Oxygen Concentration and Cysteamine Supplementation During In vitro Production of Buffalo (Bubalus bubalis) Embryos Affect mRNA Expression of BCL‐2, BCL‐XL,MCL‐1, BAX and BID     

G Elamaran  KP Singh  MK Singh  SK Singla  MS Chauhan  RS Manik  P Palta 《Reproduction in domestic animals》2012,47(6):1027-1036

This study examined the effects of O₂ concentration (5% vs 20%) during in vitro maturation (IVM), fertilization (IVF) and culture (IVC) or supplementation of IVM and IVC media with cysteamine (50 and 100 μm , respectively; IVM, IVF and IVC carried out in 20% O₂), on blastocyst rate and relative mRNA abundance of some apoptosis‐related genes measured by real‐time qPCR in immature and in vitro‐matured buffalo oocytes and in embryos at 2‐, 4‐, 8‐ to 16‐cell, morula and blastocyst stages. The blastocyst rate was significantly higher (p < 0.05) while the percentage of TUNEL‐positive cells was significantly lower (p < 0.05) under 5% O₂ than that under 20% O₂. The mRNA expression of anti‐apoptotic genes BCL‐2 and MCL‐1 was significantly higher (p < 0.05) and that of pro‐apoptotic genes BAX and BID was lower (p < 0.05) under 5% O₂ than that under 20% O₂ concentration at many embryonic stages. Following cysteamine supplementation, the blastocyst rate and the relative mRNA abundance of BCL‐XL and MCL‐1 was significantly higher (p < 0.05) and that of BAX but not BID was lower (p < 0.05) at many stages of embryonic development, although it did not affect the percentage of TUNEL positive cells in the blastocysts significantly. The mRNA expression pattern of these genes during embryonic development was different in 5% vs 20% O₂ groups and in cysteamine supplemented vs controls. At the 8‐ to 16‐cell stage, where developmental block occurs in buffalo, the relative mRNA abundance of BCL‐2 and MCL‐1 was highest under 5% O₂ concentration and that of BAX and BID was highest (p < 0.05) under 20% O₂ concentration. These results suggest that one of the mechanisms through which beneficial effects of low O₂ concentration and cysteamine supplementation are mediated during in vitro embryo production is through an increase in the expression of anti‐apoptotic and a decrease in the expression of pro‐apoptotic genes.  相似文献   

Conception rates to fixed-time artificial insemination of two oestrus synchronisation programmes in dairy heifers     

SK Sahu  TJ Parkinson  RA Laven 《New Zealand veterinary journal》2013,61(3):158-161

AIM: To evaluate the conception rate to fixed-time artificial insemination (FTAI) of two oestrus synchronisation programmes in dairy heifers on eight farms over 2 years.

METHODS: The study was conducted in 2008 and 2010 on eight farms near Palmerston North, New Zealand. Nulliparous Friesian and Friesian×Jersey heifers (13–15 months of age) were randomly allocated to one of two oestrus synchronisation programmes. Group 1 (GPG+P4; n=330), received gonadotrophin-releasing hormone (GnRH) I/M on Day 0, a progesterone (P4)-releasing intravaginal device from Days 0–7, prostaglandin F_2α (PGF) I/M on Day 7 and a second dose of GnRH at the time of FTAI on Day 9. The second group (P4+PGF; n=343) received a P4-releasing intravaginal device from Days 0–7, PGF on Day 6 and FTAI on Day 9. Pregnancy was diagnosed from Days 42–52 by transrectal ultrasonography.

RESULTS: The overall conception rate was 52.4% and 54.8% for the GPG+P4 and P4+PGF groups, respectively. The odds of conception for the two treatments were not different (OR=0.90; 95% CI=0.67–1.23), nor was there any difference between groups in different years (p=0.58). Farm affected conception rate (p=0.002), but there was no interaction with treatment (p=0.92) .

CONCLUSIONS: This study has shown that an alternative synchronisation programme can produce similar results in terms of conception rate to the GPG+P4 treatment, currently commonly used in heifers. More research is required to establish whether other modifications to the GPG+P4 programme can produce similar results at lower costs, and to identify and quantify farm factors which affect the economic benefit of heifer synchronisation.

CLINICAL RELEVANCE: This study indicated that synchronising heifers with P4 and PGF resulted in conception rates equivalent to those resulting from a GPG+P4 treatment, but with reduced drug costs. However, because heifers in the GPG+P4 group received the second GnRH injection at the time of AI, they needed only three yardings as opposed to the four required for the heifers treated with P4 and PGF. Thus, the choice of programme for an individual farm will depend on that farm's circumstances, in particular the cost of yarding the heifers.  相似文献   

Effect of Supplementation of Taurine or Trehalose in Extender on Immunolocalization of Tyrosine Phosphoproteins in Buffalo and Cattle (Karan Fries) Cryopreserved Spermatozoa     

R Kumar  VK Singh  S Chhillar  SK Atreja 《Reproduction in domestic animals》2013,48(3):407-415

The present study assessed the effects of incorporation of Taurine or Trehalose in extender on immunolocalization of tyrosine phosphoproteins, Cryocapacitation and other sperm quality parameters (motility, viability and membrane integrity) in post‐thawed sperm from Buffalo (Murrah) and Cattle (Karan Fries). Six ejaculates from six individual bulls from both species were chosen at random and split into four aliquots: one aliquot without dilution (fresh sample), another diluted in egg yolk tris‐citrate (EYTC) extender and the rest of aliquots with EYTC dilution supplemented with taurine (50 mm ) or trehalose (100 mm ), respectively, and cryopreserved. Following cryopreservation, semen were thawed and assessed for standard semen quality parameters. Extent of capacitation in cryopreserved spermatozoa was measured by inducing in vitro acrosome reaction followed by dual staining. Immunolocalization of tyrosine phosphoproteins was carried out by immunocytochemistry using primary antibody clone pT‐154 (anti‐phosphotyrosine antibody) and FITC‐conjugated secondary antibody. Immunofluorescent signals were analysed for level of protein tyrosine phosphorylation in spermatozoa. Post‐thaw semen evaluation showed supplementation of taurine or trehalose to EYTC extender significantly (p < 0.05) increased motility, viability and membrane integrity of spermatozoa in both species. Percentage of cryocapacitated spermatozoa was significantly (p < 0.05) higher in cattle as compared to buffalo and degree of cryocapacitaion of spermatozoa decreased significantly (p < 0.05) upon supplementation of additives in both the species. It was also found that tyrosine phosphoproteins were localized differentially in fresh and cryopreserved spermatozoa. Supplementation of taurine or trehalose to freezing extender changed the localization of tyrosine phosphoproteins in cryopreserved spermatozoa similar to fresh in both the species. The results obtained clearly indicated that supplementation of taurine or trehalose to EYTC prior to cryopreservation improves Buffalo and Cattle sperm quality in terms of cryocapacitation and immunolocalization of tyrosine phosphoproteins during freezing–thawing process.  相似文献   

Molecular expression of caprine estrogen receptor gene 1 in reproductive and non‐reproductive tissues          下载免费PDF全文

S Saraswat  PK Rout  SD Kharche  SK Jindal  AK Goel 《Reproduction in domestic animals》2016,51(6):1049-1054

During the last decades, physiological effects of oestrogens have been increasingly explored by scientists and biotechnologists. Estrogens exert a wide range of effects on a large variety of cell types. Oestrogen and its receptors are essential for sexual development and reproduction. Estrogen receptor alpha is a nuclear receptor activated by the hormone oestrogen. In male, ERα is encoded by the gene estrogen receptor gene 1 (ESR1), responsible for better fertility. The ESR1 is involved in the reabsorption of luminal fluid during the transit of spermatozoa from the testis to the head of the epididymis which is important for their survival and maturation during epididymal storage. The absence of ESR1 leads to reduced epididymal sperm content, reduced sperm motility and fertilizing ability. Therefore, this is a good startby to study the expression pattern of estrogen receptor 1 gene in high‐fertile (G1) and low‐fertile (G2) bucks of Jamunapari and Barbari breeds identified on the basis of seminal quality traits and fertility trials. RNA was extracted from the tissues by TRIzol method. The identification and expression pattern of caprine ESR1 gene was analysed by real‐time PCR (Roche LC‐480). Our work shows that the relative quantification by RT‐PCR indicates more fold in head of epididymis as compared to spleen of caprine ESR1 gene. Furthermore, the RT‐PCR indicated that fertile bucks of Jamunapari breed have more fold value as compared to Barbari breed in respect of reproductive organ.  相似文献   

Effects of low‐density lipoproteins as additive on quality parameters and oxidative stress following cryopreservation of mithun (Bos frontalis) spermatozoa          下载免费PDF全文

P Perumal  SK Srivastava  SK Ghosh  KK Baruah  S Bag  JS Rajoria  K Kumar  C Rajkhowa  M Pande  N Srivastava 《Reproduction in domestic animals》2016,51(5):708-716

Artificial breeding of mithun poses several challenges including lack of standard protocol for cryopreservation of spermatozoa. This is further complicated by harmful effects of hen's egg yolk (EY) as additive in extender. Purified low‐density lipoproteins (LDL) extracted from EY have been shown as beneficial over EY extender for long‐term semen storage in several species. This investigation explored use of LDL versus EY on semen quality and oxidative stress following freezing–thawing of spermatozoa. A total of 25 of 50 ejaculates based on biophysical parameters were selected for the experiment. After diluting with the Tris‐citrate‐glycerol (TCG) extender, each sample was split into three equal aliquots: Group I, control, EY; Group II and Group III contained 8% and 10% purified LDL, respectively. Frozen–thawed samples were evaluated for motility parameters (progressive, and in the bovine cervical mucus penetration test [BCMPT]), viability, sperm and nuclear abnormality, acrosome integrity, and enzymatic (leakage of intracellular contents) and biochemical (oxidative stress) profiles and in vitro fertility (IVF) assay. Study revealed a significant (p < .05) improvement in viability, sperm and nuclear abnormality, acrosome integrity, motility (progressive and in cervical mucus), cholesterol content, and reduction in the leakage of intracellular enzymes in Group II. Moreover, intactness of acrosome and biochemical membranes was protected significantly (p < .05) in addition to significant (p < .05) improvement in binding per cent and binding index in IVF assay in extender containing 8% LDL. These results demonstrate that although cryopreservation of mithun's spermatozoa in EY was comparable with other species, addition of 8% LDL holds a clear advantage over EY or 10% LDL.  相似文献   

Serum MX2 Protein as Candidate Biomarker for Early Pregnancy Diagnosis in Buffalo          下载免费PDF全文

L Buragohain  R Kumar  T Nanda  SK Phulia  AK Mohanty  S Kumar  S Balhara  SPS Ghuman  I Singh  AK Balhara 《Reproduction in domestic animals》2016,51(4):453-460

  相似文献   

Conventional and future diagnostics for avian influenza     

Charlton B  Crossley B  Hietala S 《Comparative immunology, microbiology and infectious diseases》2009,32(4):341-350

The significant and continued transboundary spread of Asian avian influenza H5N1 since 2003, paired with documented transmission from avian species to humans and other mammals, has focused global attention on avian influenza virus detection and diagnostic strategies. While the historic and conventional laboratory methods used for isolation and identification of the virus and for detection of specific antibodies continued to be widely applied, new and emerging technologies are rapidly being adapted to support avian influenza virus surveillance and diagnosis worldwide. Molecular tools in particular are advancing toward lab-on-chip and fully integrated technologies that are capable of same day detection, pathotyping, and phylogenetic characterization of influenza A viruses obtained from clinical specimens. The future of avian influenza diagnostics, rather than moving toward a single approach, is wisely adopting a strategy that takes advantage of the range of conventional and advancing technologies to be used in "fit-for-purpose" testing.  相似文献   

A review of thermal stress in cattle     

RW Shephard  SK Maloney 《Australian veterinary journal》2023,101(11):417-429

Cattle control body temperature in a narrow range over varying climatic conditions. Endogenous body heat is generated by metabolism, digestion and activity. Radiation is the primary external source of heat transfer into the body of cattle. Cattle homeothermy uses behavioural and physiological controls to manage radiation, convection, conduction, and evaporative exchange of heat between the body and the environment, noting that evaporative mechanisms almost exclusively transfer body heat to the environment. Cattle control radiation by shade seeking (hot) and shelter (cold) and by huddling or standing further apart, noting there are intrinsic breed and age differences in radiative transfer potential. The temperature gradient between the skin and the external environment and wind speed (convection) determines heat transfer by these means. Cattle control these mechanisms by managing blood flow to the periphery (physiology), by shelter-seeking and standing/lying activity in the short term (behaviourally) and by modifying their coats and adjusting their metabolic rates in the longer term (acclimatisation). Evaporative heat loss in cattle is primarily from sweating, with some respiratory contribution, and is the primary mechanism for dissipating excess heat when environmental temperatures exceed skin temperature (~36°C). Cattle tend to be better adapted to cooler rather than hotter external conditions, with Bos indicus breeds more adapted to hotter conditions than Bos taurus. Management can minimise the risk of thermal stress by ensuring appropriate breeds of suitably acclimatised cattle, at appropriate stocking densities, fed appropriate diets (and water), and with access to suitable shelter and ventilation are better suited to their expected farm environment.  相似文献   

Pharmacokinetics and estimated bioavailability of amoxicillin in mares after intravenous, intramuscular, and oral administration     

W D Wilson  M S Spensley  J D Baggot  S K Hietala 《American journal of veterinary research》1988,49(10):1688-1694

The pharmacokinetics and estimated bioavailability of amoxicillin were determined after IV, intragastric, and IM administration to healthy mares. After IV administration of sodium amoxicillin (10 mg/kg of body weight), the disposition of the drug was best described by a 2-compartment open model. A rapid distribution phase was followed by a rapid elimination phase, with a mean +/- SD half-life of 39.4 +/- 3.57 minutes. The mean volume of distribution was 325 +/- 68.2 ml/kg, and the mean body clearance was 5.68 +/- 0.80 ml/min.kg. It was concluded that frequent IV administration of sodium amoxicillin would be required to maintain therapeutic plasma concentrations of amoxicillin, and thus, the use of this dosage form should be limited to the initiation of treatment or to intensive care situations. After intragastric administration of amoxicillin trihydrate (20 mg/kg), 5% cherry-flavored suspension, the drug was rapidly, but incompletely, absorbed and rapidly eliminated (mean half-life of the decline phase of the plasma amoxicillin concentration-time curve, 51 minutes). The mean estimated bioavailability (fractional absorption) of the administered dose was 10.4%, and the mean peak plasma amoxicillin concentration was 2.73 micrograms/ml at 1.5 hours after dosing. In one horse with clinical signs of abdominal discomfort and diarrhea, the absorption of amoxicillin from the gastrointestinal tract was delayed and the fraction absorbed was increased. It was concluded that this oral dosage form could be recommended only for the treatment of infections caused by bacteria that are highly susceptible to amoxicillin, that frequent dosing would be necessary, and that absorption may be inconsistent in horses with gastrointestinal disease.(ABSTRACT TRUNCATED AT 250 WORDS)  相似文献   

Prevaccination bovine viral diarrhea virus titers and subsequent reproductive performance in dairy heifers.          下载免费PDF全文

P Michel  M Thurmond    S Hietala 《Canadian journal of veterinary research》1993,57(4):236-240

The study was conducted to determine if there was a relationship between prevaccination bovine viral diarrhea virus (BVDV) titers in 14 month-old dairy heifers and time open during first lactation, length of the conception interval, fetal survival during the second pregnancy, and age at culling. Possible associations were analyzed using nonparametric survival methods and survival distributions were described using the product-limit (Kaplan Meier) methods. Regression analysis was used to estimate the correlation between pre- and postvaccination BVDV titers three years after vaccination. Evidence of exposure to BVDV prior to 14 months of age was demonstrated by serum-virus-neutralization titers greater than 1:4 for 97% and greater than or equal to 1:128 for 67% of the 72 heifers followed. No correlation was found for 38 pairs of prevaccination and postvaccination titers (p = 0.76). The risk of being culled was high for heifers (n = 41) with very low (1:2-1:8) or very high (> or = 1:256) titers, but low for heifers (n = 31) with moderate (1:16-1:128) prevaccination titers (p = 0.098). Risk of subsequent fetal loss was high for heifers (n = 30) with very low (1:2-1:16) or very high (> or = 1:256) prevaccination titers, compared to heifers (n = 24) with moderate (1:32-1:128) titers (p = 0.084). These findings suggest that prevaccination exposure to BVDV eliciting either a very weak or a very strong serological response may contribute to subsequent reproductive inefficiency and an increased risk of culling.  相似文献