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African Journal of Biotechnology Vol. 10(9), pp. 1700-1704, 28 February, 2011 Available online at http://www.academicjournals.org/AJB ISSN 1684–5315 2011 Academic Journals Incidence and susceptibility pattern of clinical isolates
from pus producing infection to antibiotics and Carica
papaya seed extract
A. O. Idowu*, B. B. Oluremi and N. I. Seidu
Department of Pharmaceutical Microbiology, Faculty of Pharmacy, Olabisi Onabanjo University, Ago-Iwoye, Ogun State, The prevalence of bacterial pathogens isolated from pus producing infections (wound, eye and ear)
randomly collected from the Medical Microbiology Laboratory of University College Hospital
(UCH),Ibadan, Nigeria was determined. Their susceptibility to selected antibiotics and Carica papaya
seed extracts was investigated. Bacterial pathogen identity was confirmed based on standard methods
which included, Gram stain reaction, colonial morphology on media, lactose fermentation,
catalase,oxidase,coagulase and indole tests. The antibiogram was carried out using standard disc agar
diffusion method employing commercially prepared antibiotic disc (Abtek Limited) of amoxycillin
(25mcg), co-trimoxazole (25 mcg), nitrofurantoin (300 mcg), gentamicin (10 mcg), nalidixic acid (30
mcg),ofloxacin (30 mcg), augmentin (30 mcg) and tetracycline (10 mcg). A total of 58 isolates were
obtained out of which 31 were from wound, 12 from ear and 15 from eye. 41 of the isolates were
recovered from adults, while 17 were from children. Staphylococcus aureus was found to be the most
common organism recovered (51.7%), followed by Klebsiella spp. (11%), Pseudomonas aeruginosa
(11%),Proteus spp. (6.9%) and Escherichia coli (3.4%). From wound swabs, S. aureus isolates was the
highest (51.6%) followed by Klebsiella spp. (22.6%), Pseudomonas aeruginosa and Proteus spp. (9.7%
each) and E. coli (6.45%). In ear infection, P. aeruginosa accounted for 58.3%, S. aureus 25%, while
Klebsiella spp. and Proteus spp. accounted for 16.6% each. In eye infection, S. aureus and Klebsiella
spp. were more frequently isolated with 73.3 and 20%, respectively. The antibiogram studies showed
that all the organisms were highly sensitive to ofloxacin as follows: S. aureus (80%), Klebsiella spp.
(100%), P. aeruginosa (100%), Proteus spp. (100%) and E. coli (100%) while P. aeruginosa (63.6%), S.
aureus (76.6%) and E. coli (100%) were sensitive to gentamicin and Klebsiella spp. (18.2%) and Proteus
spp. (25%) were resistant. The organisms were resistant to all the other antibiotics tested namely:
amoxycillin, co-trimoxazole, nitrofurantoin, nalidixic, augmentin and tetracycline. C. papaya extract
showed antimicrobial activity which compared favourably with the commercial antibiotic discs against
the Gram positive and negative bacteria tested, except Klebsiella spp. The highest activity was
demonstrated against P. aeruginosa.

Key word: Susceptibility, bacterial pathogens, pus, infection, antibiotics, Carica papaya extract.

INTRODUCTION
Infectious diseases are the world’s major threat to human the use of antimicrobial agents is to cure or prevent health and account for almost 50,000 deaths everyday infectious diseases by using the best available agent. (Ahmad and Beg, 2001). The most important reason for The benefits to the individual who deserves treatment must be weighed against the risk of the emergence of resistant microorganisms to the public (Kunin, 1988). Rapid development of multidrug resistance by micro- *Corresponding author. E-mail: [email protected]. Tel:
organisms to available antimicrobial agents has further complicated the threat of infectious diseases to human Table 1. Source and number of isolates used.
Staphylococcus aureus ATCC 25923, Pseudomonas aeruginosa ATCC 27853 and Esherichia coli ATCC 25922 were used as control Children
strains. Diameters of growth inhibition zones were recorded to the nearest mm for an average of 2 readings in each case. Processing of plant materials
C. papaya fruits col ected from Sagamu in Ogun state were authenticated at the Forestry Research Institute of Nigeria (FRIN), Ibadan, Nigeria. The unripe pawpaw fruits were cut open and the health. The choice of antimicrobial chemotherapy is ini- seeds removed and put in clean polythene nylon. The seeds were air dried for two weeks to prevent loss of active compounds. They tial y dependent on clinical diagnosis. However, for many were grounded into powder with electric blender. 500 g of the infections, establishing a clinical diagnosis implies deter- powder was soaked in methanol (96%) for one week and then mining possible microbiological causes which requires filtered. The filtrate was dried to yield a semisolid mass referred to laboratory information from samples col ected, preferably as extract. The crude extract was reconstituted using 50% dime- before antibiotic therapy is begun. Laboratory isolation thylsuphoxide (DMSO) to obtain concentrations of 250, 200, 150, and susceptibility testing of organisms make diagnosis to be established and also make drug selection more
Determination of the activity of C. papaya seed extract on the
The local use of natural plants as primary health reme- bacterial pathogens
dies due to their pharmacological properties is quite common in Asia, Latin America and Africa (Bibitha et al., Antimicrobial assay
2002). Plants have the major advantage of stil being the most effective and cheaper alternative sources of drugs Antimicrobial activity of seed extract of the plant sample was (Pretorian and Watt, 2001). Carica papaya (family evaluated by the cup plate agar diffusion method (Doughari et al., 2007) on Muel er Hinton agar. Aliquots of extract reconstituted in Caricacea) is a monosexual plant of Central American 50% dimethylsulphoxide (DMSO) and diluted to obtain concen- origin. The latex from the leaves has been used as anti- trations of 250, 200, 150, 100 and 50 mg/ml were applied in each helmintics for the treatment of infection of bacterial origin wel in the culture plates previously seeded with the test organisms. A pre-diffusion time of 30 min was al owed at room temperature and This research was undertaken to determine the pre- the plates were incubated at 37°C for 24 h. S. aureus ATCC 25923, valence of bacterial pathogens that are commonly P. aeruginosa ATCC 27853 and E. coli ATCC 25922 were used as control strains. Diameters of growth inhibition zones were recorded responsible for pus producing infections of wound, eye to the nearest mm for an average of 2 readings in each case. and ear and to investigate the susceptibility of the organisms to selected antibiotics as wel as seed extracts Out of the 58 clinical bacterial isolates used in this study, MATERIALS AND METHODS
31 (53.4%) were obtained from wound samples, 12 (20.6%) from ear samples and 15 (25.8%) from eye Collection and preparation of test clinical isolates
samples. 41 (70.6%) of al the isolates were recovered Fifty eight (58) bacterial isolates from wound, eye and ear clinical from adults, while 17 (29.3%) were from children (Table samples were randomly col ected from the Microbiology Laboratory 1). The distribution of the different pathogens showed of University Col ege Hospital (UCH), Ibadan, Nigeria. The isolates that S. aureus was the most prevalent (51.7%), fol owed were obtained on nutrient agar slants and incubated at 37°C for 24 by Klebsiel a spp. (11%), Pseudomonas (11%), Proteus spp. (6.8%) and E. coli (3.4%) (Table 2). From the wound The isolates were re-identified and confirmed using standard microbiological method which included, gram staining, colonial mor- samples, the distribution of the isolates recovered was as phology on media, lactose fermentation, catalase, oxidase, coagu- fol ows: S. aureus (51.6%), Klebsiel a spp. and Proteus spp. (22.6%) each, P. aeruginosa (9.7%), Proteus spp. (9.7%) and E. coli (6.45%). P. aeruginosa was the most common organism isolated from ear specimen (58.3%) Antibiotic sensitivity testing
fol owed by S. aureus (25%), Klebsiel a spp. (8.3%) and Antibiotic susceptibility testing was performed by the disc diffusion Proteus spp. (8.3%). S. aureus isolates was predominant assay on Mul er Hinton Agar as described by Bauer et al. (1966) in the eye samples (51.7%), fol owed by Klebsiel a spp. using the fol owing antibiotics disc: amoxycil in (25 mcg), co- trimoxazole (25 mcg), nitrofurantoin (300 mcg), gentamicin (10 The result of the antibiogram of the pathogenic isolates mcg), nalidixic acid (30 mcg), ofloxacin (30 mcg), augmentin (30 showed that al the organisms were highly susceptible to mcg) and tetracycline (10 mcg). Interpretation of diameter of growth inhibition zone was done by using the manufacturer’s manual. ofloxacin. The proportion of the isolates that were sus- Organisms were scored as either sensitive or resistant. ceptible to ofloxacin were as fol ows: S. aureus (80%), Table 2. Distribution of different pathogens.
Organism
Wound swab (%)
Ear swab (%)
Eye swab (%)

Table 3. Antibiogram profile of bacterial isolates from pus producing infections.
Organism
AMX, Amoxicil in; COT, co-trimoxazole; NIT, nitrofurantoin; GEN, gentamicin; NAL, nalidixic acid; OFL, ofloxacin; AUG, augmentin; TET, tetracycline; Klebsiel a spp. (100%), P. aeruginosa (100%), Proteus et al. (2003) and Bisi et al. (2005) that S. aureus is the spp. (100%) and E. coli (100%) (Table 3).Only three of leading etiological agent in wound infections. However, P. the organisms: S. aureus (76.6%), P.aeruginosa (63.6%) aeruginosa which accounted for only 11% of the total and E. coli (100%) were sensitive to gentamicin, while isolates from the three sites was the predominant Klebsiel a spp. (18.2%) and Proteus spp. (25%) were organism of al the isolates obtained from ear specimen resistant to it. The pathogens were resistant to al the (58.3%). This is in agreement with earlier studies other six antibiotics tested. This showed that these (Adedeji et al., 2007; Ogisi and Osamar, 1982; Selina, categories of the isolates were multiresistant strains 2002) that rated P. aeruginosa as the most common (defined as resistance to at least three classes of bacteria isolated from mild to severe form of external antibiotics). The comparative antibiotics susceptibility of otitis and chronic suppurative otitis media. Pus production bacterial isolates from the different sites of infection is is a common manifestation of infections due to S. aureus shown in Table 4. C. papaya showed antibacterial activity in tissues and sites with lowered host resistance such as against P. aeruginosa, while no activity was recorded damaged skin and mucous membrane,where it may against any of the Klebsiel a spp. Isolates (Table 5). produce skin lesion such as boil or surgical site infections. This possibly explain the high incidence of the organism from bacterial isolates obtained from this pus DISCUSSION
producing infection of eye, ear and wound in this and The knowledge of the bacteriology of an infection and the laboratory susceptibility testing of micro-organism The susceptibility studies showed that the bacterial implicated could make drug selection in antimicrobial pathogen was only highly susceptible to ofloxacin with chemotherapy more rational. Suppurative infection of the varying degree of susceptibility to gentamicin. Resistance skin, ear and eye are common occurrences in to commonly used antibiotics such as amoxycil in, hospitalized and out patients. Wound infection is cotrimoxazole, nitrofurantoin, nalidixic acid and regarded as the most common nosocomial infection in tetracycline was predominant in the organisms.Al the surgical patients (Dionigi et al., 2001). It has been strains of the isolates used were resistant to the associated with increased trauma care, prolonged potentiated B-lactam antibiotic, augmentin. The hospitals stay and treatment. (Bowler et al 2001). phenomenon of multidrug resistance (defined as The result of this study showed that S. aureus which resistance to more than 3 classes of antibiotics) was accounted for 51.7% prevalence is the leading etiological noticed among the isolates in this study. agent of pus producing infections of the eye, ear and There was no defined pattern or difference in terms of wound. S. aureus was the predominant isolates from sensitivity to antibiotic from the isolates obtained from wound (51.6%) and eye (51.7%) specimens. This is in different sites of infection of the eye, ear and wound. The agreement with the reports of Akonai et al. (1991), Shittu emergence of resistance to antimicrobial agents is a Table 4. Comparative analysis of susceptibility pattern of bacterial isolates from pus producing infection in relation to sample
Organism
30 S. aureus(30)
wound
11 Klebsiella spp.
wound
11 P. aeruginosa
wound
4 Proteus spp.
wound
2 E. coli
wound
AMX, Amoxicil in; COT, co-trimoxazole; NIT, nitrofurantoin; GEN, gentamicin; NAL, nalidixic acid; OFL, ofloxacin; AUG, augmentin; TET, global public health problem, especial y among pathogens causing nosocomial infection. (CDC, 1999). This is The demonstration of activity by the extract against the essential y due to improper use of antibiotics by health tested bacterial isolates provide the scientific basis for the professionals, unskil ed practitioners and laypersons, local usage of the plant in the treatment of various pus poor drug quality and inadequate surveil ance programme producing infection such as wound infection and otitis media. It is also an indication that the plant is a potential Adequate epidemiological data on characterization and source for the production of novel drugs for the treatment antibiotic susceptibility of bacterial pathogen is an essential ingredient to guide empiric antibiotic therapy in This study showed that S. aureus is the leading the ambulatory setting. This would reduce or eliminate aetiologic agent in pus producing infections and that errors in empirical selection of either ineffective or ofloxacinand gentamicin may be used in the treatment of expensive drugs, prolonged hospitalization and higher such infections before microbial and sensitivity test are There is need for the evaluation of plants for High level resistance to common antibiotics antibacterial activity because of the twin effect of high encountered in most of the isolates of bacterial cost and the development of resistance to synthetic/ pathogens is an indication that control measures have to conventional drugs used in orthodox practice.In this be put in place, particularly in the administration of study, the seed extracts of C. papaya showed good antibiotics in the hospitals. Patients should be educated antibacterial activity against the Gram negative than the on the consequences of indiscriminate use of antibiotics Gram positive organisms tested. This result is at disparity and there should also be an antibiotic resistance with an earlier study that indicated that plant extracts are more active against Gram positive than Gram negative The high sensitivity of the tested organism to ofloxacin bacteria (Jigna et al., 2006). It however agreed with the may not serve any useful purpose in children where its findings of Doughari et al. (2007), where it was use is contra-indicated. It should also be used with demonstrated that C. papaya extract showed a higher caution in adults because the emerging low level of activity against Gram negative bacteria than Gram resistance may become high in future due to selective Table 5. Comparative antimicrobial activities of C. papaya seed extract with commercial antibiotics
Inhibition zone (mm)
Strain of
organism
C. papaya seed extract (mg/ml)
GEN (mcg)
OFL(mcg)
GEN = Gentamicin; OFL = Ofloxacin; S = Staphylococcus aureus; PS = Pseudomonas aeruginosa; P = Proteus spp; E = Escherichia coli. pressure of exposure as a result of constant use because associated approaches to wound management. Clin. Microbiol. of arbitrary prescription of the antibiotic. Dionigi R, Rovera F, Dionigi G, Imperatori A, Ferrari A, Dionigi P, Dominioni I (2001). Risk factor in surgery. J. Chemother., 13:6-11. Doughari JH, Elmahmood AM, Mansara S (2007).Studies on the REFERENCES
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