Stallion Semen Cooling Using Native Phosphocaseinate-based Extender and Sodium Caseinate Cholesterol-loaded Cyclodextrin-based Extender

The objective of this study was to compare semen parameters and embryo recovery rates of cooled stallion semen extended with INRA 96 or BotuSemen Gold. In experiment 1, 45 ejaculates from nine mature stallions were collected, assessed, and equally split between both extenders and then extended to 50 million sperm/mL. Then, the extended semen was stored in three passive cooling containers (Equitainer, Equine Express II, and BotuFlex) for 48 hours. In experiment 2, the same ejaculates extended in experiment 1 were cushion-centrifuged, the supernatant was discarded, and the pellets were resuspended at 100 million sperm/mL with their respective extender. Semen was then cooled and stored as in experiment 1. In both experiments, sperm motility parameters, plasma membrane integrity, and high mitochondrial membrane potential were assessed at 0, 24, and 48 hours post cooling. For experiment 3, 12 mares (n = 24 cycles) were bred with 48 hour–cooled semen from one stallion. Semen was processed as described in experiment 1. Mares had embryo flushing performed by 8-day post-ovulation. In experiment 1, BotuSemen Gold displayed superior total and progressive motility relative to INRA 96 (P < .05). There were no significant differences between the types of containers in any experiment. In experiment 2, INRA 96 and BotuSemen Gold extenders had similar total and progressive motility, but BotuSemen Gold had superior sperm velocity parameters at all timepoints. Embryo recovery was identical for both extenders (50%). Finally, the results obtained herein suggest that BotuSemen Gold is a suitable alternative to be included in semen cooling tests against INRA 96 in clinical practice.     

Evaluation of Brix Refractometry for the Estimation of Colostrum Quality in Jennies

Donkey placenta does not allow the passage of immunoglobulins; thus, foals are born hypogammaglobulinemic and an adequate intake of high-quality colostrum in the first 24 hours of life is crucial for the surviving. The study aims to assess the relation between colostrum immunoglobulin G (IgG) concentration evaluated by the single radial immunodiffusion (SRID) test and the Brix refractometer in donkeys to establish a cutoff value for high quality of colostrum based on Brix refractometry. Colostrum was collected at foaling, and at 6, 12, and 24 hours after foaling from the left and the right half of nine Amiata jennies. A total of 72 colostrum samples were analyzed. A Friedman test with a Dunn’s test for multiple comparisons was used for assessing the differences between the left and right half at each sampling time. No differences were found between the left and right halves; the average value was used to analyze the effect of sampling time on the IgG concentrations and Brix values. The relationship between colostrum IgG concentrations (SRID test) versus Brix value and Brix value versus time was analyzed using two different mixed linear models. A strong statistically significant relation has been found between IgG concentrations and Brix values (R² = 0.84). The relation between IgG concentrations and Brix refractometer showed a cutoff point of 17% Brix score for the identification of high-quality colostrum. The Brix value (%) decreased continuously from 16.29 by 0.29 × hour. Jennies’ and donkey foals’ management may be greatly improved using this simple and cheap device.     

雏鸡中枢免疫器官内B淋巴细胞变化规律

为了研究雏鸡在正常发育过程中免疫器官内B淋巴细胞的变化规律,以不同日龄的SPF鸡为研究对象,用流式细胞术检测鸡脾脏、胸腺及法氏囊内IgA+、IgG+及IgM+B淋巴细胞含量的变化情况。结果显示,在脾脏中,4日龄时B淋巴细胞含量比较多,7日龄时B淋巴细胞含量下降,随后,7至21日龄细胞含量逐渐升高;胸腺内则是在4日龄时IgM+B淋巴细胞含量相对较高,而IgA+、IgG+B淋巴细胞几乎没有;在法氏囊中,IgA+、IgG+及IgM+B淋巴细胞含量在4至14日龄时逐渐升高,并且IgA+和IgG+细胞含量在21日龄时基本趋于稳定,但是IgM+B细胞含量则在21日龄时有所下降。试验结果表明,雏鸡出壳后各免疫器官B淋巴细胞始终以IgM+细胞含量最多。     

蜂王浆对尾静脉注射4T1细胞小鼠抗氧化能力的影响

探讨蜂王浆对尾静脉注射4T1细胞小鼠的抗氧化和免疫力的影响.将4T1细胞或其与蜂王浆的混合物通过尾静脉注射入小鼠体内,ELISA法检测蜂王浆对小鼠血清、肝脏和肾脏中与抗氧化能力相关的指标及血清中和免疫力相关细胞因子水平的影响.结果表明,和模型对照组相比,蜂王浆(50、100 mg·mL~(-1))可以显著提高小鼠血清内T-AOC、IFN-α(P0.05)水平,降低IL-4的水平(P0.05);蜂王浆剂量为50 mg·m L~(-1)时,IFN-γ的含量显著增加(P0.05).蜂王浆(100 mg·mL~(-1))显著提高肾脏中T-AOC的水平(P0.05).可见,蜂王浆可以改善尾静脉注射4T1细胞小鼠的抗氧化能力和免疫力.     

Effect of anaesthesia on cell-mediated immunity in dogs undergoing mastectomy for mammary cancer

ObjectiveTo investigate if anaesthesia for canine cancer mastectomy further influences host cell-mediated immunity (CMI) promoting cancer progression.Study designA randomized, controlled, blinded clinical study.AnimalsA total of 20 bitches with malignant mammary tumours of clinical stage II or III undergoing the same type of mastectomy (regional mastectomy).MethodsDogs were randomly allocated to one of two anaesthetic groups (10 per group). The anaesthetic protocol of group A used minimally immunosuppressive drugs (tramadol, robenacoxib, propofol), whereas that of group B (control) used more immunosuppressive drugs (morphine, fentanyl, thiopental, isoflurane). For each animal, measurements of white blood cells (WBCs), neutrophils and lymphocytes, and flow cytometric assessment of T cells (CD3⁺), helper T cells (CD4⁺), cytotoxic T cells (CD8⁺) and CD5^low+ T cells were performed prior to anaesthesia (day 0) and on days 3 and 10 postsurgery. Data were analysed using a General Linear Model for repeated measures and presented as mean ± standard deviation, p ≤ 0.05.ResultsIn all animals, on day 3, WBCs and neutrophils were significantly increased (p < 0.0005), while flow cytometry revealed significantly decreased relative percentages of T cells (CD3⁺) (p = 0.003) and their subpopulations CD4⁺ (p = 0.006), CD8⁺ (p = 0.029) and CD5^low+ (p = 0.031). Specifically, on day 3, the cytotoxic T cells (CD8⁺) were significantly decreased (p = 0.05) only in group B, whereas the CD4⁺ (p = 0.006) and CD5^low+ (p = 0.008) T cells in group A. The only significant difference between groups was found preoperatively in the CD4⁺/CD8⁺ ratio, which was higher in group A (p = 0.006).Conclusions and clinical relevanceIn dogs with mammary cancer undergoing regional mastectomy, a significant decrease in components of CMI was observed on day 3 postsurgery in both anaesthetic groups. Some indication, however, for better preserved cellular immunity by less immunosuppressive anaesthetic/analgesic drugs was detected, rendering their use advisable.     

β-羟丁酸抑制脂多糖诱导奶牛中性粒细胞核因子-κB信号通路的激活

高酮血症造成奶牛中性粒细胞先天免疫机能受到抑制,本研究探讨β-羟丁酸(BHBA)是否抑制脂多糖(LPS)诱导的奶牛中性粒细胞核因子-κB(NF-κB)信号通路的激活。分离健康奶牛中性粒细胞,采用LPS(100 ng/mL)和不同浓度(0.5、1.0、2.0和4.0 mmol/L)BHBA作用于中性粒细胞,收集细胞,应用实时荧光定量PCR(qRT-PCR)检测中性粒细胞中白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)和NF-κBp65 mRNA表达水平,Western blot检测NF-κBp65蛋白表达水平,比色法检测核因子-κB抑制物激酶β(IKKβ)激酶活性,酶联免疫吸附试验(ELISA)法检测促炎细胞因子TNF-α、IL-6和IL-1β的分泌量。结果表明:与对照组(不进行BHBA和LPS处理)相比较,LPS组(单独LPS处理)中IL-1β、IL-6、TNF-α和NF-κBp65 mRNA表达水平和NF-κBp65蛋白表达水平极显著增加(P <0.01),IKKβ激酶活性极显著增强(P<0.01),IL-1β和TNF-α的分泌量极显著增加(P<0.01),IL-6的分泌量显著增加(P <0.05)。与LPS组相比,0.5、1.0、2.0和4.0 mmol/L BHBA均使IL-1β、IL-6、TNF-α和NF-κBp65 mRNA表达水平和NF-κBp65蛋白表达水平显著或极显著增加(P<0.05或P<0.01),IKKβ激酶活性显著或极显著增强(P<0.05或P<0.01),IL-1β、IL-6和TNF-α的分泌量显著或极显著增加(P<0.05或P<0.01)。以上结果表明,高浓度BHBA可以抑制LPS诱导的奶牛中性粒细胞NF-κB信号通路的激活,具有一定的抗炎功能。     

复方中草药饲喂时间对凡纳滨对虾消化和免疫酶活性的影响

配制4组饲料,饲料中复方中草药的添加量分别为0%(对照组)、0.1%、0.2%和0.4%,每组3个重复,连续投喂凡纳滨对虾21 d,每7 d采样检测肝脏和肠道消化酶及血清免疫酶活性的变化。实验结果表明,实验组消化酶活性基本都高于对照组,随着时间的变化呈波动趋势,0.4%组的肝蛋白酶和肠蛋白酶第7 d时最高(P<0.05);肝淀粉酶指标出现逐渐下降的趋势,而肠淀粉酶呈逐渐上升的趋势,0.2%组的肠淀粉酶活性在第7 d显著增高(P<0.05)。0.2%组的超氧化物歧化酶(SOD)在第14 d最高(P<0.05);过氧化物酶(POD)指标变化不明显。因此,阶段性投喂中草药对凡纳滨对虾消化酶和免疫酶有一定的促进作用,能提高对虾的消化和免疫能力。     

蜂胶对小鼠T细胞亚群及主要相关细胞因子分泌的影响

为阐明蜂胶对小鼠特异性细胞免疫应答的影响特点,选择1、2、4 mg/ 3个剂量分别经腹腔注射给6周龄Balb/c小鼠,在0、7、10、15 d采血样,采用流式细胞术、ELISA分别测定小鼠外周血CD4+T细胞、CD8+T细胞数量、血清中IFN-γ、IL-4含量.结果显示,与注射蜂胶前相比,各不同剂量的蜂胶均能够使CD4...     

Serum complement activity in the three-toed amphiuma (Amphiuma tridactylum)

Some animals routinely endure serious injuries from predators or during intraspecific territorial conflicts. Such is the case for Amphiuma tridactylum, an aquatic salamander that lives in an environment rich in potentially infectious microbes, apparently with rare or no pathogenic infection. Some vertebrates possess innate immune mechanisms, but whether this is the case for Amphiuma is unknown. To assess this potential, plasma from 19 A. tridactylum was pooled and used for characterisation of serum complement activity. The ability of A. tridactylum plasma to hemolyse unsensitised sheep red blood cells (SRBCs) was titer-dependent, with low activity observed even at high plasma titers. The kinetic characterisation of SRBC hemolysis revealed that significant activity could be measured within 10 min of incubation, and maximal activity occurred within 60 min. The SRBC hemolysis by A. tridactylum plasma was also temperature-dependent, with maximal activity at 30 °C. In addition, this activity was sensitive to mild heat treatment, with 96% of activity inhibited by incubation at 56 °C for 30 min. The SRBC hemolysis could also be inactivated by pretreatment of the plasma with proteases, indicating that this activity was protein dependent. The activity required divalent metals ions, with activity inhibited by EDTA, citrate, or phosphate. However, the chelator-inhibited activity could be restored by the addition of excess Ca²⁺ or Mg²⁺, but not Cu²⁺ or Ba²⁺, indicating specificity of the divalent metal ion requirement. The sensitivity to heat, proteases, and divalent metal ion chelators strongly suggests that A. tridactylum plasma-mediated hemolysis of SRBCs is mediated by the serum complement system of proteins.     

辣椒连作研究进展

辣椒是一种重要蔬菜作物,由于连年耕种辣椒存在连作障碍问题;国内外学者对辣椒连作问题作了较深入的研究.针对国内外辣椒连作的研究进展,主要从土壤微生物环境、连作土壤的微生态条件、辣椒自毒分析和化感作用、辣椒免疫系统及自身抗性作用机理等方面分别进行了综述,并对其今后的发展方向作了展望.