Occurrence of Neospora caninum in dogs and its correlation with visceral leishmaniasis in the urban area of Campo Grande, Mato Grosso do Sul, Brazil

Neospora caninum is an obligate intracellular protozoan that can infect domestic and wild canids, as well as ruminants and equines, and is described as causing neuromuscular alteration and death in dogs. Visceral leishmaniasis (VL) is an infectious disease that affects both humans and animals, being caused by protozoan parasites of the genus Leishmania, of which Leishmania (Leishmania) chagasi is found in Brazil -- transmitted by sand flies, such as Lutzomyia longipalpis, in most of the American continent. The immunosuppression caused by VL can promote the occurrence of co-infections with other agents. In order to determine the frequency of N. caninum and its relationship to VL in Campo Grande, MS, Brazil, 345 blood sera were collected from dogs. The sera were submitted to an indirect fluorescent antibody test (IFAT) for detection of anti-N. caninum antibodies and VL antibodies. N. caninum was found in 26.5% of VL-negative dogs and in 29% of VL-positive ones. Among males, it was found in 30.7% of VL-negative animals and in 30.4% of VL-positive ones; among females, in 21% of VL-negative animals and in 27.7% of VL-positive ones. Among juvenile dogs (under 1 year), N. caninum was detected in 10.5% of VL-negative animals and in 11.2% of VL-positive ones. For adult dogs (1 year and older) the results were 31.4% for VL-negative animals and 28.8% of VL-positive ones. The study revealed a statistically significant association with age (chi(2)=9.76, P<0.05) in the N. caninum results for VL-negative animals. No significant correlation in N. caninum seroprevalence was found when VL-positive or VL-negative dogs were compared (chi(2)=0.21, P=0.64). The findings suggest that in Campo Grande N. caninum and VL co-infection is common in dogs, though VL does not appear to enhance susceptibility to N. caninum.     

Cacao,copoazu and macambo: Exploring Theobroma diversity in smallholder agroforestry systems of the Peruvian Amazon

Over the past decades, the general trend towards shade reduction and intensification of cacao management has led to biodiversity losses. In the Peruvian Amazon, the regional government is heavily promoting crop conversion to shift from regionally marketed foods towards cacao (Theobroma cacao) and copoazu (Theobroma grandiflorum). While this shift is already visibly impacting the farming landscape and the lives of many smallholder farmers, little is known about the reasons that drive farmers to choose certain types of Theobroma species or cacao varieties over others. In this paper, we addressed how cacao farmers perceive and manage specific and varietal Theobroma diversity. We interviewed cacao farmers (n?=?20) during a seed-exchange fair and adapted a version of the four-square analysis to explore which Theobromas are currently adopted by farmers and why. The native cacao variety (cacao chuncho) was the one cultivated by most farmers, followed by the more industrial clonal varieties. The source of seeds and seedlings for the most cultivated varieties was a mix of donations by public institutions (for clonal varieties) and informal exchange among farmers (mainly for native and criollo varieties, and species i.e. macambo (Theobroma bicolor) and cacauillo (Theobroma speciosum)). The cacao varieties incentivized by public institutions were the least desired for future investment. The motivations for farmers to plant Theobromas was mainly based on the perceived current and potential market, but their desire to invest in a given species or variety was also based on pest resistance, traditional use, farm diversification and taste. Copoazu and CCN cacao varieties are produced by many households in large areas, while macambo, cacauillo and the cacao varieties TSH and porcelana are produced by few households in small areas. Our study suggests that agroforestry systems that include alternative Theobroma species are multi-strata and more diverse than cacao-based systems. It highlights the risks of agrobiodiversity loss associated with the promotion of industrial cacao varieties, and the importance of seed and seedling access for the development of diverse farming systems. We recommend the expansion of local seed-sharing networks and the extension of public cacao-donation campaigns to neglected and underutilized Theobromas with socio-economic and environmental benefits, in order to make the local farming systems more diverse and resilient.

     

Development of an indirect ELISA-NcSRS2 for detection of Neospora caninum antibodies in cattle

Neosporosis is of alarming economic concern in the cattle industry. The effectiveness of diagnostic tests for detecting specific antibodies against Neospora caninum is hampered by potential cross-reaction with other coccidia. Use of a single specific antigen might improve test specificity. An indirect enzyme-linked immunosorbent assay (ELISA) was developed using the truncated protein NcSRS2 expressed in Escherichia coli. The ELISA results were compared with those of the indirect fluorescence antibody test (IFAT). Receiver Operating Characteristic (ROC) and Tests in the Absence of a Gold Standard (TAGS) analysis revealed an assay having 96% specificity and 95% sensitivity when applied to 145 positive and 352 negative sera from two distinct cattle populations. Using OD ≤ 0.095 as the cut-off point, the assay's negative and positive predictive values ranged from 98.8% to 50.8% and from 58.8% to 99.1%, respectively, depending on neosporosis prevalence in a given area. The novel ELISA-NcSRS2 format described in the present report constitutes a specific and sensitive method for detecting N. caninum in cattle.     

Innovative light management to improve production sustainability,overall quality,and the phenolics composition of nectarine (Prunus persica cv. Stark Red Gold)

Summary

This research was conducted under the framework of the ISAFRUIT Project and aimed to investigate the effects of different light micro-environments on the final overall quality of nectarine fruit production. Experiments were conducted in a commercial orchard of the nectarine (Prunus persica) ‘Stark Red Gold’ during 2006 and 2007. Reflective mulches were laid down in the inter-row spaces in mid-May. Those environmental conditions affected by mulching such as temperature and reflected light were monitored until fruit harvest. Fruit production per tree was enhanced by mulch-treatment in both years, but the differences were statistically significant only in 2006, when the average fruit weight was also enhanced. Nectarines became more ripe in the 2006 season, whereas no differences in the main fruit quality indices were detected in 2007.The concentrations of phenolic compounds in ripe nectarines were positively enhanced in both years. The increase in concentration of overall phenolic compounds (in mg 100 g^?1 fresh weight) was calculated to be approx. 60% in 2006 and 2007, indicating an interesting improvement of the nutraceutical and anti-oxidant potential of nectarines. Experiments were also conducted using UV plus white light irradiation under controlled conditions. The accumulation of phenolic compounds, specifically anthocyanin concentrations, in nectarines previously screened using paper bags were determined at different times after irradiation. The results clearly indicated an inducing effect of UV plus white light irradiation on the synthesis and accumulation of anthocyanins in fruit skin. The consequences for the colour and health potential of nectarines are discussed.     

Synthetic pyrazole derivatives as growth inhibitors of some phytopathogenic fungi

The present study was carried out to investigate the antifungal activity of pyrazole/isoxazole-3-carboxamido-4-carboxylic acids, 4-oxo-5-substituted pyrazolo[3,4-d]pyrimidine-6-thiones, and N-alkyl/aryl-N'-(4-carbethoxy-3-pyrazolyl)thioureas against Pythium ultimum, Botrytis cinerea, and Magnaporthe grisea. The results on growth inhibition showed differences in the sensitivity of the three fungi to the tested substances, and in general P. ultimum was shown to be the most sensitive. On all phytopathogens the best results within the pyrazole/isoxazolecarboxamide series are given by the compounds with the carboxamide and carboxylic groups in positions 3 and 4; the presence of these groups seems to be critical for biological activity in this series of compounds. Among the pyrazolopyrimidines the derivative supplied with the benzylic group was the most active on the three fungi and in particular against P. ultimum. Several compounds belonging to the thiourea series are able to inhibit selectively M. grisea at 50 and 10 microg mL(-1), doses at which the reference commercial compound tricyclazole had low or no effect.     

Wheat Nitrogen Fertilization Under no Till on the Low Altitude Brazilian Cerrado

This study evaluates the effects of different nitrogen (N) rates (0, 50, 100, 150, and 200 kg ha^?1) and sources (ammonium sulfate, urea, and ammonium sulfonitrate with a nitrification inhibitor, Entec®) applied either as a single application during sowing (next to the planting rows) or as side dressing (stage 3.2 on Zadok scale), on production components and grain yield of irrigated wheat grown under no-till in a low-elevation Brazilian cerrado and Dark Red Dystrophic Latosol with clayey-texture. It was adopted a randomized block design (5 × 3 × 2) in factorial scheme with four replications. The N sources provided similar grain yields and chlorophyll contents. Increased nitrogen rates increased grain yield up to the 120 kg N ha^?1, regardless of application time and N source. Nitrogen fertilization as side dressing can be recommended based on leaf chlorophyll readings. Early application of all nitrogen fertilizers at sowing time was viable.     

Determinants of high pesticide exposure events in the agricultural health cohort study from enrollment (1993-1997) through phase II (1999-2003)

We conducted an analysis of the determinants of high pesticide exposure events (HPEEs), which are defined as self-reported incidents of high exposure to pesticides, fertilizers, or other chemicals in the Agricultural Health Study, a cohort of private applicators and their spouses residing in North Carolina or Iowa, and commercial applicators residing in Iowa. We examined the risk of HPEEs occurring between enrollment (phase 1: 1993-1997) and follow-up (phase II: 1999-2003) among participants who completed the phase II questionnaire (n=43,149) by calculating hazard rate ratios and 95% confidence intervals using Cox proportional-hazard regression. During the followup period, 1,582 HPEEs were reported (3.8%). HPEE risk was significantly higher among Iowa residents, younger participants, those with a hearing deficit, a risk-taking personality, and an HPEE prior to enrollment. Among private applicators (n=30,102), larger farm size, higher frequency and duration of pesticide use, spraying pesticides with open cab windows, using a tractor cab without a charcoal filter, repairing spray equipment, wearing work clothing more than two days without changing, not removing work boots before entering the home, and storing pesticides in the home were associated with significantly higher HPEE risk. Among commercial applicators (n=2326), higher frequency of pesticide use was associated with a significantly higher HPEE risk. Among spouses (n=10,721), higher frequency of pesticide use, using an application vehicle with a cab, and storing pesticides in the home were associated with a significantly higher HPEE risk. Our findings indicate that HPEEs were associated with several modifiable pesticide handling procedures that can be targeted in safety training and education.     

Positive regulation of Itk PH domain function by soluble IP4

Pleckstrin homology (PH) domain-mediated protein recruitment to cellular membranes is of paramount importance for signal transduction. The recruitment of many PH domains is controlled through production and turnover of their membrane ligand, phosphatidylinositol 3,4,5-trisphosphate (PIP3). We show that phosphorylation of the second messenger inositol 1,4,5-trisphosphate (IP3) into inositol 1,3,4,5-tetrakisphosphate (IP4) establishes another mode of PH domain regulation through a soluble ligand. At physiological concentrations, IP4 promoted PH domain binding to PIP3. In primary mouse CD4+CD8+ thymocytes, this was required for full activation of the protein tyrosine kinase Itk after T cell receptor engagement. Our data suggest that IP4 establishes a feedback loop of phospholipase C-gamma1 activation through Itk that is essential for T cell development.     

Synthesis and antifungal action of new tricyclazole analogues

Melanins are very important pigments for the survival and longevity of fungi, so their biosynthesis inhibition is a new biochemical target aiming at the discovery of selective fungicides. In this work is described the synthesis of new pyrazolo-thiazolo-triazole compounds, analogues of tricyclazole (a commercial antifungal product that acts by inhibiting melanin synthesis), and their biological activity was studied on some dermatophytes and phytopathogens. The compounds poorly inhibited the growth and pigmentation of fungi tested and were less efficient than tricyclazole. Electron microscopy on Botrytis cinerea showed that treatment with the most active compound caused abnormally thickened and stratified walls in fungi, whose ultrastructure was, in contrast, generally normal. The fungus treated with tricyclazole, on the other hand, appeared to be drastically altered, so as to become completely disorganized. These results suggest that the new azole compounds employ an action mechanism similar to that of other azoles, but dissimilar to that of tricyclazole.