Acute lethal and behavioral sublethal responses of two stored-product psocids to surface insecticides

BACKGROUND: The psocids Liposcelis bostrychophila Badonnel and L. entomophila (Enderlein) (Liposcelididae) are emerging pests of stored products. Although their behavior, particularly their high mobility, may contribute to the reported relatively low efficacy of insecticides against them, studies to investigate this have not been conducted. The present study aimed to assess the label rate efficacy of three commercial insecticides (β‐cyfluthrin, chlorfenapyr and pyrethrins) applied on concrete surfaces against L. bostrychophila and L. entomophila, and also their sublethal effect on the mobility of these species. RESULTS: The synthetic insecticides β‐cyfluthrin and chlorfenapyr showed high short‐term efficacy (LT₉₅≤15 h) against both psocid species, unlike the natural pyrethrins (LT₉₅ ≥ 4 days). Liposcelis bostrychophila was slightly more tolerant (≥1.2×) than L. entomophila to all three insecticides. Behavioral assays with fully sprayed and half‐sprayed concrete arenas indicated that the insecticides reduced the mobility of both species. Pyrethrins seem to elicit weak repellence in L. bostrychophila. CONCLUSION: β‐Cyfluthrin and chlorfenapyr were effective against both psocid species, but not pyrethrins. The mobility of both species does not seem to play a major role in the differential selectivity observed, but the lower mobility of L. bostrychophila may be a contributing factor to its higher insecticide tolerance. Published in 2008 by John Wiley & Sons, Ltd.     

Metabolic signature of spent culture media shows lipid metabolism as a determinant of pregnancy outcomes

In the present study, we investigated the spent culture media of in vitro produced (IVP) bovine embryos which did (group Pregnant) or did not (group Non-pregnant) establish pregnancy after transfer. For that purpose, IVP embryos on D5 were transferred to individual droplets for the last 48 h of culture. Embryos at the blastocyst stage were then transferred to synchronized recipients, while respective culture media drops were collected and evaluated individually. The list of metabolites present in spent culture media was obtained by electrospray ionization mass spectrometry (ESI-MS) and analysed with Metaboanalyst® to characterize the metabolic profile of each group. The spectrometric analysis showed that pathways related to lipid metabolism, particularly fatty acids degradation via beta-oxidation, were more present in the Pregnant group whereas no significant pathway was identified in the group Non-pregnant. By using this method, we were able to identify a metabolic signature in culture media that allows for a better comprehension of preferential metabolic routes taken by the most viable embryos. These findings offer great insights into the biochemistry of embryo development and reveal a potential target for the development of better-quality IVP systems, as well as tools to identify bovine embryos with greater chances to establish and maintain pregnancy.     

Midgut cysteine-proteinase activity in the velvetbean caterpillar (<Emphasis Type="Italic">Anticarsia gemmatalis</Emphasis> (Hübner))

Proteinase inhibitors are currently targeted as potential insect control agents, but adaptation to proteinase inhibitors is a recognized limitation to such approach requiring the understanding of how phytophagous species can cope with such compounds. The velvetbean caterpillar (Anticarsia gemmatalis) is a key pest of soybean and well-adapted to its host proteinase inhibitors, which is rich in serine-proteinase inhibitors, particularly trypsin-like proteinase inhibitors. As the expression of cysteine proteinases in the midgut of the velvetbean caterpillar is a potential adaptation to circumvent its host defense, we assessed and characterized the digestive cysteine-proteinase activity from velvetbean caterpillars. Significant soluble and membrane-bound proteolytic activity was obtained and was consistent with those of cysteine proteinases based on the substrate and inhibitors used for their characterization. The K _m and V _max values obtained were 2.35 ± 0.50 mM and 40.89 ± 6.68 nmol min⁻¹ mg⁻¹ for the soluble proteinases and 0.33 ± 0.03 mM and 24.54 ± 0.67 nmol min⁻¹ mg⁻¹ for the membrane-bound proteinases, range of values also consistent with cysteine proteinases. Therefore, the proteolytic activity observed in the velvetbean caterpillar midgut is consistent with that of cysteine proteinases providing preliminary support for the contention of their potential involvement mitigating the negative effects of serine-protease inhibitors in this species.     

Qualitative and quantitative polymerase chain reaction (PCR) for detection of Leishmania in spleen samples from naturally infected dogs

Because infected dogs are widely considered to be the main domestic reservoir for Leishmania infantum (syn Leishmania chagasi) parasites in Brazil, the diagnosis of canine visceral leishmaniasis (CVL) must be made both accurately and promptly. The present study attempted to standardize a conventional polymerase chain reaction (cPCR) protocol for the detection of L. infantum DNA in canine spleen samples. Quantitative PCR (qPCR) technique was used to confirm the presence of Leishmania DNA in the canine spleen fragments. A comparison was made between the efficacies of these molecular diagnostic techniques and conventional parasitological and serological methods. cPCR protocols for spleen samples were standardized using primers that amplify a 145 bp fragment, located at the parasite kinetoplast minicircle. The genus specificity of the cPCR protocol was assessed by its inability to amplify the DNA of other common canine pathogens, such as Ehrlichia canis, Babesia canis, Toxoplasma gondii and Trypanosoma cruzi. cPCR protocol sensitivity was tested by assessing the reaction detection limit, determined to be 10 fg of L. infantum reference strain DNA, which corresponds to a range of 0.03-0.1 parasites per fragment. Standardized cPCR protocol was used to detect the presence of Leishmania in 45 dog spleen samples. Our results showed that 40% of the spleen fragment cultures were positive for Leishmania parasites, 58% of the dog serum samples tested positive using ELISA, and parasite DNA was detected in 44% using qPCR, while 47% of the spleen samples using cPCR. Diagnostic methods performance was assessed and revealed a better degree of ascertainment for cPCR when compared to other diagnostic methods. The sensitivity of ELISA was 83.3%, qPCR was 83.3%, and cPCR was 88.9%; PPV for ELISA was 57.7%, qPCR was 75% and cPCR was 76.2%; the Kappa coefficients were found to be 0.40 (fair) for ELISA, 0.64 (substantial) for qPCR and 0.68 (substantial) for cPCR. In both oligosymptomatic and polysymptomatic dogs, cPCR revealed the better performance analysis when compared to other diagnostic methods. The findings presented herein establish cPCR as the most indicated test to detect Leishmania when compared to the other two diagnostic methods evaluated. Despite the fact that the qPCR protocol provides a highly accurate quantification of parasites when targeting the SSU rRNA gene, this technique does not significantly improve the diagnosis of CVL when compared with the performance of the cPCR protocol, which focused on the kinetoplast minicircle.     

Application of a multiplex PCR assay for Campylobacter fetus detection and subspecies differentiation in uncultured samples of aborted bovine fetuses

Campylobacter (C.) fetus (epsilonproteobacteria) is an important veterinary pathogen. This species is currently divided into C. fetus subspecies (subsp.) fetus (Cff) and C. fetus subsp. venerealis (Cfv). Cfv is the causative agent of bovine genital Campylobacteriosis, an infectious disease that leads to severe reproductive problems in cattle worldwide. Cff is a more general pathogen that causes reproductive problems mainly in sheep although cattle can also be affected. Here we describe a multiplex PCR method to detect C. fetus and differentiate between subspecies in a single step. The assay was standardized using cultured strains and successfully used to analyze the abomasal liquid of aborted bovine fetuses without any pre-enrichment step. Results of our assay were completely consistent with those of traditional bacteriological diagnostic methods. Furthermore, the multiplex PCR technique we developed may be easily adopted by any molecular diagnostic laboratory as a complementary tool for detecting C. fetus subspecies and obtaining epidemiological information about abortion events in cattle.     

Environmental Behaviour of Metolachlor and Diuron in a Tropical Soil in the Central Region of Brazil

The environmental behaviour of metolachlor and diuron was studied in the Central-western region of Brazil, by means of a field study where six experimental plots were installed. The soil was classified as a Latosol, and the soil horizons were characterized. Sorption of metolachlor and diuron was evaluated in laboratory batch experiments. Metolachlor and diuron were applied to the experimental plots on uncultivated soil in October 2003. From this date to March 2004, the following processes were studied: leaching, runoff and dissipation in top soil. K _oc of metolachlor varied from 179 to 264 mL g^?1 in the soil horizons. K _oc of diuron in the Ap horizon was 917 mL g^?1, decreasing significantly in the deeper horizons. Field dissipation half-lives of metolachlor and diuron were 18 and 15 days, respectively. In percolated water, metolachlor was detected in concentrations ranging from 0.02 to 2.84 μg L^?1. In runoff water and sediment, metolachlor was detected in decreasing concentrations throughout the period of study. Losses of 0.02% and 0.54% of the applied amount by leaching and runoff, respectively, were observed confirming the high mobility of this herbicide in the environment. In percolated water, diuron was detected with low frequency but in relatively high concentrations (up to 6.29 μg L^?1). In runoff water and soil, diuron was detected in decreasing concentrations until 70 days after application, totalizing 13.9% during the whole sampling period. These results show the importance of practices to reduce runoff avoiding surface water contamination by these pesticides, particularly diuron.