Insect hemocytes and their role in immunity

The innate immune system of insects is divided into humoral and cellular defense responses. Humoral defenses include antimicrobial peptides, the cascades that regulate coagulation and melanization of hemolymph, and the production of reactive intermediates of oxygen and nitrogen. Cellular defenses refer to hemocyte-mediated responses like phagocytosis and encapsulation. In this review, we discuss the cellular immune responses of insects with emphasis on studies in Lepidoptera and Diptera. Insect hemocytes originate from mesodermally derived stem cells that differentiate into specific lineages identified by morphology, function, and molecular markers. In Lepidoptera, most cellular defense responses involve granular cells and plasmatocytes, whereas in Drosophila they involve primarily plasmatocytes and lamellocytes. Insect hemocytes recognize a variety of foreign targets as well as alterations to self. Both humoral and cell surface receptors are involved in these recognition events. Once a target is recognized as foreign, hemocyte-mediated defense responses are regulated by signaling factors and effector molecules that control cell adhesion and cytotoxicity. Several lines of evidence indicate that humoral and cellular defense responses are well-coordinated with one another. Cross-talk between the immune and nervous system may also play a role in regulating inflammation-like responses in insects during infection.     

Innate immunity and its evasion and suppression by hymenopteran endoparasitoids

Recent studies suggest that insects use pattern recognition molecules to distinguish prokaryotic pathogens and fungi from "self" structures. Less understood is how the innate immune system of insects recognizes endoparasitic Hymenoptera and other eukaryotic invaders as foreign. Here we discuss candidate recognition factors and the strategies used by parasitoids to overcome host defense responses. We suggest that host-parasitoid systems are important experimental models for studying how the innate immune system of insects recognizes foreign invaders that are phylogenetically more closely related to their hosts. The strategies used by parasitoids suggest that insects may employ "hidden-self" recognition molecules for attacking foreign objects intruding the open circulatory system. BioEssays 23:344-351, 2001.     

Suppressing resistance to Bt cotton with sterile insect releases

Genetically engineered crops that produce insecticidal toxins from Bacillus thuringiensis (Bt) are grown widely for pest control. However, insect adaptation can reduce the toxins' efficacy. The predominant strategy for delaying pest resistance to Bt crops requires refuges of non-Bt host plants to provide susceptible insects to mate with resistant insects. Variable farmer compliance is one of the limitations of this approach. Here we report the benefits of an alternative strategy where sterile insects are released to mate with resistant insects and refuges are scarce or absent. Computer simulations show that this approach works in principle against pests with recessive or dominant inheritance of resistance. During a large-scale, four-year field deployment of this strategy in Arizona, resistance of pink bollworm (Pectinophora gossypiella) to Bt cotton did not increase. A multitactic eradication program that included the release of sterile moths reduced pink bollworm abundance by >99%, while eliminating insecticide sprays against this key invasive pest.     

The influence of host and non‐host companion plants on the behaviour of pest insects in field crops

Companion plants grown as ‘trap crops’ or ‘intercrops’ can be used to reduce insect infestations in field crops. The ways in which such reductions are achieved are being described currently using either a chemical approach, based on the ‘push‐pull strategy’, or a biological approach, based on the ‘appropriate/inappropriate landing theory’. The chemical approach suggests that insect numbers are reduced by chemicals from the intercrop ‘repelling’ insects from the main crop, and by chemicals from the trap‐crop ‘attracting’ insects away from the main crop. This approach is based on the assumptions that (1) plants release detectable amounts of volatile chemicals, and (2) insects ‘respond’ while still some distance away from the emitting plant. We discuss whether the above assumptions can be justified using the ‘appropriate/inappropriate landing theory’. Our tenet is that specialist insects respond only to the volatile chemicals released by their host plants and that these are released in such small quantities that, even with a heightened response to such chemicals, specialist insects can only detect them when a few metres from the emitting plant. We can find no robust evidence in the literature that plant chemicals ‘attract’ insects from more than 5 m and believe that ‘trap crops’ function simply as ‘interception barriers’. We can also find no evidence that insects are ‘repelled’ from landing on non‐host plants. Instead, we believe that ‘intercrops’ disrupt host‐plant finding by providing insects with a choice of host (appropriate) and non‐host (inappropriate) plant leaves on which to land, as our research has shown that, for intercropping to be effective, insects must land on the non‐host plants. Work is needed to determine whether non‐host plants are repellent (chemical approach) or ‘non‐stimulating’ (biological approach) to insects.     

Cellular defence reactions and their depression in insects]

In insects the main cellular defence reactions are phagocytosis and encapsulation of foreign bodies. Free cells of haemolymph called haemocytes are the effectors of these reactions. They are achieved under the control of humoral factors of the plasma or of the serum. Humoral factors are able to enhance or to decrease the cellular defence reactions. As in mammals, potential parasites or pathogens need to avoid or to inhibit the defence reactions before developing inside the insect body. As an example of the depression of immunity induced by a parasite we will study the relationships between an insect and a nematobacterial complex.     

昆虫与植物关系的研究进展和前景

本文综述昆虫与植物之间关系的研究概况，包括历史渊源、昆虫选择寄主植物的生理机制，植物对虫害的反应、用抗虫基因在作物中移植以防治害虫和展望。着重叙述昆虫神经中枢对于植物理化特性所产生的感觉内导的综合作用，植物蒙受虫害后的补偿作用及由此诱导所产生的化学防御作用。讨论了以抗虫基因移植于农林作物来防治害虫是否会引起昆虫对这种新育成的植物产生适应或抗性。昆虫与植物之间的关系是一个重要的科研领域，对其发展前景     

The survival of ingested Serratia marcescens in houseflies (Musca domestica L.) after electrocution with electric fly killers

Electric fly killers (EFKs) are commonly used to control flying insects that enter food establishments. For establishment of the incidence of pathogen-bearing insects in food establishments, insect samples obtained from EFK trays could be used. The principal difficulty with this approach is that the survival time of microorganisms on or within insect corpses after electrocution is unknown. This study determined the survival of Serratia marcescens (as a representative of the enteric bacteria) within houseflies following their electrocution by a commercial EFK. S. marcescens was successfully ingested by houseflies and survived on and within the corpses after electrocution for up to 5 weeks. Maximal levels of bacteria were recovered 24 h postelectrocution. The study also demonstrates the ability of ingested S. marcescens to out-compete resident microbial flora within houseflies. The findings are intended to pave the way for further research to determine the incidence of pathogen-laden flying insects in food establishments.     

Mechanisms underlying insect freeze tolerance

Freeze tolerance – the ability to survive internal ice formation – has evolved repeatedly in insects, facilitating survival in environments with low temperatures and/or high risk of freezing. Surviving internal ice formation poses several challenges because freezing can cause cellular dehydration and mechanical damage, and restricts the opportunity to metabolise and respond to environmental challenges. While freeze‐tolerant insects accumulate many potentially protective molecules, there is no apparent ‘magic bullet’ – a molecule or class of molecules that appears to be necessary or sufficient to support this cold‐tolerance strategy. In addition, the mechanisms underlying freeze tolerance have been minimally explored. Herein, we frame freeze tolerance as the ability to survive a process: freeze‐tolerant insects must withstand the challenges associated with cooling (low temperatures), freezing (internal ice formation), and thawing. To do so, we hypothesise that freeze‐tolerant insects control the quality and quantity of ice, prevent or repair damage to cells and macromolecules, manage biochemical processes while frozen/thawing, and restore physiological processes post‐thaw. Many of the molecules that can facilitate freeze tolerance are also accumulated by other cold‐ and desiccation‐tolerant insects. We suggest that, when freezing offered a physiological advantage, freeze tolerance evolved in insects that were already adapted to low temperatures or desiccation, or in insects that could withstand small amounts of internal ice formation. Although freeze tolerance is a complex cold‐tolerance strategy that has evolved multiple times, we suggest that a process‐focused approach (in combination with appropriate techniques and model organisms) will facilitate hypothesis‐driven research to understand better how insects survive internal ice formation.     

Knocking down the sex peptide receptor by dsRNA feeding results in reduced oviposition rate in olive fruit flies

Insect pests can cause crop damage in yield or quality, resulting in profit losses for farmers. The primary approach to control them is still the use of chemical pesticides resulting in significant hazards to the environment and human health. Biological control and the sterile insect technique are alternative strategies to improve agriculture protection. However, both strategies have significant limitations. A newly introduced approach that could be both effective and species-specific is the RNA interference mechanism. One key point for the success of this strategy is the delivery method of double-strand RNA (dsRNA) to the insects. A method of dsRNA delivery to insects with potential use in the field is the oral delivery, feeding the insects engineered microorganisms that produce dsRNA. Here, we present the first protocol for dsRNA feeding using modified bacteria, in the olive fruit fly, the most important insect pest of cultivated olives. We chose to target the sex peptide receptor gene. The sex peptide receptor interacts with the sex peptide, a peptide that is responsible for the postmating behavior in the model organism, Drosophila melanogaster. Feeding the female olive fruit fly with bacteria that produced dsRNA for the sex peptide receptor gene resulted in the development of female insects with significantly lower oviposition rates. Administration of dsRNA producing bacteria in insect diet against target genes that lead to genetic sexing or female-specific lethality could be added in the armory of control methods.     

Identification of insect pests as food foreign materials using DNA barcodes

Insect foreign materials in food are of great economic and hygienic significance. However, identifying these species with any certainty requires an expert taxonomist and can be a time consuming process. Furthermore, insects are found as body parts or they are immature specimens that cannot be identified by conventional means. For these reasons, a reference database and efficient means of identification by non‐specialists are necessary to control insect pests. In this study, we chose 15 important insect pest species, because they had a higher probability of being included in human foods. We tested the utility of the cytochrome c oxidase I (COI) DNA barcodes for the identification. A 658‐bp fragment of the COI gene was sequenced, aligned, and a sequence data bank was constructed. As a result, the COI barcode sequence was suitable for identifying insect pests as food foreign materials. Photographs of morphological key characters by stereoscopic microscope and a pictorial key of the species are provided.     

Parasitoid Encapsulation as a Defense Mechanism in the Coccoidea (Homoptera) and Its Importance in Biological Control

Encapsulation is a common defense mechanism exerted by a host insect in response to invasion by a metazoan parasitoid or other foreign organisms. In the process of encapsulation, the host forms a capsule around the parasitoid egg or larva, which is usually composed of host blood cells and the pigment melanin. The capsule may kill the parasitoid and thus prevent successful parasitism. Encapsulation may adversely affect the degree of biological control effected by parasitoids as it may either prevent the establishment of exotic parasitoids in new regions or reduce parasitoid efficacy. A high incidence of encapsulation may also cause difficulties in mass rearing of parasitoids. In the Coccoidea (Homoptera), parasitoid encapsulation has so far been recorded in three families: Coccidae (soft scale insects), Diaspididae (armored scale insects), and Pseudococcidae (mealybugs). Important factors that affect the frequency of parasitoid encapsulation in the Coccoidea include: Host and parasitoid species, the host's physiological age and physiological condition, the host origin (or strain), superparasitism, the rearing and/or ambient temperature, and the host plant. The effects of these factors on the incidence of parasitoid encapsulation in the Coccidae, Diaspididae, and Pseudococcidae are described.     

Insect population control using female specific pro-drug activation

A system for population control of insects is proposed. It is based on transgenic insects expressing an enzyme which converts an inactive pro-drug into an active, toxic form. A model system is presented which relies on transposon-mediated integration of a bacterial cytosine deaminase (CD) gene into the genome of Drosophila melanogaster. We demonstrate female-specific sterility and transgene-dependent lethality when flies carrying the CD gene under a Drosophila female-specific promoter/enhancer are treated with 5-Fluorocytosine, a low-toxicity nucleoside analogue which is converted to toxic 5-Fluorouracil by the enzyme. The approach can be used with existing pro-insecticides and appropriate converting enzymes in combination with established mass rearing technology, for targeted, environmentally acceptable control of insects of economic and public health importance.     

Using optimality models to improve the efficacy of parasitoids in biological control programmes

Biological control of insect pests relies on the ability of natural enemies to limit pest populations. The behaviours expressed by natural enemies against their prey or hosts are modulated by a number of factors and a better understanding of these factors is key to obtaining more efficacious pest control. We propose here that optimality models based upon a behavioural ecology approach can provide a framework that should enable optimisation of biological control practices. We limit our discussion to parasitoid natural enemies and review the factors known to influence the behaviour of these insects. The most important areas that have been studied extensively in the behavioural ecology of insect parasitoids are addressed here: (1) residence time in a host patch, (2) clutch size, (3) sex ratio, (4) host and patch marking, and (5) diet choice. We discuss the implications of the incorporation of these optimality models into efficacious biological control practices and suggest areas where a better knowledge of the behavioural ecology of these insects could improve the efficacy of parasitoid‐based pest control.     

Recognition in ants: social origin matters

The ability of group members to discriminate against foreigners is a keystone in the evolution of sociality. In social insects, colony social structure (number of queens) is generally thought to influence abilities of resident workers to discriminate between nestmates and non-nestmates. However, whether social origin of introduced individuals has an effect on their acceptance in conspecific colonies remains poorly explored. Using egg-acceptance bioassays, we tested the influence of social origin of queen-laid eggs on their acceptance by foreign workers in the ant Formica selysi. We showed that workers from both single- and multiple-queen colonies discriminated against foreign eggs from single-queen colonies, whereas they surprisingly accepted foreign eggs from multiple-queen colonies. Chemical analyses then demonstrated that social origins of eggs and workers could be discriminated on the basis of their chemical profiles, a signal generally involved in nestmate discrimination. These findings provide the first evidence in social insects that social origins of eggs interfere with nestmate discrimination and are encoded by chemical signatures.     

Accelerated development of quarantine treatments for insects on poor hosts.

The probit 9 standard for quarantine treatment efficacy (99.9968% mortality) was originally recommended for tropical fruits heavily infested with fruit flies and it centers on high mortality to achieve quarantine security. This standard may be too stringent for quarantine pests in commodities that are rarely infested or are poor hosts, The alternative treatment efficacy approach measures risk as the probability of a mating pair, gravid female, or parthenogenic individual surviving in a shipment. This will be a function of many factors including infestation rate and shipment volume. Applying the risk-based alternative treatment efficacy approach to pests on rarely infested or poor hosts will lower the number of required test insects needed for developing quarantine treatments; hence data for a quarantine treatment could be generated by testing 10,000 or fewer insects with no survivors, compared with 90,000-100,000 insects to demonstrate the traditional probit 9 efficacy. Several commodity/quarantine pest systems where this approach could be applied are discussed. This approach would save time and resources, and help farmers export their crop on a more-timely basis.     

The Survival of Ingested <Emphasis Type="Italic">Serratia marcescens</Emphasis> in Houseflies (<Emphasis Type="Italic">Musca domestica</Emphasis> L.) After Electrocution with Electric Fly Killers

Electric fly killers (EFKs) are commonly used to control flying insects that enter food establishments. For establishment of the incidence of pathogen-bearing insects in food establishments, insect samples obtained from EFK trays could be used. The principal difficulty with this approach is that the survival time of microorganisms on or within insect corpses after electrocution is unknown. This study determined the survival of Serratia marcescens (as a representative of the enteric bacteria) within houseflies following their electrocution by a commercial EFK. S. marcescens was successfully ingested by houseflies and survived on and within the corpses after electrocution for up to 5 weeks. Maximal levels of bacteria were recovered 24 h postelectrocution. The study also demonstrates the ability of ingested S. marcescens to out-compete resident microbial flora within houseflies. The findings are intended to pave the way for further research to determine the incidence of pathogen-laden flying insects in food establishments. Received: 30 April 2002 / Accepted: 3 July 2002     

o-quinone/quinone methide isomerase: a novel enzyme preventing the destruction of self-matter by phenoloxidase-generated quinones during immune response in insects

Melanization and encapsulation of invading foreign organisms observed during the immune response in insects is known to be due to the action of activated phenoloxidase. Phenoloxidase-generated quinones are deposited either directly or after self-polymerization on foreign objects accounting for the observed reactions. Since the reactions of quinones are nonenzymatic, they do not discriminate self from nonself and hence will also destroy self-matter. In this report we present evidence for the presence of a novel quinone/quinone methide isomerase in the hemolymph of Sarcophaga bullata which destroys long-lived quinones and hence acts to protect the self-matter. Quinone methides, formed by the action of this enzyme on physiologically important quinones, being unstable undergo rapid hydration to form nontoxic metabolites.     

双翅目害虫性别分离技术的研究进展及展望

在过去的几十年中,昆虫不育技术(sterile insect technique, SIT)已被用于防治农业害虫和人类健康相关的病媒害虫。相较于传统的农药控制策略,昆虫不育技术具有物种特异性和环境友好型等特点。通过释放不育雄虫的昆虫不育技术的主要障碍是在大规模饲养阶段将雄性与雌性分离,从而提高这些防治方法的成本效率,并防止释放携带和传播疾病的雌性群体。目前大多数针对双翅目害虫的遗传防治策略没有进行性别分离,少数害虫性别分离方法是基于蛹的大小或者雌雄蛹羽化时间差异进行人工识别和机械识别分离。双翅目昆虫性别决定及分化分子机制多种多样,其性别决定主要信号差异巨大,其多种性别决定基因已用于性别分离系统的开发。性比失衡性别分离策略通过破坏性别决定途径关键基因的表达获得雄性偏向后代,雌性条件性致死分离策略利用性别决定关键基因的雌雄选择性剪接差异实现性别分离,这两种性别分离策略目前正在害虫不育防治中接受大规模饲养应用评估,而基于双翅目昆虫雌雄性二态和基因标记发展的可视化性别分离策略也已成功实现多种害虫的性别分离。我们对性比失衡分离策略、雌性条件性致死分离策略和可视化性别分离策略在双翅目害虫中的研究进展进行了综述,重点评估了这些方法在雄虫大规模饲养和释放的应用潜力,以期在更完善的性别分离技术支持下为害虫防治研究取得更多突破性进展。     

Forensic entomology and globalisation

The main aim of Forensic Entomology has always been, and is today, to establish the time of death (P.M.I.: Postmortem Period) or, more exactly, how long a carrion has been exposed in the environment. Most of the invertebrate fauna occurring on corpses consists of insects (mostly Diptera and Coleoptera). They are selectively attracted by the decomposing status of the carrion, and form complex communities or biocenosis within necrophagous or sarcophagous species and their predators, parasites and parasitoids, competing each one another. The rapid and continuos changes of the micro-ecosystem (the body), until its breakdown, does not permit the achievement of a steady state or an equilibrium in the animal communities. These continuous modifications give us the possibility to estimate when (and where) the death has occurred, by the identification of the species feeding on the corpse, the knowledge of their life history, and the length of each stage of their cycle at varying the temperature and the other abiotic factors, external to the carrion ecosystem. The P.M.I. today is still largely based on the tables of faunal succession on human cadavers recognised by Mégin in 1894, with few changes proposed by Authors from other countries. In the last years, however, it happens more and more often, that the natural communities are subverted by the presence of allocton species, which can compete, predate or parasite the most common local sarcophagous insects, modifying, this way, the succession waves and the trophic nets. The immission in the environment of foreign species may be voluntary or casual, but in any case is due to anthropic activities. The voluntary immission happens when some species, employed in the biological struggle against pest or dangerous insects, for pollination of allocton plants, or for other commercial utilities, are beyond man's control and swarm onto the environment; the casual spread is due to the globalisation phenomenon, that distributes the "little organisms" by chance, together with travellers, goods and food items. Together with human migratory flows, raw materials and vegetal foodstuffs travelling from Tropical developing Countries to the North of the world, also many tropical and subtropical insects can be carried out from their original lands. Eurioecious and polyphagous species, and species that evolved mechanisms (as diapausa or hibernation) to get over critical environmental conditions have the highest probability of survival. Saprophagous insects, and flies in particular, evolved such capabilities. The mortality of foreign species due to the difference of temperature and seasonally between tropical and temperate areas was, in the past, the most effective factor limiting the geographic propagation of insects. The ongoing global climate changes induce insect populations, now confined to the tropics, to most likely spread towards middle latitudes, where their specific competitors, predators and parasites, which regulate the population growth, are often absent. The lack of a biological control, the warming up of atmosphere temperature and the fall of the differences among seasons induce a more rapid development and an increased number of generations in new species, that often displace the autocton ones. This phenomenon is much more clear in little and simple ecosystems (such as carrion), mostly occur than in large and complex ecosystems, where many more components, vegetal organisms and phytophagous animals included, are present. To demonstrate how globalisation and climate changes are breaking the geographic barriers, we present some cases in which, during our entomoforensic investigations, performed mainly in North-eastern Italy, Neotropical, African and Asiatic necrophagous flies, beetles and wasp parasitoids have been collected, some of which rare or new for Italy or Paleartic Region. In particular, we report our studies on the american black soldier fly, Hermetia illucens (Diptera: Stratiomyidae), that is showing a heavy competition with the local saprophagous species and is reaching great importance in some man activities and in cattle health.     

Insect quality control: synchronized sex,mating system,and biological rhythm

The sterile insect technique (SIT) is a method of eradicating insects by releasing mass-reared sterilized males into fields to reduce the hatchability of eggs laid by wild females that have mated with the sterile males. SIT requires mass-production of the target insect, and maintenance of the quality of the mass-reared insects. The most important factor is successful mating between wild females and sterile males because SIT depends on their synchronized copulation. Therefore, understanding the mating systems and fertilization processes of target insects is prerequisite. Insect behavior often has circadian rhythms that are controlled by a biological clock. However, very few studies of relationships between sterile insect quality and circadian rhythm have been performed compared with the amount of research on the mating ability of target insects. The timing of male copulation attempts with receptivity of females is key to successful mating between released males and wild females. Therefore, we should focus on the mechanisms controlling the timing of mating in target insects. On the other hand, in biological control projects, precise timing of the release of natural enemies to attack pest species is required because behavior of pests and control agents are affected by their circadian rhythms. Involving both chronobiologists and applied entomologists might produce novel ideas for sterile insect quality control by synchronized sex between mass-reared and wild flies, and for biological control agent quality by matching timing in activity between predator activity and prey behavior. Control of the biological clocks in sterile insects or biological control agents is required for advanced quality control of rearing insects.