Molecular differentiation of Hypoderma bovis and Hypoderma lineatum (Diptera,Oestridae) by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP)

The most variable region of the cytochrome oxidase I (COI) gene of Hypoderma bovis(1) and Hypoderma lineatum(2) (Diptera, Oestridae) was amplified by PCR and the amplicons were sequenced and analysed. PCR products were digested with three restriction enzymes, namely BfaI, HinfI and TaqI, providing informative profiles. H. bovis and H. lineatum sequences revealed an inter-specific variation rate of 8.5%, and an intra-specific variation rate of 0.87 and 0.29%, respectively. The results showed that the COI gene region examined was useful for the differentiation of H. bovis and H. lineatum and that a PCR-RFLP assay is a practical tool for their identification, offering additional diagnostic and epidemiological instruments for the study of cattle grub infestation.     

Evidence for a relationship between bovine erythrocyte lipid membrane peculiarities and immune pressure from ruminal ciliates

Erythrocytes of bovines and other ruminants have a strikingly anomalous phospholipid composition, with low or absent phosphatidylcholine (PC) together with high sphingomyelin (SM) content. Here, we report the presence in normal bovine serum of high levels of anti-phospholipid antibodies of IgM isotype against, PC and the phosphono analogue of phosphatidylethanolamine, aminoethylphosphonolipid (AEPL), normally produced by rumen ciliates. In contrast, no antibodies were detected against SM or N-acyl-phosphatidylethanolamine (NAPE), the major components of bovine erythrocytes. In addition, we found that exposure of the ciliate Tetrahymena thermophila to bovine serum results in rapid lysis, an effect that was inhibited by adsorption of the serum with SM/AEPL liposomes. Furthermore, incubation with bovine serum had a similar effect on freshly obtained ruminal ciliates, and the lytic activity was eliminated by pre-adsorption of the serum with SM/PE liposomes. The ruminant mode of life with its concomitant ciliate fauna is hereby linked to the peculiar conformation of bovine erythrocyte membranes. We propose that the unique phospholipid composition of bovine erythrocytes appears as an evolutionary adaptation to tolerate the lytic effects of anti-phospholipid antibodies generated against AEPL, a membrane component of the huge mass of ruminal ciliates, necessary commensals of this group of mammals.     

Diversity in phosphorus mobilisation and uptake in ectomycorrhizal fungi

• Introduction   

Phosphorus (P) is often the first or second element limiting aboveground net primary productivity of forests. Besides low available inorganic orthophosphate (Pi) concentrations, soil may contain high total P contents, as insoluble mineral P or as organic P. Most plants form mycorrhizal associations that improve their P nutrition. Three main hypotheses have been proposed to explain this positive effect through an increase of (1) P mobilisation from mineral P, (2) P mobilisation from organic P and (3) soil exploration and P uptake. However, the positive effect of mycorrhizal symbiosis may be variable with the fungal species forming the association. This could be due to the different abilities of mycorrhizal fungi to mobilise P and/or to take up Pi from the soil.     

Efficacy of alpha-cyclodextrin against experimental cryptosporidiosis in neonatal goats

The efficacy of orally administered tablets containing alpha-cyclodextrin, an excipient used in the pharmaceutical industry with demonstrated anticryptosporidial activity in vitro and in neonatal mice, was evaluated in neonatal goat kids. The formulation was evaluated for hardness and was subjected to in vitro drug release studies. Twenty goat kids were orally inoculated with 10(6) oocysts of C. parvum within the first 6 days of age. Half of the animals were treated by oral administration of four tablets of alpha-cyclodextrin/day (500 mg/kg of body weight) for six consecutive days, the treatment beginning on the day of inoculation. Infection was monitored by daily examination of faecal samples from the first day to 25 days post-inoculation. The criteria studied in evaluating efficacy were: oocyst shedding, presence of diarrhoea and weight gain at 15 and 25 days post-inoculation. alpha-cyclodextrin was effective when given at the beginning of infection: there was a longer pre-patent period, a reduction in the patent period and a decrease in the intensity of infection, these differences being statistically significant (P < 0.05) compared with untreated neonatal kids. Moreover, except in one animal, the diarrhoea was prevented in infected neonatal kids. Animals from both groups increased the body weight and no significant differences were seen between the two groups.     

Chemopreventive activity of polyphenolics from black Jamapa bean (Phaseolus vulgaris L.) on HeLa and HaCaT cells

The antiproliferative effects of 100% methanol crude extract and of Toyopearl and silica gel fractions from the seed coats of black Jamapa beans (Phaseolus vulgaris L.) were evaluated using HeLa, human adenocarcinoma cells, and HaCaT, human premalignant keratinocytes. The 100% methanol crude extract [172.2 microM equiv of (+)-catechin] increased adhesion of HeLa cells; however, 3- and 5-fold higher concentrations decreased the number of cells attached as a function of the treatment time. The highest concentration tested diminished the cell adhesion until 40% (after 24 h) to almost 80% (after 72 h). The IC50 values showed that the 100% methanol crude extract was the most effective inhibitor of HeLa cell proliferation, even when it was dissolved in dimethylsulfoxide (DMSO) [34.5 microM equiv of (+)-catechin] or in medium [97.7 microM equiv of (+)-catechin]. The Toyopearl 5 (TP5) fraction and silica gel 2 (SG2) fraction inhibited 60% of the HeLa cell proliferation. The IC50 was 154 microM equiv of (+)-catechin of the 100% methanol crude extract on HaCaT cells. Toyopearl fractions TP4 and TP6 significantly inhibited HaCaT cell proliferation, but the silica gel fractions did not have a significant effect. The 100% methanol crude extract (35 microg of dry material/mL) decreased the number of HeLa cells in the G0/G1 phase from 68.9% (for control cells) to 51.4% (for treated cells) and increased apoptosis (2.9 and 21.2% for control and treated cells, respectively). The results indicated that black Jamapa beans could be a source of polyphenolic compounds, which have an inhibitory effect toward HeLa cancer cells but are less aggressive on HaCaT premalignant cells.     

Bovine herpesvirus 4-associated postpartum metritis in a Spanish dairy herd

In more than 10 Spanish dairy cows, a bovine herpesvirus 4 (BHV4) associated postpartum metritis was confirmed by virus isolation, BHV4-glycoprotein B (gB) PCR and/or serology. In this study, 12 cows with, and, at the time of sampling, 3 cows without clinical signs of acute postpartum metritis from one large dairy herd in Spain were examined for bacterial and viral infections. Blood, placenta/caruncles and uterine contents were collected between day 1 and day 20 post-calving, and examined for the presence of bacteria and for viruses by virus isolation, BHV4 DNA by BHV4-gB PCR and/or BHV4 antibody titres. Bovine herpesvirus 4 was detected in 83% of the cases with clinical signs of acute postpartum metritis by virus isolation and/or BHV4-gB PCR. An increase of BHV4 antibodies was detected in all examined postpartum metritis cows and in the 3 cows without clinical metritis. Two of these 3 cows developed severe metritis a few dayss after collecting the first blood sample. A concurrent infections of BHV4 and bacteria, mainly Arcanobacterium pyogenes and Streptococcus sp., were detected in 73% of the examined uterine contents collected from postpartum metritis affected cows. This case-report study showed a clear association between BHV4 infections and acute postpartum metritis in dairy cows. In addition, the BHV4-associated postpartum metritis appeared to be an emerging syndrome in this Spanish herd.     

Enhancement of alkalinity tolerance in two cucumber genotypes inoculated with an arbuscular mycorrhizal biofertilizer containing Glomus intraradices

The aim of the present study was to determine whether arbuscular mycorrhizal (AM) inoculation with a biofertilizer containing clays as granular carriers, leek root pieces and Glomus intraradices spores could improve alkalinity tolerance of two cucumber genotypes, and to study the changes induced by AM at agronomical and physiological level. A greenhouse experiment was carried out to determine yield, growth, fruit quality, net photosynthesis (A_CO2), electrolyte leakage, and mineral composition of two cucumber (Cucumis sativus L.) genotypes (hybrid “Ekron” or open-pollinated variety “Marketmore”) with inoculated and noninoculated arbuscular mycorrhizal biofertilizer. Plants were supplied with nutrient solutions at two pH values (6.0 or 8.1). The high pH nutrient solution had the same basic composition with an additional 10 mM NaHCO₃ and 0.5 g l⁻¹ CaCO₃. The percentage root colonization was higher in “Marketmore” (21.8%) than “Ekron” (12.7%). Total and marketable yield and total biomass were significantly higher by 189%, 213%, and 77%, respectively, with Ekron in comparison to those recorded with Marketmore. The highest crop performance with Ekron in comparison to Marketmore was due to the improved nutritional status (higher N, P, K, Ca, Mg, Fe, Mn, and B), higher leaf area, and net photosynthesis. Increasing the concentration of NaHCO₃ from 0 to 10 mM in the nutrient solution significantly decreased yield, plant growth, A_CO2, N, P, Fe, Cu, Zn, Mn, and B concentration in leaf tissue, whereas the electrolyte leakage increased. The inoculated plants under alkaline conditions had higher total, marketable yield, and total biomass than noninoculated plant. Mycorrhizal cucumber plants grown under alkaline conditions had a higher macronutrient concentration in leaf tissue compared to noninoculated plants. The highest yield and biomass production in inoculated plants seems to be related to the capacity of maintaining higher net A_CO2 and to a better nutritional status (high P, K, Mg, Fe, Zn, and Mn and low Na accumulation) in response to bicarbonate stress with respect to −AM plants.     

Assessing foraging strategies of herbivores in Mediterranean oak woodlands: a review of key issues and selected methodologies

Montados are agro-silvo-pastoral ecosystems, typical of the Southwest Iberian Peninsula, of high socio-economic and conservation importance, where grazing is a dominant activity. Montados are characterized by an open tree canopy of Quercus sp. and a diverse undercover of shrubs and grasslands that constitute the plant food resources for grazing animals. Plant food resources of Montados are highly variable, both spatially and seasonally, in quantity and quality. Reliable and easy to use methods to monitor grazing are necessary to allow proper understanding of foraging strategies of grazing animals and to set sustainable grazing management. We describe the main characteristics of the plant food resources available for grazing animals, striking its variability, and revise the potential of using N-alkanes and saliva proteome methods to assess foraging strategies in Montados. In a scenario of dynamic multiple choices, we discuss the use of n-alkane methodology for the simultaneous estimation of diet composition and voluntary intake and saliva proteome as a mean of increasing the knowledge on diet adjustments.     

The effect of four calcium‐based amendments on soil aggregate stability of two sandy topsoils

The structural stability of soil is a physical characteristic that affects soil degradation processes. Calcium‐based amendments, such as calcium carbonate, calcium sulfate, and calcium oxide/hydroxide, have been shown to improve the stability of soil aggregates. This study seeks to determine which calcium‐based soil amendments, and at what concentration, are the most efficient in improving aggregate stability of sandy topsoils derived from granitic and metamorphic parent materials, and to analyze the mechanisms involved. In the pot experiment, soils amended with CaCO₃, CaCl₂, and CaSO₄ did not present significant differences in aggregate stability compared to the control or among each other. In contrast, Ca(OH)₂ soil amendment brought the greatest stability to the soil aggregates. A dose of 1% Ca(OH)₂ significantly increased the stability of soil aggregates. This effect is due to the reaction of Ca(OH)₂ with atmospheric CO₂ which leads to the formation of CaCO₃, a delayed reaction not showed by the other soil amendments tested. Likewise, the greater solubility of Ca(OH)₂ compared to CaCO₃ exerts a greater aggregation effect on soil. Thus, the mechanism of action of Ca(OH)₂ is related to cementation, rather than flocculation. Future studies should be carried out to demonstrate the effectiveness of Ca(OH)₂ under field conditions.