Analysis of In vitro Fertilizing Capacity to Evaluate the Freezing Procedures of Boar Semen and to Predict the Subsequent Fertility

A porcine in vitro fertilization (IVF) system and seminal quality parameters of frozen–thawed boar semen were used to assess the effectiveness of two different thawing rates of frozen boar semen, and to address the question of whether differences between fertility of ejaculates could be predicted in a limited field trial. In the first experiment, two thawing procedures were analysed (37°C, 30 s; 50°C, 12 s) and no differences in sperm quality were found. However, when the procedure was 50°C, 12 s the IVF results showed a higher number of sperm per penetrated oocyte and a near 10 points higher rate of pronuclear formation. In the second experiment, the fertility results obtained in the limited field trial show to be efficient enough for application in a commercial use, especially for three of the employed boars (fertility ≥80%). In this limited study, the conventional seminal parameters are not accurate enough to discriminate good and bad boars in relation to fertility. On the contrary, parameters of in vitro penetrability are more precise to predict subsequent fertilities. As conclusion, the IVF fertilization system seems to be a good tool to evaluate the quality of frozen–thawed boar semen previous to its commercial way, to verify the bank semen storage quality and a good way to assay new sperm freezing procedures, as it is the more precise evaluating method in estimating the potential fertilizing ability.     

Evaluation of a Killed Vaccine Against Porcine Pleuropneumonia Due to Haemophilus pleuropneumoniae

A bacterin containing serotypes 1 and 5 of Haemophilus pleuropneumoniae was developed for the prevention and the control of porcine pleuropneumonia. It was injected intramuscularly into three groups of ten piglets, the first group with one dose, the second one with two doses and the third one with three doses at two-week intervals. Another group of ten piglets did not receive the vaccine. All the piglets were then challenged by an aerosol of mixed suspensions of H. pleuropneumoniae serotypes 1 and 5. Two and three injections of vaccine completely prevented mortality, whereas half of the control piglets and of those receiving only one dose of vaccine died. All surviving piglets, both control and vaccinated, had severe signs of respiratory disease for at least 36 hours after exposure to challenge. Moreover, vaccination did not induce the production of antibodies at high titers. Local reactions were not noted after vaccination and at postmortem; ten weeks after the challenge, there were no signs of abscess formation or induration.     

PCR Assay for Identification of Aspergillus Carbonarius and Aspergillus Japonicus

Black Aspergilli, and in particular Aspergillus carbonarius, are the main causes of contamination of grapes and their by-products by ochratoxin A. A PCR-based method was developed to detect DNA of A. carbonarius and A. japonicus. Two pairs of primers (CARBO1/2 and JAPO1/2) designed from the calmodulin gene, produced PCR products of 371 and 583 bp for A. carbonarius and A. japonicus, respectively. Primer specificity was tested with DNA of 107 strains belonging to Aspergillus section Nigri isolated mostly from grapes in Europe. The sensitivity of primers CARBO1/2 and JAPO1/2 was 12.5 pg when using pure total genomic DNA of the two species. The developed primers provide a powerful tool for detection of the main ochratoxigenic producing Aspergillus species in grapes.     

The antihistaminic chlorpheniramine inhibitsin vitro growth of several fungi isolated from harvested fruits

Chlorpheniramine (CPA) is an antihistaminic that changes the conformation of DNA and inhibits polyamine biosynthesis in mammalian cells. In the present work, we tested the effect of CPA on four genera of fungi species (Altemaria alternata, Botrytis cinerea, Cladosporium cladosporioides and threePenicillium spp.) grownin vitro. Similar growth inhibitions of these genera were produced by 0.5 raM iprodione, CPA and histidinol, but CPA was the most effective. The CPA sensitivities of the twoB. cinerea strains were different. Putrescine did not restore the fungal growth inhibited by CPA.     

Isolation and pathogenicity of Colletotrichum spp. causing anthracnose of strawberry in south west Spain

Anthracnose, caused by Colletotrichum spp., is a major disease of cultivated strawberry, Fragaria × ananassa. This study identifies the Colletotrichum spp. which causes strawberry anthracnose in the southwest of Spain. A survey of the region was carried out, and the strains isolated were identified as C. acutatum by using the polymerase chain reaction (PCR) with genus and species-specific primers, demonstrating that this species is currently the causal agent of strawberry anthracnose in the studied region. The pathogenicity of C. acutatum and C. gloeosporioides strains was evaluated on two principal strawberry cultivars (cvs Camarosa and Ventana) under field conditions, the latter being more pathogenic than the former.     

Distribution and diversity of leaf-cutting ants in Northeastern Argentina: species most associated with forest plantations

Abstract

Leaf-cutting ants (LCA) are considered one of the main herbivores and one of the most destructive pest insects of the Neotropics. Northeastern Argentina harbors the greatest species richness of these ants and in turn comprises the highest surface with forest plantations. Our aim was to establish which species of leaf-cutting ants are most commonly associated with forest plantations by analyzing their geographic distribution using published and unpublished species occurrence data. Also, estimate their potential areas of distribution along a latitudinal gradient that entirely encompasses northeastern Argentina using Ecological Niche Modeling. Only seven of the 20 species recorded were strongly associated with productive systems along the gradient, but only 2–3 species in each region could be considered high-risk species for forest plantations. High-risk species composition shows a turnover between regions. Our models show the potential distribution areas where LCA could become more abundant and dominant, and possibly causing a detrimental effect on the forest plantations in the studied region. We find that ecological niche models are useful tools to assess the environmental suitability of important LCA.     

The use of Verticillium dahliae and Diplodia scrobiculata to induce resistance in Pinus halepensis against Diplodia pinea infection

The pathogenic fungi Verticillium alboatrum and Diplodia scrobiculata were assayed for biological control of Diplodia pinea on Aleppo pine (Pinus halepensis) in Catalonia (north-eastern Spain). Young shoots were pre-treated with inoculations of either V. dahliae or D. scrobiculata, by placing colonized agar plugs on wounds made by removing a single needle fascicle. An inoculation with D. pinea was performed 15 days later. Two months after the shoot inoculations, the canker length on the stems was measured and the percentage of shoot dieback calculated. Verticillium dahliae and D. scrobiculata were found to significantly reduce the canker length of D. pinea (P < 0.05) when compared with positive controls. Diplodia pinea was slightly more sensitive to V. dahliae than to D. scrobiculata, but no significant differences (P > 0.05) were observed in the mean canker length between the two treatments. Trees pre-inoculated with V. dahliae resulted in 31.12% shoot dieback, while those pre-inoculated with D. scrobiculata resulted in 32.18% shoot dieback, compared with positive controls (42.85%).     

Storage performance of clementines treated with hot water,sodium carbonate and sodium bicarbonate dips

Summary

The potential of using hot water (2.5 min at 45°C), 2% sodium carbonate or 2% sodium bicarbonate solutions alone or combined with hot water, for control of Penicillium digitatum (green mould) was investigated on commecially ripe clementines. The effect of such treatments on the post-harvest storage life of clementines was determined during two months analysing weight loss, increase in rind deformation and internal maturity. Carbonate and bicarbonate solutions effectively control green mould during storage but hot water does not. Compared with the control and hot-water treated fruits, clementines treated with carbonate and, to a lesser extend with bicarbonate, showed higher weight loss and lower firmness during storage. In contrast, lower increase in ripening index was found when the fruits were treated with carbonate or bicarbonate. Fruits treated with hot water alone or combined with carbonate exhibited no change in ripening during the first month of storage. Rinsing the fruit significantly reduces weight and firmness loss in the fruits treated with carbonate and bicarbonate, but did not change the ripening index. The primary finding of this work was that carbonate and bicarbonate treatments might significantly increase the loss in weight and firmness during storage. Rinsing the fruit immediately after treatment appeared to be a simple solution to avoid these losses in quality.