Identification of Sperm-Head Morphometric Subpopulations in Iberian Red Deer Epididymal Sperm Samples     

MC Esteso  MR Fernández-Santos  AJ Soler  V Montoro  F Martínez-Pastor  JJ Garde 《Reproduction in domestic animals》2009,44(2):206-211

Computer-assisted sperm morphometry analysis (CASMA) was used in this study to identify sperm morphometric subpopulations in Iberian red deer epididymal sperm samples. Epididymal sperm samples were collected from 37 mature stags and were divided. One portion was diluted in a Tris–citrate–egg yolk medium. A microscope slide was prepared from single extended sperm samples prior to freezing. The remainder of each sample was frozen in nitrogen vapours using a conventional protocol. After thawing, sperm smears were prepared as described for extended samples. All slides were air-dried and stained with Hemacolor^®. The sperm-head dimensions for a minimum of 145 sperm-heads were analyzed from each sample by means of the Sperm-Class Analyser^®, and the mean measurements recorded. Each sperm-head was measured for four primary sperm-head parameters, and five parameters of head shape. All sperm morphometric parameters evaluated were placed in a statistical database and a multivariate cluster analysis was performed. The clustering analyses, based on 10 867 individual spermatozoa, revealed the existence of three subpopulations (SP₁, SP₂, SP₃) of spermatozoa with different morphometric characteristics (p  <  0.001). The proportion of spermatozoa present in any of the three subpopulations remained constant (p  >  0.05) through the cryopreservation process. Pre-freeze and post-thaw sperm quality was in vitro evaluated by microscopic assessments of individual sperm motility and of plasma membrane and acrosome integrities. In conclusion, our results show that applying the CASMA techniques and multivariate cluster analyses, it was possible to determine that three subtle subpopulations of spermatozoa with different morphometric characteristics coexist in red deer semen.  相似文献   

LHRH Fusion Protein Immunization Alters Testicular Development, Ultrasonographic and Histological Appearance of Ram Testis     

H Ülker  A Y&#;lmaz  F Karaku&#;  M Yörük  C Buda&#;  DM deAvila  JJ Reeves 《Reproduction in domestic animals》2009,44(4):593-599

This study was designed to evaluate the effectiveness of a luteinizing hormone releasing hormone (LHRH) fusion protein immunization on reproductive traits in ram lambs including the changes in histology and ultrasonographic appearance of testis. Thirty native ram lambs at 19 weeks of age were divided into control (C, n = 10), immunization (I, n = 10) and castration (E, n = 10) groups. Animals in immunization group were immunized against LHRH using ovalbumin‐LHRH‐7 (OL) protein generated by recombinant DNA technology as a primary and a booster injection at 19 and 23 weeks of age respectively. Animals were bled via jugular venepuncture at 2‐week intervals to determine LHRH antibody and testosterone concentrations. Bi‐weekly ultrasonographic examination of the testes was performed to determine the changes in ultrasonographic appearance as the age increased. Biopsied testicular tissues taken at 19, 29 and 41 weeks age were also evaluated. At 41 weeks of age, animals were slaughtered. Semen and epididymis were evaluated for the presence of sperm cells. Testicular development and sperm production were suppressed in the immunized animals. Semineferous tubule diameters decreased, basal membrane of the tubule was thickened and hyalinized in immunized ram lambs. While testes of control animals gained their normal ultrasonographic appearance as the age increased, immunized animals had uniform hypoechogenic testicular structure as observed at 19 weeks of age until slaughter. Simultaneous histological and ultrasonographic evaluations indicated that the changes in testicular histology could partly be monitored via ultrasonographic imaging. Nevertheless, it is difficult to claim that ultrasonographic image reflects the exact changes in such instances. In conclusion, these results indicate that recombinant OL fusion protein is effective in immunocastration in ram lambs and has a potential to be used as an alternative to physical castration. Further research studies should be conducted to help assess reproductive status of testes from ultrasound images.  相似文献   

Sulfur nutrition effects on dinitrogen fixation of seedling alfalfa     

G. Lopez‐Jurado  D.B. Hannaway 《Journal of plant nutrition》2013,36(12):1103-1121

Greenhouse experiments with alfalfa (Medicago sativa L. cv. ‘Apollo') were performed to evaluate the effect of varied nutrient solution concentrations of S on the yield, nodulation, dinitrogen fixation, N and S concentration, and the partitioning of N and S into shoots and roots.

Sulfur treatments consisted of four levels (0, 1, 2.5, and 25 mg S/L) of added S. The experimental design was a randomized complete block, with three replications. Seeds were inoculated with commercial inoculum, planted in plastic containers of acid‐washed sand, and irrigated with nutrient solution for one minute, at 2 h intervals.

Sulfur application increased the yield of all treatments. The results demonstrated that the addition of 2.5 mg S/L to the nutrient solution, besides providing the highest total dry matter yield (12 g/72 plants), showed the highest percent yield increase (19%), acetylene reduction rate (0.426 umole ethylene/mg nodule dry wt/h), total N content (306 mg/72 plants), percent recovery of S (3.8%), and percent increase in N due to dinitrogen fixation (32%).

N:S ratios obtained were different for shoots and roots, with S application decreasing the N:S ratios. The N:S ratios of 16:1 (shoots), and 9:1 (roots) obtained in the 2.5 mg S/L treatment were found to be adequate for normal growth and development.

These data indicated that the 2.5 mg S/L treatment (2.7 mg total S/L) was optimal for alfalfa seedling development.  相似文献   

Echinococcus granulosus and other intestinal helminths: current status of prevalence and management in rural dogs of eastern Australia     

DJ Jenkins  JJ Lievaart  B Boufana  WS Lett  H Bradshaw  MT Armua‐Fernandez 《Australian veterinary journal》2014,92(8):292-298

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The efficacy of synthetic pyrethroid insecticides applied to the backline of sheep against four strains of lice (Damalinia ovis)     

PW MORCOMBE  JJ GARDNER  LE MILLAR  FC WILKINSON  GC DE  CHANEET DJ DEVEREAUX 《Australian veterinary journal》1992,69(2):35-36

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Luteolytic Effect of Prostaglandin F_2α on Bovine Corpus Luteum Depends on Cell Composition and Contact     

AJ Korzekwa  JJ Jaroszewski  I Woclawek-Potocka  MM Bah  DJ Skarzynski 《Reproduction in domestic animals》2008,43(4):464-472

Prostaglandin F_2α (PGF_2α) is a main luteolytic factor in vivo; however, its direct luteolytic influence on steroidogenic cells of bovine corpus luteum (CL) is controversial and not fully understood. The aim of the study was to clarify PGF_2α action on bovine CL in different in vivo and in vitro conditions and to examine whether the contact among all main types of CL cells is necessary for luteolytic PGF_2α action. In experiment 1, the bovine CL (day 15 of the oestrous cycle) was perfused using in vivo microdialysis system with dinoprost (an analogue of PGF_2α) for 0.5 h. Dinoprost caused a short‐time increase in progesterone (P4), whose concentration decreased thereafter (at 6‐, 10‐, 12‐ and 24‐h after treatment). In experiment 2, the direct effect of PGF_2α on P4 accumulation in CL steroidogenic cells cultured in monolayer (day 15 of the cycle) was determined. PGF_2α after 24 h of incubation increased P4 accumulation in steroidogenic CL cells. In experiment 3 steroidogenic, endothelial CL and immune cells (day 15 of the cycle) were incubated with PGF_2α in cocultures for 24 h in glass tubes and the levels of P4, stable metabolites of nitric oxide (NO) and leukotriene (LT) C₄ were determined. Although PGF_2α treatment increased P4 secretion in homogeneous steroidogenic CL cell culture, the decrease in P4 secretion in cocultures of all types of CL cells was observed. The secretion of NO and LTC₄ increased after the treatment of PGF_2α both in pure cultures of CL cells and in cocultures. The interactions between endothelial and immune cells with steroidogenic CL cells are needed for luteolytic PGF_2α action within the bovine CL. Our results indicate that the cell coculture model, including the main types of CL cells, is the most approximate to study PGF_2α role in vitro.  相似文献   

A Study on Postpartum Metritis in Iraqi Buffalo Cows: Bacterial Causes and Treatment     

OI Azawi  SN Omran  JJ Hadad 《Reproduction in domestic animals》2008,43(5):556-565

The objectives of the present study were to determine the relationship between bacteriological findings, clinical signs and histopathological changes in postpartum metritis. Evaluation of the treatment efficiency of using systemic or intra‐uterine infusion of antibiotics with some hormonal preparations for the treatment of postpartum metritis. Data were collected from 50 buffalo cows with history of calving of more than 1 month. All buffaloes were subjected to detailed clinical examination including external inspection, vaginoscopy and transrectal palpation of the cervix, uterus and ovaries. Swabs for bacteriology and biopsies for histopathology were collected from uterine lumen from each buffalo included in the present study. Bacteria identified using API systems following aerobic and anaerobic cultures. Vaginal mucus scored for character, odour and estimation of polymorphonuclear cells (PMNs). Treatment conducted using oxytetracycline in local intrauterine infusion or systemically with hormonal treatment including prostaglandinF2α (PGF2α) and oestradiol benzoate. Results revealed that the most predisposing factor for postpartum uterine infection was retained placenta and toxic puerperal metritis. The most prevalent bacteria in uterine lumen were Escherichia coli, Archanobacterium pyogenes, Bacteroides fragilis and Fusobacterium necrophorum the most prevalent bacteria in buffaloes with postpartum metritis. A. pyogenes and F. necrophorum were an important pathogens causing severe uterine inflammation as found in histopathological examinations. Buffaloes with postpartum metritis showed good clinical cure when oxytetracycline injected systemically with PGF2α. Intrauterine infusion of oxytetracycline had no advantage for the treatment of uterine infection in buffalo cows with postpartum metritis. PGF2α improved clinical cure of buffaloes with postpartum metritis.  相似文献   

Genetic parameter estimation and implementation of the genetic evaluation for gaits in a breeding program for assisted-therapy in donkeys     

Francisco Javier Navas González  Jordi Jordana Vidal  Jose Manuel León Jurado  Amy Katherine McLean  Gabriela Pizarro Inostroza  Juan Vicente Delgado Bermejo 《Veterinary research communications》2018,42(2):101-110

Genetic analyses in donkeys are likely to face compromises in terms of sample size and population structure. This study aims at implementing a suitable model to estimate breeding values and genetic parameters for gaits in Andalusian donkeys. Empirical observation revealed that ambling donkeys (showing a slightly uneven, non-isochronous 1–2, 3–4 lateral sequence gait) did not walk (i.e. presented an isochronous, even 1-2-3-4 sequence gait) and vice versa. However, the two donkey groups could trot, equally. In this study, 2700 gait records were registered from 300 donkeys. The sample included 1350 gait records from 169 ambling/trotting donkeys and 1350 gait records from 131 walking/trotting donkeys. Fixed effects included year, season, sex, farm/owner, husbandry system, weather, ground type and appraisers. Weight and age were included as covariates. MTDFREML software was used to estimate (co)variance components, genetic parameters and predict breeding values and their accuracies in both sets, separately. Gaits’ heritability?±?SE estimates were 0.56?±?0.155, 0.53?±?0.317 and 0.67?±?0.166 for amble, walk and trot, respectively. Genetic correlations were 0.31?±?0.216, 0.42?±?0.115 and 0.28?±?0.178, for amble and walk, amble and trot and walk and trot, respectively. Not all gaits are suitable to treat every human sensomotor condition. We developed a locomotion selection index, assessing the relative loss/gain in index accuracy when each gait modality was excluded to develop different gait specific therapeutic lines to genetically select the best performing donkeys from each gait modality. Our results suggest that gait genetic lines could be developed and may be potential selection criteria to consider in assisted-therapy donkey breeding programs.  相似文献   

An In Vitro Evaluation of Biochemical Processes Involved in Lead‐Induced Changes on Ram Spermatozoa          下载免费PDF全文

P Castellanos  A Maroto‐Morales  O García‐Álvarez  JJ Garde  R Mateo 《Reproduction in domestic animals》2016,51(3):421-427

Lead (Pb²⁺) is a toxic heavy metal which interferes with several physiological processes regulated by Ca²⁺, including those characterized by changes of the membrane stability and the motility of spermatozoa necessary for the fertilization of the oocyte. In this study, ejaculated sperm from six rams (Ovis aries) have been incubated in vitro with or without 50 ng Pb²⁺/ml during 30 min and in the presence or absence of three different potential modulators of the effects of Pb²⁺ on changes in the sperm membrane before fertilization: charybdotoxin, quinacrine and staurosporine. Sperm samples incubated with Pb²⁺ have shown significant reductions in acrosome integrity and sperm viability and an increase in progressive movement. None of the studied potential modulators had a protective effect against Pb²⁺ action. On the contrary, Pb²⁺‐incubated sperm in the presence of staurosporine had lower acrosome integrity, and lower sperm viability was observed when spermatozoa were incubated with Pb²⁺ + charybdotoxin. Quinacrine was the only tested substance capable of increasing the concentration of Pb²⁺ in spermatozoa; thus, the enhancement of Pb²⁺ effects produced by staurosporine and charybdotoxin was not produced by an increased uptake of Pb²⁺ by spermatozoa. However, the increase of intracellular Pb²⁺ in those spermatozoa incubated with quinacrine did not result in an adverse effect on sperm motility or viability although the acrosome integrity was negatively affected.  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