蜂胶在食品保鲜和畜禽上的应用前景

3畜禽疾病的防治 蜂胶是特异性和非特异性免疫因子的激活剂，能促进抗体的产生，激活生物体自身的免疫防卫系统，增强巨噬细胞的吞噬能力，提高肌体对有害物质侵入的抵抗力。我国兽医研究人员用蜂胶作为佐剂研制的畜禽疫苗，可增强疫苗的免疫力。     

蜂胶,蜂花粉复合微量元素添加剂喂猪试验

现代畜牧营养科学正向着无毒、无公害、无“三致因子”、无残留、高效完善方面发展,以能生产出高质量的畜产品,并提高经济效益。研究表明:蜂产品——蜂胶、蜂花粉符合这种需求,成为营养学者关注的课题。 国外大量研究表明:蜂胶具有较强的辅佐抗原免疫作用,能增强抗体的产生,使血清总数CP和r-球蛋白含量增加,增强白细胞和巨噬细胞的吞噬能力,从而提高机体的免疫力。可以认为:蜂胶对感染性疾病的疗效一方面是由于其具有抗菌、抗原虫、抗病毒作用,抑制了致病微生物的生长和繁殖;另一方面则由于其增强了机体的特异性和非特异性免疫能力,提高机体的     

瑞博·康思农蜂胶片对小鼠细胞、体液、巨噬细胞免疫力影响

以提纯蜂胶为主料、蜂花粉为辅料加工而成的蜂胶片分成低剂量、中剂量、高剂量,对受试动物昆明种小鼠进行试验,小鼠被随机分别分为阴性对照组及低、中、高3个剂量组,每天1次按0.4mg/20g体重,连续30d.研究了蜂胶片对小鼠细胞、体液免疫力的提高效果与巨噬细胞吞噬能力的提高效果,试验结果表明,瑞博·康思农蜂胶片具有明显增强细胞和体液免疫功能的作用,增强巨噬细胞吞噬作用.     

蜂胶对淋巴细胞调节作用的研究进展

综述了蜂胶对特异性免疫系统影响的国内外研究进展,特别是蜂胶对抗体和淋巴因子分泌、淋巴细胞增殖和分化的调控.结果表明:蜂胶可促进抗体的产生,促进免疫低下的机体和接种疫苗或肿瘤细胞的机体淋巴细胞的增殖,增强机体免疫力;蜂胶能调节淋巴细胞分化和淋巴因子的产生,抑制淋巴细胞的增殖,与其抗炎特性相关.     

抗菌肽饲料添加剂对小鼠巨噬细胞吞噬功能的影响

对试验小鼠饲喂1‰和3‰抗菌肽饲料添加剂,鸡红细胞吞噬试验检测小鼠腹腔巨噬细胞吞噬功能,研究抗菌肽饲料添加剂对小鼠免疫功能的影响.结果表明,抗菌肽饲料添加剂可以显著提高小鼠腹腔巨噬细胞吞噬鸡红细胞的吞噬率和吞噬指数,增强小鼠非特异性免疫功能.     

鸡外周血单核/巨噬细胞吞噬能力的研究

单核/巨噬细胞在沟通非特异性免疫与特异性免疫和提高机体免疫力、抗病力方面具有重要的意义.试验对农大褐3号节粮型蛋鸡29周龄产蛋高峰期单核细胞吞噬特性进行研究,揭示单核/巨噬细胞分离、纯化的一般方法和其吞噬现象的内在本质规律,以期为机体提高抗病能力、开展抗病育种提供新思路、新方法.     

羊胎盘转移因子注射液非特异性免疫调节作用的研究

为探讨羊胎盘转移因子注射液(goat placental transfer factor injection,GPTF)对动物非特异性免疫功能的影响,采用巨噬细胞吞噬鸡红细胞试验、碳廓清试验以及诱导靶细胞释放免疫介质试验,评价羊胎盘转移因子注射液对动物非特异性免疫调节的作用。结果显示,羊胎盘转移因子注射液具有促进巨噬细胞吞噬鸡红细胞功能、提高受试小鼠免疫系统廓清指数和吞噬指数以及增加巨噬细胞一氧化氮(NO)和白细胞介素-1(IL-1)分泌量的作用。试验结果表明,羊胎盘转移因子注射液具有增强动物机体非特异性免疫力的作用。     

绿曼"蜂胶百毒净"——打造动保行业新理念

<正>为了有效控制疾病,保护动物健康,青岛绿曼公司经过三年艰苦的摸索,通过"复方蜂胶溶液"在临床上对畜禽的保健推广及成功的治疗经验,终于第一个成功的总结出:用蜂胶重构动物非特异性主动免疫功能     

蜂胶醇提取物增强小鼠免疫功能研究

试验表明、蜂胶醇提取物能显著提高小鼠碳粒廓清速度（Ｋ），能明显促进鸡红细胞致敏小鼠溶血素的生成和提高胸腺系数，具有提高健康小鼠特异性和非特异性免疫功能的作用。     

禽霍乱+大肠杆菌病二联蜂胶灭活疫苗与油佐剂疫苗注射部位的动态病理组织学比较观察

用禽霍乱+大肠杆菌二联蜂胶灭活疫苗和油佐剂疫苗分别胸肌注射免疫肉鸡,分别于免疫后7d、14d和28d剖杀,取注射部位的肌肉组织,常规石蜡包埋切片观察。结果为,蜂胶疫苗免疫组免疫后7d时,肌纤维轻度水肿,纤维间有大量淋巴细胞和巨噬细胞的浸润,巨噬细胞吞噬有蜂胶颗粒,14d时肌纤维则恢复正常;油苗注射组免疫后,7d时肌肉中度水肿,其间有大量油乳状疫苗残留,镜检可见肌纤维断裂、坏死,其间有较多较大的油囊,囊壁由大量增生的成纤维细胞、异嗜性粒细胞、巨噬细胞、小血管及淋巴样细胞组成,细胞增生持续存在到28d,最后形成肉芽肿结构。研究结果表明,蜂胶疫苗免疫后对局部组织学影响明显小于油苗注射组。     

Macrophages in central nervous canine distemper: friends or foes?

Brain macrophages play an important role in the CNS. We review some biological aspects of brain macrophages in vivo and in vitro. The criteria and methods used for the identification of these cells are considered. They include some morphological features such as the use of histochemical and immunocytochemical techniques for internal and surface components. In the second part of this review, we describe the role of brain macrophages in canine distemper encephalitis as proposed earlier by us. Studies in cultured dog glial cells infected with canine distemper virus (CDV) have shown that brain macrophages stimulated by anti-CDV antibodies will release reactive oxygen species as measured by chemiluminescence. This response depends on the presence of viral antigens on the surfaces of infected cells and is mediated by the interaction of antigen-bound antibodies with Fc receptors on brain macrophages. These observations support the hypotheses that brain macrophages may contribute to the damage of the white matter observed in canine distemper encephalitis.     

Macrophages from chickens selected for high antibody response produced more nitric oxide and have greater phagocytic capacity

Macrophages are fundamental cells of the innate immune system, which, through phagocytosis and nitric oxide production, eliminate pathogens. The aim of the present study was to determine if macrophages from chicken families divergently selected to high and low antibodies response differ in nitric oxide production and phagocytic capacity. Blood monocytes derived macrophages were activated with lipopolysaccharide and supernatant from chicken spleen lymphocytes cultured with Concanavalin A (containing chicken interferon). Nitric oxide production was evaluated in culture supernatants. Phagocytic capacity of activated and non-activated macrophages was assayed using yeasts and IgY opsonized sheep red blood cells. Activated and non-activated macrophages from the high antibodies response family produced higher nitric oxide levels, internalized more yeast and significantly more opsonized sheep red blood cells than macrophages from the low antibodies response family. Moreover, activated macrophages became more elongated and widely spread. These findings indicate that macrophages from the high antibodies response family were more active suggesting that the differences in antibody response also depend on macrophage function.     

Pathological, ultrastructural, and immunohistochemical changes caused by Lelystad virus in experimentally induced infections of mystery swine disease (synonym: porcine epidemic abortion and respiratory syndrome (PEARS))

The pathogenicity and pathogenesis of Lelystad virus was studied in six 6-day-old SPF piglets. A third passage of the agent was propagated on porcine alveolar macrophages and intranasally inoculated into pigs. Pigs were killed at hours 24, 48, 60, and 72, and on days 6 and 8 after inoculation. From day 2 on pigs developed diffuse interstitial pneumonia with focal areas of catarrhal pneumonia, and from this day on splenic red pulp macrophages were enlarged and vacuolated. Lelystad virus was re-isolated from the lungs of infected pigs from day 2 after inoculation. Lelystad virus antigens were detected by immunohistochemical techniques in bronchiolar epithelium and alveolar cells, and in spleen cells of infected pigs from day 2 after inoculation. Ultrastructural examination of tissues by electron microscopy revealed degenerating alveolar macrophages and epithelial cells in lungs and nasal mucosa, with excessive vacuolation of the endoplasmic reticulum. Although the respiratory tract seems to be the target organ for this virus, macrophages in other organs, such as the spleen, can also be infected. This preference for macrophages may impair immunological defences.     

Cell-mediated immune protection in chickens against Pasteurella multocida

Immune protection by cellular immunity in chickens against Pasteurella multocida was investigated by in vivo and in vitro experiments using spleen cells and culture supernatants of immunised chickens. Intraperitoneal or intravenous transfer of immune splenic cells into normal chickens induced transmission of an as effective protection as that exhibited in immunised chickens. Immune protection was also obtained by intravenous treatment of chickens with culture supernatant fluid from immune splenic cells of hormonally bursectomised chickens. The in vitro experiment showed that intracellular bacterial proliferation was inhibited in peritoneal macrophages from immunised chickens, or from normal chickens sensitised with culture supernatant fluid of immune splenic cells, and the macrophages were protected from disruption by infection. Peritoneal macrophages sensitised with culture supernatant fluid from unimmunised splenic cells, or peritoneal macrophages from unimmunised chickens, allowed considerable intracellular proliferation of bacteria with almost complete breakdown of the macrophages within 24 hours after bacterial challenge. These data suggest that the protective immunity of chickens against P multocida was dependent on cell-mediated immunity by mediators such as the macrophage activating factor from T lymphocytes.     

Isolation and characterization of macrophages from a mixed primary culture of bovine liver cells

Previously, we developed a simple and efficient method to isolate liver macrophages from a mixed primary culture of adult rat liver cells. To extend the applicability of this method, we isolated macrophages from mixed primary cultures of bovine liver cells. Macrophage cells proliferated on the cell sheet of mixed bovine liver cells after 8-16d of culture. These cells were detached by shaking of the culture flasks. Subsequent transfer and brief incubation in plastic dishes resulted in selective adhesion of macrophages. After rinses with PBS, attached macrophages were harvested. More than 10(6) cells could be harvested from the culture flask at intervals of 2-3d for more than three weeks. The isolated cells were strongly positive for bovine macrophage markers, such as CD68, CD172a and Iba-1. These cells exhibited functional properties of macrophages, including active phagocytosis of polystyrene microbeads, proliferative response to recombinant bovine granulocyte-macrophage colony-stimulating factor, upregulation of specific inflammatory cytokine genes upon stimulation with lipopolysaccharide, and formation of multinucleated giant cells. The shaking and attachment method provides a simple and efficient alternative to obtain bovine liver macrophages without requiring complex equipment or specialized technical skills.     

Failure of antigen-stimulated gammadelta T cells and CD4+ T cells from sensitized cattle to upregulate nitric oxide and mycobactericidal activity of autologous Mycobacterium avium subsp. paratuberculosis-infected macrophages

The function of gammadelta T cells during ruminant paratuberculosis (Johne's disease) is presently unknown. An ex vivo system was used to test the hypothesis that gammadelta T cells are capable of activating Mycobacterium avium subsp. paratuberculosis-(M. paratuberculosis)-infected macrophages. Peripheral blood-derived macrophages were infected in vitro with live M. paratuberculosis, and autologous LN-derived gammadelta T cells or CD4+ T cells were co-cultured with infected macrophages for 48h, at which time bacterial survival as well as production of nitrites and IFN-gamma was evaluated. Incubation of M. paratuberculosis-infected macrophages with autologous gammadelta T cells did not result in reduced intracellular bacterial viability compared to infected macrophage cultures without added T cells. IFN-gamma production by-infected cultures containing added gammadelta T cells was not enhanced compared to that of infected macrophages alone. Although infection of macrophage cultures caused increased production of nitrites at both post-infection day (PID) 0 and PID 60, the addition of gammadelta T cells did not further increase nitrite production. In contrast, addition of PPD-stimulated CD4+ T cells obtained at PID 60 to M. paratuberculosis-infected macrophages resulted in significantly increased IFN-gamma production compared to cultures without added T cells or cultures containing unstimulated CD4+ T cells or unstimulated or antigen-stimulated gammadelta T cells. However, the increased production of IFN-gamma by co-cultures containing PPD-stimulated CD4+ T cells did not result in increased bacterial killing or increased production of nitrites compared to cultures without added T cells. In additional in vitro experiments, M. paratuberculosis-infected macrophages, but not uninfected macrophages, were unable to increase nitrite production when stimulated with recombinant IFN-gamma. Taken together, the data suggest that (1) gammadelta T cells do not produce significant IFN-gamma and do not significantly increase NO production from M. paratuberculosis-infected macrophages in vitro, (2) the production of significant IFN-gamma by antigen-stimulated CD4+ T cells from infected calves is insufficient to enhance mycobacterial killing or nitrite production by infected macrophages, and (3) macrophages may have an impaired NO response following intracellular M. paratuberculosis infection, even in the presence of significant concentrations of IFN-gamma.     

Monoclonal antibodies against porcine macrophages

Two mouse monoclonal antibodies (MAB) (clones 2G6 and 2B10) directed against porcine macrophages are described that are suitable for use in immunohistochemistry, FACS analysis and western blot. As immunogen, porcine cells from bronchoalveolar lavage (BAL) were used. The MABs obtained belonged to the mouse IgG1 subclass. The molecular weights of the corresponding antigens were detected by western blot under non-reducing conditions (2G6: 140-150kDa; 2B10: 140-145kDa). For specificity screening, porcine snap-frozen tissues of lung, lung lymph node, tonsil, spleen, thymus, brain, liver, gut and kidney were used. The MABs were able to identify cell populations of the mononuclear phagocytic system in these organs. While MAB 2G6 detected tissue macrophages (sinusoidal lymph node macrophages, red pulp spleen macrophages, Kupffer cells, Langerhans cells, thymus macrophages, macrophages of lung and macrophages of kidney), MAB 2B10 stained cells scattered in the lymph node (subsinusoidal, interfollicular and follicular macrophages) and in the lung interstitium. Additionally, it showed reactivity with Kupffer cells, spleen and kidney macrophages. An immunoreactivity of the MABs could be established also for human but not for bovine and avian macrophages. By flow cytometric analysis, MAB 2B10 reacted with a subpopulation of BAL and peritoneal cells. Antibody 2G6 detected macrophages of the BAL and the peritoneal fluid as well as peripheral blood monocytes.     

Iron modulates the replication of virulent Mycobacterium bovis in resting and activated bovine and possum macrophages

Bovine and possum macrophages were infected in vitro with a virulent strain of Mycobacterium bovis, and mycobacterial replication was measured in the infected macrophages cultured under a variety of conditions. Virulent M. bovis replicated substantially in alveolar possum macrophages as well as in bovine blood monocyte-derived macrophages. Addition of recombinant bovine interferon-gamma (IFN-gamma) with low concentrations of lipopolysaccharide (LPS) rendered bovine macrophages significantly more resistant to M. bovis replication. Disruption of iron levels in infected macrophages by addition of apotransferrin or bovine lactoferrin blocked replication of M. bovis in both bovine and possum macrophages. On the other hand, addition of exogenous iron, either in the form of iron citrate or iron-saturated transferrin, rendered macrophages of both species much more permissive for the replication of M. bovis. The impact of iron deprivation/loading on the mycobacteriostatic activity of cells was independent of nitric-oxide release, as well as independent of the generation of oxygen radical species in both possum and bovine macrophages. Exogenous iron was shown to reverse the ability of IFN-gamma/LPS pulsed bovine macrophages to restrict M. bovis replication. When autologous possum lymphocytes from animals vaccinated with M. bovis strain BCG were added to infected macrophages, they rendered the macrophages less permissive for virulent M. bovis replication. Loading the cells with iron prior to this macrophage-lymphocyte interaction, reversed this immune effect induced by sensitized cells. We conclude that, in two important animal species, intracellular iron level plays an important role in M. bovis replication in macrophages, irrespective of their activation status.     

Activation of macrophages by culture fluid of antigen-stimulated spleen cells collected from chickens immunized with Eimeria tenella

The effect of spleen cell factors on the activation of macrophages was investigated in chickens immunized with Eimeria tenella. The abilities of peritoneal macrophages obtained from normal chickens to kill sporozoites of E. tenella and to inhibit intracellular development of Toxoplasma gondii were enhanced by exposing them to 33 and 50% culture fluid of antigen-stimulated spleen cells of chickens immunized with E. tenella. The parasiticidal activity of normal macrophages was also distinctly enhanced by the treatment with culture fluid of phytohemagglutinin-stimulated normal spleen cells. On the other hand, the parasiticidal activity of normal macrophages could not be enhanced by the treatment with culture fluid of antigen-stimulated normal spleen cells under conditions similar to those of culture fluid of antigen-stimulated immune spleen cells. It thus appears that the macrophage activator was induced from immune spleen cells in response to the stimulation by the antigen.