2-Aminoethoxydiphenyl borate (2-APB) reduces alkaline phosphatase release, CD63 expression, F-actin polymerization and chemotaxis without affecting the phagocytosis activity in bovine neutrophils

2-Aminoethoxydiphenyl borate (2-APB) interferes with the Ca(2+) influx and reduces the ROS production, gelatinase secretion and CD11b expression in bovine neutrophils. Moreover, it has been suggested that inhibition of the Ca(2+) channel involved in the store operated Ca(2+) entry (SOCE) is a potential target for the development of new anti-inflammatory drugs in cattle, however it is unknown whether 2-APB affects neutrophil functions associated with the innate immune response. This study describes the effect of 2-APB, a putative SOCE inhibitor, on alkaline phosphatase activity a marker of secretory vesicles, CD63 a marker for azurophil granules, F-actin polymerization and in vitro chemotaxis in bovine neutrophils stimulated with platelet-activating factor (PAF). Also, we evaluated the effect of 2-APB in the phagocytic activity against Escherichia coli and Staphylococcus aureus bioparticles. We observed that doses of 2-APB ≥10 μM significantly reduced alkaline phosphatase activity and in vitro chemotaxis, whereas concentrations of 2-APB ≥50 μM reduced CD63 expression and F-actin polymerization. Finally, we observed that 2-APB did not affect the phagocytic activity in neutrophils incubated with E. coli and S. aureus bioparticles. We concluded that inhibition of Ca(2+) influx could be a useful strategy to reduce inflammatory process in cattle.     

Morphophysiological response of young olive trees to verticillium wilt under different surface drip irrigation regimes

The effects of irrigation on verticillium wilt in olive, in terms of morphological, biomass and physiological parameters were evaluated on pot‐grown trees maintained in the field for 3 years. Plants inoculated and noninoculated with Verticillium dahliae were irrigated to high and low range of soil water content (HR and LR) at daily (DF; about 2 days/event), weekly (WF) and daily‐weekly (DWF) drip‐irrigation frequency. Morphological parameters, relative biomass and biomass water‐use efficiency were higher at LR than at HR (with few exceptions) and at DF than at other frequencies in noninoculated and inoculated plants, but the fungus reduced those parameters by 17.0–38.5%. Lower root weight ratio, relative biomass and shoot length as area originated at HR in noninoculated plants, could be favourable to the accumulation of root infections and the amount of fungus per tissue length in inoculated plants because higher infection was known at HR. Moreover, higher aerial biomass and length promoted by irrigation at DF could prevent the more severe expression of symptoms, which occurred at WF and DWF in the presence of Verticillium. Negative correlations were found between indicated parameters and disease. Lower water stress (S_Ψ), and higher stomatal conductance (g_s) and net photosynthesis at DF in noninoculated plants could limit the disease by improving water status, as S_Ψ was increased by the fungus only at WF and DWF, and g_s and disease were negatively correlated. LR‐DF treatment minimized the disease and kept the growth, water‐use efficiency and physiological parameters in inoculated plants to levels close to noninoculated plants.     

Long-term biological effectiveness of Pochonia chlamydosporia var. catenulata against Meloidogyne incognita infection in a crop rotation systemi

The aim of this study was to determine the effectiveness of Pochonia chlamydosporia var. catenulata and the bacteria Tsukamurella paurometabola strain C-924 (HeberNem?), to control Meloidogyne incognita infection in a crop rotation system (cucumber, tomato and pepper) under greenhouse production during six crop cycles. We determined gall index, prevalence, intensity of disease, and number of juvenile nematodes in soil. At the end of the sixth cycle, plants protected with P. chlamydosporia showed a reduction in gall index (≤2), prevalence, and median (42%) intensity of disease similar or more pronounced than those observed in plants protected with T. paurometabola (80%). Both treatments were more efficient at reducing galling than at reducing prevalence and therefore reduction of intensity of disease was mainly due to the reduction in galling. The hydrolytic activity of P. chlamydosporia and its resistant spores make this fungus one of the best options for the long-term control of RKNs by decreasing egg masses or galls. Pochonia chlamydosporia var. catenulata is an important part of a potential biological control strategy in rotation systems of vegetable crop and can be used for long-time IPM strategies in Cuba and the Caribbean.     

Contribution of lipid oxidation products to acrylamide formation in model systems

The reactions of asparagine with methyl linoleate ( 1), methyl 13-hydroperoxyoctadeca-9,11-dienoate ( 2), methyl 13-hydroxyoctadeca-9,11-dienoate ( 3), methyl 13-oxooctadeca-9,11-dienoate ( 4), methyl 9,10-epoxy-13-hydroxy-11-octadecenoate ( 5), methyl 9,10-epoxy-13-oxo-11-octadecenoate ( 6), 2,4-decadienal ( 7), 2-octenal ( 8), 4,5-epoxy-2-decenal ( 9), and benzaldehyde ( 10) were studied to determine the potential contribution of lipid derivatives to acrylamide formation in heated foodstuffs. Reaction mixtures were heated in sealed tubes for 10 min at 180 degrees C under nitrogen. The reactivity of the assayed compounds was 7 > 9 > 4 > 2 > 8 approximately 6 > 10 approximately 5. The presence of compounds 1 and 3 did not result in the formation of acrylamide. These results suggested that alpha,beta,gamma,delta-diunsaturated carbonyl compounds were the most reactive compounds for this reaction followed by lipid hydroperoxides, more likely as a consequence of the thermal decomposition of these last compounds to produce alpha,beta,gamma,delta-diunsaturated carbonyl compounds. However, in the presence of glucose this reactivity changed, and compound 1/glucose mixtures showed a positive synergism (synergism factor = 1.6), which was observed neither in methyl stearate/glucose mixtures nor in the presence of antioxidants. This synergism is proposed to be a consequence of the formation of free radicals during the asparagine/glucose Maillard reaction, which oxidized the lipid and facilitated its reaction with the amino acid. These results suggest that both unoxidized and oxidized lipids are able to contribute to the conversion of asparagine into acrylamide, but unoxidized lipids need to be oxidized as a preliminary step.     

Identification and relationships of accessions grown in the grapevine (<Emphasis Type="Italic">Vitis vinifera</Emphasis> L.) Germplasm Bank of Castilla y Léon (Spain) and the varieties authorized in the VQPRD areas of the region by SSR-marker analysis

A total of sixty-five accessions from the Spanish region of Castilla y León including those authorized in the VQPRD areas were characterized for six SSR loci. All the samples but one unknown were identified by comparison to other databases. Thirty synonymous samples and three cases of homonymy were found out, confirming in most cases ampelographic expectations. Five unique genotypes belonging to local varieties in risk of extinction were detected. Several parameters were calculated to assess the usefulness of the chosen loci in this work. A dendrogram representing the genetic similarities among the accessions was constructed using the neighbor-joining method to investigate possible parentage relationships in the sample, and to explain them from an historical and cultural point of view.     

Blood magnesium values in healthy cattle and in cattle affected with anaplasmosis and eperythrozoonosis

Magnesium (Mg) concentrations in plasma, serum, and erythrocytes (RBC) of calves subjected to splenectomy (SP), splenectomy-induced eperythrozoonosis (EP), and experimentally induced anaplasmosis (AN) were determined and compared with pre-SP, pre-EP, and pre-AN values. Mean serum and plasma Mg concentrations varied in parallel through most periods of the trial. Both were highest after SP, but neither value was significantly different from the pre-SP concentrations. Mean serum and plasma Mg values varied inversely with mean RBC values after SP and EP. Mean RBC Mg values increased significantly from previous periods during EP and AN and after AN, with the values after AN being significantly higher than those at all other periods. Mean plasma, serum, and RBC Mg concentrations were correlated to both mean percentage of parasitemia (%P) and packed-cell volume (PCV). The RBC Mg values increased significantly after mean %P reached 20 and the PCV began to decrease. There was evidence for a distinct lag variation in increasing RBC Mg concentrations with respect to both mean %P and PCV during AN. A strong correlation was observed between the RBC Mg values and the %P occurring 5 days previously and the PCV occurring 3 days previously.     

A new surgical method for the control of the epiphora in dogs: modified parotid duct transposition

A five-year-old, female, spayed beagle with a history of unilateral epiphora of several months' duration as a result of inadequacy of the lacrimal drainage system was treated successfully with a new surgical method of parotid duct transposition. Data from complete preoperative ophthalmological examination, surgical technique, postoperative treatment, contrast radiography and histological examination were recorded. Absence of complications and complete patency of the stoma at 30 days after surgery confirmed the validity of this new technique. The use of an anatomic duct may be advantageous compared with the current techniques of conjunctivorhinostomy, conjuntivobuccostomy and conjunctival maxillary sinusostomy for the treatment of epiphora in the dogs caused by inadequacy of the lacrimal drainage system.     

Relationship between DNA fragmentation of equine granulosa cells and oocyte meiotic competence after in vitro maturation

The acquisition of equine oocyte developmental capacity is ensured by the follicular environment, such as granulosa cells, which could reflect the meiotic development potential of immature oocytes. This study evaluated the relationship between DNA fragmentation of granulosa cells, using the chromatin dispersion test, and equine oocyte meiotic development after in vitro maturation. Granulosa cells and cumulus–oocytes complexes (n = 50) were recovered from slaughterhouse‐derived ovaries. Oocytes were in vitro matured, stained and evaluated under fluorescence microscopy. Maturation rates were classified into outstanding, medium and poor levels of maturation using 25th and 75th percentiles as thresholds. For DNA assessment, each sample was processed with the Ovoselect^® kit (Halotech DNA). High, low and total DNA fragmentation percentages were compared among levels of maturation rates by ANOVA, followed by Duncan test. Results were expressed as mean ± SE. Total and high DNA fragmentation rates of granulosa cells were significantly higher (p < 0.05) in follicles whose oocytes had reached outstanding maturation level than those originating from follicles whose oocytes had reached poor maturation level. In conclusion, the DNA fragmentation analysis of equine granulosa cells can be a valuable test to identify equine oocytes showing the best meiotic competence after in vitro maturation.     

Assessment of a germplasm bank for the autochthonous cattle breed Asturiana de la Montaña: Extender (Biociphos vs. BIOXCell) affected sperm quality but not field fertility

Semen banking is critical to preserving rare and autochthonous breeds. However, protocols can change with time, leaving heterogeneous semen batches. The objective of this study was to assess differences in sperm quality and field fertility. We report differences between batches frozen with the Biociphos and BIOXCell extenders in the Asturiana de la Montaña cryobank (autochthonous and endangered breed, Northern Spain). Doses from 48 bulls were analysed by CASA and flow cytometry. The 85‐days non‐return rates from AI records were used to assess the fertility of 23,853 AI. BIOXCell showed higher quality post‐thawing. Differences increased after a 5‐hr incubation at 37°C, and Biociphos yielded doses with lower resilience. Field fertility did not differ between extenders (Biociphos: 57.4% ± 1.2; BIOXCell: 56.6% ± 3.0), possibly because of AI protocols compensating for differences in quality. However, this needs to be confirmed by controlled intervention studies. In conclusion, batches frozen with Biociphos may require specific strategies for compensating for the lower sperm quality. Regular surveys and evaluation of cryobank procedures may be useful to characterizing stored batches and defining strategies to guaranteeing success in their future use.