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Occurrence of Antibiotic Resistant Bacteria in Environmental Wastes_Nature

Occurrence of Antibiotic Resistant Bacteria in Environmental Wastes_Nature

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Published by Nawa Raj Dhakal
This study describes the antibiotic susceptibility pattern of
the bacteria isolated from sewage sludge, biowaste and bioslurry samples.
This study describes the antibiotic susceptibility pattern of
the bacteria isolated from sewage sludge, biowaste and bioslurry samples.

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Categories:Types, Research, Science
Published by: Nawa Raj Dhakal on Jan 26, 2010
Copyright:Attribution Non-commercial


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R.C. Poudel, D.R. Joshi, N.R. Dhakal and A.B. Karki / Our Nature (2009) 7: 151-157151
Occurrence of Antibiotic Resistant Bacteria inEnvironmental Wastes
R.C. Poudel
, D.R. Joshi
, N.R. Dhakal
and A.B. Karki
 National College, Khusubi, Kathmandu
Central Department of Microbiology, Kirtipur, Kathmandu
Alternative Energy Promotion Centre, Khumaltar, Lalitpur 
Biogas Sector Partnership (BSP), Nepal
Received: 21.01.2009, Accepted: 09.04.2009
Microbial resistance to antibiotics has been emerging in environmental isolates.
This study wascarried out from October 2008 to January 2009 to describe the antibiotic susceptibility pattern of the bacteria isolated from sewage sludge, biowaste and bioslurry samples. A total of 49identified isolates were taken for antibiotic susceptibility test. Amikacin and Gentamicin werethe effective antibiotics for the Gram negative bacteria, comparatively
Escherichia coli
was themost sensitive. Similarly, all isolates of 
Staphylococcus aureus
spp. weresensitive to Tetracycline and Erythromycin respectively and none of these isolates were resistantto Vancomycin. Out of 13 S
. aureus
, five (38.5%) were Methicillin resistant
(MRSA). The multidrug resistant (MDR) isolates accounted for 46.9%, the highestpercentage of MDR isolates was seen in
Pseudomonas aeruginosa
(9/12, 75.0%). The propermanagement of multidrug resistant bacteria present in waste is suggestive for environmental andpublic health.
 Key words
Multidrug resistant bacteria, Bio-waste, Sewage sludge, Bioslurry
Sewage sludge and biowaste contain humanand animal pathogens in high number(Lepeuple
et al.,
2004). The prophylactictreatments, such as medication andsanitation, can help to control the pathogensbut these treatments are increasinglyexpensive and decreasingly effective sincedrug resistance can develop inmicroorganisms (Marchaim, 2007).Antibiotic resistant bacteria are now foundin large numbers in virtually everyecosystem.Resistance against antimicrobials maycause severe problems in human and animalhealthcare (Sahlström, 2006). Thedevelopment of bacterial antimicrobialresistance is neither an unexpected nor anew phenomenon. The consequences of over use of antibiotics in human, animal andagriculture exposes both terrestrial andaquatic environments which accelerate forthe development of drug resistant bacteria inthe environment and also change thecommunity structure/diversity of environmental bacteria (Andersson, 2003;Food Safety, 2008). Due to the applicationof manure from medicated livestock beingapplied to agricultural soils, multiple drug
Our Nature (2009) 7:151-157
R.C. Poudel, D.R. Joshi, N.R. Dhakal and A.B. Karki / Our Nature (2009) 7: 151-157152resistance has developed in the micro-floraand intestinal flora of livestock (Halling-Sorensen
et al.,
1998). Besidesantimicrobial substances, the non-antibioticsubstances that are frequently found insewage such as disinfectants, sterialints, andheavy metals increase selective pressure onmicroorganisms to develop resistance tothese agents (Baquero
et al
., 1998).The presence of large quantities of specific antibiotics in the environment, thedisposal of unused drugs and the use of antibiotics in raising food animals hascontributed significantly to the global poolof antibiotic resistant bacteria amonghuman, animal and the environment andthus create a serious global human andanimal health problem. We designed a studyto describe antibiotic susceptibility patternof the gram positive and gram negativebacterial isolates from waste samples in theenvironment.
Materials and methods
A total of 36 waste samples including asewage sludge, seventeen different bio-waste and eighteen bio-slurry samples wereselected for the isolation of gram positiveand gram negative bacteria. Sewage sludgewas collected from sewerage treatment plantat Guheshwori in Kathmandu whereas bio-waste (fruit and vegetable waste, poultrymanure, animal dung, kitchen waste etc.)and bioslurry (digested residue) sampleswere collected from different institutionaland household biogas plants withinKathmandu valley. All the samples werecollected by random sampling techniqueand were analysed in MicrobiologyLaboratory of National College,Kathmandu. Different microbiologicalmedia were used for the isolation of different bacteria which were identified bytheir cultural, morphological andbiochemical characteristics.Altogether 49 bacterial isolates weresubjected to antibiotic susceptibility testingby using a disc diffusion method inaccordance with Clinical and LaboratoryStandards Institute (CLSI).
producing inhibition less than 10mm zone diameter or producing noinhibition to Oxacillin discs (1µg) wereconsidered resistant to Oxacillin and theorganism was defined as MRSA (MH,2002; Savas
et al
., 2005). On the basis of sensitivity pattern of the isolates, bacteriathat were resistant to
2 classes of antibiotics were considered as multi drugresistant bacteria (Dahal
et al
., 2005).
A total of 49 bacterial isolates wereidentified which belong to five genera andtheir pattern of distribution is shown inFigure 1.Gram negative bacteria exhibitedelevated resistance towards Amoxycillin.All isolates of 
 Escherichia coli
weresensitive to Gentamicin and Amikacin. Themost effective antibiotics for
were found to be Gentamicinand Amikacin as 11(91.67%) isolates weresensitive. Similarly, out of 5
 spp., 4 (80%) were sensitive toCiprofloxacin, Cotrimoxazole,Chloramphenicol and Gentamicin (Table 1).The antibiotic susceptibility test for
Staphylococcus aureus
showed thatVancomycin and Tetracycline was the mosteffective as all 13 (100%) isolates weresensitive. Out of 13 isolates, 5(38.46%)
R.C. Poudel, D.R. Joshi, N.R. Dhakal and A.B. Karki / Our Nature (2009) 7: 151-157153isolates were found to be MRSA. All
spp. were sensitive beVancomycin and Erythromycin and resistantto Cephalexin and Ceftazidime (Table 2).Out of 49 isolates taken for antibioticsusceptibility test, 23 (46.9%) were found tobe MDR isolates. The results are shown inTable 3.
Sewage sludge and biowastes harbor a widerange of pathogenic and indicatororganisms. The composition in pathogensdepends on the type of waste studied. Atotal of 109 bacteria were identified fromsewage sludge, biowaste and bioslurrysamples, which belong to 12 genera. Dudley
et al.
(1980); Strauch (1991); Carrington(2001); Lepeuple (2004) and Sahlström
et al
. (2008) have reported similar genera of bacteria from the sewage sludge andbiowaste.A total of 49 environmental isolates of 5genera were taken for antibioticsusceptibility test against differentantibiotics.
 Escherichia coli
and faecalStreptococci are the indicator organisms andthey were frequently isolated even from thedigested residues (bio-slurry). While otherbacteria such as
spp. and
spp.are pathogenic and most commonlyassociated with humans diseases. Theselection of antibiotics for theirsusceptibility test was dependent upon thetype of antibiotics used for the particulartype of bacteria. The increasing use of antibiotics in humans, animals andagriculture may be the important factor indeveloping the antibiotic resistant bacteria.The antibiotic susceptibility pattern among11
 Escherichia coli
isolates showed that90.91% the isolates were resistant towardsat least one of the antibiotics used and45.46% of the total
isolates were multidrugresistant.
All isolates were sensitive toGentamicin, Amikacin and Ceftazidime andthe higher resistance was seen toAmoxycillin (90.91%). The antibioticresistance patterns obtained by Lin
et al
.(2004) among 113 enteric bacteria isolatedfrom Mhlathuze river showed that 94.7%were resistant to at least one class of antibiotic, 75.2% were multiresistant and allisolates were sensitive to Gentamicin.Similarly, the study conducted by Alhaj
et al
. (2007) found that 61.2% of the totalenvironmental isolates of 
 E. coli
were multiresistant. The lower percentage of multidrugresistant
 E. coli
found in present study maybe affected by the location (Pillai
et al
.,1997).Among 12 environmental
isolates, 75% were found to bethe multidrug resistant while 22.22% wereresistant towards more than 3 antibiotics of different class. The higher susceptibility wasobserved to Amikacin and Gentamicinwhich exhibited 91.67% followed byCiprofloxacin (83.34) and Ceftazidime(66.67%) and all isolates were resistanttowards Tetracycline and Cephalexin.Shahid and Malik (2005) reported that 96%of clinical
Pseudomonas aeruginosa
isolateswas multi drug-resistant and the majority(71.4%) were resistant to 5 or moreantibiotics. The clinical
isolates are more resistant to theantibiotics than environmental isolates(Shahid and Malik, 2005). 40% of theisolates of 
spp. were multidrugresistant (MDR), the higher resistance was

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