Cumulus–Oocyte Communications in the Horse: Role of the Breeding Season and of the Maturation Medium

Horse is a seasonal breeder and information on oocyte quality outside the breeding season is very limited. Ovaries obtained at the slaughterhouse are a convenient but often limited source of oocytes in this species. As the low quantity of ovaries leads to an intensive use of all available material, it would be useful to know whether ovaries collected during the non‐breeding season are suitable for in vitro maturation (IVM). In an attempt to characterize the effect of season on oocyte quality, we investigated the permeability of the gap junctions (GJ) present between cumulus cells and oocytes because of their important role in oocyte growth and maturation. We also compared the effect of supplementing the maturation medium with bovine serum albumin (BSA) or oestrus mare serum (EMS). A total of 645 oocytes isolated from 158 and 154 ovaries collected during the breeding and the non‐breeding season, respectively, were used in this study. Oocytes were matured for 30 h in TCM 199 supplemented either with 10% EMS or with 4 mg/ml BSA. The presence of permeable GJs between cumulus cells and oocytes was investigated with the injection of a 3% solution of the fluorescent dye Lucifer yellow into the ooplasm. No differences in efficiency of oocyte retrieval or oocyte meiotic competence were detected between oocytes collected during the breeding and non‐breeding season. The vast majority (90%) of the oocytes collected during the breeding season had fully functional communications with their surrounding cumulus cells but such communications were completely interrupted in 55.3% of the oocytes collected during the non‐breeding season. During the non‐breeding season, the proportion of oocytes whose communications with cumulus cells were classified as closed or intermediate at the end of maturation was lower in the group matured with BSA than with EMS (71.4 vs 97.7, p < 0.05). The same trend, although not statistically significant, was observed during the breeding season also. The presence of BSA caused an incomplete cumulus expansion during both seasons. Our data indicate that oocytes collected during the non‐breeding season do not show any meiotic deficiency but lack active communication with the surrounding cumulus cells at the time of their isolation from the ovary. No data are available at present for determining the consequences on the developmental competence even if data from other species suggest that this is likely.     

p53 protein expression in conjunctival squamous cell carcinomas of domestic animals

The expression of p53 protein was investigated in eight formalin-fixed, paraffin-embedded conjunctival squamous cell carcinomas of five horses and one cow, dog and cat each by an immunohistochemical procedure in order to evaluate protein overexpression. Anti-human p53 protein mouse monoclonal antibodies known to be cross-reactive with p53 protein of the animal species examined were used. Positive p53 nuclear immunostaining was detected in five equine, one bovine and one feline cases. Conversely, no p53 immunostaining was found in the only canine case examined. These results demonstrate a frequent p53 overexpression in conjunctival squamous cell carcinoma that could be related to UV-induced mutations of the p53 tumor suppressor gene.     

Persistence and translocation of a benzothiadiazole derivative in tomato plants in relation to systemic acquired resistance against Pseudomonas syringae pv tomato

A reproducible and accurate procedure, based on HPLC analysis, has been developed to determine simultaneously acibenzolar‐S‐methyl (CGA 245 704) and its acid derivative (CGA 210 007) in tomato leaves. The limit of detection and quantification of the method are 0.015 and 0.15 mg litre^?1 for CGA 245 704 and 0.030 and 0.30 mg litre^?1 for CGA 210 007. In tomato plants treated with 250 µM CGA 245 704, it was found that the inducer rapidly translocates from treated leaves (cotyledons, 1st and 2nd) to untreated leaves (3rd to 5th), with the maximum translocation (40% of the total quantity found) occurring 8 h after the treatment. CGA 245 704 residues decreased as time elapsed in both treated and untreated tomato leaves, reaching negligible values 72 h after treatment. The acid derivative, CGA 210 007, was formed in tomato plants as early as 2 h after CGA 245 704 treatment, albeit only in the treated leaves. CGA 210 007 residues decreased in treated tomato leaves with a trend similar to that observed for CGA 245 704. Treatment of tomato plants with CGA 245 704 or CGA 210 007 at 250 µM systemically protected the plants against Pseudomonas syringae pv tomato attacks, the causal agent of bacterial speak disease. Evidence of this were reductions in the degree of infection, the bacterial lesion diameter and the bacterial growth in planta. Since neither CGA 245 704 nor CGA 210 007 inhibited bacterial growth in vitro and the protection against bacterial speak of tomato was observed when the two compounds were completely degraded, the protection must be due to the activation of the plant's defence mechanisms. © 2001 Society of Chemical Industry     

Forage and animal production on palisadegrass pastures growing in monoculture or as a component of integrated crop–livestock–forestry systems

To meet the global demand for animal protein, sustainable intensification of existing livestock systems may be possible, especially through integration of livestock with crops or forestry. Thus, our objective was to compare forage production and animal performance in grass monoculture and integrated systems in the Brazilian Amazon biome. The four systems were (a) livestock (L) with Marandu palisadegrass {Brachiaria brizantha (Hochst. ex A. Rich.) R. D. Webster} as monoculture, (b) palisadegrass pastures integrated with eucalyptus trees (Eucalyptus urograndis; hybrid of Eucalyptus grandis W. Hill ex Maiden and Eucalyptus urophylla S. T. Blake) arranged in three‐row groves with groves spaced 30 m apart (livestock–forestry; LF), (c) palisadegrass after two years of crops (crop–livestock; CL) and (d) palisadegrass after two years of crops with single rows of eucalyptus trees spaced 37 m apart (crop–livestock–forestry; CLF). From July 2016 to July 2017, all experimental units were continuously stocked using a variable stocking rate. Greater herbage accumulation (HA) occurred in CL and CLF in comparison with L and LF (21,310, 24,050, 19,500 and 18,890 kg DM/ha respectively). The gain per hectare of L, LF and CL (average of 932 kg ha^–1) was less than CLF (1,190 kg ha^–1). Average daily gain was similar among systems (0.69 kg/day). We conclude that integrated systems can support similar (LF or CL) or greater (CLF) levels of animal production than palisadegrass monocultures while increasing diversity of outputs, thereby providing a greater range of viable systems for livestock production in the Brazilian Amazon biome.     

Samples disturbance overestimates phosphorus adsorption capacity in soils under long-term application of pig slurry

Grinding soil for analysis causes the destruction of aggregates and may expose adsorption sites that would not be available for adsorption in the soil with preserved structure. The study aimed to evaluate the adsorption of P in three soils with and without preserved structure, submitted to pig slurry (PS) application for a long time. Three soils (Acrisol, Ferralssol and Nitisol) with contrasting textures and P contents, with and without PS were selected and samples were collected with and without preserved structure in the 0–10 and 10–20 cm layers. The undisturbed soil columns were submitted to continuous flow, for 10–24 h, of 300 mg L^?1 P solution and, at regular intervals, aliquots of the resulting solution were collected and the P content was determined. These data were adjusted exponential models to estimate the maximum adsorption of P. The maximum capacity of P adsorption of the soils were 1.4 to 14 times higher in the milled soil samples, making it clear that the destruction of the soil structure, and consequently the porosity and the preferential paths of percolation of the liquids in the soil leads to an overestimation of the soil capacity to adsorb the P applied to the surface.     

Induction of wheat and maize glutathione S-transferase by some herbicide safeners and their effect on enzyme activity against butachlor and terbuthylazine

The expression of glutathione S-transferase (GST) activity in wheat and maize shoots was investigated in response to treatments with the herbicide safeners benoxacor, cloquintocet-mexyl, fenchlorazole-ethyl, fenclorim, fluxofenim and oxabetrinil. These safeners significantly enhanced the GST activity towards 1-chloro-2,4-dinitrobenzene (CDNB) as a 'standard' substrate, with the exception of oxabetrinil in maize. The enhancements of GST (CDNB) activity were found to be concomitant with increases in V(max) (the reaction rate when the enzyme is fully saturated by the substrate) in wheat following cloquintocet-mexyl and fenchlorazole-ethyl treatments, and in maize following fenchlorazole-ethyl treatment. Otherwise, decreases in V(max) were observed in wheat and maize following fenclorim and fluxofenim treatments. With the exception of oxabetrinil, all the safeners significantly reduced the apparent K(M) (the substrate concentration required for 50% of maximum GST activity) of both wheat and maize GST. The V(max) and K(M) variations following safener treatments are discussed in terms of an increased expression of GST enzymes and an increased affinity for the CDNB substrate. The activity of wheat and maize GST was also assayed towards butachlor and terbuthylazine respectively; the results indicate the ability of cloquintocet-mexyl, fenchlorazole-ethyl and fluxofenim to enhance the enzyme activity in wheat and of benoxacor and fenchlorazole-ethyl to do so in maize.     

Evaluation of changes in microbiota after fecal microbiota transplantation in 6 diarrheic horses

The purpose of this study was to characterize the fecal microbiota of horses with acute and chronic diarrhea before and after fecal microbiota transplantation (FMT). Six client-owned horses with acute and chronic diarrhea received FMT from 2 healthy donor horses. Microbiota analysis using next-generation sequencing was performed on fecal samples collected before and 2 and 7 d after FMT. Signs of diarrhea improved in 4 horses, whereas the remaining 2 horses did not survive. There was a significant difference in the number of bacterial species between donors and recipients (P < 0.05). The Order Lactobacillales and the genera Lactobacillus, Intestinimonas, and Streptococcus were increased in the microbiota of diarrheic horses, and Saccharofermentans genus increased in healthy donors. The results suggest that FMT from the healthy donors was not effective over a 7-day period as it did not change the fecal microbiota of the diarrheic horses. Further research to improve the efficacy of FMT in horses is needed.     

Evaluation of a phenological model for strategic decisions for hemp (Cannabis Sativa L.) biomass production across European sites

The optimal combination of yield and quality of hemp fibres from field grown crops is around flowering. Therefore prediction of flowering time would support in planning production and optimization of the cultivar choice for different agro-ecological zones. In the current paper the validation of a recently published model (Amaducci et al., 2008a) is carried out for four varieties across a wide range of sites and thus of air temperature-photoperiod combinations. The model was evaluated by comparing its output to field observations of the duration between emergence and 50% flowering. The model output and observed times from emergence to 50% of flowering generally corresponded well, but some discrepancies were apparent. The biggest discrepancies between estimates and actual data were observed at extreme latitudes. The level of accuracy of the model predictions is satisfactory for strategic decision regarding sowing and harvesting time and cultivar choice, but tactical decisions (e.g. time of harvest based on flowering time) cannot be accurately supported.