Captan and fenitrothion dissipation in field-treated cauliflowers and effect of household processing

Field trial studies have been performed with captan and fenitrothion on cauliflower to propose maximum residue limits and to study the dissipation of the pesticides. Residue levels have been determined at different times following good laboratory practice using gas chromatography with mass spectrometric detection. The behaviour of residue levels of these compounds after household processing has been analysed using gas chromatography with electron capture detection. Seven days after treatment, residue levels of captan could be detected, but not of fenitrothion. The half-lives of dissipation for captan and fenitrothion were calculated as 0.9 and 1.8 days respectively. Washing did not significantly affect the residual amounts of captan and fenitrothion observed in raw vegetables; however, after cooking, captan had degraded completely, whereas residue levels of fenitrothion were not modified significantly.     

Study of peripheral blood cell populations involved in the immune response of goats naturally infected with Mycobacterium caprae

Tuberculosis in goats caused by Mycobacterium bovis and Mycobacterium caprae has noteworthy sanitary and economic implications. Current diagnostic assays are based on cellular immunity and although they have demonstrated a high sensitivity, some animals remain undetected. In the present study, flow cytometry has been used to determine changes in CD4+, CD8+ and CD25+ T cell populations in peripheral blood from naturally infected goats. Proportion of lymphocytes producing PPD-specific interferon-gamma (IFN-γ) was calculated and an ELISA for detection of PPD-specific IFN-γ was performed to measure the cytokine in plasma. The infected goats showed percentages of CD4+ T cells between 27.31% and 47.23% and there were not significant differences (p=0.113) with the non-infected control goats although the mean percentage was lower in this group. Regarding CD8+ T cells, a higher percentage was observed in healthy goats compared to controls (p=0.081). The mean percentage of lymphocytes expressing CD25 without antigen stimulation (30.65±3.91) was higher in lesion and/or culture-positive animals than in the controls (21.84±1.21; p=0.053). The percentage of CD4+/IFN+ T cell population stimulated with bovine PPD was a reliable marker of infection, since the mean percentage in the infected goats was significantly higher than in the controls (p<0.05). Tuberculosis in goats caused by M. caprae induced changes in cellular populations similar to those described for M. bovis in cattle.     

Evaluation of peripheral blood lymphocyte subsets in family-owned dogs naturally infected by Ehrlichia canis

Previous research suggested that clinical manifestations, histopathological lesions, and even infection maintenance in the course of canine monocytic ehrlichiosis (CME) are directly related to the immune response developed by the host. In the present study, blood lymphocyte subsets were analyzed by multiparametric flow cytometry in 37 dogs with naturally occurring CME and 47 healthy dogs used as controls. T, T helper (Th), T cytotoxic (Tc), B, non-T, non-B lymphocytes and those that express MHC class II were characterized in every dog. Animals with CME showed higher relative values of T and Tc cells and a higher absolute number of Tc cells in peripheral blood. The percentage of Th cells and the absolute and relative values of B cells were higher in healthy animals than in CME-affected dogs. The significance of these changes on the pathogenesis of natural Ehrlichia canis infection in dogs needs further evaluation.     

Partitioning of soil water among tree species in a Brazilian Cerrado ecosystem

Source water used by woody perennials in a Brazilian savanna (Cerrado) was determined by comparing the stable hydrogen isotope composition (deltaD) of xylem sap and soil water at different depths during two consecutive dry seasons (1995 and 1996). Plant water status and rates of water use were also determined and compared with xylem water deltaD values. Overall, soil water deltaD decreased with increasing depth in the soil profile. Mean deltaD values were -35 per thousand for the upper 170 cm of soil and -55 per thousand between 230 and 400 cm depth at the end of the 1995 dry season. Soil water content increased with depth, from 18% near the surface to about 28% at 400 cm. A similar pattern of decreasing soil water deltaD with increasing depth was observed at the end of the 1996 dry season. Patterns consistent with hydraulic lift were observed in soil profiles sampled in 1995 and 1997. Concurrent analyses of xylem and soil water deltaD values indicated a distinct partitioning of water resources among 10 representative woody species (five deciduous and five evergreen). Among these species, four evergreen and one deciduous species acquired water primarily in the upper soil layers (above 200 cm), whereas three deciduous and one evergreen species tapped deep sources of soil water (below 200 cm). One deciduous species exhibited intermediate behavior. Total daily sap flow was negatively correlated with xylem sap deltaD values indicating that species with higher rates of water use during the dry season tended to rely on deeper soil water sources. Among evergreen species, minimum leaf water potentials were also negatively correlated with xylem water deltaD values, suggesting that access to more readily available water at greater depth permitted maintenance of a more favorable plant water status. No significant relationship between xylem water deltaD and plant size was observed in two evergreen species, suggesting a strong selective pressure for small plants to rapidly develop a deep root system. The degree of variation in soil water partitioning, leaf phenology and leaf longevity was consistent with the high diversity of woody species in the Cerrado.     

Digestive enzyme activity at different developmental stages of blackspot seabream, Pagellus bogaraveo (Brunnich 1768)

Blackspot seabream, Pagellus bogaraveo (Brunnich), has been identified as a potential species to diversify European aquaculture production. Although rearing aspects have been widely investigated, little information exists on the nutritional requirements for this species. The aim of this study was to build up information on the activity of digestive enzymes at certain developmental stages of blackspot seabream in order to understand the nutritional needs of larvae and post larvae. Fish larvae were reared from hatching to 55 days after hatching (dah), and the feeding plan consisted in rotifers (5–35 dah), Artemia naupli (30–35 dah) metanaupli (35–45) and Gemma microdiet (45–55 dah). At 7, 11, 21, 45 and 55 days after hatching (dah), pooled samples of fish larvae were collected for analysis of trypsin, amylase, lipase, alkaline phosphatase and leucine–alanine peptidase activity. Up to 21 dah, the whole larvae body was used for enzymatic analysis, whereas in older larvae only the dissected abdominal cavity was used. Blackspot seabream body dry weight growth was exponential, increasing from 60 μg at 5 dah to 30±9.7 mg at 55 dah. Amylase specific activity decreased significantly during development, exhibiting at 11 dah (0.6 U mg^?1 protein) an average value 2.7 times lower than at 7 dah, and remaining stable between 45 and 55 dah (0.7 U mg protein^?1). Trypsin specific activity remained constant until 21 dah (between 38 and 44 mU mg protein^?1), which could be related to the larvae feeding regime. At later stages of development, lipase‐specific activity exhibited a significant increase (P<0.05), being three times higher at 55 dah (8 U mg protein^?1) than at 45 dah. The total activity of the studied digestive enzymes increased significantly during larval development (until 21 dah), whereas afterwards only lipase and leucine–alanine peptidase increased significantly between 45 and 55 dah. The pattern of digestive enzymes activity was related to organogenesis and the type of food used at different developmental stages.     

Peptides from Chia Present Antibacterial Activity and Inhibit Cholesterol Synthesis

In previous studies, it has not been reported that protein isolated from chia interferes favorably with antibacterial activity, and reduces cholesterol synthesis. The objective of this study was to determine whether commonly used commercial microbial proteases can be utilized to generate chia protein-based antibacterial and hypocholesterolemic hydrolysates/peptides, considering the effects of protein extraction method. Alcalase, Flavourzyme and sequential Alcalase-Flavourzyme were used to produce hydrolysates from chia protein (CF), protein-rich fraction (PRF) and chia protein concentrates (CPC1 and CPC2). These hydrolysates were evaluated for their antimicrobial activity against Gram-positive (G⁺) and Gram-negative (G^?) microorganisms. The protein hydrolysates were purified by ultrafiltration through a membrane with 3 kDa nominal molecular weight, for evaluation of hypocholesterolemic activity. An inhibition zone was observed when the hydrolysate was tested against S. aureus, and minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) values were obtained. Peptides from chia protein with molecular mass lower than 3 kDa reduced up to 80.7% of 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMG-CoA reductase) enzymatic reaction velocity. It was also observed that, independent of the method used to obtain chia proteins, the fractions showed relevant bioactivity. Moreover, the intensity of the bioactivity varied with the method for obtaining the protein and with the enzyme used in the hydrolysis process. This is the first report to demonstrate that chia peptides are able to inhibit cholesterol homeostasis.     

Modelling the potential distribution of two tropical freshwater fish species under climate change scenarios

1. Climate change is causing shifts in the distribution patterns of freshwater fish at various spatio-temporal scales. Tropical freshwater fish are vulnerable, especially in areas where a high impact of climate change is predicted; thus, there is an increasing need to predict these shifts to determine conservation and adaptation strategies.
2. Ecological niche models offer a reliable way to predict the effects of climate change on species distribution. Potential shifts in the distribution of tropical fish were tested under two scenarios (4.5 – moderate and 8.5 – extreme) with three general circulation models for years 2050 and 2070 using maximum entropy software using as models two predatory species – the tropical gar Atractosteus tropicus and the giant cichlid Petenia splendida.
3. The potential distribution of both species was associated with warm and humid–sub-humid conditions. Future projections showed a higher availability of suitable areas for both species resulting from the expansion of warmer conditions in the middle and upper basins of the Central American mountain range and centre of the Yucatan Peninsula.
4. Ecological niche models of keystone or umbrella species such as A. tropicus and P. splendida could be useful to support conservation plans of protected areas. The potential distribution of both species covers areas of high suitability including six important biosphere reserves in Mexico, three protected areas in Guatemala and part of the Mesoamerican biological corridor.
5. Despite the potential expansion of the present distribution range suggested by the models, it is important to consider the biological and ecological requirements of the species and the ecological implications of these potential shifts in distribution. Both scenarios could have several implications at genetic, population, and ecosystem levels.

     

Influence of Neospora caninum intra-specific variability in the outcome of infection in a pregnant BALB/c mouse model

Previous assays in pregnant animals have demonstrated the effect of different host factors and timing of infection on the outcome of neosporosis during pregnancy. However, the influence of Neospora caninum isolate itself has been poorly investigated. Here, we compared the effects on clinical outcome and vertical transmission observed in a pregnant mouse model following infection with 10 different N. caninum isolates. The isolates in our study included the Nc-Liv isolate and nine N. caninum isolates obtained from calves. Female BALB/c mice were inoculated with 2 × 10⁶ tachyzoites at day 7 of pregnancy. Morbidity and mortality, in both dams and offspring during the course of infection, and transmission to progeny at day 30 postpartum were evaluated. The serum IgG1 and IgG2a production in dams were also examined. All dams showed elevated IgG1 and IgG2a responses, confirming N. caninum infection, although signs of disease were only exhibited in dams infected with 4 of the 10 isolates (Nc-Spain 4H, Nc-Spain 5H, Nc-Spain 7 and Nc-Liv). In neonates, clinical signs were observed in all N. caninum-infected groups, and neonatal mortality rates varied from greater than 95% with the isolates mentioned above to less than 32.5% with the other isolates. Vertical transmission rates, as assessed by parasite PCR-detection in neonate brains, also varied from 50% to 100% according to the isolate implicated. These results confirm the wide pathogenic and transmission variability of N. caninum. The intra-specific variability observed herein could help us explain the differences in the outcome of the infection in the natural host.     

Effect of the chlorinated washing of minimally processed vegetables on the generation of haloacetic acids

Chlorine solutions are usually used to sanitize fruit and vegetables in the fresh-cut industry due to their efficacy, low cost, and simple use. However, disinfection byproducts such as haloacetic acids (HAAs) can be formed during this process, which can remain on minimally processed vegetables (MPVs). These compounds are toxic and/or carcinogenic and have been associated with human health risks; therefore, the U.S. Environmental Protection Agency has set a maximum contaminant level for five HAAs at 60 μg/L in drinking water. This paper describes the first method to determine the nine HAAs that can be present in MPV samples, with static headspace coupled with gas chromatography-mass spectrometry where the leaching and derivatization of the HAAs are carried out in a single step. The proposed method is sensitive, with limits of detection between 0.1 and 2.4 μg/kg and an average relative standard deviation of ～8%. From the samples analyzed, we can conclude that about 23% of them contain at least two HAAs (<0.4-24 μg/kg), which showed that these compounds are formed during washing and then remain on the final product.     

Variability in seedling emergence traits of Patagonian Cypress marginal steppe populations

Genetic information on adaptive traits is crucial for prediction of the evolution of natural populations in relation to global climate change. The seedling emergence process together with germination is a key adaptive stage in any seeding species. We aim to analyze variability in seedling emergence traits within and among marginal populations of Patagonian Cypress (Austrocedrus chilensis (D.Don) Pic. Ser. et Bizzarri), which have been suggested to be of conservation relevance. We performed an emergence trial in a greenhouse with seeds collected from 177 open-pollinated trees from 10 populations. A sigmoidal curve was fitted to the cumulative emergence data (in percentage of the sown seeds) for each replicate of each family. Variability was estimated using ANOVA for six variables: emergence capacity (EC), emergence energy (EE), energy period (EP), emergence initiation (t ₁₀), emergence cessation (t ₉₀) and emergence duration (Dur). The overall trial mean for EC was 76.2 %, while EE was only 27.6 %. Hence, most seedlings emerged after the energy period, which is interpreted as a bet-hedging strategy. Both “population” and “family” factors significantly affected all variables. The proportions of “family” variances were higher than “population” ones for EC, EE, Dur and t ₉₀, but the opposite was found for EP and t ₁₀, which is evidence of differentiation among populations. Variability among families may be due to both genetic and environmental causes, including maternal effects. However, the relatively high proportion of family variability in EC and EE suggests acceptable levels of additive genetic variance, which would not hinder the potential to evolve in these specific traits. Conversely, the chances to adapt in EP and t ₁₀ are lower, and consequently local extinctions driven by global climate change seem possible.