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Drinking Water Distribution Systems of Dams in Ondo State, Nigeria as Reservoir of Multi-Drug Resistant Bacteria

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The occurrence of multidrug resistant (MDR) bacteria across food, water and environment is a major global public health challenge. Drinking water distribution systems of selected dams in Ondo State Nigeria was investigated for their physicochemical properties, microbial quality and presence of MDR bacteria. It was observed that the pH of the water ranged between 5.20 to 7.40 while DO ranged between 1.45 to 6.82mg/l and the residual chlorine of the water was between 0.00 to 0.09mg/l. Coliform count of the water samples range from 0 to 9.33 x10 CFU/ml. Gram negative bacteria recovered from the water include Proteus, klebsiella, Alcaligenes, 2 Pseudomonas etc. and they were observed to show multiple resistant to various antibiotics which include tetracycline, streptomycin, ampicillin, sulfamethoxazole. This study revealed that MDR bacteria of public health significance are present in the drinking water distribution system of Ondo State, Nigeria.
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World Applied Sciences Journal 32 (3): 403-414, 2014
ISSN 1818-4952
© IDOSI Publications, 2014
DOI: 10.5829/idosi.wasj.2014.32.03.83237
Corresponding Author: A.T. Adesoji, Department of Biological Sciences, Federal University Dutsin-Ma, Katsina State, Nigeria.
403
Drinking Water Distribution Systems of Dams in Ondo State,
Nigeria as Reservoir of Multi-Drug Resistant Bacteria
A.T. Adesoji, A.A. Ogunjobi and I.O. Olatoye
1,2 2 3,4
Department of Biological Sciences, Federal University Dutsin-Ma, Katsina State, Nigeria
1
Department of Microbiology, University of Ibadan, Nigeria,
2
Department of Veterinary, Public Health and Preventive Medicine, University of Ibadan, Nigeria
3
Paul G. Allen School for Global Animal Health, Washington State University, USA
4
Abstract: The occurrence of multidrug resistant (MDR) bacteria across food, water and environment is a major
global public health challenge. Drinking water distribution systems of selected dams in Ondo State Nigeria was
investigated for their physicochemical properties, microbial quality and presence of MDR bacteria. It was
observed that the pH of the water ranged between 5.20 to 7.40 while DO ranged between 1.45 to 6.82mg/l and
the residual chlorine of the water was between 0.00 to 0.09mg/l. Coliform count of the water samples range from
0 to 9.33 x10 CFU/ml. Gram negative bacteria recovered from the water include Proteus,klebsiella,Alcaligenes,
2
Pseudomonas etc. and they were observed to show multiple resistant to various antibiotics which include
tetracycline, streptomycin, ampicillin, sulfamethoxazole. This study revealed that MDR bacteria of public health
significance are present in the drinking water distribution system of Ondo State, Nigeria.
Key words: Multidrug resistant bacteria Antibiotics Dams Water distribution systems
INTRODUCTION (which involves direct cell-to-cell contact and transfer of
Water is an essential natural resource for the uptake of free DNA that results from bacterial lysis
sustainability of life on earth [1]. One of the targets of the [10, 11].
millennium development goals (MDG) in terms of healthy Water distribution system can therefore, be a source
living for the masses can be achieved through the supply of transfer of antibiotic resistance genes between bacteria
of safe and convenient water [2]. The presence of isolates which eventually can result into public health
pathogenic (disease-causing) organisms (bacteria) is a threat which can result into high cost and long period of
concern when considering the safety of drinking water. treatment. However, to the best of our knowledge no
[3]. However, many authors have reported the isolation of study has been carried out on the occurrence of antibiotic
bacteria like Aeromonas,Methlobacterium,Bacillus and resistant bacteria in water distribution systems of dams in
Pseudomonas from diverse aquatic environments like well Ondo State, Nigeria. This study therefore, aims at
water and heavily polluted waters [4-6]. determining if drinking water distribution systems of
However, studies have shown increased level of selected dams located in Ondo State, Nigeria would be a
bacteria resistance to various antibiotics among these source of multidrug resistance bacteria.
water bodies bacteria due to misuse of antibiotics in
human health, veterinary, aquaculture, agriculture and MATERIALS AND METHODS
household products [7,8]. Whenever antimicrobials are
used, bacteria inevitably develop resistance mechanisms Study Areas: Two water distribution systems which
either through spontaneous mutations or by acquiring include Owena-Ondo and Owena-Ijesha were selected in
genes from other bacteria. The later may occur by Ondo State, Nigeria for this study. Both dams were
transduction (mediated by bacteriophages); conjugation constructed and owed by Ondo State government.
plasmids or transposons); or transformation, involving
mg of Solids in the beaker x 1000
Total solid, TS (mg/l) = Volume of sample
World Appl. Sci. J., 32 (3): 403-414, 2014
404
Owena-Ijesha dam was commissioned in 1965 with an picked and streaked out on Nutrient Agar plate for
installed capacity of 19600 m /day and to supply water to purification. Cultures were then stored at 4°C on Nutrient
3
a population of 787867 people. Owena-Ondo dam was Agar (NA) slant [13].
commissions in 1971 and installed with a capacity of
5450 m /day and designed for an estimated populace of Physicochemical: The pH of the water samples was
3
192340. The dam is supplied with water from the Owena determined by the use of pH meter while Biological
River and it covers an appropriate surface area of 7.8km . Oxygen demand (BOD), Chemical Oxygen demand (COD),
2
Sample Collection and Analysis: Water samples for was determined by the method of Skoog and West [14]
microbiological and physicochemical analysis were and Radojeric and Baskin [15]. DO was determined by
obtained two times each between December, 2010 and measuring 200cm of water sample into a beaker with
January, 2011 to represent onset of dry season and June, measuring cylinder. A probe of the DO was then inserted.
2011 and July, 2011 to represent the start of raining The DO meter was switched on and the DO value (mg/l)
season from raw, treated and two municipal taps of the was recorded after 2 minutes of automated value
water distribution systems. Raw untreated water and adjustment. Conductivity of the water was determined by
treated water of each of the dams were only analyzed for inserting a conductivity electrode in water samples while
physicochemical analysis. The following steps were residual chlorine of water samples were determined by the
followed when sampling water for microbial analysis as use of a chlorine comparator with N,N-
previously described by Rice [12]. Plastic sample bottles diethylparaphenylenediamine (DPD)
(50ml) were used for the sampling. Samples for the treated Total solid (TS) was determined by keeping the water
water i.e. dams final output and municipal taps was taken samples at 103°C in a clear dry glass beaker of 150ml
by opening of the taps and allowing the water to run for capacity in an oven for 1 hr. The capacity and appropriate
3-4min before collection. Collected samples were then kept identification mark was then place on it. After, 100ml of
at 4°C in the cooler box packed with ice and transported the thoroughly mixed sample was measured into the
to the laboratory for analysis within six hours. Samples beaker. The beaker was then placed in an oven maintained
from the raw water were taken from the dam output before at 103°C for 24 hrs. After 24 hours, the beaker was then
getting to the treatment plants. Samples for chemical cooled and weigh. The weight of the solid in the beaker
analysis were taken with 500ml sample bottles from the was determined by subtracting the weight of the clean
raw water and treated water of the dams and then beaker from the weight determined after addition and
transported into the laboratory for chemical analysis. drying of the sample in the beaker. Total solid (TS) was
Microbial: Serial dilution of the water samples was carried
out aseptically up to 10 in order to obtain countable
4
bacteria colonies on the agar plate. The samples were then
mixed by shaking before plating on appropriate media.
Total plate counts was determined by plating out with a Total dissolved solid (TDS) was determined as TDS
sterile pipette 1ml of the diluted samples from 10 and (mg/l) = mg of solid in the beaker x 1000 (volume of
2
10 into sterile petri dish and Nutrient agar that has been sample) while
4
sterilized and kept at 45°C in a water bath was aseptically Total suspended solid (TSS) was determined as TSS
poured into the petri dish and allowed to solidify on the (mg/l) = TS (mg/l) TDS (mg/l)
flat surface. The plates were then incubated in an inverted
position in an incubator set at 37°C overnight. Colonies Molecular Characterization of Bacteria Using16s rDNA
developed on the agar plates were then counted with a Sequencing: Total genomic DNA was extracted from
colony counter. For all treated water samples i.e. the final isolates after streaking stock culture on Luria Betani (LB)
dam water samplings and the municipal samplings, agar overnight followed by dispensing 200 µl of 5% chelex
undiluted samples of the water and samples diluted to in a tube and then taking a loopful of a colony of
10 was plated out on the Nutrient agar (NA) plates. bacterium into the chelex solution. Mixture of the bacteria
1
Similar steps were also carried out for coliform on Eosin and chelex solution was then boiled at 100°C for 10 min
methylene blue (EMB) agar and deoxcholate agar which and centrifuged at 13k x g for 1min. Extracted DNA
was used as selective agar for isolation of E. coli and supernatant (5µl) was used as template with 2mM MgCl ,
other coliform. Colonies with different morphologies were 0.8 mM dNTPs, 0.2 µM of each primer 1 and primer 2 and
Dissolve Oxygen (DO) and Total Organic carbon (TOC)
3
then determined as follows:
2
World Appl. Sci. J., 32 (3): 403-414, 2014
405
Table 1: Antibiotic concentration used for breakpoints
Antibiotics for gram negatives with concentration (ug/ml) Antibiotics for gram positives with concentration (ug/ml)
FF Florfenicol (16) SU Sulfamethoxazole (512)
T Tetracycline (16) AM Ampicillin (0.5)
S Streptomycin (16) T Tetracycline (16)
G Gentamycin (16) SXT Sulfamethoxazole/Trimethoprim (76/4)
K Kanamycin (64) G Gentamycin (16)
C Chloramphenicol (32) E Erythromycin (8)
N Nalidixic Acid (30) RIF Rifampin (4)
AMC Amoxillin/Clavulanic Acid (32/16) LIN Lincomycin (4)
CEF Ceftiofur (12) CIP Ciprofloxacin (4)
SU Sulfamethoxazole (512)
SXT Sulfamethoxazole/Trimethoprime (76/4)
1X PCR buffer. Reaction condition included 1min solid (TDS) of both the raw and untreated water was
denaturation (95°C) followed by 30 cycles of 96°C for 30s, observed not to be above the 500 mg/l limit recommended
60°C for 30s and 72°C for 30s and a final extension of 72°C by WHO. [20]. Dissolved oxygen (DO) of both the raw
for 10min. PCR products were then separated and and untreated water ranged between 1.45 to 3.11 mg/l and
visualized on 1% agarose gel electrophoresis to confirm 4.33 to 6.82 mg/l respectively. These values exceed the
amplification. The 16s rDNA sequence was amplified range of 2.5 to 3.8 mg/l observed by Fatombi et al. [21] in
using 16s-8F (AGAGTTTGATCMTGGCTCAG) and their study with surface water in Lagbe town. But these
16s-517R (ATTACCGCGGCTGCTGG) primers [16, 17]. values obtained in this study were below WHO
PCR products were sequenced (Eurofins MWG, USA) and recommendation. As suggested by WHO [22] the
manual base calls were sequence trimming was completed conductivity of portable water should not exceed 300
by sequencher (5.0). BLASTn was used to identify closes µs/cm. In this study, we obtained the conductivity range
sequence matches (www.ncbi.nlm.nib.gov/BLAST/blast) of 32.50 to 42.00 µs/cm and 5.6 to 59.50 µs/cm in raw and
Antibiotic Breakpoint Susceptibility Test: The antibiotic Also in this study, no residual chlorine was detected in
resistance profile of the bacteria was determined using the raw water samples, but the treated water has values of
breakpoint assays on LB agar plates. The Agar medium 0.03 mg/l and 0.09 mg/l at Owena-Ondo dam during dry
was autoclaved, cooled to 45°C and then antibiotics were and wet season sampling periods respectively. These
added to specific concentration (Table 1) before pouring values were below 0.5 mg/l maximum limit recommended
the medium into petri dishes (150 x 15mm). Overnight by WHO. In this study, we observed that the Total solid
cultures were then ‘stabbed’ from the 96-well plate onto (TS) and Total suspended solid of the raw water sampling
agar plates using 96-well pin replicator and incubated ranged between 190 to 430 mg/l and 120 to 370 mg/l
overnight at 37°C. Isolates were scored as ‘1’for growth respectively. These values were observed to be higher
of ‘0’ for no growth on each antibiotic plate. than values obtained for the treated water which ranges
RESULTS AND DISCUSSION respectively. This showed that the treatment process was
Physicochemical Properties of Water: The results of the passed through the treatment plant. Also, we observed
physicochemical properties of the raw and treated water that the Total dissolve solid (TDS) of the raw water was
are shown on Table 2 and 3 respectively. The results between 50 mg/l to 66 mg/l compared to higher range
obtained showed that the pH of the raw water during the observed for the treated water samples (83.00 to 100 mg/l).
two seasons of sampling ranged between 6.50 and 7.50 in The values of TDS and TSS recorded in this study are
raw water and 5.20 and 7.70 in treated water. The pH of the below WHO recommendation.
raw water was similar to the result of Pranavam et al. [18].
Ravisankar and Poongothai. [19] described such water pH Microbial Quality of the Water Samples: The results of
as either sharply acidic or alkaline in nature. However, the the coliform and total plate counts of the water samples
minimum range of the pH obtained for the treated water from the water distribution system of these dams are
during the wet and dry seasons sampling in both shown on Table 4. We observed that the coliform and
locations- (Owena-Ondo and Owena-Ijesha) was total bacteria count was higher in raw water compared to
observed to be slightly acidic in nature. Total dissolved the treated water in some of the water distribution system
treated water respectively during both sampling periods.
between 93.00 mg/l to 320 mg/l and 9.20 to 210 mg/l
effective in reduction in the solid material in the water as
World Appl. Sci. J., 32 (3): 403-414, 2014
406
Table 2: Physicochemical properties of raw water sampling of Owena-Ondo and Owena-Ijesha dams in Ondo State, Nigeria during dry and raining season of December, 2010 and June, 2011
pH BOD (mg/l) COD (mg/l) DO (mg/l) TOC (mg/l) TDS (mg/l) TS (mg/l) TSS (mg/l) Conductivity (µs/cm) Residual Chlorine(mg/l)
Owena-Ondo June/July 6.50 4.83 53.60 3.11 6.22 66.00 420.00 3 50.00 42.00 0.00
December/January 7.40 6.83 49.80 1.45 1.34 62.00 190.00 120.00 34.00 0.00
Owena-Ijesha June/July 7.10 6.62 64.60 3.21 6.43 50.00 430.00 370.00 32.50 0.00
December/January 7.30 5.72 56.10 2.34 1.43 58.00 360.00 287.00 34.80 0.00
WHO limit 6.5-8.5 6-9 - - - 500 - 500 300 0.5
Table 3: Physicochemical properties of treated water sampling of selected dams in Ondo State, Nigeria during dry and raining season of December, 2010 and June, 2011
pH BOD (mg/l) COD (mg/l) DO (mg/l) TOC (mg/l) TDS (mg/l) TS (mg/l) TSS (mg/l) Conductivity (µs/cm) Residual Chlorine(mg/l)
Owena-Ondo June/July 5.40 1.32 47.40 5.21 1.65 80.00 220.00 1 30.00 40.00 0.09
December/January 7.70 2.11 81.30 4.83 1.90 96.00 130.00 32.80 59.50 0.03
Owena-Ijesha June/July 7.40 3.11 34.80 4.33 2.45 100.00 320.0 210.00 52.00 0.04
December/January 5.20 1.87 78.00 6.82 1.75 83.00 93.00 9.20 5.60 0.05
WHO limit 6.5-8.5 6-9 - - - 500 - 500 300 0.5
Table 4: Coliform and Total plate counts of Owena-Ondo and Owena-Ijesha water distribution systems during raining and dry seasons sampling
Coliform count (CFU/ml) Total bacteria count (CFU/ml)
---------------------------------------------------------------------------------------------- ---- ------ ------ ------- ------ ------- ------ -----------------------------------------------------
June/July December/January June/July December/January
---------------------------------------------- ---------------------------------------------- ------------------------- --------------------- -----------------------------------------------
Owena-Ondo Owena-Ijesha Owena-Ondo Owena-Ijesha Owena-Ondo Owena-Ijesha Owena-Ondo Owena-Ijesha
Raw water 5.6 x 10 3.02 x 10 3.2 x 10 4.0 x 10 3.2 x 10 3.3 x 10 3.2 x 10 6.0 x 10
2 5 53 3 55
Treated water 3.0 x 10 2.0 x 10 4.8 x 10 3.0 6.2 x 10 6.4 x 10 6.8 x 10 0
22
Municipal Tap 1 0 8.91 9.33 x 10 0.0 0 4.2 x 10 2.0 x 10 0
25
Municipal Tap 2 0 8.91 2.13 x 10 4.0 x 10 0 1.4 x 10 4.0 x 10 2.0 x 10
22 2 2
sampled (e.g. Owena-Ijesha). However, the count in some of Owena-Ondo and Owena-Ijesha respectively (Table 5).
of the treated water exceeded what was suggested by It was observed that Bacillus was the highest occurred
WHO [22] For drinking water of no coliform per 100ml. bacteria in the entire sample points of Owena-Ondo dam.
For example, treated water and the water from municipal Bacillus species are Gram positive aerobic spore-forming
taps of Owena-Ijesha during June/July sampling has and most members of the genus are saprophytic prevalent
coliform count ranging between 8.91 to 2 x 10 CFU/ml. in the soil, water and air and on vegetation [25]. However,
2
During dry season, samples obtained at the same dam high occurrence of this Bacillus particularly in the treated
showed values ranging from 3.0 to 4.0 x 10 CFU/ml while municipal water could be as a result of spores formation
2
no coliform count was obtained during wet season that makes them of very high resistance to chlorine
sampling of both municipal taps of Owena-Ondo dam. disinfectant used in the water treatment plant. At Owena-
During December/ January sampling at Owena-Ondo Ijesha Proteus,Pseudomonas and Kebsiella showed the
coliform bacteria count range between 4.8 x 10 CFU/ml at highest percentage of bacteria from the raw water and
their treated water to 9.33 x 10 at the first municipal tap each constitute 5% of total bacteria while Bacillus was
2
sampled while its total plate count ranges between 6.8 x 10 the highest at the treated water and municipal 2 tap
CFU/ml at the treated water to 2.2 x 10 CFU/ml at the raw constituting 60% and 41.67% of the total bacteria. Other
5
water sample. However, EPA [23] reported that isolation bacteria isolated in these treated and municipal water
of coliform from water is of great importance in different include Ralstonia, Proteus, Alcaligenes, Acinetobacter,
fields of microbiology as is used as a continuous standard Citrobacter, Enterobacter, klebsiella, Pseudomonas and
when water is under study. They are described as Enterococcus which were similar to bacteria reported by
important indicator of water quality and along with other Suthar et al. [26]. These authors isolated a wide range of
organism; they make up an important part of water pathogenic bacteria from potable water samples from
standard. In this study, the range of coliform count some rural habitation of Northern Rajasthan, India.
obtained in some of the water samples correlate with the The presence of some of these coliforms is an indication
coliform with the coliform count range of up to 4.7 x 10 to of fecal pollution and a consequent hazard of contracting
8.0 x10 CFU/ml reported by Oluyege et al. [24], in treated water-borne disease from the use of such water [27].
and untreated water samples from Ero dam in Ekiti State Such polluted water could be as a result of leakage of
Nigeria. pipes conveying this water to the final consumers and
Bacteria Identified, There Susceptibility to Antibiotics Some of these bacteria like Proteus,Klebsiella and
and MDR Bacteria: In this study, a total of 72 and 61 Pseudomonas etc. have been described as opportunistic
bacterial isolate were recovered from the sample locations pathogens [28]. This occurrence of these bacteria in some
ineffectiveness of water treatment plant processing.
World Appl. Sci. J., 32 (3): 403-414, 2014
407
Table 5: Bacteria isolated from Owena-Ondo (Dam 5) and Owena-Ijesha (Dam 6) water samples as Identified by 16S rDNA sequencing
Sampled Dam Location Bacteria No of Isolates Percentage total bacteria from location (%)
Owena Ondo Raw water Bacillus sp 7 53.85
Escherichia sp 1 7.69
Klebsiella sp 3 23.08
Leucobacter sp 1 7.69
Uncultured bacteria clone 1 7.69
Total bacteria (Raw water) 13
Treated water Alcaligenes sp 2 18.18
Aquitalea sp 1 9.09
Bacillus sp 3 27.27
Klebsiella sp 1 9.09
Morganella sp 1 9.09
Pseudomonas sp 1 9.09
Proteus sp 1 9.09
Staphylococcus sp 1 9.09
Total (Treated water) 11
Municipal Tap 1 Alcaligenes sp 2 9.09
Bacillus sp 10 45.45
Escherichia sp 2 9.09
Lysinibacillus sp 2 9.09
Proteus sp 2 9.09
Pseudomonas sp 2 9.09
Providencia sp 2 9.09
Total (Municipal Tap 2) 22
Municipal Tap 2 Acinetobacter sp 1 3.85
Bacillus sp 13 50
Lysinibacillus sp 1 3.85
Morganella sp 4 15.38
Myroides sp 1 3.85
Proteus sp 5 19.23
Serratia sp 1 3.85
Total 26
Total bacteria from Owena-Ondo (72)
Table 5 (Cont’d): Bacteria from Owena-Ondo (Dam 5) and Owena-Ijesha (Dam 6) water samples as Identified by 16S rDNA sequencing
Owena-Ijesha Raw water Alcaligenes sp 4 12.50
Acinetobacter sp 2 6.25
Aeromonas sp 1 3.12
Morganella sp 1 3.12
Proteus sp 5 15.63
Providencia sp 2 6.25
Pseudomonas sp 5 15.63
Klebsiella sp 5 15.63
Serratia sp 1 3.12
Myroides sp 1 3.12
Bacillus sp 3 9.38
Lysinibacillus sp 1 3.12
Uncultured bacterium clone 1 3.12
Total (Raw water) 32
Treated water Ralstonia sp 1 20.00
Proteus sp 1 20.00
Bacillus sp 3 60.00
Total (Treated water) 5
Municipal Tap 1 Alcaligenes sp 2 16.67
Acinetobacter sp 1 8.33
Bacillus sp 2 16.67
Citrobacter sp 2 16.67
Enterobacter sp 1 8.33
Klebsiella sp 1 8.33
Pseudomonas sp 1 8.33
Enterococcus sp 2 16.67
Total 12
Municipal Tap 2 Alcaligenes sp 2 16.67
Bacillus sp 5 41.67
Brevundimonas sp 1 8.33
Chromobacterium sp 1 8.33
Proteus sp 1 8.33
Pseudomonas sp 1 8.33
Lysinibacillus sp 1 8.33
Total 12
Total bacteria from Owena-Ijesha (61)
World Appl. Sci. J., 32 (3): 403-414, 2014
408
Table 6: Multidrug Resistant bacteria from Owena-ondo water samples and their resistance phenotypes (Dam 5)
Sources Bacteria/Strain ID Resistant Phenotypes
Raw water (Gram negatives)
OWODRW Uncultured bacterium clone (230A) AM, SU, GEN
OWODRW Klebsiella pneumoniae (335) AM, SU, SXT, AMC, C, CEF, FF
Raw water (Gram positives)
OWODRW Bacillus cereus (338) SU, E, RIF, LIN
OWODRW Bacillus sp. (K) AM, SU, SXT, LIN
OWODRW Bacillus cereus (232) AM, SU, SXT, LIN
OWODRW Leucobacter komagatae (230B) AM, SU, SXT, GEN
Treated water (Gram negatives)
OWODFW Pseudomonas otitidis (350) AM, SU, SXT
OWODFW Proteus mirabilis (201) AM, SU, SXT, S, CEF
OWODFW Alcaligenes sp (198) T, AM, SU, SXT, S, K
OWODFW Morganella morganii (199) AM, SU, AMC, S, CEF
OWODFW Alcaligenes faecalis (197) T, AM, SU, SXT, S, K, CEF
OWODFW Klebsiella sp (386B) T, AM, SU, SXT, AMC, S, C, FF
Treated water (Gram positives)
OWODFW Bacillus sp. (200A) AM, SU, SXT, LIN
OWODFW Bacillus sp. (202B) AM, SU, SXT, LIN
OWODFW Staphylococcus sp (202) AM, SU, SXT, T, GEN
Municipal Tap 1 (Gram negatives)
OWODM1 Proteus mirabilis (273) T, SU, SXT, S
OWODM1 Pantoea agglomerans (214) T, AM, SU, AMC, S
OWODM1 Alcaligenes sp. (272B) T, AM, SU, SXT, S, GEN, K
OWODM1 Morganella morganii (215A) T, AM, SU, SXT, AMC, S, CEF
OWODM3 Proteus vulgaris (372) T, AM, SU, SXT, S, C, N, CEF, FF
OWODM1 Pseudomonas sp (260) T, AM, SU, SXT, AMC, C, N, CEF, FF
Gram negative Antibiotics: Ampicillin (AM); Ceftiofur (CEF); Chloramphenicol and Florfenicol (FF); Kanamycin (K), Streptomycin (S) and Gentamycin
(GEN); Tetracycline (T); Nalidixic Acid (N); Sulfamethoxazole (SU); Sufamethoxazole/ Trimethoprim (SXT); Amoxicillin/Clavulanic Acid (AMC)Gram
positive Antibiotics: Sufamethoxazole (SU); Ampicillin (AM); Tetracycline (T); Gentamycin (GEN); Erythromycin; Riframprim (RIF); Lincomycin (LIN);
Ciprofloxacin (CIP), Sulfamethoxazole/Trimethoprin (SXT)OWODFW: Owena-ondo treated water, OWODM1: Owena-ondo municipal 1
Table 6 (Cont'd): Multidrug Resistant bacteria from Owena-ondo water samples and their resistance phenotypes (Dam 5)
Sources Bacteria/Strain ID Resistant Phenotypes
Municipal Tap 1 (Gram positives)
OWODM1 Bacillus altitudinis (271A) AM, T, RIF, LIN
OWODM3 Bacillus sp. (269A) AM, SU, SXT, LIN
OWODM3 Bacillus sp. (188) AM, SU, SXT, LIN
OWODM3 Bacillus thuringiensis (270) AM, SU, SXT, LIN
OWODM1 Bacillus cereus (215B) AM, SU, SXT, E, LIN
OWODM1 Bacillus sp. (245A1) AM, SU, T, RIF, LIN, CIP, GEN
Municipal Tap 2 (Gram negatives)
OWODM2 Escherichia coli (210A) T, AM, SU, AMC
OWODM2 Providencia rettgeri (253B1) S, SU, SXT, AMC
OWODM2 Pseudomonas putida (251B) T, AM, SU, SXT, S, C,
OWODM3 Serratia marcescens (218A) T, AM, SU, AMC, CEF
OWODM3 Morganella morganii (218B) T, AM, SU, AMC, CEF
OWODM2 Escherichia coli (210B) T, AM, SU, SXT, AMC
OWODM3 Morganella morganii (169) T, SU, SXT, AMC, S, K, CEF
OWODM2 Alcaligenes faecalis (253A) T, AM, SU, SXT, S, GEN, K, C
OWODM3 Proteus vulgaris (190) T, AM, SXT, S, K, C, N, CEF, FF
OWODM2 Providencia rettgeri (209) T, AM, SU, SXT, S, C, N, CEF, AMC
OWODM2 Proteus vulgaris (184B) T, A M, SU, SXT, AMC, S, C, N, CEF, FF
OWODM3 Proteus vulgaris (171B) T, A M, SU, SXT, AMC, S, C, N, CEF, FF
Gram negative Antibiotics: Ampicillin (AM); Ceftiofur (CEF); Chloramphenicol and Florfenicol (FF); Kanamycin (K), Streptomycin (S) and Gentamycin
(GEN); Tetracycline (T); Nalidixic Acid (N); Sulfamethoxazole (SU); Sufamethoxazole/ Trimethoprim (SXT); Amoxicillin/Clavulanic Acid (AMC)
Gram positive Antibiotics: Sufamethoxazole (SU); Ampicillin (AM); Tetracycline (T); Gentamycin (GEN); Erythromycin; Riframprim (RIF); Lincomycin
(LIN); Ciprofloxacin (CIP), Sulfamethoxazole/Trimethoprin (SXT) OWODM2: Owena-ondo municipal 2 OWODM3: Owena-ondo municipal 3
World Appl. Sci. J., 32 (3): 403-414, 2014
409
Table 6 (Cont'd): Multidrug Resistant bacteria from Owena-ondo water samples and their resistance phenotypes (Dam 5)
Sources Bacteria/Strain ID Resistant Phenotypes
Municipal Tap 2 (Gram positives)
OWODM2 Bacillus sp (371) AM, SU, SXT, LIN
OWODM2 Bacillus sp (187) SU, SXT, AM, LIN
OWODM2 Bacillus sp. (245A2) AM, SU, SXT, LIN
OWODM2 Bacillus sp. (252A) AM, SU, SXT, LIN
OWODM2 Bacillus cereus (245B) AM, SU, SXT, LIN
OWODM2 Lysinibacillus sp (219*B) AM, SU, T, LIN, GEN
OWODM2 Bacillus pumilus (184A) SU, SXT, T, E, RIF, LIN
OWODM2 Bacillus cereus (186) AM, SU, SXT, T, E, RIF, LIN
OWODM2 Myroides odoratus (269B) T, AM, SU, SXT, AMC, S, GEN, K, CEF
Gram negative Antibiotics: Ampicillin (AM); Ceftiofur (CEF); Chloramphenicol and Florfenicol (FF); Kanamycin (K), Streptomycin (S) and Gentamycin
(GEN); Tetracycline (T); Nalidixic Acid (N); Sulfamethoxazole (SU); Sufamethoxazole/ Trimethoprim (SXT); Amoxicillin/Clavulanic Acid (AMC)Gram
positive Antibiotics: Sufamethoxazole (SU); Ampicillin (AM); Tetracycline (T); Gentamycin (GEN); Erythromycin; Riframprim (RIF); Lincomycin (LIN);
Ciprofloxacin (CIP), Sulfamethoxazole/Trimethoprin (SXT)OWODM2: Owena-ondo municipal 2OWODM3: Owena-ondo municipal 3
Table 7: Multidrug Resistant bacteria from Owena-Ijesha water samples and their resistance phenotypes (Dam 6)
Sources Bacteria/Strain ID Resistant Phenotypes
Raw water (Gram negatives)
OWIRW Klebsiella pneumoniae (346) T, AM, SXT, SU
OWIRW Klebsiella oxytoca (175A) AM, SU, SXT, K
OWIRW Proteus mirabilis (173A) T, SU, SXT, GEN
OWIRW Klebsiella sp. (347) T, AM, SU, SXT, S
OWIRW Providencia vermicola (227B) T, SU, S, GEN, CEF
OWIRW Morganella morganii (206) T, AM, SU, AMC, S
OWIRW Uncultured bacterium (205A) AM, SU, S, GEN, K
OWIRW Klebsiella sp (389B2) T, AM, SU, AMC, N
OWIRW Pseudomonas sp. (342A) AM, SU, AMC, CEF, FF
OWIRW Alcaligenes faecalis (173B) T, SU, SXT, AMC, S, CEF
OWIRW Serratia marcescens (348) T, AM, SU, AMC S, C, CEF
OWIRW Klebsiella pneumoniae (345) T, AM, SU, SXT, AMC, S, C
OWIRW Pseudomonas sp (223) SU, SXT, AMC, S, C, CEF, FF
OWIRW Acinetobacter baumannii (222) T, AM, SU, SXT, AMC, S, C, FF
OWIRW Pseudomonas sp (343B) T, AM, SU, AMC, S, N, CEF, FF
OWIRW Proteus vulgaris (179) T, SU, SXT, AMC, S, C, N, CEF, FF
OWIRW Pseudomonas sp (175B) T, SU, SXT, AMC, S, GEN, K, CEF
OWIRW Acinetobacter junii (205B) T, AM, SU, SXT, AMC, S, GEN, CEF
OWIRW Alcaligenes faecalis (207) T, AM, SU, SXT, AMC, S, K, N, CEF
OWIRW Myroides odoratus (174B) AM, SU, SXT, AMC, S, GEN, K, CEF
OWIRW Pseudomonas sp (343A) AM, SU, SXT, AMC, K, C, N, CEF, FF
OWIRW Proteus vulgaris (178B) T, AM, SU, SXT, AMC, S, C, N, CEF, FF
OWIRW Alcaligenes sp (174A) T, AM, SU, SXT, AMC, S, GEN, K, C, N, CEF
Gram negative Antibiotics: Ampicillin (AM); Ceftiofur (CEF); Chloramphenicol and Florfenicol (FF); Kanamycin (K), Streptomycin (S) and Gentamycin
(GEN); Tetracycline (T); Nalidixic Acid (N); Sulfamethoxazole (SU); Sufamethoxazole/ Trimethoprim (SXT); Amoxicillin/Clavulanic Acid (AMC)Gram
positive Antibiotics: Sufamethoxazole (SU); Ampicillin (AM); Tetracycline (T); Gentamycin (GEN); Erythromycin; Riframprim (RIF); Lincomycin (LIN);
Ciprofloxacin (CIP), Sulfamethoxazole/Trimethoprin (SXT)OWODM3: Owena-ondo municipal 3
of this drinking water rendered it unsafe for human acid and gentamycin. We observed among gram negative
consumption and can cause infections and diseases in bacteria from Owena-Ondo dam (Fig. 1) that out of the
immune-compromised individuals such as the old and total bacteria from each of the sample location between
infant which immune systems is still developing [29]. 0-100%, 0-78%,20-78%, 29-89%, 86-100%, 20-86% showed
The antibiotic resistance profile of these bacteria resistance to tetracycline, streptomycin, combination of
revealed a high percentage of them showing multiple drug amoxillin and clavulanic acid, ceftiofur, ampicillin,
resistance to antibiotics like tetracycline, streptomycin, sulfamethoxazole and combination of sulfamethoxazole
ampicillin, sulfamethoxazole, Lincomycin while and trimethoprim respectively. Among the gram negative
susceptibility to antibiotics like ciprofloxacin, Nalidixic bacteria from Owena-Ijesha dam (Fig. 3). It was observed
World Appl. Sci. J., 32 (3): 403-414, 2014
410
Table 7 (Cont'd): Multidrug Resistant bacteria from Owena-Ijesha water samples and their resistance phenotypes (Dam 6)
Sources Bacteria/Strain ID Resistant Phenotypes
Treated water (Gram negatives)
OWIFW Proteus vulgaris (257B) AM, SU, SXT, S, CEF
Treated water (Gram positives)
OWIFW Bacillus pumilus (357B) AM, SU, SXT, E, LIN
OWIFW Bacillus altitudinis (257A) AM, SU, SXT, T, E, RIF, LIN
Municipal Tap 1 (Gram negatives)
OWIM1 Alcaligenes sp (250B2) T, SU, S, K, N, CEF
OWIM1 Acinetobacter junii (364) AM, SU, AMC, CEF
OWIM1 Klebsiella pneumoniae (361) T, AM, SU, S, CEF
OWIM1 Enterobacter sp (360) AM, SU, SXT, AMC
OWIM1 Citrobacter murliniae (363) T, AM, SU, AMC, S, CEF
OWIM1 Citrobacter freundii (362) T, AM, SU, SXT, AMC S, N
OWIM1 Pseudomonas sp (196) T, AM, SU, AMC, S, GEN, K, FF
OWIM1 Alcaligenes faecalis (250A2) T, AM, SU, AMC, S, GEN, K, CEF
Municipal Tap 1 (Gram positives)
OWIM1 Bacillus cereus (250) AM, SU, SXT, LIN
Municipal Tap 2 (Gram negatives)
OWIM2 Brevundimonas naejangsanensis (236B) SXT, S, GEN, K, N
OWIM2 Proteus mirabilis (385B) T, AM, SXT, S, C, N
OWIM2 Chromobacterium sp (366) T, AM, SU, S, GEN, CEF
OWIM2 Alcaligenes faecalis (239) T, AM, SU, SXT, S, K, CEF
OWIM2 Alcaligenes sp (238B) T, AM, SU, S, GEN, K, N, CEF
OWIM2 Pseudomonas sp (244B) T, AM, SU, SXT, AMC, S, GEN, K, C,FF
Gram negative Antibiotics: Ampicillin (AM); Ceftiofur (CEF); Chloramphenicol and Florfenicol (FF); Kanamycin (K), Streptomycin (S) and Gentamycin
(GEN); Tetracycline (T); Nalidixic Acid (N); Sulfamethoxazole (SU); Sufamethoxazole/ Trimethoprim (SXT); Amoxicillin/Clavulanic Acid (AMC)Gram
positive Antibiotics: Sufamethoxazole (SU); Ampicillin (AM); Tetracycline (T); Gentamycin (GEN); Erythromycin; Riframprim (RIF); Lincomycin (LIN);
Ciprofloxacin (CIP), Sulfamethoxazole/Trimethoprin (SXT)OWIFW: Owena-ondo treated water, OWIM1: Owena-ondo municipal 1
Table 7 (Cont'd): Multidrug Resistant bacteria from Owena-Ijesha water samples and their resistance phenotypes (Dam 6)
Sources Bacteria/Strain ID Resistant Phenotypes
Municipal Tap 2 (Gram positives)
OWIM2 Bacillus cereus (370) AM, SU, SXT, LIN
OWIM2 Bacillus sp. (203) AM, SU, SXT, LIN
OWIM2 Lysinibacillus sp (244A) AM, SU, SXT, LIN
OWIM2 Bacillus thuringiensis (238A) AM, SU, SXT, LIN
OWIM2 Bacillus pumilus (244) AM, SU, SXT, E, RIF, LIN
Gram negative Antibiotics: Ampicillin (AM); Ceftiofur (CEF); Chloramphenicol and Florfenicol (FF); Kanamycin (K), Streptomycin (S) and Gentamycin
(GEN); Tetracycline (T); Nalidixic Acid (N); Sulfamethoxazole (SU); Sufamethoxazole/ Trimethoprim (SXT); Amoxicillin/Clavulanic Acid (AMC)Gram
positive Antibiotics: Sufamethoxazole (SU); Ampicillin (AM); Tetracycline (T); Gentamycin (GEN); Erythromycin; Riframprim (RIF); Lincomycin (LIN);
Ciprofloxacin (CIP), Sulfamethoxazole/Trimethoprin (SXT)OWIM2: Owena-Ijesha municipal 2
that out of the total bacteria from each location between and all bacteria isolated from each of the two municipal
0-100%, 75-100%, 0-75%, 48-63%, 50-88%, 88-100%, taps were MDR. Table 6 and 7 showed these MDR
38- 67% showed resistance to tetracycline, streptomycin, bacteria with their various resistance phenotypes. It was
combination of amoxillin and clavulanic acid, ceftiofur, observed that these bacteria include Klebsiella,
ampicillin and sulfamethoxazole and combination of Alcaligenes, Proteus, Pseudomonas, Myroides,
sulfamethoxazole and trimethoprim respectively. From Pseudomonas, Serratia, Providencia, Myroides etc.
Fig. 1 it was observed that 20% of gram negative bacteria Similarity was observed between the MDR bacteria
from the raw water of Owena-Ondo dam were multidrug isolated in our study and that isolated by Bolaji et al. [30]
resistance while the number increased to 88% at each of who also isolated MDR klebsiella, Pseudomonas,
the treated water and first municipal tap respectively. Proteus and Enterobacteria from hospital waste water
From Fig. 3, it was observed that 81% of bacteria isolates from Ede in Southwestern Nigeria. High resistant to
from raw water of Owena-Ijesha were MDR while 50% of antibiotics like streptomycin, ampicillin, tetracycline,
those isolated from the treated water were confirmed MDR Amoxillin/clavulanic acid observed in this study have also
World Appl. Sci. J., 32 (3): 403-414, 2014
411
Fig. 1: Percentage of Gram negative bacteria that were resistant to antibiotics from Owena-ondo water samples
OWODM1= NEPA, Akure OWODM2= Arakale, Akure)
(Rw= 4, Fw= 7, M1= 11, M2= 12, Total no of Bacteria= 34)
Ampicillin (AM); Ceftiofur (CEF); Chloramphenicol (C) and Florfenicol (FF); Kanamycin (K), Streptomycin (S) and
Gentamycin (GEN); Tetracycline (T); Nalidixic Acid (N);, Sulfamethoxazole (SU); Sulfamethoxazole/ Trimethoprim (SXT);
Amoxicillin/Clavulanic Acid (AMC)
Fig. 2: Percentage of Gram positive bacteria that were resistant to antibiotics from Owena-ondo water samples
(OWODM1= NEPA, Akure, OWODM2= Arakale, Akure)
(Rw= 9, Fw= 4, M1= 11, M2= 15, Total no of Bacteria= 39)
Sulfamethoxazole (SU); Ampicillin (AM); Tetracycline (T); Gentamycin (GEN); Erythromycin (E); Rifampin (RIF);
Lincomycin (LIN); Ciprofloxacin (CIP), Sulfamethoxazole/ Trimethoprim (SXT)
World Appl. Sci. J., 32 (3): 403-414, 2014
412
Fig. 3: Percentage of Gram-negative bacteria that were resistant to antibiotics from Owena-Ijesha water samples
(OWIM1= Owena-Igbara oke Tap, OWIM2= Owena-Ijesha Tap).
(Rw= 28, Fw= 2, M1= 11, M2= 3, Total no of Bacteria= 44)
Ampicillin (AM); Ceftiofur (CEF); Chloramphenicol (C) and Florfenicol (FF); Kanamycin (K), Streptomycin (S) and
Gentamycin (GEN); Tetracycline (T); Nalidixic Acid (N); Sulfamethoxazole (SU); Sufamethoxazole/ Trimethoprim (SXT);
Amoxicillin/Clavulanic Acid (AMC)
Fig. 4: Gram-positive bacteria that were resistant to antibiotics from Owena-Ijesha water samples
(OWIM1= Owena-Igbara Oke Tap, OWODM2= Owena-Ijesha Tap)
(Rw= 4, Fw= 3, M1= 4, M2= 6, Total no of Bacteria= 17)
Sufamethoxazole (SU); Ampicillin (AM); Tetracycline (T); Gentamycin (GEN); Erythromycin (E); Rifampin (RIF);
Lincomycin (LIN); Ciprofloxacin (CIP), Sulfamethoxazole/Trimethoprim (SXT)
been reported by Robert. [31]. Most of these antibiotics Among the gram positive bacteria, as shown on
are commonly used or misused as feed additives or Fig. 2 and Fig. 4 showed resistance to ciprofloxacin while
therapeutics agent as reported by Nyamboya et al., [32] high percentage were observed to be resistant to
that reach ground water through their effluents. High lincomycin. At Owena-Ijesha dam (Fig. 4), it was observed
prevalence of MDR Pseudomonas in municipal drinking that 100% of total bacteria from each of the sample
water was also reported by Odjadjare et al. [33]. location were resistant to this antibiotics while between
World Appl. Sci. J., 32 (3): 403-414, 2014
413
75-100% of total bacteria from each of the sample 3. Skipton, S.O., B.I. Dvorak, W. Woldt and S. Wirth,
location of Owena-Ondo dam were resistant to 2007. Drinking water: Bacteria. Neb Guide. A
lincomycin. Among the gram positive bacteria, it was publication of Univesity of Nebraska-Lincoln
observed that the high sensitivity observed to Extension, Institute of Agriculture and Natural
ciprofloxacin and high resistant to lincomycin was Resources.
contrary to what was observed by Chikere et al. [34] 4. Lee, S.W., H.W. Oh, K.H. Lee and T.Y. Ahm, 2009.
among gram positive bacteria from clinical isolates Methylobacterium dankookense sp. nov., isolated
from Nigeria. They observed a high resistant to from drinking water. Journal of Microbiology,
ciprofloxacin but very high sensitivity to gentamycin, 47: 716-720.
lincocin, riframpicin and streptomycin. However, studies 5. Zhang, T., X. Fan, S. Hanadai, Y Kamagata and
have shown that high number of multidrug resistance H.H. Fang, 2006. Bacillus macauensissp. nov., a long-
bacteria in water bodies may be due to misuse of chain bacterium isolated from a drinking water
antibiotics in human health, veterinary medicine, supply. International Journal of Systematic and
aquaculture, agriculture and household products [7, 8]. Evolutionary Microbiology, 56: 349-353.
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surface water and ground water and potentially drinking portable drinking water of Mahabubnagar district
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systems of selected dams in Ondo State, Nigeria is medicine and resultant antimicrobial resistance in
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horizontal gene transfer. The presence of coliform the environment. Journal of Antimicrobial
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not safe and wholesome for human consumption. 10. Barbosa, T. and S. Levy, 2000. The impact of
Therefore, there is the need for government and public antibiotic use on resistance development and
health personal to tackle this problem in order to stop the persistence. Drug Resistant Update, 3: 303-311.
spread. 11. Livermore, D., 2003. Bacterial resistance: Origins,
ACKNOWLEDGEMENTS Disease, 36: 11-23.
We acknowledge Dr Call Douglas of Paul. G. Allen the Examination of Water and Waste Water. Clesceri
School of Global Animal Health, Washington state LS, Greenburg AE and Eation AD (eds). Published
University, USA who allowed the Molecular jointly by American Public Health Association,
Characterization of the bacteria isolates to be carried out American Water Works Association and Water and
in his Lab and also Lisa Lorfe who supplied the technical Environmental Federation. United States of America
knowhow. (Part 9000), 9 of 19-9 of 21.
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... All sampled dams in this study and their various water distribution systems have been described previously [16][17][18]. Two dams were selected for sampling in each of Osun, Oyo and Ondo States. ...
... Previously identified and characterised independent multidrugresistant and florfenicol-resistant bacteria (n = 30) isolated between December 2010 and July 2011 were selected from our former studies [16][17][18] based on their resistance to at least three classes of antibiotics as originally determined by plating samples on agar containing fixed concentrations of antibiotics. Sampling methods and techniques for bacteria isolation have been described previously [18]. ...
... Previously identified and characterised independent multidrugresistant and florfenicol-resistant bacteria (n = 30) isolated between December 2010 and July 2011 were selected from our former studies [16][17][18] based on their resistance to at least three classes of antibiotics as originally determined by plating samples on agar containing fixed concentrations of antibiotics. Sampling methods and techniques for bacteria isolation have been described previously [18]. Selected isolates were re-streaked onto Luria-Bertani (LB) agar supplemented with florfenicol (16 mg/mL) to ensure strain purity and to verify resistance to florfenicol. ...
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Objectives Use of chloramphenicol or its veterinary analogue florfenicol can selectively favour antibiotic-resistant bacteria. Understanding how resistance is mobilised and disseminated among pathogens is vital in knowing how different bacterial taxa might serve as reservoirs of these genes for pathogenic bacteria. Methods Bacterial isolates (n = 30) were selected on the basis of multidrug resistance and resistance to florfenicol from among 296 bacteria originally isolated from drinking water distribution systems in Southwestern Nigeria. Bacterial identification, minimum inhibitory concentration (MIC) determination for florfenicol, PCR detection of florfenicol resistance genes (floR, fexA and cfx) and sequence analysis were employed to characterise the isolates. Results According to sequence data (16S rDNA, v2–v3 region), 30 strains were selected, includingPseudomonas spp. (43.3%), Serratia spp. (13.3%), Proteus spp. (26.7%), Acinetobacter spp. (13.3%) and Providencia rettgeri (3.3%). MICs ranged between >16 μg/mL and >1024 μg/mL. floR was the only resistance gene detected (11/30; 36.7%). The majority of floR-positive isolates (8/11; 72.7%) were Proteus spp. All floR sequences shared 100% identity and 1–2 synonymous substitutions relative to other published sequences. Conclusions floR-positive strains in this study were originally selected randomly without antibiotics. Finding floR in four genera without selective enrichment is consistent with widespread distribution of this resistance trait in drinking water systems in Nigeria. Further work is needed to determine whether human and veterinary antibiotic use practices in Nigeria are contributing to proliferation of this important antibiotic resistance trait and to determine whether the presence of floR-producing strains is compromising human and animal health.
... We previously reported the isolation of several opportunistic bacteria like Acinetobacter spp, Aeromonas spp, Brevundimonas spp, Morganella sp, Psychrobacter, Pantoae spp etc that phenotypically exhibited resistance to gentamicin, kanamycin and streptomycin in municipal drinking water distribution systems from south Western Nigeria. Studies on the physicochemical parameters and microbial qualities of these water samples were also conducted (22)(23)(24). Therefore, this study aimed at detection of AMEs such as aph (3") c , aph (6)-1d d and ant (3") b from these isolates and determination of plamid profiles. ...
... Sample collection, sample sites, bacteria isolates, molecular characterization and antibiotic resistant profiles: Sampling methods, dam description, antibiotics resistant profiles and molecular characterization used in this study were properly elucidated in our previous studies (22)(23)(24)(25)(26). However, for readers' clarity, six dams from three southwestern Nigeria states (i.e. two dams per state) that include Osun, Oyo and Ondo State were selected for this study. ...
... Selected bacteria, AME positive strains and plasmid profiles: Bacteria (n=181) used in this study were selected from our previous study (22)(23)(24). It should be noted that results of the physicochemical parameters and microbial qualities of these water samples had also been reported in these previous studies (22)(23)(24). ...
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Background: Multi-drug Resistant (MDR) bacteria could lead to treatment failure of infectious diseases and could be transferred by non-potable water. Few studies have investigated occurrence of Antibiotic Resistance Genes (ARGs) among bacteria including Aminoglycoside Modifying Genes (AMGs) from Drinking Water Distribution Systems (DWDS) in Nigeria. Here, we aimed at characterization of AMGs from DWDS from selected states in southwestern Nigeria. Methods: One hundred and eighty one (181) MDR bacteria that had been previously characterized using 16S rDNA and showed resistance to at least one aminoglycoside antibiotic were selected from treated and untreated six water distribution systems in southwestern Nigeria. MDR bacteria were PCR genotyped for three AMGs:aph (3″)c, ant (3″)b and aph(6)-1dd. Results: Out of 181 MDR bacteria genotyped, 69(38.12%) tested positive for at least one of the genotyped AMGs. Highest (50, 27.62%) detected gene was ant (3″)c followed by aph (3″)c(33, 18.23%). Combination of aph(3″)c and ant (3″)b in a single bacteria was observed as the highest (14, 7.73%) among the detected gene combination. Alcaligenes sp showed the highest (10/20) occurrence of ant (3″)b while aph(3″)c was the highest detected among Proteus sp (11/22). Other bacteria that showed the presence of AMGs include: Acinetobacter, Aeromonas, Bordetella, Brevundimonas, Chromobacterium, Klebsiella, Leucobacter, Morganella, Pantoae, Proteus, Providencia, Psychrobacter and Serratia. Conclusions: High occurrence of ant (3″)c and aph (3″)c among these bacteria call for urgent attention among public health workers, because these genes can be easily disseminated to consumers of these water samples if present on mobile genetic elements like plasmids, integrons and transposons.
... Safe drinking water is mostly viewed in terms of organic and inorganic contaminants, but also in terms of biological contamination. In this respect, less attention has been given to the role that water may play in the dissemination of antibiotic resistance traits in populations that are exposed to substandard water on a daily basis [2][3][4][5]. ...
... Previous reports indicated that multidrug resistant bacteria are present in drinking water distribution systems from southwestern Nigeria [3][4][5]. These bacteria encoded resistance to a diversity of beta-lactams including ceftiofur, ampicillin, and combination of amoxicillin and amoxicillin/ clavulanic acid. ...
... The aim of this study was to genotype MDR bacteria isolated from our previous studies [3][4][5] for the presence of selected beta-lactamase resistance genes using PCR. ...
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Extended Spectrum Beta-Lactamases (ESBL) provide high level resistance to beta-lactam antibiotics among bacteria. In this study, previously described multidrug resistant bacteria from raw, treated, and municipal taps of DWDS from selected dams in southwestern Nigeria were assessed for the presence of ESBL resistance genes which include í µí±í µí±™í µí±Ž TEM , í µí±í µí±™í µí±Ž SHV , and í µí±í µí±™í µí±Ž CTX by PCR amplification. A total of 164 bacteria spread across treated (33), raw (66), and municipal taps (68), belonging to í µí»¼-Proteobacteria, í µí»½-Proteobacteria, í µí»¾-Proteobacteria, Flavobacteriia, Bacilli, and Actinobacteria group, were selected for this study. Among these bacteria, the most commonly observed resistance was for ampicillin and amoxicillin/clavulanic acid (61 isolates). Sixty-one isolates carried at least one of the targeted ESBL genes with í µí±í µí±™í µí±Ž TEM being the most abundant (50/61) and í µí±í µí±™í µí±Ž CTX being detected least (3/61). Klebsiella was the most frequently identified genus (18.03%) to harbour ESBL gene followed by Proteus (14.75%). Moreover, combinations of two ESBL genes, í µí±í µí±™í µí±Ž SHV + í µí±í µí±™í µí±Ž TEM or í µí±í µí±™í µí±Ž CTX + í µí±í µí±™í µí±Ž TEM , were observed in 11 and 1 isolate, respectively. In conclusion, classic í µí±í µí±™í µí±Ž TEM ESBL gene was present in multiple bacterial strains that were isolated from DWDS sources in Nigeria. These environments may serve as foci exchange of genetic traits in a diversity of Gram-negative bacteria.
... dam 3 in Asejire and dam 4 in Eleyele), and Ondo State (i.e. dam 5 in Owenna Ondo and dam 6 in Owena Idanre) as described in our previous studies [14][15][16] . The sampled dams have been described in detail in our previous works [14][15][16] . ...
... dam 5 in Owenna Ondo and dam 6 in Owena Idanre) as described in our previous studies [14][15][16] . The sampled dams have been described in detail in our previous works [14][15][16] . Bacteria were isolated from 1-ml samples of water by serial dilution and plated on nutrient agar and eosin methylene blue agar. ...
... In this study, 168 MDR bacteria were isolated from raw (n = 65), treated (n = 35), and municipal (n = 68) tap water collected from 6 selected water distribution systems of dams in southwestern Nigeria as described in our previous studies [14][15][16] . Our results showed that MDR bacteria belonging to various genera can be isolated from sampled points. ...
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Background: The emergence of antibiotic resistance among pathogenic bacteria in clinical and environmental settings is a global problem. Many antibiotic resistance genes are located on mobile genetic elements such as plasmids and integrons, enabling their transfer among a variety of bacterial species. Water distribution systems may be reservoirs for the spread of antibiotic resistance. Materials and Methods: Bacteria isolated from raw, treated, and municipal tap water samples from selected water distribution systems in south-western Nigeria were investigated using the point inoculation method with seeded antibiotics, PCR amplification, and sequencing for the determination of bacterial resistance profiles and class 1/2 integrase genes and gene cassettes, respectively. Results:sul1,sul2, and sul3 were detected in 21.6, 27.8, and 0% of the isolates, respectively (n = 162). Class 1 and class 2 integrons were detected in 21.42 and 3.6% of the isolates, respectively (n = 168). Genes encoding resistance to aminoglycosides (aadA2, aadA1, and aadB), trimethoprim (dfrA15, dfr7, and dfrA1), and sulfonamide (sul1) were detected among bacteria with class 1 integrons, while genes that encodes resistance to strepthothricin (sat2) and trimethoprim (dfrA15) were detected among bacteria with class 2 integrons. Conclusions: Bacteria from these water samples are a potential reservoir of multidrug-resistant traits including sul genes and mobile resistance elements, i.e. the integrase gene.
... The Owena-Ijesha dam (1.7 km 2 ) is located near Ilesha, in Osun State. More details about each of these sample sites can be found elsewhere [27][28][29]. ...
... Details on the physicochemical, microbial quality and antibiogram based on sample location of each of these water distribution systems have been reported elsewhere [27][28][29]. Herein we report findings for a total of 105 multi-drug resistant (MDR) bacteria that included strains with resistance to tetracycline that were selected for genotyping. Eighty-two (80.95 %) and 23 (19.04 %) of isolates were Gram-negative and Gram-positive, respectively. ...
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... The description of these dams' water distribution systems selected for this study is in our previous publications [9][10][11]. However, for clarity of this paper, raw water samples were taken from the untreated water from the dam at the point of entering the water treatment plants before passing through the plant. ...
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We describe a new molecular approach to analyzing the genetic diversity of complex microbial populations. This technique is based on the separation of polymerase chain reaction-amplified fragments of genes coding for 16S rRNA, all the same length, by denaturing gradient gel electrophoresis (DGGE). DGGE analysis of different microbial communities demonstrated the presence of up to 10 distinguishable bands in the separation pattern, which were most likely derived from as many different species constituting these populations, and thereby generated a DGGE profile of the populations. We showed that it is possible to identify constituents which represent only 1% of the total population. With an oligonucleotide probe specific for the V3 region of 16S rRNA of sulfate-reducing bacteria, particular DNA fragments from some of the microbial populations could be identified by hybridization analysis. Analysis of the genomic DNA from a bacterial biofilm grown under aerobic conditions suggests that sulfate-reducing bacteria, despite their anaerobicity, were present in this environment. The results we obtained demonstrate that this technique will contribute to our understanding of the genetic diversity of uncharacterized microbial populations.
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"Survival of the fittest " holds good for men and animals as also for bacteria. A majority of bacteria in nature are nonpathogenic, a large number of them, live as commensals on our body leading a symbiotic existence. A limited population of bacteria which has became pathogenic was also sensitive to antibiotics to begin with. It is the man made antibiotic pressure, which has led to the emergence and spread of resistant genes amongst bacteria. Despite the availability of a large arsenal of antibiotics, the ability of bacteria to become resistant to antibacterial agents is amazing. This is more evident in the hospital settings where the antibiotic usage is maximum. The use of antibiotics is widespread in clinical medicine, agriculture, aquaculture, veterinary practice, poultry and even in household products. The major reason for this is the inappropriate use of antibiotics due to a lack of uniform policy and disregard to hospital infection control practices. The antibiotic cover provided by newer antibio
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