Agar culture

Agar Culture Media fails to solidify. Insufficient quantity of agar or distribution thereof. Thoroughly mix media before pouring, 

Media boils out of vessel or flask containing it Excessive escape of steam from pressure cooker. Do not vent pressure cooker until reaching 1 psi. Crease pressure cooker seals with thin film of petroleum jelly, For pressure cookers using 5. 10. 1 5 lb weights, do not operate so steam escapes. 

Contamination oocurs in petri dishes after pouring media but before inoculation. High contaminant spore load in lab. Clean, paint lab. Install laminar flow hood. 

No growth from spores or tissue transferred. Wrong type of media. Wrong pH Old or dehydrated spores. Scalpel or loop too hot. Sugar in media caramelized. See media preparation. See media preparation Soak in sterilized water for 12-24 hours. Cool tool before contacting spores or tissue. Lower sterilization pressure and temp, to recommended levels. 

Contamination occurs around Inoculum (spores or tissue) Obtain cleaner spores or 

---

Algal cultures in exponential phase are used for inoculating Petri dishes containing agarized culture medium... 

Figure 12. A single hypha from an agar grown culture of C. versicolor showing labeling when the primary antisera was replaced with antisera to laccase from malt agar cultures of Agaricus bisporus. Magnification x 37,800. All bars, 1 pm. All tissue was unstained and consequently of low contrast.

Cotton swabs or RODAC plate (nutrient agar culture medium)... 

Agar Culture Medium Add 6.0 g of agar to 500 mL of Liquid Culture Medium, and heat with stirring on a steam bath until the agar dissolves. Add approximately 10-mL portions of the hot solution to test tubes, cover to prevent contamination, sterilize by heating in an autoclave at 121 ° for 15 min, and cool tubes in an upright position to keep color formation to a minimum. Store at 10° in the dark. 

Figure 21.1 Photomicrograph of Bacillus anthracis Uom an agar culture demonstrating spores Fuchsin-methylene blue spore stain. Anthrax CDC...

The two-layer soft agar culture system, originally introduced by Hamburger and Salmon (1980), consists of a bottom layer of hard agar, and a soft layer of agar in which cells are dispersed. 

As medium, a 4.5% aqueous malt extract solution having a pH of 5.4 is employed. One liter of this solution is sterilised in a 2-liter Erlenmeyer flask for 20 minutes at 110° C., and then inoculated with about 5 X10 (to the) 8 th) conidia of a 20-day old agar culture of the new strain and incubated for 3 days at 23° C. on a rotating shaking machine. A compact culture of fine mycelium flakes is formed. The flakes consist of loose clusters of hyphae and have a diameter of 2-4 mm. No alkaloids can be detected. 

This colony is broken up with a spatula into fractions which are transferred to a test tube with 12 cc. of the following agar culture medium ... 

Table 1. Increase of plating efficiency by CF and CF after water/diethyl ether repartition. Ten milliliters of agar culture medium were inoculated with 1 x 103 cell units/ml and were incubated for 20 days at 25 ° C. Mean value from three experiments and S.E. are reported. (Reproduced with permission from reference 16. Copyright 1987 Elsevier.)...

Agar culture media, except in vitro and in vivo Aggressions, except in vitro and in vivo... 

Figure 8 Plesiospora globosa, infected nematode with conidiophores and conidia developing in agar culture. (From Drechsler, 1971.)...

Figure 15 Haptocillium sphaerosporum, conidiophores and conidia from agar cultures of several isolates. (From Zare and Gams, 2001b.)... 

Agar culture medium actinomycete Bacillus mycoides Bacillus vulgatus... 

Gelatin or agar culture medium (zones of inhibition)... 

Agar cultures of the fungus Corticium caeruleum will only express the antibacterial cortalcerone (6, scheme 7) after activating treatment with toxic vapors or when being subjected to supraoptimal temperatures (136). 

Pyocyanine. 1 -Hydroxy-S-methylphenazini u m hydroxide inner salt. C,3HWNJ3 mol wt 210.23. C 74.27%, H 4.80%, N 13.32%, O 7.6]%. Produced by Pseudomonas aeruginosa (Ps. pyocyanea, Bacillus pyocyaneus). Isoln from broth cultures Wrede, Si rack. Z. Physiol Chem. 140, 1 (1924). Isoln from agar cultures Sohoental, Brit J, Exp Pathol. 22, 137 (1941). Synthesis by dimethyl sulfate meth-ylation of a -hydroxyphenazine and alkaline treatment of the resulting methosulfate Wrede, Strack, Z. Physio/, Chem. 181, 58 (1929) Ber. 62, 2051 (1929) Surrey, Org- Syn. 26, 86 (1946) coll, voL HI, 753 (1955). 

The formation of gas in lactose broth and the demonstration of non-spore-forming bacilli in the agar culture shall be considered a satisfactory completed test, demonstrating the presence of a member of the B. coli group. 

---