</Emphasis>, in the USA

Supplementary host specificity tests were conducted with the defoliating sawfly Heteroperreyia hubrichi (Hymenoptera: Pergidae), a candidate for classical biological control of Brazilian peppertree, Schinus terebinthifolius (Sapindales: Anacardiaceae), in the United States. These tests were conducted as part of the environmental assessment required by the National Environmental Policy Act (NEPA). The suitability of the federally listed endangered native plant Rhus michauxii (Anacardiaceae) and the economically important Litchi chinensis (Sapindaceae) as potential host plants for H. hubrichi was evaluated in a series of no-choice larval development tests conducted in a Florida quarantine laboratory. Neonate larvae transferred to individual test plants failed to develop to the pupal stage on R. michauxii or L. sinensis, whereas Brazilian peppertree supported development of the sawfly to pupation. The results of the additional host specificity tests indicated these critical non-target plants are not at risk from attack by H. hubrichi if it were released in Florida, USA.     

Potential for habituation to a neem-based feeding deterrent in Japanese beetles, Popillia japonica

We tested the potential for the Japanese beetle, Popillia japonica Newman, to habituate to a neem-based feeding deterrent applied to foliage of linden, Tilia cordata L., a preferred host for the adults. Female beetles' consumption of control foliage versus foliage treated with either a low or high rate of neem insecticide, corresponding to 9 or 39 pm azadirachtin, respectively, was tested in a series of 4-h choice or no-choice tests over four successive days. In another experiment, females were conditioned for 22 h with either control foliage, leaves treated with the low rate, or a mixture of both treated and untreated leaves. Deterrence of either the low or high rate of neem to these beetles was then evaluated in choice tests with control foliage, as before. In choice tests, mean consumption of control foliage was always greater than for treated foliage, regardless of rate. There was, however, proportionately more feeding on foliage treated with the high rate upon successive exposures. In no-choice tests, beetles initially deterred by the low rate were not significantly deterred by that rate by the third and fourth days of the experiment. Finally, beetles conditioned by exposure to leaves treated with the low rate were not deterred by that rate in a subsequent choice test, although they were deterred by the higher rate. Despite these trends, we suggest that Japanese beetles' polyphagy and mobility probably would reduce the likelihood for habituation to neem-based feeding deterrents in the field.