African Journal of Biotechnology Vol. 7 (10), pp.

1516-1525, 16 May, 2008

Available online at http://www.academicjournals.org/AJB
ISSN 1684–5315 © 2008 Academic Journals

Full Length Research Paper

Compost bioremediation of hydrocarbon-contaminated

soil inoculated with organic manure
Harrison Ifeanyichukwu Atagana
Institute for Science and Technology Education, University of South Africa, Pretoria, South Africa.
E-mail: [email protected].
Accepted 10 April, 2008
-1
Contaminated soil (FAO: Lithosol) containing >380 000 mg kg total petroleum hydrocarbons (TPH) was
bioremediated by composting. The soil was inoculated with sewage sludge and incubated for 19
months. The soil was mixed in a ratio of 1:1 (v/v) with wood chips. The soil-wood chips mixture was
then mixed in a ratio of 4:1 with sewage sludge. Compost heaps were set up in triplicates on wood
pallets covered with double layers of nylon straw sheets. Control experiments which contained the
contaminated soil and wood chips but without sewage sludge were set up in triplicate. Moisture,
temperature, pH, ash content, C:N ratio of the compost mixture and TPH of the soil was monitored
monthly. The concentrations of selected hydrocarbons in the contaminated soil were measured
monthly during the incubation period. Temperature rose to about 58°C in the sewage sludge compost
within two months of incubation, while temperature in the control fluctuated between 15 and 35°C
throughout the incubation period. Total petroleum hydrocarbons (TPH) was reduced by 17% in the
control experiments and 99% in the sewage sludge compost at the end of the incubation period. The
concentrations of most of the selected hydrocarbon components were reduced by up to 100% within
the same period. Microbial activities were shown to correlate with the reduction in hydrocarbon
contents of the soil.

Key words: Bioremediation, composting, PAHs, sewage sludge, soil.

INTRODUCTION
-1 -1
The use of composting in bioremediation has received mg kg , chrysene 93.3 mg kg and benzo (a) pyrene
-1
little attention (Potter et al., 1999), despite its application 68.4 mg kg . These high concentrations of hydrocarbons,
in the treatment of soils contaminated with organic com- particularly the PAHs provided a good opportunity to
pounds for many years. Much of the work on treatment of study and further understand the potentials of compost
contaminated soils by composting (Valo and Slakinoja- bioremediation of organic compounds. Earlier studies
Salonen, 1986; Potter et al., 1999; Reid et al., 1999) has showed that higher molecular weight PAHs remained in
been done on soils with lower concentrations of the the soil after 16 weeks and 11 months of pilot-scale and
contaminating substances than used in the present study, full-scale land farming, respectively (Atagana, 2003;
in spite of the fact that composts have been reported to Atagana, 2004a). Earlier composting experiments using
have potential for remediation of heavily contaminated hydrocarbon-contaminated soil co-composted with cow
sites (Reid et al., 1999; Garcia-Gomez et al., 2003; manure and mixed vegetable waste showed that more
Manios et al., 2006; Marin et al., 2006). than 90% of the hydrocarbons including some of the
The soil used for this study was a lithosol (FAO) con- recalcitrant components were removed (Atagana et al.,
-1
taining >380 000 mg kg total petroleum hydrocarbons 2003). Co-composting hydrocarbon-contaminated soil
(TPH). Selected individual hydrocarbons present in the with poultry manure showed that PAHs could be removed
-1
contamination include: phenol 127 mg kg , o-cresol 15 from the soil by composting (Atagana, 2004b). This study
-1 -1 -1
mg kg , m-cresol 26 mg kg , p-cresol 23 mg kg , intended to exploit the high temperatures achieved during
-1 -1
naphthalene 158 mg kg , anthrecene 72.3 mg kg , composting, the high nutrient contents and the heavy
-1 -1
phenanthrene 256 mg kg , fluorene 68.3 mg kg , pyrrole microbial load of the sewage sludge in enhancing the
-1 -1
77.3 mg kg , pyrene 182.1 mg kg , fluoranthene 188.5 microbial removal of high concentrations of hydrocar-
 Atagana 1517

bons, particularly those of high molecular weight from the (Holt, 1994; MacFaddin, 1980). Fungal isolates were identified by
contaminated soil under investigation. microscopic examination with reference to Barnett and Hunter
(1972), Raper and Thom (1968).
The aim of this study was to investigate the effects of Analysis of variance (ANOVA) was done to determine the level of
inoculating soil heavily contaminated with a complex significance at p <0.05 between the results obtained at each period
mixture of hydrocarbons with sewage sludge on the of the composting process. All statistical analysis was performed
removal of hydrocarbons in a static-pile compost system. using PlotIT software.
The temperature regimes in the compost systems, the
changes in nutrient composition and moisture content
during the treatment period were studied. This was meant RESULTS AND DISCUSSION
to determine the requirements of the compost type and
its practical application for large-scale treatment of Changes in pH and temperature during composting
hydrocarbon-contaminated soils. The remediation target
-1
set in this experiment was 1 mg kg of the selected The pH of the sewage sludge compost and the control
-1
hydrocarbons and 1000 mg kg of total petroleum increased from 7.0 and 6.9 to 8.1 and 7.7, respectively
hydrocarbons (TPH) in the soil. The chosen targets were within four months of incubation and then fluctuated
informed by earlier targets set by government for similar between 6.2 and 7.7, 7.1 and 6.7, respectively (Figure 1).
hydrocarbons in soil and partly to the standards used in The pH ranges observed in these experiments are well
an earlier experiment (Lees, 1996) dealing with similar within the recommended range for composting organic
hydrocarbons. materials (Feinstein et al., 1986; Kubota and Nakasaki,
1991; Marin et al., 2006). The high increases in the first
MATERIALS AND METHODS
four months could be from high metabolic activities
possibly resulting in the production of intermediate meta-
Contaminated soil (FAO: lithosol) containing >380 000 mg kg-1 bolites in the compost systems. The decreases observed
hydrocarbons was mixed with wood chips in a ratio of 1:1 (v/v) to in subsequent months are attributed to the degradation of
improve aeration and then mixed with sewage sludge in a ratio of the compost and the hydrocarbons, which may have
4:1 (contaminated soil + wood chips : sewage sludge) (v/v). Tripli-
cate static pile compost heaps (about 350 kg each) were set up on
resulted in the release of acidic intermediate and final
wooden pallets that were covered with nylon fibre sheet in the open products that probably lowered pH of the mixture
yard. The compost heap was then covered with hay for insulation (Alexander, 1999; Eweis et al., 1999) (Figure 1).
and incubated for a total of 19 months. The control was a mixture of The temperature of the control experiment, which rang-
contaminated soil and wood chips without sewage sludge. ed between 12 and 30°C fluctuated with the daily diurnal
A temperature data logger with thermocouples located in the air temperatures, which ranged between 17 and 33°C
middle of the compost heaps was used to monitor changes in
temperature. The moisture content of the compost was measured
during the experimental period (Figure 2). Temperatures
weekly by using the method described by Forster (1995a) and in the sewage sludge compost rose to 58°C in the second
water was added as required. The pH of the aqueous extract of the month of incubation and remained relatively stable until it
compost mixture was measured at monthly intervals in triplicate started to decrease at the end of the third month reaching
with a pH meter (Crison Micro pH 2000TM). The ash content of the 30°C. Temperatures became relatively stable after the
compost mixture was determined at the start and at the end of the seventh month, fluctuating between 29 and 40°C for the
experiment by heating 10 g of the mixture in a furnace at 400°C for
6 h. remainder of the experiment (Figure 2). The large in-
CO2 evolution measured by using the closed jar method (Alef, crease in temperatures in the first two months was due to
7
1995) at room temperature was used to estimate microbial activity. the high initial microbial load, 1.28 x 10 in the sewage
Microbial plate counts of samples taken from three different levels sludge, which resulted in high metabolic activities (Figure
(20, 35 and 50 cm) in the compost heaps were carried out on 3). However, the high temperature became inhibitory to
nutrient agar and the data represented as colony forming units per
gram (cfu g-1). Total organic carbon was determined by a conduc-
continued microbial growth after two months, resulting in
tometric method (Forster, 1995b) extractable phosphorus by the a decrease in microbial activity (Figure 3). A subsequent
Bray-1-P test (Recommended Chemical Soil Test Procedures decrease in temperature in the fourth month (Figure 2)
No221, University of Missouri Agricultural Experiment Station, resulted in increase in microbial activity in the following
Columbia, Missouri 1998) and total nitrogen was determined by soil month. This phenomenon has been previously reported
digestion method (Forster, 1995c). by Potter et al. (1999) (Figures 2 and 3).
Total petroleum hydrocarbon (TPH) was determined by infrared
using US EPA Method No. 8440 (1996) and absorbance was
determined with a Nicolet Avatar 320 IR Spectrophotometer at
wave numbers between 2760 and 3070 cm-1 and an integration Changes in the C:N ratio of the compost during
value for the absorbance peak was automatically generated. composting
Changes in the concentrations of selected hydrocarbons were
determined by Soxhlet extraction and GC/FID analysis. The GC The sewage sludge and the contaminated soil had initial
was a Varian-3800 with argon as the carrier gas and fitted with a 3 C:N ratios of 9:1 and 306:1, respectively. The high C:N
m capillary column with 0.25 µm film thickness. Two temperature
programmes were used (Eriksson et al., 2000) in order to obtain
ration of the soil is attributed to the TPH content (>380
-1
good separation and quantification of the more volatile compounds. 000 mg kg ) of the soil, which will naturally inhibit
Identification of bacterial isolates was done by biochemical tests microbial growth (Baker and Herson, 1994: Alexander,
1518 Afr. J. Biotechnol.

8.5

7.5
pH of compost

6.5

6
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19
Time (Months)

Control Sewage Sludge

Figure 1. Changes in the pH of compost during composting. Values are means of three
replicates ± 1 Standard Error.

70

60

50
Temperature (OC)

40

30

20

10

0
0 2 4 6 8 10 12 14 16 18 20 22 24 26 28 30 32 34 36 38 40 42 44 46 48 50 52 54 56 58 60 62 64 66 68 70 72 74 76
Time (Weeks)

Air Temp. Temp. of Control Temp. of sewage sludge

Figure 2. Changes in temperature of the compost during composting. Values are means of three
replicates ± 1 Standard Error.

1999). The C:N ratio of the sewage sludge compost (between 25:1 to 35:1) (Anderson, 1991; Kubota and
mixtures was 23:1 (Table 1) at the start of the com- Nakasaki, 1991), they were considered adequate for this
posting. Although this nitrogen content is higher than experiment, considering the level of contamination in the
those recommended for effective compost bioremediation soil and the results obtained in earlier experiment in
 Atagana 1519

5000

4500

4000

3500
CO2 (ug)/dwt/day

3000

2500

2000

1500

1000

500

0
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19
Time (months)

Control Sewage sludge

Figure 3. Changes in the respiration rates of compost inhabiting microorganisms during composting. Values are
means of three replicates ± 1 Standard Error.

Table 1. Changes in the C:N ratios of compost mixture during incubation.

Treatment 0 6 Months 12 Months 18 Months

Contaminated soil (control experiment) 306:1 326:1 322:1 289:1
Sewage sludge compost 23:1 15:1 15:1 17: 1
Values are means of three ± 1 Standard Deviation.

which up to 90% of the total hydrocarbons were removed and 10% in the control. Sewage sludge has been repor-
(Atagana, 2003; Atagana, 2004a). The C:N ratio in the ted to enhance the degradation of hydrocarbons in soil-
compost and control changed as the incubation pro- compost mixtures (Hill and McCarthy, 1967; Wilson et al.,
gressed (Table 1). The decrease in nitrogen content in 1983). The rapid degradation of hydrocarbons in the
the sewage sludge compost was faster than in the control compost system was expected since sewage sludge is
probably due to higher microbial activity and higher rate rich in nutrients and has high microbial population
of breakdown of the hydrocarbon substrates in the (Combs et al., 2001; Schmitt and Rehn, 2002). At the end
compost systems. of the incubation period TPH had decreased by 17% in
Ash components of the compost mixtures changed the control experiment and 99.8% in the sewage sludge.
slightly from 4.62 to 4.60 in the sewage sludge compost The organisms growing on the nutrients present in the
and 6.37 to 6.41 in the control, which indicates that there compost system readily metabolise the contaminant
was no significant change in the mineral components of hydrocarbons in the compost mixture while still growing
the soil at the end of the experiment (Table 1). on the sludge. This deduction was made, as isolates from
the sewage sludge could not establish easily on the same
contaminated soil in the absence of the sewage sludge
Changes in total petroleum hydrocarbon (TPH) (results not shown). The organisms, while growing on the
content and microbial populations sludge substrate, probably produced enzymes that were
used in metabolising the hydrocarbons in the compost
The changes in TPH content of the sewage sludge com- matrix (Sutherland et al., 1995; Bardos et al., 1996; Diaz
post were significantly different at p<0.05 from the control et al., 1996). The high microbial load in the compost
experiment (Figure 4). In the first two months, decreases (Figure 5) at the start of the composting afforded the
in TPH levels were 67.8% in the sewage sludge compost population the opportunity to remain high while adapting
1520 Afr. J. Biotechnol.

450000

400000

350000
Total petroleum hydrocarbons (TPH) (mg kg )
-1

300000

250000

200000

150000

100000

50000

0
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19
Time (Months)

Control Sewage Sludge

Figure 4. Monthly reduction in total petroleum hydrocarbons (TPH) during composting. Values are means of
three replicates ± 1 Standard Error.

to and attacking the hydrocarbon substrate. Changes in the concentrations of selected

During the second month of the experiment, counts of hydrocarbons
7
heterotrophic micro-organisms increased from 1.28 x 10
7
to 1.86 x 10 in the sewage sludge, probably due to the The tested phenols (phenol, o-cresol, m-cresol and p-
ready supply of available nutrients in the sludge, and 3.78 cresol) and the 2- and 3- ring PAHs (naphthalene,
4 4
x 10 to 7.15 x 10 in the control (Figure 5). During the anthracene, phenanthrene, fluorene and pyrrole) were
-1
same period, the largest amounts of reduction in TPH removed below the remediation target of 1 mg kg by
content were observed (Figure 4). between the third and ninth month of incubation (Figures
Increases in microbial populations and rapid reductions 6 and 7). The removal of the 4- and 5- ring PAHs
in hydrocarbon content continued after the fourth month (pyrene, chrysene, fluoranthene and benzo (a) pyrene)
probably due to the decrease in temperature as men- became slower from about the eighth month, and residual
tioned earlier, and a decrease in nitrogen levels caused concentrations of chrysene continued to be above 1 mg
-1
by the initial high microbial activity (Piccinini et al., 1996; kg until the sixteenth month (Figure 8). A decrease in
Atagana, 2004b). The populations reached a maximum of temperature in the compost mixtures in the fifth month
7
3.3 x 10 in sewage sludge compost in the fourth month (Figure 2) resulted in an increase in microbial populations
4
and 7.6 x 10 in the control experiment in the eleventh (Figure 5) with a subsequent increase in the removal of
month before decreasing (Figure 5), probably as a result pyrene, chrysene and fluoranthene. The increased
of the large decrease in the concentration of hydrocar- decrease in concentration only became obvious for
bons and the nutrient base. The high temperatures benezo (a) pyrene by the seventh month (Figure 8). Apart
generated in the compost (Figure 2), which resulted in from the phenolics and naphthalene, most components
the increased loss of water from the soil-compost matrix persisted to the end of the incubation period in the control
may also have resulted in the decrease in microbial (Figures 6, 7 and 8).
population after the sixth month (Fan and Tafuri, 1994). Respiration of micro-organisms in the compost systems
Regular watering when necessary kept the temperature (Figure 3) and counts of heterotrophic microorganisms
from increasing higher than 58°C in the compost system (Figure 5) shows that microbial activity correlated with
(Figure 5). changes in TPH and selected hydrocarbons levels. The
 Atagana 1521

40000000

35000000

30000000
Counts of microorganisms (cfu g-1)

25000000

20000000

15000000

10000000

5000000

0
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19
Time (months)

Control Sewage

Figure 5. Counts of microorganisms in the compost during composting. Values are means of three replicates ± 1
Standard Error.

effect of the rapid increases in microbial activity in the Rhodococcus sp. and Mycobacterium sp. among others
first month manifested in higher decreases in concentra- that were not readily identified. The dominant fungal
tions in the second month. This delayed effect is species were Mucor, Rhizopus, Fusarium, Aspergillus,
attributed to the time used by the organisms to adapt to Penicillium and Pleurotus. By the end of the fourth month,
the hydrocarbon medium and also other factors such as the dominant species were mainly fungi with Pleurotus
high initial nitrogen content and high temperatures as sp., becoming more prominent. Fusarium, Aspergillus
discussed earlier. and Penicillium persisted in relatively low amounts at the
The rate of removal of hydrocarbons from the control end of incubation. Mucor and Rhizopus were not evident
experiment and the sewage sludge compost system in the compost at the end. Phanerochaete was isolated
showed a similar trend in the first month (Figures 6, 7 and after the fourth month. Three Pseudomonas species and
8). However, while removal continued very slowly for the Arthrobacter sp. were isolated after the fourth month.
rest of the experimental period in the control experiment, Although, there was a dynamic change in the microbial
it increased rapidly from the second month in the population of the compost system during the composting
compost system. period, Pleurotus, Phanerochaete, Fusarium, Pseudo-
Compared to earlier studies using a similar conta- monas and Arthrobacter were the most persistent genera
minated soil in which more than 20% of the 4- and 5- ring that were identified.
PAHs persisted at the end of eleven months of treatment
(Atagana, 2004a), all the tested components were
-1
removed below the remediation target (1 mg kg ) by the Conclusions
end of the incubation period.
Contrary to expectations, high nitrogen content and Although, sewage sludge accelerated the composting of
high temperatures had limited effects on the microbial garden refuse, their application in compost bioreme-
load and degradation capacity of the compost probably diation has not been fully explored. The results in the
due to the reasons discussed earlier (Figures 8). present experiment shows that under controlled conditions
At the start of composting, a mixed population of inoculation of compost containing hydrocarbon-conta-
bacteria and fungi dominated the compost. The dominant minated soil with sewage sludge can effectively accelerate
bacteria isolates were Pseudomonas sp., Bacillus sp., the removal of such contaminants from the soil matrix.
1522 Afr. J. Biotechnol.

140

120 Control experi ment

) 100
Concentration (mg kg-1)

-1
kg
g
(m 80
n
o
ti
a
tr 60
n
e
c
n
o
C 40

20

0
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19
Tim e (Months )

Phenol o-c resol m-c resol p-cresol

140

120 S ewage sl udge c om post

Concentration (mg kg-1)

) 100
1
-
g
k
g
( 80
m
n
o
it
ra
t 60
n
e
c
n
o 40
C

20

0
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19
Time (Months )

Phenol o-cresol m-cresol p-cresol

Figure 6. Changes in the concentrations of selected hydrocarbons (phenols) during composting. Values are means of
three replicates ± 1 Standard Error.
 Atagana 1523

300

2- and 3-ring P AHs i n control experiment

250

)
1
Concentration (mg kg-1)

-
g 200
k
g
(m
n
io
ta
150
tr
n
e
c
n 100
o
C

50

0
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19
Ti me (Mont hs)

Naphthale ne Anthracene Phenanthrene Pyrrole Fluorene

300

2- and 3-ring PAHs i n sewage sludge compost

250

)
-1 200
Concentration (mg kg-1)

g
k
g
m
(
n
o
it 150
a
rt
n
e
c
n 100
o
C

50

0
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19
Tim e (Month s)

Naphthalene Anthracene Phenant hrene Pyrrole Fluorene

Figure 7. Changes in the concentrations of selected hydrocarbons (2- and 3- ring PAHs) in the compost during
composting. Values are means of three replicates ± 1 Standard Error.

ACKNOWLEDGEMENTS suthu Technikon, Durban, South Africa; Department of

Microbiology and Plant Pathology, University of KwaZulu-
I wish to acknowledge the following departments for their Natal, Pietermaritzburg, South Africa; University of
support: Department of Analytical Chemistry, Mango- KwaZulu-Natal Experimental Farm, Ukulinga, Pieter-
1524 Afr. J. Biotechnol.

200

180 4- and 5- rin ged PA Hs in the con trol experim ent

160

) 140
Concentration (mg kg-1)

1
-
g
k
g 120
(m
n
o
ti 100
a
tr
n
e 80
c
n
o
C 60

40

20

0
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19
Ti me (M onth s)

Pyrene Chrysene Fluoranthene B enzo(a)pyrene

250

4- and 5-ringed PAHs in sewage sl udge compost

200

1 )
-
g
k
Concentration (mg kg-1)

g 150
m
(
n
o
ti
a
rt
n
e
c 100
n
o
C

50

0
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19
Ti me (M on ths)

Pyrene Chrysene Fluorant hene Benzo(a)pyrene

Figure 8. Changes in the concentrations of selected hydrocarbons (4- and 5- ring PAHs) in the compost
during composting. Values are means of three ± 1 Standard Error.

maritzburg, South Africa; and Department of Soil REFERENCES

Science, University of KwaZulu-Natal, Pietermaritzburg, Alef K (1995). Soil respiration. In Alef K, Nannipieri P (eds) Methods in
South Africa. Applied Soil Microbiology and Biochemistry. Academic Press,
 Atagana 1525

London. 214-219, Kubota H, Nakasaki K (1991). Accelerated thermophilic composting of

Alexander M (1999). Biodegradation and Bioremediation. Academic garbage. Biocycle 32: 66-68.
Press, San Diego. Lees ZM (1996). Bioremediation of oil-contaminated soil: A South
Anderson JG (1991). Treatment of wastes by composting, In Senior E African case study. Ph.D. Thesis, University of Natal, Pietermaritz-
(ed) Microbiology of Landfill sites. CRC Press Inc., Boca Raton. 59- burg, South Africa.
77. MacFaddin JF (1980). Biochemical tests for identification of medical
Atagana HI (2003). Bioremediation of creosote contaminated soil: a bacteria, Lippincot Williams and Wilkins, Baltimore.
pilot-scale landfarming evaluation. World J. Microbiol. Biotechnol. 19: Manios T, Maniadakis K, Kalogeraki M, Mari E, Stratakis E, Terzakis S,
571-581. Boytzakis P, Naziridis Y, Zampetakis L (2006). Efforts to explain and
Atagana HI (2004). Bioremediation of creosote-contaminated soil in control the prolonged thermophilic period in two-phase olive oil mill
South Africa by landfarming. J. Appl. Microbiol. 96: 510-520. sludge composting. Biodegradation 17: 285-292.
Atagana HI (2004). Co-composting of PAH-contaminated soil with Marin JA, Moremo JL, Hernandez T, Garcia C (2006). Bioremediation
poultry manure. Lett. Appl. Microbiol. 39: 163-168. by composting of heavy oil refinery sludge in semiarid conditions.
Atagana HI, Haynes RJ, Wallis FM (2003). Co-composting of soil Biodegradation. 17: 251-261.
heavily contaminated with creosote with cattle manure and mixed Piccinini S, Rossi L, Bonazzi G, Dall’Orso G (1996). The Emilia-
vegetable waste. Soil and Sediment Contamination: An Int. J. 12(6): Romaga Experiment in animal manure composting, In: de Bertoldi M,
889-899. Sequi P, Lemmes B, Papi T (eds) The Science of Composting.
Baker KH, Herson DS (1994). Bioremediation. McGraw-Hill, Toronto. Blackie Academic and Professional: London, pp. 1275-1280.
Bardos RP, Forsythe S, Westlake K (1996). The co-treatment of Potter CL, Glaser JA, Hermann R, Dosani MA (1999). Remediation of
municipal and industrial waste. In: de Bertoldi M, Sequi P, Lemmes contaminated East River sediment by composting technology. In:
B, Papi T (eds) The Science of Composting. Blackie Academic and Leeson A, Alleman BC (eds) Bioremediation Technologies for
Professional, London. 767-783. Polyclic Aromatic Hydrocarbon Compounds. The Fifth International
Barnett HL, Hunter BB (1972). Illustrated genera of imperfect fungi. In-situ and On-site Bioremediation Symposium. Battelle Press:
Burgess Publishing Company, Minneapolis. Columbus, pp. 31-36.
Combs SM, Peters JB, Zhang LS (2001). Micronutrient Status of Raper KB, Thom CA (1968). A manual of the penicillia. Hafner
Manure. in Wisconsin Forage Council Proceedings. University of Publishing Company: New York.
Wisconsin Press: Madison, pp. 42-50. Recommended Chemical Soil Test for the North Central Region (1998).
Diaz LF, Savage GM, Golueke CG (1996). Stabilization of hazardous North Central Regional Publication No. 221 (Revised), University of
wastes through biotreatment. In: de Bertoldi M Sequi P Lemmes B Missouri Agricultural Experiment Station, Columbia, Missouri.
Papi T (eds) The Science of Composting. Blackie Academic and Reid BJ, Jones KC, Semple KT, Fermor TR (1999). Bioremediation
Professional: London, pp. 1152-1156. potential of PAHs in compost. in Bioremediation technologies for
Eriksson M, Dalhammar G, Borg-Karlson AK (2000). Biological polyclic aromatic hydrocarbon compounds. The fifth international in
degradation of selected hydrocarbons in an old PAH/creosote situ and on-site bioremediation symposium, Leeson A, Alleman BC
contaminated soil from a gas work site. Appl. Microbiol. Biotechnol. eds., Battelle Press: Columbus, pp. 19-22.
53: 619-626. Schmitt M, Rhen G (2002). Fertilizing Cropland with Poultry Manure. in
Eweis JB, Ergas SJ, Chang DP, Schroeder ED (1999). Biodegradacion Regents of The University of Minnesota. FO-5881 C.
de compuestos concretos principios de biorrecuparacion. McGraw Sutherland JB, Rafti F, Khan AA, Cerniglia CE (1995). Mechanisms of
Hill, Spain, pp. 131-147. polycyclic aromatic hydrocarbon degradation. In: Young LY, Cerniglia
Fan C, Tafuri AN (1994). Engineering applications of biooxidation CE (eds) Microbial transformation and degradation of toxic organic
processes for treating petroleum contaminated soil. In: Wise DL, chemicals. Wiley Liss, New York, pp. 269-306,
Trantolo DJ (eds) Remediation of Hazardous Waste Contaminated US EPA (1996). Total recoverable petroleum hydrocarbons by infrared
Soils, Marcel Dekker Inc, New York, p. 929. spectrophotometry. EPA method No. 8440.
Feinstein MS, Miller FC, Strom PE (1986). Waste treatment composting Valo R, Salkinoja-Salonen M (1986). Bioremediation of chlorophenol-
as a controlled system, In: Rhelm HJ, Reed G (eds) Biotechnology contaminated land. Appl. Microbiol. Biotechnol. 53: 619-626.
Vol. 8 Microbial degradation. VCH Publishers, New York, pp. 363- Wilson GB, Sikora LJ, Parr JF (1983). Composting of chemical
369. industrial wastes prior to land application, In: Parr JF, Marsh PB, Kla,
Forster JC (1995a). Determination of the gravimetric water content and JM (eds) Treatment of hazardous wastes. Noyes Data Corporation,
soil dry mass. In Alef K, Nannipieri P (eds) Methods in Applied Soil New Jersey, pp. 263-273.
Microbiology and Biochemistry. Academic Press, London, pp. 105-
106.
Forster JC (1995b). Determination of total organic carbon. In Alef K,
Nannipieri P (eds) Methods in Applied Soil Microbiology and
Biochemistry. Academic Press, London, pp. 59-60.
Forster JC (1995c). Determination of total nitrogen. In Alef K, Nannipieri
P (eds). Meth. Appl. Soil Microbiol. Biochem. Academic Press,
London, pp. 79-80.
Garcia-Gomez A, Roig A, Bernal MP (2003). Composting of the solid
fraction of olive oil mill wastewater with olive leaves: organic matter
degradation and biological activity. Bioresour. Technol. 86: 59-64,
Hill DW, McCarty PL (1967) Anaerobic degradation of selected
chlorinated pesticides. J. Water Poll. Cont. Fed. 39: 1259-1277.
Holt JG (1994). Bergey’s Manual of Determinative Bacteriology.
Lippincott, Williams and Wilkins: Baltimore.