Chemical Profile, Antioxidant Properties and Antimicrobial Activities of Malaysian Heterotrigona itama Bee Bread.

The aim of the study was to determine the chemical profile, antioxidant properties and antimicrobial activities of Heterotrigona itama bee bread from Malaysia. The pH, presence of phytochemicals, antioxidant properties, total phenolic content (TPC) and total flavonoid content (TFC), as well as antimicrobial activities, were assessed. Results revealed a decrease in the pH of bee bread water extract (BBW) relative to bee bread ethanolic extract (BBE) and bee bread hot water extract (BBH). Further, alkaloids, flavonoids, phenols, tannins, saponins, terpenoids, resins, glycosides and xanthoproteins were detected in BBW, BBH and BBE. Also, significant decreases in TPC, TFC, DPPH activity and FRAP were detected in BBW relative to BBH and BBE. We detected phenolic acids such as gallic acid, caffeic acid, trans-ferulic acid, trans 3-hydroxycinnamic acid and 2-hydroxycinnamic acid, and flavonoids such as quercetin, kaempferol, apigenin and mangiferin in BBE using high-performance liquid chromatography analysis. The strongest antimicrobial activity was observed in Klebsilla pneumonia (MIC50 1.914 µg/mL), followed by E. coli (MIC50 1.923 µg/mL), Shigella (MIC50 1.813 µg/mL) and Salmonella typhi (MIC50 1.617 µg/mL). Bee bread samples possess antioxidant and antimicrobial properties. Bee bread contains phenolic acids and flavonoids, and could be beneficial in the management and treatment of metabolic diseases.

Some sawfly larvae survive predator-prey interactions with pentatomid Picromerus bidens.

Most Asopinae stinkbugs (Hemiptera: Pentatomidae) prey on other insects, including sawfly larvae (Hymenoptera: Symphyta). Sawfly larvae of the Argidae and Pergidae contain toxic peptides, but whether they are defended against stinkbugs remains poorly studied. A literature survey indicates that no publication is devoted to laboratory tests specifically using these sawflies against stinkbugs. Here, laboratory bioassays were made with the stinkbug Picromerus bidens and four sawfly species at last larval instars: Arge ochropus (Argidae), Arge pagana (also tested at medium instars), Lophyrotoma zonalis (Pergidae), and Allantus rufocinctus (Tenthredinidae). Following 24 h of possible predator-prey interactions, no larvae of A. rufocinctus survived, whereas most or all larvae of the other sawfly species did survive and were still alive 48 h later. When feeding on an argid or pergid larva, the feeding periods lasted on average 6-20 s only, some bugs removing their rostrum and abruptly backing away. Full-grown larvae of A. pagana were attacked less than younger ones. It is likely that the tested Argidae and Pergidae are well defended against P. bidens by potent, internal antifeedants, while defensive body movements combined with a large body size play a secondary role.

Body distribution of toxic peptides in larvae of a pergid and an argid sawfly species.

Larvae of most Pergidae and Argidae (Symphyta: Tenthredinoidea) species contain toxic peptides such as pergidin and lophyrotomin. Here, larval hemolymph and organs of the pergid Lophyrotoma zonalis and the argid Arge pagana were analysed by liquid chromatography-tandem mass spectrometry. The major identified peptides were pergidin and 4-valinepergidin in L. zonalis, whereas pergidin and lophyrotomin in A. pagana. The storage period prior to chemical analysis was longer for the samples of the pergid than the argid species, which influenced peptide concentrations. In both species, however, the peptides occurred in decreasing order of concentration, first in the hemolymph, then in the integument, while minor amounts of the peptides were detected in other organs such as gut and fat body. By separating the cuticle of the pergid from the remaining integument, the peptides were found in equivalent amounts in each of these two body structures. The results suggest that the peptides play an important role in the defence of these sawfly larvae against predators.

Berberis sawfly contains toxic peptides not only at larval stage.

Livestock can die from grazing in areas where larvae of certain Argidae or Pergidae species containing toxic peptides occur in mass. However, it remains unknown whether other stages also contain these compounds. Here, single specimens of larvae, prepupae, and adults of Arge berberidis, plus samples of its cocoons and larval feces, were analyzed by liquid chromatography-tandem mass spectrometry. The four peptides, pergidin (Perg), 4-valinepergidin (VPerg), dephosphorylated pergidin (dpPerg), and lophyrotomin (LGln), were detected in each of the three stages. Peptide concentrations, in percentage fresh weight, increased from larval up to adult stages, with mean values from 0.044 to 0.125% for Perg, 0.008 to 0.023% for VPerg, and 0.064 to 0.116% for LGln, whereas dpPerg never exceeded 0.001%. The concentrations of this latter peptide averaged 0.002% in the cocoon built by the prepupa, and nearly no peptides were detected in larval feces. Moreover, the concentrations of the three main peptides (Perg, LGln, and VPerg) tended to be correlated with each other in larvae and especially in adults. It is likely that peptide production, purportedly by an endosymbiont, stops at prepupal stage and that concentration of the peptides increases from prepupa to adult due to a decrease of body weight.

Diversity of peptidic and proteinaceous toxins from social Hymenoptera venoms.

Among venomous animals, Hymenoptera have been suggested as a rich source of natural toxins. Due to their broad ecological diversity, venom from Hymenoptera insects (bees, wasps and ants) have evolved differentially thus widening the types and biological functions of their components. To date, insect toxinology analysis have scarcely uncovered the complex composition of bee, wasp and ant venoms which include low molecular weight compounds, highly abundant peptides and proteins, including several allergens. In Hymenoptera, these complex mixtures of toxins represent a potent arsenal of biological weapons that are used for self-defense, to repel intruders and to capture prey. Consequently, Hymenoptera venom components have a broad range of pharmacological targets and have been extensively studied, as promising sources of new drugs and biopesticides. In addition, the identification and molecular characterization of Hymenoptera venom allergens have allowed for the rational design of component-resolved diagnosis of allergy, finally improving the outcome of venom immunotherapy (VIT). Until recently, a limited number of Hymenoptera venoms had been unveiled due to the technical limitations of the approaches used to date. Nevertheless, the application of novel techniques with high dynamic range has significantly increased the number of identified peptidic and proteinaceous toxins. Considering this, the present review summarizes the current knowledge about the most representative Hymenoptera venom peptides and proteins which are under study for a better understanding of the insect-caused envenoming process and the development of new drugs and biopesticides.

Antioxidant and cytotoxic activity of propolis of Plebeia droryana and Apis mellifera (Hymenoptera, Apidae) from the Brazilian Cerrado biome.

Propolis is a complex bioactive mixture produced by bees, known to have different biological activities, especially in countries where there is a rich biodiversity of plant species. The objective of this study was to determine the chemical composition and evaluate the antioxidant and cytotoxic properties of Brazilian propolis from the species Plebeia droryana and Apis mellifera found in Mato Grosso do Sul, Brazil. In the ethanolic extracts of P. droryana propolis (ExEP-P) and A. mellifera (ExEP-A) acids, phenolic compounds, terpenes and tocopherol were identified as major compounds. Both extracts presented antioxidant activity against the 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical, the maximum activities being 500 µg/mL (ExEP-P) and 300 µg/mL (ExEP-A). However, only ExEP-A was able to inhibit lipid peroxidation induced by the oxidizing agent 2,2'-azobis(2-amidinopropane) dihydrochloride (AAPH), which inhibited oxidative hemolysis and reduced the levels of malondialdehyde (MDA) in human erythrocytes for 4 h of incubation. The extracts also reduced the cell viability of the K562 erythroleukemia tumour line, with a predominance of necrotic death. Thus, it is concluded that the propolis produced by P. droryana and A. mellifera contain important compounds capable of minimizing the action of oxidizing substances in the organism and reducing the viability of erythroleukemia cells.

Evolution of Cuticular Hydrocarbons in the Hymenoptera: a Meta-Analysis.

Chemical communication is the oldest form of communication, spreading across all forms of life. In insects, cuticular hydrocarbons (CHC) function as chemical cues for the recognition of mates, species, and nest-mates in social insects. Although much is known about the function of individual hydrocarbons and their biosynthesis, a phylogenetic overview is lacking. Here, we review the CHC profiles of 241 species of Hymenoptera, one of the largest and most important insect orders, which includes the Symphyta (sawflies), the polyphyletic Parasitica (parasitoid wasps), and the Aculeata (wasps, bees, and ants). We investigated whether these taxonomic groups differed in the presence and absence of CHC classes and whether the sociality of a species (solitarily vs. social) had an effect on CHC profile complexity. We found that the main CHC classes (i.e., n-alkanes, alkenes, and methylalkanes) were all present early in the evolutionary history of the Hymenoptera, as evidenced by their presence in ancient Symphyta and primitive Parasitica wasps. Throughout all groups within the Hymenoptera, the more complex a CHC the fewer species that produce it, which may reflect the Occam's razor principle that insects' only biosynthesize the most simple compound that fulfil its needs. Surprisingly, there was no difference in the complexity of CHC profiles between social and solitary species, with some of the most complex CHC profiles belonging to the Parasitica. This profile complexity has been maintained in the ants, but some specialization in biosynthetic pathways has led to a simplification of profiles in the aculeate wasps and bees. The absence of CHC classes in some taxa or species may be due to gene silencing or down-regulation rather than gene loss, as demonstrated by sister species having highly divergent CHC profiles, and cannot be predicted by their phylogenetic history. The presence of highly complex CHC profiles prior to the vast radiation of the social Hymenoptera indicates a 'spring-loaded' system where the diversity of CHC needed for the complex communication systems of social insects were already present for natural selection to act upon, rather than having evolved independently. This diversity may have aided the multiple independent evolution of sociality within the Aculeata.

Toxic peptides occur frequently in pergid and argid sawfly larvae.

Toxic peptides containing D-amino acids are reported from the larvae of sawfly species. The compounds are suspected to constitute environmental contaminants, as they have killed livestock grazing in areas with congregations of such larvae, and related larval extracts are deleterious to ants. Previously, two octapeptides (both called lophyrotomin) and three heptapeptides (pergidin, 4-valinepergidin and dephosphorylated pergidin) were identified from three species in the family Pergidae and one in Argidae. Here, the hypothesis of widespread occurrence of these peptides among sawflies was tested by LC-MS analyses of single larvae from eight pergid and 28 argid species, plus nine outgroup species. At least two of the five peptides were detected in most sawfly species, whereas none in any outgroup taxon. Wherever peptides were detected, they were present in each examined specimen of the respective species. Some species show high peptide concentrations, reaching up to 0.6% fresh weight of 4-valinepergidin (1.75 mg/larva) in the pergid Pterygophorus nr turneri. All analyzed pergids in the subfamily Pterygophorinae contained pergidin and 4-valinepergidin, all argids in Arginae contained pergidin and one of the two lophyrotomins, whereas none of the peptides was detected in any Perginae pergid or Sterictiphorinae argid (except in Schizocerella pilicornis, which contained pergidin). Three of the four sawfly species that were previously known to contain toxins were reanalyzed here, resulting in several, often strong, quantitative and qualitative differences in the chemical profiles. The most probable ecological role of the peptides is defense against natural enemies; the poisoning of livestock is an epiphenomenon.

An insect defensin-derived ß-hairpin peptide with enhanced antibacterial activity.

Insect defensins are a class of small, cysteine-rich antimicrobial peptides primarily active on Gram-positive bacteria. Their roles in maggot therapy for treating chronic wound infection have been reported recently. However, a relatively narrow antibacterial spectrum together with the lack of a cost-effective means of commercial-scale production has limited their application. To further exploit the therapeutic potential of these molecules, we engineered the carboxyl-terminal ß-sheet of navidefensin2-2, an insect defensin from Nasonia vitripennis, based on its structural similarity to naturally occurring microbicidal ß-hairpin peptides. The designed peptide of 14 residues, referred to as NvBH, spans the ß-sheet region of the defensin with two amino acids substituted for assembly of a disulfide-bonded amphipathic ß-hairpin structure. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) combined with circular dichroism (CD) analysis shows that the oxidized NvBH (oNvBH), produced from the synthetic peptide by air oxidization in an alkaline environment, folds into a typical ß-hairpin structure linked by two disulfide bridges (Cys1-Cys4; Cys2-Cys3). However, such a structure appears not to be functionally necessary as synthetic NvBH with a spontaneously oxidized disulfide bridge (Cys2-Cys3) (termed poNvBH) displayed similar antibacterial potency to oNvBH. In comparison with oNvBH, poNvBH exhibited higher serum stability and more resistance on tryptic digestion. These two forms of peptides are capable of killing an array of Gram-positive (including antibiotic-resistant strains of Staphylococcus) and Gram-negative bacterial pathogens at low micromolar concentrations through a membrane disruptive mode of action. Our work indicates that the ß-sheet region of insect defensins is a promising subdomain of proteins in anti-infective drug discovery.

Biomarker genes highlight intraspecific and interspecific variations in the responses of Pinus taeda L. and Pinus radiata D. Don to Sirex noctilio F. acid gland secretions.

Sirex noctilio F., a Eurasian horntail woodwasp recently introduced into North America, oviposits in pines and other conifers and in the process spreads a phytopathogenic fungus that serves as a food source for its larvae. During oviposition the woodwasp also deposits mucus produced in its acid (venom) gland that alters pine defense responses and facilitates infection by the fungus. A 26,496-feature loblolly pine cDNA microarray was used to survey gene expression of pine tissue responding to S. noctilio venom. Six genes were selected for further assessment by quantitative real-time polymerase chain reaction (qRT-PCR), including one that encoded an apparent PR-4 protein and another that encoded a thaumatin-like protein. Expression of both was strongly induced in response to venom, while expression of an apparent actin gene (ACT1) was stable in response to the venom. The pattern of gene response was similar in Pinus taeda L. and Pinus radiata D. Don, but the magnitude of response in P. radiata was significantly stronger for each of the induced genes. The magnitude of the biomarker gene response to venom also varied according to genotype within these two species. The qRT-PCR assay was used to demonstrate that the primary bioactive component in S. noctilio venom is a polypeptide.

Chemometric analysis of Hymenoptera toxins and defensins: A model for predicting the biological activity of novel peptides from venoms and hemolymph.

When searching for prospective novel peptides, it is difficult to determine the biological activity of a peptide based only on its sequence. The "trial and error" approach is generally laborious, expensive and time consuming due to the large number of different experimental setups required to cover a reasonable number of biological assays. To simulate a virtual model for Hymenoptera insects, 166 peptides were selected from the venoms and hemolymphs of wasps, bees and ants and applied to a mathematical model of multivariate analysis, with nine different chemometric components: GRAVY, aliphaticity index, number of disulfide bonds, total residues, net charge, pI value, Boman index, percentage of alpha helix, and flexibility prediction. Principal component analysis (PCA) with non-linear iterative projections by alternating least-squares (NIPALS) algorithm was performed, without including any information about the biological activity of the peptides. This analysis permitted the grouping of peptides in a way that strongly correlated to the biological function of the peptides. Six different groupings were observed, which seemed to correspond to the following groups: chemotactic peptides, mastoparans, tachykinins, kinins, antibiotic peptides, and a group of long peptides with one or two disulfide bonds and with biological activities that are not yet clearly defined. The partial overlap between the mastoparans group and the chemotactic peptides, tachykinins, kinins and antibiotic peptides in the PCA score plot may be used to explain the frequent reports in the literature about the multifunctionality of some of these peptides. The mathematical model used in the present investigation can be used to predict the biological activities of novel peptides in this system, and it may also be easily applied to other biological systems.

Pathological and ultrastructural changes in cultured cells induced by venom from the ectoparasitic wasp Nasonia vitripennis (Walker) (Hymenoptera: Pteromalidae).

The ectoparasitic wasp Nasonia vitripennis produces a proteinaceous venom that induces death in fly hosts by non-paralytic mechanisms. Previous in vitro assays have suggested that the primary cause of cell and tissue death is oncosis, a non-programmed cell death (PCD) pathway characterized by cellular swelling and lysis. However, ultrastructural analyses of BTI-TN-5B1 cells exposed to LC(99) doses of wasp venom revealed cellular changes more consistent with apoptosis and/or non-apoptotic PCD than oncosis or necrosis: By 3h after incubation with venom, susceptible cells displayed indentations in the nuclear membranes, large nucleoli, and extensive vacuolization throughout the cytoplasm. In the vast majority of venom treated cells, annexin V bound to the plasma membrane surface within 15 min after treatment, a characteristic consistent with translocation of phosphatidylserine to the cell surface during the early stages of apoptosis. Likewise, mitochondrial transmembrane potential was depressed in cells within 15 min in venom-treated cells, an event that occurred in the absence of mitochondrial swelling or rupturing of cristae. Active caspase 3 was detected by fluorescent labeling in nearly all venom treated cells 3h after exposure to venom, and in turn, the potent caspase 3 inhibitor Z-VAD-FMK attenuated the morphological changes elicited by wasp venom and afforded protection to BTI-TN-5B1-4 cells.

Antimicrobial peptide-like genes in Nasonia vitripennis: a genomic perspective.

BACKGROUND: Antimicrobial peptides (AMPs) are an essential component of innate immunity which can rapidly respond to diverse microbial pathogens. Insects, as a rich source of AMPs, attract great attention of scientists in both understanding of the basic biology of the immune system and searching molecular templates for anti-infective drug design. Despite a large number of AMPs have been identified from different insect species, little information in terms of these peptides is available from parasitic insects. RESULTS: By using integrated computational approaches to systemically mining the Hymenopteran parasitic wasp Nasonia vitripennis genome, we establish the first AMP repertoire whose members exhibit extensive sequence and structural diversity and can be distinguished into multiple molecular types, including insect and fungal defensin-like peptides (DLPs) with the cysteine-stabilized alpha-helical and beta-sheet (CSalphabeta) fold; Pro- or Gly-rich abaecins and hymenoptaecins; horseshoe crab tachystatin-type AMPs with the inhibitor cystine knot (ICK) fold; and a linear alpha-helical peptide. Inducible expression pattern of seven N. vitripennis AMP genes were verified, and two representative peptides were synthesized and functionally identified to be antibacterial. In comparison with Apis mellifera (Hymenoptera) and several non-Hymenopteran model insects, N. vitripennis has evolved a complex antimicrobial immune system with more genes and larger protein precursors. Three classical strategies that are likely responsible for the complexity increase have been recognized: 1) Gene duplication; 2) Exon duplication; and 3) Exon-shuffling. CONCLUSION: The present study established the N. vitripennis peptidome associated with antimicrobial immunity by using a combined computational and experimental strategy. As the first AMP repertoire of a parasitic wasp, our results offer a basic platform for further studying the immunological and evolutionary significances of these newly discovered AMP-like genes in this class of insects.

Isolation and molecular cloning of venom peptides from Orancistrocerus drewseni (Hymenoptera: Eumenidae).

Three venom peptides (OdVP1, OdVP2 and OdVP3) were isolated from the venom of the solitary wasp Orancistrocerus drewseni (Hymenoptera: Eumenidae). The mature venom peptide sequences were determined via ESI-Q-TOF MS/MS and by searching the O. drewseni venom gland/sac-specific EST library. All of the OdVPs shared the typical characteristics of amidated C-termini proteins and contained a high content of hydrophobic and positively charged amino acids, suggesting that they adopt an amphipathic alpha-helical secondary structure, as is the case for mastoparan from Vespula lewisii. The cDNA sequence of the OdVP1 precursor was obtained by 5'- and 3'-rapid amplification of cDNA ends (RACE), and the OdVP2 and OdVP3 precursor transcripts were identified from the venom gland/sac-specific EST library. While the mature peptide sequences were distinct from one another, the overall transcript structure of the OdVPs showed a high homology to that of mastoparan-B from Vespa basalis in that they contained a signal sequence, a prosequence, a mature peptide and a C-terminal glycine. The OdVPs, particularly OdVP2 and OdVP2L (an analog of OdVP2), exhibited strong antifungal activities, but poor antibacterial activities. OdVP2L, which possessed additional Glu-Pro residues, did not have antimicrobial activity against bacteria or Gram-positive yeast but retained activity against Botrytis cinerea.

Cephalotes clypeatus Fabricius (Hymenoptera: Formicidae): hábitos de nidificação e ocorrência em carcaça animal / Cephalotes clypeatus Fabricius (Hymenoptera: Formicidae): nesting habits and occurrence in animal carcass

A posição ecológica dos himenópteros da família Formicidae em carcaças animais varia de predador, ao alimentar-se de ovos, larvas e pupas de alguns insetos, a necrófago, quando se alimentam de exudatos ou dos tecidos em decomposição. As formigas estão presentes em cadáveres humanos sujeitos a análises forenses e podem inclusive ser úteis na determinação do intervalo post-mortem (IPM). Cephalotes clypeatus Fabricius é exclusivamente arborícola, e ocorre apenas no continente americano. Durante experimento realizado em dezembro de 2003 no campus da Universidade Estadual de Campinas, uma carcaça de camundongo (Mus musculus, linhagem Swiss) de 35,9 g foi exposta em aparato adequado para coleta de exemplares de formigas adultas. A carcaça se decompôs totalmente em quatro dias. Foram coletados 82 exemplares adultos de C. clypeatus, nos dois primeiros dias de exposição. Os indivíduos de C. clypeatus foram observados alimentando-se dos exudatos, tecidos da carcaça e de larvas de dípteros que se encontravam na carcaça. O himenóptero em questão nidificou em galhos ocos de Senna multijuga (Rich.) H.S. Irwin & Barneby (Caesalpinaceae), que se localizava a 1 m do aparato que continha a carcaça. Estudos quanto à biologia desse Cephalotini são necessários, para que se possa ter noção exata do papel da espécie na utilização dos recursos de carcaças animais, e, portanto, no processo de sucessão entomológica. Trata-se do primeiro registro de C. clypeatus em carcaças animais.

The ecological position of the family Formicidae in animal carcasses varies from predator, when feeding on eggs, larvae and pupae of some insects to necrophagous, when the ants feed on exudates or decomposing tissues. Ants are present in human corpses subject to forensic analyses and can also be used in estimation of the post-mortem interval (PMI). Cephalotes clypeatus Fabricius is exclusively arboricolous and occurs only in the American continent. During a field study conducted in the Campus of the Universidade Estadual de Campinas, in December 2003, a laboratory mouse carcass weighing 35,9 g was placed in an iron-mesh cage, which was adequate to collect adult ants. The carcass decomposed in four days. The total of 82 specimens of C. clypeatus was collected, in the first two days of exposure. They were observed feeding on exudates, tissues of the carcass, and on Diptera larvae occurring in the carcass. This species was observed nesting in hollow branches of Senna multijuga (Rich.) H.S. Irwin & Barneby (Caesalpinaceae), which was found one-meter far from the cage. Further investigation on the biology of this Cephalotini must be performed, in order to understand the role of this species in the utilization of animal carcasses, and in the entomological succession process as well. This is the first report of C. clypeatus in animal carcasses.

Parasitoid wasp affects metabolism of cockroach host to favor food preservation for its offspring.

Unlike predators, which immediately consume their prey, parasitoid wasps incapacitate their prey to provide a food supply for their offspring. We have examined the effects of the venom of the parasitoid wasp Ampulex compressa on the metabolism of its cockroach prey. This wasp stings into the brain of the cockroach causing hypokinesia. We first established that larval development, from egg laying to pupation, lasts about 8 days. During this period, the metabolism of the stung cockroach slows down, as measured by a decrease in oxygen consumption. Similar decreases in oxygen consumption occurred after pharmacologically induced paralysis or after removing descending input from the head ganglia by severing the neck connectives. However, neither of these two groups of cockroaches survived more than six days, while 90% of stung cockroaches survived at least this long. In addition, cockroaches with severed neck connectives lost significantly more body mass, mainly due to dehydration. Hence, the sting of A. compressa not only renders the cockroach prey helplessly submissive, but also changes its metabolism to sustain more nutrients for the developing larva. This metabolic manipulation is subtler than the complete removal of descending input from the head ganglia, since it leaves some physiological processes, such as water retention, intact.

Unique toxic peptides isolated from sawfly larvae in three continents.

D-Amino acid containing peptides have been found to be responsible for sawfly larvae poisoning in many parts of the world. These compounds, unique in the animal kingdom, were isolated from three different species of sawfly indigenous to Australia, Denmark and South America. The octapeptide, lophyrotomin, is the major toxin in the Australian and Danish species and is present in small amounts in the South American sawfly. Pergidin, the main toxin in the South American sawfly, is a heptapeptide containing a phosphoseryl residue. This, as far as we are aware, is the first example of such a peptide to be isolated from an animal source. Small amounts of pergidin have been found in the other two species. All available evidence suggests that both peptides are biosynthesised 'de novo' possibly as a protective device, however it cannot be excluded that microorganisms may be responsible. These compounds are stable to enzymatic breakdown because of their configuration and their strong chemical bonding and lipophilic character provide a potential for residues to remain in the host animal and cause significant changes.