Melatonin associated with a high-fat diet during pregnancy and lactation prevents liver changes in the offspring.

In the present study, we set out to determine whether melatonin combined with a high-fat diet during pregnancy and lactation can prevent liver disorders in offspring. Forty rats were divided into four groups: DC - pregnant rats submitted to the standard diet; DC + Mel - pregnant rats submitted to the standard diet combined with melatonin; HFD - pregnant rats submitted to a high-fat diet; HFD + Mel - pregnant rats submitted to a high-fat diet combined with melatonin. Morphophysiological and biochemical parameters were analyzed. Melatonin (5 mg/kg) was administered intraperitoneally. The HFD group offspring showed an increase in AST, ALT, alkaline phosphatase, cholesterol, triglycerides, LDL and glucose levels, and a reduction in HDL and lipase levels. In the liver obseved steatosis, hepatocellular ballooning, increased lobular parenchyma and reduced non-lobular parenchyma, beside reduced liver glycogen and fibrosis. These changes were not observed in the HFD + Mel group. In conclusion, melatonin combined with a high-fat diet preserves the liver architecture and function in the offspring.

Effect of swimming exercise, insulin-associated or not, on inflammatory cytokines, apoptosis, and collagen in diabetic rat placentas.

Physical exercise is an important therapeutic agent for women with diabetes during gestation. However, its histophysiological consequences for the placenta remain unclear. In this study, we evaluated the expression of VEGF-A, IL1ß, TNFα, and type I collagen in the placentas of diabetic rats subjected to a swimming program. Thirty rats were divided into the following groups: CG, pregnant nondiabetic rats; CEG, nondiabetic pregnant rats subjected to swimming; DG, pregnant diabetic rats; DEG, pregnant diabetic rats subjected to swimming; DIG, pregnant diabetic rats treated with insulin; DIEG, pregnant diabetic rats treated with insulin and subjected to swimming. Diabetes was induced using streptozotocin [50 mg/kg intraperitoneally (i.p.)], and insulin was administered at a dose of 5 U/day i.p. (2 U at 10 am and 3 U at 7 pm) in the DIG group; in the DIEG group, insulin was administered at a dose of only 2 U/day at 7 pm. The rats were sacrificed on the 20th day of gestation. There was an increase in the expression of IL-1ß, TNF-α, VEGF-A, and type I collagen and a higher apoptotic index in the placentas of the DG and DEG groups, but there was a reduction in glycemia in the latter group. In the DIG and DIEG groups, the levels remained similar to those of the control; however, in these groups the reduction was more significant for all analyzed parameters. Therefore, in rats induced to diabetes during pregnancy, swimming, although reducing glycemic levels, did not prevent immunohistochemical changes in the placenta, suggesting the need for a multidisciplinary protocol associated with traditional pharmacological treatment.

Side effects of a mixture of essential oils on Psyttalia concolor.

Integrated Pest Management programs do not always prioritize natural enemies when selecting control methods; too often these important pest reducing agents are negatively affected by the action of other methods in agroecosystems. The aim of this research was to evaluate side effects of a bioinsecticide, developed from the mixture of cedar (Cedrus atlantica), eucalyptus (Corymbia citriodora) and lemon grass (Cymbopogon citratus) essential oils (EOs), in a ratio of 1:1:1, on Psyttalia concolor (Szèpligeti) (Hymenoptera: Braconidae) which is a parasitoid of some pests of the Tephritidae family, as Ceratitis capitata. The LD50 of the EOs mixture for C. capitata females was 3.09 µl/g, whereas the LD50 for P. concolor females was 20.45 µl/g which suggests the natural enemy is more tolerant to the EOs mixture. P. concolor parasitized L3 larvae of C. capitata through a voile treated with the mixture of EOs at 1.8% without causing any deleterious effects neither on the percentage of attacked hosts nor on the emergence rate, whereas at the highest concentration tested, 4.8%, decreased both parameters during the 2 first days after treatment. Semi-field assays showed that mixture of EOs was harmless to P. concolor when released 4 h after treatment whereas killed 55 ± 3.9% males and 37.5 ± 1.6% females of C. capitata. The mixture of EOs used jointly with lambda-cyhalothrin or kaolin, both compounds applied against C. capitata in conventional and organic farming, respectively, did not increase the toxicity and/or persistence against the pest.

Maternal Na+ intake induces renal function injury in rats prevented by a short-term angiotensin converting enzyme inhibitor.

The Na+ -ATPase, a secondary pump in the proximal tubule, is only weakly responsive to angiotensin II in adult offspring exposed perinatally to high Na+ intake. We have investigated whether the offspring from mothers given 0.3 mol/L NaCl show an ineffective angiotensin II action to increase in blood pressure. It was hypothesized that functional alterations at adult life are associated with the number of angiotensin II-positive cells in the developing kidney, with increased oxidative stress in maternal/foetal organs, or with morphometrical changes in placentas. Wistar female rats were maintained on 0.3 mol/L NaCl in their drinking water from 20 days before conception until weaning. After weaning, some of the male offspring were treated with enalapril for 21 days. Glomerular filtration rate was recorded up to 210 days of age, when mean arterial pressure was measured after infusion of angiotensin II. To investigate the placenta and foetal kidneys, mothers on tap water or NaCl were also treated with alpha-tocopherol, pregnancy being interrupted on the 20th day. There were no changes in the number of cells positive for angiotensin II in the foetal kidney and unchanged lipid peroxidation in the placenta of offspring exposed to NaCl, but the intermediate trophoblast area in the junctional zone was increased, possibly reducing maternal-foetal exchange. Glomerular filtration rate was reduced and there was an attenuated effect of angiotensin II on elevation of blood pressure, which could be mediated by an elevated angiotensin II during early life, once these disturbances had been prevented by early and short-term treatment with enalapril.

Effect of the combination of metformin hydrochloride and melatonin on oxidative stress before and during pregnancy, and biochemical and histopathological analysis of the livers of rats after treatment for polycystic ovary syndrome.

The aim of the present study was to analyze the effect of a combination of metformin hydrochloride and melatonin on oxidative stress together with a biochemical and histopathological analysis of the livers of Wistar rats induced with PCOS. The results indicated that a combination of the drugs was more effective in the reduction of plasmatic levels of liver enzyme alanine aminotransferase, nitric oxide and total glutathione, and decreased the inflammatory response and histopathological damage, producing results that were significantly similar to animals from the control group. A mixture of the drugs produced more effective results against liver toxicity caused by PCOS, encouraging the normalization of biochemical parameters. During pregnancy, there was reduced oxidative stress compared to monotherapeutic use of these drugs. Interestingly, the combination of the drugs caused a physiological reaction similar to responses identified in healthy rats without induction of the PCOS control group. However, the clinical and physiological effectiveness of the combination should be further explored, especially with respect to the possible side effects on offspring.

Cotesia flavipes (CAM) (Hymenoptera: Braconidae) supresses immune responses in Diatraea flavipennella (BOX) (Lepidoptera: Crambidae).

The present research aimed to elucidate which aspects of immune responses in Diatraea flavipennella are suppressed by the parasitoid Cotesia flavipes, thus, ensuring parasitism success. We investigated the presence of apoptosis in fat body cells through the TUNEL technique. According to the results, reduced levels of nitric oxide and phenoloxidase activity were observed in larvae parasitized for three days, and reduced total number of hemocytes, after three and seven days. An increase in plasmatocytes and decrease in spherulocytes numbers were observed in the differential count on the third day of parasitism. The number of melanized microspheres in parasitized larvae was low and indicated less intense melanization. The ultrastructural analysis confirmed the immunosuppressive effect of C. flavipes on the encapsulation response of D. flavipennella because only the formation of hemocytes capsules, adhered to the microspheres' surface, was evidenced in non-parasitized caterpillars. The effect of parasitism was also recorded on the third day with the presence of hemocytes and apoptosis in fat body cells, including aspects of degeneration in the latter. We concluded that C. flavipes suppresses cellular and humoral immunological responses in D. flavipennella and drastically affects the host's fat tissue.

Sublethal effect of concentrations of Metarhizium anisopliae (Metsch.) Sorok on the larval stage and immunologic characteristics of Diatraea flavipennella (BOX) (Lepidoptera: Crambidae).

This work aimed to evaluate the effects of Metarhizium anisopliae on Diatraea flavipennella and investigate their immune response. Was observed the effect of M. anisopliae against larvae of D. flavipennella sprayed at concentrations of 103, 104, 105 conidia / mL, in which showed differences relative the larval period, extending up to 72.0 days in treatment and 25.0 days in the control. The results for hemocytes revealed that the most frequent cells when sprayed at the concentrations of 103, 105, 107 conidia / mL were the prohemocytes, spherulocytes, plasmatocytes and granulocytes in relation to adipohemocytes and oenocytoids. The level of nitric oxide was different between the control and the concentration 107 spores / mL (24), while the activity of phenoloxidase was similar among treatments in 24 and higher concentration 107 spores / mL (60h). In biochemical profile of hemocytes was a change in carbohydrates, lipids and proteins in response to the fungus. The results indicate that the fungus M. anisopliae can be used in the Integrated Management of D. flavipennella by presenting pathogenicity and interfere with their development even when exposed to small concentrations.

Microscopic analysis of Spodoptera frugiperda (Lepidoptera: Noctuidae) embryonic development before and after treatment with azadirachtin, lufenuron, and deltamethrin.

The botanical insecticides, growth regulators, and pyrethroids have an effect on the biology of Spodoptera frugiperda (Smith). However, no emphasis has been given to the effect of these insecticides on embryonic development of insects, in histological level. Thus, this research aimed to examine by light and scanning electron microscopy S. frugiperda eggs and to describe the embryonic development, before and after immersion treatment, using commercial concentrations and lower concentrations than commercial ones, of the compounds lufenuron (Match), azadirachtin (AzaMax), and deltamethrin (Decis-positive control). For light microscopy semithin sections of eggs were used, and for scanning electron microscopy, images of the surface of eggs, treated and untreated with insecticides. The morphological characteristics of S. frugiperda eggs, in general, were similar to those described in the literature for most of the insects in the order Lepidoptera. Spherical eggs slightly flattened at the poles, with chorion, yolk, vitelline membrane, and embryo formation. In both microscopic analysis, we observed that insecticides acted immediately and independent of concentration, resulting absence, or incomplete embryo, presented yolk granules widely dispersed, without vitellophage formation, chorion disintegration, disorganized blastoderm, presenting vacuoles, yolk region with amorphous cells, and formation of completely uncharacterized appendages. Thus, we conclude that the compounds lufenuron and azadirachtin interfere on S. frugiperda embryonic development.

Bacillus thuringiensis affects reproductive capacity of adult rat offspring.

We investigated the effects of B. thuringiensis-based biological insecticides, XenTari and Dipel, and deltamethrin on the reproductive development of pups of pregnant rats. Twenty 90-day-old pregnant rats were divided randomly onto four equal groups: control group (GC) administered only water; XenTari group (GX) administered 1 mg XenTari (containing Cry1Ac toxin of B. thuringiensis)/100 g body weight; Dipel group (GDi) administered 1 mg Dipel (containing Cry1Aa, Cry1Ab and Cry1Ac toxins of B. thuringiensis)/100 g body weight; and a deltamethrin group (GDe) administered 2 mg deltamethrin (0.08 ml Keshet 25EC)/kg body weight as a positive control. Insecticides were administered by gavage at doses of 1 mg/100 g/day (GX and GDi), and 2 mg/kg/day (GDe) during pregnancy and lactation. Treatment with both biologic and synthetic insecticides reduced the weight gain of the mothers. The biological insecticides reduced the number, weight and length, and increased malformation and mortality of the offspring. In female offspring for all three groups administered insecticides, opening of the vagina was delayed, metestrus was increased and estrogen and progesterone levels were reduced compared to proestrus, estrus and metestrus of the cycle. The ovaries of female offspring of all three groups administered insecticides contained numerous tertiary and atretic follicles, few corpora lutea, primary and secondary follicles, and reduced estrogen receptors compared to controls. In male offspring, all three groups exposed to insecticides exhibited reduced testosterone levels. Histopathological changes in the testes including vacuolation and desquamation of the seminiferous epithelium were observed only in the GX and GDi groups. The number of androgen receptors was reduced significantly in the testes and testicular morphometry revealed reduced tubule diameter, height of the seminiferous epithelium and total tubule length compared to the control. The biological insecticides, XenTari and Dipel, administered in sublethal doses to pregnant rats, caused reproductive changes in the offspring similar to those of the insecticide, deltamethrin.

Melatonin Administration Prevents Placental and Fetal Changes Induced by Gestational Diabetes.

Gestational diabetes mellitus (GDM) promotes changes in the placenta and fetuses, due to oxidative stress. Antioxidants can reduce oxidative stress in the placenta. We tested the hypothesis that melatonin (Mel) can prevent these effects in the placenta and fetuses, analyzing their histology, histochemistry, morphometry, and immunohistochemistry. Thirty albino rats were used, divided into groups: CG-pregnant non-diabetic rats; GD-pregnant diabetic rats; GD + Mel-pregnant diabetic rats treated with melatonin. Diabetes was induced by streptozotocin at a dosage of 50 mg/kg i.p. Melatonin was administered in daily injections of 0.8 mg/kg i.p. Melatonin prevented the placental weight and fetal weight and length from increasing, in addition to histomoformetric, histochemical, and immunohistochemical changes in the placentas, compared to the placentas of diabetic females (GD). Thus, we conclude that melatonin has a great potential to prevent placental changes due to GDM.

Follicle-stimulating hormone mediates the consumption of serum-derived glycogen by bovine cumulus-oocyte complexes during in vitro maturation.

BACKGROUND AND AIM: Oocyte in vitro maturation (IVM) is an appealing approach for several assisted reproductive technologies and dissecting oocyte maturation. Nonetheless, IVM leads to lower developmental competence and usually relies on undefined, serum-containing media. Therefore, biochemical profiling aimed to explore fluctuations in IVM media content during the acquisition of oocyte developmental competence. MATERIALS AND METHODS: Bovine cumulus-oocyte complexes (COCs) underwent IVM in TCM199 medium with Earle's salts, supplemented with 2.0 mM L-glutamine, 10% fetal bovine serum, antibiotics, and 0.05 IU/mL porcine follicle-stimulating hormone (FSH+) or vehicle control (CTL) medium for 22 h. RESULTS: FSH withdrawal (CTL) diminished several processes associated with the acquisition of oocyte developmental competence, such as reduced cumulus cell expansion, diminished estradiol synthesis (FSH+: 116.0±0.0 pg/mL vs. CTL: 97.6±18.0 pg/mL), and lower oocyte nuclear maturation rate (FSH+: 96.47% vs. CTL: 88.76%). Fresh media formulations (i.e., TCM199 with FSH or vehicle) were indistinguishable under biochemical profiling threshold conditions. Biochemical profiling showed similar total protein and lipid concentrations between groups. Further, total sugar concentrations diminished from fresh media to their post-IVM counterparts, albeit in an FSH-independent manner. Glycogen concentrations remained unaltered after IVM within CTL media, albeit were substantially lower after IVM under FSH+ conditions. CONCLUSION: FSH mediates the consumption of serum-derived glycogen by bovine COCs during IVM and implies that serum-free media should contain increased glucose concentrations to facilitate the acquisition of oocyte developmental competence.

Immunohistochemical and histophysiological study of prolonged use of nandrolone on reproductive organs and fertility.

We investigated possible changes in morphology and immunohistochemistry of the uterus and ovaries of rats caused by nandrolone (ND); we also investigated effects on fertility. We used 30 rats divided into three experimental groups: control (C), control vehicle (CV) and 5 mg/kg ND. Rats treated with ND exhibited loss of estrous cyclicity with predominance of the estrus phase, increased body weight and an organosomatic index that was decreased for the ovaries, but increased for the uterus. In the ovary, we observed a reduction in primary and secondary follicles and an increase in tertiary follicles; no corpora lutea were observed. Estrogen and progesterone levels were reduced. In the uterus, the endometrium was edematous with hyperplasic glands. The cytokines, TNFα and IL6, and the apoptotic index were increased in rats treated with ND. VEGF-A was increased in the ovaries and decreased in the uterus. We conclude that ND disrupts ovarian and uterine histophysiology by establishing an anovulatory and inflammatory condition, which directly affects reproduction.

Essential oil toxicity on biological and reproductive parameters of Alabama argillacea (Hübner) (Lepidoptera: Erebidae).

Several studies have demonstrated the effects of essential oils on insect pests. These effects vary and affect fundamental parameters for the survival of these organisms. However, there is a scarcity of research on the effect of these substances on Alabama argillacea (Hübner) (Lepidoptera: Erebidae), the main defoliating pest of cotton (Gossypium hirsutum L.). Thus, the present study aimed to evaluate the activity of essential oils from Litsea cubeba (Lour.) Pers., Melaleuca alternifolia Cheel, Juniperus virginiana L., and Mentha spicata L., on the biological and reproductive parameters of A. argillacea and the consequent effects on the gonads of both males and females of this pest. All essential oils presented toxicity by contact in third instar larvae of A. argillacea, causing a reduction in the weight of larvae and pupae, as well as affecting the number and viability of eggs. These essential oils changed the histochemistry of the testicles, and M. alternifolia, J. virginiana, and M. spicata also affected their morphology. The histochemistry of the ovarioles was altered by the essential oils from M. alternifolia, J. virginiana, and L. cubeba. Thus, the essential oils tested in the present work are promising for the control of A. argillacea, as they are toxic and affect the development and reproduction of this key pest of cotton.

Histological, histochemical and energy disorders caused by R-limonene on Aedes aegypti L. larvae (Diptera: Culicidae).

The study evaluated the effects of R-limonene, in sublethal concentration, on the histology, histochemistry, biochemistry, and carbohydrates and proteins levels in the third instar Aedes aegypti larvae. The R-limonene (LC50 of 27 ppm) and control groups were analyzed 12 and 24 h after the beginning of treatments. The midgut of the control larvae was composed of cylindrical and elongated cells with a spherical and central nucleus and regenerative cells with a pyramidal shape. After 12 h of treatment, columnar cells, protuberances, and cytoplasmic vacuolization were found. However, 24 h after treatment, complete disorganization of the epithelium was observed. There was a positive reaction in all treatments for the presence of glycogen. However, the midgut of larvae treated with R-limonene showed higher levels. For the total protein, positive marking occurred in all groups evaluated, with higher levels in treatments and the lowest in control 12 h. The levels of total protein and glycogen increased in the treated larvae compared to the 12 h control. Besides, a reduction in total sugar levels was observed in the treated larvae compared to controls 12 and 24 h, being more evident in the last one. Therefore, these results demonstrate that R-Limonene caused pathological changes in the epithelium of the A. aegypti midgut at histophysiological and biochemical levels.

Renal and hepatic changes in the offspring of rats that received biological insecticides during pregnancy and lactation.

Bacillus thuringiensis insecticides have been considered safe, being an alternative to the use of synthetic insecticides. However, studies have shown the effects of Bt Cry toxins on various organs, compromising their functions. The objective of this work was to test whether the administration of biological insecticides based on B. thuringiensis in pregnant rats will cause histopathological changes in the liver and kidneys, as well as in the levels of toxicity biomarkers, of their puppies in adulthood. Twenty rats, 90 days old, were used, divided into four groups: GC - Pregnant rats, GX - Pregnant rats that received XenTari®, GDi - Pregnant rats that received Dipel® and GDe - Pregnant rats that received deltamethrin. Insecticides were administered by gavage at a dosage of 1 mg/100 g/day (GX and GDi), and 2 mg/Kg/day (GDe) during pregnancy and lactation. In the animals of the groups whose matrices received the insecticides, there was a reduction in the levels of the biomarkers of toxicity alanine aminotransferase (ALT), aspartate aminotransferase (AST), urea and creatinine, about the control group. The biological insecticides promoted histopathological changes in the liver, with the presence of portal vein, centrilobular and sinusoidal capillaries congestion, and in the kidney, presence of cortical congestion and reduction of the subcapsular space. Histochemical evaluation in the liver demonstrated a significant reduction in glycogen in the groups that received insecticides when compared to the control group, whereas for collagen fibers in both the liver and the kidneys, no differences were observed between the experimental groups. The morphometry of the liver revealed a significant reduction in the lobular parenchyma and an increase in the non-lobular parenchyma, and in the kidney, there was a reduction in the diameter and volume of the glomerulus and Bowman's capsule of the animals whose matrices received both biological and synthetic insecticides. Thus, it is concluded that the biological insecticides XenTari® and Dipel® in sublethal doses in pregnant rats promote changes in the liver and kidney of the offspring similar to the insecticide deltamethrin, which extend into adulthood.

Effect of melatonin on gonad and thyroid development of offspring of hypothyroid pregnant rats.

We investigated the effects of melatonin on rats with induced hypothyroidism during gestation as well as its effect on the development of the gonads of their offspring. Fifteen pregnant rats were divided into three groups: GC, rats without induced hypothyroidism; GH, rats with induced hypothyroidism; GHM, rats with induced hypothyroidism plus melatonin. Hypothyroidism was induced by oral administration of 6-propyl-2-thiouracil and melatonin was applied subcutaneously. Treatments were performed during gestation and lactation. For the matrices, we evaluated the number of pups, body weight gain, ovarian weight, thyroid weight, organosomatic index, thyroid stimulating hormone (TSH) dose and thyroid morphometry. For the pups, weight gain, TSH, weight, morphometry of the gonads and organosomatic index were analyzed, as well as the cell proliferation index. TSH was elevated only in the matrices of GH animals. Melatonin prevented reduction of ovarian and thyroid weight, number of pups, follicular diameter and thyroid epithelial proportion of the matrices with hypothyroidism. The offspring of rats of the GH group exhibited less body weight gain, gonad and thyroid weight, and gonad cell proliferation index compared to the offspring born of rats of the GC and GHM groups. Melatonin prevented the effects of maternal hypothyroidism on the offspring of rats.

Effect of essential oils of Mentha spicata L. and Melaleuca alternifolia Cheel on the midgut of Podisus nigrispinus (Dallas) (Hemiptera: Pentatomidae).

Agricultural pest control is a popular research topic, and essential oils are widely studied because they represent a promising alternative to synthetic insecticides. However, despite the increase in studies on pests, little work has been done on pesticide contamination of the predators feeding on insecticide-affected prey. Therefore, the objective of this study was to evaluate the effects of Lethal Dose 50 (LD50) from the essential oils of Mentha spicata L. and Melaleuca alternifolia Cheel on the histology, including histochemistry (from protein and carbohydrate analysis) and immunohistochemistry (from the evaluation of cellular apoptosis), of the midgut of fifth instar nymphs of Podisus nigrispinus (stinkbug) (Dallas). The periods of analysis were 12, 24, and 48 h after ingestion of Alabama argillacea Hübner caterpillars treated with the respective oils. The oil from M. spicata did not cause histological alterations or apoptosis in the insect. However, there was a reduction in the level of carbohydrates within the 48-h period. After 24 h, the oil of M. alternifolia caused an elongation of digestive cells and, after 48 h, cell lysis with the release of material into the lumen, suggesting tissue necrosis. The immunohistochemical study revealed no apoptotic process. There was a reduction in the neutral carbohydrate levels in the 24- and 48-h periods and in the number of regenerative cells, when compared to the control, after the period of 48 h. These results demonstrate that M. spicata oil has potential for use in cotton fields because it does not affect the vital characteristics of P. nigrispinus. However, the essential oil of M. alternifolia is not suitable for use as a pesticide because it is extremely toxic to predators.

Protective action of melatonin on diabetic rat testis at cellular, hormonal and immunohistochemical levels.

The aim of the present study was to analyze the effects of melatonin treatment on diabetic rat testes. Fifty albino rats were divided into the following groups: CG: control group; GD: placebo-induced and placebo-treated mice; GDI: insulin-induced and post-confirmation diabetes-induced rats; GDM: diabetes-induced and melatonin-treated post-confirmation mice and GDMS: diabetes-induced and melatonin-treated mice simultaneously. Melatonin was administered at a dose of 10 mg/kg in drinking water every day for 20 days at night. Diabetes was induced by streptozotocin (60 mg/kg) and confirmed after the fifth day of induction. Insulin was administered at 5 IU (international units)/day at different times of the day for 20 days. The testes were submitted to histopathological, morphometric, immunohistochemical and oxidative stress analysis. Melatonin moderately decreased glycemic levels, protected weight loss and morphometric changes in the testicles, increased antioxidant enzyme levels and stabilized plasma testosterone and androgen receptor levels and decreased inflammatory markers in the testicles. Showing its potential to mitigate diabetes effects.

Ultrastructure of Tuta absoluta parasitized eggs and the reproductive potential of females after parasitism by Metarhizium anisopliae.

Among the microorganisms used in biological control, the muscadine fungus Metarhizium anisopliae (Metsch.) Sorok. is produced and formulated world wide aiming to control pests from several agricultural crops. This work evaluated effects of M. anisopliae isolate UFRPE-6 on the fecundity and mortality of Tuta absoluta (Lepidoptera: Gelechiidae) females and the mechanism of infection on eggs. The infection of the females by the fungus did not affect their oviposition and fecundity; however it affected the survival with total and confirmed mortality of 54.2% and 37.14%, respectively. The eggs were treated with suspension at concentration of 10(6)conidia/mL. The analysis under scanning electron microscopy showed that conidia germination and penetration processes in the eggs of T. absoluta started within the period of 6h after the inoculation. Several hyphal bodies were observed from 12h and an intense extrusion of the mycelium covering all the external surface of the eggs 72h after inoculation. Despite its moderate activity in adults, the isolate URPE-6 of M. anisopliae showed promising in the control of T. absoluta due to its pathogenicity and virulence to eggs from this pest.

Composition and Toxicity of a Mixture of Essential Oils Against Mediterranean Fruit Fly, Ceratitis capitata (Wiedemann) (Diptera: Tephritidae).

A natural insecticide developed from the mixture of the essential oils (EOs) of Cymbopogon citratus (DC.) Stapf (Poaceae), Cedrus atlantica (Endl.) Manetti ex Carriére (Pinaceae), and Corymbia citriodora (Hook.) K.D. Hill & L.A.S. Johnson (Myrtaceae) was studied. The mixture of oils caused high mortality (LD50 = 0.018 µl/insect) to the Mediterranean fruit fly, Ceratitis capitata (Wiedemann) (Diptera: Tephritidae), a globally important pest, after topical application on adults. Based on the chemical characterization of biopesticide using gas chromatography-mass spectrometry (GC-MS) analysis, many of the constituents had known insecticidal properties (the monoterpenes α-citronellal and E-citral and the sesquiterpernes α-himachalene and ß-himachalene, all at concentrations above 15%). Phytotoxicity tests on orange trees demonstrated that this natural product was harmless when the mixture was applied diluted in water with a surfactant. The mixture of EOs was also harmless (1, IOBC category) to the biological control agent responsible for reducing populations of tephritids, the parasitoid Psyttalia concolor (Hymenoptera: Braconidae), following exposure to treated orange trees in a semifield assay within a greenhouse, but killed 46.2% of C. capitata (Szépligeti) adults after 72 h. Our results suggest that mixture of EOs has potential for use as an adulticide against medfly, although the production price was exceedingly high compared with that of commercial synthetic insecticides. Therefore, we discuss the advantages and disadvantages related to the potential use of this natural insecticide.