中华鳖养殖水体的环境因子调查与机体致病菌研究
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摘要
对河北省阜平县中华鳖养殖基地的疾病情况及养殖水体的环境因子进行为期9个月的采样和监测分析,探讨了水体中浮游植物的群落特征、理化因子对中华鳖疾病的影响；从患病中华鳖体内分离纯化细菌并进行鉴定,得到了3株能够感染中华鳖的致病菌,找出了能够有效抑制致病菌的药物；进行回染实验,通过对感染的中华鳖血液进行生理生化分析,发现患病后许多指标出现异常变化。上述结果为有效预防中华鳖细菌性疾病的发生提供参考依据。
共鉴定出5门、5纲、22科、42种浮游植物,其中绿藻门藻类最多,有19属25种；其次为蓝藻门,9属13种；其它三门相对较少。优势种为四尾栅藻(Scenedesmus quadricanda),其较高的耐污性提示养殖水体有机污染程度较高。对养殖水体进行的化学指标监测结果表明总氮与氨氮含量比较高,结合疾病情况,可看出中华鳖的患病情况与总氮、氨氮含量的变化相一致。
分离出3株优势细菌,通过菌体和菌落形态观察、生理生化特征分析和16SrDNA测序,鉴定出该3种细菌分别为嗜水气单胞菌(Aeromonas hydrophila)、铜绿假单胞菌(Pseudomonas aeruginosa)、温和气单胞菌(Aeromonas sobria)。细菌回染实验结果表明3株细菌都能引起机体患病。药敏试验结果显示,嗜水气单胞菌对庆大霉素、红霉素、环丙沙星和氯霉素都有极强的敏感性；铜绿假单胞菌对四环素有极强的敏感性,对诺氟沙星、庆大霉素和环丙沙星有较强的敏感性；温和气单胞菌对土霉素、红霉素有极强的敏感性。在治疗中华鳖细菌性疾病时,可以选用针对性药物进行治疗,这样能在短时间内起到良好的效果。
中华鳖在感染致病菌后,血细胞数量大量减少；血液内谷丙转氨酶、谷草转氨酶、乳酸脱氢酶3种酶活力显著高于健康中华鳖,这可能是病原感染后造成中华鳖体内器官病变,刺激酶活力升高,同时细胞内酶大量逸出也能导致这种情况。肌酸激酶变化不显著,可见中华鳖患病后对肌肉细胞的影响不大。蛋白质代谢方面：病鳖血液中总蛋白、白蛋白含量下降,尿素氮含量显著提高,说明中华鳖免疫系统受到细菌感染影响,免疫功能严重下降,其中尿素氮含量增加的原因可能是中华鳖患病后肾单位滤过及重吸收功能失调,肾功能不全。脂类代谢方面：患病中华鳖血液中甘油三酯含量显著低于健康中华鳖,甘油三酯含量与肝脏的健康程度密切相关,可见患病中华鳖肝脏受到了严重损伤；总胆固醇含量变化不显著。离子代谢方面：Ca2+、Na+和Cl-含量变化不显著,这可能是由于患病时间较短,离子浓度还没有出现显著变化。
Nine months'sampling and monitoring analysis about diseases condition and aquaculture water of P. sinensis in Fuping P. sinensis farm were done. Effects of of water quality and community character of phytoplankton in the aquaculture water were discussed; Three strains of bacteria were isolated from the diseased P. sinensis. According to the artifically infection experiment, they were the pathogenic bacteria of P. sinensis. A few of medicines were selected through the drug sensitivity test which can inhibit the pathogenic bacteria effectively; Physiology and biochemistry analysis on the infected blood of P. sinensis showed that there were abnormal changes in blood indexes of diseased P. sinensis, which is helpful to the prevention of P. sinensis's diseases.
42 species of phytoplankton were identified, which belongs to 5 phylum,5 class,22 family. Among them, Chlorophyta includes the most,25 species, which belong to 19 genera; Cyanophyta ranked secondly,13 species, which belong to 9 genera. The dominant species of phytoplankton in aquaculture water of P. sinensis was S. quadricanda. Due to the pollution resistance character of S. quadricanda, It could be drawn that the aquaculture water of P. sinensis organic pollution degree was comparatively high. The result of chemical indexes monitoring about the aquaculture water showed that the content of total nitrogen and ammonia nitrogen was comparatively high. Associating that with the diseases condition, we could draw the conclusion that there was positive correlation between the diseases condition and the change of content of total nitrogen and ammonia nitrogen.
In the isolation and identification of bacteria, three dominant strains were isolated. They are A. hydrophila, P. aeruginosa, and A. sobria, respectively. According to the artificially infection experiment, they were the pathogenic bacteria of P. sinensis. The drug sensitivity test showed that A. hydrophila was extremely sensitive to gentamicin, erythromycin, ciprofloxacin, and chloramphenicol. Pseudomonas was extremely sensitive to tetracycline, and it's sensitivity to norfloxacin, gentamicin, and ciprofloxacin was comparatively high. A. sobria was extremely sensitive to oxytetracycline and erythromycin. Treating bacteria-infected P. sinensis with the pertinent medicine could have well therapeutic effect in short time.
The blood cells of P. sinensis infected by pathogenic bacteria decreased dramatically in number. With reference to enzyme activity:the enzyme activity of AST, ALT, and LDH of diseased P. sinensis were remarkably higher than those of the healthy one.The diseased P. sinensis was stimulate by pathogenic bacteria, which lead to organ lesions, could explain the change of enzyme activity. In the meantime, the endoenzyme had emit could also be the possible reasons for the change. The change of CK enzyme activity was not significant. About protein metabolism:the contents of total protein and albumin decreased, and the contents of urea nitrogen increased significantly. It showed that the immune system of P. sinensis was affected by bacteria, and thus the immune function decreased tremendously. The filtration and reabsorption dysfunction of nephron of diseased P. sinensis resulted in the increase of urea nitrogen contents. As to lipid metabolism:the content of triglyceride of diseased P. sinensis was significantly lower than that of the healthy one. Because the content of triglyceridethe is closely related to the normal function of liver, the liver of diseased P. sinensis was severely damaged. The content of CHOL changed a little. Concerning ion metabolism:the change of contents of Ca2+, Na+, Cl- were not significant. Relatively short sick time might explain this.
共鉴定出5门、5纲、22科、42种浮游植物,其中绿藻门藻类最多,有19属25种；其次为蓝藻门,9属13种；其它三门相对较少。优势种为四尾栅藻(Scenedesmus quadricanda),其较高的耐污性提示养殖水体有机污染程度较高。对养殖水体进行的化学指标监测结果表明总氮与氨氮含量比较高,结合疾病情况,可看出中华鳖的患病情况与总氮、氨氮含量的变化相一致。
分离出3株优势细菌,通过菌体和菌落形态观察、生理生化特征分析和16SrDNA测序,鉴定出该3种细菌分别为嗜水气单胞菌(Aeromonas hydrophila)、铜绿假单胞菌(Pseudomonas aeruginosa)、温和气单胞菌(Aeromonas sobria)。细菌回染实验结果表明3株细菌都能引起机体患病。药敏试验结果显示,嗜水气单胞菌对庆大霉素、红霉素、环丙沙星和氯霉素都有极强的敏感性；铜绿假单胞菌对四环素有极强的敏感性,对诺氟沙星、庆大霉素和环丙沙星有较强的敏感性；温和气单胞菌对土霉素、红霉素有极强的敏感性。在治疗中华鳖细菌性疾病时,可以选用针对性药物进行治疗,这样能在短时间内起到良好的效果。
中华鳖在感染致病菌后,血细胞数量大量减少；血液内谷丙转氨酶、谷草转氨酶、乳酸脱氢酶3种酶活力显著高于健康中华鳖,这可能是病原感染后造成中华鳖体内器官病变,刺激酶活力升高,同时细胞内酶大量逸出也能导致这种情况。肌酸激酶变化不显著,可见中华鳖患病后对肌肉细胞的影响不大。蛋白质代谢方面：病鳖血液中总蛋白、白蛋白含量下降,尿素氮含量显著提高,说明中华鳖免疫系统受到细菌感染影响,免疫功能严重下降,其中尿素氮含量增加的原因可能是中华鳖患病后肾单位滤过及重吸收功能失调,肾功能不全。脂类代谢方面：患病中华鳖血液中甘油三酯含量显著低于健康中华鳖,甘油三酯含量与肝脏的健康程度密切相关,可见患病中华鳖肝脏受到了严重损伤；总胆固醇含量变化不显著。离子代谢方面：Ca2+、Na+和Cl-含量变化不显著,这可能是由于患病时间较短,离子浓度还没有出现显著变化。
Nine months'sampling and monitoring analysis about diseases condition and aquaculture water of P. sinensis in Fuping P. sinensis farm were done. Effects of of water quality and community character of phytoplankton in the aquaculture water were discussed; Three strains of bacteria were isolated from the diseased P. sinensis. According to the artifically infection experiment, they were the pathogenic bacteria of P. sinensis. A few of medicines were selected through the drug sensitivity test which can inhibit the pathogenic bacteria effectively; Physiology and biochemistry analysis on the infected blood of P. sinensis showed that there were abnormal changes in blood indexes of diseased P. sinensis, which is helpful to the prevention of P. sinensis's diseases.
42 species of phytoplankton were identified, which belongs to 5 phylum,5 class,22 family. Among them, Chlorophyta includes the most,25 species, which belong to 19 genera; Cyanophyta ranked secondly,13 species, which belong to 9 genera. The dominant species of phytoplankton in aquaculture water of P. sinensis was S. quadricanda. Due to the pollution resistance character of S. quadricanda, It could be drawn that the aquaculture water of P. sinensis organic pollution degree was comparatively high. The result of chemical indexes monitoring about the aquaculture water showed that the content of total nitrogen and ammonia nitrogen was comparatively high. Associating that with the diseases condition, we could draw the conclusion that there was positive correlation between the diseases condition and the change of content of total nitrogen and ammonia nitrogen.
In the isolation and identification of bacteria, three dominant strains were isolated. They are A. hydrophila, P. aeruginosa, and A. sobria, respectively. According to the artificially infection experiment, they were the pathogenic bacteria of P. sinensis. The drug sensitivity test showed that A. hydrophila was extremely sensitive to gentamicin, erythromycin, ciprofloxacin, and chloramphenicol. Pseudomonas was extremely sensitive to tetracycline, and it's sensitivity to norfloxacin, gentamicin, and ciprofloxacin was comparatively high. A. sobria was extremely sensitive to oxytetracycline and erythromycin. Treating bacteria-infected P. sinensis with the pertinent medicine could have well therapeutic effect in short time.
The blood cells of P. sinensis infected by pathogenic bacteria decreased dramatically in number. With reference to enzyme activity:the enzyme activity of AST, ALT, and LDH of diseased P. sinensis were remarkably higher than those of the healthy one.The diseased P. sinensis was stimulate by pathogenic bacteria, which lead to organ lesions, could explain the change of enzyme activity. In the meantime, the endoenzyme had emit could also be the possible reasons for the change. The change of CK enzyme activity was not significant. About protein metabolism:the contents of total protein and albumin decreased, and the contents of urea nitrogen increased significantly. It showed that the immune system of P. sinensis was affected by bacteria, and thus the immune function decreased tremendously. The filtration and reabsorption dysfunction of nephron of diseased P. sinensis resulted in the increase of urea nitrogen contents. As to lipid metabolism:the content of triglyceride of diseased P. sinensis was significantly lower than that of the healthy one. Because the content of triglyceridethe is closely related to the normal function of liver, the liver of diseased P. sinensis was severely damaged. The content of CHOL changed a little. Concerning ion metabolism:the change of contents of Ca2+, Na+, Cl- were not significant. Relatively short sick time might explain this.
引文
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