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F Minervini R Guastamacchia F Pizzi ME Dell’Aquila VL Barile 《Reproduction in domestic animals》2013,48(2):317-324
Flow cytometry is a useful tool that provides an accurate, objective and rapid evaluation of semen quality. The use of this technique could significantly improve the quality of buffalo semen samples used in artificial insemination. This study was carried out to evaluate, by flow cytometry, frozen–thawed buffalo spermatozoa quality parameters such as sperm viability by SYBR‐14/propidium iodide staining; mitochondrial function by JC‐1 potentiometric probe; sperm chromatin stability (SCSA) by acridine orange; and acrosome reaction (AR) by FITC‐PNA staining. Semen samples from five Italian Mediterranean buffalo bulls were used. Sperm viability was not different between bulls and ranged from 33.4% to 43.6%. A consistent rate (55.1 ± 10.8%) of sperm cells showed high mitochondrial membrane potential (Δψhigh), with no significant differences between subjects. Sperm chromatin structure assay differed significantly between the five buffalo bulls; moreover, data showed high stability within each buffalo. DNA fragmentation indexes (DFI), such as %‐DFI, ‐DFI, SD‐DFI, were 11.2 ± 8.6, 153.3 ± 24.6 and 81.6 ± 21.2, respectively. Regarding AR, the percentage of acrosome‐reacted live (ARL) and acrosome‐reacted dead (ARD) spermatozoa was 0.3 ± 0.2 and 15.3 ± 5.5, respectively. This functional parameter differed significantly between buffalo bulls and showed high stability. Following to Ca2+ ionophore A23187 for 3 h, AR significantly differed between subjects and was characterized by an increase in both ARL (10.8%) and ARD population (22.0%). This study indicates that flow cytometry could be a useful tool for a quick multiparametric evaluation of sperm quality in buffalo. In particular, SCSA and AR resulted in sperm functional parameters sensitive enough for the diagnosis of frozen‐thawed semen fertilizing potential. 相似文献
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Long‐term Effects of Pyrethrin and Cyfluthrin,a Type II Synthetic Pyrethroid,Insecticide Applications on Bull Reproductive Parameters 
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下载免费PDF全文 JL Stewart CF Shipley FA Ireland VL Jarrell CL Timlin DW Shike TL Felix 《Reproduction in domestic animals》2016,51(5):680-687
The objectives of this study were to determine effects of cyfluthrin and pyrethrin spray products, used in combination with cyfluthrin topical and ear tag applications, on bull reproductive parameters over 18 weeks. Angus or Angus x Simmental bulls were randomly assigned to one of three treatment groups: (i) no exposure to pyrethrins/cyfluthrin (CONT; n = 10), (ii) cyfluthrin ear tag and topical applications (ET; n = 10), or (iii) cyfluthrin ear tag, topical, premise spray and pyrethrin fog spray applications (ET+S; n = 8). Bull body weight was measured every 3 week, and body condition score and scrotal circumference were recorded on weeks 0, 9 and 18. Semen and serum were collected every 3 weeks for sperm evaluation and testosterone measurement, respectively. There was a treatment × week interaction (p < 0.01) for sperm with primary defects; bulls in CONT group had a greater (p = 0.01) percentage of sperm with primary defects than bulls treated with insecticides at week 18. Overall and progressive sperm motility, normal sperm morphology, secondary sperm defects and serum testosterone concentrations changed (p < 0.01) over time in all bulls; however, treatment did not affect (p ≥ 0.13) any of these parameters. There were also no treatment effects (p ≥ 0.08) on bull body weight, body condition score or scrotal circumference. The use of pyrethrin‐ and cyfluthrin‐based insecticides, regardless of application, did not negatively affect reproductive parameters in beef bulls when administered over 18 weeks. 相似文献
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EV Abakushina Y Morita Y Kaedei F Tanihara Z Namula VL Viet T Otoi 《Reproduction in domestic animals》2011,46(3):423-427
Culture techniques of antral follicle–like structure (AFLS) derived from cumulus–oocyte complexes (COCs) might provide important insights into follicular development and oocyte maturation. This study was undertaken to investigate the effects of embedding bovine COCs individually (one COC) or in groups (4–5 COCs) in collagen gels on the formation of AFLS and the meiotic status of oocytes. The observations of AFLS formation were performed every second day for 14 days. The AFLS was formed at Day 2 or 4 after the start of culture (Day = 0), irrespective of the culture methods. The mean diameters of AFLS during Days 4–14 using the individual culture method were significantly higher (p < 0.05) than those using the group culture method. However, the AFLS formation rate in the individual culture method was significantly lower compared to that in the group culture method (26.1% vs 62.7%, p < 0.01). Almost all oocytes had undergone the germinal vesicle breakdown stage, irrespective of the culture method or AFLS formation. In conclusion, comparison with the individual culture method revealed that the mean diameters of AFLS in the group culture method were smaller, but more COCs formed AFLS. The group culture method might be useful for evaluating the various hypotheses of follicular formation and interfollicular communication. However, improvement of the group culture system is necessary to prevent the meiotic resumption of oocytes, because the AFLS formation is dependent on the cumulus/granulosa cells surrounding oocytes. 相似文献
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Hidalgo-Cuadrado N Pérez-Galende P Manzano T De Maria CG Shnyrov VL Roig MG 《Journal of agricultural and food chemistry》2012,60(19):4765-4772
Aqueous crude extracts of a series of plant wastes (agricultural, wild plants, residues from sports activities (grass), ornamental residues (gardens)) from 17 different plant species representative of the typical biodiversity of the Iberian peninsula were investigated as new sources of peroxidases (EC 1.11.1.7). Of these, lentil (Lens culinaris L.) stubble crude extract was seen to provide one of the highest specific peroxidase activities, catalyzing the oxidation of guaiacol in the presence of hydrogen peroxide to tetraguaiacol, and was used for further studies. For the optimum extraction conditions found, the peroxidase activity in this crude extract (110 U mL(-1)) did not vary for at least 15 months when stored at 4 °C (k(inact) = 0.146 year(-1), t(1/2 inact) = 4.75 year), whereas, for comparative purposes, the peroxidase activity (60 U mL(-1)) of horseradish (Armoracia rusticana L.) root crude extract, obtained and stored under the same conditions, showed much faster inactivation kinetics (k(inact) = 2.2 × 10(-3) day(-1), t(1/2 inact) = 315 days). Using guaiacol as an H donor and a universal buffer (see above), all crude extract samples exhibited the highest peroxidase activity in the pH range between 4 and 7. Once semipurified by passing the crude extract through hydrophobic chromatography on phenyl-Sepharose CL-4B, the novel peroxidase (LSP) was characterized as having a purity number (RZ) of 2.5 and three SDS-PAGE electrophoretic bands corresponding to molecular masses of 52, 35, and 18 kDa. The steady-state kinetic study carried out on the H(2)O(2)-mediated oxidation of guaiacol by the catalytic action of this partially purified peroxidase pointed to apparent Michaelian kinetic behavior (K(m)(appH(2)O(2)) = 1.87 mM; V(max)(appH(2)O(2)) = 6.4 mM min(-1); K(m)(app guaicol) = 32 mM; V(max)(app guaicol) = 9.1 mM min(-1)), compatible with the two-substrate ping-pong mechanism generally accepted for peroxidases. Finally, after the effectiveness of the crude extracts of LSP in oxidizing and removing from solution a series of last-generation dyes present in effluents from textile industries (1) had been checked, a steady-state kinetic study of the H(2)O(2)-mediated oxidation and decolorization of Green Domalan BL by the catalytic action of the lentil stubble extract was carried out, with the observation of the same apparent Michaelian kinetic behavior (K(m)(appGD) = 471 μM; V(max)(appGD)= 23 μM min(-1)). Further studies are currently under way to address the application of this LSP crude extract for the clinical and biochemical analysis of biomarkers. 相似文献
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Oestrogen and Progesterone Receptors and COX‐2 Expression in Endometrial Biopsy Samples During Maternal Recognition of Pregnancy in Llamas (Lama glama) 下载免费PDF全文
下载免费PDF全文 CP Bianchi A Meikle MA Benavente MA Álvarez VL Trasorras MH Miragaya E Rodríguez MA Aba 《Reproduction in domestic animals》2015,50(6):980-988
Endometrial expression of oestrogen receptor‐α (ERα), progesterone receptor (PR) and cyclooxigenase‐2 (COX‐2) was evaluated in non‐pregnant and pregnant llamas during the period when luteolysis/maternal recognition of pregnancy is expected to occur. Females (n = 28) were divided into two groups: non‐pregnant llamas were induced to ovulate with a Buserelin injection, and endometrial biopsies were obtained on day 8 (n = 5) or 12 (n = 5) post‐induction of ovulation. Animals of the pregnant group (n = 18) were mated with a fertile male. Pregnancy was confirmed by the visualization of the embryo collected by transcervical flushing in 5 of 9 animals on day 8 post‐mating and by progesterone profile on day 12 post‐mating in 4 of 9 animals, when endometrial biopsies were obtained. An immunohistochemical technique was used to evaluate receptors population and COX‐2 expression. Pregnant llamas showed a higher percentage of positive cells and stronger intensity for ERα than for non‐pregnant llamas in stroma on day 8 and in the luminal epithelium on day 12 post‐induction of ovulation, while a deep decrease in endometrial PR population was reported in pregnant llamas on that day in luminal and glandular epithelia and stroma. In the luminal epithelium, COX‐2 expression was lower in pregnant than in non‐pregnant animals. Briefly, the increase of ERα in pregnant llamas gives further support to the hypothesis that oestrogens are involved in the mechanism of maternal recognition of pregnancy. Endometrial PR decrease in pregnant llamas might be a necessary event to allow the expression of proteins involved in conceptus attachment, a mechanism widely accepted in other species. Moreover, embryo seems to attenuate maternal PGF2α secretion during early pregnancy by decreasing the endometrial expression of COX‐2 in the luminal epithelium of pregnant llamas. 相似文献
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VL Grillo KE Arzey PM Hansbro AC Hurt S Warner J Bergfeld GW Burgess B Cookson CJ Dickason M Ferenczi T Hollingsworth MDA Hoque RB Jackson M Klaassen PD Kirkland NY Kung S Lisovski MA O'Dea K O'Riley D Roshier LF Skerratt JP Tracey X Wang R Woods L Post 《Australian veterinary journal》2015,93(11):387-393
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VL Trasorras MG Chaves MH Miragaya M Pinto B Rutter M Flores A Agüero 《Reproduction in domestic animals》2009,44(3):359-364
The aim of the present study was two fold. Experiment I: evaluate the effect of buserelin on llama's oocyte maturation after exogenous follicular activity suppression, followed by ovarian superstimulation with different doses of equine chorionic gonadotropin (eCG). Experiment II: compare the number of follicles aspirated and the number of cumulus–oocyte complexes (COCs) recovered according to different doses of eCG followed by buserelin. Experiment I consisted in a control group (without buserelin) and a treatment group (with buserelin), both subdivided according to eCG dose administered: A: 500 IU; B: 1000 IU; C: 1500 IU. The treatment group received a single i.v. dose of 8 μg of buserelin when two or more dominant follicles were found at ultrasound evaluation and 20 h later were subjected to surgery. In group A, 83% of the llamas did not respond to superstimulation. In groups B and C differences were observed between the control and the treatment groups for the degree of COCs maturation (p < 0.05). In experiment II animals were divided into two groups according to the eCG dose administered: 1000 and 1500 IU. Twenty hours before surgery females received a single i.v. dose of 8 μg of buserelin. Average number of follicles aspirated and COCs recovered was higher (p < 0.05) with the administration of 1500 IU of eCG. A larger number of expanded COCs were obtained from follicles ≥7 mm in diameter. We conclude that buserelin aids the recovery of a larger number of expanded COCs. Administration of 1500 IU of eCG produces a higher number of follicles for aspiration and number of COCs recovered. 相似文献