A Scanning Electron Microscopy Study of Frozen / Thawed Dog Sperm During In Vitro Gamete Interaction     

J Palomino  M De los Reyes 《Reproduction in domestic animals》2009,44(2):278-283

The aim of this work was to study the effects of cryopreservation on the binding and penetration of dog spermatozoa to the zona pellucida (ZP) by scanning electron microscopy (SEM). The sperm-rich fraction of six ejaculates from five dogs was divided into two aliquots and washed by centrifugation. One aliquot was processed as fresh control sample and the other aliquot frozen in Tris-fructose extender. Gamete interaction was assessed using in vitro matured bitch oocytes, which were co-incubated for up to 3 h. At hourly intervals after the start of co-incubation, in vitro fertilized (IVF) oocytes were processed by SEM. The results were analysed statistically using the anova test. Differences in binding and penetration of the spermatozoa to the ZP occurred; a lower proportion of oocytes with spermatozoa bound to ZP was observed using frozen sperm (p < 0.05) than with fresh sperm (61%, 57% and 53% vs 42%, 40% and 44% at 1, 2 and 3 h, respectively). The percentage of ZP penetration by fresh sperm was directly proportional to the time of co-incubation (9%, 25% and 34%; p < 0.05); in contrast, no differences were observed in the penetration rate with frozen-thawed sperm (21%, 17% and 21%). More acrosome reacted sperm were observed in frozen sperm than in fresh sperm on the surface of the ZP. The differences in the percentage of binding and penetration between fresh and frozen sperm during the co-culture could indicate that the time course of penetration is faster in frozen-thawed dog spermatozoa than in fresh sperm, but that fresh spermatozoa can penetrate more oocytes over a given period of time, which may be related to their reacted or non-reacted initial status.  相似文献   

A combination of Lactobacillus buchneri and Pediococcus pentosaceus extended the aerobic stability of conventional and brown midrib mutants–corn hybrids ensiled at low dry matter concentrations by causing a major shift in their bacterial and fungal community     

Juan J Romero  Jin Park  Younghoo Joo  Yuchen Zhao  Marjorie Killerby  Diana C Reyes  Francesco Tiezzi  Eduardo Gutierrez-Rodriguez  Miguel S Castillo 《Journal of animal science》2021,99(8)

We evaluated the effects of applying a combination inoculant to four corn hybrids harvested at high moisture on their nutritive value and microbial populations. The treatment design was the factorial combination of corn hybrids ensiled with (INO) and without (CON) inoculant. The hybrids were TMF2R737 (MCN), F2F817 (MBR), P2089YHR (PCN), and PI144XR (PBR), ensiled at dry matter (DM) concentrations of 30.5%, 26.3%, 31.1%, and 31.5%, respectively; MBR and PBR were brown midrib mutants (BMR). The inoculant contained Lactobacillus buchneri and Pediococcus pentosaceus (4 × 10⁵ and 1 × 10⁵ cfu/g of fresh corn). The experiment had a complete randomized design with treatments replicated six times. Corn was treated or not with inoculant, packed into 7.6 L bucket silos, and stored for 100 d. At d 0, the relative abundance (RA, %) of Enterobacteriaceae was lower in PBR vs. the other hybrids [51.3 vs. x¯ = (average of) 58.4] and in the case of fungi, incertae sedis (i.s.) Tremellales and Mucoraceae were more and less abundant, respectively, in conventional hybrids vs. BMRs (x¯= 25.8 vs. x¯ = 13.9 and x¯ = 3.64 vs. x¯ = 7.52; P < 0.04). After ensiling, INO had higher LAB (9.3 vs. 7.1 log cfu/g of fresh corn) and acetic acid (3.44% vs. 1.32% of DM) and lower yeast (3.1 vs. 4.6) and molds (1.5 vs. 3.0), and also extended the aerobic stability (582 vs. 111 h) but decreased DM recovery (95.6% vs. 97.4%) vs. CON (P < 0.02). Inoculation reduced bacterial phylogenetic diversity (6.75 vs. 14.4) but increased fungal observed taxonomical units (46 vs. 20) vs. CON (P < 0.01). Also, a higher relative abundance (RA) for Lactobacillaceae (99.2% vs. 75.7%) and lower for Enterobacteriaceae (0.28 vs. 9.93) was observed due to inoculation (P < 0.001). For fungi, INO had a lower RA compared to CON for Monascaceae (12.6 vs. 44.7) and increased i.s. Tremellales (8.0 vs. 1.2) and i.s. Saccharomycetales (6.4% vs. 0.3%; P < 0.006). Inoculation changed the diverse bacterial community found in the phyllosphere across hybrids to a taxonomically uneven one dominated by Lactobacillaceae. In the case of fungi, INO application increased the fungal diversity at d 100 mainly by reducing the dominance of Monascaceae vs. CON. In conclusion, the INO treatment overwhelmed the disparate microbial populations found across BMR and conventional hybrids ensiled at low DM concentrations and ensured a significant concentration of acetic acid that modified fungal populations and in turn extended the aerobic stability of all hybrids.  相似文献   

Application of isotope dilution analysis for the evaluation of extraction conditions in the determination of total selenium and selenomethionine in yeast-based nutritional supplements   总被引：1，自引：0，他引：1  

Hinojosa Reyes L  Marchante-Gayón JM  García Alonso JI  Sanz-Medel A 《Journal of agricultural and food chemistry》2006,54(5):1557-1563

Isotope dilution analysis (IDA) has been used to quantify total selenium, total solubilized selenium, and the selenomethionine (SeMet) amount in yeast and yeast-based nutritional supplements after acid microwave digestion and different enzymatic extraction procedures. For this purpose, both a (77)Se-enriched SeMet spike, previously synthesized and characterized in our laboratory, and a (77)Se(VI) spike were used. In the analysis of the nutritional supplements, the SeMet spike was added to the sample and extracted under different conditions, and the (78)Se/(77)Se and (80)Se/(77)Se isotope ratios were measured as peak area ratios after high-performance liquid chromatography (HPLC) separation and inductively coupled plasma mass spectrometry (ICP-MS) detection. The formation of SeH(+) and mass discrimination were corrected using a natural SeMet standard injected every three samples. Similarly, total solubilized selenium was measured in the extracts after enzymatic hydrolysis using the (77)Se-enriched SeMet as a spike by direct nebulization without a chromatographic separation. To establish a mass balance, total selenium was also determined by IDA-ICP-MS on the yeast tablets after microwave digestion using (77)Se(VI) as a spike. Results showed that all enzymatic procedures tested were able to solubilize total selenium quantitatively from the solid. However, the recovery for the species SeMet, the major selenium compound detected, was seriously affected by the enzymatic procedure employed and also by the matrix composition of the supplement evaluated. For the yeast sample, SeMet recovery increased from 68 to 76% by the combined use of driselase and protease. For the nutritional supplements, the two most effective procedures appeared to be protease and driselase/protease, with a SeMet recovery ranging from 49 to 63%, depending upon the supplement evaluated. In the case of in vitro gastrointestinal enzymolysis, the results obtained showed 26-37% SeMet recovery, while the rest of selenium was solubilized as other unknown compounds (probably Se-containing peptides).  相似文献   

Spice crops agroforestry systems in the East Usambara Mountains,Tanzania: growth analysis   总被引：1，自引：0，他引：1  

T. Reyes  R. Quiroz  O. Luukkanen  F. de Mendiburu 《Agroforestry Systems》2009,76(3):513-523

A scarcity of cultivation land calls for more intensive and productive land use in the East Usambara Mountains in NE Tanzania. Spice crops could generate cash in higher parts of the mountains, but the present cultivation methods are depleting the valuable forest resources. The trial was established at the end of 2000 to find out how the two popular cash crops, cardamom (Elettaria cardamomum (L.) Maton.) and black pepper (Piper nigrum L.), normally grown under the natural forest, will produce in intensive agroforestry system with two multipurpose farm trees, Grevillea robusta A.Cunn. and nitrogen fixing Gliricidia sepium Jacq. Results from 6 years showed that cardamom produced better with grevillea than in natural forest; 5.5 times more in the fourth year than the average in the area. The Land Equivalent Ratios for black pepper and cardamom showed that pepper intercropped with grevillea produced 3.9 times more than in monoculture whereas cardamom intercropped with grevillea and pepper produced 2.3 times more than in monoculture. Gliricidia improved the nitrogen and organic matter content of the soil over the levels found in natural forest. Soil acidity was, however, preventing the plants from using the available mineral nutrients more effectively.  相似文献   

Resistance monitoring in codling moth: a need for standardization     

Reyes M  Sauphanor B 《Pest management science》2008,64(9):945-953

BACKGROUND: A wide range of methods has been used for monitoring resistance in Cydia pomonella L. However, the results from specific tests are not always concordant and they generate doubts over the extent to which the data represent the field situation. Therefore, the variability in the expression of resistance mechanisms and in resistance ratios between developmental stages and their reliability in a field situation were studied. RESULTS: Neonate larvae showed the highest insecticide resistance, except to spinosad. Resistance ratios were from 2 to 9.5, depending on the insecticide and the strain tested, when compared with the susceptible strain. Spinosad exhibited the highest resistance ratio for diapausing larvae (4.4, 12.2 and 4.4 for Raz, Rdfb and RΔ strains, respectively). Enzymatic ratios were higher in adults than in neonates or diapausing larvae, and the highest enzymatic ratios were observed for mixed‐function oxidases (7.3–36.7 for adults of resistant strains). CONCLUSIONS: The sensitivity of different instars depends on the insecticides used, and there is a differential expression of resistance mechanisms between stages. For the population considered in this study, tests on neonates provided the most consistent results to the field situation, but adult response to chlorpyrifos‐ethyl and azinphos‐methyl in the laboratory was also consistent with the field results. Copyright © 2008 Society of Chemical Industry  相似文献   

Syncytial hepatitis of farmed tilapia,Oreochromis niloticus (L.): a case report   总被引：1，自引：0，他引：1  

H W Ferguson  R Kabuusu  S Beltran  E Reyes  J A Lince  J del Pozo 《Journal of fish diseases》2014,37(6):583-589

  相似文献   

Tree effects on the chemical topsoil features of oak, beech and pine forests     

Elena Marcos  Leonor Calvo  José Antonio Marcos  Ángela Taboada  Reyes Tárrega 《European Journal of Forest Research》2010,129(1):25-30

We aimed to study tree effects on the chemical properties of forest soils. We compared soil features of three types of forest ecosystems, each with four stands (replicates): beech forests (Fagus sylvatica), oak forests (dominated by Quercus pyrenaica) and pine plantations (Pinus sylvestris). Five samples from the top 10 cm of soil were taken per stand, from which pH, organic matter content (O.M.), total nitrogen (N) and available calcium (Ca²⁺), magnesium (Mg²⁺), potassium (K⁺) and sodium (Na⁺) were determined. Litter layer depth was measured at each soil sampling point. We also measured tree density and crown diameters at each stand. Our results indicated that soil samples from the four pine plantation stands were more similar while oak and beech stands were characterised by great variability in terms of soil properties and leaf litter depth. Although the identity of the dominant tree species significantly influenced several topsoil chemical properties (increase in pH and available cations in oak forests and higher organic matter and total nitrogen in beech and pine ecosystems), there were other important factors affecting soil features that may be taken under consideration. Differences between soil properties of the three types of forest ecosystems were mainly related to the characteristics of the litter layer and less related to the tree layer structure. Finally, the establishment of pine plantations in naturally deciduous tree areas made the topsoil features more homogeneous.  相似文献   

Evidence that active transmission of porcine cysticercosis occurs in Venezuela     

Cortez Alcobedes  Maria M.  Boggio  Gonzalo  Guerra  Ma. de Lourdes  de Gavidia  Magda Rodríguez  Rojas Reyes  Glenda C.  Ferrer  Elizabeth  Lares  Maria  Alviarez  Yenny  Harrison  Leslie J. S.  Parkhouse  R. Michael E. 《Tropical animal health and production》2010,42(3):531-537

There is a paucity of quantitative data on the status of porcine cysticercosis in Venezuela, information which is essential for understanding the level of disease transmission. This study was, therefore, conducted in a typical small rural community in Yaracuy State, Venezuela, where previous cases of human Taenia solium taeniasis/cysticercosis had been reported and where the free-ranging pig management practices and the lack of rudimentary sanitary facilities indicated an obvious risk for transmission of the disease. Serum samples from 52 village pigs were screened by enzyme-linked immunosorbent assays for anti-cysticercal antibodies (Ab-ELISA), using T. solium cyst fluid as the antigen and the HP10, monoclonal antibody-based, antigen trapping ELISA for parasite antigen (HP10 Ag-ELISA). Significantly, a high proportion of the animals (65.4% for the Ab-ELISA and 42.3% for the HP10 Ag-ELISA) were sero-positive. Five of the pigs, which were selected on that basis of positive tongue palpation, were killed for autopsy, and large numbers of viable cysticerci were found in the carcases. This unequivocal documentation of porcine cysticercosis in Venezuelan pigs presents clear evidence that T. solium is actively transmitted in Venezuela. Further detailed studies and implementation of appropriate control measures are therefore indicated.  相似文献   

Expression of vascular endothelial growth factor and endothelial nitric oxide synthase is increased in the placenta of sheep at high altitude in the Andes     

Parraguez VH  Atlagich MA  Urquieta B  Galleguillos M  De Los Reyes M  Kooyman DL  Araneda S  Raggi LA 《Canadian journal of veterinary research》2010,74(3):193-199

Fetal weight and the placenta of sheep at high altitude (HA) are affected by hypoxia. Placental changes (an increase in placental size and vascularization) are greater in ewes from populations that have lived for several generations at HA than in those exposed during just 1 gestation. This study investigated placental expression of vascular endothelial growth factor (VEGF) and endothelial nitric oxide synthase (eNOS), 2 molecules involved in placental angiogenesis that could be upregulated by hypoxia. Two groups of ewes were maintained at HA (3589 m) during pregnancy: HA-native ewes (group HH) and ewes native to lowlands but moved to HA immediately after the diagnosis of pregnancy (group LH). A control group (LL) was kept at sea level. Near term, placentomes were removed, weighed, and processed for immunohistochemical detection of VEGF and eNOS, as well as for vascular area measurement. Placental weight was significantly higher in the HH group than in the LH and LL groups; between the latter 2 groups there was no significant difference. The placental area occupied by vasculature was significantly greater in both the HA groups than in the LH group; the number of placentomes was greatest in the LL group. The density of VEGF and eNOS in the placentome tissue was significantly greater in both HA groups than in the LL group. Although the density of VEGF was significantly lower in the HH group than in the LH group, no differences were observed in eNOS density between the HH and LH animals. These results demonstrate that chronic hypoxia upregulates the expression of placental VEGF and eNOS, suggesting an important role of these molecules in the placental response to HA hypoxia. In addition, an attenuated response to hypoxia in VEGF synthesis may be part of the long-term process of adaptation to HA.  相似文献   

Plant water relations and edaphoclimatic conditions affecting acorn production in a holm oak (<Emphasis Type="Italic">Quercus ilex</Emphasis> L. ssp. <Emphasis Type="Italic">ballota</Emphasis>) open woodland     

F. S. Carevic  M. Fernández  R. Alejano  J. Vázquez-Piqué  R. Tapias  E. Corral  J. Domingo 《Agroforestry Systems》2010,78(3):299-308

Acorn production patterns and the annual evolution of water relations parameters of Quercus ilex ssp. ballota (Desf.) Samp. in a dehesa (an open woodland forest) subject to various soil treatments (ploughing, ploughing + sowing, control) were studied in southwest Spain from June 2006 to December 2008. The effects of soil water holding capacity and effective soil depth on soil and plant water status and acorn production were also investigated. Water parameters showed significant differences between the ploughing treatment and the control, and there were also significant temporal differences. Xylem water potential ranged from −3.4 ± 0.1 to −0.7 ± 0.2 MPa, and cuticular transpiration was 62.4–192.9 mmol H₂O kg⁻¹ s⁻¹. Acorn production did not differ significantly according to soil treatment, and showed large intra-specific variability. Individual values ranged from 0 to 1,220 g m⁻² (fresh weight). Positive relationships were found between xylem water potential, cuticular transpiration and relative water content measured at midsummer, and acorn production during the three studied years. These results suggest that climatic conditions and soil water availability have a strong influence on plant water status, and therefore on acorn development during summer. The results also reflect the ability of this species to adapt to the Mediterranean climate by preserving water during dry periods, which to a large extent can be attributed to stomatal closure at high relative water content levels, and low cuticular transpiration during these periods.  相似文献