Specificity of the carbon immunoassay (CIA) test for the diagnosis of Toxoplasma infections.

A rapid and low cost procedure, the carbon immunoassay (CIA) test, was evaluated for the diagnosis of Toxoplasma gondii infections. Using a closely related parasite (Besnoitia jellisoni) as antigen, and homologous or heterologous immune sera, it was demonstrated by light and electron microscopy that CIA is a very reliable and specific test. As it is neither expensive nor time-consuming, it can be recommended for general and routine laboratory use.     

Molecular cloning,purification and immunogenicity of recombinant Brucella abortus 544 malate dehydrogenase protein

The Brucella mdh gene was successfully cloned and expressed in E. coli. The purified recombinant malate dehydrogenase protein (rMDH) was reactive to Brucella-positive bovine serum in the early stage, but not reactive in the middle or late stage, and was reactive to Brucella-positive mouse serum in the late stage, but not in the early or middle stage of infection. In addition, rMDH did not react with Brucella-negative bovine or mouse sera. These results suggest that rMDH has the potential for use as a specific antigen in serological diagnosis for early detection of bovine brucellosis.     

Diversity of insecticide resistance mechanisms and spectrum in European populations of the codling moth, Cydia pomonella

Only a few of the registered insecticides against Cydia pomonella L. are still effective in areas where insecticide resistance has emerged in this pest. Resistance mechanisms are multiple, and their lone or cumulative effects in a single population are not completely understood. A detailed estimation of resistance spectrum is still required to define the suitable insecticides to use against a given population. The efficacy of ten insecticides was therefore investigated together with the resistance mechanisms expressed in four laboratory strains and 47 field populations of C. pomonella from five countries. Bioassays were performed using topical applications of diagnostic concentrations on diapausing larvae, and resistance mechanisms were analysed on adults emerging from control insects. All populations exhibited a reduced susceptibility to at least one insecticide when compared with the susceptible laboratory strain. Cross-resistances were observed between azinphos-methyl or phosalone and more recent compounds such as spinosad and thiacloprid. Resistances to azinphos-methyl, diflubenzuron, spinosad, tebufenozide and thiacloprid were significantly correlated with mixed-function oxidase activity, while increased glutathione-S-transferase and reduced non-specific esterase activities were correlated with resistance to azinphos-methyl and emamectin, respectively. Conversely, resistances to azinphos-methyl, tebufenozide and thiacloprid were negatively correlated with increased esterase activity. None of the observed mechanisms explained the loss of susceptibility of populations to chlorpyrifos-ethyl, and no significant correlation was detected between resistance to deltamethrin and the presence of the kdr mutation. The suitability of such non-target instars to monitor insecticide resistance in field populations is discussed.     

Resistance monitoring in codling moth: a need for standardization

BACKGROUND: A wide range of methods has been used for monitoring resistance in Cydia pomonella L. However, the results from specific tests are not always concordant and they generate doubts over the extent to which the data represent the field situation. Therefore, the variability in the expression of resistance mechanisms and in resistance ratios between developmental stages and their reliability in a field situation were studied. RESULTS: Neonate larvae showed the highest insecticide resistance, except to spinosad. Resistance ratios were from 2 to 9.5, depending on the insecticide and the strain tested, when compared with the susceptible strain. Spinosad exhibited the highest resistance ratio for diapausing larvae (4.4, 12.2 and 4.4 for Raz, Rdfb and RΔ strains, respectively). Enzymatic ratios were higher in adults than in neonates or diapausing larvae, and the highest enzymatic ratios were observed for mixed‐function oxidases (7.3–36.7 for adults of resistant strains). CONCLUSIONS: The sensitivity of different instars depends on the insecticides used, and there is a differential expression of resistance mechanisms between stages. For the population considered in this study, tests on neonates provided the most consistent results to the field situation, but adult response to chlorpyrifos‐ethyl and azinphos‐methyl in the laboratory was also consistent with the field results. Copyright © 2008 Society of Chemical Industry     

Biocultural Diversity of Sarangani Province,Philippines: An Ethno-Ecological Analysis

This paper discussed effects of lowland-associated influences on upland ecology, food security and biocultural diversity in the Sarangani farming communities of the Philippines. In the uplands of Sarangani Province, the conservation of traditional rice varieties, the centrality of rice in tribal life, and the continued observance of planting rituals attest to its cultural significance and convey a common desire for cultural preservation and community solidarity. Economic and socio-political pressures had transformed tribal communities, although vestiges of traditional farming systems are still being practiced in remote sitios(villages). Changing land use patterns had also resulted in shrinking farm sizes and consequently in food insecurity in the Sarangani uplands. Extractive industries(i.e. logging, mining and charcoal making) and swidden farming were observed to cause widespread environmental degradation, while modern agriculture had undermined the capacity of indigenous peoples to survive because of their complete dependence on lands and resources. With the reality that cultural and biological diversities are inextricably linked, trans-disciplinary strategies coupling indigenous knowledge systems with scientific knowledge should, therefore, be instituted to save the Sarangani upland ecosystem, the indigenous peoples and their tribal resources.     

Susceptibility of the Mexican bean beetle <Emphasis Type="Italic">Epilachna varivestis</Emphasis> Mulsant (Coleoptera: Coccinellidae) to endemic isolates of <Emphasis Type="Italic">Beauveria bassiana</Emphasis> (Balsamo) Vuillemin (Hypocreales: Clavicipitaceae)

Entomopathogenic fungi represent excellent candidates for biological insecticides. Among the many entomopathogenic fungi, Beauveria bassiana (Balsamo) Vuillemin (Hypocreales: Clavicipitaceae) is one of the most promising species. Here, we report the results of infection studies performed using four B. bassiana native isolates (Bb37, Bb38, Bb40, and Bb45) compared with the commercial product Mycotrol^® (strain GHA) on adult Mexican bean beetle Epilachna varivestis Mulsant (Coleoptera: Coccinellidae), one of the most important pests of bean production in Mexico. First, single concentration (1 × 10⁸ conidia mL^?1) assays were carried out using all isolates applied using an immersion method. Examinations at 10 days after inoculation indicated that isolates Bb37, Bb38, and Bb40 significantly reduced adult survival, causing 60–75% mortality compared to the commercial strain GHA (33%) and control (29%). Second, using two selected isolates (Bb37 and Bb40), the mean lethal concentration values (LC₅₀) were estimated for third instar larvae and adult E. varivestis. The LC₅₀ values for Bb37 and Bb40 isolates decreased 5.26-fold and 3.19-fold, respectively, in third instars compared with adults. However, such difference for Bb40 isolate was not significant. Finally, an experiment, to compare the median survival time (MST) values between third instar and adult E. varivestis, was conducted using Bb37 and Bb40 at 1 × 10⁹ conidia mL^?1. MST values were significantly reduced against larvae (87 and 100 h) compared with adults (130 and 134 h) by Bb37 and Bb40, respectively. We conclude that native isolates of B. bassiana represent an important alternative for the control of E. varivestis. However, to demonstrate its effectiveness under field conditions, detailed studies are needed.     

Quantification of airborne spores of <Emphasis Type="Italic">Monilinia fructicola</Emphasis> in stone fruit orchards of California using real-time PCR

The fungal pathogen Monilinia fructicola causes blossom blight and fruit brown rot of stone fruits in California. In this study, spore densities in the air were monitored in six orchard/year combinations with Burkard spore traps. A real-time PCR assay was developed to efficiently quantify the dynamics of spore density in these orchards during the growing season. Different patterns of dynamics of spore density were observed in these orchards. A linear relationship between numbers of spores counted with a compound microscope and those determined with the real-time PCR assay was obtained, using the same samples of spore traps. Spore density in five of six orchard/year combinations ranged from 0.0 to 0.05 spores l⁻¹, except for that in orchard 4, which showed much higher values of spore density in the air, as well as higher values and wider range of incidences of blossom infection and fruit rot than those in the other orchards. The results demonstrated a potential method to quantitatively determine spore inoculum potential in orchards by using a real-time PCR assay.     

Purification and characterization of a protease from Actinobacillus pleuropneumoniae serotype 1, an antigen common to all the serotypes.

A high molecular-mass proteolytic enzyme of Actinobacillus pleuropneumoniae serotype 1, was purified from culture supernatants (CSN) by using DEAE-cellulose and sepharose-4B-gelatin chromatography. In 10% SDS-polyacrylamide gels copolymerized with porcine gelatin, the protease showed a single band of activity of > 200 kDa. However, minor molecular-mass proteolytic bands were observed when the protease was electrophoresed in the presence of either 5% beta-mercaptoethanol, 50 mM dithiothreitol, or 0.25 M urea. Furthermore, when the > 200-kDa purified protein was passed through a sucrose gradient, several bands with proteolytic activity were found: 62, 90, 190, and 540 kDa. The proteolytic activity was increased in the presence of calcium or zinc and was not affected after being heated at 90 degrees C for 5 min. Proteolytic activities were also observed in CSN from all A. pleuropneumoniae serotypes and biotypes. The purified protease hydrolyzed porcine IgA and IgG in vitro. In addition, by immunoblot the protease was recognized by serum of naturally infected pigs with serotypes 1 and 5, and by serum of pigs experimentally infected with serotypes 1, 2, 8, or 9. Serum of a pig vaccinated with CSN of a serotype 3 strain also recognized the protease, but not sera of pigs vaccinated with a bacterin (serotype 1). Proteins from CSN of all the serotypes, which were precipitated with 70% (NH4)2SO4, were recognized by a polyclonal antibody raised against the purified protease. Taken together these results indicate that an antigenic protease is produced in vivo by all the serotypes of A. pleuropneumoniae. The results indicate that proteases could have a role in the disease and in the immune response of pigs infected with A. pleuropneumoniae.     

Bioavailability of calcium from milk-based formulas and fruit juices containing milk and cereals estimated by in vitro methods (solubility, dialyzability, and uptake and transport by caco-2 cells)

An adequate calcium intake during the first years of life is needed for normal growth and development and to prevent rickets. The bioavailability of calcium from infant foods (milk-based formulas and fruit juices containing milk and cereals, FMC), the dietary sources of calcium in these stages of life, has been estimated on the basis of simulated gastrointestinal digestion and calcium solubility and dialyzability values and on the efficiency of transport and uptake by Caco-2 cells. The ranking of samples according to calcium bioavailability depends on the use of solubility or dialyzability as criterion. On the basis of the former, the highest value corresponded to adapted formulas and the lowest to fruit juices. However, when using percentage dialysis, the highest value corresponded to fruit juices and the lowest to follow-up formulas. The highest percentages of transport efficiency and uptake by Caco-2 cells corresponded to calcium from the analyzed fruit juices, followed by toddler, follow-up, and adapted formulas.     

Do mature pine plantations resemble deciduous natural forests regarding understory plant diversity and canopy structure in historically modified landscapes?

We compared the structure of the arboreal layer and the diversity and species composition of the understory vegetation of three types of mature forest communities: oak (Quercus pyrenaica) and beech (Fagus sylvatica) forests and Scots pine (Pinus sylvestris) plantations. Our main aim was to determine whether differences in these variables existed and were due to the identity of the dominant tree species. We selected four stands or replicates per forest type located geographically close and with relatively similar conditions. We found no differences in the arboreal structure of oak and beech forests, which were characterised by great variability in tree size, while in case of plantations, this variability was lower at both the intra-stand (estimated by the coefficient of variation) and inter-stand (i.e. the four replicates harboured trees of similar sizes) scales. However, the highest variability in the canopy layer of natural forests was not consistently linked to greater understory species richness. Indeed, the lowest plant species richness was found in beech forests, while oak forests harboured the highest value at either the sampling unit (per m²) or stand scales. The greatest negative correlation between plant diversity and the environmental variables measured was found for litter depth, which was the highest in beech forests. The results obtained by the CCA indicated that the four replicates of each forest type clustered together, due to the presence of characteristic species. We concluded that pine plantations did not approach the environmental conditions of native forests, as plantations were characterised by singular understory species composition and low arboreal layer variability, compared to natural woodlands.