Repeatability of gaseous measurements across consecutive days in sheep using portable accumulation chambers

Portable accumulation chambers (PACs) enable gaseous emissions from small ruminants to be measured over a 50-min period; to date, however, the repeatability of consecutive days of measurement in the PAC has not been investigated. The objectives of this study were 1) to investigate the repeatability of consecutive days of gaseous measurements in the PAC, 2) to determine the number of days required to achieve precise gaseous measurements, and 3) to develop a prediction equation for gaseous emissions in sheep. A total of 48 ewe lambs (c. 10 to 11 mo of age) were randomly divided into four measurement groups each day, for 17 consecutive days. Gaseous measurements were conducted between 0800 and 1200 hours daily. Animals were removed from perennial ryegrass silage for at least 1 h before measurements in the PAC, and animals were assigned randomly to each of the 12 chambers. Methane (CH₄; ppm) concentration, oxygen (O₂; %), and carbon dioxide (CO₂; %) were measured at three time points (0, 25, and 50 min after entry of the first animal into the first chamber). To quantify the effect of animal and day variation on gaseous emissions, between-animal, between-day, and error variances were calculated for each gaseous measurement using a linear mixed model. The number of days required to gain a certain precision (defined as the 95% confidence interval range) for each gaseous measurement was also calculated. For all three gases, the between-day variance (39% to 40%) accounted for a larger proportion of total variance compared with between-animal variance, while the repeatability of 17 consecutive days of measurement was 0.36, 0.31, and 0.23 for CH₄, CO₂, and O₂, respectively. Correlations between consecutive days of measurement were strong for all three gases; the strongest correlation between day 1 and the remaining days for CH₄, CO₂, and O₂ was 0.71 (days 1 and 6), 0.77 (days 1 and 2), and 0.83 (days 1 and 5), respectively. A high level of precision was achieved when gaseous measurements from PAC were taken over three consecutive days. The prediction equation overestimated gaseous production for all three gases: the correlations between actual and predicted gaseous output ranged from 0.67 to 0.71, with the r² ranging from 0.45 to 0.71. The results from this study will aid the refinement of the protocol for the measurement of gaseous emissions in sheep using the PAC.     

Genetic variability following selection for scrapie resistance in six autochthonous sheep breeds in the province of Bolzano (northern Italy)

Scrapie is an ovine transmissible spongiform encephalopathy, and its susceptibility is associated with polymorphisms in the prion protein gene (PRNP). Genetic selection is currently the most effective mean for eradication of the susceptible VRQ allele in favour of resistant ARR allele. Maintenance of genetic diversity should be one of the major objectives in breeding programmes, especially in endangered breeds, and genetic information are an excellent alternative to pedigree data where these information are missing. The aim of our study was to determine changes of genetic variability in six native sheep breeds from autonomous province of Bolzano, northern Italy, following simulation of scrapie selection scenarios. A total of 684 rams were investigated for PRNP polymorphisms and for 10 microsatellite loci to estimate genetic variability. Across all loci, a total of 163 alleles were detected with a mean of 10.4 alleles per locus. Average observed (Ho) and unbiased expected (uHe) heterozygosity overall loci were 0.74 and 0.78, respectively, showing a statistically significant deviation from Hardy–Weinberg equilibrium (HWE) in all breeds. This heterozygosity deficit was confirmed by a positive fixation index (Fis), determining a moderate inbreeding in each breed. Simulating a soft selection, where only rams having at least a VRQ allele should be excluded from reproduction, Ho, uHe and Fis values remained almost unchanged, indicating that genetic variability should not be affected by the removal of these individuals. With a mild selection scenario, considering only rams with at least one ARR allele, we observed a decrease in the mean alleles per breed (8.9) and the maintenance of heterozygosity deficiency, except for two breeds, where it was any longer significant. These results showed that selection strategies allowing use of heterozygous as well homozygous ARR rams might be the right compromise to improve resistance to scrapie and to do not dramatically affect genetic variability of these breeds.     

Effect of substituting soybean meal with euglena (Euglena gracilis) on methane emission and nitrogen efficiency in sheep

This study evaluated methane (CH₄) emission, intake, digestibility, and nitrogen efficiency in sheep fed diets containing replacement levels (0%, 33%, 50%, and 67% of soybean meal with euglena). In this experiment, four Corriedale wether sheep with an initial body weight of 53.8 ± 4.6 were arranged in a 4 × 4 Latin square design. This experiment lasted 84 days, divided into four experimental periods. Each period lasted 21 days, which consists of 14 days of adaptation to the diets, 5 days to collect samples, and 2 days to collect gas emission from sheep. Methane emission expressed as L/kg DM intake or g/kg DM intake reduced by up to 37% and the energy loss via CH₄ (% of GE intake) reduced by up to 34%. No differences (p > 0.05) were observed in DM and OM intake and whole tract apparent DM digestibility due to substitution of soybean meal with euglena. The total CP loss reduced significantly (linear, p < 0.001) and CP efficiency increased linearly (p = 0.03) with increasing concentration of euglena. As a result, nitrogen balance and average daily weight gain remained unchanged despite higher nitrogen concentration in soybean supplemented group. In conclusion, substitution of soybean meal with euglena reduced methane emission without affecting the performance of animals.     

若尔盖县牦牛及藏系绵羊寄生虫病调查

本文通过剖检20头牦牛和镜检30份新鲜藏系绵羊粪便,对若尔盖县牛羊进行了寄生虫病调查。结果表明:感染牛羊的寄生虫主要为前后盘吸虫、鞭虫、仰口线虫、绦虫和牛蝇蛆,其中前后盘吸虫的危害最大。     

Effect of Terminalia chebula and Allium sativum on in vivo methane emission by sheep

A study was conducted to evaluate some plant parts (already tested for their antimethanogenic activity in in vitro gas production test in the authors’ laboratory) as feed additive to combat methane emission from sheep. Sixteen male sheep with average body weight of 29.96 ± 1.69 kg (22 months of age) were divided into four groups in a randomized block design. The animals were fed on a diet containing forage to concentrate ratio of 1:1. The concentrate fraction composed (in parts) of maize grain, 32; wheat bran, 45; deoiled soybean meal, 20; mineral mixture, 2 and common salt, 1. The four treatments were control (without additive), seed pulp of Terminalia chebula (Harad), bulb of Allium sativum (Garlic) and a mixture (Mix) of the latter two in equal proportions at the rate of 1% of dry matter (DM) intake. There was no effect on DM intake due to the inclusion of these feed additives. The digestibilities of DM and organic matter tended to be higher (p < 0.1) in the groups with T. chebula and A. sativum, whereas, neutral detergent fibre (NDF), acid detergent fibre (ADF) and cellulose digestibilities were higher (p < 0.05) in all the three experimental groups compared with control. The nitrogen balance and plane of nutrition were not affected by inclusion of any of the feed additives. Methane emission (L/kg digested DM intake) as estimated by open circuit respiration chamber and methane energy loss as per cent of digestible energy intake tended to be lower in T. chebula (p = 0.09) and Mix (p = 0.08) groups compared with control. The data indicated that T. chebula showed antimethanogenic activity, whereas both T. chebula and A. sativum improved nutrient digestibility. Therefore, these two plants appear to be suitable candidates for use as feed additive to mitigate methane emission and to improve nutrient utilization by sheep.     

Measurement of progesterone in sheep using a commercial ELISA kit for human plasma

Determination of serum or plasma progesterone (P4) concentrations is important to recognize pregnant and non-pregnant ewes, and also to predict the number of carried lambs. The 2 most common methodologies for the detection of plasma P4 are radioimmunoassay (RIA) and enzyme immunoassay (EIA). RIA is very expensive, and not all laboratories are equipped to perform this test; EIA is commercially available for human use, but only a few companies produce species-specific kits, which are expensive. We verified for ovine plasma a less expensive and easily available ELISA kit (DiaMetra) designed to quantify P4 in humans. Pools of ovine and human plasma were used to compare repeatability, accuracy, sensitivity, and stability of P4 measured by the DiaMetra kit. Repeatability data were within 15%, and accuracy values were ~90% for both plasma matrices. Stability data showed a loss of <20% for freeze–thaw and <30% for 30-d storage. All parameters were acceptable under international guidelines for method validation. The human ELISA kit was used successfully to quantify plasma P4 in 26 ewes during pregnancy until delivery. P4 concentrations were also correlated with the number of carried lambs.     

河北省不同地区夏季规模化羊场气载细菌的检测与分析

旨在研究河北省夏季不同地区规模化羊场气载细菌的日变化和区域性变化。选择3个区域(燕山丘陵、太行山区和平原地区)10个规模化羊场(14个舍),采用定点采样和培养计数法,对夏季各羊舍及其运动场的细菌数量进行3 d连续检测。结果:羊舍和运动场细菌数量表现为中午低、早或晚高的变化规律,早、中、晚不同地区羊舍的细菌数量分别为6.23×10^3~9.78×10^3、5.05×10^3~6.56×10^3和6.05×10^3~7.51×10^3 cfu/m^3,其中太行山区羊舍细菌在3个时间段间达显著差异(P<0.05)。从区域性分布看,太行山区羊舍细菌数量高于其他2个地区,燕山、太行和平原3个地区羊舍细菌平均数量分别达6.76×10^3、7.79×10^3和6.01×10^3 cfu/m^3;羊舍与运动场比较,虽然太行山区和平原地区羊舍细菌与运动场之间无显著性差异(P>0.05),但太行山区羊舍及其运动场的细菌数量有增加趋势(P=0.05,P=0.07),而燕山丘陵50%的羊舍细菌数量显著高于运动场(P<0.05)。另外,比较4类羊(妊娠、母带仔、育成和育肥)舍细菌得出,妊娠舍细菌平均数量最高(7.10×10^3 cfu/m^3),育肥舍或带仔母羊舍细菌数量最低(5.54×10^3 cfu/m^3,5.55×10^3 cfu/m^3)。该研究结果可为完善羊场的羊舍设计和环境调控提供可靠依据。     

Morphometric study of the testis in sheep embryos using unbiased design-based stereology

Sheep have been used as translational models of human postnatal testicular development. However, the morphometric features of the normal developing testis in sheep embryos have not been previously investigated using stereology. The objective of the present work was to establish normal quantitative parameters for fetal testicular tissue components in sheep, using unbiased design-based stereological methods. Twenty-four sheep embryos were divided into four gestational age groups (9–11, 12–14, 15–17 and 18–20 weeks of gestation) on the basis of the embryos’ crown-rump length. Isotropic, systematic uniform random sections of the left testes were obtained by employing the orientator method. Testicular total volume, the absolute and proportional volumes occupied by the seminiferous tubules and interstitial tissue, as well as the seminiferous tubule length, were estimated using the point-counting system and the unbiased counting frame principle. All the parameters, with the exception of the interstitial tissue's fractional volume, gradually increased along with gestational age, with the maximum increase especially seen in the late fetal stages. The proportional volume of the interstitial tissue, on the other hand, showed a decreasing trend along with increasing gestational age. The absolute volume of the testes, of the seminiferous tubules and of the interstitial tissue, and the length of the seminiferous tubules showed a significant (p< 0.05) positive linear correlation with gestational age. Several similarities were observed with human testicular embryogenesis. The stereological data emerging from the present study might prove useful as basic contribution to the fields of andrology and embryology and stimulate further research in these areas.     

Analysis of genetic variability in the Czech Dachshund population using microsatellite markers

The primary goal of this study was to analyse genetic variation within and among six Dachshund varieties in the Czech Republic using 10 microsatellites from StockMarks^® Paternity Polymerase Chain Reaction (PCR) Typing kit. A total of 632 Dachshunds were sampled – 99 Standard Smooth-haired, 72 Standard Long-haired, 272 Standard Wire-haired, 42 Miniature Smooth-haired, 73 Miniature Long-haired and 74 Miniature Wire-haired. Average observed heterozygosity and polymorphic information content ranged in particular Dachshund varieties between 0.58–0.70 and 0.57–0.64, respectively. Subsequent analysis of the population structure ( F -statistics; Nei's genetic identity) showed that Standard Dachshunds shared allele frequencies most closely with their miniature equivalents, and smooth coat type is closer to Wire-haired coat type than to the Long-haired one. Analysis of molecular variance revealed that 11.81% of the total variance occured among varieties. The value of combined exclusion probability (0.9955) validated usefulness of this panel of microsatellites for parentage verification in all analysed populations. We carried out 234 parentage verifications with 1.28% cases where parentage did not match.     

宁夏羊胃肠道线虫对现行驱虫药的抗药性调查

采用粪便虫卵减少试验对宁夏地区所属灵武、贺兰、盐池、吴忠、中宁、中卫、永宁和银川市郊8个县（市）的12个绵羊场、6个山羊场进行了丙硫苯咪唑和阿维菌素抗药性的随机调查。结果表明：在用丙硫苯咪唑调查的10个绵羊场和6个山羊场中，虫卵减少率在95％以下和95％的置信域下限在90％以下的有山羊场2个、绵羊场2个，证明对丙硫苯咪唑有抗药性；1个绵羊场和1个山羊场的虫卵减少率是96．3％、95．9％，但95％置信域的下限在90％以下，具有抗药性可疑；山羊群中丙硫苯咪唑的抗药性为33．3％，绵羊群为20．0％。用同样的方法调查了1个山羊场和5个绵羊场（其中有4个羊场曾执行了丙硫苯咪唑的试验），查出1个山羊场和1个绵羊场对阿维菌素具有抗药性可疑，其虫卵减少率分别为97．3％和95．5％，置信域下限在90％以下。揭示了宁夏地区羊消化道线虫对现行驱虫药的抗药状态，为今后防治提供了依据。     

Heart rate variability parameters as markers of the adaptation to a sealed environment (a hypoxic normobaric chamber) in the horse

Simulated hypoxic normobaric devices have been used in human beings in order to enhance endurance capacity. These devices are sealed chambers where the athletes are supposed to stay for at least 6–8 hr daily. The current research assesses the changes in time‐domain, spectral and non‐geometrical heart rate variability (HRV) parameters in 6 horses subjected to progressive duration periods inside of a hermetically sealed chamber. It was pursued, firstly to evaluate the intensity of the stress experienced by the animals and secondly to elucidate whether the horses might require an acclimation period before implementation of hypoxic conditions. HRV parameters were monitored for 6 days: day 0 (6‐hr duration; in paddocks; basal conditions), and days 1, 2, 3, 4 and 5 (1, 2, 3, 4 and 6 hr inside the chamber every day respectively). During day 1 and during the first hours of days 2 and 3, compared to day 0, horses presented increased HR and SDHR values and decreased RR interval duration. SD1 values decreased on some hours of days 2 and 3, but differences with day 0 were not found on day 1. Increased SDNN, RMSSD, SD1 and SD2 values were observed on days 4 and 5. These results showed an activation of the sympathetic activity together with an attenuation of the parasympathetic activity during the days 1 to 3. Increased parasympathetic activity was found only during the first hours of days 4 and 5. Spectral parameters experienced minor variations, with increased LF_peak and LF% during some hours of days 4 and 5. In conclusion, at least 3 days are needed to adapt the horse to a sealed environment before starting to subject the animals to hypoxic conditions. When the horses were acclimatized, however, a minor stress was detected with they spent more than 4 hr inside of the chamber.     

武威市小规模牛羊场粪污资源化利用现状调研与分析

通过入户调研武威市三县一区牛羊规模养殖整体情况和不同规模牛羊场资源化利用现状,针对小规模牛羊场量多,治理难度大,治理工艺不完善,粪污资源化利用收转运体系不健全等问题,分析小规模牛羊场如何因地制宜进行粪污资源化利用。     

Nutritional evaluation of the legume Macrotyloma axillare using in vitro and in vivo bioassays in sheep

This study consisted of two experiments with the following objectives: to evaluate the effects of tannins from the tropical legume macrotiloma (Macrotyloma axillare) on total gas and methane (CH₄) production, as well as on ruminal fermentation parameters by performing an in vitro bioassay, with samples incubated with and without polyethylene glycol (PEG) in a semi‐automatic system; and secondly in a 17 day in vivo experiment, to determine apparent total tract digestibility (ATTD) of dietary nutrients and ruminal fermentation parameters of 12 intact 8‐ to 9‐month‐old Santa Inês (averaging 24.95 ± 1.8 kg body weight) ewes fed tropical grass hay supplemented with macrotiloma hay. The ewes were divided into two treatment groups depending on their diet: chopped aruana grass hay (Panicum maximum cv. Aruana) (control—CON); and aruana grass hay supplemented with chopped macrotiloma hay (macrotiloma—MAC). The animals were kept for 5 consecutive days in metabolic cages for the ATTD assay, and at the end of this period, samples of rumen fluid were collected from each ewe to determine ammoniacal nitrogen (NH₃‐N) and short‐chain fatty acid (SCFA) production, and protozoa count. For the in vitro assay, a decrease in total gas and CH₄ production was observed for samples incubated without PEG (p < .05). No differences were observed for the other parameters evaluated (p > .05). In the in vivo experiment, increased intake and ATTD of crude protein were observed for the animals fed MAC when compared to CON (p < .05). For rumen fermentation parameters, increased NH₃‐N, total SCFA and isobutyrate concentrations, as well as reduced protozoa count were observed for MAC when compared to CON (p < .05). The results observed here indicated the potential of macrotiloma for use as a ruminant feed, and antimethanogenic potential of this plant was noted.     

我国部分地区羊消化道寄生虫感染情况调查及球虫种类鉴定

为了解我国部分地区羊肠道寄生虫的感染情况,采用水洗离心沉淀法、卢戈氏碘液染色法和饱和蔗糖溶液漂浮法,对来自全国8省16个地区共472份羊粪样品进行了肠道寄生虫的显微镜检查,并根据虫卵和卵囊形态进行了虫种鉴定。本次调查寄生虫总感染率为85.38%(403/472),混合感染率为46.40%(219/472),共查到球虫、阿米巴、线虫和绦虫4大类寄生虫,感染率分别为68.86%(325/472)、44.49%(210/472)、29.24%(138/472)和1.91%(9/472)。球虫种类鉴定结果显示共10种,分别为阿氏艾美耳球虫(Eimeri arloingi)、艾丽艾美耳球虫(Eimeri alijevi)、苍白艾美耳球虫(Eimeri pallida)、家山羊艾美耳球虫(Eimeri hirci)、山羊艾美耳球虫(Eimeri caprina)、阿撒他艾美耳球虫(Eimeri ahsata)、斑点艾美耳球虫(Eimeri punctata)、羊艾美耳球虫(Eimeri caprovina)、颗粒艾美耳球虫(Eimeri granulosa)和克氏艾美耳球虫(Eimeri chri...     

Surgical anatomy of the nasal and paranasal sinuses in Egyptian native sheep (Ovis aries) using computed tomography and cross sectioning

The present work aimed to describe the normal computed tomography (CT) and cross‐sectional anatomy of the nasal and paranasal sinuses in sheep and to correlate these features with the relevant clinical practices. Twenty apparent healthy heads of Egyptian native breed of sheep (Baladi sheep) of both sexes were used for studying these sinuses. CT images and their closely identical cross sections of the same head were selected and serially labelled in a progression from the rostral nasal region to the caudal aspect of the head using cheek teeth as landmarks. The current investigation reported seven sinuses in sheep, including maxillary, frontal, lacrimal and sphenoidal as paranasal, as well as dorsal and middle conchal and ethmoidal as nasal with unnoticeable palatine and ventral nasal conchal sinuses. The boundaries, extension, structure and communications of these sinuses were fully described. The current study provided anatomical guidelines for surgical interference in the frontal and maxillary sinuses during trephination, dehorning and sinuscopy. Also, an acceptable anatomical explanation was reported in this study for the high incidence of maxillary sinusitis than other sinuses. CT and cross‐sectional anatomy could be used as helpful database for diagnosis and clinical interference of the nasal and paranasal sinuses in sheep.     

Thoracic epidural analgesia via the lumbosacral approach using multiport catheters with a low concentration of bupivacaine and morphine in sheep

ObjectiveTo determine the analgesic and systemic effects of thoracic epidural administration of bupivacaine (BP) and morphine (MP) in conscious sheep.Study designRandomized, crossover, experimental study.AnimalsSix healthy castrated sheep weighing between 40 and 50 kg.MethodsEach sheep received, via the lumbosacral approach, BP (0.5 mg kg^?1), MP (0.1 mg kg^?1), and BP plus MP (BPMP; 0.25 mg kg^?1 + 0.05 mg kg^?1) in a randomized order. Heart rate, blood pressure, respiratory rate, blood gas analysis, skin temperature, rectal temperature, analgesia, sedation, and motor blockade were determined before treatment and at predetermined intervals until analgesia had disappeared.ResultsThe main areas of complete analgesia for the BP and BPMP treatments were the thorax and forelimb bilaterally. The median duration of analgesia was shorter with MP treatment (45 minutes; score 2) than with BP treatment (70 minutes) and BPMP treatment (140 minutes; p < 0.05). The BP and BPMP treatments caused motor block, and MP and BPMP treatments showed mild sedation. Significant decreases in systolic and diastolic arterial blood pressures were observed only with the BP treatment (p < 0.05). Epidural MP combined with the BP local anesthetic depressed ventilation but within acceptable limits in these clinically healthy sheep.ConclusionsThoracic epidural administration of BPMP to sheep resulted in longer duration of analgesia of the thorax and forelimbs bilaterally in conscious sheep than the administration of MP or BP alone. The incidence of complications was low, but side-effects such as depressed ventilation and muscle paralysis occurred and require appropriate management.Clinical relevanceThis technique should be considered as another method for the relief of postoperative pain after thoracic surgery in sheep.     

陕南贫困地区应用微生态制剂提高肉羊养殖经济效益研究

本文主要从肉羊养殖角度来对陕南贫困地区应用微生态制剂的情况做出分析,希望可以进一步提高肉羊养殖的经济效益。试验选取体重一致的90d肉羊240只,随机分为3组,每组10个重复,每个重复8只羊。试验共设计3种日粮:对照组饲喂未加工的玉米秸秆日粮,试验1组饲喂未加工的青贮玉米秸秆日粮,试验2组饲喂添加EM菌剂发酵后的玉米秸秆。试验共开展2个月。结果发现,采用EM菌剂发酵玉米秸秆饲喂的肉羊体重增加6.71kg/只,增重及盈利均明显高于试验I组及对照组,收益分别提高了16.99元/只和4.87元/只。因此,微生态制剂能提升肉羊养殖的经济效益,可予以推广应用。     

Comparison of repeatability and multiple trait threshold models for litter size in sheep using observed and simulated data in Bayesian analyses

Bayesian analyses were used to estimate genetic parameters on 5580 records of litter size in the first four parities from 1758 Mule ewes. To examine the appropriateness of fitting repeatability (RM) or multiple trait threshold models (MTM) to litter size of different parities, both models were used to estimate genetic parameters on the observed data and were thereafter compared in a simulation study. Posterior means of the heritabilities of litter size in different parities using a MTM ranged from 0.12 to 0.18 and were higher than the heritability based on the RM (0.08). Posterior means of the genetic correlations between litter sizes of different parities were positive and ranged from 0.24 to 0.71. Data sets were simulated based on the same pedigree structure and genetic parameters of the Mule ewe population obtained from both models. The simulation showed that the relative loss in accuracy and increase in mean squared error (MSE) was substantially higher when using the RM, given that the parameters estimated from the observed data using the opposite model are the true parameters. In contrast, Bayesian information criterion (BIC) selected the RM as most appropriate model given the data because of substantial penalty for the higher number of parameters to be estimated in the MTM model. In conclusion, when the relative change in accuracy and MSE is of main interest for estimation of breeding values of litter size of different parities, the MTM is recommended for the given population. When reduction in risk of using the wrong model is the main aim, the BIC suggest that the RM is the most appropriate model.     

用丙硫苯咪唑检测羊线虫抗药性的体外虫卵孵化试验

为探讨用丙硫苯咪唑进行体外虫卵孵化试验检测羊线虫对苯并咪唑类药物的抗药性,对23个羊场的25份田间样品用丙硫苯咪唑和噻苯咪唑进行了虫卵孵化试验并与先前减卵试验（faecal egg count re-duction test,FECRT）的结果比较。结果表明,25份样品中,丙硫苯咪唑和噻苯咪唑对受检虫卵的半数致死量（LD50）均值分别为0.050 1和0.054 0μg/mL,差异不显著。FECRT检测有抗药性的4个羊场,75%的样品对丙硫苯咪唑和噻苯咪唑的LD50均值均在0.1μg/mL以上,只有1个羊场的的LD50值分别为0.068 9μg/mL（丙硫苯咪唑）和0.071 2μg/mL（噻苯咪唑）。检测为可疑的2个羊场,其样品的LD50值为0.04～0.07μg/mL;FECRT检测敏感的蠕虫群体的LD50均在0.04μg/mL以下。此外,丙硫苯咪唑用纯的二甲基亚砜溶解和稀释后于4℃保存7 d,LD50值变化不大,提示药效无明显下降,而保存14 d后药效有下降趋势。