– Bacteria (at least four species, dominated by Pseudomonas putida, Bacillus
cereus, Acinetobacter sp., and coryneform bacteria)
Aeolosoma hemprichi, Philodina erythrophthalma, Lecane sp., Cyclidium glaucoma, Chlorella sp., Scenedesmus quadricauda, and Tolypothrix sp. must be
counted individually. However, the algal biomass can be substituted for chlorophyll
a and b. In the case of substitution of the algal biomass for chlorophyll a and b, it is
desirable to quantitatively measure phycocyanin, phycoerythrin, and
allophycocyanin.
6.1.1 Aeolosoma hemprichi and Philodina erythrophthalma
The number of Aeolosoma hemprichi and Philodina erythrophthalma should be
counted under an optical microscope using a glass slide with a grid as a borderline.
Independent measurements should be performed more than three times and with a
total of more than ten individuals in 5 mL or less. The unit of measure is N/mL.
6.1.2 Lecane sp. and Cyclidium glaucoma
A hemocytometer should be used. to count the number of Lecane sp. and Cyclidium
glaucoma with a phase contrast microscope. Independent measurements should be
performed more than three times and with more than 20 individuals in total in 300 μL
or less. The unit of measure is N/mL.
6.1.3 Chlorella sp. and Scenedesmus quadricauda
As Chlorella sp. and Scenedesmus quadricauda exist. as both single cells and
colonies, a hemocytometer should be used to count them with a phase contrast
microscope after disaggregating the colonies by sonication (20 s). Independent
measurements should be performed more than three times and measured until the
more abundant alga reaches 400 cells in total. The unit of measure is cells/mL.
6.1.4 Tolypothrix sp.
Because identifying species. of Conferva (filamentous algae) is difficult and the
measurement accuracy decreases slightly when sonicated, it is best not to sonicate it
before measurements are taken. Many Tolypothrix sp. become twisted, further
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K. Kakazu et al.
cereus, Acinetobacter sp., and coryneform bacteria)
Aeolosoma hemprichi, Philodina erythrophthalma, Lecane sp., Cyclidium glaucoma, Chlorella sp., Scenedesmus quadricauda, and Tolypothrix sp. must be
counted individually. However, the algal biomass can be substituted for chlorophyll
a and b. In the case of substitution of the algal biomass for chlorophyll a and b, it is
desirable to quantitatively measure phycocyanin, phycoerythrin, and
allophycocyanin.
6.1.1 Aeolosoma hemprichi and Philodina erythrophthalma
The number of Aeolosoma hemprichi and Philodina erythrophthalma should be
counted under an optical microscope using a glass slide with a grid as a borderline.
Independent measurements should be performed more than three times and with a
total of more than ten individuals in 5 mL or less. The unit of measure is N/mL.
6.1.2 Lecane sp. and Cyclidium glaucoma
A hemocytometer should be used. to count the number of Lecane sp. and Cyclidium
glaucoma with a phase contrast microscope. Independent measurements should be
performed more than three times and with more than 20 individuals in total in 300 μL
or less. The unit of measure is N/mL.
6.1.3 Chlorella sp. and Scenedesmus quadricauda
As Chlorella sp. and Scenedesmus quadricauda exist. as both single cells and
colonies, a hemocytometer should be used to count them with a phase contrast
microscope after disaggregating the colonies by sonication (20 s). Independent
measurements should be performed more than three times and measured until the
more abundant alga reaches 400 cells in total. The unit of measure is cells/mL.
6.1.4 Tolypothrix sp.
Because identifying species. of Conferva (filamentous algae) is difficult and the
measurement accuracy decreases slightly when sonicated, it is best not to sonicate it
before measurements are taken. Many Tolypothrix sp. become twisted, further
46
K. Kakazu et al.