Bacteriological assessment of spoilt pharmaceutical products sold in yola metropolis adamawa state, nigeria

International Invention Journal of Medicine and Medical Sciences (ISSN: 2408-7246) Vol. 1(10) pp. 151-156, October, 2014 Available online http://internationalinventjournals.org/journals/IIJMMS Copyright 2014 International Invention Journals Full Length Research Paper Bacteriological Assessment of Spoilt Pharmaceutical
Products Sold in Yola Metropolis Adamawa State,
Ewansiha, J.U*1, Damaris, A.I.1, Ajunwa, O.M1, Umar. A.Y1, Busari, M.B2
1Department of Microbiology School of Pure and Applied Sciences Federal University of Technology, P.M.B 2076 Yola, 2Global Institute for Bioexploration, Federal University of Technology, P.M.B 65 Minna Niger State Nigeria. Forty samples of spoilt pharmaceutical products (oral and ophthalmic) were collected from three
different patent medicine stores and screened for potential bacteria contaminants using standard
laboratory techniques. Of the 40 samples screened, 36 (90%) were seen to be contaminated by 5
different types of bacteria namely Bacillus species, Staphylococcus aureus, Corynebacterium species,
Micrococcus species and Pseudomonas aeruginosa while 4 (10%) were free of contamination. The
range of bacteria count for the oral and ophthalmic products ranges from 1.6 x 102 CFU/ml– 9.9 x 102
CFU/ml and 2.7 x 102 CFU/ml– 9.7 x 102 CFU/ml respectively. The presence of these pathogenic
organisms may contribute to the spoilage of these products and also may be source of infection to the
consumers. Therefore, good manufacturing practices, packaging, storage and distribution should be
enforced.
Keywords: Isolation, Bacteria, Pharmaceutical Products, Spoilt, Oral and Ophthalmic.

INTRODUCTION

Drugs are chemical compounds that may be used on or
of micro-organisms (Willey et al., 2008). The used administered to human or animals as an aid in contaminated products, even where the level of diagnosis, prevention of diseases or other abnormal contamination is low may present potential health conditions for relieve of pain or suffering or to control any hazards to patients (Asaeda et al., 2012). In addition, physiological or pathologic condition (Akerele and such spoilt products constitute wastage and may have Godwin, 2002). Drugs are any chemical substances serious economic implication for the manufacturer. capable of affecting the body. They are used or intended Pharmaceutical products with huge moisture content to modify or explore physiological system or pathological may be contaminated with microorganisms (Villà et al., state for the benefits of the recipient (Gervais et al., 2009). The contaminating microorganisms may spoil the product due to loss of its therapeutic properties. If they Pharmaceuticals of various forms and dosage are are pathogenic, serious infections can arise (Denyer et al., 2004). Modern research identified different types of microorganisms from the raw materials used during (Adeshina et al., 2009). Contaminated pharmaceuticals pharmaceutical productions. These organisms include are considered microbiologically unsafe, if low level of Aspergillus species and Penicillium species (Obuekwe pathogenic or higher level of opportunistic or toxic and Eichie, 2006). A study carried out by Mugoyela and microbial metabolites persist even after death or removal Mwambete 2010, involved structured selection of representative tablets, syrups and capsules from the hospital's outpatient unit. They found the majority of microbial contaminants in non-sterile pharmaceuticals Aspergillus *Corresponding Author Email: [email protected] and Corynebacterium species. Contamination of 152 Int. Inv. J. Med. Med. Sci. pharmaceuticals with microorganisms can also changes samples were inoculated in test tubes containing 9mls of the physical characteristics of the product, including sterile peptone broth. The preparations were incubated breaking of emulsions, thinning of creams, fermentation at 37oC for 24h (Beishir, 1987). Sub-culturing and of syrups, turbidity or deposits, and changes in odor and isolation was done by the use of pour plate method. One color (Gad et al., 2011). The deteriorating effect on the Millilitre (1ml) of enriched samples after incubation was products varies, ranging from introduction of toxic dispensed into sterile petri dishes and 20ml of freshly metabolites and cell fractions to chemical and physical prepared bacteriological media, plated and mixed gently. modifications (Obuekwe et al., 2000). There have been After solidifying, the plates were incubated at 370c for 24 reports of drug-borne human infections worldwide hours. Enumeration of microbial isolates and colony (Mwambete et al., 2009). Villà et al. (2009) reported that counting were done using Gallenkamp digital colony the contamination of thyroid tablets by Salmonella counter and total population expressed as colony muenchen and Pseudomonas aeruginosa, the incidence forming units per ml (CFU/ml). of micro flora in non-sterile preparations generally is influenced by the nature of the ingredients (whether natural or synthetic), the quality of the vehicle, care and Microscopy and Morphological Characterisation
attitude of personnel handling the compounds (Parker, 2000). Gad et al. (2011) recommended stringent limits Pure cultures of bacteria were characterized based on for objectionable microorganisms in drugs intended for colony, microscopic and biochemical tests. The identity use by immune compromised patients as smaller of the isolates was confirmed by comparing their numbers of opportunistic pathogens become infectious microscopic and morphological characteristic with known when resistance mechanisms are impaired, either by taxa (Chessbrough, 2000). immunosuppressive drugs (Sautour et al., 2008). Microbial limits cannot be formulated to cover every Gram Staining
possibility of contamination that may occur. In assessing the results of microbiological testing, the number and The pure isolates were stained according to Gram types of organisms present should be considered in the staining technique (Chessbrough, 2000). The stained context of the proposed use of the product. Thus, in cells were examined under the microscope with oil manufacturing, packaging, storage, and distribution of pharmaceuticals, suitable measures must be taken to microorganisms were characterized by a deep purple or ensure product quality. The aim of this research work violet colour, while the Gram-negative organisms was to isolate and characterize potential spoilage retained reddish pink colour. The shape of the cells was bacteria of pharmaceutical products. also noted (Chessbrough, 2000). MATERIALS AND METHODS
Media used include peptone broth, nutrient agar, Total Colony Counts per Product
chocolate agar and blood agar, the preparation of all media were according to the methods described by The result in Table 1 shows the total colony count by Chessbrough, (2000) product. The highest bacteria count occur in ampliclox A with a total bacteria count of 9.9×102CFU/ml followed closely by gentamycin A (9.7 x 102 CFU/ml) and Collection and Processing of Samples
Gentamycin capsule C (9.2 x 102 CFU/ml) while cotrim had the lowest bacteria count of 1.6×102. Gelucid, Unexpired Pharmaceutical products were collected from ketras, ibuprofen and flagyl had no bacteria growth. different locations in Pharmaceutical stores at Sabogari, Jimeta, and Sangere in Yola, Adamawa State. Products include syrups, tablets and eye drops produced by Range of colony count of bacteria by route of
different manufacturers. All the samples were properly sealed, packaged and transported to the laboratory for microbiological analysis.
The range of colony count of bacteria isolated by route of administration is shown in Table 2. Oral products had 1.6 x 102 – 9.9 x 102 CFU/ml while ophthalmic products Bacteriological Analysis of Samples
had 2.7 x 102 – 9.7 x 102 CFU/ml Distribution of bacteria isolated. Table 3 shows the distribution of bacterial
The bacteriological analysis of samples was carried isolates from the screened pharmaceutical products. out by 10 fold serial dilutions whereby 1ml and/or 1g of Staphylococcus aureus, Bacillus species, Micrococcus Ewansiha et al. 153 Table 1. Total colony count by product
Microbial count
(CFU/ml)
Metronidazole tablet (A) Metronidazole tablet (B) Paracetamol tablet (A) Paracetamol tablet (B) Paracetamol tablet (C) Paracetamol tablet (D) Paracetamol tablet (E) Chlopheniramine tablet( A) Chlopheniramine tablet (B) Chlopheniramine tablet (C) Multivitamin syrup(A) Vitamin A tablet (A) Vitamin B tablet (B) Cotrimoxazole tablet (A) B.Complex tablet (A) B.Complex tablet (B) Gentamycin capsul (A) Gentamycin capsul (B) Cirofloxacin capsul Fluconazole tablet Gentamycin capsule (C) Chloramphenicol capsul (A) Chloramphenicol capsul (B) Benzyline eye drop Ampiclox capsul (A) Ampiclox capsul (B) Ampiclox capsul (C) Ampiclox capsul (D) Cotrimoxazole tablet (B) Lincomycin capsul Analgin tablet (A) Analgin tablet (B) Multivitamin syrup (B) Ketoconazole tablet Magnesium trisilicate Ibuprofen tablet Alphabets in parenthesis represents manufacturer Table 2. Range of colony count of bacteria by route of administration
Microbial Count Range
Specification BP (CFU/ml) British
(CFU/ml)
Pharmacopoeia, (2008)
1.6×102-9.9×102 2.7×102-9.7×102 BP = British Pharmacopoeia 154 Int. Inv. J. Med. Med. Sci.

Table 3. Distribution of bacteria isolated from pharmaceutical products
Products (g/ml)
Bacteria isolate
Total No. Of isolates
(per gram)
Metronidazole tablet (A) Pseudomonas aeruginosa, Staphylococcus aureus Metronidazole tablet (B) Bacillus species, Micrococcus species, Staphylococcus aureus Paracetamol tablet (A) Corynebacterium species, Bacillus species Paracetamol tablet (B) Staphylococcus aureus Paracetamol tablet (C) Micrococcus species, Bacillus species, Staphylococcus aureus Paracetamol tablet (D) Micrococcus species, Bacillus species, Staphylococcus aureus Paracetamol tablet (E) Micrococcus species, Bacillus species, Staphylococcus aureus Chlopheniramine tablet( A) Pseudomonas aeruginosa, Staphylococcus aureus Chlopheniramine tablet( B) Pseudomonas aeruginosa Chlopheniramine tablet( C) Bacillus species, Micrococcus species, Staphylococcus aureus Bacillus species, Corynebacterium species Bacillus species, Staphylococcus aureus Multivitamin syrup(A) Micrococcus species, Bacillus species Vitamin A tablet (A) Bacillus species, Pseudomonas aeruginosa Vitamin A tablet (B) Bacillus species, Staphylococcus aureus Staphylococcus aureus Cotrimoxazole tablet (A) Bacillus species, Micrococcus species Ampiclox capsule (A) Pseudomonas aeruginosa, Staphylococcus aureus Ampiclox capsule (B) Pseudomonas aeruginosa, Micrococcus species Ampiclox capsule (C) Bacillus species, Micrococcus species Ampiclox capsule (D) Bacillus species, Staphylococcus aureus B.Complex tablet (A) Corynebacterium species, Bacillus species B.Complex tablet (B) Corynebacterium species, Bacillus species Benzyline eye drop(A) Pseudomonas aeruginosa, Staphylococcus aureus Benzyline eye drop (B) Pseudomonas aeruginosa, Micrococcus species, Staphylococcus Cirofloxacin capsule Corynebacterium species, Bacillus species Fluconazole tablet Staphylococcus aureus, Bacillus species Benzyline eye drop (C) Micrococcus species, Corynebacterium species, Staphylococcus Chloramphenicol capsule (A) Pseudomonas aeruginosa, Micrococcus species, Staphylococcus Chloramphenicol capsule (B) Staphylococcus aureus, Corynebacterium species Benzyline eye drop (D) Staphylococcus aureus, Micrococcus species Lincomycin capsule Bacillus species, Staphylococcus aureus Cotrimoxazole tablet (B) Bacillus species, Staphylococcus aureus Analgin tablet (A) Pseudomonas aureus, Staphylococcus aureus Analgin tablet (B) Staphylococcus aureus, Micrococcus species Multivitamin syrup (B) Micrococcus species, Corynebacterium species Ketoconazole tablet Magnesium trisilicate Ibuprofen tablet NG = No Growth, Alphabets in parenthesis represents manufacturer species, Corynebacterium species, and Pseudomonas Staphylococcus aureus had the highest prevalence aeruginosa were predominant. (32%) while Corynebacterium species were least Prevalence of bacterial isolated
DISCUSSION
The prevalence of bacterial isolated from the pharmaceutical products is shown in Figure 1. The result of this research showed that the samples Ewansiha et al. 155 Figure 1. Frequency of bacteria occurrence.
tested had satisfactory bacteria levels compared to the practice (GMP) and ensuring compliance through regular British Pharmacopoeia specification of 103 CFU/ml. The monitoring of non-sterile products. The types of organisms isolated were Staphylococcus aureus (32%), organisms isolated in this study suggest the main Bacillus species (27%), Corynebacterium species (10%), sources of contamination of these products were from Micrococcus Pseudomonas the processing unit, handlings, and storage. Bacillus aeruginosa (14%). species which was isolated from the pharmaceutical contaminated as it had 9.9 x 102 CFU/ml. The least products are ubiquitous organisms and considered contaminated sample was Cotrim Cough Syrup with a harmless in this study, though undesirable because of viable count of 1.6 x 10 CFU/ml. The low count recorded their potential to spoil the products. The presence of this in Cotrim Cough Syrup may be attributed to the organism in the products can be traced back to the raw incorporation of trisodium citrate together with sugar materials used in the production of pharmaceuticals content of the syrups which provide high osmotic since this organism is present in the water and un-sterile pressure that is inhibitory to many microorganisms air in the manufacturing environment which need (Muhammed and Umoh, 2009). However the presence fumigation (Kulshrestha et al., 2008). The manufacturing of Staphylococcus aureus reflects its nature of habitation equipment may be handicapped by a number of i.e. human skin which could easily contaminate the designed faults (Kulshrestha et al., 2008). The extent of products during processing. The organisms being a microbial contamination depends on a number of factors normal flora of the body can easily contaminate the products during handling and processing by personnel. microorganisms; oxygen and the factors determining the outcome of medicament-borne infections include the type and degree of microbial contamination, root of concentration, Staphylococcus aureus thrive well in fairly administration and state of the patient's immune system. high concentration of sugar (Madigan and Martinko, Gram positive isolates were mainly Staphylococcus 2006). The high number of Staphylococcus aureus in aureus, Corynebacterium species, Bacillus species and these preparations suggests that they are able to Micrococcus species. Pseudomonas aeruginosa was the tolerate the presence of preservatives in such products only gram negative rod. The presence of bacteria in the as reported by (Cundell, 2009). The type of bacterial unhygienic practices and non-adherence to good contaminations possibly water, personnel, handling and manufacturing environment; unhygienic handling of the the environment (Okunlola et al., 2007). These bacterial products and lack of microbiological in-house control species have previously been associated with drug may also have contributed to the microbial load in some contaminations (Ibezim et al., 2002; Takon and Antai, samples. The strict compliance to this practice will 2006). The isolation of Staphylococcus aureus in these consequently reduce the incidence of contamination and products indicates a possible health risk and the need to guarantee good quality products (Takon and Antai, reduce the degree of contamination of such products by 2006). In addition, some of the products without enforcing official guidelines such as good manufacturing microbial contamination might be as a result of the 156 Int. Inv. J. Med. Med. Sci. presence of preservatives. Madigan MT, Martinko JM (2006). Brock Biology of Microorganisms (11th edition).Pearson. ISBN 0-13-196893-9. Mugoyela V, Mwambete DK (2010). Microbial contamination of nonsterile pharmaceuticals in public hospital settings, Therapeutics REFERENCES
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