Is sperm cryopreservation in absence of permeable cryoprotectants suitable for subfertile donkeys?     

Maria Diaz‐Jimenez  Jesús Dorado  Blasa Pereira  Cesar Consuegra  Isabel Ortiz  Manuel Hidalgo 《Reproduction in domestic animals》2019,54(Z4):102-105

Sperm from fertile donkeys have been successfully frozen in absence of permeable cryoprotectants. The aim of this study was to determine whether this cryopreservation method is suitable for subfertile donkeys in comparison to conventional sperm freezing with glycerol. Ejaculates were collected from four Andalusian Donkeys: three fertile and one subfertile. Semen was frozen with an extender containing glycerol (GLY), or adding instead sucrose 0.25 molar and 1% bovine serum albumin (SUC) as non‐permeable cryoprotectants. After thawing, samples were assessed for total (TM, %) and progressive (PM, %) sperm motility by CASA, plasma membrane integrity (PMI, %) by epifluorescence microscopy and DNA integrity (DFI, %) by SCSA. Results (mean ± SD) were compared between extenders in fertile and subfertile donkeys using the Student's t test. No differences between GLY and SUC treatments were found in the fertile group for the sperm parameters assessed. In subfertile donkey ejaculates, GLY resulted in significantly higher values than SUC for TM (25.5 ± 3.1 vs. 19.6 ± 1.9) and PM (13.3 ± 5.1 vs. 4.0 ± 1.2), respectively. In conclusion, considering all the sperm parameters assessed, sperm freezing in absence of permeable cryoprotectants may not be still an option for cryopreservation of subfertile donkey sperm.  相似文献   

Nitric oxide impacts bovine sperm capacitation in a cGMP‐dependent and cGMP‐independent manner     

Valter Luiz Maciel  Maria Clara Caldas‐Bussiere  Diego Fernando Dubeibe Marín  Carla Sobrinho Paes de Carvalho  Celia Raquel Quirino  Ana Carolina de Macedo Soares Leal 《Reproduction in domestic animals》2019,54(12):1612-1620

We aimed to elucidate whether NO acts in in vitro sperm capacitation in bovine via cGMP/PKG1 pathway. For this, cryopreserved bovine sperm were capacitated in vitro with 20 µg/ml heparin (Control) plus treatments: 1 mM L‐arginine (L‐arg, NO precursor), 50 µM Rp‐8‐Bromo‐β‐phenyl‐1,N²‐ethenoguanosine‐3′,5′‐cyclic monophosphorothioate (Rp‐8‐Br‐cGMPS, selective inhibitor of the binding site for cGMP in PKG1), 1 mM 2‐Phenyl‐4,4,5,5‐tetramethylimidazoline‐1‐oxyl 3‐oxide (PTIO, NO scavenger), and the combinations of L‐arg + RP‐8‐Br‐cGMPS and L‐arg + PTIO. Sperm motility and vigour were determined by phase‐contrast microscopy, capacitation status by chlortetracycline staining, and the intracellular concentration of cGMP was measured by ELISA. Data were subjected to analysis of variance and means compared with SNK test at 5% probability. Motility and vigour were lower in sperm treated with PTIO when compared to Control and other treatments (p < .05). The L‐arg treatment showed the highest percentage of capacitated sperm when compared to the Control and other treatments (Rp‐8‐Br‐cGMPS, L‐arg + Rp‐8‐Br‐cGMPS and PTIO) (69.8 ± 3.4%, 51.2 ± 3.0, 51.1 ± 2.1, 51.2 ± 3.0 and 45.5 ± 2.7, respectively) (p < .05). The capacitation ratio (%) was lower in treatments with Rp‐8‐Br‐cGMPS, L‐arg + Rp‐8‐Br‐cGMPS and PTIO, respectively (p < .05). Lastly, cGMP concentration (pmol/ml) was lower in PTIO and L‐arg + PTIO (1.3 ± 0.3 and 1.6 ± 0.4) and was higher in Rp‐8‐Br‐cGMPS and L‐arg + Rp‐8‐Br‐cGMPS (3.7 ± 0.4 and 4.0 ± 0.5) treatments. We showed that during in vitro capacitation of cattle: (a) NO influences sperm motility and vigour; (b) NO is associated with cGMP synthesis through two independent pathways and (c) the cGMP/PKG1 pathway has a partial role in sperm capacitation and does not involve the L‐arg/NO.  相似文献   

Linkage disequilibrium and effective population size in Gir cattle selected for yearling weight     

Alejandra M. Toro Ospina  Amanda Marchi Maiorano  Rogrio A. Curi  Guilherme L. Pereira  Maria Eugenia Zerlotti‐Mercadante  Joslaine Noely dos Santos Gonalves Cyrillo  Rusbel R. Aspilcueta‐Borquis  Josineudson A. II de V. Silva 《Reproduction in domestic animals》2019,54(12):1524-1531

Linkage disequilibrium (LD) plays an important role in genomic selection and mapping of quantitative trait loci (QTL). This study investigated the pattern of LD and effective population size (N_e) in Gir cattle selected for yearling weight. For this purpose, 173 animals with imputed genotypes (from 18 animals genotyped with the Illumina BovineHD BeadChip and 155 animals genotyped with the Bovine LDv4 panel) were analysed. The LD was evaluated at distances of 25–50 kb, 50–100 kb, 100–500 kb and 0.5–1 Mb. The N_e was estimated based on 5 past generations. The r² values (a measure of LD) were, respectively, .35, .29, .18 and .032 for the distances evaluated. The LD estimates decreased with increasing distance of SNP pairs and LD persisted up to a distance of 100 kb (r² = .29). The N_e was greater in generations 4 and 5 (24 and 30 animals, respectively) and declined drastically after the last generation (12 animals). The results showed high levels of LD and low N_e, which were probably due to the loss of genetic variability as a consequence of the structure of the Gir population studied.  相似文献   

Mycobacterium nebraskense infection in a dog in Switzerland with disseminated skin lesions     

Katrin Timm  Monika Welle  Ute Friedel  Danille Gunn‐Moore  Sophie Peterhans 《Veterinary dermatology》2019,30(3):262-e80

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Comparison of three clinical scoring systems for Culicoides hypersensitivity in a herd of Icelandic horses     

Julia E. Miller  Sabine Mann  Antonia Fettelschoss‐Gabriel  Bettina Wagner 《Veterinary dermatology》2019,30(6):536-e163

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Morphological changes of physeal cartilage and secondary ossification centres in the developing femur of the house mouse (Mus musculus): A micro‐CT based study     

Miguelangel Moncayo‐Donoso  Johana M. Guevara  Kalenia Mrquez-Flrez  Marta R. Fontanilla  Luis A. Barrera  Diego A. Garzn‐Alvarado 《Anatomia, histologia, embryologia》2019,48(2):117-124

In mammals, long bones are formed by ossification of a cartilaginous mould during early stages of development, through the formation of structures called the primary ossification centre, the secondary ossification centres (SOCs) and the physeal cartilages (PCs). The PC is responsible for long bone growth. The morphology of the PC and the SOCs varies during different stages of femoral growth. In this respect, several details involving the process of murine femoral development are lacking. In the present study, a morphological characterization of femur development from the embryonic period to adulthood in mice was studied using micro‐computed tomography (micro‐CT). To achieve this aim, femora were collected at embryonic day (E) 14.5, E16.5 and E18.5 and at postnatal day (P)1, P7, P14, P35, P46 and P52. CT images were obtained using a micro‐CT scanner (X‐SkyScan 1172; Micro Photonics) and analysed using the micro‐CT 3D visualization software Mimics (Materialise NV, Leuven, Belgium) and NRecon (Micro Photonics). The results of the present study revealed that the femur and its PCs and SOCs undergo morphological changes during different stages of development, including changes in their shape as well as position and thickness. These changes may be due to the response of the femur to mechanical loads imposed by muscle surrounding the bone during these stages of development. The result of the present study is important to improve our knowledge related to ossification and growth patterns of mouse femur during development.  相似文献   

Abnormal Sysmex XT‐2000iV DIFF scattergram in a cat with a prominent mastocytemia     

Typhaine Lavabre  Adeline Betting  Nathalie Bourgs‐Abella  Catherine Layssol‐Lamour  Catherine Trumel 《Veterinary clinical pathology / American Society for Veterinary Clinical Pathology》2019,48(4):624-629

A 2‐year‐old neutered male domestic shorthair cat was presented to the emergency service of the National Veterinary School of Toulouse (France) for acute vomiting and diarrhea with lethargy, inappetence, and adypsia for the past 48 hours. Complete blood counts were performed with the ProCyte DX at the emergency department and with the Sysmex XT‐2000iV at the laboratory 2 weeks later. The scattergrams from the two analyzers revealed similar unusual and abnormal dot plots. The Sysmex XT‐2000iV DIFF scattergram also showed no clear separation between different leukocyte populations. The eosinophil cluster was in an abnormal location compared with that of the “typical” location in a normal cat. A blood smear evaluation revealed the presence of numerous mast cells. Thus, we hypothesized that the Sysmex XT‐2000iV had detected the mast cell population, and this led to errors in the differential counts. To explore this hypothesis, we manually gated on the DIFF scattergram and performed a manual differential on the blood smear. With this new gating strategy, the Sysmex XT‐2000iV and manual differentials were similar. Thus, in the case of systemic mastocytosis, mast cells can be located between the lymphocyte, monocyte, and eosinophil clusters on scattergrams.  相似文献   

Intra‐ and inter‐rater agreement for the detection of band neutrophils and toxic change in horses     

Nicole J. Fernandez  Cornelia V. Gilroy  Catherine R. Wagg  Grace P. S. Kwong  Marie‐France Roy 《Veterinary clinical pathology / American Society for Veterinary Clinical Pathology》2019,48(4):668-676

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Oxalate‐degrading bacteria,including Oxalobacter formigenes,colonise the gastrointestinal tract of healthy koalas (Phascolarctos cinereus) and those with oxalate nephrosis     

KN Speight  M Houston‐Francis  M Mohammadi‐Dehcheshmeh  E Ebrahimie  S Saputra  DJ Trott 《Australian veterinary journal》2019,97(5):166-170

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The α2A‐adrenoceptor subtype plays a key role in the analgesic and sedative effects of xylazine     

Taisuke Kitano  Takeshi Kobayashi  Soichiro Yamaguchi  Ken‐ichi Otsuguro 《Journal of veterinary pharmacology and therapeutics》2019,42(2):243-247

Xylazine, the classical α₂‐adrenoceptor (α₂‐AR) agonist, is still used as an analgesic and sedative in veterinary medicine, despite its low potency and affinity for α₂‐ARs. Previous pharmacological studies suggested that the α_2A‐AR subtype plays a role in mediating the clinical effects of xylazine; however, these studies were hampered by the poor subtype‐selectivity of the antagonists used and a lack of knowledge of their bioavailability in vivo. Here, we attempted to elucidate the role of the α_2A‐AR subtype in mediating the clinical effects of xylazine by comparing the analgesic and sedative effects of this drug in wild‐type mice with those in α_2A‐AR functional knockout mice using the hot‐plate and open field tests, respectively. Hippocampal noradrenaline turnover in both mice was also measured to evaluate the contribution of α_2A‐AR subtype to the inhibitory effect of xylazine on presynaptic noradrenaline release. In wild‐type mice, xylazine (10 or 30 mg/kg) increased the hot‐plate latency. Furthermore, xylazine (3 or 10 mg/kg) inhibited the open field locomotor activity and decreased hippocampal noradrenaline turnover. By contrast, all of these effects were abolished in α_2A‐AR functional knockout mice. These results indicate that the α_2A‐AR subtype is mainly responsible for the clinical effects of xylazine.  相似文献