Recovery and Cryopreservation of Epididymal Sperm of Plains Bison (Bison bison bison) as a Model for Salvaging the Genetics of Wood Bison (Bison bison athabascae)     

LC Aurini  DP Whiteside  BT Elkin  JC Thundathil 《Reproduction in domestic animals》2009,44(5):815-822

The objective of this study was to optimize recovery and cryopreservation of epididymal sperm from plains bison, as a model for wood bison. In Phase 1, cauda epididymides were recovered from bison (n = 14) immediately after slaughter, minced and incubated in Sp-TALPH buffer for 3 h at 36°C. The resulting sperm suspensions were cryopreserved in Triladyl^®, using a protocol for bovine semen. In Phase 2, epididymal sperm were cryopreserved in either Triladyl^® or Andromed^®. The mean (±SD) estimated number of sperm recovered was 468 ± 207 × 10⁶. There was an increase (p < 0.05) in the proportion of sperm with normal morphology between initial recovery and after extension (52.4 ± 4.6 vs 69.7 ± 2.4%), with a concurrent decrease (p < 0.05) in the proportion of sperm with distal droplets. Median values for progressively motile sperm in post-thaw samples (60%) were lower (p < 0.05) than that after extension or after chilling (70% for both). The mean percentages of viable sperm and of sperm with an intact acrosome were lower (p < 0.05) for frozen-thawed samples (38.7 ± 2.8 and 85.2 ± 1.1) compared with extended (66.2 ± 2.2 and 92.4 ± 0.9) or chilled (63.7 ± 2.5 and 90.0 ± 1.0) samples. Rates of cleavage, morulae and blastocyst production were not significantly different for chilled (70.9, 38.7 and 8.0%) vs post-thaw sperm (73.0, 46.0 and 6.3%). There was no significant difference between extenders for most sperm characteristics. In conclusion, we developed a functional protocol for the recovery and cryopreservation of epididymal sperm from plains bison, which may have implications for the genetic preservation of wood bison.  相似文献   

Effect of hCG Treatment on the Oestrous and Ovulation Responses to FSH in Prepubertal Gilts     

R Manjarin  JC Dominguez  MJ Castro  DJ Sprecher  G Cassar  RM Friendship  RN Kirkwood 《Reproduction in domestic animals》2009,44(3):432-434

To ensure sufficient numbers of pregnant females, particularly at hotter times of the year, hormonal induction of gilt oestrus may be necessary. However, the gilt oestrus and ovulation responses to gonadotrophin treatment have often proven unpredictable. The objective of this study was to examine possible reasons for this unpredictability. Prepubertal gilts (approximately 150 days of age, n = 63) were assigned to one of three treatments: injection of 300 IU hCG (n = 15); pre-treatment with 100 mg FSH in polyvinylpyrrolidinone administered as 2 × 50 mg injections 24 h apart, followed by 600 IU eCG at 24 h after the second FSH injection (n = 23); or FSH pre-treatment as above followed by 300 IU hCG at 24 h after the second FSH injection (n = 25). To facilitate oestrus detection, gilts were exposed to a mature boar for 15 min daily for 7 days. Blood samples were obtained on the day of eCG or hCG injection and again 10 days later and gilt ovulation responses determined based on elevated progesterone concentrations. The oestrus responses by 7 days were 6.7%, 17.5% and 64.0% for gilts treated with hCG, FSH + eCG and FSH + hCG, respectively (p < 0.001). The oestrous gilt receiving hCG alone and one oestrous FSH + hCG gilt did not ovulate, all other oestrous gilts ovulated. A further two anoestrous FSH + eCG-treated gilts ovulated. These data suggest that FSH pre-treatment facilitated the development of ovarian follicles to the point where they became responsive to hCG, but had little effect on the response to eCG.  相似文献   

Comparison of transcutaneous ultrasound over the right flank with transrectal ultrasound for pregnancy diagnosis in the dairy cow   总被引：1，自引：1，他引：0  

JC Hunnam  TJ Parkinson  N Lopez-Villalobos  S McDougall 《Australian veterinary journal》2009,87(8):318-322

Objective   Evaluate the sensitivity and specificity of transrectal ultrasound and transcutaneous ultrasonography across the right flank between days 31 and 196 of pregnancy in the dairy cow.
Procedure   Pregnancy status and stage of gestation at date of pregnancy diagnosis by transcutaneous and transrectal ultrasonography were determined for 1570 dairy cattle. Sensitivity and specificity values and probabilities of a correct diagnosis for both techniques were determined and compared. Possible effects of gestational age, herd, method of transcutaneous ultrasound diagnosis, cow age, cow and bull breed, and calf sex on the determination of a correct diagnosis of pregnancy status were evaluated for both techniques.
Results   The overall sensitivity and probability of a correct diagnosis of pregnancy status from days 31 to 196 of gestation were significantly higher when using transrectal ultrasound relative to transcutaneous ultrasound over the right flank. However, the sensitivity of transcutaneous ultrasound changed over the course of pregnancy: it was low (7–8%) before day 84, increased to 89% between days 141 and 154, and was close to 100% thereafter. The specificity of both methods did not differ significantly. Herd and gestational age at date of pregnancy diagnosis were significantly associated with making a correct test result using transcutaneous ultrasound, although only the latter predictor was significantly associated with making a correct test result using transrectal ultrasound.
Conclusion   Transcutaneous ultrasound can not be recommended as an accurate method for early pregnancy diagnosis in the dairy cow, particularly on a whole-herd basis. However, if pregnancy testing is undertaken in mid to late gestation, this technique could provide an accurate and rapid alternative to transrectal ultrasound or manual palpation per rectum.  相似文献   

Association between gestational age and bovine fetal characteristics measured by transcutaneous ultrasound over the right flank of the dairy cow     

JC Hunnam  TJ Parkinson  N Lopez-Villalobos  S McDougall 《Australian veterinary journal》2009,87(9):379-383

Objective   To determine bovine fetal characteristics significantly associated with increasing gestational age as measured via transcutaneous ultrasonography over the right flank.
Methods   The length of gestation at date of pregnancy diagnosis via transcutaneous and transrectal ultrasonography was determined for 224 dairy cattle by estimation from subsequent calving dates. A separate dataset was created for each measurable fetal characteristic (i.e. thoracic diameter, abdominal diameter, umbilical diameter, placentome length and placentome height) and risk factors significantly associated with gestational age at pregnancy diagnosis within each dataset, including the fetal characteristic, were identified.
Results   Abdominal diameter was the most frequently observed fetal characteristic and thoracic diameter was the least. Gestational age at pregnancy diagnosis (d) was significantly associated with fetal thoracic diameter (P < 0.01), abdominal diameter (P < 0.01) and umbilical diameter (P = 0.02) when measured via transcutaneous ultrasound. Within each model, sire breed, dam breed, dam age and/or calf sex were also significantly associated with gestational age. Gestational age at pregnancy diagnosis was not significantly associated with either placentome height or length (P > 0.05).
Conclusion   Fetal thoracic diameter, abdominal diameter and umbilical diameter were found to be significantly associated with gestational age between approximate days 73 to 190 of gestation. Transcutaneous ultrasonography may prove a useful method of estimating gestational age in the absence of accurate breeding records.  相似文献   

Infertility associated with the absence of endometrial progesterone receptors in a bitch          下载免费PDF全文

JC Dockweiler  B Cossic  CG Donnelly  RO Gilbert  E Buckles  SH Cheong 《Reproduction in domestic animals》2017,52(1):174-178

A three‐year‐old intact female Old English sheepdog was presented for evaluation of infertility. A uterine biopsy was performed during dioestrus, and the microscopic appearance was inconsistent with progesterone stimulation; the glands were sparse, simple and failed to show coiling, while the glandular epithelium was cuboidal instead of columnar. There was very little evidence of glandular activity. Due to the inappropriate appearance of the glands for the stage of the cycle, immunohistochemistry for progesterone receptors was performed. No progesterone receptor‐positive immunoreactivity was identified in the endometrial luminal epithelium, glandular epithelium or stroma. Weak intranuclear immunoreactivity was identified within the smooth muscle cells of the myometrium. The absence of progesterone receptors within the endometrial glands is the most likely explanation for the abnormal appearance of the endometrium and for this bitch's infertility. To our knowledge, this is the first report of endometrial progesterone receptor absence in a bitch.  相似文献   

The "expanding universe" of piroplasms     

Criado-Fornelio A  Gónzalez-del-Río MA  Buling-Saraña A  Barba-Carretero JC 《Veterinary parasitology》2004,119(4):337-345

The present paper is the continuation of our previous studies dealing with the genetic characterization of piroplasmid species found in southern Europe. We report in this work new data concerning sequences of the 18s rRNA gene in Spanish piroplasms not studied (or not totally sequenced) in our former surveys. Molecular data analysis indicated that Spanish Cytauxzoon felis (cat isolate) has 98% identity with Cytauxzoon sp. from Mongolia and 95% identity compared to African C. felis. There are at least two main genetic variants of Babesia caballi in Spain: The first variety (isolate Spain 1) shows a relatively low homology with the African genotype (97% identity). The second variety (represented by two isolates, Spain 2 and Spain 3, differing by a single base) shows high genetic similarity with the African genotype (99.7-100% identity). There are also two genetic variants of Babesia equi (isolates Spain 1 and Spain 2, differing by four bases) in Spain, sharing 99% identity with the African genotype. At least one of them (Spain 1) can infect dogs. All of the phylogenetic analysis procedures employed indicated that Spanish isolates of C. felis, B. caballi (Spain 1) and B. equi (Spain 1 and Spain 2) are genetically different from their African relatives, all those dichotomies showing very high bootstrap support. Nonetheless, the lack of information on their morphology and the fact that the sequences were obtained in a single isolate preclude any conclusion about their definitive taxonomic status.  相似文献   

Presence of Mycoplasma haemofelis,Mycoplasma haemominutum and piroplasmids in cats from southern Europe: a molecular study     

Criado-Fornelio A  Martinez-Marcos A  Buling-Saraña A  Barba-Carretero JC 《Veterinary microbiology》2003,93(4):307-317

Clinical symptoms produced by Mycoplasma spp. and piroplasmids in cats are sometimes similar. Diagnosis of these pathogens is difficult by microscopic procedures and molecular methods have been used as an alternative. We present in this work, the development of new molecular procedures for diagnosis of the aforementioned organisms, together with a molecular characterization of isolates found in southern European cats.A single PCR-RFLP procedure was designed for diagnosis of Mycoplasma spp. and a seminested PCR-RFLP was designed for diagnosis of piroplasmids. The 16S or 18S rRNA genes of isolates found in clinical samples were partially sequenced in all positive cases.Mycoplasma spp. was detected in 9 (30%) out of 30 symptomatic cats from Spain. Sequencing indicated that 66.6% of these isolates can be ascribed to Mycoplasma haemofelis and only 33.3% to Mycoplasma haemominutum. Partial 16S rRNA sequences obtained in Spanish isolates were very similar to those previously published from the UK and the USA.The presence of piroplasmids (Babesia and Theileria spp.) was studied in 16 cats from Spain (n=13) and Portugal (n=3). Animals analyzed were 10 cats with immunosuppressive viral infection (either FeLV or FIV), 5 asymptomatic cats and 1 cat with Babesia-compatible symptoms. Asymptomatic cats were all PCR-negative. Partial sequencing of 18S rRNA gene demonstrated that the Babesia-symptomatic cat was infected with Babesia canis canis whereas 3 (30%) out of the 10 cats with immunosuppressive viral infection were coinfected with piroplasmids (1 with B. canis canis, 1 with Theileria annae, and 1 with B. canis canis and T. annae both).  相似文献   

Molecular characterization of a Babesia gibsoni isolate from a Spanish dog   总被引：1，自引：0，他引：1  

Criado-Fornelio A  Gónzalez-del-Río MA  Buling-Saraña A  Barba-Carretero JC 《Veterinary parasitology》2003,117(1-2):123-129

Babesia gibsoni is a morphologically small Babesia species that infects dogs. Molecular techniques have shown that some small Babesia sp. recently described in canids are not related to the original B. gibsoni and they should be assigned to separate taxons. Although the 18s rRNA gene of true B. gibsoni isolates has been studied in the USA, Asia and Australia, no molecular data on the presence and genetic characteristics of B. gibsoni in Europe are available. Blood collected from a Babesia-symptomatic dog from Spain was used for DNA diagnosis by seminested PCR. DNA amplification was positive and the complete 18s rRNA gene of the dog isolate was sequenced, showing 98% homology with B. gibsoni (isolate Asia 1). Evidence from phylogenetic analysis indicated that: The Spanish isolate unambiguously belongs to the B. gibsoni group. The B. gibsoni complex might be diphyletic. In the absence of genetic data from African isolates of B. gibsoni, Asia seems to be the most likely geographical location of origin.  相似文献   

Molecular studies on Babesia,Theileria and Hepatozoon in southern Europe. Part I. Epizootiological aspects     

Criado-Fornelio A  Martinez-Marcos A  Buling-Saraña A  Barba-Carretero JC 《Veterinary parasitology》2003,113(3-4):189-201

Molecular epizootiology of piroplasmids (Babesia spp., Theileria spp.) and Hepatozoon canis was studied in mammals from southern Europe (mainly from Spain, but also from Portugal and France). Partial amplification and sequencing of the 18s rRNA gene was used for molecular diagnosis. In some particular cases (B. ovis and B. bovis) the complete 18s rRNA gene was sequenced. Blood samples were taken from domestic animals showing clinical symptoms: 10 dogs, 10 horses, 10 cows, 9 sheep and 1 goat. In addition, DNA samples were isolated from blood of 12 healthy dogs and from spleen of 10 wild red foxes (Vulpes vulpes). The results of the survey were the following: Piroplasmid infections: Approximately from 50 to 70% of wild or domestic mammals (symptomatic) were infected.Piroplasmids detected in ruminants were:COW: B. bovis, T. annulata and Theileria sp. (type C). Sheep and goat: B. ovis. Piroplasmids present in canids were: Babesia canis vogeli, Babesia canis canis, Theileria annae and B. equi. The only piroplasmid found in asymptomatic dogs was B. equi. Piroplasmids found in horse were: B. equi and B. canis canis.H. canis infections in canids: H. canis was absent of domestic dog samples, whereas all foxes studied were infected by this protozoa.Genetic analysis showed that most of piroplasmid and Hepatozoon isolates from southern Europe matched unambigously with previously described species, as demonstrated by the high level sequence identity between them, usually between 99 and 100%. Minor differences, usually detected in hypervariable regions of 18s rRNA gene are probably due to strain variations or rare genetic polymorphisms. A possible exception was B. bovis, which shows a relatively lower degree of homology (94%) with regard to other B. bovis isolates from several countries. The same is true for B. ovis, that showed a 94% identity with regard to Babesia sp. from South African cow and a 92% with rapport to B. bovis from Portugal.  相似文献   

The use of vascular access ports in nonanesthetized green iguanas (iguana iguana) to collect baseline arterial blood gas parameters.     

JC Hess  KA Grimm  GJ Benson  WA Tranquilli  R Sarr 《Veterinary anaesthesia and analgesia》2005,32(4):18-18

The green iguana, Iguana iguana, is used as a model in reptile anesthesia research because of its size, availability, and the body of knowledge characterizing its physiology. Arterial blood gas values in nonanesthetized green iguanas have not been determined because of the technical difficulty involved. Vascular access port (VAP) placement to facilitate blood sampling has been described in other species, but not lacertilians. This abstract describes the technique for placement of VAPs and the values for arterial blood gas parameters in seven 1 kg adult green iguanas. Using sterile technique, a 1.5 cm incision was made on the lateral side of the neck. Blunt dissection ventral to the external jugular vein revealed the internal and external carotid arteries near their bifurcation. The catheter was inserted into the internal carotid artery and then guided to the common carotid artery. The other end of the catheter was tunneled below the skin to a subcutaneous location, caudal‐dorsal to the iSPSilateral scapula. The skin was closed and the port was flushed twice a week with heparinized saline. Post‐operatively, the VAPs were well tolerated by the iguanas. Difficulties included port disconnection (n = 1), inability to aspirate blood after a few weeks (n = 2), and infection (n = 1). The iguanas were breathing room air prior to and during blood collection. From the five functional VAPs, the blood pH, PCO₂, PO₂, HCO^‐₃, and BE (measured at 37 °C) were 7.45 ± 0.06; 37.5 ± 7.0 mm Hg, 99.0 ±16.6 mm Hg, 25.4 ± 2.5 mmol L^–1, and 1.5 ±2.4 mmol L^–1 respectively (mean ± SD). VAPs can be successfully used to facilitate collection of arterial blood gas samples in green iguanas. These values are similar to those reported for most mammalian species. This technique should facilitate research in anesthesiology and respiratory physiology of iguanas and other lacertilians.  相似文献