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    Identification and Antibiogram of Bacteria from Naira Notes Used in Wukari Metropolis, Taraba State, Nigeria Imarenezor Edobor Peter Kenneth 1, * , Olofinlade Olamide Gloria 1 , Joseph Torshian Ugye 2 1 Department of Microbiology, Faculty of Pure and Applied Sciences, Federal University, Wukari, Nigeria 2 Department of Chemical Sciences, Faculty of Pure and Applied Sciences, Federal University, Wukari, Nigeria Email address * Corresponding author Citation Imarenezor Edobor Peter Kenneth, Olofinlade Olamide Gloria, Joseph Torshian Ugye. Identification and Antibiogram of Bacteria from Naira  Notes Used in Wukari Metropolis, Taraba State, Nigeria.  International Journal of Chemical and Biomedical Science.  Vol. 4, No. 3, 2018,  pp. 46-53. Received : July 21, 2017; Accepted : November 8, 2017; Published : May 30, 2018 Abstract: Forty (40) samples of naira notes, of eight (8) different denominations (₦5, ₦10, ₦20, ₦50, ₦100, ₦200, ₦500,  ₦1000) were randomly collec ted from categories of individuals; food vendors, traders, provision stores, cobblers, meat vendors at strategic places in Wukari city for investigation using standard microbiological techniques. The aim of these study was to isolate and identify bacteria allied with the surface of naira notes and there antibiogram. The result showed isolates of  Escherichia coli  40 (19.1%)  , Pseudomonas aeruginosa 34 (16.3%),  Bacillus species 27 (12.9%), Streptococcus  species 17 (8.1%) and  Klebsiella  species 15 (7.7%) and Staphylococcus aureus 67 (31.6%) and Salmonella  species 9 (4.3%). Staphylococcus aureus 67 (31.6%) was more predominant while Salmonella  species 9 (4.3%) was the least. The colony forming units of isolates from the various denomination of ₦5, ₦10, ₦20, ₦50, ₦100, ₦200, ₦500 and ₦1000 were 2.7x10 5 , 3.1x10 5 , 1.8x10 5 , 3.0x10 5 , 3.0x10 5 , 2.9x10 5 , 2.6x10 5  and 1.8x10 5  respectively. The antibiogram of the isolates show that all isolates were susceptible to ciprofloxacin and resistant to augmentin. Other antibiotics (gentamycin, streptomycin, chloramphenicol and ampiclox) has various degree of susceptibility and resistant. In conclusion, naira notes in circulation harbour and serve as vehicles for the transmission of  pathogenic bacteria via human/anthropogenic factors either through poor sanitation and hygiene or poor handling of the naira notes. Contaminated naira notes are potential risks to public health as they can serve as agents for the dispersal of different infectious pathogen which has found to be resistant to most of the commercially marketed antibiotics, making their treatment more difficult. Therefore, care should be taken to prevent these notes coming in contact with pathogenic bacteria. Keywords:  Naira, Bacteria, Antibiogram, Identification, Wukari 1. Introduction The Naira note is the official currency of the federal republic of Nigeria, issued and regulated by the central bank of Nigeria (CBN) which was introduced into the country in 1945 and since then several changes have been made to different denominations used as a legal tender, as a medium for exchange of goods and services, settlements of debts and for deferred payments in economic activities [1]. There are various denominations of naira notes comprising of ₦5, ₦10, ₦20,  ₦50, ₦100, ₦200, ₦500, ₦1000 notes. The naira notes are classified into two categories; the paper category (1000, 500, 200, 100-naira notes) which is made up of 75% cotton and 25% linen and polymer category (50, 20, 10, 5-naira notes) which are made up of polymer such as Biaxially Oriented Polypropylene (BOPP) [2]; [3]. The ₦5, ₦10, ₦20, ₦50,  ₦100, ₦200 -naira notes are the most common and widely used in the daily transactions of Nigerians especially those living in the rural areas, villages and small towns but the ₦500, ₦1000 International Journal of Chemical and Biomedical Science   ; 4(3): 46-53   2018 ISSN: 2472-9574 (Print); ISSN: 2472-9590 (Online)  47 Imarenezor Edobor Peter Kenneth et al. : Identification and Antibiogram of Bacteria from  Naira Notes Used in Wukari Metropolis, Taraba State, Nigeria notes are mostly used by the wealthy populace and in corporate transactions [4]; [5]. The contamination of currency notes could be from several sources; atmosphere during storage, usage and handling [6]. Daily transactions have made currency notes pass through many hands and pathogens have been imposed on them, poor handling practices like spraying during ceremonies where such notes may be trampled upon when they fall to the ground [7]. Paper currencies provide a large surface area as breeding site for pathogens and bacteria are ubiquitous; their ability to contaminate objects such as the naira notes is highly prevalent [8]. Other negative handling attitudes such as the wetting of hands or fingers with saliva or use of contaminated water to lubricate the hands in counting, improper hand washing after using the toilet, coughing and sneezing on hands then exchanging money, and placement and storage of notes on dirty surfaces also leads to contamination and easy transfer of bacterial and cross contamination [9]. Contamination of different objects by potential pathogenic microorganisms is a serious concern of public health because items that pass from one hand to another gives the opportunity of contamination with wide range of pathogenic microorganisms [10]. The physical transfer of material from hands, surfaces and the environment can contaminate paper currencies due to the fact that almost every socio-economic setting regularly handle the paper currencies [11]. Paper currencies acts as a vehicle through which pathogen could be transmitted, most individual do not care how dirty their hands and fingers are when handling money; from the butcher’s  bloody hands, the food vendors wetly oiled hands, the auto-mechanic greasy palms and so many others receive or handle  paper currency with contaminated hands unknowing to them leading to the vast contamination of the notes with microorganisms [12]. There are various sources of microbial contamination on paper currencies, which could be from the counting machine, atmosphere, during storage, usage and  production. Transactions are made with the paper currencies which are transferred from one handler to another, as such  pathogens are imposed on the notes before they are finally deposited in the banks [10]. Archaic handling methods of  placing currency notes underneath brassier with sweat, under the carpet or rugs, in socks and squeezing is still common especially market women, motorcyclists, bus drivers, bus conductors, butchers and meat sellers, are a major source of contamination [7]. Storage of paper currencies in pockets,  polytenes, cotton, leather bags in humid and dark conditions also favour the growth of microorganisms. Moreover, the contamination of the notes can also be traced to dust, soil, water, contact with traumatized skin and saliva when counting the notes [13]. Potentially dangerous bacterial agents that have  been isolated on paper currency include Streptococcus species,  Staphylococcus species  , Escherichia coli, Enterobacter species  , Klebsiella pneumoniae, Salmonella species   [9]. This study is therefore necessary to isolate and identify bacteria from the surface of naira notes and to determine the tolerance and susceptibility of these isolates to different antibiotics. 2. Materials and Methods 2.1. Study Area and Population This study was carried out in the Department of Microbiology, Federal University Wukari, Taraba State,  Nigeria. Wukari metropolis is a large town which is the Headquarter of Wukari Local Government Area of Taraba State. Geographically, Wukari lies between latitude 7°55’42”  North and longitude 9°47’59” East. It has an area of 4,308 km 2  and a population of about thirty thousand (30000). Wukari is home to Federal University Wukari and Kwararafa University. The major languages spoken are Jukun, Kutep, Tiv, Hausa and Fulani [14]. 2.2. Sample Collection A total of forty (40) samples of Nigerian naira notes, which c onsists of eight (8) denominations of naira notes (₦5. ₦10,  ₦20, ₦50, ₦100, ₦200, ₦500, ₦1000) were randomly collected from categories of individuals; food vendors, traders,  provision stores, cobblers, meat vendors at strategic places in the Wukari metropolis. The samples were collected aseptically in a sterile aluminum foil papers from the individuals using disposable hand gloves. The samples were labeled and taken immediately taken to the laboratory for analysis. 2.3. Physical examination of the Currency The currencies were in various physical conditions and were categorized as clean, dirty or mutilated. The term clean describes notes that had a clear appearance without any evident of damage. The term dirty or mutilated describes notes that does not have a clear appearance and are squeezed, damaged, soiled, or held together with cello tapes or paper tapes. 2.4. Preparation of Currency for Analysis The naira notes collected were soaked in 100ml of distilled water for few minutes at room temperature with intermittent shaking to dislodge the cells into the suspension. 2.5. Bacteriological Investigation Standard bacteriological methods were used for the isolation of the various bacteria. The water from the soaked notes was serially diluted in a tenfold serial dilution from 10 -1  to 10 -3  dilution factor and 1ml was inoculated into sterile petri dishes and freshly prepared media of Nutrient agar and MacConkey agar was used. 2.6. Antibiotic Susceptibility Test Standard Antibiotic sensitivity test was carried out using disc diffusion method on nutrient agar. The inhibition zone was measured to ascertain the susceptibility and resistivity of the organism to the antibiotics [15].   International Journal of Chemical and Biomedical Science 2018; 4(3): 46-53 48 3. Result The result of the study shows the presence of various types of bacteria isolates on the naira notes. Although the vast majority of the naira notes used were dirty, mutilated, wrinkled or odorous, the clean looking notes were also found to be contaminated with various kinds of pathogenic bacteria as well. Table 1 shows the physical conditions of the naira notes used of different denominations. Table 2 depicts the overall characteristics of isolated bacterial species from the different sampled naira notes. Table 3 shows the colony forming unit of the denominations of the naira notes. Table 4 shows the frequency of occurrence of bacteria on each of the respective sampled naira notes. Table 5 shows the antimicrobial sensitivity patterns of the different bacterial isolates and table 6 shows the percentage occurrence of bacterial isolates. Figure 1 shows a chart which depicts the frequency of occurrence of  bacterial isolates on the N5, N10 and N20 notes sampled. Figure 2 shows the frequency of occurrence of bacterial isolates on the N50, N100 and N200 notes sampled. Figure 3 shows the frequency of bacterial isolates on the sampled N500 and N1000 notes and Figure 4 shows the percentage of isolation of each bacteria species. Table 1.  Physical conditions of a sample of each naria denominations.   S/No. Denomination Physical condition 1  ₦5 Clean, dirty and mutilated 2  ₦10 Dirty and mutilated 3  ₦20 Clean, dirty 4  ₦50 Dirty, mutilated 5  ₦100 Dirty, mutilated and odorous 6  ₦200 Dirty and mutilated 7  ₦500 Clean, dirty and mutilated 8  ₦1000 Dirty and mutilated Table 2. Overall characteristics of isolated bacteria from naira notes. S/No Morphological Characteristics Gram Staining Biochemical tests Bacterial isolate Coa Cat Cit Oxi Glu Suc Gal Gas H 2 S 1 Pink, round, flat, dry -ve rod - + - - + + + + -  Escherichia coli 2 Milky, round, moist, raised +ve cocci + + + - + - + - - Staphylococcus aureus 3 Greenish, round, flat, dry -ve rod - + + + + - + - -  Pseudomonas aeruginosa 4 Yellowish, round, raised, moist +ve rod - + + - + + - - -  Bacillus sp . 5 Creamy, round, flat -ve rod - + + - + + + + -  Klebsiella sp . 6 Orange, round, raised, moist -ve rod - + - - + - - - + Salmonella sp.   7 Milky, dry, flat, round +ve cocci - - - - + + + - - Streptococcus sp.   Keys: (-) = Negative, (+) = Positive, Coa= Coagulase, Cat= Catalase, Cit= Citrate, Oxi= Oxidase, Glu=Glucose, Suc= Sucrose, Gal= galactose, Gas= gas  production, H 2 S= H 2 S production Table 3. Colony forming unit (CFU/ml) of the denominations of naira notes. S/No Denomination Bacterial Colony Count CFU/ML 1  ₦5 27 2.7 x 10 5  2  ₦10 31 3.1 x 10 5  3  ₦20 18 1.8 x 10 5  4  ₦50 30 3.0 x 10 5  5  ₦100 30 3.0 x 10 5  6  ₦200 29 2.9 x 10 5     7  ₦500 26 2.6 x 10 5  8  ₦1000 18 1.8 x 10 5  Total 209 Table 4.  Frequency of occurrence of bacterial isolates of each denomination of Naira notes.  49 Imarenezor Edobor Peter Kenneth et al. : Identification and Antibiogram of Bacteria from  Naira Notes Used in Wukari Metropolis, Taraba State, Nigeria S/No Denomination E. coli     S. aureus   P. aeruginosa   Bacillus  sp.  Salmonella  sp. Klebsiella  sp.  Streptococcus  sp. Total 1  ₦5 5 10 4 3 1 1 3 27 2  ₦10 7 10 5 4 - 3 2 31 3  ₦20 3 6 2 3 - 2 2 18 4  ₦50 7 8 6 6 1 1 1 30 5  ₦100 4 9 7 2 3 2 3 30 6  ₦200 5 9 5 3 2 3 2 29 7  ₦500 5 9 4 3 1 2 2 26 8  ₦1000 4 5 1 3 1 2 2 18 Total 40 67 34 27 9 15 17 209 Table 5.  Antimicrobial susceptibility test of bacterial isolates against selected antibiotics. S/No. Bacterial Isolate Selected antibiotics Ciprofloxacin Augmentin Gentamycin Streptomycin Chloramphenicol Ampiclox 1  Escherichia coli S R S S R R 2 Staphylococcus aureus S R R S S S 3  Pseudomonas aeruginosa S R R R R R 4 acillus sp.   S R S S S S 5 Salmonella sp . S R S S R R 6  Klebsiella sp   S R S S R R 7 Streptococcus sp . S R R S S S Keys R  –   Resistant S- Susceptible Table 6.  Percentage occurrence of bacterial isolates. S/no Bacterial Isolate Number of colonies (%) occurrence 1  Escherichia coli 40 19.1 2 Staphylococcus aureus 67 31.6 3  Pseudomonas aeruginosa 34 16.3 4 acillus sp.   27 12.9 5 Salmonella sp . 9 4.3 6  Klebsiella sp   15 7.7 7 Streptococcus sp . 17 8.1 TOTAL 209 100   International Journal of Chemical and Biomedical Science 2018; 4(3): 46-53 50 Figure 1.  Frequency of occurrence of bacteria isolates on ₦5, ₦10, ₦20 notes.
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