Measurement of chlorophylls a and b and bacteriochlorophyll a in organisms from hypereutrophic auxinic waters
- 66 Downloads
Sewage lagoons and wastewater ponds from industrialised swine and poultry farms are typically hypereutrophic, auxinic and dominated by purple non-sulphur bacteria and unicellular green algae both typically growing photoheterotrophically. To manage such ponds, it is essential to know the balance between oxygenic and anoxygenic photosynthesis. Typical spectrophotometric algorithms to estimate chlorophyll use 750 nm as a zero (A750 nm) but a 750 nm zero protocol is unsuitable where substantial amounts of bacteriochlorophylls are present. Algorithms were developed to estimate chlorophylls a and b (Chl a and Chl b) and bacteriochlorophyll a (BChl a) in solvent in ethanol, 7:2 acetone/ethanol and 90% acetone. The algorithms use an 850-nm absorbance zero (A850 nm) well outside the absorbance ranges of both chlorophylls and bacteriochlorophylls in solvent. There are many habitats where the presence of anoxygenic photosynthetic bacteria is unsuspected and so using a routine A750 nm zero effectively masks their presence and leads to underestimations of Chl a and Chl b. The in-solvent red peak of bacteriochlorophyll c is too close to that of Chl a for BChl c and Chl a to be resolved spectroscopically, but its presence can be easily identified from in vivo scans. The spectroscopic advantage of 90% acetone is negated by its poor quantitative extraction of pigments. Acetone/ethanol (7:2) is an excellent solvent spectroscopically and as an extractant.
KeywordsAbsorbance Anoxygenic photosynthetic bacteria Oxygenic photosynthesis Bacteriochlorophyll Chlorophyll Algorithms
The author wishes to thank Prince Songkla University—Phuket for providing facilities for the project. The co-operation of Phuket Integrated Waste Management (Wichit Sub-district, Mueang Phuket, District, Phuket 83000, Thailand) in encouraging this study and allowing us to collect sewage pond water samples is gratefully acknowledged. My Masters student, Piamsook Chandaravithoon assisted me in collecting the sewage pond samples. The author would like to give special thanks to Dr. Robert J Porra (CSIRO, Plant Industry, Canberra) for his valuable comments on the manuscript. Some of the data presented here was used as part of an oral presentation (Chandaravithoon et al. 2017: Photosynthesis in a sewage pond) at the recent 8th Asian Pacific Phycological Forum at the Pullman Hotel, Kuala Lumpur, Malaysia, 8–13 October 2017 (CS-12-2).
The project was partially funded by the Faculty of Technology and Environmental Science, Prince Songkla University-Phuket.
- Chandaravithoon P, Nakphet S, Ritchie RJ (2017) Photosynthesis in a sewage pond. 8th Asian Pacific Phycological Forum, Pullman Hotel, Kuala Lumpur, Malaysia, 8–13 October 2017 (Abstract CS-12-2, p 160)Google Scholar
- Falkowski PG, Raven JA (2007) Aquatic photosynthesis, 2nd edn. Princeton University Press, Princeton, NJ, USAGoogle Scholar
- Gitelson A, Stark R. Oron G, Dor I (1997) Monitoring of polluted water bodies by remote sensing. In: Remote sensing and geographic information systems for design and operation of water resources systems, proceedings of Rabat symposium S3, April 1997. IAHS Publ. No 242, pp 181–188Google Scholar
- Kiang NY, Siefert J, Govindjee, Blankenship RE (2007) Spectral signatures of photosynthesis. I. Review of earth organisms. Astrobiology 7:222–251Google Scholar
- Kim M-K, Harwood CS (1991) Regulation of benzoate-CoA ligase in Rhodopseudomonas palustris. FEMS Microbiol Lett 83:199–203Google Scholar
- Larimer FW, Chain P, Hauser L, Lamerdin J, Malfatti S, Do L, Land ML, Pelletier DA, Beatty JT, Lang AS, Tabita FR, Gibson JL, Hanson TE, Bobst C, Torres JLT, Peres C, Harrison FH, Gibson J, Harwood CS (2004) Complete genome sequence of the metabolically versatile photosynthetic bacterium Rhodopseudomonas palustris. Nature Biotech 22:55–61CrossRefGoogle Scholar
- Papineau D, Walker JJ, Mojzsis SJ, Pace NR (2005) Composition and structure of microbial communities from stromatolites of Hamelin Pool in Shark Bay, Western Australia. Appl Environ Microbiol 2005:4822–4832Google Scholar
- Porra RJ (1990) A simple method for extracting chlorophylls from the recalcitrant alga, Nannochloris atomus, without formation of spectroscopically-different magnesium-rhodochlorin derivatives. Biochm Biophys Acta 1019:137–141Google Scholar
- Porra RJ (1991) Recent advances and reassessments in chlorophyll extraction and assay procedures for terrestrial, aquatic and marine organisms, including recalcitrant algae. In: Scheer H (ed) Chlorophylls. CRC Press, Boca Raton, pp 31–57Google Scholar
- Porra RJ (2011) A proven simultaneous equation assay for chlorophyll a and b using aqueous acetone and similar assays for recalcitrant algae. In: Roy S, Llewellyn CA, Egeland ES, Johnsen G (eds) Phytoplankton pigments: characterisation, chemotaxonomy and applications in oceanography, SCOR-UNESCO 2011, Appendix 8A. Cambridge University Press, pp 366–371Google Scholar
- Sletten O, Singer RH (1971) Sulfur bacteria in red lagoons. J Water Pollut Control Fed 43:2118–2122Google Scholar
- Zhang D, Yang H, Huang Z, Zhang W, Liu S-J (2002) Rhodopseudomonas faecalis sp. nov., a phototrophic bacterium isolated from an anaerobic reactor that digests chicken faeces. Intl J Syst Evol Microbiol 52:2055–2060Google Scholar