Effect of permeable cryoprotectant‐free vitrification on DNA fragmentation of equine oocyte–cumulus cells     

Blasa Carmen Pereira  Isabel Ortiz  Jesús Manuel Dorado  Maria Angeles Diaz‐Jimenez  Cesar Consuegra  Jaime Gosalvez  Manuel Hidalgo 《Reproduction in domestic animals》2019,54(Z3):53-56

DNA fragmentation of cumulus cells could be used as an indicator of oocyte vitrification success as an indirect indicator of the quality of the oocyte. This study was designed to compare the DNA fragmentation of post‐mortem equine cumulus cells before or after vitrification in the absence of permeable cryoprotectant agents. Cumulus–oocyte complexes (COCs; n = 56) were recovered from slaughterhouse ovaries and subjected to in vitro maturation (42 hr/38.2°C/5%CO₂) before (control group) or after a permeable cryoprotectant‐free vitrification method using 1 M sucrose (vitrification group). After in vitro maturation, COCs were denuded, and cumulus cells were washed and stored at ?80°C until thawing. Cumulus cell samples were processed with the chromatin dispersion test (Ovoselect, Halotech DNA, Spain). Low, high and total DNA fragmentation percentages of cumulus cells were recorded and compared between the two groups by Student's t test. Results were expressed as mean ± SEM. The vitrified group resulted in significantly higher (p < 0.05) percentages for low (16.81 ± 1.62 vs. 6.63 ± 0.77) and total (21.14 ± 1.84 vs. 12.76 ± 1.48) DNA fragmentation of cumulus cells. There were no significant differences between groups for high DNA fragmentation of cumulus cells. In conclusion, permeable cryoprotectant‐free vitrification of equine oocytes increased the total DNA fragmentation rate of cumulus cells but protected them against high DNA fragmentation rates. Further studies are needed to examine the relationship between DNA fragmentation of cumulus cells and the developmental competence of equine oocytes.  相似文献   

The Effect of Oxidative Stress on Thawed Bulk‐Sorted Red Deer Sperm          下载免费PDF全文

L Anel‐López  O García‐Álvarez  I Parrilla  D Del Olmo  MR Fernández‐Santos  AJ Soler  A Maroto‐Morales  JA Ortiz  DV Alkmin  T Tarantini  J Roca  EA Martínez  JM Vazquez  JJ Garde 《Reproduction in domestic animals》2016,51(3):407-414

The aims of this study were to assess the effects of the sex‐sorting process on post‐thaw sperm quality as well as on induced oxidative stress damage (H₂O₂ 0 mm  = H000; H₂O₂ 50 mm  = H050; H₂O₂ 100 mm  = H100) and the protective action of reduced glutathione (GSH) and Trolox, when comparing sorted (BSS) and non‐sorted (NS) red deer spermatozoa incubated at 37°C. Sperm samples from three stags were collected by electroejaculation and frozen. Immediately after thawing, sperm motility was higher (p < 0.05) for NS (59% ± 3.3) than BSS (36.9% ± 5.8) sperm. Furthermore, the percentage of apoptotic sperm was higher (p < 0.05) for BSS (21.6% ± 5.0) than NS sperm (14.6% ± 1.2). The presence of H₂O₂ increased DNA damage in NS (H000 = 4.1% ± 0.9; H050 = 9.3% ± 0.7; and H100 = 10.9% ± 2.3), but not in BSS sperm. However, in the presence of oxidant, GSH addition improved (p < 0.05) sperm motility in both groups of sperm samples as compared to their controls (NS: 44.5 ± 4.8 vs 21.1 ± 3.9 and BSS: 33.3 ± 8.1 vs 8.9 ± 1.8). These results demonstrate that the sperm‐sorting process induces sublethal effects, albeit selecting a sperm population with a chromatin more resistant to oxidative stress than that in non‐sorted sperm. Moreover, addition of GSH at 1 mm may be a good choice for maintaining the quality of stressed sperm samples, unlike Trolox, which inhibited sperm motility.  相似文献   

First report of coconut lethal yellowing disease in Guatemala     

F. Mejía  M. Palmieri  C. Oropeza  M. Doyle  N. Harrison  E. Aguilar  M. Narváez  R. Estrada  G. Ortiz 《Plant pathology》2004,53(6):800-800

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Nontarget-site resistance due to rapid physiological response in 2,4-D resistant Conyza sumatrensis: reduced 2,4-D translocation and auxin-induced gene expression     

Amanda dos Santos Souza  Jessica Ferreira Lourenço Leal  Jacob Samuel Montgomery  Mirella Farinelli Ortiz  André Lucas Simões Araujo  Sarah Morran  Marcelo Rodrigues Alves de Figueiredo  Ana Claudia Langaro  Luiz Henrique Saes Zobiole  Scott Jay Nissen  Todd Adam Gaines  Camila Ferreira de Pinho 《Pest management science》2023,79(10):3581-3592

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Combining the effects of supplementary feeding and copper oxide needles for the control of gastrointestinal nematodes in browsing goats     

Martínez Ortiz de Montellano C  Vargas-Magaña JJ  Aguilar-Caballero AJ  Sandoval-Castro CA  Cob-Galera L  May-Martínez M  Miranda-Soberanis R  Hoste H  Cámara Sarmiento R  Torres-Acosta JF 《Veterinary parasitology》2007,146(1-2):66-76

The aim was to assess the benefits obtained from combining supplementary feeding and copper needles (COWP), compared to the use of both approaches independently, for the control of gastrointestinal nematode (GIN) infections in browsing kids. Forty-four nematode free Criollo kids were exposed to natural parasite infection. The kids were divided into six experimental groups: not treated, supplemented (NT-S), not treated, not supplemented (NT-NS), moxidectin treated, supplemented (M-S), moxidectin treated not supplemented (M-NS), copper treated, supplemented (COWP-S) and copper treated, non-supplemented (COWP-NS). Copper treated groups received Copinox (2 g capsules) on day 0 and on day 60 of the trial. Moxidectin treated groups received Cydectin (0.2 mg/kg of body weight s.c.) every 28 days. Three of the groups received individual supplementation (100 g of feed/day fresh basis; 74% sorghum: 26% soybean meal; NT-S, M-S and COWP-S) and the other three groups were not supplemented (NT-NS, M-NS and COWP-NS). Animals browsed native vegetation (6.5 h/day) during the wet season (154 days). Kids were weighed every 14 days to determine live weight gain (LWG) and blood and faecal samples were obtained to determine packed cell volume (PCV), haemoglobin (Hb), peripheral eosinophil counts (PEC) and faecal egg counts (FEC). At the end of the trial, four kids of each group were euthanatized (six kids in each COWP treated group). Worm burdens, female worm lengths and prolificacy were determined. Liver samples were used to determine copper concentration and were stained with haematoxylin-eosin to determine microscopic lesions. Animals receiving the combination of supplementary feeding and COWP improved their LWG, PCV and Hb to similar levels of animals with suppressive AH treatment. This was not the case when COWP was used without supplementation. Liver copper concentration in COWP treated groups increased significantly especially in the COWP-NS kids but this was not associated with liver lesions or clinical signs. Post-mortem Haemonchus contortus and Trichostrongylus colubriformis worm counts had a tendency to be reduced in the different groups (66-35% reduction) compared to NT-NS group at the end of the trial (P>0.05). Also, COWP treatment and/or supplementation reduced female worm length of T. colubriformis and prolificacy of H. contortus and T. colubriformis. This study, confirmed the value of nutritional supplementation in the control of GIN in growing kids. The use of COWP in addition to supplementation had a limited contribution on the kids' resilience against GIN. This may be due to the reduced infection of H. contortus during this trial.  相似文献   

Genotype × environment interaction and selection for drought adaptation in sweetpotato (Ipomoea batatas [L.] Lam.) in Mozambique     

Maria I. Andrade  Abdul Naico  José Ricardo  Raúl Eyzaguirre  Godwill S. Makunde  Rodomiro Ortiz  Wolfgang J. Grüneberg 《Euphytica》2016,209(1):261-280

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Quantitative immunoenzymatic detection of viral encephalopathy and retinopathy virus (betanodavirus) in sea bass Dicentrarchus labrax          下载免费PDF全文

N Nuñez‐Ortiz  V Stocchi  A Toffan  F Pascoli  N Sood  F Buonocore  S Picchietti  C Papeschi  A R Taddei  K D Thompson  G Scapigliati 《Journal of fish diseases》2016,39(7):821-831

Viral encephalopathy and retinopathy disease caused by betanodavirus, genus of the family Nodaviridae, affects marine, wild and farmed species including sea bass, one of the most important farmed species in Europe. This work describes a reliable and sensitive indirect ELISA assay to detect betanodavirus in biological samples using a polyclonal antiserum (pAb 283) against the 283/I09 virus strain, the most common red‐spotted grouper nervous necrosis virus (RGNNV) genotype in the Mediterranean area, and a capture‐based ELISA using a monoclonal antibody (mAb 4C3) specific to a common epitope present on the capsid protein. Using adsorbed, purified VERv preparation, the detection limit of indirect ELISA was 2 μg mL^?1 (3 × 10⁵ TCID50 per mL), whereas for capture‐based ELISA, the sensitivity for the antigen in solution was 17 μg mL^?1 (35 × 10⁵ TCID50 per mL). The capture‐based ELISA was employed to detect VERv in brain homogenates of in vivo infected sea bass and resulted positive in 22 of 32 samples, some of these with a high viral load estimates (about 1.1 × 10⁸ TCID50 per mL). The ELISA system we propose may be helpful in investigations where coupling of viral content in fish tissues with the presence of circulating VERv‐specific IgM is required, or for use in samples where PCR is difficult to perform.  相似文献   

The effect of dietary oxidized lipid levels on growth performance,antioxidant enzyme activities,intestinal lipid deposition and skeletogenesis in Senegalese sole (Solea senegalensis) larvae     

A. Boglino  M.J. Darias  A. Estévez  K.B. Andree  C. Sarasquete  J.B. Ortiz‐Delgado  M. Solé  E. Gisbert 《Aquaculture Nutrition》2014,20(6):692-711

Fish tissues, particularly rich in n‐3 PUFA, are prone to lipid peroxidation that can damage cellular membranes, cause severe lesions and subsequently incidences of disease and mortality. However, fish possess antioxidant defences, such as vitamin E (VE) and antioxidant enzymes, to protect them against oxidative damage. This study investigated the effects of an increasing gradient of oxidized dietary lipid on the survival, growth performance, skeletogenesis and antioxidant defensive processes occurring in Senegalese sole (Solea senegalensis) larvae. Four groups of fish were fed live prey enriched with experimental emulsions containing an increasing gradient of oxidized oil: non‐oxidized, NO+VE, 34.5 nmol MDA g^?1 w.w.; mildly oxidized, MO+VE, 43.1 nmol MDA g^?1 w.w.; highly oxidized, HO+VE, 63.3 nmol MDA g^?1 w.w. and highly oxidized without VE, HO‐VE, 78.8 nmol MDA g^?1 w.w. The oxidation levels increased in enriched rotifers following the oxidation gradient of the emulsions, but were not affected in enriched Artemia metanauplii. The oxidation status of Senegalese sole larvae increased during development, but this was not related to the dietary treatments. The increasing dietary oxidation levels did not affect the fatty acid profile, survival, growth performance and metamorphosis processes of sole larvae. Senegalese sole seem to activate antioxidant defence mechanisms in response to the increasing amounts of dietary peroxidized lipids, in a manner efficiently enough to prevent detection of any alterations of these physiological processes. Antioxidant systems and detoxification mechanisms appeared to occur through the consumption of dietary α‐tocopherol, the activation of the antioxidant enzymes (catalase, superoxide dismutase, glutathione S‐transferase, glutathione reductase) and the retention of oxidized fat in the intestinal enterocytes for detoxification prior to their utilization. However, fish fed the highest oxidized diet presented a reduction in bone mineralization, but lower incidence of deformities in the vertebral and caudal regions than fish fed the other diets. This study exemplifies the importance of rearing Senegalese sole larvae on non‐oxidized diets during the early larval development to avoid detrimental consequences in older fish, most notably in the process of skeletogenesis.  相似文献   

Transmission of lymphocystis disease virus to cultured gilthead seabream,Sparus aurata L., larvae     

I Cano  E J Valverde  E Garcia‐Rosado  M C Alonso  B Lopez‐Jimena  J B Ortiz‐Delgado  J J Borrego  C Sarasquete  D Castro 《Journal of fish diseases》2013,36(6):569-576

The transmission of lymphocystis disease virus (LCDV) to gilthead seabream, Sparus aurata L., larvae was investigated using fertilized eggs from a farm with previous reports of lymphocystis disease. LCDV genome was detected by PCR‐hybridization in blood samples from 17.5% of the asymptomatic gilthead seabream broodstock analysed. Using the same methodology, eggs spawned from these animals were LCDV positive, as well as larvae hatched from them. The presence of infective viral particles was confirmed by cytopathic effects development on SAF‐1 cells. Whole‐mount in situ hybridization (ISH) and immunohistochemistry (IHC) showed the presence of LCDV in the epidermis of larvae hatched from LCDV‐positive eggs. When fertilized eggs were disinfected with iodine, no viral DNA was detected either in eggs (analysed by PCR‐hybridization) or in larvae (PCR‐hybridization and ISH). These results suggest the vertical transmission of LCDV, the virus being transmitted on the egg surface. Larvae hatched from disinfected eggs remain LCDV negative during the endotrophic phase, as showed by PCR‐hybridization, ISH and IHC. After feeding on LCDV‐positive rotifers, viral antigens were observed in the digestive tract, which suggests that viral entry could be achieved via the alimentary canal, and that rotifers can act as a vector in LCDV transmission to gilthead seabream larvae.  相似文献   

Antiviral activity of casein and αs2 casein hydrolysates against the infectious haematopoietic necrosis virus,a rhabdovirus from salmonid fish     

S Rodríguez Saint‐Jean  A De las Heras  W Carrillo  I Recio  J B Ortiz‐Delgado  M Ramos  J A Gomez‐Ruiz  C Sarasquete  S I Pérez‐Prieto 《Journal of fish diseases》2013,36(5):467-481

Salmonid fish viruses, such as infectious haematopoietic necrosis virus (IHNV), are responsible for serious losses in the rainbow trout and salmon‐farming industries, and they have been the subject of intense research in the field of aquaculture. Thus, the aim of this work is to study the antiviral effect of milk‐derived proteins as bovine caseins or casein‐derived peptides at different stages during the course of IHNV infection. The results indicate that the 3‐h fraction of casein and α_S2‐casein hydrolysates reduced the yield of infectious IHNV in a dose‐dependent manner and impaired the production of IHNV‐specific antigens. Hydrolysates of total casein and α_S2‐casein target the initial and later stages of viral infection, as demonstrated by the reduction in the infective titre observed throughout multiple stages and cycles. In vivo, more than 50% protection was observed in the casein‐treated fish, and the kidney sections exhibited none of the histopathological characteristics of IHNV infection. The active fractions from casein were identified, as well as one of the individual IHNV‐inhibiting peptides. Further studies will be required to determine which other peptides possess this activity. These findings provide a basis for future investigations on the efficacy of these compounds in treating other viral diseases in farmed fish and to elucidate the underlying molecular mechanisms of action. However, the present results provide convincing evidence in support of a role for several milk casein fractions as suitable candidates to prevent and treat some fish viral infections.  相似文献