Research Article
Viability of Antifungal Metabolite Producing Pseudomonas Bacteria

M.S. Shathele and A. Fadlelmula

Research Journal of Microbiology, 2009, 4(10), 361-365.

Abstract

The objectives of this study were to determine the suitability of transport medium (ice jells) and estimate the duration of viability of Pseudomonas in the transport medium. Bacteria of the genus Pseudomonas comprise a large group of the active biocontrol strains as a result of their general ability to produce a diverse array of potent antifungal metabolites. These include simple metabolites such as 2,4-diacetylphloroglucinol, phenazine-1-carboxylic acid and pyrrolnitrin [3-chloro-4-(2-nitro-3-chlorophenyl)-pyrrole], as well as the complex macrocyclic lactone, 2, 3-de-epoxy-2, 3-didehydro-rhizoxin. Pyrrolnitrin is active against Rhizoctonia sp., Fusarium sp. and other pathogenic fungi and it has been used as a lead structure in the development of a new phenylpyrrole fungicide. The survival rates of four different pseudomonad strains after continuous incubation for 4 h in the cold temperature (4°C) were: 94.8% for P. putida strain CBD, 94.5% for P. aeruginosa No. BRCH and 62.1% for Pseudomomas species (fluorescent) with lowest survival rate of 33.5% for P. aeruginosa strain H. Since, there were no drastic reductions in the survival rates, the study findings suggest that the transport medium would be generally suitable for these cold-sensitive bacteria.

ASCI-ID: 83-347

Table 1). The Pseudomonas aeruginosa strain BRCH count declined gradually from 9.1x108 at zero hour to 8.9x108 at 3 h and 8.6x108 at 4 h of incubation. The P. aeruginosa strain H showed marked decline in the colony count from 2x109 to 6.7x108 at the end of continuous incubation for 4 h. The P. putida strain CBD showed gradual decrease in the colony count from 5.8x108 to 5.6x108 at 2 h and further decreased to 5.5x108 after continuous incubation for 4 h. The Pseudomonas sp. (fluorescent) showed moderate decline in the colony count from 6.6x108 at 0 h, to 6.5x10°, 5.8x10°, 4.7x108 and 4.1x108 at 1, 2, 3 and 4 h incubation period, respectively. However, survival rates of four different pseudomonad strains after continuous incubation for 4 h in the cold temperature (4°C) were 94.8% for P. putida strain CBD (the highest survival rate), 94.5% for P. aeruginosa No. BRCH and 62.1% for Pseudomonas species (fluorescent). On the other hand, the lowest survival rate of 33.5% was for P. aeruginosa strain H.


Table 1: Viable cell count of four different strains of Pseudomonas sp. at dilution 10-6 of skim milk1
1Plating factor =10

Since there were no drastic reductions in the survival rate of different strains which suggests that the transport method would be suitable even for these cold-sensitive bacteria. The experimental findings were comparable to those reported by Johansson et al. (2003), who reported that the influence of environmental factors during isolation on the composition of potential biocontrol isolates is largely unknown. They identified Isolates in this group as Pseudomonas sp., they were fluorescent on King's medium B and had characteristic crystalline structures in their colonies. Members of this morphological group grow at 1.5°C and produce an antifungal polyketide (2, 3-deepoxy-2, 3-didehydrorhizoxin [DDR]).

However, the study results do not agree with those of Krieg and Holt (1984) and Wolf (1972) who stated that many Pseudomonas strains are not easily maintained at refrigerated temperature and relatively die within a short time at 4°C. Because many Pseudomonas strains are inactivated at low temperatures and are cold sensitive.

CONCLUSIONS

The viability of four different Pseudomonas strains was considerably affected by the incubation time. The survival rates of four different pseudomonad strains after continuous incubation for 4 h in the cold temperature (4°C) were 94.8% for P. putida strain CBD (the highest survival rate), 94.5% for P. aeruginosa No. BRCH and 62.1% for Pseudomonas species (fluorescent). However, the lowest survival rate of 33.5% was obtained for P. aeruginosa strain H. Since, there were no drastic reductions in the survival rate of different strains which suggests that the transport medium would be suitable even for these cold-sensitive bacteria.

All the strains gave more than 300 and less than 30 colony forming units at dilution 10-4, 10-5 and at dilution 10-7, respectively, when incubated up to 4 h in ice chest.

ACKNOWLEDGMENT

The authors gratefully appreciate Mr. Abdul Latif Alsagar for assistance and help in research.

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