Oxoid CM0833B 气单胞菌培养基基础AEROMONAS MED BASE (RYAN) SUPP

Oxoid CM0833B 气单胞菌培养基基础AEROMONAS MED BASE (RYAN) SUPP

Dehydrated Culture Media

AEROMONAS MEDIUM BASE (RYAN)

Code: CM0833

A selective diagnostic medium for the isolation of Aeromonas hydrophila from clinical and environmental specimens when used with Ampicillin Selective Supplement.

当与氨苄青霉素选择性补充剂一起使用时,用于从临床和环境标本中分离嗜水气单胞菌的选择性诊断介质。

Typical Formula*

gm/litre

Proteose peptone

5.0

Yeast extract

3.0

L. Lysine monohydrochloride

3.5

L. Arginine monohydrochloride

2.0

Sorbitol

3.0

Inositol

2.5

Lactose

1.5

Xylose

3.75

Bile Salts No.3

3.0

Sodium thiosulphate

10.67

Sodium chloride

5.0

Ferric ammonium citrate

0.8

Bromothymol blue

0.04

Thymol blue

0.04

Agar

12.5

Final pH 8.0 + 0.2 @ 25°C

* Adjusted as required to meet performance standards

AMPICILLIN SELECTIVE SUPPLEMENT

Code: SR0136

Vial contents (each vial is sufficient for 500 ml of medium)
per vial
per litre
Ampicillin
2.5mg

5.0mg

Directions
Suspend 29.5g in 500ml of distilled water. Bring gently to the boil. DO NOT AUTOCLAVE. Cool to 50°C and aseptically add one vial of Ampicillin Selective Supplement SR0136 reconsituted as directed. Mix well and pour plates.

Description
Ryan1modified the formulation of XLD Medium so that it would support the growth of Aeromonas spp and Plesiomonas spp as well as the usual Enterobacteriaceae. It could therefore be used as a universal medium in the investigation of enteric disease. However, to improve its performance in the isolation of aeromonads, the addition of ampicillin at 5 mg/l is recommended. The effectiveness of ampicillin as a selective agent for Aeromonas spp has been reported by several workers 2,3,4,5,6. The value of Aeromonas Medium Base (Ryan) is that the recommended level of ampicillin is well below that which can cause inhibition of some strains of aeromonad7.

The utility of Aeromonas Medium (Ryan) and its superiority over some other formulae for detection of Aeromonas spp. in tap water, bottled water and foods including meat, poultry, fish and seafoods has been reported8,9,10. Aeromonas Medium (Ryan) is specified by the MAFF/DHS Steering Group on the Microbiological Safety of Food for detection and enumeration of Aeromonas hydrophila in clinical specimens11. Aeromonas spp occur widely in soil and water, where they cause diseases in fish and amphibians. They also occur in untreated and chlorinated drinking water, raw foods and raw milk11,12 . It is considered that the major cause of gastrointestinal infections by Aeromonas spp12,13 is from ingesting infected water14,15.

The role of these organisms in gastrointestinal disease is still subject to debate but a rapidly expanding body of literature suggests that Aeromonas spp can cause a wide spectrum of enteric symptoms in adults as well as children5,16 It would therefore be a useful diagnostic aid to include this selective medium when investigating diarrhoeal disease.

Aeromonas Medium Base has been developed to improve the enumeration and isolation of Aeromonas spp from clinical and environmental specimens.

Technique
1. Prepare the medium according to directions and pour into sterile dishes. The prepared medium may be stored at 2-8°C up to 5 days.
2. Inoculate the plates with a suspension of food, faeces etc., diluted to form single colonies on the inoculated plate.
3. Incubate the plates aerobically at 30-35°C for 24 hours. If further incubation is required hold at room temperature (22-25°C).
4. Examine the plates for the presence of dark green, opaque colonies with darker centres. Confirm the identity with biochemical tests.

方向
将 29.5g 悬浮在 500ml 蒸馏水中。轻轻煮沸。不要高压灭菌。冷却至 50°C 并在无菌条件下加入一瓶按照指示重新配制的氨苄青霉素选择性补充剂 SR0136。充分混合并倒入盘子。

描述
Ryan1 修改了 XLD 培养基的配方,使其能够支持气单胞菌属和 Plesiomonas spp 以及常见的肠杆菌科的生长。因此,它可以用作研究肠道疾病的通用介质。然而,为了提高其分离气单胞菌的性能,建议添加 5 mg/l 的氨苄青霉素。氨苄青霉素作为气单胞菌属的选择剂的有效性已被几位工作人员报告 2,3,4,5,6。气单胞菌培养基 (Ryan) 的值是氨苄青霉素的推荐水平远低于可抑制某些气单胞菌菌株7 的水平。

气单胞菌培养基 (Ryan) 的实用性及其优于其他一些用于检测气单胞菌属的公式。在自来水、瓶装水和包括肉类、家禽、鱼类和海鲜在内的食物中,已有报道8,9,10。气单胞菌培养基 (Ryan) 由 MAFF/DHS 食品微生物安全指导小组指定,用于检测和计数临床标本中的嗜水气单胞菌 11。气单胞菌广泛存在于土壤和水中,它们会导致鱼类和两栖动物的疾病。它们也存在于未经处理和氯化的饮用水、生食和生牛奶中11,12。人们认为气单胞菌属 spp12,13 引起胃肠道感染的主要原因是摄入受感染的水 14,15。

这些微生物在胃肠道疾病中的作用仍存在争议,但迅速扩大的文献表明气单胞菌可在成人和儿童中引起广泛的肠道症状 5,16在研究腹泻病时使用这种选择性培养基。

已开发出气单胞菌培养基基质以改进气单胞菌从临床和环境样本中的计数和分离。

技术
1. 按照说明准备培养基并倒入无菌盘中。制备的培养基可在 2-8°C 下储存长达 5 天。
2. 用食物、粪便等悬浮液接种平板,稀释后在接种平板上形成单个菌落。
3. 将板在 30-35°C 有氧培养 24 小时。如果需要进一步孵育,请保持在室温 (22-25°C)。
4. 检查板中是否存在中心较深的深绿色、不透明菌落。用生化测试确认身份。

The typical colonial appearance of Aeromonas isolates on this medium is as follows:

Aeromonas species: Dark green, opaque with darker centre, diameter 0.5-1.5mm.
Pseudomonas species: Blue/grey translucent, diameter from pinpoint to 0.25mm.

Storage conditions and Shelf life
Store the dehydrated medium at 10-30°C and use before the expiry date on the label.
Store the prepared plates of medium 2-8°C.

Appearance
Dehydrated medium: Straw coloured, free-flowing powder
Prepared medium: Green coloured gel

储存条件和保质期
将脱水培养基储存在 10-30°C 并在标签上的有效期之前使用。
将准备好的平板储存在 2-8°C 的培养基中。

外貌
脱水介质:稻草色、自由流动的粉末
制备培养基:绿色凝胶

Quality control

Positive control: Expected results
Aeromonas hydrophila ATCC® 7966* Good growth; opaque green colonies with dark centres
Negative control:
Escherichia coli ATCC® 11775* No growth
* This organism is available as a Culti-Loop®

Precautions
Although Aeromonas and Plesiomonas spp will grow on the medium if ampicillin is omitted, it will be more difficult to distinguish them from the other organisms present on the plate. Suspected colonies of Aeromonas spp must be confirmed by biochemical tests.

References
1. Ryan N. (1985) Personal communication.
2. Moulsdale M.T. (1983) The Lancet i: 351.
3. Rogol M., Sechter I., Grinberg L., Gerichter Ch. B. (1992) J. Med. Microbiol. 12. 229-231.
4. Richardson C.J.L., Robinson J.O., Wagener L.B. and Burke V. (1982) J. Antimicrob. Chemother. 9. 267-274.
5. Atkinson M. (1986) Culture Vol. 7, No.2.
6. Mishra S., Nair G.B., Bhadra R.K., Sikder S.N. and Pal. S.C. (1987) J. Clin. Microbiol. 25. 2040-2043.
7. Rahim Z., Sanyal S.C., Aziz K.M.S., Huq M.I. and Chowbury A.A. (1984) Appl. Environ. Microbiol. 48. 865-867.
8. Holmes P. and Sartory D.P. (1993) Letters in Applied Microbiol. 17. 58-60.
9. C. Pin M.L., Marin M.L., Garcia J et al (1994) Letters in Appl. Microbiol. 18. 190-192.
10. Warburton D.W., McCormick J.K. and Bowen B. (1994) Can. J. Microbiol. 40. 145-148.
11. Steering Group on the Microbiological Safety of Food (SGMSF). Methods for use in Microbiological Surveillance (1994) MAFF. Ergon House London SW1P 3TR.
12. Buchanan R.L. and Palumb S.A. (1985) J. Food Safety 7. 15-79.
13. Burke V., Robinson J., Gracey M., Peterson D. and Partridge K. (1984) Appl. Environ. Microbiol. 48. 361-366.
14. George W.L. (1987) Clin. Microbiol. Newsletter 9. 121-122.
15. Holmberg S.D., Schell W.L., Fanning G.R., Wachsmith L.K., Hickman-Brenner F.W., Blake P.A., Brenner D.J. and Farmer III J.J. (1986) Ann. Intern. Med. 105. 683-689.
16. Moyer N.P. (1987) J. Clin. Microbiol. 25. 2044-2048.

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