Evaluating Volatile Plant Compounds of Psidium galapageium (Myrtales: Myrtaceae) as Repellents Against Invasive Parasitic Diptera in the Galapagos Islands.

Plant-based repellents represent a safe, economic, and viable alternative to managing invasive insects that threaten native fauna. Observations of self-medication in animals can provide important cues to the medicinal properties of plants. A recent study in the Galapagos Islands found that Darwin's finches apply the leaves of Psidium galapageium (Hooker 1847) to their feathers, extracts of which were repellent to mosquitoes and the parasitic fly Philornis downsi (Dodge & Aitkens 1968; Diptera: Muscidae). Introduced mosquitoes are suspected vectors of avian pathogens in the Galapagos Islands, whereas the larvae of P. downsi are blood-feeders, causing significant declines of the endemic avifauna. In this study, we investigated the volatile compounds found in P. galapageium, testing each against a model organism, the mosquito Anopheles arabiensis (Patton 1905; Diptera: Culicidae), with the aim of singling out the most effective compound for repelling dipterans. Examinations of an ethanolic extract of P. galapageium, its essential oil and each of their respective fractions, revealed a mixture of monoterpenes and sesquiterpenes, the latter consisting mainly of guaiol, trans-nerolidol, and ß-eudesmol. Of these, trans-nerolidol was identified as the most effective repellent to mosquitoes. This was subsequently tested at four different concentrations against P. downsi, but we did not find a repellence response. A tendency to avoid the compound was observed, albeit significance was not achieved in any case. The lack of repellence suggests that flies may respond to a combination of the volatile compounds found in P. galapageium, rather than to a single compound.

Mechanism of Action of Endophytic Fungi Hypocrea lixii and Beauveria bassiana in Phaseolus vulgaris as Biopesticides against Pea Leafminer and Fall Armyworm.

Endophytic fungal isolates Hypocrea lixii F3ST1 and Beauveria bassiana G1LU3 were evaluated for their potential to endophytically colonize and induce active compounds in Phaseolus vulgaris, as a defense mechanism against pea leafminer (Liriomyza huidobrensis) and fall armyworm (Spodoptera frugiperda). Endophytic colonization was achieved through seed inoculation with the volatile emissions from P. vulgaris plants being analyzed using GC-MS. The crude extracts of P. vulgaris obtained using methanol and dichloromethane were assayed against leafminer and fall armyworm larvae using leaf dipping and topical application, respectively. The two isolates successfully colonized the entire host plant (roots, stems, and leaves) with significant variation (p < 0.001) between fungal isolates and the controls. The results showed qualitative differences in the volatile profiles between the control plants, endophytically colonized and insect-damaged plants attributed to fungal inoculation and leafminer damage. The crude methanol extracts significantly reduced the percentage pupation of 2nd instar leafminer larvae (p < 0.001) and adult-flies emergence (p < 0.05). The survival of the 1st instar fall armyworm larvae was also significantly reduced (p < 0.001) compared to the controls. This study demonstrated the high potential of endophytic fungi H. lixii and B. bassiana in inducing mainly specific defense compounds in the common bean P. vulgaris that can be used against pea leafminer and fall armyworm.

Occurrence of Antibiotic Resistance Genes in Hermetia illucens Larvae Fed Coffee Silverskin Enriched with Schizochytrium limacinum or Isochrysis galbana Microalgae.

Hermetia illucens larvae are among the most promising insects for use as food or feed ingredients due to their ability to convert organic waste into biomass with high-quality proteins. In this novel food or feed source, the absence of antibiotic-resistant bacteria and their antibiotic resistance (AR) genes, which could be horizontally transferred to animal or human pathogens through the food chain, must be guaranteed. This study was conducted to enhance the extremely scarce knowledge on the occurrence of AR genes conferring resistance to the main classes of antibiotics in a rearing chain of H. illucens larvae and how they were affected by rearing substrates based on coffee silverskin supplemented with increasing percentages of Schizochytrium limacinum or Isochrysis galbana microalgae. Overall, the PCR and nested PCR assays showed a high prevalence of tetracycline resistance genes. No significant effect of rearing substrates on the distribution of the AR genes in the H. illucens larvae was observed. In contrast, the frass samples were characterized by a significant accumulation of AR genes, and this phenomenon was particularly evident for the samples collected after rearing H. illucens larvae on substrates supplemented with high percentages (>20%) of I. galbana. The latter finding indicates potential safety concerns in reusing frass in agriculture.

Effect of Homeopathic Medicines and a Nosode on Larvae of Cochliomyia hominivorax (Diptera: Calliphoridae).

BACKGROUND: Cochliomyia hominivorax is the major fly causing primary myiasis in livestock animals in Brazil; its larvae develop in the host's living tissues, causing mutilations, which can even lead to death. In conventional treatments of myiasis, chemo-synthetic insecticides have been employed directly on larvae present in the wounds. Homeopathy may represent a healthy and sustainable alternative both to prevent and to treat myiasis in animals and humans. The current study evaluated how the emergence of adult insects is affected by the use of the homeopathic medicines Sulfur 12cH and Pyrogenium 12cH, and the nosode produced from C. hominivorax larvae at potencies 8cH and 12cH, on third-stage larvae of a C. hominivorax colony. MATERIALS AND METHODS: The homeopathic medicines and the nosodes were produced according to the Brazilian Homeopathic Pharmacopoeia. Control groups were distilled water, alcohol, no substance, and the organophosphate insecticide trichlorfon. For each group, 10 replicates were performed. Emergence rate of adult insects was evaluated by descriptive statistics followed by analysis of variance. Homogeneity of variances was verified by F-test and group means were compared with Tukey's test. RESULTS: Mortality rates in control groups were 2.7% for 30% (v/v) alcohol, 4.3% for distilled water, 3.2% for no substance (p > 0.05). In the trichlorfon group, the mortality rate of larvae was 90.8%. For Sulfur 12cH, the mortality of larvae was 94.6%, and for Pyrogenium 12cH it was 98.6%. The latter three means were not statistically different from each other or from the mean found for the trichlorfon group. The mortality rates of larvae were 61.3% and 66.6% for nosode C. hominivorax 8cH and 12cH, respectively (p > 0.05). CONCLUSIONS: Results suggest that homeopathy could be used therapeutically to prevent and treat animals and humans with myiasis caused by C. hominivorax.

Toxicity and oviposition deterrence of essential oils of Clinopodium nubigenum and Lavandula angustifolia against the myiasis-inducing blowfly Lucilia sericata.

Cutaneous myiasis is a severe worldwide medical and veterinary issue. In this trial the essential oil (EO) of the Andean medicinal plant species Clinopodium nubigenum (Kunth) Kuntze was evaluated for its bioactivity against the myiasis-inducing blowfly Lucilia sericata (Meigen) (Diptera Calliphoridae) and compared with that of the well-known medicinal plant species Lavandula angustifolia Mill. The EOs were analysed and tested in laboratory for their oviposition deterrence and toxicity against L. sericata adults. The physiology of EO toxicity was evaluated by enzymatic inhibition tests. The antibacterial and antifungal properties of the EOs were tested as well. At 0.8 µL cm-2, both EOs completely deterred L. sericata oviposition up to 3 hours. After 24 h, the oviposition deterrence was still 82.7% for L. angustifolia and the 89.5% for C. nubigenum. The two EOs were also toxic to eggs and adults of L. sericata. By contact/fumigation, the EOs, the LC50 values against the eggs were 0.07 and 0.48 µL cm-2 while, by topical application on the adults, LD50 values were 0.278 and 0.393 µL per individual for C. nubigenum and L. angustifolia EOs, respectively. Inhibition of acetylcholine esterase of L. sericata by EOs (IC50 = 67.450 and 79.495 mg L-1 for C. nubigenum and L. angustifolia, respectively) suggested that the neural sites are targets of the EO toxicity. Finally, the observed antibacterial and antifungal properties of C. nubigenum and L. angustifolia EOs suggest that they could also help prevent secondary infections.

Tissue damage and cytotoxic effects of Tagetes minuta essential oil against Lucilia cuprina.

The blowfly Lucilia cuprina has great medico-sanitary and veterinary importance due to the ability of its larval form to develop in decaying organic matter, parasitizing vertebrates. Fly eradication is challenging and the essential oil (EO) of Tagetes minuta (TMEO) have been reported to have therapeutic properties. This study aimed to determine the activity of EO from the aerial parts of T. minuta against third instar larvae (L3) of L. cuprina. Groups of 20 L3 were placed on filter paper, which were impregnated with varying concentrations (0.19; 0.39; 0.79; 1.59; 2.38; 3.18; 4.77; and 6.36 µL/cm2) of TMEO solubilized in acetone, ethanol or Tween 20. Histological tissue damage of TMEO was measured in L3 after 24, 48 and 96 h of exposure. Dihydrotagetone (67.64%), trans-ocimene (16.23%), trans-tagetone (10.14%) and verbenone (2.98%) were obtained as major compounds of TMEO. Lethal concentrations of 50%, 24 and 48 h after TMEO exposure were 1.02 and 0.73 µL/cm2 for acetone; 3.37 and 1.75 µL/cm2 for ethanol; and 7.46 and 6.11 µL/cm2 for Tween 20, respectively. TMEO had a significant L3 mortality of 96.6% in acetone, 48 h after contact. Cuticle abnormalities were observed, as well as the loss of digestive tract architecture and vacuolization in fat bodies. TMEO presented time and concentration-dependent effects against L. cuprina. As our study demonstrated a strong insecticide activity of TMEO, we consider that it could be developed into an ecofriendly product against L. cuprina.

Ovicidal and Deleterious Effects of Cashew (Anacardium occidentale) Nut Shell Oil and Its Fractions on Musca domestica, Chrysomya megacephala, Anticarsia gemmatalis and Spodoptera frugiperda.

In this work, we evaluated the ovicidal activity and the deleterious effects of cashew (Anacardium occidentale) nut shell oil and its fractions on the development of Musca domestica and Chrysomya megacephala, important vectors of several diseases. The insecticidal effects of this plant were also measured on the first and second instar larvae of Anticarsia gemmatalis and Spodoptera frugiperda, soy and maize pests, respectively. The fly eggs and the crop pest insect larvae were exposed to the cashew (Anacardium occidentale) nut shell liquid (CNSL) and its fractions: technical CNSL, anacardic acid, cardanol and cardol. The results show that the cardol fraction, for both species of flies, presented the lowest lethal concentration with LC50 of 80.4 mg/L for M. domestica and 90.2 mg/L for C. megacephala. For the mortality of the larvae of A. gemmatalis and S. frugiperda, the most effective fraction was anacardic acid with LC50 of 295.1 mg/L and 318.4 mg/L, respectively. In all species, the mortality rate of the commercial compounds (cypermethrin 600 mg/L and temephos 2 mg/L) was higher than that of the evaluated compounds. Despite this, the results obtained suggest their potential in field trials, once the fractions of A. occidentale presented high mortality at low lethal concentrations in laboratory conditions, with the possibility of integrated use in the control of disease vectors and agricultural pests, employing ecofriendly compounds.

Potential of essential oils to prevent fly strike and their effects on the longevity of adult Lucilia sericata.

Lucilia sericata is a facultative ectoparasite causing fly strike or myiasis in warm-blooded vertebrates. It is controlled by traps or insecticides, but both have drawbacks and alternative ways of control are urgently needed. Essential oils (EOs) of vetiver (Chrysopogon zizanioides), cinnamon (Cinnamomum zeylanicum), and lavender (Lavandula angustifolia) and their blends (OBs); OB1 (2 ml of each EO plus 4 ml of sunflower oil as a carrier) and OB2 (2 ml of each EO) were tested. Oils were tested at 5% for deterrence assays, and a dose response assay 0.01-0.6%, was conducted to determine forced-contact toxicity. We evaluated the efficacy of oils as oviposition deterrents, repellents/attractants, and their effects on mortality and longevity of adult L. sericata. Our data indicated that 0.2% EOs killed all flies by 5 min post-treatment and that vetiver oil greatly deterred flies from the oviposition medium and reduced adult longevity. Sunflower oil repelled all flies from ovipositing and greatly reduced the lifespan of treated adults. The blend of the four oils (OB1) had the greatest repellent effect on the flies. EOs have insecticidal, repellent, and oviposition-deterrent activities against L. sericata that could be used for suppression of blow fly populations.

The chemical structure and anti-aging bioactivity of an acid polysaccharide obtained from rose buds.

An acid polysaccharide, named R-PL, was extracted from rose buds by hot water (80 °C) extraction and purified by Sephadex G-200. R-PL, extracted with a total yield of 1.90%, is a highly pure polysaccharide with a total sugar content of 92.51%. A single and symmetrically sharp peak in its high-performance gel-permeation chromatography (HPGPC) spectrum indicates that R-PL is a homogeneous polysaccharide with a molecular weight of 7.727 × 105 Da. Monosaccharide composition analysis shows that it consists of d-arabinose, d-xylose, d-glucose, d-galactose, d-galacturonic acid, and d-glucuronic acid with a molar ratio of 4.6 : 1.4 : 5.22 : 4.81 : 1 : 1.86. Structure analysis, such as FTIR spectroscopy, periodic acid oxidation, Smith degradation, methylation, and NMR analysis, reveals that R-PL has a backbone of →1)-α-d-Glcp-(6→ with side chains of →1)-ß-d-Galp-(4,6→, →1)-ß-d-Araf-(5→, →1)-α-d-Xylp-(4→, →1)-α-d-GlcpA-(2→ and →1)-α-d-GalpA-(4→. Its morphological characteristics were identified by Congo red experiments and scanning electron microscopy (SEM) analysis. The results show that R-PL does not have a triple-helical conformation in solution; its shape resembles twisted ribbons. Anti-oxidative and anti-aging analyses show that R-PL has significant antioxidant and anti-aging abilities in vivo.

Ovicidal and larvicidal activity and possible mode of action of phenylpropanoids and ketone identified in Syzygium aromaticum bud against Bradysia procera.

Bradysia procera is a serious insect pest of Panax ginseng plants. This study was conducted to determine the toxicity and mechanism of action of three phenylpropanoids, three terpenoids, and a ketone from Syzygium aromaticum bud methanol extract and hydrodistillate against third-instar larvae and eggs of B. procera. In a filter-paper mortality bioassay, methyl salicylate (LC50, 5.26µg/cm2) was the most toxic compound, followed by 2-nonanone, eugenol, and eugenyl acetate (8.77-15.40µg/cm2). These compounds were significantly less toxic than either thiamethoxam, clothianidin, or cypermethrin. Egg hatching was inhibited by 97, 85, and 40% at 11.7µg/cm2 of methyl salicylate, 2-nonanone, and eugenol, respectively. The egg-hatching inhibition of these insecticides was between 90 and 94% at 0.09µg/cm2. These constituents were consistently more toxic in closed versus open containers, indicating that toxicity was achieved mainly through the action of vapor. The mechanism of larvicidal action of methyl salicylate, eugenol, and eugenyl acetate might be primarily due to interference with the octopaminergic system. 2-Heptyl acetate and 2-nonanone might act on both acetylcholinesterase and the octopaminergic receptor. 2-Heptanone might act primarily on acetylcholinesterase. Further studies will warrant possible applications of S. aromaticum bud-derived products as potential larvicides and ovicides for the control of B. procera.

Toxicity and growth inhibition potential of vetiver, cinnamon, and lavender essential oils and their blends against larvae of the sheep blowfly, Lucilia sericata.

BACKGROUND: Myiasis induced by the sheep blowfly, Lucilia sericata, represents a public health problem widely distributed throughout the world. L. sericata larval stages feed on both humans and animals. L. sericata adults and larvae can play a role in spreading agents of mycobacterial infections. OBJECTIVES: It is critical to establish new and safe alternative methods of controlling L. sericata. METHODS: The insecticidal effectiveness and growth inhibition potential of three commercially available essential oils (EOs), vetiver (Chrysopogon zizanioides), cinnamon (Cinnamomum zeylanicum), and lavender (Lavandula angustifolia), as well as their blends, were tested against the second (L2) and third (L3) larval stages of L. sericata. Sunflower (Helianthus annuus) oil was used as a carrier and tested on L2 and L3 larvae. To the best of our knowledge, all applied essential oils, except lavender, and oil blends were tested against L. sericata for the first time. RESULTS: All applied oils did not repel L2 from the treated liver but adversely affected their development. Contact treatments on L. sericata L3 indicated that vetiver and cinnamon oils significantly affected treated larvae. Total mortality rates were 93.33 and 95.56%, respectively. Furthermore, oil blends tested through contact assays killed larvae when used at higher concentrations; adult emergence was eliminated post-treatment with doses >30% for oil blend 1 and >10% for oil blend 2. CONCLUSION: Overall, cinnamon and vetiver oils (5%) were selected as reliable and cheap biopesticides for controlling larvae of L. sericata. The tested oils are inexpensive and represent new promising botanical insecticides in the fight against blowflies causing myiasis.

Insecticide activity of Baccharis dracunculifolia essential oil against Cochliomyia macellaria (Diptera: Calliphoridae).

The ethnobotanical uses of Brazilian plants for different injuries and diseases conjoined with local rich biodiversity represent an important resource for research and development. This study aimed to characterise BDEO and its in vitro activity on the third instar larvae (L3) of Cochliomyia macellaria. Groups of 20 L3 were placed on filter paper impregnated with increasing concentrations of 5-30% (v/v), equivalent to 0.79-4.77 µL/cm2, solubilised in ethanol or acetone. The major constituents of BDEO were ß-pinene (9.94%), D-limonene (9.59%), ß-nerolidol (7.93%), caryophyllene (7.69%), spathulenol (6.69), α-muurolene (6.74%) and α-pinene (5.31%). Lethal concentrations of 50% for BDEO on C. macellaria (LC50) after 24 and 48 h of exposure were 2.63 and 2.47 µL/cm2 for ethanol and 9.58 and 8.11 µL/cm2 for acetone, respectively. Furthermore, larvae cuticle abnormalities and adult deformity were observed. Our data confirm the effectiveness of BDEO as an ecofriendly product against blowflies.

Artemisia spp. essential oils against the disease-carrying blowfly Calliphora vomitoria.

BACKGROUND: Synanthropic flies play a considerable role in the transmission of pathogenic and non-pathogenic microorganisms. In this work, the essential oil (EO) of two aromatic plants, Artemisia annua and Artemisia dracunculus, were evaluated for their abilities to control the blowfly Calliphora vomitoria. Artemisia annua and A. dracunculus EOs were extracted, analysed and tested in laboratory bioassays. Besides, the physiology of EOs toxicity and the EOs antibacterial and antifungal properties were evaluated. RESULTS: Both Artemisia EOs deterred C. vomitoria oviposition on fresh beef meat. At 0.05 µl cm-2 A. dracunculus EO completely inhibited C. vomitoria oviposition. Toxicity tests, by contact, showed LD50 of 0.49 and 0.79 µl EO per fly for A. dracunculus and A. annua, respectively. By fumigation, LC50 values were 49.55 and 88.09 µl l-1 air for A. dracunculus and A. annua, respectively. EOs AChE inhibition in C. vomitoria (IC50 = 202.6 and 472.4 mg l-1, respectively, for A. dracunculus and A. annua) indicated that insect neural sites are targeted by the EOs toxicity. Finally, the antibacterial and antifungal activities of the two Artemisia EOs may assist in the reduction of transmission of microbial infections/contaminations. CONCLUSIONS: Results suggest that Artemisia EOs could be of use in the control of C. vomitoria, a common vector of pathogenic microorganisms and agent of human and animal cutaneous myiasis. The prevention of pathogenic and parasitic infections is a priority for human and animal health. The Artemisia EOs could represent an eco-friendly, low-cost alternative to synthetic repellents and insecticides to fight synanthropic disease-carrying blowflies.

Telipogon peruvianus (Orchidaceae) Flowers Elicit Pre-Mating Behaviour in Eudejeania (Tachinidae) Males for Pollination.

Several neotropical orchid genera have been proposed as being sexually deceptive; however, this has been carefully tested in only a few cases. The genus Telipogon has long been assumed to be pollinated by male tachinid flies during pseudocopulatory events but no detailed confirmatory reports are available. Here, we have used an array of methods to elucidate the pollination mechanism in Telipogon peruvianus. The species presents flowers that have a mean floral longevity of 33 days and that are self-compatible, although spontaneous self-pollination does not occur. The flowers attract males of four tachinid species but only the males of an undescribed Eudejeania (Eudejeania aff. browni; Tachinidae) species are specific pollinators. Males visit the flowers during the first few hours of the day and the pollination success is very high (42% in one patch) compared with other sexually deceptive species. Female-seeking males are attracted to the flowers but do not attempt copulation with the flowers, as is usually described in sexually deceptive species. Nevertheless, morphological analysis and behavioural tests have shown an imperfect mimicry between flowers and females suggesting that the attractant stimulus is not based only on visual cues, as long thought. Challenging previous conclusions, our chemical analysis has confirmed that flowers of Telipogon release volatile compounds; however, the role of these volatiles in pollinator behaviour remains to be established. Pollinator behaviour and histological analyses indicate that Telipogon flowers possess scent-producing structures throughout the corolla. Our study provides the first confirmed case of (i) a sexually deceptive species in the Onciidinae, (ii) pollination by pre-copulatory behaviour and (iii) pollination by sexual deception involving tachinid flies.

Molecular basis of the remarkable species selectivity of an insecticidal sodium channel toxin from the African spider Augacephalus ezendami.

The inexorable decline in the armament of registered chemical insecticides has stimulated research into environmentally-friendly alternatives. Insecticidal spider-venom peptides are promising candidates for bioinsecticide development but it is challenging to find peptides that are specific for targeted pests. In the present study, we isolated an insecticidal peptide (Ae1a) from venom of the African spider Augacephalus ezendami (family Theraphosidae). Injection of Ae1a into sheep blowflies (Lucilia cuprina) induced rapid but reversible paralysis. In striking contrast, Ae1a was lethal to closely related fruit flies (Drosophila melanogaster) but induced no adverse effects in the recalcitrant lepidopteran pest Helicoverpa armigera. Electrophysiological experiments revealed that Ae1a potently inhibits the voltage-gated sodium channel BgNaV1 from the German cockroach Blattella germanica by shifting the threshold for channel activation to more depolarized potentials. In contrast, Ae1a failed to significantly affect sodium currents in dorsal unpaired median neurons from the American cockroach Periplaneta americana. We show that Ae1a interacts with the domain II voltage sensor and that sensitivity to the toxin is conferred by natural sequence variations in the S1-S2 loop of domain II. The phyletic specificity of Ae1a provides crucial information for development of sodium channel insecticides that target key insect pests without harming beneficial species.

An acetone extract of Clausena anisata may be a potential control agent for flies encountered in cutaneous myiasis.

Clausena anisata is a medicinal plant used traditionally to treat myiasis and as an insect repellent by various communities. We have previously demonstrated the effects of C. anisata extracts on blowfly feeding and development in our laboratory. The impact of C. anisata leaf extracts on populations of different fly species on farms in Mpumalanga, South Africa was investigated in this study under field conditions. Flies were exposed to liver baits treated with acetone leaf extracts of C. anisata (150 mg/mL). Fly numbers and composition on two farms, with and without C. anisata treated liver, were compared during a period of 12 weeks when fly populations were expected to be high. Observations were made on fly behaviour and development, adult sizes and numbers. The flies exposed to liver treated with the leaf extract of C. anisata had a decreased rate of development, prolonged larval period, smaller body sizes and more sluggish behaviour compared to those subjected to the control treatment. No significant differences were, however, found between the numbers and sizes of flies on the treated and on the control farm, which was most likely due to the limited nature of the baiting programme we followed. The effects of C. anisata extracts on blowfly behaviour and development observed in previous laboratory studies were confirmed in this field evaluation. Although the extracts did not have a significant effect on the overall population size in this experiment, we believe that the C. anisata leaf extract could be useful in integrated pest management based on its effect on larval development. In addition, species such as Lucilia cuprina and Chrysomya marginalis seemed to have been repelled by the C. anisata treated liver; as a result, further work should explore this aspect and how it can be used for the protection of animals.

Statin Concentrations Below the Minimum Inhibitory Concentration Attenuate the Virulence of Rhizopus oryzae.

BACKGROUND: Mucormycosis is a destructive invasive mold infection afflicting patients with diabetes and hematologic malignancies. Patients with diabetes are often treated with statins, which have been shown to have antifungal properties. We sought to examine the effects of statins on Rhizopus oryzae, a common cause of mucormycosis. METHODS: Clinical strains of R. oryzae were exposed to lovastatin, atorvastatin, and simvastatin and the minimum inhibitory concentrations (MICs) were determined. R. oryzae germination, DNA fragmentation, susceptibility to oxidative stress, and ability to damage endothelial cells were assessed. We further investigated the impact of exposure to lovastatin on the virulence of R. oryzae RESULTS: All statins had MICs of >64 µg/mL against R. oryzae Exposure of R. oryzae to statins decreased germling formation, induced DNA fragmentation, and attenuated damage to endothelial cells independently of the expression of GRP78 and CotH. Additionally, R. oryzae exposed to lovastatin showed macroscopic loss of melanin, yielded increased susceptibility to the oxidative agent peroxide, and had attenuated virulence in both fly and mouse models of mucormycosis. CONCLUSIONS: Exposure of R. oryzae to statins at concentrations below their MICs decreased virulence both in vitro and in vivo. Further investigation is warranted into the use of statins as adjunctive therapy in mucormycosis.

The impacts of larval density and protease inhibition on feeding in medicinal larvae of the greenbottle fly Lucilia sericata.

Larval therapy, the therapeutic use of blowfly larvae to treat chronic wounds, is primarily used in debridement. There are, however, gaps in current knowledge of the optimal clinical application of the therapy and mechanisms of action in the debridement process. Using an artificial assay, two studies were undertaken to investigate these aspects of larval debridement by Lucilia sericata Meigen (Diptera: Calliphoridae); the first studied the effects of the density of larvae on tissue digestion and larval mass, and the second considered the effects on the same parameters of incorporating protease inhibitors into the feeding substrate. The total mass of tissue digested increased with larval density until saturation was observed at 5.0-7.5 larvae/cm(2) . This range was considered optimal as lower doses resulted in the removal of less tissue and higher doses offered no additional tissue removal and appeared to exacerbate competition for feeding. In the second study, increased protease inhibitor concentration led to significant decreases in tissue digestion and larval mass, suggesting that serine proteases, particularly trypsin, may play major roles in larval digestion. Such information is important in elucidating the main constituents that make up larval digestive products and may be significant in the development of new therapies.

The bioefficacy of crude extracts of Azadirachta indica (Meliaceae) on the survival and development of myiasis-causing larvae of Chrysomya bezziana (Diptera: Calliphoridae).

Myiasis is a type of parasitosis originating from the invasion of tissues of live humans and other vertebrates by dipteran larvae. The Old World screwworm fly­Chrysomya bezziana­is known worldwide in the tropical regions for causing myiasis among man and domestic animals, thereby leading to health hazards and severe economic losses to the dairy farmers. Management techniques for controlling populations of the fly are needed to minimize these losses. Plant-derived materials have been increasingly evaluated these days in controlling the insects of medical and veterinary importance. This study evaluated the efficacy of crude extracts of the plant neem, Azadirachta indica, against C. bezziana. The dried leaves of the plant were extracted successively with four different solvents viz. petroleum ether, chloroform, ethyl acetate and methanol and were evaluated against the third instar larvae of C. bezziana using dipping method and thin film application technique. In the dipping method, larvae were dipped in four different concentrations of plant extracts for 30 s, whereas in the thin film application, they were exposed to a thin film of plant extracts. The results showed that all the extracts had toxic effect on the larvae in both the techniques. In the dipping method, the highest mortalities were recorded in methanol extract followed by chloroform, petroleum ether and ethyl acetate extracts with LC50 values 1.07 g/100 ml, 1.7 g/100 ml, 3.39 g/100 ml and 4.9 g/100 ml, respectively. In the thin film application method, methanol extract showed the highest mortalities followed by chloroform, ethyl acetate and petroleum ether with LC50 values 0.4 mg/cm2, 0.6 mg/cm2, 2.1 mg/cm2 and 2.5 mg/cm2. It is concluded that the crude extracts of A. indica can be used in controlling the larvae of C. bezziana by using the dipping as well as thin film application technique.

In vitro repellent effect of tea tree (Melaleuca alternifolia) and andiroba (Carapa guianensis) oils on Haemotobia irritans and Chrysomya megacephala flies.

This study aimed to evaluate the repellent effect of tea tree (Melaleuca alternifolia) and andiroba (Carapa guianensis) essential oils on two species of flies (Haemotobia irritans and Chrysomya megacephala). For the in vitro studies, free-living adult flies were captured and reared in the laboratory. To verify the repellency effect, an apparatus was constructed where H. irritans and C. megacephala were exposed to andiroba and tea tree oils (5.0%), as well as to a known repellent (citronella, 5.0%) to validate the test. The study demonstrated that all three oils used showed in vitro repellent effect against both species of flies. It is possible to conclude that the essential oils (tea tree and andiroba) have repellent effect on these species of flies used in this study.