Serum biochemical parameters and embryo production during superovulatory treatment in dairy cattle

The objective of this study was to determine the relationship between the number of transferable embryos (TE) and various blood chemistry parameters as a reflection of the metabolic state of cows after superovulatory treatment. Forty-nine Holstein cows were subjected to superovulatory treatment for commercial embryo production. At the time of embryo harvest, individual blood samples were taken from cows for biochemical analysis. All embryos including dead ones as well as non-fertilized oocytes were counted in uterine lavage. Feed samples collected daily for a period of two weeks before embryo harvest, were analyzed for mycotoxins: vomitoxin, zearalenone and T-2 toxin. On average, cows produced 9.45 ± 5.60 embryos and oocytes of which 5.27 ± 4.20 were TE, 0.37 ± 0.80 were dead embryos and 3.82 ± 3.78 were non-fertilized oocytes. Higher concentrations of Mg and K were associated with a higher production of TE (p = 0.005 and p = 0.043, respectively) and higher activity of creatinine kinase was associated with a lower production of TE (p = 0.011).     

Evaluation of immunochromatographic assay for serodiagnosis of Brucella canis

Canine brucellosis is a contagious disease with venereal and oral modes of transmission that produces late abortion in females, epididymides and prostates in males. Diagnosis is difficult because of unstable serum antibody titers that vary from individual to individual as well as between different methods used for their detection. The objective of this work was to evaluate the clinical utility of the immunochromatographic assay (ICA) for serodiagnosis of dogs suspected of having brucellosis, and results were compared with those obtained for hemoculture (HC) and the rapid screening agglutination with 2-mercaptoethanol (2-ME RSAT). The all experimentally infected dogs were positive in ICA, HC and 2-ME RSAT from 5 weeks, 7 weeks, and 3 weeks after infection, respectively. Also, among dogs selected from 10 different breed kennels occurred brucellosis, 24.8%, 39.5% and 39.1% of dogs tested were detected as positive with HC, 2-ME RSAT and ICA, respectively. The kappa value between 2-ME RSAT and ICA was 0.89. The results of this study showed that sensitivity and specificity of the ICA are comparable with those obtained using conventional serological and bacteriological test for brucellosis. In conclusion, the ICA kit provides a handy and accurate tool for the rapid serodiagnosis of canine brucellosis.     

Cloning, expression and bioassay of canine CTLA4Ig

Blockade of the B7:CD28 costimulatory pathway has been shown to inhibit humoral immunity, graft rejection, graft versus host disease and ameliorate autoimmune diseases. A soluble chimeric fusion protein, CTLA4Ig, binds to B7 with greater affinity than CD28 and blocks the binding of CD28 to B7. We describe the cloning and expression of canine CTLA4Ig, a recombinant chimeric fusion protein composed of the extracellular domain of canine CTLA-4 and the CH2-CH3 domains of canine immunoglobulin alpha constant region (IGHA) genes, linked via an immunologically inert flexible peptide. The recombinant CTLA4Ig protein of approximately 45kDa molecular weight was expressed mainly as insoluble inclusion bodies in Escherichia coli. The protein was solubilized in denaturing buffer and purified using nickel-nitrilotriacetic acid (Ni-NTA) affinity column chromatography followed by refolding. The yield was about 6mg of recombinant CTLA4Ig per liter of culture. The purified protein was biologically active in one-way mixed lymphocyte reactions, demonstrating immunosuppressive activities in a dose-dependent manner. The findings suggest that recombinant canine CTLA4Ig protein could be valuable in assessing the function of CTLA-4 in the canine immune system and may be effective in autoimmune disease therapy.     

Immunolocalization of insulin-like growth factor-I (IGF-I) and its receptors (IGF-IR) in the equine epididymis

Insulin-like growth factor plays a paracrine/autocrine role in regulating testicular function in the stallion, but its presence in the equine epididymis remains unknown. The aim of this study was to test the hypothesis that insulin-like growth factor-I (IGF-I) and IGF-I receptor (IGF-IR) are localized in the caput, corpus, and cauda of the epididymis in an age-dependent manner. Immediately after castration, epididymal tissue was fixed, paraffin-embedded, and processed for immunohistochemistry (IHC). Western blot was also performed using equine epididymal extracts to verify the specificity of the antibodies against IGF-I and IGF-IR. Immunolabeling of IGF-I was observed in the cytoplasm of principal and basal cells in the caput, corpus, and cauda at the pre-pubertal (3–7 months), pubertal (12–18 months), post-pubertal (2–4 years), and adult stages (4.5–8 years). Immunolabeling of IGF-IR was observed in the cytoplasm of principal cells in all regions of the epididymis in each age group. Immunolabeling of IGF-IR was also detected in the cytoplasm of basal cells from animals of all ages. Bands observed by Western blot corresponded to the molecular weights of IGF-I and IGF-IR, ~23 kDa and 95 kDa, respectively. These results suggest that IGF-I might function as an autocrine and/or paracrine factor during the development, maintenance and/or secretions of the stallion epididymis.     

Immunological study of an attenuated Salmonella Typhimurium expressing ApxIA,ApxIIA, ApxIIIA and OmpA of Actinobacillus pleuropneumoniae in a mouse model

Salmonella Typhimurium strain expressing the Actinobacillus pleuropneumoniae antigens, ApxIA, ApxIIA, ApxIIIA and OmpA, was previously constructed as a vaccine candidate for porcine pleuropneumonia. This strain was a live attenuated (∆lon∆cpxR∆asd)Salmonella as a delivery host and contained a vector containing asd. An immunological study of lymphocyte proliferation, T-lymphocyte subsets and cytokines in the splenocytes of a mouse model was carried out after stimulation with the candidate Salmonella Typhimurium by intranasal inoculation. The splenic lymphocyte proliferation and the levels of IL-4, IL-6 and IL-12 of the inoculated mice were significantly increased, and the T- and B-cell populations were also elevated. Collectively, the candidate may efficiently induce the Th1- and Th2-type immune responses.     

The influences of weaning age and weight on carcass traits and meat quality of pigs

The purpose of this study was to establish the effects of weaning age and weight on pigs, and their interaction with carcass traits and meat quality. A total of 468 piglets were obtained from 57 sows and four boars and grouped by age at weaning (D21, 18–24 days; D28, 25–32 days). Each weaning group was subdivided into three weight groups (L, M and H) according to weaning weight. The D28 group had heavier carcass weight, redness and yellowness, but had lower marbling scores and less drip loss than the D21 group (P < 0.05). The pigs with a light weight at weaning had higher carcass weights and lower yellowness than did pigs with a medium or heavy weight at weaning (P < 0.05). Weaning age was found to have a negative correlation with drip loss, while weaning weight was negatively correlated with carcass weight and drip loss (P < 0.05). We concluded that carcass and meat quality traits in pigs were significantly related to their age and weight at weaning. Therefore, we find that piglet weaning age and weight are no less important than post‐weaning growth performance and behavior, with regard to carcass traits and meat quality.     

Immune responses to oral vaccination with Salmonella-delivered avian pathogenic Escherichia coli antigens and protective efficacy against colibacillosis

In this study, the immune responses to and protective efficacy of a live attenuated Salmonella-delivered vaccine candidate secreting the papA, papG, iutA, and clpG antigens of Escherichia coli were evaluated against infection with avian pathogenic E. coli (APEC) in layer chickens. Primary vaccination was done at age 7 d and booster vaccination at age 5 wk. The levels of intestinal secretory immunoglobulin A specific to the 4 antigens were significantly higher in the vaccinated group than in the control group. A potent lymphocyte-proliferation response and increased levels of interferon-γ, interleukin-2, and interleukin-6 in the plasma and in culture supernatants of antigen-stimulated lymphocytes from the vaccinated group suggested significant induction of the cell-mediated immune response in this group compared with the control group. Upon challenge with a virulent APEC strain at 8 wk of age, the vaccinated group had no deaths, whereas the control group had a 15% mortality rate. In addition, the morbidity rate was significantly higher in the control group (55%) than in the vaccinated group (15%). Thus, giving primary and booster vaccination with the Salmonella-delivered APEC vaccine candidate significantly elevated both mucosal and cellular immune responses, which protected the chickens against colibacillosis.     

猪苓的人工培养条件研究

猪苓的白色长菌丝在PDA培养基上能够形成厚实可剥的菌丝块。猪苓生长菌丝可形成锁状联结、顶端膨大,菌丝真径0.95～2.80um,适宜生长条件为20℃、pH4。振荡培养下,形成15～20um大小的菌丝块,颜色如潜伏其菌核的灰黄色。在6种生长原木上,猪苓形成菌丝块大小为3～5um。在PDA培养基上猪苓与蜜环菌混合培养比单独培养的菌丝粗,菌丝密度大,在原木上,猪苓菌丝与蜜环菌菌丝形成共生联结,同时在与蜜环菌紧连的菌丝块边缘,有多点状的自我防御结构。木质素降解活性检测表明,4个菌株中的A、B、C使澳酚蓝脱色后的吸光度为0.0635～0.0902,而菌株D的活性可以忽略。     

Constitutive equations for dilute bubble suspensions and rheological behavior in simple shear and uniaxial elongational flow fields

A theoretical model is proposed in order to investigate rheological behavior of bubble suspension with large deformation. Theoretical constitutive equations for dilute bubble suspensions are derived by applying a deformation theory of ellipsoidal droplet [1] to a phenomenological suspension theory [2]. The rate of deformation tensor within the bubble and the time evolution of interface tensor are predicted by applying the proposed constitutive equations, which have two free fitting parameters. The transient and steady rheological properties of dilute bubble suspensions are studied for several capillary numbers (Ca) under simple shear flow and uniaxial elongational flow fields. The retraction force of the bubble caused by the interfacial tension increases as bubbles undergo deformation. The transient and steady relative viscosity decreases asCa increases. The normal stress difference (NSD) under the simple shear has the largest value whenCa is around 1 and the ratio of the first NSD to the second NSD has the value of 3/4 for largeCa but 2 for smallCa. In the uniaxial elongational flow, the elongational viscosity is three times as large as the shear viscosity like the Newtonian fluid.     

Prediction and measurement of residual stresses in injection molded parts

Residual stresses were predicted by a flow analysis in the mold cavity and residual stress distribution in the injection molded product was measured. Flow field was analyzed by the hybrid FEM/FDM method, using the Hele Shaw approximation. The Modified Cross model was used to determine the dependence of the viscosity on the temperature and the shear rate. The specific volume of the polymer melt which varies with the pressure and temperature fields was calculated by the Tait’s state equation. Flow analysis results such as pressure, temperature, and the location of the liquid-solid interface were used as the input of the stress analysis. In order to calculate more accurate gap-wise temperature field, a coordinate transformation technique was used. The residual stress distribution in the gap-wise direction was predicted in two cases, the free quenching and the constrained quenching, under the assumption that the shrinkage of the injection molded product occurs within the mold cavity and that the solid polymer is elastic. Effects of the initial flow rate, packing pressure, and mold temperature on the residual stress distribution was discussed. Experimental results were also obtained by the layer removal method for molded polypropylene.