Bio-gas and methane production from the microbial anaerobic decomposition of water hyacinths (Eichhornia crassipes) (Mart) Solms was investigated. These experiments demonstrated the ability of water hyacinths to produce an average of 13.9 ml of methane gas per gram of wet plant weight. This study revealed that sample preparation had no significant effect on bio-gas and/or methane production. Pollution of water hyacinths by two toxic heavy materials, nickel and cadmium, increased the rate of methane production from 51.8 ml/day for non-contaminated plants incubated at 36 C to 81.0 ml/day for Ni-Cd contaminated plants incubated at the same temperature. The methane content of bio-gas evolved from the anaerobic decomposition of Ni-Cd contaminated plants was 91.1 percent as compared to 69.2 percent methane content of bio-gas collected from the fermentation of non-contaminated plants

Gordon, J.

Mcdonald, R. C.

Wolverton, B. C.

English

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hAgA VL(CMM(CAL	 TAI-X 72725
N2NDNANDUM
	 JULY, 1915
(4A,S.A-Th - X-72725)	 BIO-CONVEP. SION OF WATF,R
HYACINTHS INTI MFTHANE GAS, PA:; T 1 (NASA)
13 p IW 3;3.25	 CSC  10A
N75-275b4.
U tic I a.;
G3/44 2b7b6
B I O-CONVERS ION OF WATER HYAC I NTHS
I NTO METHANE GAS: PA RT
By B. C. Wolverton
R. C. McDonald
J. Go rdon
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NASA
NATIONAL SPACE TECHNOLOGY LABORATORIES
BAY ST. LOUIS, MISSISSIPPI 39520
N5TL FORM 17 (JAN 1979)
i	
1
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I. Re PORT NO.
	
-[777.7­vW04NPAIrN'V ACCW%%II.N NO. 1,	 NRCIPIFNT 1 CATALOG N0.
'I'M-X-72725
4.	 NIPORT DATI[
.,	 TITLE ANO SUOTITL[
IIIo-Converslon of Water Hyacinths Into Methane Gan:	 Part 1 Jul	 1975
•,	 Pf N/OHMING T)fK.ANIZATION COOT
7.	 AUTNON(S) ^.	 PIN P ORMING ORGANIZATION
H. C. Wolverton, It. C. McDonald, .1. Gordon
N[ PONT NO.
9.	 Pr Rf ORMING ORGANIZATION NAM[ AND ADDMCSG I1).	 WORK UNIT NO.
Natlon:Il Space Technology Laboratories
I1.	 CONTRAC'r OR GRANT NO,Bay St.	 L.cxlls, Mississippi 39520
17.	 1PON-%oRINc- AGr NCV NAME AND ADDRF'I4 11,	 TVPF OF NFPORT % R^RIOD
C(JVFHrr)
National Aeronautics Ind Space Administration Technical Memorandum
Washington, D. C. 20546 14,	 SPON SO NING A l .FNCV con y
11,	 `, Ur. Pl. rMrNTARY NOTrS
lL!',ivrgy
tez,im program tests results, NASA, Office of Applications sponsored prograrr -
TOP 644-02-02) Vascular Aquatic Plants for Pollution Control and Sources of
  ;Ind FOud.
1F,	 Ali S TRACT
IBio-gas and methane production from the microbial anaerobic decomposition of water hyacinth.,
(EicW --nia crassipes) (Mart) Solms was I rive sti gated.	 'These experiments demonstrated the
ability of water hyacinths to produce an average of 1:3. 1) m) of r;leth:lne gas ner gram of wet
plant weight.	 This study revealed that sample preparation had no significant effect on hio - gas
and/or methane production.	 Pollution of water hyacinths by two toxic heavy materials, nickel
:Intl cadmium, Increased the rate of methane production from 51. 8 ml/day for non-contaminated
plants Incubated at 36°C to 81. 0 ml/day for NI-Cd contaminated plants incubated at the same.
temperature.	 The methane content of blo-ga g evolved from the anaerobic decomposition of N1-Cd
contaminated plan. , wa ,s 91. 1 percent as compared to 69. 2 percent methane content of bio - gas
collected from the fermentation of non-contaminated plants.
Use of trade names or names of manufacturers in this report does not constitute an official
endorsement of such products or manufacturers, either expressed ur implied, by the National
Aeronautics and Space Administration or any other agency of the United States Gover.-.ment.
17,	 KFY WORDS	 --	 Is,	 DISTRIBUTION 5"1"ATr MrNT
Bio- gas	 Unclassified - Unlimited
Methane
Anaerobic fermentation
19,	 SECURITY CLASSIF,(af thl7 r@path
	 20.	 SECURITY CLASSIF. (ef Iwl• Grp) 	 21,	 NO. OF PAG<r	 t122
Unclassified	 Unclassified	 13	 N11 iti
s
NSTI_ FORM 13 i. J^N 1977)
T1
G	 TECliivfCAl. MEMORANDUM X-72725
Ei I O-CONVER S i ON OF WA f ER HYACINTHS
INTO METHANE GAS: PART I
Two of the most pressing problems facing; the United Stater and other industrial
nations today are rapid depletion of vital natural resources and Ixf)lution of the environ-
ment. One important tractor in the rise of the United States to its present high indus-
trial level has been alp abundance of fossil fuel resources. Presently, available coal,
oil, and large reservoirs of underground natural gas are all produced through natural
decomposition of prehistoric forms of life. Modern society is depleting these resources
at an alarming rate, and renewable sources must be developed within the near future fo.-
continued industrial growth.
	 }
As we deplete our natural rese-irces, we are also polluting our environment at
the same alarming rate. Fortunately, a large number of the minerals with which we
are polluting our water systems have the potential of being recovered through natural
biological processes.
Recently, the ability of vascular aquatic plants to remove organic chemicals,
heavy metals, pesticides, and nutrients from polluted waters has been demonstrated
(1, 2, 3, 4, 5, 6). Harvested plant material from these experiments is a potential
source of renewable resources, such as natural gas, fertilizers, and other valuable
minerals. Vegetation can be fermented anaerobically and made to release bio-gas
containing a high percentage of methane (7, 8).
Many factors can affect the actual amount of gas and fertilizer produced from
the digestion of plant material. One of the most important of these factors is the car-
bon to nitrogen (C/N) ratios of the material used. For maximum bio-gas production
the C/N ratin should be approximately 30:1.
Water hyacinths (Eichhorni2 crasj s (Mart.) Solms, were c ,,sell for this study
because they have demonstrated the most promise in removing chemicals from polluted
waterF and producing large quantities of harvestable plant matt r'ui possessing a de-
sirable C/N ratio for maximum methane gas production. This aquatic plant has the
potential of producing over 240 Kg (529 lbs.) of dry plani material per 0..'0 hectare
(acre) per day while removing undesirable chemicals from waste waters.
MATERIALS AND METHODS
Water hyacinths used in these experiments were grown by vegetative reproduc-
tion inside a greenhouse maintained between 25°C and 10°C. Several plants were
selected for each experiment whose total wet mass ranged from 3008 to 87bg. In one
of the fermentation studies, water hyacinths were contaminated with nickel and cadmium
by exposing them to a known concentration of cadmium and nickel prior to fermentation.
The 542 3or wet mass of water hyacinths absorbed 5. to mg of nickel and ti. 87 mg of
c • admim, fruni 2. 5 liters of NI-Cd contnmin.,ted distilled water. Metal concentration
was monitored by atomic absorption.
For four of the five fermentation units, the plants were chopped into approxi-
mately one-inch lone; pieces. Water hyacinths were blended into a slurry form for the
other fermentation study. The chopped or blended water hyacinths were transferred
into three liter Erlenmeyer flasks covered with aluminum foil to prevent exposure to
light.
Starter seed for the fermentation studies was prepared by allowing water hya-
rinrhs to decompose under water -nd mud approximately six months in an anaerobic
condition. For each fermentation unit incubated at 36°C, approximately 26 g of this
seed was blended with ;150 nil of distilled water. Fifty grams of seed and 800 ml of
distilled water were used for each experiment at room temperature. The sediment
from the seed and water mixtures was allowed to settle 30 minutes, and the supernatant
liquid was then decanted into each flask containing water hyacinths.
The Erlenmeyer flasks were sealed to the atmosphere with two-hole rubber
stcjppers. One outlet was fitted with a rubber septum for gas chromatographic sam-
pling, and the other outlet was connected with rubber hose to a sealed co:atainer filled
with water acidified with sulfuric acid. The displacement of water in tfe second con-
tainer by the bio-gas produced in the fermentation flask pro%-ided a convenient method
of monitoring the volume of bio-gas production. Mixing of the water hyacinths was
accomplished by shaking the fermentation flask once each day.
Samples for gas chromatographic analysis were taken thronigh the rubber septum.
Matheson Gas Products c.p. gl:..4de methane was used as the methane standard. The
methane content of the bio-gas was analyzed by gas chromatography using a Varian
2100 GC with a flame ionization detector. Gas chromatographic conditions were:
Column:	 6' x 1/4', ii.d. glass
Packing:	 Porapak`='Q 150-200 mesh
2
rFlow Itates:
	 Nitrogen 60, Ilydrogen :15, and
ml/mimile	 Air 235
Temperature:	 Detector 155°t', injection 150`C,
Colunln 55•C
Carrier Gas:	 Nitrogen
Sample Size:
	
5 u I and 10 u 1
RESULTS AND D I SCU`_ 310N
Five laboratory experiment:; were conducted in order to evaluate the effect of
temperature, toxic metal contamination, and plant prepar;,tion on the production of
bio-gas and/or methane from the y microbial anaeroble decomposition of water h^ , a-
cinths, Eichho -nia crassipes) (Hart. ) 3o ► lnis.
Two of the three experiments were conducted at room temperature (25 60 _ 50C)
and contained wa er hyacinths chopped into one-inch long pieces. 'I'll(- other experi-
ment was conducted under the same conditi-is except the water hyacinths were blended
into a slurry form. According to the data presented in Tables 1 and 2, there was no
significant difference in the results of these three experiments. The chopped water
hyacinths produced 11.0 and 6.4 nil methane per gram wee weight, as compared to a
Auction of 7.9 ml methane per gram weight for the blended water hyacinths. The
methane content of the total bio-gas pro tluced by the to aerobic decomposition of the
slurried water hyacinths was 61. 1`%.
 This value was comparable to the 57.2` -, and
61. 57 methane content of the bio-gas produced in the other two experiments with chop-
ped water hyacinths at room temperature.
Temperahtre played an important role in the rate of bfo-gas and methane produc-
tion. The time lag between the production of bio-gas and the production of methane gas
was reduced from an average of eight days for those maintained at 25 00 ► 5 °(' to approx-
imately one day for experiment 4 incubated 3VC. The methane content of the total bio-
gas produced in experiment 4 a' 36°C was 69.2`l;. This percent methane was higher
than the average methane content of 59.97, for the three experiments conducted at room
temperature.
Comparison of the data for experiments 4 and 5 in Table l in which chopped plants
were incubated at 36°C showed that nickel and cadmium contamination of the I,3 ater hya-
cinths at concentration levels of 9.95 and 12.66 mg/kg wet weight nickel ant,, cadmium,
respectively, had no adverse effect on the percent methane content, volume of methane
produced per unit wet weight, or the rate of bio-gas and /nor methane production. In
fact, the Ni-Cd contaminated plants produced bio-gas with a 91.17 methare content,
as compared to the lower value of 69.27 for the other experiment incubated at 36°C.
:3
4The total volume of blo-gas produced from the Ni-Cd contaminated plants was less than
the volume produced from the non-contaminated plants; however, clue to the much higher
percentage of methane in the bio-gas, the Ni-Cd ccurtaminated plants yielded 11.3 ml
methane per gram wet weight, as compared to only 8.9 ml methane per gram
wet weight for the nun-contaminated plants. A, so, the average rates of bio -gas and
methane production (87.5 and 81.0 ml/day, respectively) for the Ni-Cd contaminated
water hyacinths for the first 65 days r;f incubation was significartly higher than the
rates for the non-contaminated plants (76.8 and 51. M ml/daY hi( -gas and mcthane pro-
duction, respectively) incubated at the same temperature.
CONCLUSION
This study on the anaerobic decomposition of water hyacinths revealed that sample
preparation, either chopped or blended, had no sitmificant effect on bio-gas and/or
methane production. Incubation of the experimental units at 36°C increased not only the
rate of bio-gas production, but also the methane content of the total bio-gas produced
in these experiments. Pollution of the water h y acinths by two toxic heavy metals,
nickel and cadmium, actually increased the rate of methane production ani
the methane content of the bio-gas evolved in the anaerobic decomposition
taminated plants.
Further studies are planned in this area utilizing the preliminary irnl
from these experiments. Temperature controlled, continuous fc( . fermei
bers with effiL,ent stirring devices are presently being dcvelol -t ',y NASA
National Space Technology laboratories.
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Table 2. Calculated Data of Blo-Kart tuul
Methane Production for Experiments 1 -5
Experiment M
1 , Methane
in Total I3fo- Ana
ml Di or-gas
per Gram
Wet Weight
nil Methane
per tram
Wet Wei &ht
1 61.5 17.9 11.0
2 57.2 11.1 6.4
3 61.1 12.9 7.9
4 69.2 12.9 ti. 9
11.35 91.1 12.4

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1 1
1:_'
1
fain-Ran
— _ — — •- Methane
10	 20	 30	 40	 50	 60	 70	 RO	 00	 100	 11 1,	 20	 130	 140
Days
Figure 2. Cumulative volume of bio-gas and methane gas per 1.0 Kg wet muss of chopped
water hyacinths at 25°C + 5* C versus numlxer of days elapsed since initiation of
anaerobic fermentation.
8
l .1
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i
^a
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10	 20	 30	 40	 50	 60	 70	 90	 90	 100	 1 1 f)	 1-10	 1;11r	 1 ;n
Days
nnure 3. Cumulative volume of hio-gas and methane g;fti per 1. 0 Kg wet :n,r„ of blended
water hyacinth; at 25*C + YC versus number of clays elapsed singe initiation of
anaerobic fermentation.
1-
lu
v
ti
W
0
L
E
7
C
J
f
10	 20	 30	 40	 50	 60	 70	 No	 90
	 100	 110	 120	 130	 140
Days
Figure 4. Cumulative volume of blo-gas :ind methane gas per 1.0 Kg wet mass of chopped
water hyacinths Incubated at 36°C versus number of days elapsed since initiation
of anaerobic fermentation.
10
------------
60	 7V	 MO	 4U	 JUO	 110	 120	 1311
Days
10	 20	 30	 40
I I
tr
E
O
E
I 'MWOOMMUMM Ow
Figure 5. CIII11LII:ItiVe volume of bio-gas and methane gas per 1.0 Kg wet mass of chopped
water hyacinths Incubated at :36*C versus number of (lays elapsed since initiation
of anaeroble fermentation.
REFERENCES
1. Wolverton, B. C. and D. D. Harrison 1975. i.- t"atic Plants for Removal of
Mevinphos from the Aquatic Environment. (In Press) Journal of Mississippi
Academy of Science.
Wolverton, B. C. ami M. M. McKown 1975. Water Hyacinths for Removal of
Phenol frorn Polluted Waters (In Heview), A ( ILI (' Rotany Jouriml, Amsterdam,
The Netherlands.
3. Wolverton, 11. C. 1975. Water Hyacinths for Removal of Cadmium and Nickel
from Polluted Waters. NASA Technical Memm-andum ';'bl-X-72721.
4. lA olverton, B. C. and R. C. McDonald 1975. Water Hyacinths and .Iliotor
Weeds fo-- Removal of Iead and Mercury from Polluted Waters. NASA Technical
Memorandum TM-X-"2723.
5. Rogers, H. H. and D. F. Davis 1972. Nutrient Removal by Water liyacintk.s,
Weed Se.: 20: 423-427.
6. Haller, W. T. and D. T. Sutton 1973. Effect of p1l and High Phosphorus Concen-
trations on Growth of Water hyacinth. 11 y a. C ►^ntr(d J. 11:"'i t^l
7. Muthant Digesters for Fuel Gas and Fertilizer 197:3. New Alchemy Institute,
Newsletter No. 3. Box 432, Woods Hole, Mass.
S.	 Singh, It. B. 1971. Bio-Gas Plant (venerating Methane ► rom Organic Wastes.
Gobar Ga. Research Station, Ajitmal, Etawah (U.P.) India.
i
12
APPROVAL
B 10-CONVERS ION OF WATER HYAC I NTHS
INTO METHANE GAS: PART 1
By H. u. Wolverton
It. C. McDonald
J. Gordon
The information in this report has been reviewed for security classification. Re-
view of any information concerning; Dapartment of Defense of Atomic Energy Commis-
sion programs has been made by the NSTI. Security Classification ( )fficer. This
report, in its entiret y . has been determined to be unclassified.
This document bas also been reviewed and approved for technical accuracy.
HENRY F. AUTF.R
Director, Applications Engrineering
National Space Tectmology Laboratories
1
13


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