Isolation，Identification and Characterization of Rhodobacter azotoformans SY5 with Enhancing Effect on the Disease Resistance of Eriocheir sinensis

doi:10.13560/j.cnki.biotech.bull.1985.2022-0043

Abstract

Abstract:

In order to enrich microbial resources with the potential application in the prevention and control of diseases in Eriocheir sinensis, strain SY5 of aerobic photosynthetic bacterium was isolated from the sediment of culture pond with carotenoid production ability as the evaluation index, was identified based on morphological, biochemical and molecular identification, and its antibiotic susceptibility and protective effect on E. sinensis against Citrobacter freundii infection were further evaluated. The results indicated that the SY5 isolate was identified phenotypically and molecularly as Rhodobacter azotoformans. It was Gram negative, formed colonies with the characteristics of dark red, round, smooth, moist and regular-edged on nutrient agar plate, showed the closest genetic relationship with R. azotoformans YLK20, and demonstrated highly sensitive to amoxicillin, neomycin, gentamicin, tetracycline, polymyxin B, enrofloxacin, ciprofloxacin, kanamycin, ampicillin, roxithromycin, fosfomycin, streptomycin and moderately sensitive to doxycycline. Besides, it demonstrated effective protection rates of 60.0% to 100% at final doses of 6.0×10⁵ to 6.0×10⁹ CFU/g diets against C. freundi infection in E. sinensis. These data lays a foundation for the later development of R. azotoformans SY5 as a feed additive for E. sinensis aquaculture.

Key words: Rhodobacter azotoformans, Eriocheir sinensis, characterization, Citrobacter freundii

CAO Hai-peng, ZHANG Shu-meng, DIAO Jing, XU La, GAI Chun-lei. Isolation，Identification and Characterization of Rhodobacter azotoformans SY5 with Enhancing Effect on the Disease Resistance of Eriocheir sinensis[J]. Biotechnology Bulletin, 2022, 38(11): 277-285.

Figures/Tables 6

References 35

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项目Test	结果Result		项目Test	结果Result
项目Test	SY5	固氮红细菌R. azotoformans^[13]	项目Test	SY5	固氮红细菌R. azotoformans^[13]
甲酸钠Sodium formate	+	+	半乳糖醇 Dulcitol	+	+
乙酸钠Sodium acetate	+	+	甘露醇Mannitol	+	+
丙酸钠Sodium propionate	-	-	柠檬酸钠Trisodium citrate dihydrate	+	+
甘油Glycerol	+	+	酒石酸钠Sodium tartaric	-	-
葡萄糖Glucose	+	+	葡萄糖酸钠Sodium citrate	+	+
果糖Fructose	+	+	乙醇Ethanol	+	+
过氧化氢酶Catalase	+	+	硫代硫酸钠Sodium thiosulfate	-	-
甘露糖Mannose	+	+	硫化钠Sodium sulfide	-	-
苯甲酸钠Sodium benzoic	-	-	3%NaCl生长 3%NaCl growth	+	+

项目Test	结果Result		项目Test	结果Result
项目Test	SY5	固氮红细菌R. azotoformans^[13]	项目Test	SY5	固氮红细菌R. azotoformans^[13]
甲酸钠Sodium formate	+	+	半乳糖醇 Dulcitol	+	+
乙酸钠Sodium acetate	+	+	甘露醇Mannitol	+	+
丙酸钠Sodium propionate	-	-	柠檬酸钠Trisodium citrate dihydrate	+	+
甘油Glycerol	+	+	酒石酸钠Sodium tartaric	-	-
葡萄糖Glucose	+	+	葡萄糖酸钠Sodium citrate	+	+
果糖Fructose	+	+	乙醇Ethanol	+	+
过氧化氢酶Catalase	+	+	硫代硫酸钠Sodium thiosulfate	-	-
甘露糖Mannose	+	+	硫化钠Sodium sulfide	-	-
苯甲酸钠Sodium benzoic	-	-	3%NaCl生长 3%NaCl growth	+	+

抗生素 Antibiotics	抑菌圈直径判断标准 Assessment standard of inhibition zone diameter /mm			药物含量 Content /（μg·disc^-1）	抑菌圈直径 Diameter of inhibition zone/mm	敏感性 Susceptibility
抗生素 Antibiotics	不敏感R Resistance	中度敏感I Intermediate sensitivity	高度敏感S Sensitivity	药物含量 Content /（μg·disc^-1）	抑菌圈直径 Diameter of inhibition zone/mm	敏感性 Susceptibility
阿莫西林Amoxicillin	≤13	14-17	≥18	10	24.30±0.78	S
新霉素Neomycin	≤12	13-16	≥17	30	24.05±0.83	S
庆大霉素Gentamicin	≤12	13-14	≥15	10	18.20±0.44	S
四环素Tetracycline	≤14	15-18	≥19	30	19.22±0.56	S
多黏菌素B Polymyxin B	≤8	9-11	≥12	30	15.21±0.82	S
恩诺沙星Enrofloxacin	≤15	16-18	≥19	5	26.55±0.70	S
环丙沙星Ciprofloxacin	≤15	16-20	≥21	5	24.20±0.65	S
卡那霉素Kanamycin	≤13	14-17	≥18	30	24.40±0.45	S
氨苄西林Ampicillin	≤13	14-16	≥17	10	22.50±0.25	S
罗红霉素Roxithromycin	≤10	11-15	≥16	15	16.50±1.06	S
磷霉素Fosfomycin	≤12	13-15	≥16	200	23.12±0.60	S
链霉素Streptomycin	≤11	12-14	≥15	10	25.25±0.85	S
多西环素Doxycycline	≤17	18-22	≥23	30	20.85±0.65	I

抗生素 Antibiotics	抑菌圈直径判断标准 Assessment standard of inhibition zone diameter /mm			药物含量 Content /（μg·disc^-1）	抑菌圈直径 Diameter of inhibition zone/mm	敏感性 Susceptibility
抗生素 Antibiotics	不敏感R Resistance	中度敏感I Intermediate sensitivity	高度敏感S Sensitivity	药物含量 Content /（μg·disc^-1）	抑菌圈直径 Diameter of inhibition zone/mm	敏感性 Susceptibility
阿莫西林Amoxicillin	≤13	14-17	≥18	10	24.30±0.78	S
新霉素Neomycin	≤12	13-16	≥17	30	24.05±0.83	S
庆大霉素Gentamicin	≤12	13-14	≥15	10	18.20±0.44	S
四环素Tetracycline	≤14	15-18	≥19	30	19.22±0.56	S
多黏菌素B Polymyxin B	≤8	9-11	≥12	30	15.21±0.82	S
恩诺沙星Enrofloxacin	≤15	16-18	≥19	5	26.55±0.70	S
环丙沙星Ciprofloxacin	≤15	16-20	≥21	5	24.20±0.65	S
卡那霉素Kanamycin	≤13	14-17	≥18	30	24.40±0.45	S
氨苄西林Ampicillin	≤13	14-16	≥17	10	22.50±0.25	S
罗红霉素Roxithromycin	≤10	11-15	≥16	15	16.50±1.06	S
磷霉素Fosfomycin	≤12	13-15	≥16	200	23.12±0.60	S
链霉素Streptomycin	≤11	12-14	≥15	10	25.25±0.85	S
多西环素Doxycycline	≤17	18-22	≥23	30	20.85±0.65	I

浓度 Concentration/（CFU · g^-1）	蟹数 Crab No. /ind.	累计死亡数Total number of deaths /ind.							平均死亡率 Average mortality/%
浓度 Concentration/（CFU · g^-1）	蟹数 Crab No. /ind.	1 d	2 d	3 d	4 d	5 d	6 d	7 d	平均死亡率 Average mortality/%
0	10	0	3	4	6	6	8	9	83.33±11.55
	10	0	2	3	3	5	7	9
	10	0	1	2	3	5	5	7
6.0×10⁵	10	0	1	1	1	2	2	2	33.33±11.55
	10	0	1	1	1	2	3	4
	10	0	0	1	1	1	2	4
6.0×10⁶	10	0	0	0	1	1	1	2	13.33±5.77
	10	0	0	0	0	0	1	1
	10	0	0	0	0	0	0	1
6.0×10⁷	10	0	0	0	0	0	0	1	6.66±5.77
	10	0	0	0	0	0	0	1
	10	0	0	0	0	0	0	0
6.0×10⁸	10	0	0	0	0	0	0	0	0±0
	10	0	0	0	0	0	0	0
	10	0	0	0	0	0	0	0
6.0×10⁹	10	0	0	0	0	0	0	0	0±0
	10	0	0	0	0	0	0	0
	10	0	0	0	0	0	0	0