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.     

Efficacy of five selected acaricides against Tetranychus urticae (Acari: Tetranychidae) and their side effects on relevant natural enemies occurring in citrus orchards

BACKGROUND: Three groups of natural enemies are fundamental in citrus IPM in Spain: coccinellid and phytoseiid predators and hymenopteran parasitoids. Tetranychus urticae Koch is an important pest affecting citrus, for which biological control has not yet been achieved; therefore, acaricides are commonly used to control it. The goal of this study was to measure the efficacy of different acaricides on this mite and their side effects on three natural enemies relevant for citrus (Cryptolaemus montrouzieri Mulsant, Neoseiulus californicus McGregor and Aphidius colemani Viereck).RESULTS: Some products proved highly effective against T. urticae and harmless to A. colemani (mineral oil, tebufenpyrad, clofentezine and fenazaquin). However, almost all products tested were slightly harmful for both the predators considered. Fenazaquin was even moderately harmful for N. californicus. CONCLUSION: Further studies, like that presented here, are necessary to gain a better understanding of integrating biological and chemical controls. When considering both efficacy and side effects on beneficial arthropods, the best options would seem to be mineral oil, tebufenpyrad and clofentezine. However, it is urgent to complete testing of the side effects of the acaricides used in citrus. This question is crucial if the fact that two recently introduced Tetranychidae are being controlled in citrus by chemical means exclusively is considered.     

Quarantining the Sahara desert: growth and water-use efficiency of Aleppo pine in the Algerian Green Barrier

The Algerian Green Barrier, mainly composed of native and artificial Aleppo pine forests, spreads along the pre-Saharan steppes and is threatened by anthropogenic and natural disturbances, including climate change. We hypothesized that the ecophysiological functioning of this conifer has been substantially modified in reaction to recent warming and drought much beyond the expected effect of CO₂ fertilization. Our aim was to characterize the long-term performance (1925–2013) of native Aleppo pines thriving at their southernmost distribution. We used tree-ring width (TRW) and carbon isotope discrimination (Δ¹³C) to characterize basal area increment (BAI) and intrinsic water-use efficiency (WUEi) at three sites. BAI remained stable or slightly increased over time, with mean values ranging between 4.0 and 6.3 cm² year^?1. Conversely, site-Δ¹³C decreased from ?0.022 to ?0.014‰ year^?1 along time, which translated into WUEi increases of ca. 39%. This strong physiological reaction indicated that pines were responding simultaneously to rising CO₂ and drier conditions, inducing a progressively tighter stomatal control of water losses. However, WUEi increments were essentially unrelated to BAI and did not affect carbon reserves, which suggests a high resilience to climate change. This finding could be due to shifts in growing season towards earlier months in winter–spring, as suggested by temporal changes in climate factors underlying Δ¹³C and TRW. Our study highlights the substantial plasticity of Aleppo pine, but this species is unlikely to follow a similar pace of ecophysiological adjustments according to unprecedented low Δ¹³C records and lack of WUEi stimulation observed from 2000 onwards.     

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.     

Tree performance and fruit yield and quality of ‘Okitsu’ Satsuma mandarin grafted on 12 rootstocks

The citriculture in Brazil, as well as in other important regions in the world, is based on very few mandarin cultivars. This fact leads to a short harvest period and higher prices for off-season fruit. The ‘Okitsu’ Satsuma (Citrus unshiu Marc.) is among the earliest ripening mandarin cultivars, and it is considered to be tolerant to citrus canker (Xanthomonas citri subsp. citri Schaad et al.) and to citrus variegated chlorosis (Xylella fastidiosa Wells et al.). Despite having regular fruit quality under hot climate conditions, the early fruit maturation and absence of seeds of ‘Okitsu’ fruits are well suited for the local market in the summer (December through March), when the availability of citrus fruits for fresh consumption is limited. Yet, only a few studies have been conducted in Brazil on rootstocks for ‘Okitsu’. Consequently, a field trial was carried out in Bebedouro, São Paulo State, to evaluate the horticultural performance of ‘Okitsu’ Satsuma mandarin budded onto 12 rootstocks: the citrandarin ‘Changsha’ mandarin (Citrus reticulata Blanco) × Poncirus trifoliata ‘English Small’; the hybrid Rangpur lime (Citrus limonia Osbeck) × ‘Swingle’ citrumelo (P. trifoliata (L.) Raf. × Citrus paradisi Macfad.); the trifoliates (P. trifoliata (L.) Raf.) ‘Rubidoux’, ‘FCAV’ and ‘Flying Dragon’ (P. trifoliata var. monstrosa); the mandarins ‘Sun Chu Sha Kat’ (C. reticulata Blanco) and ‘Sunki’ (Citrus sunki (Hayata) Hort. ex. Tanaka); the Rangpur limes (C. limonia Osbeck) ‘Cravo Limeira’ and ‘Cravo FCAV’; ‘Carrizo’ citrange (Citrus sinensis × P. trifoliata), ‘Swingle’ citrumelo (P. trifoliata × C. paradisi), and ‘Orlando’ tangelo (C. paradisi × Citrus tangerina cv. ‘Dancy’). The experimental grove was planted in 2001, using a 6 m × 3 m spacing, in a randomized block design. No supplementary irrigation was applied. Fruit yield, canopy volume, and fruit quality were assessed for each rootstock. A cluster multivariate analysis identified three different rootstock pairs with similar effects on plant growth, yield and fruit quality of ‘Okitsu’ mandarin. The ‘Flying Dragon’ trifoliate had a unique effect over the ‘Okitsu’ trees performance, inducing lower canopy volume and higher yield efficiency and fruit quality, and might be suitable for high-density plantings. The ‘Cravo Limeira’ and ‘Cravo FCAV’ Rangpur limes induced early-ripening of fruits, with low fruit quality. ‘Sun Chu Sha Kat’ and ‘Sunki’ mandarins and the ‘Orlando’ tangelo conferred lower yield efficiency and less content of soluble solids for the latter rootstock.     

Nature-inspired soluble iron-rich humic compounds: new look at the structure and properties

Purpose

Humic substances (HS) being natural polyelectrolyte macromolecules with complex and disordered molecular structures are a key component of the terrestrial ecosystem. They have remarkable influence on environmental behavior of iron, the essential nutrient for plants. They might be considered as environmental friendly iron deficiency correctors free of synthetic iron (III) chelates disadvantages. The main goal of this study was to obtain water-soluble iron-rich humic compounds (IRHCs) and to evaluate their efficiency as chlorosis correctors.

Materials and methods

A facile preparation technique of IRHCs based on low-cost and available parent material was developed. The iron-containing precursor (ferrous sulfate) was added dropwisely into alkaline potassium humate solution under vigorous stirring and pH-control. A detailed characterization both of organic and inorganic parts of the compounds was provided, the iron species identification was carried out jointly by EXAFS and Mössbauer spectroscopy. Bioassay experiments were performed using cucumber Cucumis sativus L. as target plants. Plants were grown in modified Hoagland nutrient solution, prepared on deionized water and containing iron in the form of IRHCs. Total iron content in dry plants measured by spectrophotometry after oxidative digestion and the chlorophyll a content determined after acetone extraction from fresh plant were used as parameters illustrating plants functional status under iron deficiency condition.

Results and discussion

The high solubility (up to130 g/l) and iron content (about 11 wt%) of the IRHCs obtained allow considering them to be perspective for practical applications. A set of analytical methods has shown that the main iron species in IRHCs are finely dispersed iron (III) oxide and hydroxide nanoparticles. An application of the precursor solution acidification allows to obtain compounds containing a significant part of total iron (up to 30 %) in the form of partly disordered iron (II–III) hydroxysulphate green rust GR(SO₄ ^2?). Bioavailability of iron from IRHCs was demonstrated using bioassay in cucumber plants grown up on hydroponics under iron deficiency conditions.

Conclusions

The application of iron oxides chemistry for humic substance containing solution was proved to be an effective approach to synthesis of IRHCs. Using bioassay on cucumber plants C. sativus L. under iron deficiency conditions, the efficiency of compounds obtained as chlorosis correctors was demonstrated. Application of water-soluble IRHCs led to significant increase of chlorophyll a content (up to 415 % of the blank) and iron content in plants (up to 364 % of the blank) grown up on hydroponics.     

Virulence genes and plasmid profiles in Rhodococcus equi isolates from domestic pigs and wild boars (Sus scrofa) in Brazil

The virulence genes and plasmid profiles of 23 Rhodococcus equi isolates from 258 lymph nodes from domestic pigs (129 nodes with lesions and 129 without lesions) and 120 lymph nodes from slaughtered wild boars (60 nodes with lesions and 60 without) were characterized. R. equi was obtained from 19 lymph nodes of domestic pigs, 17 with, and two without lesions, and from four lymph nodes with lesions, from wild boars. The 23 isolates were tested for the presence of vapA and vapB genes, responsible for the 15–17 and 20 kDa virulence-associated proteins, respectively, by PCR in order to characterize as virulent (VapA), intermediately virulent (VapB) and avirulent. Plasmid DNAs were isolated and analyzed by digestion with restriction endonucleases to estimate size and compare their polymorphisms. Of the 19 domestic pigs strains, seven (36.8%) were avirulent and 12 (63.2%) were intermediately virulent, with the intermediately virulent isolates being plasmid types 8 (8 isolates), 10 (2 isolates), 1 (1 isolate) and 29 (1 isolate). The plasmid type of four strains isolated from wild boars was also intermediately virulent type 8. None of the domestic pigs and wild boar isolates showed the vapA gene. These findings demonstrate a high occurrence of plasmid type 8 in isolates from pigs and wild boars, and the similarity of plasmid types in the domestic pigs, wild boars and human isolates in Brazil.