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IOSR Journal of Engineering (IOSRJEN) www.iosrjen.org
ISSN (e): 2250-3021, ISSN (p): 2278-8719
Vol. 05, Issue 01 (January 2015), ||V2|| PP 07-15
International organization of Scientific Research 7 | P a g e
Microbial Assessment of Air In The Vicinity Of Some Dump Sites
in Delta State
Jude C. Igborgbor1
*, Gideon I. Ogu2
1
Department of Biology, College of Education Agbor, Delta State, Nigeria
2
Department of Biological Sciences, Novena University, Ogume, Delta State, Nigeria
Abstract: - The microbial loads of air in the vicinity of various dumpsites in Delta State, Nigeria, were
conducted for a period of 6 months (July - December, 2012) using standard pour plate and spread plate
microbiological techniques. Results showed that all the tested parameters decreased with distance away from the
dumpsites and the microbial loads were higher than regulatory limits. The heterotrophic bacteria count ranged
from 1.41 x 108
to 29.2 x 1014
cfu/ml, while fungal counts ranged from 1.12 x 108
to 1.39 x 1014
cfu/ml.
Bacterial counts were higher in the months of July, August and September, while fungi were more in the months
of October to December. The bacterial genera isolated were Staphylococcus, Streptococcus, Micrococcus,
Bacillus, Pseudomonas, Escherichia coli, and Klebsiella species, while the fungal isolates were Penicillium,
Aspergillus, Alternaria, Mucor, Curvularia, Rhizopus and Cladosporium sp. Statistical analysis showed that
there were significant differences (P<0.05) between the microbial load of the various dumpsites, periods and
distance of sampling sites. This study showed that potential airborne pathogens not only abound in the vicinity
of waste dumpsites but also decreased with increasing distance from the dump sites. It is therefore
recommended that dumpsites be located at a minimum 1km distance away from residential quarters.
Keywords: - Microbial loads, dumpsites, heterotrophic bacteria count, residential quarters
I. INTRODUCTION
Indiscriminate dumping of waste has become a common practice in Nigeria and other African
countries. In fact, virtually every available space close to residential areas, roads, farms, markets, hospital and
churches in many Nigerian communities now serve as waste dump sites [1]. The lack of proper refuse disposal
and waste treatment facilities in these areas, therefore, makes the populace to be at risk of infections from
pathogens that are airborne in the dumpsite areas.The World Health Organization estimates that about two
million people die prematurely every yearas a result of air pollution, while many more suffer from breathing
ailments, heart disease, lung infections and even cancer [2].
Over the years, studies have been extensively carried out on the microbial qualities of air in hospital
environments [3-11]. In order to develop appropriate air quality management plans, however, it is necessary first
to have reliable information about the state of airborne bacteria and fungi especially in the vicinity of waste
dumpsites. Therefore, this study was undertaken to determine the microbial load of air in the vicinity of some
waste dumpsites in selected towns (Warri, Agbarho and Agbor,) in Delta State, Nigeria.
II. MATERIALS AND METHODS
2.1 Study Area
The study area is located within Nigeria Delta in the Southern portion of Nigeria (Fig. 1). Warri is between
latitude 50
32’N and 50
40’ N and longitude 50
42’E and 5’50’E, while Agbor is situated between latitude 6.00’N
and longitude 60
05’ E and 60
25’N and longitude 60
05’E and 60
25’ E. Agbarho is between latitude 50
34’N and
50
40’ N and longitude 542’E and 5’50’E. All the communities are in Delta State, Nigeria.
Microbial Assessment of Air In The Vicinity Of Some Dump Sites in Delta State
International organization of Scientific Research 8 | P a g e
Fig. 1: Map of Delta State Showing Warri, Agbarho and Agbor.
2.2 Sample Collection
Air samples were collected monthly for period of 6 months (July – December) from the various
dumpsites in Agbor, Warri and Agbarho. At each sampling area, wide mouth bottles containing sterile peptone
water was exposed for 15 minutes in the dumpsite and at distances of 100m, 200m, 300m, 400m and 500m
away from the dumpsite. In all ninety air samples were collected during the period and the sample bottles were
corked and taken to the laboratory for analysis.
2.3 Culturing and Enumeration of Bacteria in Air Samples
Each sterile peptone water exposed to air was thoroughly shaken to obtain a homogenous sample. An
aliquot (1.0 ml) was diluted serially to 10-15
dilution [12]. From the dilution of 10-8
, 10-12
and 10-14
of each
sample, 0.1 ml aliquot was transferred aseptically onto freshly prepared Nutrient agar plates. The inoculated
plates were inverted and incubated at 37 o
C for 24 hrs after which the plates were examined for growth. The
discrete colonies which developed were counted and the average counts for duplicate cultures were recorded as
aerobic heterotrophic bacteria in the sample.
2.4 Isolation, characterization and identification of bacteria in the air samples
Pure cultures of bacteria were obtained by aseptically streaking representative colonies of different
morphological types, which appeared on the cultured plates onto freshly prepared Nutrient agar plates and
MacConkey agar plates and incubated at 37 o
C for 24 hrs. Discrete bacteria colonies which developed were sub-
cultured onto Nutrient agar slopes and incubated at 28 o
C for 24 hrs. These served as pure stock cultures for
subsequent characterization and identification via physiological and biochemical tests [13].
2.5 Culturing and enumeration of fungi in air samples
Each air sample was shaken thoroughly to obtain a homogenous sample. Aliquot (1.0 ml) of it was
diluted serially to 10-15
dilution. From the dilution of 10–8
, 10-12
and 10-14
of each air sample, 0.1 ml aliquot was
transferred aseptically onto freshly prepared Potato dextrose agar plates containing 0.2 ml of 0.5 % Ampicillin
to inhibit the growth of bacteria [14]. The inoculum was spread with a sterile bent glass rod. The inoculated
plates were inverted and incubated at (room temperature) 28 ± 2 0
C for 5 days. The colonies which developed
were counted and the average counts for duplicate cultures were recorded as viable fungal counts in the sample.
Microbial Assessment of Air In The Vicinity Of Some Dump Sites in Delta State
International organization of Scientific Research 9 | P a g e
2.6 Isolation, characterization and identification of fungi in air samples.
Pure cultures of fungi were obtained by subculturing discrete colonies onto freshly prepared Potato
Dextrose Agar plates and inoculated at room temperature (28± 2 0
C) for 5 days. The fungal isolates which
developed were further sub cultured onto agar slopes and incubated at room temperature. The isolates which
developed were pure cultures which were stored in the refrigerator (4 o
C) as stock cultures for subsequent
characterization via macroscopic and microscopic examination. The identification of fungal isolates was done
by comparing the result of their cultural and morphological characteristics with those of known taxa [14].
2.7 Statistical Analysis
Data obtained was analysed using Microsoft 2003 excel programme. Analysis of variance ANOVA (F-
test) was done to determine the statistical difference in microbial count between the sampling stations while
seasonal variation in bacterial and fungal counts for the air samples at 95 % confidence level was determined
using student’s t - test.
III. RESULTS
3.1 Bacterial Load for Air Samples
The bacterial load for the air samples from the dumpsites in Warri, Agbarho and Agbor are presented in
Figs 4.1, 4.2 and 4.3 respectively. It was observed that the bacterial counts decreased with distance from the
dumpsite and were higher in the months of July, August and September, but decreased slightly in the months of
October to December. The bacterial counts in the air samples from the dumpsite in July - September ranged
from 15.47 to 14.22 log10 cfu/ml while the bacterial counts in the air samples from the dumpsite in October-
December ranged from 14.37 to 12.37 log10 cfu/ml. Similarly the bacterial counts in the air samples taken from
the various distances ranged from 13.58 to 8.44 log10cfu/ml in July - September while the value ranged from
14.37 to 7.44 log10 cfu/ml in October-December. Statistical analysis showed that there were significant
differences (P<0.05) between the bacteria load of the various sampling stations and seasonal variations
Fig 2: Bacterial Counts for Warri Air Samples
Key: DS = dumpsite, ST1 = station 1 (100m from dumpsite), ST2 = station 2 (200m away from dumpsite), ST3
= station 3 (300m away from dumpsite), ST4 = station 4 (400m away from dumpsite), ST5 = station 5 (500m
away from dumpsite).
Fig 3: Bacterial Counts for Agbarho Air Samples.
Microbial Assessment of Air In The Vicinity Of Some Dump Sites in Delta State
International organization of Scientific Research 10 | P a g e
Key: DS = dumpsite, ST1 = station 1 (100m from dumpsite), ST2 = station 2 (200m away from dumpsite), ST3
= station 3 (300m away from dumpsite), ST4 = station 4 (400m away from dumpsite), ST5 = station 5 (500m
away from dumpsite).
Fig. 4: Bacterial counts for Agbor Air Samples.
Key: DS = dumpsite, ST1 = station 1 (100m from dumpsite), ST2 = station 2 (200m away from dumpsite), ST3
= station 3 (300m away from dumpsite), ST4 = station 4 (400m away from dumpsite), ST5 = station 5 (500m
away from dumpsite).
3.2 Fungal Count for Air Samples
The fungal counts of the air samples from the dumpsites in Warri, Agbarho and Agbor are presented in
Figs 5, 6 and 7. It was observed that the fungal counts decreased with distance from the dumpsite and the fungal
counts increased in the months of November and December. The fugal counts in the ambient air of the dumpsite
in July - September ranged from 12.58 to 10.58 log10 cfu/ml and the fungal load in the air samples from the
dumpsite in October-December ranged from 13.67 to 12.57 log10 cfu/ml. Similarly the total fungal counts in the
air samples taken from the various distances ranged from 11.78 to 8.04 log10 cfu/ml in July - September while
the value ranged from 13.49 to 8.48 log10 cfu/ml in October-December. Statistical analysis showed that there
were significant differences (P<0.05) between the fungal loads of the various sampling stations and during the
different seasons.
Fig 5: Fungal Counts for Warri Air Samples
Key: DS = dumpsite, ST1 = station 1 (100m from dumpsite), ST2 = station 2 (200m away from dumpsite), ST3
= station 3 (300m away from dumpsite), ST4 = station 4 (400m away from dumpsite), ST5 = station 5 (500m
away from dumpsite).
Fig 6: Fungal Counts for Agbarho Air Samples
Microbial Assessment of Air In The Vicinity Of Some Dump Sites in Delta State
International organization of Scientific Research 11 | P a g e
Key: DS = dumpsite, ST1 = station 1 (100m from dumpsite), ST2 = station 2 (200m away from dumpsite), ST3
= station 3 (300m away from dumpsite), ST4 = station 4 (400m away from dumpsite), ST5 = station 5 (500m
away from dumpsite).
Fig 7: Fungal Counts for Agbor Air Sample
Key: DS = dumpsite, ST1 = station 1 (100m from dumpsite), ST2 = station 2 (200m away from dumpsite), ST3
= station 3 (300m away from dumpsite), ST4 = station 4 (400m away from dumpsite), ST5 = station 5 (500m
away from dumpsite).
3.3 Bacterial Isolates from Air Samples
The bacterial genera isolated from the air sample were Staphylococcus sp, Streptococcus sp, Micrococcus sp,
Bacillus sp, Pseudomonas sp, Escherichia coli, and Klebsiella sp were isolated from the air samples (Table 1)
Table 1:Identification and characterization of bacteria from air samples
Isolate A B C D E F G
Cultural
characteristics
Creamy
converse
colonies with
smooth edges
Cocci clusters Round clusters
dented
white flat
& smooth
colonies
Dented and
raised
colonies
Pink
convex
Smooth
colonies
Flat
yellow
swarming
colonies
Pigmentation Cream Cream Yellow Cream Green Pink cream
Cellular
morphology
Cocci Cocci Cocci Rods Rods Rods Rods
Gram Stain + + + + _ _ _
Aerobic growth + + + + + + +
Anaerobic growth + + _ _ _ _ +
Motility Test _ _ _ + + + +
CatalaseTest + _ + + + + +
OxidaseTest _ _ + _ + _ _
Urease Test + _ + _ + _ +
Endospore Test _ _ _ + _ _ _
IndoleTest + _ _ _ _ + _
Microbial Assessment of Air In The Vicinity Of Some Dump Sites in Delta State
International organization of Scientific Research 12 | P a g e
Key: + =positive, – =negative
3.4 Distribution of Bacterial Isolates in Air Samples
The distribution of the bacterial isolates obtained from the air samples in wet and dry seasons are presented in
Tables 2 and 3 respectively. It was observed that Streptococcus sp, Escherichia coli, Staphylococcus sp,
Pseudomonas sp, Micrococcus sp, Klebsiella sp and Bacillus sp were more abundant in the air in the dumpsite
and decreased with distanceaway from the dumpsites in the wet season while Micrococcus sp and Bacillus sp
were abundant in both wet and dry seasons.
Table 2:Distribution of air bacterial isolates in wet season
Organism Dumpsite ST1 ST 2 ST3 ST 4 ST 5
Streptococcus sp +++ +++ ++ ++ ++ ++
Bacillus sp +++ +++ ++ ++ ++ ++
Escherichia coli +++ +++ ++ ++ ++ ++
Micrococcus sp +++ +++ ++ ++ ++ ++
Staphylococcus sp +++ +++ ++ ++ ++ ++
Pseudomonas sp. +++ +++ ++ ++ ++ ++
Klebsiella sp +++ +++ ++ ++ ++ ++
Key: – Absent, + Rare, ++ Intermediate, +++ large in number
Table 3:Distribution of air bacterial isolates in dry season
Key: – Absent, + Rare, ++Intermediate, +++large in number
3.5 Fungal Isolates from Air Samples
The fungal isolates from the air, samples are presented in Table 4. The fungal genera isolated were Alternaria
sp, Aspergillus sp, Cladosporium sp, Curvularia sp, Mucor sp, Penicillium sp and Rhizopus sp.
Citrate Test + + + + + + _
Glucose
Fermentation test
+ _ + + + + +
Lactose
Fermentation test
_ _ _ _ _ + +
H2S
production test
_ _ _ _ _ _ _
Organism
identified
Staphylococcus
sp
Streptococcus
sp
Micrococcus
sp
Bacillus
sp
Pseudomo
nas sp
E. coli. Klebsiella
sp
Organism Dumpsite ST1 ST 2 ST3 ST 4 ST 5
Streptococcus sp
++ ++ ++ ++ + +
Bacillus sp. +++ +++ +++ +++ ++ ++
Escherichia coli
++ ++ ++ ++ + +
Micrococcus sp +++ +++ +++ +++ ++ +
Staphylococcus sp ++ ++ ++ ++ + +
Pseudomonas sp. ++ ++ ++ + + +
Klebsiella sp ++ ++ ++ + + -
Microbial Assessment of Air In The Vicinity Of Some Dump Sites in Delta State
International organization of Scientific Research 13 | P a g e
3.6 Distribution of Fungal Isolates in Air Samples
The distribution of the fungal isolates obtained from the air samples in wet and dry seasons are presented in
Tables 5 and 6 respectively. It was observed that all the fungal isolates were abundant in the dry season while
Aspergillus sp and Mucor sp were abundant in both wet and dry seasons.
Table 5:Distribution of air fungal isolates in wet season
Organism Dumpsite ST1 ST2 ST3 ST4 ST 5
Aspergillus sp +++ ++ ++ ++ ++ ++
Cladosporium sp ++ + + + + +
Curvularia sp ++ + + + + +
Mucor sp +++ ++ ++ ++ ++ ++
Rhizopus sp ++ + + + + +
Penicillium sp ++ + + + + _
Alternaria sp ++ + + + + +
Key: – Absent, + Rare, ++Intermediate, +++large in number
Table 6:Distribution of air fungal isolates in dry season
Organism Dumpsite ST1 ST2 ST3 ST4 ST 5
Aspergillus sp +++ ++ ++ ++ ++ ++
Cladosporium sp +++ ++ ++ ++ ++ ++
Curvularia sp +++ ++ ++ ++ ++ ++
Microbial Assessment of Air In The Vicinity Of Some Dump Sites in Delta State
International organization of Scientific Research 14 | P a g e
Mucor sp +++ ++ ++ ++ ++ ++
Rhizopus sp +++ ++ ++ ++ ++ ++
Penicillium sp +++ ++ ++ ++ ++ ++
Alternaria sp +++ ++ ++ ++ ++ ++
Key: – Absent, + Rare, ++Intermediate, +++large in number
IV. DISCUSSION
The microbial loads of the air samples taken from the dumpsites were higher than the normal
atmospheric concentration of the microorganisms as the reported average level of the microbes in the ambient
air is 3.0 log10cfu/ml [15]. This is an indication of the extent of microbial pollution of waste dump sites in the
study sites
The microorganisms isolated from the air samples taken from the various locations (Warri, Agbarho and
Agbor), and distances sampled were similar in composition and distribution with those isolated from the air
samples taken from the dumpsite. Generally the microbial load decreased with distances away from the
dumpsite. The bacterial load for the air samples from Warri, Agbarho and Agbor presented in Figs 2- 4 showed
that the bacteria counts decreased with distance from the dumpsite. It was equally observed that the bacterial
counts were higher in the months of July August and September (wet season) and decreased slightly in the
months of October, November and December (dry season). The decreasing bacterial counts with distance away
from the dumpsite could be due to increased microbial activity in the dumpsite as a result of putrefaction and
increased decomposition of organic matter in the vicinity of the dumpsite. It could also be as a result of
household and industrial wastes deposited in the waste dumpsites as well as contaminants generated naturally
that were propelled through the air, such as particles of dust and soil microbial spores in the air within the
dumpsites. These results agree with the report of McCarthy [16], who listed these amongst others as possible
sources of air contaminants.
The observed increased trend in the bacterial counts in wet season could be as a result of increased rainfall in the
months of July, August and September and decreased rainfall in the months of October, November and
December leading to reduced water activity. These results agree with the reports of Obire et al. [17], who stated
that seasonal variations favour physiological types.
The fungal load for the air samples from Warri, Agbarho and Agbor as presented in Figs 5-7 showed that the
fungal counts decreased with distance away from the dumpsites. It was equally observed that the fungal counts
were lower in the months of July August and September (wet season) and increased slightly in the months of
October, November and December (dry season). The decreasing fungal count with distance away from the
dumpsite could be due to same reasons propounded for bacterial counts above. These results agreed with the
report of McCarthy [16], who reported similar suggestions
The observed trend in the fungal counts could be due to spore formation due to increased rainfall in the months
of July, August and September and decreased rainfall in the months of October, November and December.
These findings also agree with the reports of earlier researchers [17] who opined that seasonal variations favour
physiological types.
The bacterial and fungal genera encountered in this study had been reported by previous workers [18-
20], as the possible microbial isolates from the air. The isolated bacteria were more abundant in wet season than
in the dry season. This could possibly be due to the atmospheric particles to which the microbes are attached
being deposited by the process of rainfall. The increased water activity therein provides favourable conditions
for bacteria to thrive and multiply. Bacillus sp and Micrococcus sp were more abundant in the air sample both in
wet and dry seasons. The presence and prevalence of some of these species of bacteria in the dumpsite could be
as a result of the presence of damp organic materials, materials impregnated with water, food and food products
and spores of microorganisms propelled through the air. These results agree with the report of Osha [21] and
Sola [22], who reported these as possible sources of air microflora. These bacteria, according to Douwes [23]
can cause different forms of bacterial pneumonia, influenza and gastrointestinal diseases.
The general trend of the isolated fungi showed that they were more abundant in dry season than in the wet
season. This could be as a result of various climatic and local topographic factors including reduced rainfall
leading to reduced water activity which favour germination of spores.It could also be as a result of the wind
action and harsh weather conditions experienced during the dry season. Cladosporium sp, Penicillium sp, Mucor
sp, Alternaria sp, Aspergillus sp, Rhizopus sp and Curvularia sp were observed to be predominant in the air
samples during the dry season while Aspergillus sp and Mucor sp species were predominant in both the wet and
Microbial Assessment of Air In The Vicinity Of Some Dump Sites in Delta State
International organization of Scientific Research 15 | P a g e
dry season. These results agree with the report of Prescott et al. [24], who listed these amongst the most
common allergenic moulds associated with man and live stocks. Obire et al. [17] also identified Aspergillus sp
as one of the most common fungi.
The present study has revealed that the dumpsite had impact on both the microbial load and quality of
such environment. Also, the microbial load of air in dumpsites decreases with increase in distance from the
dump sites. Airborne bacteria at dumpsites are more prevalent in wet seasons than in the dry seasons, the reverse
being the case for airborne fungi. It is therefore recommended that dumpsites should be located at about I km
away from residential areas to avoid contamination of the air. Additionally, educational programmes should be
organized for public awareness of impact of indiscriminate dumping of refuse.
REFERENCES
[1] F. C. Akharayi and F.O. Omoya. Microflora of Refuse Dumps and sites in five localities in Ondo state, Nigeria. Inter.
J. Environ. 2(1-2), 2004, 27-32.
[2] R. Madhukar and S. Srikantaswamy. Assessment of Air Quality in Bidadi Industrial Area, Ramanagaram District,
Karnataka, India. J. Environ. Sci., Comp. Sci. Eng. Technol. 2(4), 2013, 1135-1140.
[3] A. A. Jaffal, H. Nsanze, A. Benar, A. S. Ameen, I. M. Banat and A. A. EL-Moghett. Hospital airborne microbial
pollution in a desert country. Environ. Inter., 1997, 23(2):67-172.
[4] T. T. Adebolu and K. J. Vhriterhire. Survey of the microbial flora of the Ondo State Specialist Hospital Environment,
Akure, Nigeria. Nig. J. Microbiol., 16(112), 2002, 91-94.
[5] J. Dutkiewicz and M. Augustowska. Variability of airborne microflora in a hospital ward with a period of one year.
Ann. Agric. Environ. Med., 13, 2006, 99-106.
[6] D. H. Tambeker, P. B. Gulhane and D. D. Bhakare. Studies on environmental monitoring of microbial air flora in
hospitals. J. Med. Sci. 7(1), 2007, 67-72.
[7] F. O. Ekhaise, O. U. Ighosewe and O. D. Ajakpori. Hospital indoor airborne microflora in private and government
owned hospitals in Benin City, Nigeria. World. J. Med. Sci., 3(1), 2008, 34-38.
[8] S. Sudharsan, P. Srikath, M. Sheela and R. Stienberg. Study of indoor air quality in hospitals in South Chennai, India.
Ind. Blt. Environ. 17, 2008, 435-441
[9] F. O. Ekhaise, E. E. Isitor, O. Idehen, and A. O. Emoghene. Airborne Microflora in the Atmosphere of an Hospital
Environment of University of Benin Teaching Hospital (UBTH), Benin City, Nigeria. World J. Agri. Sci., 6(2), 2010,
166-170
[10] S. A. Awosika, F. A. Olajunbu, and N. A. Amusa. Microbiological assessment of indoor air of a teachinghospital in
Nigeria. Asian Pac. J. Trop. Bio. 16, 2012, 465-468.
[11] M. N. O. Omoigberale, O. O. Amengialue and M. I. Iyamu. Microbiological assessment of Hospital Indoor Air
Quality in Ekpoma, Edo State, Nigeria. Global Res. J. Microbiol., 4(1), 2014, 1-5.
[12] M. Cheesebrough. District Laboratory Practice in Tropical Countries. Part 2, 2000, 32-33.
[13] M. Cheesbrough. District Laboratory Practice In Tropical Countries, part 2, second edition Cambridge University
press, The Edinburgh Building, Cambridge, United Kingdom, 2006, 38, 62-69.
[14] W. F. Harrigan, and M. E. McCance. Laboratory Methods in Food and Dairy Microbiology. 8th
Edition Academic
press. London.1990, 210
[15] Y. Panthi, and U. Shrestha. Microbial Air Pollution A case study of sodisol sanitary landfill site. Asian J. Appl. Sci.
1(1), 2008, 94-97.
[16] J. McCarthy. Indoor Air Quality Handbook. Mc-Graw Hill London. 2001, 290.
[17] O. Obire. A. O. Nwaubeta and S. B. N. Adue. Microbial Community of a Waste Dumpsite. J. Appl. Sci. Environ.
Manag. 2, 2002, 73-82.
[18] D. K. G. Ayanru. Global Impact Air Spore. Appl. Microbiol. 6, 1981, 107-117.
[19] K. J. Ryan, C. G. Ray and S. Sherris. Medical Microbiology. 4th
edition: McGraw Hill, New York. 2004, 623-625.
[20] P. M. Kirk, P. F. Cannon, D. W. Minter and J. A. Stalpers. Dictionary of the Fungi .10th
ed. Wallingford: 2008, 222.
[21] S. Osha. Microbial contamination of the Environment. J. Curr. Sci. 89, 1999, 101-109
[22] G. Sola. Indoor Air Quality. J. Appl. Environ. Microbiol. 65, 2000, 423-432.
[23] J. Douwes. Bioaerosol health effects and exposure assessment: Progress and prospects. Ann. Occup. Hyg., 47(3),
2003, 187-200.
[24] L. M. Prescott, J. P. Harley, and A. D. Klein. Microbiology. 6th Edition. McGraw-Hill, New York. 2005, 962 - 965

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  • 1. IOSR Journal of Engineering (IOSRJEN) www.iosrjen.org ISSN (e): 2250-3021, ISSN (p): 2278-8719 Vol. 05, Issue 01 (January 2015), ||V2|| PP 07-15 International organization of Scientific Research 7 | P a g e Microbial Assessment of Air In The Vicinity Of Some Dump Sites in Delta State Jude C. Igborgbor1 *, Gideon I. Ogu2 1 Department of Biology, College of Education Agbor, Delta State, Nigeria 2 Department of Biological Sciences, Novena University, Ogume, Delta State, Nigeria Abstract: - The microbial loads of air in the vicinity of various dumpsites in Delta State, Nigeria, were conducted for a period of 6 months (July - December, 2012) using standard pour plate and spread plate microbiological techniques. Results showed that all the tested parameters decreased with distance away from the dumpsites and the microbial loads were higher than regulatory limits. The heterotrophic bacteria count ranged from 1.41 x 108 to 29.2 x 1014 cfu/ml, while fungal counts ranged from 1.12 x 108 to 1.39 x 1014 cfu/ml. Bacterial counts were higher in the months of July, August and September, while fungi were more in the months of October to December. The bacterial genera isolated were Staphylococcus, Streptococcus, Micrococcus, Bacillus, Pseudomonas, Escherichia coli, and Klebsiella species, while the fungal isolates were Penicillium, Aspergillus, Alternaria, Mucor, Curvularia, Rhizopus and Cladosporium sp. Statistical analysis showed that there were significant differences (P<0.05) between the microbial load of the various dumpsites, periods and distance of sampling sites. This study showed that potential airborne pathogens not only abound in the vicinity of waste dumpsites but also decreased with increasing distance from the dump sites. It is therefore recommended that dumpsites be located at a minimum 1km distance away from residential quarters. Keywords: - Microbial loads, dumpsites, heterotrophic bacteria count, residential quarters I. INTRODUCTION Indiscriminate dumping of waste has become a common practice in Nigeria and other African countries. In fact, virtually every available space close to residential areas, roads, farms, markets, hospital and churches in many Nigerian communities now serve as waste dump sites [1]. The lack of proper refuse disposal and waste treatment facilities in these areas, therefore, makes the populace to be at risk of infections from pathogens that are airborne in the dumpsite areas.The World Health Organization estimates that about two million people die prematurely every yearas a result of air pollution, while many more suffer from breathing ailments, heart disease, lung infections and even cancer [2]. Over the years, studies have been extensively carried out on the microbial qualities of air in hospital environments [3-11]. In order to develop appropriate air quality management plans, however, it is necessary first to have reliable information about the state of airborne bacteria and fungi especially in the vicinity of waste dumpsites. Therefore, this study was undertaken to determine the microbial load of air in the vicinity of some waste dumpsites in selected towns (Warri, Agbarho and Agbor,) in Delta State, Nigeria. II. MATERIALS AND METHODS 2.1 Study Area The study area is located within Nigeria Delta in the Southern portion of Nigeria (Fig. 1). Warri is between latitude 50 32’N and 50 40’ N and longitude 50 42’E and 5’50’E, while Agbor is situated between latitude 6.00’N and longitude 60 05’ E and 60 25’N and longitude 60 05’E and 60 25’ E. Agbarho is between latitude 50 34’N and 50 40’ N and longitude 542’E and 5’50’E. All the communities are in Delta State, Nigeria.
  • 2. Microbial Assessment of Air In The Vicinity Of Some Dump Sites in Delta State International organization of Scientific Research 8 | P a g e Fig. 1: Map of Delta State Showing Warri, Agbarho and Agbor. 2.2 Sample Collection Air samples were collected monthly for period of 6 months (July – December) from the various dumpsites in Agbor, Warri and Agbarho. At each sampling area, wide mouth bottles containing sterile peptone water was exposed for 15 minutes in the dumpsite and at distances of 100m, 200m, 300m, 400m and 500m away from the dumpsite. In all ninety air samples were collected during the period and the sample bottles were corked and taken to the laboratory for analysis. 2.3 Culturing and Enumeration of Bacteria in Air Samples Each sterile peptone water exposed to air was thoroughly shaken to obtain a homogenous sample. An aliquot (1.0 ml) was diluted serially to 10-15 dilution [12]. From the dilution of 10-8 , 10-12 and 10-14 of each sample, 0.1 ml aliquot was transferred aseptically onto freshly prepared Nutrient agar plates. The inoculated plates were inverted and incubated at 37 o C for 24 hrs after which the plates were examined for growth. The discrete colonies which developed were counted and the average counts for duplicate cultures were recorded as aerobic heterotrophic bacteria in the sample. 2.4 Isolation, characterization and identification of bacteria in the air samples Pure cultures of bacteria were obtained by aseptically streaking representative colonies of different morphological types, which appeared on the cultured plates onto freshly prepared Nutrient agar plates and MacConkey agar plates and incubated at 37 o C for 24 hrs. Discrete bacteria colonies which developed were sub- cultured onto Nutrient agar slopes and incubated at 28 o C for 24 hrs. These served as pure stock cultures for subsequent characterization and identification via physiological and biochemical tests [13]. 2.5 Culturing and enumeration of fungi in air samples Each air sample was shaken thoroughly to obtain a homogenous sample. Aliquot (1.0 ml) of it was diluted serially to 10-15 dilution. From the dilution of 10–8 , 10-12 and 10-14 of each air sample, 0.1 ml aliquot was transferred aseptically onto freshly prepared Potato dextrose agar plates containing 0.2 ml of 0.5 % Ampicillin to inhibit the growth of bacteria [14]. The inoculum was spread with a sterile bent glass rod. The inoculated plates were inverted and incubated at (room temperature) 28 ± 2 0 C for 5 days. The colonies which developed were counted and the average counts for duplicate cultures were recorded as viable fungal counts in the sample.
  • 3. Microbial Assessment of Air In The Vicinity Of Some Dump Sites in Delta State International organization of Scientific Research 9 | P a g e 2.6 Isolation, characterization and identification of fungi in air samples. Pure cultures of fungi were obtained by subculturing discrete colonies onto freshly prepared Potato Dextrose Agar plates and inoculated at room temperature (28± 2 0 C) for 5 days. The fungal isolates which developed were further sub cultured onto agar slopes and incubated at room temperature. The isolates which developed were pure cultures which were stored in the refrigerator (4 o C) as stock cultures for subsequent characterization via macroscopic and microscopic examination. The identification of fungal isolates was done by comparing the result of their cultural and morphological characteristics with those of known taxa [14]. 2.7 Statistical Analysis Data obtained was analysed using Microsoft 2003 excel programme. Analysis of variance ANOVA (F- test) was done to determine the statistical difference in microbial count between the sampling stations while seasonal variation in bacterial and fungal counts for the air samples at 95 % confidence level was determined using student’s t - test. III. RESULTS 3.1 Bacterial Load for Air Samples The bacterial load for the air samples from the dumpsites in Warri, Agbarho and Agbor are presented in Figs 4.1, 4.2 and 4.3 respectively. It was observed that the bacterial counts decreased with distance from the dumpsite and were higher in the months of July, August and September, but decreased slightly in the months of October to December. The bacterial counts in the air samples from the dumpsite in July - September ranged from 15.47 to 14.22 log10 cfu/ml while the bacterial counts in the air samples from the dumpsite in October- December ranged from 14.37 to 12.37 log10 cfu/ml. Similarly the bacterial counts in the air samples taken from the various distances ranged from 13.58 to 8.44 log10cfu/ml in July - September while the value ranged from 14.37 to 7.44 log10 cfu/ml in October-December. Statistical analysis showed that there were significant differences (P<0.05) between the bacteria load of the various sampling stations and seasonal variations Fig 2: Bacterial Counts for Warri Air Samples Key: DS = dumpsite, ST1 = station 1 (100m from dumpsite), ST2 = station 2 (200m away from dumpsite), ST3 = station 3 (300m away from dumpsite), ST4 = station 4 (400m away from dumpsite), ST5 = station 5 (500m away from dumpsite). Fig 3: Bacterial Counts for Agbarho Air Samples.
  • 4. Microbial Assessment of Air In The Vicinity Of Some Dump Sites in Delta State International organization of Scientific Research 10 | P a g e Key: DS = dumpsite, ST1 = station 1 (100m from dumpsite), ST2 = station 2 (200m away from dumpsite), ST3 = station 3 (300m away from dumpsite), ST4 = station 4 (400m away from dumpsite), ST5 = station 5 (500m away from dumpsite). Fig. 4: Bacterial counts for Agbor Air Samples. Key: DS = dumpsite, ST1 = station 1 (100m from dumpsite), ST2 = station 2 (200m away from dumpsite), ST3 = station 3 (300m away from dumpsite), ST4 = station 4 (400m away from dumpsite), ST5 = station 5 (500m away from dumpsite). 3.2 Fungal Count for Air Samples The fungal counts of the air samples from the dumpsites in Warri, Agbarho and Agbor are presented in Figs 5, 6 and 7. It was observed that the fungal counts decreased with distance from the dumpsite and the fungal counts increased in the months of November and December. The fugal counts in the ambient air of the dumpsite in July - September ranged from 12.58 to 10.58 log10 cfu/ml and the fungal load in the air samples from the dumpsite in October-December ranged from 13.67 to 12.57 log10 cfu/ml. Similarly the total fungal counts in the air samples taken from the various distances ranged from 11.78 to 8.04 log10 cfu/ml in July - September while the value ranged from 13.49 to 8.48 log10 cfu/ml in October-December. Statistical analysis showed that there were significant differences (P<0.05) between the fungal loads of the various sampling stations and during the different seasons. Fig 5: Fungal Counts for Warri Air Samples Key: DS = dumpsite, ST1 = station 1 (100m from dumpsite), ST2 = station 2 (200m away from dumpsite), ST3 = station 3 (300m away from dumpsite), ST4 = station 4 (400m away from dumpsite), ST5 = station 5 (500m away from dumpsite). Fig 6: Fungal Counts for Agbarho Air Samples
  • 5. Microbial Assessment of Air In The Vicinity Of Some Dump Sites in Delta State International organization of Scientific Research 11 | P a g e Key: DS = dumpsite, ST1 = station 1 (100m from dumpsite), ST2 = station 2 (200m away from dumpsite), ST3 = station 3 (300m away from dumpsite), ST4 = station 4 (400m away from dumpsite), ST5 = station 5 (500m away from dumpsite). Fig 7: Fungal Counts for Agbor Air Sample Key: DS = dumpsite, ST1 = station 1 (100m from dumpsite), ST2 = station 2 (200m away from dumpsite), ST3 = station 3 (300m away from dumpsite), ST4 = station 4 (400m away from dumpsite), ST5 = station 5 (500m away from dumpsite). 3.3 Bacterial Isolates from Air Samples The bacterial genera isolated from the air sample were Staphylococcus sp, Streptococcus sp, Micrococcus sp, Bacillus sp, Pseudomonas sp, Escherichia coli, and Klebsiella sp were isolated from the air samples (Table 1) Table 1:Identification and characterization of bacteria from air samples Isolate A B C D E F G Cultural characteristics Creamy converse colonies with smooth edges Cocci clusters Round clusters dented white flat & smooth colonies Dented and raised colonies Pink convex Smooth colonies Flat yellow swarming colonies Pigmentation Cream Cream Yellow Cream Green Pink cream Cellular morphology Cocci Cocci Cocci Rods Rods Rods Rods Gram Stain + + + + _ _ _ Aerobic growth + + + + + + + Anaerobic growth + + _ _ _ _ + Motility Test _ _ _ + + + + CatalaseTest + _ + + + + + OxidaseTest _ _ + _ + _ _ Urease Test + _ + _ + _ + Endospore Test _ _ _ + _ _ _ IndoleTest + _ _ _ _ + _
  • 6. Microbial Assessment of Air In The Vicinity Of Some Dump Sites in Delta State International organization of Scientific Research 12 | P a g e Key: + =positive, – =negative 3.4 Distribution of Bacterial Isolates in Air Samples The distribution of the bacterial isolates obtained from the air samples in wet and dry seasons are presented in Tables 2 and 3 respectively. It was observed that Streptococcus sp, Escherichia coli, Staphylococcus sp, Pseudomonas sp, Micrococcus sp, Klebsiella sp and Bacillus sp were more abundant in the air in the dumpsite and decreased with distanceaway from the dumpsites in the wet season while Micrococcus sp and Bacillus sp were abundant in both wet and dry seasons. Table 2:Distribution of air bacterial isolates in wet season Organism Dumpsite ST1 ST 2 ST3 ST 4 ST 5 Streptococcus sp +++ +++ ++ ++ ++ ++ Bacillus sp +++ +++ ++ ++ ++ ++ Escherichia coli +++ +++ ++ ++ ++ ++ Micrococcus sp +++ +++ ++ ++ ++ ++ Staphylococcus sp +++ +++ ++ ++ ++ ++ Pseudomonas sp. +++ +++ ++ ++ ++ ++ Klebsiella sp +++ +++ ++ ++ ++ ++ Key: – Absent, + Rare, ++ Intermediate, +++ large in number Table 3:Distribution of air bacterial isolates in dry season Key: – Absent, + Rare, ++Intermediate, +++large in number 3.5 Fungal Isolates from Air Samples The fungal isolates from the air, samples are presented in Table 4. The fungal genera isolated were Alternaria sp, Aspergillus sp, Cladosporium sp, Curvularia sp, Mucor sp, Penicillium sp and Rhizopus sp. Citrate Test + + + + + + _ Glucose Fermentation test + _ + + + + + Lactose Fermentation test _ _ _ _ _ + + H2S production test _ _ _ _ _ _ _ Organism identified Staphylococcus sp Streptococcus sp Micrococcus sp Bacillus sp Pseudomo nas sp E. coli. Klebsiella sp Organism Dumpsite ST1 ST 2 ST3 ST 4 ST 5 Streptococcus sp ++ ++ ++ ++ + + Bacillus sp. +++ +++ +++ +++ ++ ++ Escherichia coli ++ ++ ++ ++ + + Micrococcus sp +++ +++ +++ +++ ++ + Staphylococcus sp ++ ++ ++ ++ + + Pseudomonas sp. ++ ++ ++ + + + Klebsiella sp ++ ++ ++ + + -
  • 7. Microbial Assessment of Air In The Vicinity Of Some Dump Sites in Delta State International organization of Scientific Research 13 | P a g e 3.6 Distribution of Fungal Isolates in Air Samples The distribution of the fungal isolates obtained from the air samples in wet and dry seasons are presented in Tables 5 and 6 respectively. It was observed that all the fungal isolates were abundant in the dry season while Aspergillus sp and Mucor sp were abundant in both wet and dry seasons. Table 5:Distribution of air fungal isolates in wet season Organism Dumpsite ST1 ST2 ST3 ST4 ST 5 Aspergillus sp +++ ++ ++ ++ ++ ++ Cladosporium sp ++ + + + + + Curvularia sp ++ + + + + + Mucor sp +++ ++ ++ ++ ++ ++ Rhizopus sp ++ + + + + + Penicillium sp ++ + + + + _ Alternaria sp ++ + + + + + Key: – Absent, + Rare, ++Intermediate, +++large in number Table 6:Distribution of air fungal isolates in dry season Organism Dumpsite ST1 ST2 ST3 ST4 ST 5 Aspergillus sp +++ ++ ++ ++ ++ ++ Cladosporium sp +++ ++ ++ ++ ++ ++ Curvularia sp +++ ++ ++ ++ ++ ++
  • 8. Microbial Assessment of Air In The Vicinity Of Some Dump Sites in Delta State International organization of Scientific Research 14 | P a g e Mucor sp +++ ++ ++ ++ ++ ++ Rhizopus sp +++ ++ ++ ++ ++ ++ Penicillium sp +++ ++ ++ ++ ++ ++ Alternaria sp +++ ++ ++ ++ ++ ++ Key: – Absent, + Rare, ++Intermediate, +++large in number IV. DISCUSSION The microbial loads of the air samples taken from the dumpsites were higher than the normal atmospheric concentration of the microorganisms as the reported average level of the microbes in the ambient air is 3.0 log10cfu/ml [15]. This is an indication of the extent of microbial pollution of waste dump sites in the study sites The microorganisms isolated from the air samples taken from the various locations (Warri, Agbarho and Agbor), and distances sampled were similar in composition and distribution with those isolated from the air samples taken from the dumpsite. Generally the microbial load decreased with distances away from the dumpsite. The bacterial load for the air samples from Warri, Agbarho and Agbor presented in Figs 2- 4 showed that the bacteria counts decreased with distance from the dumpsite. It was equally observed that the bacterial counts were higher in the months of July August and September (wet season) and decreased slightly in the months of October, November and December (dry season). The decreasing bacterial counts with distance away from the dumpsite could be due to increased microbial activity in the dumpsite as a result of putrefaction and increased decomposition of organic matter in the vicinity of the dumpsite. It could also be as a result of household and industrial wastes deposited in the waste dumpsites as well as contaminants generated naturally that were propelled through the air, such as particles of dust and soil microbial spores in the air within the dumpsites. These results agree with the report of McCarthy [16], who listed these amongst others as possible sources of air contaminants. The observed increased trend in the bacterial counts in wet season could be as a result of increased rainfall in the months of July, August and September and decreased rainfall in the months of October, November and December leading to reduced water activity. These results agree with the reports of Obire et al. [17], who stated that seasonal variations favour physiological types. The fungal load for the air samples from Warri, Agbarho and Agbor as presented in Figs 5-7 showed that the fungal counts decreased with distance away from the dumpsites. It was equally observed that the fungal counts were lower in the months of July August and September (wet season) and increased slightly in the months of October, November and December (dry season). The decreasing fungal count with distance away from the dumpsite could be due to same reasons propounded for bacterial counts above. These results agreed with the report of McCarthy [16], who reported similar suggestions The observed trend in the fungal counts could be due to spore formation due to increased rainfall in the months of July, August and September and decreased rainfall in the months of October, November and December. These findings also agree with the reports of earlier researchers [17] who opined that seasonal variations favour physiological types. The bacterial and fungal genera encountered in this study had been reported by previous workers [18- 20], as the possible microbial isolates from the air. The isolated bacteria were more abundant in wet season than in the dry season. This could possibly be due to the atmospheric particles to which the microbes are attached being deposited by the process of rainfall. The increased water activity therein provides favourable conditions for bacteria to thrive and multiply. Bacillus sp and Micrococcus sp were more abundant in the air sample both in wet and dry seasons. The presence and prevalence of some of these species of bacteria in the dumpsite could be as a result of the presence of damp organic materials, materials impregnated with water, food and food products and spores of microorganisms propelled through the air. These results agree with the report of Osha [21] and Sola [22], who reported these as possible sources of air microflora. These bacteria, according to Douwes [23] can cause different forms of bacterial pneumonia, influenza and gastrointestinal diseases. The general trend of the isolated fungi showed that they were more abundant in dry season than in the wet season. This could be as a result of various climatic and local topographic factors including reduced rainfall leading to reduced water activity which favour germination of spores.It could also be as a result of the wind action and harsh weather conditions experienced during the dry season. Cladosporium sp, Penicillium sp, Mucor sp, Alternaria sp, Aspergillus sp, Rhizopus sp and Curvularia sp were observed to be predominant in the air samples during the dry season while Aspergillus sp and Mucor sp species were predominant in both the wet and
  • 9. Microbial Assessment of Air In The Vicinity Of Some Dump Sites in Delta State International organization of Scientific Research 15 | P a g e dry season. These results agree with the report of Prescott et al. [24], who listed these amongst the most common allergenic moulds associated with man and live stocks. Obire et al. [17] also identified Aspergillus sp as one of the most common fungi. The present study has revealed that the dumpsite had impact on both the microbial load and quality of such environment. Also, the microbial load of air in dumpsites decreases with increase in distance from the dump sites. Airborne bacteria at dumpsites are more prevalent in wet seasons than in the dry seasons, the reverse being the case for airborne fungi. It is therefore recommended that dumpsites should be located at about I km away from residential areas to avoid contamination of the air. Additionally, educational programmes should be organized for public awareness of impact of indiscriminate dumping of refuse. REFERENCES [1] F. C. Akharayi and F.O. Omoya. Microflora of Refuse Dumps and sites in five localities in Ondo state, Nigeria. Inter. J. Environ. 2(1-2), 2004, 27-32. [2] R. Madhukar and S. Srikantaswamy. Assessment of Air Quality in Bidadi Industrial Area, Ramanagaram District, Karnataka, India. J. Environ. Sci., Comp. Sci. Eng. Technol. 2(4), 2013, 1135-1140. [3] A. A. Jaffal, H. Nsanze, A. Benar, A. S. Ameen, I. M. Banat and A. A. EL-Moghett. Hospital airborne microbial pollution in a desert country. Environ. Inter., 1997, 23(2):67-172. [4] T. T. Adebolu and K. J. Vhriterhire. Survey of the microbial flora of the Ondo State Specialist Hospital Environment, Akure, Nigeria. Nig. J. Microbiol., 16(112), 2002, 91-94. [5] J. Dutkiewicz and M. Augustowska. Variability of airborne microflora in a hospital ward with a period of one year. Ann. Agric. Environ. Med., 13, 2006, 99-106. [6] D. H. Tambeker, P. B. Gulhane and D. D. Bhakare. Studies on environmental monitoring of microbial air flora in hospitals. J. Med. Sci. 7(1), 2007, 67-72. [7] F. O. Ekhaise, O. U. Ighosewe and O. D. Ajakpori. Hospital indoor airborne microflora in private and government owned hospitals in Benin City, Nigeria. World. J. Med. Sci., 3(1), 2008, 34-38. [8] S. Sudharsan, P. Srikath, M. Sheela and R. Stienberg. Study of indoor air quality in hospitals in South Chennai, India. Ind. Blt. Environ. 17, 2008, 435-441 [9] F. O. Ekhaise, E. E. Isitor, O. Idehen, and A. O. Emoghene. Airborne Microflora in the Atmosphere of an Hospital Environment of University of Benin Teaching Hospital (UBTH), Benin City, Nigeria. World J. Agri. Sci., 6(2), 2010, 166-170 [10] S. A. Awosika, F. A. Olajunbu, and N. A. Amusa. Microbiological assessment of indoor air of a teachinghospital in Nigeria. Asian Pac. J. Trop. Bio. 16, 2012, 465-468. [11] M. N. O. Omoigberale, O. O. Amengialue and M. I. Iyamu. Microbiological assessment of Hospital Indoor Air Quality in Ekpoma, Edo State, Nigeria. Global Res. J. Microbiol., 4(1), 2014, 1-5. [12] M. Cheesebrough. District Laboratory Practice in Tropical Countries. Part 2, 2000, 32-33. [13] M. Cheesbrough. District Laboratory Practice In Tropical Countries, part 2, second edition Cambridge University press, The Edinburgh Building, Cambridge, United Kingdom, 2006, 38, 62-69. [14] W. F. Harrigan, and M. E. McCance. Laboratory Methods in Food and Dairy Microbiology. 8th Edition Academic press. London.1990, 210 [15] Y. Panthi, and U. Shrestha. Microbial Air Pollution A case study of sodisol sanitary landfill site. Asian J. Appl. Sci. 1(1), 2008, 94-97. [16] J. McCarthy. Indoor Air Quality Handbook. Mc-Graw Hill London. 2001, 290. [17] O. Obire. A. O. Nwaubeta and S. B. N. Adue. Microbial Community of a Waste Dumpsite. J. Appl. Sci. Environ. Manag. 2, 2002, 73-82. [18] D. K. G. Ayanru. Global Impact Air Spore. Appl. Microbiol. 6, 1981, 107-117. [19] K. J. Ryan, C. G. Ray and S. Sherris. Medical Microbiology. 4th edition: McGraw Hill, New York. 2004, 623-625. [20] P. M. Kirk, P. F. Cannon, D. W. Minter and J. A. Stalpers. Dictionary of the Fungi .10th ed. Wallingford: 2008, 222. [21] S. Osha. Microbial contamination of the Environment. J. Curr. Sci. 89, 1999, 101-109 [22] G. Sola. Indoor Air Quality. J. Appl. Environ. Microbiol. 65, 2000, 423-432. [23] J. Douwes. Bioaerosol health effects and exposure assessment: Progress and prospects. Ann. Occup. Hyg., 47(3), 2003, 187-200. [24] L. M. Prescott, J. P. Harley, and A. D. Klein. Microbiology. 6th Edition. McGraw-Hill, New York. 2005, 962 - 965