The use of rice straw broth as an appropriate medium to isolate purple nonsulfur bacteria from paddy fields

The aims were to explore an appropriate isolating medium for obtaining purple nonsulfur bacteria (PNSB) for use as biofertilizers in saline paddy fields and to obtain pure cultures. We therefore chose a defined isolating medium containing 0.25 percent NaCl, (Glutamate-Acetate broth, GA) and a rice straw broth to compare them for numbers of PNSB obtained, time to obtain pure cultures, diversity and costs. A total of 30 water and 30 sediment samples were collected from saline paddy fields in southern Thailand and used to isolate PNSB in both the isolating media. Based on 60 samples and a period of 13 days incubation under anaerobic light conditions, a greater number of samples produced PNSB growth in GA broth after only day 3; however, after that the rice straw broth provided about a 2 fold increase in the number of samples that produced PNSB growth. Colonies isolated from GA broth required a significantly higher number of repeated streaking to obtain a pure culture (average 3.5) than those from rice straw broth (average 2.7) and the latter medium also produced significantly (P < 0.05) more isolates per sample. Sixty samples of water and sediment, from rice paddies with salinity (average, 3.43 +/- 0.67 mS/cm) and slight acidity (average, pH 5.84 +/- 0.42) provided 62 PNSB isolates by GA broth and 210 isolates by rice straw broth, and rice straw broth also produced a greater prevalence of PNSB. Estimates of the costs based on current prices of media, Gas Pak and electricity to obtain PNSB with the use of GA broth was roughly 6 times higher than for the rice straw broth.

Isolation of purple nonsulfur bacteria for the removal of heavy metals and sodium from contaminated shrimp ponds

In order to determine whether waters used for the shrimp cultivation contained toxic levels of heavy metals (HMs) and sodium (Na), analysis was carried out on 31 shrimp ponds in areas of southern Thailand. Purple nonsulfur bacteria (PNB) were also isolated from the same ponds to investigate if they could be used for bioremediation of the above contaminants. The highest HMs concentrations of the sediment samples in mg/kg dry weight were found as follows: 0.75 cadmium (Cd), 62.63 lead (Pb), 34.60 copper (Cu) and 58.50 zinc (Zn). However, all sediment samples met Hong Kong standards for dredged sediment. In contrast, contamination of Cu (9-30 ug/L) and Zn (140-530 ug/L) exceeding the standard guidelines for marine aquatic animal set by the Pollution Control Department, Thailand, were found in 32 and 61 percent of water samples, respectively. Two metal resistant PNB isolates, NW16 and KMS24, were selected from the 120 PNB isolates obtained. Both isolates reduced the levels of HMs by up to 39 percent for Pb, 20 percent for Cu, 7 percent for Cd, 5 percent for Zn and 31 percent for Na from water that contained the highest levels of HMs found and 3 percent NaCl when cultured with either microaerobic-light or aerobic-dark conditions. The isolate NW16 removed a greater percentage of the HMs than the isolate KMS24, but the isolate KMS24 was able to survive better under a greater variety of environmental conditions. Both strains were therefore suitable to use for further investigating their abilities to remediate water contaminated with HMs and Na.