Effects of Prolonged in vitro Culture and Cryopreservation on Viability,DNA Fragmentation,Chromosome Stability and Ultrastructure of Bovine Cells from Amniotic Fluid and Umbilical Cord     

ER da Cunha  CF Martins  CG Silva  HC Bessler  SN Báo 《Reproduction in domestic animals》2014,49(5):806-812

The objective of this work was to study cellular types that did not participated in the gastrulation process, amniotic fluid cells (AFCs) and umbilical cord cells (UCCs), in conditions of long‐term culture and cryopreserved with different solutions. The AFCs and UCCs were used in a comparative study with ear fibroblast cells (EFCs) that were cultured in vitro until 20 cellular passages and cryopreserved in 10% dimethylsulphoxide (DMSO), 5% dimethyl formamide (DMF) and 7% glycerol (Gly) solutions. The cellular viability, ultrastructure, DNA fragmentation and chromosome stability were evaluated to determine the cellular type most resistant. In all cell types, it was possible to evaluate the AFCs until 15 passages and UCCs until 20 passages with different periods of cellular growth to reach the confluence phase. Solutions containing 10% DMSO ensured viability of 90.33 ± 5.58%, 90.56 ± 4.40% and 81.90 ± 3.31%, respectively for EFCs, AFCs and UCCs, being significantly more efficient and with less variation than other cryoprotectant solutions. The AFCs were more sensitive to cryopreservation and presented low viability rate at the passage 20 (17.2 ± 8.87%). There was no change in karyotype and nuclear fragmentation was low in all cellular passages studied. With the scanning electron analysis was possible the characterization of AFCs and UCCs in suspension. The three cellular types of cells presented different shapes and characteristics on the surface. The results demonstrate that bovine AFCs and UCCs can be isolated, cultured in vitro and cryopreserved in 10% DMSO, not causing damage to DNA and chromosomes. The UCCs were more resistant than AFCs in all aspects.  相似文献   

Effect of oxytocin on suppression of oestrus in mares exhibiting normal oestrous cycles     

EK Gee  L Gillespie  CF Bolwell 《New Zealand veterinary journal》2013,61(3):189-193

Abstract

AIM: To compare the efficacy of oxytocin given once daily, either I/V or I/M, on Days 7–14 post-ovulation, on the expression of oestrus in mares through to 65 days post-ovulation.

METHODS: Eighteen mares of various breeds that were displaying normal oestrous cycles were randomly assigned to one of three treatment groups on the day of ovulation (Day 0), detected using transrectal ultrasonography. Mares in the control group (n = 6) were given 1 mL saline I/V; mares in the I/V and I/M groups (n = 6 per group) were injected with 10 IU oxytocin I/V and I/M, respectively. All treatments were given once daily on Days 7–14. Mares were teased by a stallion three times per week, up to 65 days post-ovulation, to detect oestrous or dioestrous behaviour. Ovarian follicular and luteal activity were monitored using transrectal ultrasonography three times weekly, and daily when a follicle >30 mm diameter was present until ovulation. Blood samples were collected weekly for analysis of concentrations of progesterone in serum. Prolonged dioestrus was defined as a period of >30 days of dioestrous behaviour after Day 0, confirmed by detection of corpora lutea and concentrations of progesterone in serum >4 nmol/L.

RESULTS: Overall, 8/18 (44%) mares showed prolonged dioestrus. These included 2/6 (33%) mares in the control group, compared with 5/6 (83%) and 1/6 (16%) mares in the I/V and I/M groups, respectively (p = 0.11). The median duration of the first dioestrus was longer for the I/V group (64 (min 16, max 66) days) compared with the control group (18 (min 12, max 64) days) (p = 0.05), but was not different between the control group and the I/M group (16 (min 13, max 65) days) (p = 0.57). For all mares there was strong agreement between teasing behaviours, ultrasonographic assessment of ovarian activity, and concentration of progesterone in serum.

CONCLUSIONS AND CLINICAL RELEVANCE: This study found that low doses of oxytocin did not increase the proportion of mares with prolonged dioestrus, compared with controls, although I/V oxytocin did increase the median duration of dioestrus. The results must be interpreted with some caution as group numbers were small, and a variety of breeds were used. Further investigation of oxytocin given I/V may be warranted as a potential method of oestrus suppression in mares exhibiting oestrous cycles that is low cost, safe and well-tolerated, and potentially reversible with prostaglandin.  相似文献   

Precision of a Novel Computed Tomographic Method for Quantification of Femoral Varus in Dogs and an Assessment of the Effect of Femoral Malpositioning     

Bill Oxley MA VetMB CertSAS  Toby J. Gemmill BVSc MVM DSAS Diplomate ECVS  Jonathan Pink BSc BVetMed CertSAS Diplomate ECVS  Stephen Clarke BVM&S DSAS Diplomate ECVS  Andrew Parry MA VetMB CertVDI Diplomate ECVDI  W. Malcolm McKee BVMS MVS MACVSc DSAO 《Veterinary surgery : VS》2013,42(6):751-758

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Comparison of Complication Rates and Clinical Outcome Between Tibial Plateau Leveling Osteotomy and a Modified Cranial Closing Wedge Osteotomy for Treatment of Cranial Cruciate Ligament Disease in Dogs     

Bill Oxley MA VetMB CertSAS  Toby J. Gemmill BVSc MVM DSAS Diplomate ECVS  Alasdair R. Renwick BVMS DSAS  W. Malcolm McKee BVMS MVS MACVSc DSAO 《Veterinary surgery : VS》2013,42(6):739-750

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Serologic prevalence of Dirofilaria immitis,Ehrlichia canis,and Borrelia burgdorferi infections in Brazil     

Labarthe N  de Campos Pereira M  Barbarini O  McKee W  Coimbra CA  Hoskins J 《Veterinary therapeutics : research in applied veterinary medicine》2003,4(1):67-75

Dogs infected with Dirofilaria immitis, Ehrlichia canis, or Borrelia burgdorferi may show nonspecific clinical signs or may be asymptomatic. In Brazil, E. canis and D. immitis infections are frequently diagnosed based on the presence of classical signs; however, serologic tests are seldom performed to confirm the presence of infection. To estimate the seroprevalence of these three canine diseases in Brazil, 2,553 dogs presented at veterinary practices for various tests, routine treatments, or examinations were evaluated by an in-office commercial ELISA test kit (SNAP 3Dx, IDEXX Laboratories). Each dog was examined by the veterinarian, and a whole-blood sample was collected and immediately tested for the simultaneous detection of B. burgdorferi and E. canis antibodies and D. immitis antigen. D. immitis infection was detected in 51 dogs (2.0%) and E. canis antibodies were present in 505 dogs 19.8%). Only one dog tested positive for B. burgdorferi antibodies.  相似文献   

Bilateral fixation of Y-T humeral condyle fractures via medial and lateral approaches in 29 dogs     

McKee WM  Macias C  Innes JF 《The Journal of small animal practice》2005,46(5):217-226

OBJECTIVES: To describe bilateral fixation of Y-T fractures of the humeral condyle via combined medial and lateral approaches, and to determine the technique's clinical and radiographic short-term outcomes. METHODS: Details of 30 consecutive fractures in 29 dogs were reviewed. These included signalment, method of fixation, complications, and follow-up limb function and range of elbow joint motion. RESULTS: The age of the dogs ranged from three months to nine years, and bodyweight ranged from 1.9 to 48 kg. The humeral condyle was reattached to the shaft using medial and lateral bone plates in 18 fractures, a medial plate and lateral Kirschner wire(s) in six fractures, and medial and lateral Kirschner wire(s) in six fractures. Major complications were recorded in four fractures and minor complications in two fractures. Limb function at follow-up was graded as excellent in 12, good in 15 and fair in three fractures. The range of elbow flexion was normal in seven, mildly reduced in 18, moderately reduced in four and severely reduced in one fracture. CLINICAL SIGNIFICANCE: In contrast to the caudal approach, combined medial and lateral approaches decrease the extent of periarticular soft tissue dissection, avoid complications associated with olecranon osteotomy and enable exposure of the entire humeral diaphysis for fixation. Bilateral fixation is likely to be better at counteracting bending and torsional forces compared with unilateral fixation.  相似文献   

Caudal proximal tibial deformity and cranial cruciate ligament rupture in small-breed dogs     

Macias C  McKee WM  May C 《The Journal of small animal practice》2002,43(10):433-438

Eight dogs presented with chronic hindlimb lameness associated with cranial cruciate ligament rupture. Seven were small terriers. A caudal deformity of the proximal tibial shaft, originating at the proximal tibial physis, and an excessive caudal slope of the tibial plateau were present bilaterally in all dogs. The deformity was thought to be responsible for the cranial cruciate ligament failure and poor response to conservative management. Tibial plateau angles were in excess of 26 degrees in all dogs. The lameness was bilateral in three dogs. There was complete cranial cruciate ligament rupture in seven stifles and partial rupture in four. There were no meniscal injuries. Surgical correction resulted in a significant improvement (P<0.0001) in all dogs, with a mean follow-up of 12 months (range three to 24 months). There were no complications.  相似文献   

Thoracolumbar disc disease in large dogs: a study of 99 cases     

Macias C  McKee WM  May C  Innes JF 《The Journal of small animal practice》2002,43(10):439-446

The records of 99 dogs weighing over 20 kg with thoracolumbar disc disease were reviewed. Two types of disc disease were recognised: degenerative nuclear extrusion (n=63) and degenerative annular protrusion (n=36). Sixty-nine per cent of the affected discs were located between T12-T13 and L2-L3. Of the 63 dogs with degenerative nuclear extrusions, 35 were non-ambulatory and seven had no conscious pain perception at the time of presentation. Decompressive surgery was performed in 55 dogs, four dogs were managed non-surgically and three dogs were euthanased. A successful outcome was achieved in 49 (78 per cent) cases as assessed by the authors and in 53 (84 per cent) cases as assessed by the owners. Mean follow-up time was 11.7 months (range 1.5 to 48 months). Five dogs subsequently lost the ability to ambulate on their hindlimbs. Myelographic investigations in three of these dogs revealed a second thoracolumbar degenerative nuclear extrusion. Of the 36 dogs with degenerative annular protrusions, seven were non-ambulatory at the time of presentation. Fifteen cases had multiple protrusions. Twenty dogs were managed non-surgically, 12 surgically and four were euthanased. A successful outcome was achieved in eight (22 per cent) cases as assessed by the authors and in 19 (52 per cent) cases as assessed by the owners. Mean follow-up time was 9.2 months (range 1.5 to 30 months). The outcome of dogs with annular protrusions was significantly worse compared to the outcome of dogs with nuclear extrusions (P<0.001).  相似文献   

Ulnocarpal arthrodesis and limb lengthening for the management of radial agenesis in a dog     

McKee WM  Reynolds J 《The Journal of small animal practice》2007,48(10):591-595

Radial agenesis was diagnosed in a nine-week-old boxer. An ulnocarpal arthrodesis was performed when six months of age to improve limb alignment and enable weight bearing. When the dog was 10 months old, a hinged circular fixator was applied to the limb to gradually lengthen the ulna and correct angular and rotational deformity. The fixator was removed at 16 months of age. At this time, there was 1·7 cm (6 per cent) shortening of the affected antebrachium-metacarpus and 2·6 cm (15 per cent) compensatory overgrowth of the humerus compared with the contralateral limb. Inability to extend the digits was a complication that improved with physiotherapy. Follow-up at 23 months of age showed good limb use when walking and running. Radiographs of the elbow showed periarticular osteophytes and subcoronoid sclerosis of the ulna.  相似文献   

Behavioural interactions of diamondback terrapins with crab pots demonstrate that bycatch reduction devices reduce entrapment          下载免费PDF全文

Rebecca K. McKee  Kristen K. Cecala  Michael E. Dorcas 《水产资源保护:海洋与淡水生态系统》2016,26(6):1081-1089