本试验旨在通过研究异常毕赤酵母AR₂₀₁₆菌株对昆明小鼠生长性能、血清生化指标、血常规和免疫功能的影响，综合评价异常毕赤酵母AR₂₀₁₆菌株的安全性。试验1：选择20只健康的昆明小鼠，随机分为2组，每组10只，雌雄各占1/2。按每10 g体重最大灌胃容量0.2 mL进行7 d经口毒性试验，对照组灌胃生理盐水，试验组灌胃异常毕赤酵母AR₂₀₁₆菌株发酵液（菌体浓度2×10⁹ CFU/mL）。试验2：选择40只健康的昆明小鼠，随机分为4组，每组10只，雌雄各占1/2。每10 g体重最大灌胃容量0.2 mL进行28 d经口毒性试验，对照组灌胃生理盐水，低、中、高剂量组灌胃异常毕赤酵母AR₂₀₁₆菌株发酵液，菌体浓度分别为2×10⁷、2×10⁸、2×10⁹ CFU/mL。结果表明：1）异常毕赤酵母AR₂₀₁₆菌株处理7或28 d后，小鼠无明显中毒现象，未出现死亡，属"无毒"级（最大耐受剂量>20 000 mg/kg）。2）与对照组相比，异常毕赤酵母AR₂₀₁₆菌株处理7 d后，试验组小鼠平均日采食量（ADFI）显著降低（P<0.05）；异常毕赤酵母AR₂₀₁₆菌株处理28 d后，各剂量组小鼠ADFI显著提高（P<0.05）。3）与对照组相比，异常毕赤酵母AR₂₀₁₆菌株处理7 d后，试验组小鼠血清白蛋白、尿素氮含量显著降低（P<0.05），血清肌酐含量和谷草转氨酶活性显著提高（P<0.05）；异常毕赤酵母AR₂₀₁₆菌株处理28 d后，各组间血清生化指标无显著变化（P>0.05）。4）异常毕赤酵母AR₂₀₁₆菌株处理7 d后，与对照组相比，试验组小鼠白细胞、红细胞压积及淋巴细胞数量显著提高（P<0.05），试验组雄性小鼠血红蛋白含量显著高于对照组雄性小鼠（P<0.05），试验组雄性小鼠红细胞数量显著高于试验组雌性小鼠（P<0.05），试验组雌性小鼠中性细胞比率显著低于对照组雌性小鼠（P<0.05），试验组雌性小鼠嗜酸性粒细胞数量显著高于对照组雌性小鼠（P<0.05）；异常毕赤酵母AR₂₀₁₆菌株处理28 d后，各组间血常规指标无显著变化（P>0.05）。5）与对照组相比，异常毕赤酵母AR₂₀₁₆菌株处理7 d后，试验组小鼠血清中干扰素-γ（INF-γ）、肿瘤坏死因子-α含量显著提高（P<0.05）；异常毕赤酵母AR₂₀₁₆菌株处理28 d后，高剂量组雌性小鼠血清INF-γ含量显著提高（P<0.05）。综上，异常毕赤酵母AR₂₀₁₆菌株对昆明小鼠无毒，具较高生物安全性，可提高昆明小鼠ADFI，增强机体免疫功能，具有一定的开发利用价值。

To comprehensively evaluate the bio-safety of Pichia anonmala AR₂₀₁₆ strain, this study was carried out to investigate effects of Pichia anonmala AR₂₀₁₆ strain on growth performance, serum biochemical indexes, hemogram indexes and immune function in Kunming mice. In experiment 1, a total of 20 Kunming mice were randomly assigned to one of two groups with 10 mice in each group (half male and half female). A 7-day oral toxicity test was performed at a dose of 0.2 mL per 10 g body weight; control group supplemented with normal saline, and experimental group supplemented with Pichia anonmala AR₂₀₁₆ strain at the concentration of 2×10⁹ CFU/mL. In experiment 2, a total of 40 Kunming mice were randomly assigned to one of four groups with 10 mice in each group (half male and half female). A 28-day oral toxicity test was performed at a dose of 0.2 mL per 10 g body weight; control group supplemented with normal saline, low dose group, middle dose group and high dose group supplemented with Pichia anonmala AR₂₀₁₆ strain at concentrations of 2×10⁷, 2×10⁸, 2×10⁹ CFU/mL, respectively. The results showed as follows:1) after 7 or 28 d treatment by Pichia anonmala AR₂₀₁₆ strain, no poisoning and death were found in mice, it was a "non-toxic" grade, and the maximum tolerated dose was more than 20 000 mg/kg. 2) Compared with the control group, after 7 d treatment by Pichia anonmala AR₂₀₁₆ strain, average daily feed intake (ADFI) of mice in experimental group significantly decreased (P<0.05); after 28 d treatment by Pichia anonmala AR₂₀₁₆ strain, ADFI of mice in each dosage group significantly increased (P<0.05). 3) Compared with the control group, after 7 d treatment by Pichia anonmala AR₂₀₁₆ strain, the contents of albumin and urea nitrogen in serum of mice in experimental group significantly decreased (P<0.05), and serum creatinine content and aspartate transaminase activity significantly increased (P<0.05); after 28 d treatment by Pichia anonmala AR₂₀₁₆ strain, serum biochemical indexes among groups had no significant difference (P>0.05). 4) After 7 d treatment by Pichia anonmala AR₂₀₁₆ strain, compared with the control group, white blood cell, hematokrit, lymphocytes count of mice in experimental group significantly increased (P<0.05), hemoglobin content of male mice in experimental group was significantly higher than that of male mice in control group (P<0.05), red blood cell count of male mice in experimental group was significantly higher than that of female mice in experimental group (P<0.05), neutral cell rate of female mice in experimental group was significantly lower than that of female mice in control group (P<0.05), and count of eosinophils of female mice in experimental group was significantly higher than that of female mice in control group (P<0.05); after 28 d treatment by Pichia anonmala AR₂₀₁₆ strain, blood routine indexes among groups had no significant difference (P>0.05). 5) Compared with the control group, after 7 d treatment by Pichia anonmala AR₂₀₁₆ strain, the content of interferon-γ (INF-γ) and tumor necrosis factor-α in serum of mice in experimental group significantly increased (P<0.05); after 28 d treatment by Pichia anonmala AR₂₀₁₆ strain, INF-γ content in serum of female mice in high dose group significantly increased (P<0.05). Therefore, the Pichia anomala AR₂₀₁₆ strain is non-toxic and high safety to Kunming mice, can increase ADFI, improve immune function, and having the value for further research and development.