A survey of mutations in the Cochliomyia hominivorax (Diptera: Calliphoridae) esterase E3 gene associated with organophosphate resistance and the molecular identification of mutant alleles

Cochliomyia hominivorax (Calliphoridae) is one of the most important myiasis-causing flies and is responsible for severe economic losses to the livestock industry throughout the Neotropical region. In Brazil, C. hominivorax has been controlled mainly with organophosphate (OP) insecticides, although the inappropriate use of these chemicals can result in the selection of resistant flies. Changes in carboxylesterase activity have been associated with OP insecticides in some arthopodan species. In this work, we isolated and characterized part of the E3 gene in C. hominivorax (ChE7), which contained the same substitutions responsible for the acquisition of OP hydrolase activity in Lucilia cuprina (Calliphoridae). Digestion of the polymerase chain reaction products with a restriction enzyme that specifically recognized the mutation site unambiguously differentiated wild and mutated esterase alleles. The PCR-RFLP assay therefore provided a fast, reliable DNA-based method for identifying C. hominivorax individuals with a mutation in the esterase gene. Further bioassays to determine the association of this mutation with OP resistance in C. hominivorax should allow the development of more effective strategies for managing this species.     

Trypanocidal Action of (?)-Elatol Involves an Oxidative Stress Triggered by Mitochondria Dysfunction

Natural compounds have shown good potential for the discovery of new chemotherapeutics for the treatment of Chagas’ disease. Recently, our group reported the effective trypanocidal activity of (−)-elatol, extracted from the red macroalgae Laurencia dendroidea present in the Brazilian coast against Trypanosoma cruzi. However, the mechanism of action of this compound has remained unclear. There are only hypotheses concerning its action on mitochondrial function. Here, we further investigated the mechanisms of action of (−)-elatol on trypomastigotes of T. cruzi. For this, we evaluated some biochemical alterations in trypomastigotes treated with (−)-elatol. Our results show that (−)-elatol induced depolarization of the mitochondrial membrane, an increase in the formation of mitochondrial superoxide anion and loss of cell membrane and DNA integrity. Additionally, (−)-elatol induced formation of autophagic vacuoles and a decrease in cell volume. All together, these results suggest that the trypanocidal action of (−)-elatol involves multiple events and mitochondria might be the initial target organelle. Our hypothesis is that the mitochondrial dysfunction leads to an increase of ROS production through the electron transport chain, which affects cell membrane and DNA integrity leading to different types of parasite death.     

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Structural development of Pacific red snapper Lutjanus peru from hatching to the onset of first feeding

Successful rearing of larval fish requires culture conditions and feeding strategies matching the ontogenetic status of larvae. This study describes the external morphology and development of organs and structures involved in the feeding process (i.e. sensorial organs, mouthparts and digestive system) from hatching until first feeding in Pacific red snapper. Hatching occurred 26 h after fertilization at 26°C and total length (TL) was 2.45 ± 0.08 mm. The larvae showed an undifferentiated eye and digestive tract. At 48 hah, TL was 3.31 ± 0.12 mm. Yolk and oil globule were still present. The mouth was still closed, but the Meckel's, quadrate, hyoid and hyomandibular cartilages were present. The retina was formed by 5 layers, and a thin layer of pigment epithelium was observed in the outer nuclear layer (ONL). At 70 hah, TL was 3.44 ± 0.22 mm. A remnant of oil globule was still present. The mouth and anus were open. At 93 hah, the number of cones in the ONL of the retina have increased and there was more pigment in the pigment epithelial layer. A joint between Meckel's and the quadrate cartilage and also a joint between the hyomandibular cartilage and the skull were present. The presence of live feed was detected in the digestive tract of these larvae. Based on these observations, the Pacific red snapper larvae is functional to start ingesting live feed between the 3rd and 4th day after hatching.     

Using Limestone to Improve Soil Fertility and Growth of Mango (Mangifera Indica L.)

Soil acidity is one of the most important factors limiting crop production. The objective of this work was to evaluate the effects of limestone application on the soil chemical properties, nutrition and yield of mango plants in an orchard under implementation. The design was randomized blocks, with five limestone doses (0; 2; 4; 6 and 8 t ha^?1) and four replications. Soil chemical analyses were performed (at 12, 24, 36 and 48 months after the experiment implementation) in the layers 0–20; 20–40 and 40–60 cm deep. Nutrition status and yield were assessed during the first and second crop seasons. The highest fruit yield was associated with the application of 3.9 and 3.8 t ha^?1 of limestone in the 2008/09 and 2009/10 seasons, respectively, that is, the dose recommended by the literature for raising base saturation to 80%, as a function of the fertility conditions of the soil initially obtained.     

Silage maize quality in different uses of Italian ryegrass and soil management methods after liming

The effects of lime application with multiple soil management methods and uses on dry matter (DM) yield and crude protein (CP) content of Silage maize cropping in succession with Italian ryegrass (ryegrass) from 2009 to 2014 in a southern Brazilian Oxisol were studied. The experimental design was completely randomized block in split-plot with four replications. The main plot treatments were the four soil management methods: conventional tillage (CT), minimum tillage (MT), no-tillage (NT) and chiseled no-tillage (CNT). The sub-plots treatments were the three uses of Italian ryegrass (ryegrass): cover crop (CC), silage (S), and grazing dairy heifers as part of integrated crop-livestock system (ICL). In all the years, the maize was sown approximately 30 days after the ryegrass desiccation with herbicide. In the medium-term (5 yrs.) after liming, soil management methods did not change DM yield and CP content in silage maize. The use of ryegrass for S and ICL did not change DM yield, but notably maintained or increased CP content in silage maize after liming. The use of ryegrass only as CC provided less measurable benefits than the combined production system of silage maize with ryegrass.     

Effect of provision of apiaceous flowers associated to foods on the biology of <Emphasis Type="Italic">Coleomegilla maculata</Emphasis>

Coleomegilla maculata DeGeer is an omnivorous lady beetle that feeds on natural or alternative prey and artificial foods, which allows its laboratory rearing for use on augmentative biocontrol. In addition, C. maculata may supplement its diet with pollen and nectar, which helps in its conservation in agroecosystems. This study aimed to evaluate if Apiaceous flowers (Anethum graveolens L. and Coriadrum sativum L.), with and without alternative prey [eggs of Ephestia kuehniella (Zeller) or larvae of Drosophila melanogaster Meigen] or an artificial food (aqueous solution of honey), may guarantee the survival and complete development of the immature and adult stages of C. maculata in the laboratory. The immature stages developed only when Apiaceous flowers were offered with E. kuehniella eggs. The food with only one of the alternative prey (moth eggs or fly larvae) or moth eggs + honey solution resulted in fertile adults; however, the number of eggs/cluster was greater for the foods with E. kuehniella eggs + honey solution, A. graveolens flowers, or only D. melanogaster. Foods comprising only the two Apiaceous species, only the honey solution, or only water resulted in larval development up to a specific instar. Adults of C. maculata also survived on these foods, but there was no oviposition. The foods of the two Apiaceous species produced heavier adults only when associated with E. kuehniella eggs. The results indicate that the zoophytophagous habit of C. maculata should be considered in conservation biocontrol programs aimed at using this lady beetle to control crop pests.     

Detection of <Emphasis Type="Italic">Brucella</Emphasis> sp. infection through serological,microbiological, and molecular methods applied to buffaloes in Maranhão State,Brazil

The aim of the current study is to diagnose Brucella spp. infection using methods such as serology, bacterial isolation, and molecular analysis in buffaloes bred in Maranhão State. In order to do so, 390 samples of buffalo serum were subjected to serological tests, to Rose Bengal Plate Test (RBPT) and to 2-mercaptoethanol (2-ME) combined with slow agglutination test (SAT). Vaginal swabs were collected from seropositive animals and subjected to bacterial isolation and to generic PCR. According to the serological test, 16 animals had a positive reaction to the confirmatory test (2-ME/SAT). As for bacterial isolation, three samples resulted in the isolation of Brucella spp.-characteristic colonies, which were confirmed through PCR. These results confirmed Brucella spp. infection in the buffalo herd from Maranhão State.     

Should the domestic buffalo (Bubalus bubalis) be considered in the epidemiology of Bovine Herpesvirus 1 infection?

Only limited information is available on the epidemiology and pathogenesis of Bovine Herpesvirus 1 (BoHV-1) in domestic buffalos. In this study, a virulent BoHV-1 field strain isolated from cattle was inoculated into buffaloes to evaluate their susceptibility to the virus and to investigate the establishment of viral latency through clinical, virological and serological investigations. Latency was also studied by attempting viral reactivation using pharmacological induction. Six of seven male, 5 months old buffaloes were intranasally inoculated with BoHV-1; the other animal was kept as negative control. The animals were clinically monitored during the post-infection (P.I.) and the post-pharmacological induction (P.P.) periods. During these periods, nasal and rectal swabs, and blood samples, with and without anticoagulant, were collected at 2–3 day intervals. On culling the animals, 206 days P.I., their trigeminal ganglia and tonsils were collected. No clinical signs referable to BoHV-1 were observed throughout the experimental period. However, seropositivity was detected in all infected animals within day 20 P.I., using BoHV-1 glycoprotein E and glycoprotein B competitive ELISAs (IDEXX) and virus neutralisation test. In real-time PCR (RT-PCR), five of these animals were positive, at least once, for nasal or rectal swabs, during the P.I. period. The sixth infected animal was found positive only in the trigeminal ganglia after culling. Ganglia were also positive for two other animals. Virus isolation in permissive cell-lines was successful for a part of the RT-PCR positive samples. The detected viruses were confirmed by genetic analysis as identical to the inoculated strain. No evidence of infection was observed in the negative control. This study represents the first experimental transmission of BoHV-1 in buffaloes, confirming their susceptibility to infection and their possible role as host/reservoirs of BoHV-1.     

Effects of methadone on the minimum alveolar concentration of isoflurane in dogs

ObjectiveTo investigate the effects of methadone on the minimum alveolar concentration of isoflurane (ISO_MAC) in dogs.Study designProspective, randomized cross-over experimental study.AnimalsSix adult mongrel dogs, four males and two females, weighing 22.8 ± 6.6 kg.MethodsAnimals were anesthetized with isoflurane and mechanically ventilated on three separate days, at least 1 week apart. Core temperature was maintained between 37.5 and 38.5 °C during ISO_MAC determinations. On each study day, ISO_MAC was determined using electrical stimulation of the antebrachium (50 V, 50 Hz, 10 mseconds) at 2.5 and 5 hours after intravenous injection of physiological saline (control) or one of two doses of methadone (0.5 or 1.0 mg kg^?1).ResultsMean (±SD) ISO_MAC in the control treatment was 1.19 ± 0.15% and 1.18 ± 0.15% at 2.5 and 5 hours, respectively. The 1.0 mg kg^?1 dose of methadone reduced ISO_MAC by 48% (2.5 hours) and by 30% (5 hours), whereas the 0.5 mg kg^?1 dose caused smaller reductions in ISO_MAC (35% and 15% reductions at 2.5 and 5 hours, respectively). Both doses of methadone decreased heart rate (HR), but the 1.0 mg kg^?1 dose was associated with greater negative chronotropic actions (HR 37% lower than control) and mild metabolic acidosis at 2.5 hours. Mean arterial pressure increased in the MET1.0 treatment (13% higher than control) at 2.5 hours.Conclusions and clinical relevanceMethadone reduces ISO_MAC in a dose-related fashion and this effect is lessened over time. Although the isoflurane sparing effect of the 0.5 mg kg^?1 dose of methadone was smaller in comparison to the 1.0 mg kg^?1 dose, the lower dose is recommended for clinical use because it results in less evidence of cardiovascular impairment.