Evaluation of cold storage techniques to improve mass rearing of Cleruchoides noackae from Thaumastocoris peregrinus eggs.

The egg parasitoid Cleruchoides noackae Lin & Huber, 2007 (Hymenoptera: Mymaridae) is originated from Australia and the main biological control agent of Thaumastocoris peregrinus Carpenter & Dellapé, 2006 (Hemiptera: Thaumastocoridae) on Eucalyptus L'Hér (Myrtaceae). Companies that grow Eucalyptus are in need of a mass rearing protocol to increase the number of individuals produced and improve the quality of this parasitoid. The aim of this study was to define a protocol for mass rearing C. noackae in T. peregrinus eggs, based in the evaluations of the key biological attributes of this parasitoid in the parental and F1 generations, after the cold storage of the parasitised host eggs. Two methods were tested as C. noackae rearing protocols. In the first, parasitised eggs of T. peregrinus by C. noackae were cold stored for 7 days after being left in a climatic chamber at 24 ± 2°C, 60 ± 10% RH and a photoperiod of 12:12 (light:dark) h (standard environmental conditions) for 3, 6, 9 or 12 days. In the second, T. peregrinus eggs parasitised by C. noackae were maintained in a climatic chamber under standard environmental conditions for 6 days, after which these eggs were cold-stored for 0 (control), 7, 14 or 21 days. Parasitism (%), and the development period (parasitism to adult) and female proportion (%) of C. noackae were evaluated. Based on the results (parental generation: parasitism, around 45%; F1 generation: parasitism, around 55%; development period, around 16 days; female proportion, around 60%), eggs should be stored at 5°C on the sixth day after parasitism by C. noackae and maintained at this temperature for 7 days. The cold storage of T. peregrinus eggs, after parasitism, can be included in the mass rearing protocols of the parasitoid C. noackae.

Side-effects of pesticides on non-target insects in agriculture: a mini-review.

Climate change mediated by anthropogenic activity induces significant alterations on pest abundance and behavior and a potential increase in the use of agrochemicals for crop protection. Pesticides have been a tool in the control of pests, diseases, and weeds of agricultural systems. However, little attention has been given to their toxic effects on beneficial insect communities that contribute to the maintenance and sustainability of agroecosystems. In addition to pesticide-induced direct mortality, their sublethal effects on arthropod physiology and behavior must be considered for a complete analysis of their impact. This review describes the sublethal effects of pesticides on agriculturally beneficial insects and provides new information about the impacts on the behavior and physiology of these insects. The different types of sublethal effects of pesticides used in agriculture on pollinators, predators, parasitoids, and coprophagous insects were detailed.

Side effects of Bacillus thuringiensis on the parasitoid Palmistichus elaeisis (Hymenoptera: Eulophidae).

The endoparasitoid wasp Palmistichus elaeisis Delvare & LaSalle (Hymenoptera: Eulophidae) is used to control defoliating lepidopteran pests. Chemical insecticides are not compatible with natural enemies, but bioinsecticides, such as Bacillus thuringiensis Berliner (Bt), have great potential for use in integrated pest management. However, interactions between Bt and P. elaeisis still need to be investigated. This study aimed to evaluate the effects of Bt on parental and first-generation P. elaeisis parasitizing Bt-susceptible and -resistant Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae). An additional aim was to determine the toxicity of Bt to susceptible third-instar S. frugiperda larvae. Larvae were exposed to lethal concentrations (LC50 and LC90) of Bt and then allowed to be parasitized by P. elaeisis. Parasitoid longevity, immature production, reproductive performance, and behavioral responses were evaluated. Bt repelled P. elaeisis and reduced immature production. Parental and first filial generation parasitoids of both sexes emerged from Bt-treated larvae showed lower survivorship than controls. Parasitoids had poorer reproductive performance in Bt-susceptible and -resistant pupae than in untreated pupae. Palmistichus elaeisis emerged from Bt-susceptible and -resistant S. frugiperda showed altered host-searching behavior and reproductive parameters, which indicates low compatibility between the bioinsecticide agent and the parasitoid wasp.

Phytoremediation by Eremanthus crotonoides and Inga striata decay atrazine and clomazone residues in the soil.

The atrazine and clomazone molecules have potential to contaminate environments, especially water resources. Phytoremediation decontamination can prevent or reduce the quantity of these products reaching watercourses. The objective was to evaluate the remedial potential and the physiological sensitivity of Eremanthus crotonoides (DC.) Sch. Bip and Inga striata Benth to atrazine and clomazone in soils contaminated with 0.0, 0.5, 1.0 and 2.0 times the recommended commercial dose of these herbicides. The photosynthesis, CO2 consumed stomatal conductance and transpiration of E. crotonoides and I. striata, in soils contaminated with atrazine and clomazone, were evaluated. The herbicide residues were detected by ultra-high performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) 30 days after the last herbicide application. The photosynthesis rate and CO2 consumed by E. crotonoides and I. striata were lower in soils contaminated by atrazine and clomazone. Inga striata had lower stomatal conductance and transpiration in soil contaminated with clomazone. Eremanyhus crotonoides and I. striata reduced the residues of these herbicides. The atrazine and clomazone reduced the physiological variables of E. crotonoides and I. striata. These plants can be used to recover areas with residues of these herbicides, acting as filters that will decrease the amount of herbicides that would reach the watercourses.

Toxicity and cytotoxicity of the insecticide imidacloprid in the midgut of the predatory bug, Podisus nigrispinus.

The selectivity of insecticides on natural enemies in pest control are an important strategy for Integrated Pest Management. However, insecticides can have side effects on non-target organisms such as natural enemies. This study evaluated the histological and cytological changes mediated by the sublethal concentration of the imidacloprid insecticide on the midgut of non-target predator Podisus nigrispinus (Heteroptera: Pentatomidae), used in the biological control of pests. Imidacloprid was toxic for P. nigrispinus with LC50 = 3.75 mg L-1 and survival of 51.8%. This sublethal concentration of imidacloprid causes histological alterations in the midgut epithelium and cytotoxic features were irregular border epithelium, cytoplasmic vacuolation, and apocrine secretions in the first 6 h after exposure with the insecticide. Apoptosis in the digestive cells occurs after 12 h of exposure in the midgut. These results suggest that imidacloprid may affect the digestive physiology of P. nigrispinus and compromise the effective predation of this insect a biological control agent. The associated use of this insecticide with the predator in pest control should be carefully evaluated.

Chlorantraniliprole-mediated effects on survival, walking abilities, and respiration in the coffee berry borer, Hypothenemus hampei.

Hypothenemus hampei Ferrari (Coleoptera: Curculionidae) is the main pest of coffee crops, and effective methods for pest management are needed urgently. Bioassays were conducted to assess the effects of the insecticide chlorantraniliprole on H. hampei adults. Toxicity, survivorship, larval production, respiration rate, and behavioral responses to six concentrations of chlorantraniliprole were evaluated. Chlorantraniliprole was toxic to H. hampei (LD50 = 0.49 mg mL-1 and LD90 = 1.21 mg mL-1). Survivorship was 98% in adults not exposed to chlorantraniliprole, decreasing to 52% in insects exposed to LD50 and 2% in insects treated with LD90. H. hampei showed reduced mobility on insecticide-treated surfaces. The insecticide promoted a decrease in the respiration rate of H. hampei for up to 3 h after exposure, altering behavioral responses and locomotor activity. Chlorantraniliprole was shown to have lethal and sublethal effects on H. hampei and, thus, can be used rotationally in integrated pest management programs to control of this pest in coffee crops and retard of insect resistance.

Effects of the herbicide trifluralin in the initial development of Piptadenia gonoacantha (Fabales: Fabaceae).

Trifluralin, a pre-emergent herbicide, is widely used in Brazil in the weed grass management in restoration areas. The objective was to evaluate the tolerance of Piptadenia gonoacantha to trifluralin. The treatments had three trifluralin doses (445, 890, and 1335 g a.i. ha-1), applied in pre-sowing, as well as the control, without herbicide. Visual intoxication, seed germination, survival rate, emergence speed index (EMI), mean germination period, seedling height, and diameter, micromorphometric parameters of plant roots collected at 60 d after sowing, root length (RL) and volume, leaf area (LA), leaf numbers, root and shoot dry matter, and fluorescence of chlorophyll a at 30, 45, and 60 d after sowing were analyzed. Visual intoxication values above 50% were observed only with 1335 g a.i. ha-1. The herbicide did not affect seed germination, EMI, average germination period, seedling height, and diameter, root micromorphometric parameters, length, dry matter or root volume, and chlorophyll a fluorescence. The dose 1335 g a.i. ha-1 caused a reduction of 41.5% in survival, 50.3% in the LA, 36.7% in the number of leaves (LN), and 59.8% in the aerial dry mass of seedlings. The trifluralin presents potential for restoration programs of degraded areas with this forest species.

Post-embryonic changes in the hindgut of honeybee Apis mellifera workers: Morphology, cuticle deposition, apoptosis, and cell proliferation.

In insects, the hindgut is a homeostatic region of the digestive tract, divided into pylorus, ileum, and rectum, that reabsorbs water, ions, and small molecules produced during hemolymph filtration. The hindgut anatomy in bee larvae is different from that of adult workers. This study reports the morphological changes and cellular events that occur in the hindgut during the metamorphosis of the honeybee Apis mellifera. We describe the occurrence of autophagosomes and the ultrastructure of the epithelial cells and cuticle, suggesting that cuticular degradation begins in prepupae, with the cuticle being reabsorbed and recycled by autophagosomes in white- and pink-eyed pupae, followed by the deposition of new cuticle in light-brown-eyed pupae. In L5S larvae and prepupae, the hindgut undergoes cell proliferation in the anterior and posterior ends. In the pupae, the pylorus, ileum, and rectum regions are differentiated, and cell proliferation ceases in dark-brown-eyed pupae. Apoptosis occurs in the hindgut from the L5S larval to the pink-eyed pupal stage. In light-brown- and dark-brown-eyed pupae, the ileum epithelium changes from pseudostratified to simple only after the production of the basal lamina, whereas the rectal epithelium is always flattened. In black-eyed pupae, ileum epithelial cells have large vacuoles and subcuticular spaces, while in adult forager workers these cells have long invaginations in the cell apex and many mitochondria, indicating a role in the transport of compounds. Our findings show that hindgut morphogenesis is a dynamic process, with tissue remodeling and cellular events taking place for the formation of different regions of the organ, the reconstruction of a new cuticle, and the remodeling of visceral muscles.

Terpenoid constituents of cinnamon and clove essential oils cause toxic effects and behavior repellency response on granary weevil, Sitophilus granarius.

This study evaluated toxic effects, repellency and respiration rate caused by terpenoid constituents of cinnamon and clove essential oils and against Sitophilus granarius L. (Coleoptera: Curculionidae). The lethal concentrations (LC50 and LC90), repellent effect, and behavior repellency response on adults of S. granarius after exposure to six concentrations of each essential oil and terpenoids were evaluated. The chemical composition of the cinnamon oil was also determined and primary compounds were eugenol (10.5%), trans-3-caren-2-ol (10.2%), benzyl benzoate (9.99%), caryophyllene (9.34%), eugenyl acetate (7.71%), α-phellandrene (7.41%), and α-pinene (7.14%). In clove essential oil, the primary compounds were eugenol (27.1%), caryophyllene (24.5%), caryophyllene oxide (18.3%), 2-propenoic acid (12.2%), α-humulene (10.8%), γ-cadinene (5.01%), and humulene oxide (4.84%). Cinnamon and clove essential oil was toxic to S. granarius. In toxic terpenoids compounds, eugenol has stronger contact toxicity in S. granarius than caryophyllene oxide, followed by α-pinene, α-humulene, and α-phellandrene. Insects reduced their respiratory rates after being exposed to essential oil terpenoids and avoided or reduced their mobility on terpenoid-treated surfaces. Cinnamon and clove essential oil, and their terpenoid constituents were toxic and repellent to adult S. granarius and, therefore, have the potential to prevent or retard the development of insecticide resistance.

Squamocin induce histological and ultrastructural changes in the midgut cells of Anticarsia gemmatalis (Lepidoptera: Noctuidae).

Annonaceous acetogenins (Annona squamosa Linnaeus) comprises of a series of natural products which are extracted from Annonaceae species, squamocin proved to be highly efficient among those agents. Squamocin is mostly referred as a lethal agent for midgut cells of different insects, with toxic effects when tested against larva of some insects. In present study, LC50 and LC90 of squamocin for A. gemmatalis Hübner (Lepidoptera: Noctuidae) were calculated using probit analysis. Morphological changes in midgut cells were analyzed under light, fluorescence and transmission electron microscopes when larvae were treated with LC50 and LC90 of squamocin for 24, 48 and 72 h. Results revealed that the maximum damage to midgut cells was found under LC90 where it showed digestive cells with enlarged basal labyrinth, highly vacuolated cytoplasm, damaged apical surface, cell protrusions to the gut lumen, autophagy and cell death. The midgut goblet cells showed a strong disorganization of their microvilli. Likewise, in insects treated with squamocin, mitochondria were not marked with Mitotracker fluorescent probe, suggesting some molecular damage in these organelles, which was reinforced by decrease in the respiration rate in these insects. These results demonstrate that squamocin has potential to induce enough morphological changes in midgut through epithelial cell damage in A. gemmatalis.

Exposure to the insecticide endosulfan induces liver morphology alterations and oxidative stress in fruit-eating bats (Artibeus lituratus).

Exposure to pesticides may increase the generation of reactive oxygen species (ROS), leading to oxidation of cell membrane lipids and proteins. Although fruit bats are potentially exposed to pesticides during their entire lifespan, the impacts of this exposure are still poorly investigated. We examined the effects of low, commercially recommended concentrations (0, 1.05 and 2.1 g/l) of an organochlorine insecticide endosulfan (EDS) formulation on oxidative responses in the liver and kidneys of Neotropical fruit bats (Artibeus lituratus), as well as possible liver morphological alterations following a 35-day oral exposure. Superoxide dismutase activity was significantly decreased upon exposure to 1.05 g/l of EDS in the liver and kidneys, catalase was decreased in the liver of 2.1 g/l EDS-exposed bats, while glutathione S-transferase was increased in the liver of 2.1 g/l EDS-exposed bats. Protein carbonyls increased following the exposure to the highest EDS dose tested. Endosulfan-induced morphological alterations in the liver included cell degeneration and cell death, with apparent cytoplasm lipid accumulation (steatosis) and pyknotic nuclei, karyolysis and deposit of collagen fibres. Our findings suggest that exposure to low concentrations of EDS induced a certain extent of oxidative damage in fruit bats, which may have led to liver morphological alterations.

Feeding habits of marmosets: A case study of bark anatomy and chemical composition of Anadenanthera peregrina gum.

Primates of the genus Callithrix often obtain exudates from plants of the family Fabaceae. This study characterizes the chemical composition of exudates, and the anatomy and hystochemistry of the secretory ducts in the bark of Anadenanthera peregrina (L.) Speg. var. peregrina (Fabaceae). Exudates from this tree species represent an important component of the diet of hybrid marmosets, Callithrix spp. (Primates: Cebidae). A. peregrina was selected as the focal study tree because it is the only gum tree species exploited by Callithrix groups present within five urban forest fragments in the municipality of Viçosa, Minas Gerais State, Brazil. Gum samples were obtained directly from gouges made by the marmosets, while bark samples were obtained from A. peregrina plants, whether or not they were damaged by the marmosets. Constitutive secretory ducts were present in the bark of ungouged A. peregrina, whereas, marmoset damage caused induced secretory duct formation and an increase in the size of these ducts. The gum produced in the gouges made by the marmosets and in ungouged plants reacted positively to tests for polysaccharides, pectin, mucilage, and proteins. The gum from the gouges exhibited high water (41.0%), carbohydrate (38.2%), protein (19.0%), and mineral (Ca 0.4% and K 0.3%) content. We argue that the relatively high calcium content of A. peregrina gum plays an important nutritional role in, balancing a diet that is otherwise rich in phosphorous and poor in calcium.

Nematophagous fungi combinations reduce free-living stages of sheep gastrointestinal nematodes in the field.

Gastrointestinal nematodes (GIN) can reduce or limit sheep production. Currently there is a clear deficiency in the action of drugs for the control of these parasites. Nematophagous fungi are natural enemies of GIN. Fungal combinations have potential for reducing GIN populations. The aim of this study was to evaluate the efficiency combinations of nematophagous fungi in sodium alginate matrix pellets for the biological control agents of gastrointestinal sheep nematode parasites in the field. The nematophagous fungi (0.2mg of fungus per kg of body weight), Arthrobotrys conoides, A. robusta, Duddingtonia flagrans, and Monacrosporium thaumasium were used. The treated groups were administered mycelium combinations in the following combinations: group 1 (D. flagrans+A. robusta); group 2 (M. thaumasium+A. conoides). The control group did not receive any fungal pellets. We used three groups with eight Santa Inês sheep each. Each animal was treated with approximately 1g of pellet per 10kg of live weight. During the experimental period, we evaluated: number of eggs per gram of feces (EPG), infective larvae (L3) per kg of dry matter, larvae recovered from coprocultures, packed cell volume, total plasma protein concentration of sheep, and environmental conditions. Group 2 EPG (M. thaumasium+A. conoides) differed from the control group in September and October. The number of L3/kg of dry matter recovered from animals of groups 1 and 2 at distances of 0-20 and 20-40cm from the fecal pats was lower than the control group. The packed cell volume and total plasma proteins of treated animals were similar to those of the control group. The combination of treatment groups (D. flagrans+A. robusta and M. thaumasium+A. conoides) reduced the number of L3/kg of pasture. Therefore, treatment of nematophagous fungal combinations have the potential to manage free-living stages of GIN in sheep.

Oat, Wheat, and Sorghum Genotype Reactions to Meloidogyne incognita and Meloidogyne javanica.

Meloidogyne spp. are the most economically important species of plant-pathogenic nematodes. Plant resistance and crop rotation are the main nematode management methods. Thus, the objective was to evaluate the resistance of seven wheat genotypes, five oat genotypes, ten sorghum hybrids, and three sorghum-sudangrass genotypes to Meloidogyne incognita and Meloidogyne javanica. The crops were sowed in pots with an autoclaved substrate. A single plant/pot was left after thinning. The soil was infested with 5,000 eggs of the studied nematodes. Tomato (cv. Rutgers) plants were used as the standard for nematode susceptibility. The evaluations were conducted 60 d after inoculation. Gall and egg-mass indexes were obtained according to a 0-5 scale. Plants with a reproduction factor higher than 1.0 were classified as susceptible (S) and lower than 1.0 as resistant (R). Wheat and oat genotypes did not allow M. incognita and M. javanica reproduction, proving resistance to these organisms. Sorghum genotypes had different reactions to M. incognita and M. javanica. The tomato (cv. Rutgers) plants demonstrated the viability of the nematode inoculum for the three crops. The wheat and oat genotypes and the sorghum hybrids 'BRS-610', 'BRS-800', and '307.343' can be used in crop rotation systems for M. incognita and M. javanica management.

Juvenile hormone downregulates vitellogenin production in Ectatomma tuberculatum (Hymenoptera: Formicidae) sterile workers.

In the ant Ectatomma tuberculatum (Olivier 1792), workers have active ovaries and lay trophic eggs that are eaten by the queen and larvae. Vitellogenins are the main proteins found in the eggs of insects and are the source of nutrients for the embryo in the fertilized eggs and for adults in the trophic eggs. In social insects, vitellogenin titres vary between castes and affect reproductive social status, nursing, foraging, longevity, somatic maintenance, and immunity. In most insects, vitellogenin synthesis is mainly regulated by juvenile hormone. However, in non-reproductive worker ants, this relationship is poorly characterized. This study determined the effects of juvenile hormone on vitellogenin synthesis in non-reproductive E. tuberculatum workers. Juvenile hormone was topically applied onto workers, and the effect on vitellogenin synthesis in the fat body and vitellogenin titres in the haemolymph were analysed by ELISA and qPCR. Juvenile hormone downregulated protein synthesis and reduced vitellogenin titres in the haemolymph, suggesting that in workers of E. tuberculatum, juvenile hormone loses its gonadotrophic function.

Antennal Sensilla and Sexual Dimorphism of the Parasitoid Trichospilus pupivorus (Hymenoptera: Eulophidae).

Sensory structures (sensilla) of insects are present in high number in the antennae are important for perceiving environmental cues. Thus, sensilla play an important role in the localization, discrimination, and acceptance of a possible host by parasitoid insects. The sensilla, classified according to their shape, size, and distribution, may also show sexual dimorphism in insects. The types and distribution of antennal sensilla in female and male parasitoid Trichospilus pupivorus (Ferriere, 1930) (Hymenoptera: Eulophidae) were evaluated. T. pupivorus females had longer antennae than males. Both males and females have ten sensilla types with higher abundance of placoid sensilla in females, which suggest a possible functional dimorphism.

Interactions Between the Bud Rot Disease of Oil Palm and Rhynchophorus palmarum (Coleoptera: Curculionidae).

Rhynchophorus palmarum (L.) causes great losses to the oil palm plantations, and therefore, the spatial and temporal distribution of this insect should be studied, to manage its populations. Insect sampling was done for 2 yr in an oil palm plantation from Colombia. In total, 60 pheromone traps were used in healthy palm trees and infected ones with the Bud Rot disease. On the other hand, developmental stages of this insect were quantified on healthy and diseased palms for two consecutive years. Number of adult R. palmarum per sampling was higher in the plantation with diseased palm trees, 3.85 and 74.7 insects per trap, than in those with healthy ones, 1.91 and 9.48 insects per trap, in the first and second years, respectively. After the integration of pheromone traps, there was a significant increase in the infestation level at all stages of development of the insect. For the first time, the presence of R. palmarum attracted to diseased palms is reported. The association between R. palmarum and the Bud Rot disease is a cause of death and great loss to the oil palm plantations.

Persistence of fipronil residues in Eucalyptus seedlings and its concentration in the insecticide solution after treatment in the nursery.

Eucalyptus seedlings are normally protected from underground termites (Isoptera: Termitidae) by immersing them in insecticide solutions. Fipronil (phenylpyrazole) is the most frequently used product to protect seedlings in the field for up to 6 months after application. This is performed just prior to planting. However, the persistence of this product in seedlings that are treated and subjected to irrigation several days prior to planting has not yet been evaluated. This study aims to quantify the fipronil concentration in the substratum and roots of the seedlings treated and subjected to irrigation for up to 56 days prior to planting and to quantify this insecticide concentration in the solutions, without continuous stirring, for 120 min. The quantitative determination of fipronil in the seedlings and in the insecticide solution was done by high-performance liquid chromatography (HPLC) with an ultraviolet (UV) detector. It was found that irrigation up to 56 days, performed in the nurseries, did not decrease the fipronil concentration in the seedlings. The absence of stirring reduced the fipronil concentration in the insecticide solution, necessitating a homogenization system to maintain the recommended concentration of this product, to effectively treat the eucalyptus seedlings. The seedling treatment with fipronil can be conducted strictly in the nursery, reducing cost and environmental risks.

Ultrastructure of the excretory organs of Bombus morio (Hymenoptera: Bombini): bee without rectal pads.

Bumblebees need to keep bodily homeostasis and for that have an efficient system of excretion formed by the Malpighian tubules, ileum, and rectum. We analyzed the excretory organs of Bombus morio, a bee without rectal pads. In addition, we analyzed the rectal epithelium of Melipona quadrifasciata anthidioides which has rectal pads. The Malpighian tubules exhibited two cell types and the ileum four types. However, comparative analysis of the rectum showed that only cells of the anterior region of the rectal epithelium of B. morio are structurally distinct. We suggest that cells of the Malpighian tubules of B. morio have an excretory feature and that cells of ileum have different functions, such as ion absorption and water, organic compound, and protein secretion. In addition, only the anterior region of the rectum of B. morio showed characteristic absorption. We suggest that Malpighian tubules participate in the excretion of solutes and that the ileum and rectal epithelium are responsible for homeostasis of water and solutes, compensating for the absence of rectal papillae. These results contribute to our understanding of the morphophysiology of the excretory organs of bees without rectal pads.

Endocrine and regenerative cells in the midgut of Chagas' disease vector Triatoma vitticeps during different starvation periods.

The midgut of an insect has three main cell types, viz., digestive, endocrine and regenerative cells, and a relationship between cell morphology and blood meal has been suggested. This study evaluated the occurrence of the regenerative and endocrine cells in different regions of the midgut of female and male Triatoma vitticeps during different periods after a blood meal. Adults of both sexes were dissected at 4, 72, 120, 168, 288, 360 and 600 hours after feeding and the midgut was pulled apart. The midgut was divided into the anterior, middle and posterior regions and the fragments were analyzed under a light microscope. The results showed the presence of endocrine cells of the open type and positive for FMRFamide as well as regenerative cell nests in the midgut of T. vitticeps. An association was observed between the starvation period and frequency of FMRFamide positive and regenerative cells in the midgut of T. vitticeps. Differences in the pattern of distribution of these cells between the males and females as well as in the different regions of the midgut were also discussed.