ACHIEVEMENT OF ZERO EMISSIONS BY THE BIOCONVERSION OF FISHERY WASTES INTO FERTILIZER - Fertilizers: Components, Uses in Agriculture and Environmental Impacts - page 73

Red Seaweed
A fresh colony of B. alcalophilus from a nutrient agar plate was transferred with a
platinum loop to a 10-ml tube containing 5-ml autoclaved Porphyra culture medium (10 g l -1
P. yezoensis powder and 1 g l -1 of NH 4 Cl, pH 7.5). The Porphyra powder was purchased from
a local market and ground using a porcelain mortar and pestle. The final samples were sieved
to achieve particle size homogeneity using a 100-mesh sieve. The ground powder was boiled
for 40 s after treatment with 0.1% (v v -1 ) H 2 SO 4 at 121 °C for 15 min to increase its solubility.
After inoculation, the tube was incubated in a rotary shaking incubator at 30 °C and 180 rpm
for 3 d until cells reached a late-log phase. A 5% (v v -1 ) inoculum from this tube was then
used to inoculate 100 ml of the Porphyra culture medium in an 250 ml Erlenmyer flask, and
the flask was incubated for 2 d. Then, wet cells from the flask were collected by
centrifugation at 8000 rpm, and the harvested cells were used as an inoculum for the
biodegradation experiment.
Brown Seaweed
A fresh colony of M. oxydans from a nutrient agar plate was transferred with a platinum
loop to a 10-ml tube containing 5-ml autoclaved Laminaria culture medium (10 g l -1 L.
japonica powder, 0.1 g l -1 of MgSO 4 , 0.1 g l -1 of NaCl, 0.1 g l -1 of CaCl 2 , 2 g l -1 of (NH 4 ) 2 SO 4 ,
and 0.5 g l -1 of KH 2 PO 4 , pH 6.8). The Laminaria powder was purchased from a local market
and ground using a porcelain mortar and pestle. The final samples were sieved to achieve
particle size homogeneity using a 100-mesh sieve. The ground powder was boiled for 40 s
after treatment with 0.1% (v v -1 ) H 2 SO 4 at 121 °C for 15 min to increase its solubility. After
inoculation, the tube was incubated in a rotary shaking incubator at 30 °C and 180 rpm for 3 d
until cells reached a late-log phase. A 5% (v v -1 ) inoculum from this tube was then used to
inoculate 100 ml of the Laminaria culture medium in a 250 ml Erlenmyer flask, and the flask
was incubated for 2 d. Then, wet cells from the flask were collected by centrifugation at 8000
rpm, and the harvested cells were used as an inoculum for the biodegradation experiment.
Biodegradation of Seaweed
To characterize the degradation of green seaweed polysaccharides by B. licheniformis
TK3-Y, experiments were performed in a 1000 ml flask (with a 250 ml working volume). The
culture medium that was used for the degradation experiment was the same medium that was
used for the culture in tubes, as described above. The previously harvested cells were
proliferated for 12 h under the same culture conditions and used as an inoculum (0.5%, w v -1 )
for this experiment. After inoculation, the flask was incubated in a rotary shaking incubator at
50 °C and 180 rpm for 5 d.
To characterize the degradation of red seaweed polysaccharides by B. alcalophilus ,
experiments were performed in a 1000 ml flask (with a 250 ml working volume). The culture
medium that was used for the degradation experiment was the same medium that was used for
the culture in tubes, as described above. The previously harvested cells were proliferated for
12 h under the same culture conditions and used as an inoculum (0.5%, w v -1 ) for this
experiment. After inoculation, the flask was incubated in a rotary shaking incubator at 30 °C
and 180 rpm for 5 d.