磷酸泰乐菌素颗粒剂生物效价测定方法的建立及验证

建立了磷酸泰乐菌素颗粒剂的生物效价测定方法，并对新建立的测定方法进行了验证，验证内容包括：方法对照试验、精密度试验、线性及线性范围试验、回收率试验。验证结果证明新方法能准确测定磷酸泰乐菌素颗粒剂的效价单位。     

Influence of Staining Method on the Values of Avian Sperm Head Morphometric Variables

Computer‐assisted systems for the assessment of sperm morphometry (ASMA systems) have been used successfully with several mammalian species. Unfortunately, they have so far been of little use for assessing bird semen, a consequence of the filiform shape of avian spermatozoa. This study compares two staining techniques (Hemacolor^® and aniline blue staining) for the morphometric analysis of rooster and red‐legged partridge spermatozoa as part of a computer‐assisted light microscopy method. For both species, Hemacolor^® staining provided a significantly higher percentage of measurable cells (93.7 ± 11.7% in roosters and 71.9 ± 15.3% in red‐legged partridges). Hemacolor^® also showed greater repeatability (lower coefficients of variation) for length and area in roosters' sperm and for width in the case of red‐legged partridge's sperm. In the roosters, the Hemacolor^® technique returned significantly (p < 0.05) larger sperm head width and area values than did the aniline blue technique, while the latter resulted in greater sperm head length values (p < 0.05). In the red‐legged partridge, no differences were seen in the results for sperm head width and area provided by the two techniques, but aniline blue staining was associated with longer length measurements. In conclusion, the morphometric values recorded differed depending on the staining method and species. However, the Hemacolor^® technique might be deemed the more appropriate for computerized sperm assessment systems as it provides larger percentages of measureable cells and shows greater repeatability.     

Effect of the Interaction Between Cryoprotectant Concentration and Cryopreservation Method on Frozen/Thawed Chicken Sperm Variables

This work examines the effect of the interaction between different concentrations of two cryoprotectants – glycerol (GLY) and dimethylacetamide (DMA) – and two methods of cryopreservation – pellets produced by plunging into liquid nitrogen and gradual in‐straw freezing – on frozen/thawed chicken sperm variables. Sperm was cryopreserved using: (i) 6% DMA, following the in‐straw and the pellet methods (ii) 11% GLY, following the in‐straw and the pellet methods; and (iii) 8% GLY in the in‐straw method and 3% DMA in the pellet method (i.e. reduced cryoprotectant concentrations). When 6% DMA was used as the cryoprotectant, no differences were seen between the in‐straw and pellet methods in terms of frozen/thawed sperm variables or fertility (10.8% and 12.8%, respectively). The viability and motility variables of the frozen/thawed sperm produced using the in‐straw method with 11% GLY were higher (p < 0.05) than those recorded for the sperm preserved using the same cryoprotectant and concentration in the pellet method. However, fertility was extremely low in both groups (2.1% and 4.2% for the in‐straw and pellet methods, respectively). Finally, the use of 8% GLY in the in‐straw method returned higher sperm viability, intact acrosome and motility values than the use of 3% DMA in the pellet method (p < 0.01). No differences were seen, however, in the fertility results obtained (28.8% and 25.0%, respectively). These results suggest that cryoprotectant concentrations can be reduced and still provide acceptable fertility rates.     

A Pilot Study to Compare Oxidative Status between Organically and Conventionally Managed Dairy Cattle During the Transition Period

The aim of this study was to assess the redox balance of organically managed dairy cattle (OMC; n = 40) during the transition period and to compare this with conventionally managed cattle (CMC; n = 22). Serum samples of dairy cows from two organic and one conventional farm were taken. Markers of oxidants production [reactive oxygen species] and total serum antioxidant capacity were measured in four different production stages: (i) far‐off dry (2 to 1 months before calving; 44 samples in CMC and 48 in OMC); (ii) close‐up dry (1 month until 3 days before calving; 44 CMC; 54 OMC); (iii) fresh (3 days to +1 month after calving; 44 CMC; 49 OMC); and (iv) peak of lactation (+1 to +3 months; 71 CMC; 78 OMC). Values were compared between production stages and against a metabolic baseline status (4th–5th month of pregnancy; 40 CMC; 30 OMC). Our results indicated that throughout the periparturient period, OMC had lower concentrations of reactive oxygen species, but also a lower antioxidant capacity than CMC. Indeed, when the two components of the redox balance were assessed together through the Oxidative Stress index, the values of this parameter were higher for OMC than for CMC, thereby implying a higher risk of oxidative stress. Therefore, further larger studies are needed to confirm the current observations, as organically reared animals might be exposed to a lack of antioxidants supply.     

Equine castration: review of anatomy, approaches, techniques and complications in normal, cryptorchid and monorchid horses

Complications associated with equine castration are the most common cause of malpractice claims against equine practitioners in North America. An understanding of the embryological development and surgical anatomy is essential to differentiate abnormal from normal structures and to minimise complications. Castration of the normal horse can be performed using sedation and regional anaesthesia while the horse is standing, or under general anaesthesia when it is recumbent. Castration of cryptorchid horses is best performed under general anaesthesia at a surgical facility. Techniques for castration include open, closed and half-closed techniques. Failure of left and right testicles to descend occurs with nearly equal frequency, however, the left testicle is found in the abdomen in 75% of cryptorchid horses compared to 42% of right testicles. Bilateral cryptorchid and monorchid horses are uncommon. Surgical approaches described for the castration of cryptorchid horses include an inguinal approach with or without retrieval of the scrotal ligament, a parainguinal approach, or less commonly a suprapubic paramedian or flank approach. Laparoscopic castration of cryptorchid horses has recently been described but the technique has limited application in practice at this time. A definitive diagnosis of monorchidism can only be made after surgical exploration of the abdomen, removal of the normal testis and hormonal testing. Hormonal assays reported to be useful include analysis of basal plasma or serum testosterone or oestrone sulphate concentrations, testosterone concentrations following hCG stimulation, and faecal oestrone sulphate concentrations. Reported complications of castration include postoperative swelling, excessive haemorrhage, eventration, funiculitis, peritonitis, hydrocele, penile damage and continued stallion-like behaviour.     

Concentrate supplementation, and the herbage intake and milk production of heifers grazing Cenchrus ciliaris

Friesian heifers grazing Cenchrus ciliaris cv. Biloela were supplemented with 0, 3 or 6 kg concentrates daily during weeks 10–34 (±1·7) of lactation during either the rainy or the dry season. The overall responses to concentrate were identical between seasons at 0·27 kg extra milk and solids corrected milk per kg. Supplementation increased total feed intake and modified the grazing behaviour of cows. For each kg concentrate organic matter eaten, herbage organic matter intake was reduced by 0·64 and 0·42 kg in the rainy and dry seasons respectively and the time spent grazing by 11 min. Higher intakes in the dry season were the result of an increased rate of biting and were reflected in liveweight change but not milk yield.     

Clearance of para-aminohippuric acid in wethers consuming locoweed

AIM: To validate the use of para-aminohippuric acid (PAH) as a marker for measuring blood flow in wethers consuming a mixed diet of locoweed and blue grama hay.

METHODS: Fourteen sheep, stratified by bodyweight (BW), were assigned to one of three treatments: 0.8 mg swainsonine (SW)/kg BW (HI), 0.2 mg SW/kg BW (LO), and no SW (Control). Sheep were fed various ratios of locoweed and blue grama hay to deliver SW treatments, for 28 days prior to infusion of PAH. Concentrations of SW and activities of alkaline phosphatase (Alk-P) and aspartate aminotransferase (AST) in serum were measured to confirm exposure to SW and subclinical intoxication. A single 20-ml injection of 5% PAH was delivered into the jugular vein after subclinical intoxication had been achieved. Blood samples were collected and serum analysed for PAH immediately prior to injection, then every 5 min from 5–30 min, and every 10 min from 30–60 min, following injection of PAH.

RESULTS: Effective delivery of SW was evident from the greater concentrations of SW measured in the serum of HI compared with LO animals (p<0.05). No significant differences were detected in the rate of elimination (range 0.097–0.108 L/min), elimination half-life (range 6.62–7.24 min), apparent volume of distribution for the central compartment (range 7.14–9.72 L), and clearance (range 0.73–0.92 L/min) of PAH, between treatments.

CONCLUSIONS: Subclinical intoxication with SW did not affect the pharmacokinetics of PAH. Thus, use of downstream dilution of PAH is a valid method to determine the rate of blood flow in nutrient flux experiments that involve consumption of locoweed.     

The effects of change in herbage mass on rates of herbage growth and senescence in mixed swards

A study was conducted to determine the influence of a change in herbage mass on rates of herbage growth (G), senescence (S) and net production of green herbage (NP) in swards of Lolium perenne L., Poa annua L. and Trifolium repens L. under continuous stocking with sheep. Swards were maintained at either high (1700–1900 kg OM ha⁻¹; H) or low (700–900 kg OM ha⁻¹; L) herbage mass from April to 20 August. From 20 August to 13 September half of the L sward was permitted to grow until a herbage mass similar to that of the H sward was achieved (LH), and at the same time half of the H sward was grazed down until a herbage mass similar to that of the L sward was achieved (HL). The other half of each sward was held at constant mass (LL and HH). Estimates of G, S and NP were obtained for each plant species over a 2-week period from 17 September to 1 October and estimates of species population densities were also made.
The combined species rates of G and S per unit area were highest in treatment LH and lowest in treatment HL, whilst the rates for the LL and HH swards were intermediate. The rate of NP was similar in the LL, HH and LH treatments (34.5, 29.3, 33.6 ± 6.2 kg DM ha⁻¹d⁻¹) but was lower in the HL sward (100 ± 6.2). The effects of alteration of herbage mass on individual tiller or stolon G and S rates were rapid but population density changes were slower.
Within the limited conditions of this trial it was not possible to increase NP by manipulation of herbage mass under continuous stocking management but the results indicate that NP can be reduced in the short term if a sward of high herbage mass and low population density is grazed hard.     

Improvement of 2D‐PAGE Resolution of Human,Porcine and Equine Follicular Fluid by Means of Hexapeptide Ligand Library

The major challenge of follicular fluid proteomic analysis is the presence of high‐abundance proteins that originate from plasma. These proteins can prevent the detection of lower abundant ones, produced locally by follicle cells and that may have important roles in follicular activity. In this study, the novel technology called hexapeptide ligand library was evaluated to enrich the low‐abundance proteins in follicular fluid of human (HFF), porcine (PFF) and equine (EFF) prior 2D‐PAGE. Our results showed that the new strategy enabled detection of many new protein spots, increased resolution and highly improved the intensity of low‐abundance proteins by 2D‐PAGE.     

Calcium, calmodulin, and CaMKII requirement for initiation of centrosome duplication in Xenopus egg extracts

Aberrant centrosome duplication is observed in many tumor cells and may contribute to genomic instability through the formation of multipolar mitotic spindles. Cyclin-dependent kinase 2 (Cdk2) is required for multiple rounds of centrosome duplication in Xenopus egg extracts but not for the initial round of replication. Egg extracts undergo periodic oscillations in the level of free calcium. We show here that chelation of calcium in egg extracts or specific inactivation of calcium/calmodulin-dependent protein kinase II (CaMKII) blocks even initial centrosome duplication, whereas inactivation of Cdk2 does not. Duplication can be restored to inhibited extracts by addition of CaMKII and calmodulin. These results indicate that calcium, calmodulin, and CaMKII are required for an essential step in initiation of centrosome duplication. Our data suggest that calcium oscillations in the cell cycle may be linked to centrosome duplication.