Continental J. Microbiology 2: 1 - 4, 2008.

© Wilolud Online Journals, 2008.

BACTERIOLOGICAL ANALYSIS OF CASES DIAGNOSED AS ENTERIC FEVER IN SOKOTO –

NIGERIA

1
M. D. Salihu, 2 S. I. Oboegbulem, 1 O. P. Ajagbonna, 3S. B. Manga, 1A. U. Junaidu and 1M. L. Gulumbe
1
Faculty of Veterinary Medicine, Usmanu Danfodiyo University, Sokoto, 2 Faculty of Veterinary Medicine,
University of Nigeria, Nsukka, 3 Faculty of Science, Usmanu Danfodiyo University, Sokoto.

ABSTRACT
The study was conducted to determine the proportion of confirmed enteric or typhoid
cases as well as non-typhoid salmonellosis among patients clinically diagnosed or
suspected to have typhoid or enteric fever. Serum and fecal samples were collected
from two hundred (200) patients presenting with enteric fever in Sokoto metropolis to
determine the prevalence of non-typhoid salmonella and the reliability of the Widal
test. The serum samples were tested for Widal reaction using Antec Widal Diagnostic
Kit, while the fecal samples from the same patients were subjected to cultural
isolation and identification of salmonellae. The results showed that 132(66%) of the
patients were positive for Widal test. Salmonellae were isolated from 11 (5.5%)
comprising 5(3.8%) and 6(8.8%) from Widal positive and Widal negative samples
respectively. Two of the isolates were non-typhoidal Salmonella, one each from
Widal positive and Widal negative patients. Typhoid Salmonella were recovered from
9(4.5%)of the patients, of which 4(44.4%) and 5(55.6%) were from Widal positive
and Widal negative patients respectively. Other organisms isolated and identified
were Shigella spp., Protozoan cysts and helminth ova.

KEYWORDS: Enteric fever, Widal test; Salmonella, non-typhoid.

INTRODUCTION
Enteric or typhoid fever is endemic and sometimes occurs in epidemic proportions in many tropical
countries including Nigeria (Olubuyide, 1992). The clinical presentation of typhoid fever, usually as
pyrexia of unknown origin is often misleading in a region where malaria is endemic, accounting for about
75% of cases of fever (Grange, 1994). This has lead to under-diagnosis of typhoid fever in many instances.
On the other hand, the practice of making a diagnosis of typhoid fever on the laboratory value of Widal
test, regardless of standard quality control measures, is now quite rampant and responsible for the current
rate of over diagnosis of the disease (Cook, 1985). The antigens that stimulate the antibodies, which are
detected by Widal test, are shared by a very large number of organisms that make up the salmonella genus
as well as by some other related organisms (Guerrant, 1986).

Definite diagnosis of typhoid fever is based on cultures of blood, bone marrow and stool (Homik, 1985;
Edelman and Levine, 1986). In Nigeria, because of the technical difficulties, and cost involved in cultural
isolation and identification of the causative organisms, there is over reliance on the Widal test. The
implication therefore is that if a patient has a “significant typhoidal antibody titre”, treatment of typhoid
fever is often commenced even without a consideration of the clinical signs and symptoms. Patients with
abdominal catastrophes other than typhoid fever but with a “significant Widal test” are labeled as cases of
typhoid fever (Osime and Eghafona, 2003) and are so treated. Many of such patients wrongly diagnosed
and so treated run the risk of having life threatening complications resulting from various pathological
intra-abdominal conditions.

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 M. D. Salihu et al: Continental J. Microbiology 2: 1 - 4, 2008

The present study was conducted to determine the relative proportion of confirmed enteric or typhoid cases
as well as non-typhoid salmonellosis among patients clinically diagnosed or suspected as typhoid or enteric
fever in Sokoto, Nigeria.

MATERIALS AND METHODS

Microbiological Examination of Stool Samples
A total of 200 stool samples from patients presenting as enteric fever cases, were examined over a period of
4 months (February 2004 to May 2004).

Stool samples obtained from patients were examined for salmonellae; by enrichment in selemite F broth,
plating on desoxycholate citrate and brilliant green selective agar, testing for Urease, oxidase and indole
production, and reaction in triple sugar iron agar, lactose, sucrose, manitol, Dulcitol and lysine
(Oboegbulem, 1993; Cowan and Steel, 1993).

Serotyping of the Salmonella isolates was based on Kaufmanns-white scheme (Rowe and Hall, 1980) using
commercial O, H and Vi, antisera from central Research Institute, Kasauli, India. The fecal samples were
also examined for protozoan cysts helminth ova, and hookworm’s larvae.

Widal Agglutination test

Serological diagnosis of enteric fever was carried out by the Widal agglutination technique described by
Gilles and Dodd, (1976) and using commercial Widal test kit obtained from Antec Diagnostic products,
UK. Each serum sample was reacted against the suspension of standard O and H typhoid and paratyphoid
antigens. Agglutination at a titre of 1:80 and above for typhoid or paratyphoid was taken as indicative of
enteric fever in this study.

RESULTS
Incidence of enteric fever (typhoid and paratyphoid)
Enteric fever bacilli (S. typhi and S. paratyphi) were recovered in 9(4.5%) out of 200 patients examined.
Of the 200 patients investigated 132 (66%) gave a significant, titre of 1:80 and above for S. typhi and S.
paratyphi. Positive agglutination lower than 1:80 was not considered significant. The number of patients
showing both significant titres of Widal test and positive stool cultures for enteric bacilli was 4. The
typhoid bacilli were not isolated from stool samples of 128 seropositive patients, but were recovered from 5
seronegative patients. One of the 132 serologically positive patients indicated paratyphoid infection due to
S. paratyphi C, and the 68 seronegative patients also indicated paratyphoid infection with 1 due to S.
paratyphi C.

Non-typhoid Salmonella Infection

Non-typhoid Salmonellae were recovered from two (2) patients; one of the patients was seropositive while
the other was seronegative. The isolates were S. typhimurium and S. enteritidis. The non-typhoid
salmonella accounted for 18.2% of the total salmonella isolates.

Infection due to other organisms

Shigella spp. was isolated from 5(2.5%) of the patients investigated. Four of the isolates were recovered
from Widal positive patients and only one was from the Widal negative patients. Protozoan cysts, helminth
ova were detected in 21(10.5%) of the patients investigated. Seventeen (8.5%) of these were recovered
from patients positive to Widal test and the remaining 4(2%) were from those patients that are negative to
Widal test.

DISCUSSION
Typhoid (enteric) fever infection appears to continue to be of a great public health hazard in developing
countries. Poor sanitation and unhygienic environment aids in the spread of the infection. Definitive
diagnoses of this illness in certain part of the world continue to be a major problem. The definitive

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 M. D. Salihu et al: Continental J. Microbiology 2: 1 - 4, 2008

diagnosis, which is based on the isolation of the causative agent, is hampered by the economic hardship in
the developing countries (Lateef, et. al., 1996). Most cases of suspected typhoid fever have erroneously
been confirmed based on a single serology (Widal test), frequently with the titres much lower than 1:80 in
the study area.

Based on the positive stool cultures for S. typhi or S. paratyphi, only 9 (4.5%) of the patients were shown to
be confirmed cases of typhoid, out of which, 5(2.5%) were from the seronegative group. Many of the cases
of typhoid may in actual fact be due to non-typhoid salmonellae or other gastro-intestinal infections as
revealed in our study.

The Widal test is affected by several factors that can lead to false negative and false positive results. Our
findings showed that 5 of the positive isolate for typhoid bacilli were from the seronegative patient and
only 4 positive isolates were from seropositive patients. This can be associated with factors such as
previous exposure to salmonella infection (Olubuyide, 1992); self medication with antibiotic before
presentation (Agbonlahor, et. al., 1997); recipients of typhoid vaccine (Gupta, et. al., 1985); presence of
fimbrial antigens in the test suspension (Osime and Eghafona, 2003); possibility of cross-reaction of
malaria sera to S. typhi antigen (Lateef, et. al., 1996) and patients that are carrier (Agbonlahor, et. al.;
1997).

Greater proportions of the isolates were S. typhi and less than 20% of the isolates were Salmonella
paratyphi C. This suggests that greater proportion of the population were exposed to Salmonella typhi than
S. paratyphi. This is in line with the findings of Mohammed, et. al., 1992;Olubuyide, 1992 and Osime and
Eghafona, 2003, who reported that Nigeria is endemic for typhoid and that paratyphoid are less commonly
encountered. S. paratyphi is the prevalent paratyphoid bacilli in the area as revealed by our study. The two
serotypes of non-typhoid salmonella recovered in the study, namely S. enteritidis and S. typhimurium are
among the principal causes of food borne salmonellosis. Their isolation in this region is of epidemiological
significance.

CONCLUSION
Patients presenting, as enteric fever cases should therefore be thoroughly examined and samples collected
for bacteriology and serology, to avoid assumed diagnosis of typhoid fever.

REFERENCES
Alaribe, A.A.A.; Ejezie, G.C. and Ezedinachi E.N.U (1998). The prevalence of Salmonella among malaria
patients in Calabar. J. Med. Lab. Sci., 7:34-41

Agbonlahor, D.E.; Aghahowa, M.O.; Idukpaye, O.; Agbonlahor, F.E.; et. al., (1997). Baseline typhoidal
antibody levels in apparently healthy Nigerians. Nig. Qt. J. Hosp. Med. 7(3): 242-245

Cook, G.C.C (1985). Management of Typhoid fever. Trop. Doc. 15: 154 -159

Cowan, S.T and Steel, K.J. (1993). Manual for the identification of Medical bacteria, 3rd (Edn.). Edited by
G.I. Barrow and R.K.A. Fethar. Cambridge, University Press.

Edelman, R.A. and Levine, M.M. (1986). Summary of International Workshop on typhoid fever. Rev. Inf.
Dis 8: 329 – 249.

Gilles, R.R. and Dodds, T.C (1976): the enteric fever, In: Bacteriology Illustrated, 4th edn. Churchill
Livingstone, Edinburgh.

Grange, A.O. (1994). A review of Typhoid fever in Africa. The Postgrad. Med. J. 1(2): 34 – 37

3
 M. D. Salihu et al: Continental J. Microbiology 2: 1 - 4, 2008

Guerrant, R.L. (1986). Salmonella infections. In Harrison’s principles of Internal Medicine, 11th edn.
McGraw Hill, New York, pp. 592 – 599

Gupta, S.P., Gupta, M.S., Bhardwaj, S. and Chugh, T.D. (1985): Current Clinical Patterns of Typhoid
Fever- A prospective study. J. Trop. Med. Hyg. 88: 177-81.

Homik, R.B. (1985). Selective PHC strategies for control of disease in developing World: Rev. Inf. Dis.
1:535 – 546

Lateef, O., Femi, O. Ronke, O. Gbonyega, A., Mojo, A and Sola, A. (1996). Widal agglutination in Malaria
infection. Med. Rev. 3 (6): 5-6

Mohammed, I.; Chikwem, J.O. and Gashau, W. (1992). Determination of Widal agglutination of the
baseline titre for the diagnosis of typhoid fever in two Nigerian states. Scand. J. Immunol. 36 (11): 153 –
156.

Oboegbulem, S.I. (1993). Comparison of two enrichment and three selective media for isolation of
salmonellae from chicken carcass rinse fluids and sewer swabs. Int. J. Food Microbiol. 18: 167 – 170

Olubuyide, I.O. (1992). Typhoid fever in the tropics. Postgrad.Doc.Afr. 14: 37 –41

Osime, C.O. and Eghafona, N (2003). Typhoid Antibodies in a Healthy population: Diagnostic
significance in Nigeria. The Nig. J. Sc. Res. 4(1): 54 – 60

Rowe, B. and Hall, M.L. (1980). Kaufman-white scheme Division of Enteric pathogens. Public Health
Laboratory, London.

Received for Publication: 15/01/2008

Accepted for Publication: 22/02/2008

Corresponding Author:
M. D. SALIHU
Faculty of Veterinary Medicine, Usmanu Danfodiyo University, Sokoto
E-mail- mdsal70@ yahoo.com

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Continental J. Microbiology 2: 5 - 10, 2008.
© Wilolud Online Journals, 2008.

PHYTOCHEMICAL SCREENING AND ANTIBACTERIAL PROPERTIES OF LEAVES OF

PONGAMIA PINNATA LINN. (FABACEAE) FROM INDIA.

S.B.Dahikar1, S.R.Arote1 And P.G. Yeole2

1
Department of Pharmaceutical Microbiology, Sanjivani College of Pharmaceutical Education and
Research, Kopargaon-423603, (INDIA), 2 Institute of Pharmaceutical Education and Research Borgaon
(Meghe) Wardha- 442001(INDIA),

ABSTRACT
Herbal medicine is the most ancient form of health care known to man and are being
used increasingly revealed as dietary supplement to fight or prevent common disease.
Phytochemical screening of the plant leaves reveals the presence of carbohydrates,
alkaloids, flavonoids glycosides, steroids, tannins and saponins. Petroleum ether
extract, Chloroform extract, Ethyl acetate extract and Methanol extracts of Leaves of
Pongamia pinnata Linn. were prepared and antibacterial activity were studied by disc
diffusion method against certain enteric bacterial pathogens such as Escherichia coli,
Staphylococcus aureus, Klebsiella pneumonia, Bacillus subtilis, Enterobacter
aerogenes, Pseudomonas aeruginosa, Salmonella typhimurium, Salmonella typhi,
Staphylococcus epidermidis and Proteus vulgaris. The Methanol extracts had wide
range of antibacterial activity on these bacterial pathogens than the petroleum ether
extract. Ethyl acetate extract were slightly higher antibacterial activity against
bacterial pathogens than chloroform extract. Antibacterial activity of various extract
of leaves of pongamia pinnata was carried in attempt to support the use by medicinal
practitioner for the treatment of enteric infection.

KEYWORDS: Antibacterial activity, Pongamia pinnata, enteric bacterial pathogens.

INTRODUCTION
India is endowed with a rich wealth of medicinal plants. Herbs have always been principal forms of
medicine in India and presently they are becoming popular throughout developing countries, as people
shrine to stay healthy in face of chronic stress and pollution and to treat illness with medicines that works in
concert with body’s own defense. India recognizes more than 2500 plant species which have medicinal
values, However large flora waiting for investigation for their medicinal properties (Kirtikar and Basu
1995).

Pongamia pinnata (L.), locally known as Karanja, is a mangrove plant belonging to the family Fabaceae. It
is a medium size glabrous tree with a short bole and attaining an eight of round18 m and is habitat in the
littoral regions of South East Asia, Australia and Fiji (Chopra et al., 1986; Simin et al., 2002).
Traditionally, its bark is used in pile; leaves are effective as medicated bath and rheumatic pains; seeds are
used in hypertension, bronchitis, whooping cough, skin diseases and rheumatic arthritis (Ballal, 2005;
Tanaka et al., 1992 and Carcache et al., 2003). In primitive areas of Malaysia and India root extracts are
applied to abscesses; other plant parts, especially crushed seeds and leaves are regarded as having antiseptic
properties (Burkill, 1966; NAS, 1980).

In India, seeds were used for skin ailments. Today the oil is used as a liniment for rheumatism; their juice is
used for colds, coughs, diarrhea, dyspepsia, flatulence, gonorrhea, and leprosy. Roots are used for cleaning
gums, teeth, and ulcers also effective in fistulous sores and gonorrhea (Rastogi and Malhotra, 2001; and
Chauhan et al., 2002). Ayurvedic medicine described the root and bark as alexipharmic, anthelmintic, and
useful in abdominal enlargement, diseases of the eye, skin, and vagina, itch, piles, splenomegaly, tumors,
ulcers, and wounds; the leaves, anthelmintic, digestive, and

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 S.B.Dahikar et al: Continental J. Microbiology 2: 5 - 10, 2008.

laxative, for inflammations, piles and wounds; the fruit and seed for keratitis, piles, urinary discharges, and
diseases of the brain, eye, head, and skin (Brown,1995). Unani use the ash to strengthen the teeth, the seed,
carminative and depurative, for chest complaints, chronic fevers, earache, hydrocele, and lumbago (Ahmed
et al., 2004; Yadav et al., 2004).

Enteric or diarrheal infections are major public health problems in developing countries and contribute to
the death of 3.3 to 6.0 million children annually. Enteric bacteria comprised of Salmonella spp., Shigella
spp., Proteus spp., Klebsiella spp., E. coli, Pseudomonas spp., Vibrio cholerae and Staphylococcus aureus
which are major etiologic agents of sporadic and epidemic diarrhea both in children and adults. WHO,
(1985, 1993) reported that 80% populations rely mainly on traditional therapies, involving the use of plant
extracts or their active constituents (Tambekar et al., 2007).

Today there is wide spread Interest in drugs derived from plants for their potential antibacterial activity.
Efforts are these directed to identify plant product used in the treatment of various disease, which have
broad spectrum antibacterial properties (Pathak et al., 1983; Parekh et al., 2005). Therefore the review
revealed that the leaves of Pongamia pinnata (L.), were used in various metabolic disorder, but far their
antibacterial properties were not demonstrated. Hence attempt was made to find out the Phytochemical
constitutes and antibacterial properties of leaves of Pongamia pinnata (L.), against enteric bacterial
pathogens.

MATERIALS AND METHODS

Plant materials: Fresh plant or plant parts of Pongamia pinnata were collected from local region of
Ahmednagar District in India. The leaves were identified by Mr.P.S.N. Rao, Join Director, Botanical
survey of India, Koregaon road, Pune by comparing morphological features (leaf arrangement,
flower/inflorescence arrangement, fruit and seed morphology etc.). The herbarium of the plant specimen
has been deposited at B.S.I. Pune, the voucher specimen No. being BRD1. Fresh plant material were
washed under running tap water, air dried and then homogenized to fine powder and stored in airtight
bottles.

Preparation of extracts: 1.5 kg of the plant material in each batch was exhaustively extracted by soxhlet
extraction method using Petroleum ether, Chloroform, Ethyl acetate and Methanol. The solvent used in
each batch was recovered under pressure until dry extracts were obtained and then labeled and stored
separately at 4 o C in amber colored airtight bottles.

Phytochemical Screening of Plant materials:-

Table 1: Bacterial cultures used in The presence of saponins, tannins, carbohydrates,
study (IMTECH, Chandigarh, India). alkaloids, flavonoids glycosides, steroids, proteins and
alkaloids, were detected by simple qualitative methods
MTCC
Bacterial Pathogens (Khandelwal, 2001).
Number Bacterial cultures: The standard pathogenic
Proteus vulgaris 426 bacterial cultures were procured from IMTECH,
Staphylococcus epidermidis 435 Chandigarh, India and used in the present study
Staphylococcus aureus 96 (Table 1). The bacteria rejuvenated in Mueller-
Escherichia coli 739 Hinton broth (Hi-media laboratories, Mumbai,
Pseudomonas aeruginosa 424 India) at 370C for 18 hrs and then stocked at 40C
in Mueller-Hinton Agar. Subcultures were
Bacillus subtilis 441
prepared from the stock for bioassay. A loopful of
Klebsiella pneumoniae 109 culture was inoculated in 10 mL of sterile nutrient
Salmonella typhi 733 broth and incubated at 370 C for 3 hrs. Turbidity of
5
Enterobacter aerogenes 111 the culture was standardized to 10 CFU with the
Salmonella typhimurium 98 help of SPC and turbidometer.

6
 Table.2 Phytochemical analysis of Pongamia pinnata leaves.

Phytochemical Results of solvent extract

Constitutes Petroleum Chlorofor Ethyl Methanol
ether m extract acetate extract
extract extract
Alkaloid + + + +
Flavonoids - - - +
Carbohydrates - - - +
Glycosides - - - +
Amino acids - - - -
Saponins - - - +
Proteins - - - -
Steroids + + + +
Tannins - - - +
+ =the presence of constitute, – = the absence of constitutes

Antibacterial Activity using disc diffusion method: - The modified paper disc diffusion (NCCLS, 2000)
was employed to determine the antibacterial activity of solvent extract of leaves of Pongamia pinnata (L.).
For antibacterial properties, 0.1 ml bacterial suspension of 105CFU ml-1 was uniformly spread on Nutrient
Agar plate to form lawn cultures. The Petroleum ether, Chloroform, Ethyl acetate and Methanol extracts
were prepared in their respective solvents in such a manner that ultimate amount (in dry form) in each disc
came to 10mg, 8mg, 6mg, 4mg and 2mg. The blotting paper discs (10mm diameter) were soaked in various
diluted extract, dried in oven at 600C to remove excess of solvent and tested for their antibacterial activity
against bacterial pathogens by disc diffusion technique. After incubation of 24 h at 370C, zone of inhibition
of growth was measured in mm. Ampicillin 10mcg (Hi-Media disc) was used as positive control while
discs soaked in various organic solvents and dried were placed on lawns as negative control.

RESULTS AND DISCUSSION

Herbal medicine represents one of the most important fields of traditional medicine all over the world. To
promote the proper use of herbal medicine and to determine their potential as sources for new drugs, it is
essential to study medicinal plants, which have folklore reputation in a more intensified way.

The photochemical investigation (Table 2) of the various solvent extract of Pongamia pinnata showed the
Petroleum Ether extract to content only alkaloids and steroids which, occurring in higher concentration.
The chloroform extract and ethyl acetate extract content similar photochemical constitutes, alkaloids and
steroids in higher concentration, but did not content any Carbohydrates, Flavonoids, glycosides, saponins.
Methanol extract contented Alkaloids, Steroids, Flavonoids, Glycosides, Saponins and Tannins, but did not
contended any proteins.

According to antibacterial profile shown (Table 3). The maximum inhibition of bacterial growth which the
Petroleum ether extract observed only on, Proteus vulgaris, Staphylococcus epidermidis. Staphylococcus
aureus, and, Enterobacter aerogenes, but mild inhibitory effect on Pseudomonas aeruginosa, Escherichia
coli, Salmonella typhimurium and no inhibitory effect on Klebsiella pneumonia and Salmonella typhi.

Chloroform extract inhibited the growth of Staphylococcus aureus, Proteus vulgaris, Staphylococcus
epidermidis and Salmonella typhimurium but mild or negligible effect on Escherichia coli, Pseudomonas
aeruginosa, Salmonella typhi and Klebsiella pneumonia.

Ethyl acetate extract shows maximum inhibitory effect on Staphylococcus aureus, Proteus vulgaris,
Salmonella typhimurium, but no effect on Klebsiella pneumonia, Salmonella typhi.

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 S.B.Dahikar et al: Continental J. Microbiology 2: 5 - 10, 2008.

Methanol extract shows maximum inhibitory effect on Staphylococcus aureus Proteus vulgaris,
Staphylococcus epidermidis, Escherichia coli, Pseudomonas aeruginosa, Enterobacter aerogenes,
Salmonella typhimurium but mild or negligible inhibitory effect on Salmonella typhi Klebsiella pneumonia.

CONCLUSION
The result of the antibacterial assay show promising evidence for the antibacterial effect of leaves of
Pongamia pinnata. Form the above evidence, it is clear that plant extracts have great potential as
antibacterial compounds against enteric pathogens and that they can be used in the treatment of enteric
infectious. This plant can be used to discover bioactive natural products that may serve as leads for the
development of new pharmaceuticals that address hither to unmet therapeutic needs. It is hoped that this
study would lead to the establishment of some compounds that could be used to formulate new and more
potent antimicrobial drugs of natural origin.
.
ACKNOWLEDGEMENT
We acknowledge the contributions of Dr. D.H. Tambekar, Head. P.G. Department of Microbiology, S. G.
Table 3: Antibacterial activity of Pongamia pinnata leaves extracts against enteric bacterial pathogens
(Zone of inhibition of growth in mm, average of 3 readings)
The Petroleum Chloroform Ethyl acetate Negative
Methanol extract
ether extract extract extract controls

Ampicilin
(10mcg)
Bacterial
10mg /disc

10mg /disc

10mg /disc

10mg /disc

Methanol
4mg /disc
2mg /disc

4mg /disc
2mg /disc

4mg /disc
2mg /disc
8mg /disc
6mg /disc

8mg /disc
6mg /disc

8mg /disc
6mg /disc

8mg /disc
6mg /disc

Acetone
4mg /disc
2mg /disc

Ethanol
Pathogens

DW
P. vulgaris 21 19 17 16 15 17 16 15 14 13 18 17 15 13 12 22 20 18 17 16 - - - - 16
S. epidermidis 22 19 18 16 14 14 13 12 11 - 14 13 12 - - 20 18 17 16 15 - - - - 25
S. aureus 25 23 22 19 17 20 19 17 15 14 15 14 13 12 11 24 22 20 18 17 - - - - 24
E.coli 15 14 13 12 11 13 12 - - - 14 13 12 - 20 18 16 15 14 - - - - 11
P.aeruginosa 14 13 12 - - 13 12 - - - 13 12 11 - - 18 17 15 14 12 - - - - 16
B.subtilis 13 12 11 - - 17 16 15 13 12 15 14 13 12 11 22 20 19 17 16 - - - - 18
K.pneumoniae - - - - - - - - - - 12 11 - - - 15 13 12 11 - - - - - 30
S. typhi - - - - - - - - - - - - - - 12 11 - - - - - - - 18
E.aerogenes 20 18 16 15 14 18 17 16 14 12 18 16 14 13 12 22 20 18 17 15 - - - - 14
S.typhimurium 13 12 - - - 15 14 13 12 11 19 16 14 13 12 22 19 17 16 15 - - - - 19

B. Amaravati University, Amaravati for his cooperation through the manuscript and offering her useful
suggestions.

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Received for Publication: 12/04/2008

Accepted for Publication: 22/06/2008

9
 S.B.Dahikar et al: Continental J. Microbiology 2: 5 - 10, 2008.

Corresponding Author:
Samadhan Bhagwan Dahikar
Department of Pharmaceutical Microbiology, Sanjivani College of Pharmaceutical Education and
Research, Kopargaon- 423603, India.
E-mail: sbdahikar10@gmail.com

10
Continental J. Microbiology 2: 11 - 15, 2008.
© Wilolud Online Journals, 2008.

A COMPARISON OF PRESERVATION METHODS OF TRADITIONALLY PROCESSED

DAWADAWA.

Ohenhen R. E., Imarenezor E.P.K., Iyamu M. I. And Aigbokhan F.I.

Department Of Microbiology, Ambrose Alli University, P. M. B. 14, Ekpoma, Nigeria.

ABSTRACT
The preservation of dawadawa using sun drying and oven drying was compared with
fresh dawadawa for bacterial load, moisture content and nutritional content. Bacillus
subtilis and Staphylococcus sp were isolated from the fresh dawadawa while Bacillus
subtilis was isolated from the sun dried and oven dried dawadawa. In terms of
bacterial load, fresh dawadawa had the highest bacterial load of 10 cfu/ml, followed
by sun dried with 10 cfu/ml and the lowest was oven dried which has 10 cfu/ml.
Comparing the moisture content of dawadawa, the sun dried reduced from 100g to
36.5g while the oven dried reduced from 60g to 21.7g. The nutritional content of
dawadawa was accessed based on its protein and glucose value. Oven dried
dawadawa has the highest value for both contents with 52.94g/l and 45.83mg/dl with
a pH of 6.5 respectively. Fresh dawadawa has protein value of 51.21g/l but with the
lowest glucose value of 41.67mg/dl with a pH of 8.1. Sun dried dawadawa has the
lowest protein value of 51.18g/l, but with a better glucose value of 43.75mg/dl with a
pH of 6.4 than the fresh dawadawa.

KEYWORDS: Fermented locust bean, preservation, seeds, dawadawa, glutamate,

seasoning

LITERATURE REVIEW
Dawadawa is the designated name by the Hausa tribe in Nigeria for fermented locust beans (Parkia
biglobosa). It is a culinary product that can be used to enhance or intensify meatiness in soups, sauces and
other prepared dishes. West /Central Africa savannah region consider dawadawa as the most important
food condiment (Odunfa, 1986). The processing of dawadawa starts with harvesting of the seed pods
containing 10- 18 seeds encased in a yellow, sweet farinaceous endocarp. Seeds are separated from the
endocarp and boiled for about 14 hours, cooled and pounded to remove testa; it is either sun or oven dried.
It is washed vigorously, drained severally to remove seeds which still have testa intact, thereby leaving
behind cotyledons which is place in weaved loose bags e.g. jute cocoa bag and compressed by placing
heavy stones and left to ferment for 72 hours. Dawadawa is then put into a mortar, pound to paste before
moulding into desired shapes, after which, it is air dried for 24 hours (odunfa, 1985).

The use of dawadawa in West Africa cannot be overemphasised, especially in the seasoning and flavouring
of meals such as stew and soup. Dawadawa constituted 1.4% of the daily calorie intake and 5% of the total
protein intake (Odunfa, 1985). The flavouring properties of dawadawa are most likely to be due to its
amino acid content in particular glutamate, which contributes to flavour enhancement as well as peptides
and aroma volatile constituents. Volatiles may evolve as a result of the effect of heat on amino acid and
fatty acid constituents of dawadawa (Owen et al., 1997).

The preservation of dawadawa in West Africa is done mostly by sun drying or oven drying. Sun drying is
carried out by spreading of dawadawa in an open air space using a minimum temperature of 85oF and the
optimum temperature of 140oF with humidity below 60%. Oven drying combined the factors of heat, low
humidity and air current. It is carried out using a temperature of 140oF with the oven door left propped
open about 2-6inches (Reynolds et al., 1993).

11
 Ohenhen R. E et al: Continental J. Microbiology 2: 11 - 15, 2008.

This research work is aimed at preserving dawadawa by drying: sun and oven drying and comparing the
moisture levels, nutritional contents and the microbial load of the sun dried, oven dried and freshly
prepared dawadawa.

MATERIALS AND METHODS

COLLECTION OF SAMPLES: Freshly prepared dawadawa was purchased from Ekpoma market which
was divided into three portions. One portion was sun dried; the second portion was subjected to oven
drying while third portion represented the freshly prepared sample.

STERILIZATION OF MATERIALS AND MEDIA USED: Using the autoclave, sterilization of materials
and media (Nutrient agar and MacConkey agar) used was carried out at standard method.

PROCESSING OF SAMPLES: Microbial analysis of samples was done by modified procedure of

Wachukwu (2003).

A. SUN DRYING.
i. MOISTURE LEVEL.
One hundred grams (100g) of dawadawa covered with a muslin cloth to prevent dust and
contamination was sun dried. At one hour intervals for eleven hours, the sun dried dawadawa was
removed from sun and weighed to check for moisture level. A constant moisture level was reached
in 2 days. The dawadawa was manually ground and stored in a bottle.

ii. MICROBIAL LOAD.

Serial dilution of samples was done from 100¹to 100¹0 by emulsifying 1g of samples in 9ml of
sterile distilled water, and shaken vigorously. 1ml of the supernatant was pipette into 9mls of
sterile distilled water. This was serially diluted from 100¹to 100¹0. 1ml of diluents was pipette into
10 sterile Petri dishes and sterile nutrient agar was poured, rocked and allowed to gel before
incubating at 37oC for 24 hours.

iii. NUTRITIONAL STATUS.

The protein value which was carried out using 0.02ml of supernatant was pipetted into a test tube
and 1ml of biuret reagent was added. A standard solution of known concentration (60g/l) was used
and the total protein value was calculated thus;

Absorbance of test/Absorbance of standard X Concentration of standard.

For glucose value, the same procedure as in the protein value was used but read photometrically at
530nm and standard solution of known concentration (100µm/dl) was used. The pH of sample and
it’s titrate able acidity were also read.

B. OVEN DRYING
Sixty grams (60g) of dawadawa was subjected to oven drying at 60⁰C. Moisture level was
checked at 1 hour interval for 10hours and constant moisture was reached on the second day.
Serial dilution method was used to investigate the microbial load. 1 gram was weighed and
dissolved in 100ml of sterile distilled water and same procedure as in sun dried sample was carried
out for the protein and glucose level. The pH and titrate able acidity were also investigated.

C. FRESH DAWADAWA.
One gram (1g) of fresh dawadawa was weighed and serial dilution was also carried out. The
protein and glucose level were observed and pH was also investigated and titrateable acidity.

IDENTIFICATION OF MICROORGANISMS ISOLATED: Standard identification of bacteria was

done by observing their cultural characteristics, gram staining and biochemical characterististics.

12
 Ohenhen R. E et al: Continental J. Microbiology 2: 11 - 15, 2008.

RESULTS
The isolated organisms from fresh dawadawa are Bacillus subtilis and Staphylococcus sp.Bacillus subtilis
was isolated from the oven-dried and the sun dried dawadawa. 100cfu/ml was recorded as contaminant
level for fresh dawadawa while after drying on the second day fell to 100cfu/ml for the sun dried and
100cfu/ml for the oven dried. The result of the moisture levels for the sun dried and oven dried dawadawa,
cultural morphological and biochemical characteristics of bacteria isolated, glucose and protein estimation
of the three samples (fresh, sundried and oven dried dawadawa) is given in tables 1,2 and 3 respectively.

TABLE 1: CHANGES IN THE MOISTURE LEVEL OF SUN DRIED AND OVEN DRIED
DAWADAWA.
TIME ( Hr ) WEIGHT OF SUNDRIED WEIGHT OF OVEN DRIED
DAWADAWA IN GRAMS DAWADAWA IN GRAMS
0 100 60
0-1 80.09 42.60
0-2 60.03 31.60
0-3 48.10 24.80
0-4 47.70 23.10
0-5 40.01 22.60
0-6 38.40 22.70
0-7 37.80 22.10
0-8 37.20 21.70
0-9 36.50 21.70
0-10 36.50 21.70

TABLE 2: CULTURAL MORPHOLOGICAL AND BIOCHEMICAL CHARACTERISTICS OF

BACTERIA ISOLATED.
CULTURAL GRAM REACTION. GROWTH ISOLAT
CHARACTERISTICS. ON MCA. ES.
C C O I G L F S M
A O X N L A R U A
T A I D U C U C L
A G D O C T C R T
L U A L O O T O O
A L S E S S O S S
S A E E E S E E
E S E
E
Creamy large on Gram positive cocci in Pinkish in Staphylo
Nutrient Agar. Smooth clusters. colour ccocus sp
surface, circle and pasty. + - - + A + + + + (lactose
fermenter).
Hide-off white spreading Gram positive Bacilli Bacillus
dull surface. short rods + - + - A - + + + subtilis

Key
MAC = MaConkey Agar.
A = Acid production
+ = Positive.
- = Negative

13
 Ohenhen R. E et al: Continental J. Microbiology 2: 11 - 15, 2008.

TABLE 3: GLUCOSE AND PROTEIN ESTIMATION OF THE THREE SAMPLES (FRESH,

SUNDRIED AND OVENDRIED DAWADAWA).
GLUCOSE VALUE PROTEIN VALUE pH TITRATEABLE
(Mg/dl) (g/l) ACIDITY
SUNDRIED 43.75 51.18 6.4 0.70
DAWADAWA
FRESH DAWADAWA 41.67 51.21 8.1 BASIC
OVENDRIED 45.83 52.94 6.5 2.25
DAWADAWA

DISCUSSION AND CONCLUSION

Bacillus subtilis and Staphylococcus sp were the organisms isolated. These organisms are often been
associated with fermenting food of plant origin and are not pathogenic. (Odunfa 1981). The results obtained
shows that the use of sun drying and oven drying reduced the microbial activity and microbial load of the
organisms involved with dawadawa. Staphylococcus sp was specifically eliminated after drying. On the
first day the initial contaminant level of the fresh dawadawa was 10 cfu/ml. After two days of sun drying
and oven drying, contaminants level fell from 10 cfu/ml to 10 cfu/ml for the sun dried dawadawa and
100cfu/ml for the ovendried dawadawa. This, however, was in conformity with Wachukwu et al (2003).

The use of sun drying and oven drying takes time compared to the use of other preservation methods such
as heat and the use of chemicals. Sun drying also leads to exposure of the food condiment to hazard, such
as entry of organisms present in the air. In order to reduce contamination, Muslin cloth is used to cover the
trays during drying. However when the dawadawa is not properly dried before it is brought under shelter, it
can also reabsorb moisture like in the case of the sun-dried dawadawa. At the fifth hour of drying on the
first day, it weighed 40.01g while on the second day before it was dried, it weight increased from 40.01g to
40.03g. The moisture content of the oven dried dawadawa was also observed to have increased from 22.6g
in the fifth hour on the first day to 23.3g on the second day. This is why proper drying of dawadawa is
required before preservation in order to prevent spoilage and also helps to reduce both microorganism and
enzymes since water is required to be active. Drying also concentrates the soluble ingredients in foods, and
this high concentrates prevents the growth of bacteria, yeasts and moulds. Dried dawadawa will deteriorate
rapidly if allowed to become moist (Reynolds et al. 1993). A constant moisture level was reached at the
eight hour of drying for the oven-dried dawadawa (60g-21.7g) while the constant for the sun dried
dawadawa was reached at ninth hour (100g-36.5g). Comparing the time at which a constant moisture level
was reached for both samples, it can be seen that sun drying is faster than oven drying although the same
amount was not subjected to the different drying methods.

Dawadawa has a great potential as a key source of protein, glucose and also as a flavouring agent. The
protein value for the oven dried dawadawa was highest (52.94g/l), followed by the fresh dawadawa
(51.21g/l) and lowest was the sun dried dawadawa (51.18g/l). The glucose value for oven dried dawadawa
was highest (45.75mg/dl and a pH of 6.5) followed by the sun-dried dawadawa (43.75mg/dl and pH of 6.4)
and lowest was the fresh dawadawa (41.67mg/dl and pH of 8.1). Dawadawa contribute appreciable amount
of protein to the diet of Nigerians. The daily per capital intake of protein from dawadawa is high than from
poultry but less than from beef (Ogunbunmi and Bassir 1980).

Dawadawa has now become a consumer product; it is essential to modernize its production process and
present the product in a better form (packaged) to consumers. The drying of ‘dawadawa’ whether sun
drying or oven drying is desirable and is hereby recommended for commercial purposes as a way of
preservation.

REFERENCES
Odunfa, S.A. (1981). Microorganisms associated with fermentation of African Locust beans during Iru
preparation. Journal of Plants Food. Pp 11, 25.

14
 Ohenhen R. E et al: Continental J. Microbiology 2: 11 - 15, 2008.

Odunfa, S.A. (1985). Biochemical changes in fermenting African Locust beans (Parkia biglobosa) during
iru fermentation. Journal of Food Technology 20: 295-303.

Odunfa, S. A. and Oyewale, O. B. (1986). Identification of Bacillus species from iru, a fermented African
Locust bean product. Journal of Basic Microbiology 26: 101-108.

Odunfa, S.A. (1986). Dawadawa in legume based fermented foods. CRC Press. Boca Raton, Florida. Pp
173-189.

Ogunbunmi, E.M. and Bassir O. (1980). Protein and amino acid content of some African food condiments.
Nutritional Report International Pp. 22, 497.

Owens, J. D., Allagheny, N., Kipping, G.and Ames, J.M., (1997). Formation of volatile compounds during
Bacillus subtlis fermentation of soya beans. Journal of Science and Food Agriculture. 74: 132-141.
Reynolds, S. and Paulitte W. (1993). So easy to preserve. Cooperative extension services. The University
of Georgia. Revised by Judy.

Wachukwu, C. K.,Sokari, T. G.and Wemedu, S.A., (2003). Effect of sun drying and smoking of
Salmonella typhimurium (LT2) during cowpea-dawadawa processing. Journal of Plant Foods for Human
Nutrition. 58(3): 1-7.

Received for Publication: 06/06/2008

Accepted for Publication: 02/07/2008

Corresponding Author:
Ohenhen R. E.,
Department Of Microbiology, Ambrose Alli University, P. M. B. 14, Ekpoma, Nigeria.
Email Addresses: drginaohen@yahoo.co.uk, kimarenezor @yahoo.com.

15
Continental J. Microbiology 2: 16 - 19, 2008.
© Wilolud Online Journals, 2008.

INCIDENCE OF HIV AND HBV INFECTIONS IN PATIENTS ATTENDING UNIVERSITY OF ADO

EKITI TEACHING HOSPITAL
1
Ojo, O.O and 2Daramola, G.O
1
Department of Microbiology, University of Ado-Ekiti, P.M.B 5363 Ado-Ekiti, Ekiti State Nigeria,
2
Laboratory Department, University of Ado Ekiti Teaching Hospital, Ado-Ekiti, Ekiti State, Nigeria

ABSTRACT
Incidence of co-infection of HIV and HBV infections were determined in one
thousand, three hundred and twenty four patients attending the University of Ado-
Ekiti Teaching Hospital between January 2007 and May 2008. Out of this 411 (31%)
were males while 913 (69%) were females. The samples collected from the patients
were screened using DETERMINE and ACONS, HIV and HBV test kits respectively.
A nil incidence was recorded for HIV with HBV. However, 26 (1.96%) of females
and 19 (1.44%) of males were positive for HIV, while 43 (3.25%) of females and 23
(3.25%) of males were positive for HBV. The significance of this finding is
discussed.

KEYWORDS: incidence, HIV, HBV co-infection, patients, teaching hospital

INTRODUCTION
Hepatitis B virus is reputed to be 50-100 times more infectious than HIV (Chessbrough, 2000). It’s been
said that if few drops of HBV-infected blood were dispensed into an Olympic sized standard swimming
pool, the water of the swimming pool will still be infectious. That is despite the fact that such high volume
of water would be far more than enough to dilute to extinction all other known infectious agents.

Acute hepatitis B resembles other forms of viral hepatitis and cannot be distinguished based on history,
physical examination or serum biochemical tests (Beltrami et al, 2000). The diagnosis of acute HBV
infection is confirmed by the demonstration of antibiotics to hepatitis B surface antigen (HBsAg), in the
serum which appears well before the outset of symptoms and before development of antibodies to hepatitis
B core antigen (anti-HBc) and immunoglobulin M (IgM) antibody to HBC, which appear at approximately
the same time as the symptoms (Hoofnagle and Biscelin, 1991, Irwin et al, 1975, Jemson et al, 1987).

The presence of IgM anti-HBc indicates recent HBV infection, usually within the preceding 4-6 month.
The presence of hepatitis B antigen (HBcAg) in serum correlates with HBV replication high litres and
infectivity. Persons who are positive for HBcAg typically have 108 to 109 HBV particles per ml of blood
(Shikata et al, 1977. Tong and Trepo, 1997). In persons who resolve acute HBV infections, antibodies to
HBsAg (anti-HBs) develop and indicate immunity. The persistence of HBsAg for six months after the
diagnosis of acute HBV is indicative of progression to chronic HBV infection (Gordin et al, 1990;
Gerberding 1994, Guptan et al, 1996; Schneiderman and Kaplan 1992, Turner et al, 1989, Donegan et al,
1992)

AIDS, the eventual terminus of unintercepted HIV progression is known to open the door to other
infections, especially the opportunistic ones. The consequences of HIV and HBV co-infection could be
very fatal. Jindal, et al, (2008) determined the prevalence of HIV, hepatitis B and C co-infections in
Amristar, Northern India. Three groups of population at high risk of HIV infections (injecting drug users,
IDUs, truckers and attendees of STD clinic of Amritsar) were studied to determine prevalence of HIV,
hepatitis B and C co-infections. Of the 157 IDUs, 16.6%, 17.8% and 33.7% were found to be positive for
HIV, HBV and HCV respectively. HCV showed significant difference (P < 0.01) and very high rate (8.3%)
of co-infection with HIV. In truckers, they found that maximum seropositivity was associated with HIV
(19%) i-e significantly higher than that of HBV (6%, P < 0.01) and HCV (3%, P < 0.01). In the STD clinic

16
 Ojo, O.O and Daramola, G.O: Continental J. Microbiology 2: 16 - 19, 2008.

attendees, the highest ratio of seroprevalence was that of HIV (4.3%) followed closely by that of HBV
(3.7%) and HCV (2.6%).

This present study was aimed at determining the prevalence of HIV and HBV co-infections among
voluntary and commercial blood donors, patients attending various clinics and those on admission at
University of Ado-Ekiti Teaching Hospital, Ado-Ekiti, Nigeria

MATERIALS AND METHOD

STUDY SITE
The study was carried out at the University of Ado-Ekiti Teaching Hospital, Ado-Ekiti, Ekiti State in south
west Nigeria. The study was carried out between January 2007 and May, 2008.

SAMPLE COLLECTION
Venous blood samples were collected from a total of one thousand three hundred and twenty-four subjects.
The subjects composed of four hundred and eleven males (31%) and nine hundred and thirteen females
(69%) with ages ranging between two day old and 83 years. The subjects were voluntary and commercial
blood donors, in-patients and out-patients of the University of Ado-Ekiti Teaching Hospital.

The sera of the samples were separated out after spinning the samples in the centrifuge for two minutes at
5000rpm.

Retroviral Screening
The samples were screened for the presence or otherwise of antibodies to HIV, using a reliable
immunochromatographic test-kit DETERMINE. This detected the presence of or otherwise of antibodies to
HIV based on the number of colour bands that appeared in the test and control areas at the end of the test.

Hepatitis Screening
Similarly, the samples were screened for the presence or otherwise of antibodies to HBsAg, using
ACONS(R) test-kit, which also utilized the principles of immunochromatography to HBsAg based on the
number of colour-bands that appeared in the test and control areas of the test kit.

RESULT
Forty-five out of the one thousand three hundred and twenty four subjects screened for both HIV and HBV,
were HIV positive, representing on overall HIV seroprevalence of 3.4%.

Table 1: HIV Seroprevalence among the sexes

Female Male Total
HIV (Negative) 887 (66.9%) 392 (29.6%) 1279 (96.6%)
HIV (Positive) 26 (1.96%) 19 (1.44%) 45 (%)
913 (69%) 411 (31%) 1324 (3.4%)

Twenty-six (1.96%) females out of the one thousand three hundred and twenty-four subjects were positive
for HIV, while nineteen (%) males were HIV positive,

Table 2: HBV seroprevalence among the sexes

Female Male Total
Hepatitis B (Negative) 870 (65.9%) 388 (29.3%) 1258 (95%)
Hepatitis B (Positive) 43 (3.25%) 23 (3.25%) 66(4.9%%)
913 411 1324

A total of sixty-six (4.98%) out of one thousand three hundred and twenty four subjects tested positive for
hepatitis B, Out of this figure, fourty-three (3.25%) were females, while twenty three (1.74%) were males.

17
 Ojo, O.O and Daramola, G.O: Continental J. Microbiology 2: 16 - 19, 2008.

The age distribution among the 45 HIV seropositive subjects, shows that six (13.3%) were under 15 years
(these were mostly infants who were victims of vertical HIV transmission from their mothers). Thirty eight
(84.4%) were between 16-60years and one (2.22%) was 62 years old representing 2.22% (see table 3)

Table 3: HIV and HBV Seroprevalence among age-groups

AGE DISEASE
HIV Hepatitis
Under 15 6 (13.3%) 4 (6.1%)
16 – 60 yrs 38 (84.4%) 52 (78.8%)
76 yrs 1 (2.22%) 10 (15.2%)
Total 45 66

The age distribution among the sixty-six hepatitis B positive subjects reveals that four (6.1%) were under
15 years old, fifty-two (78.8%) were aged between 16-60 years and ten (15.2%) were 60 years and above.

DISCUSSION AND CONCLUSION

A most startling revelation of this study is the discovery that the incidence of HIV and HBV co-infection in
Ado-Ekiti is nil. This contrasts sharply with recent similar studies carried out by Pontali and Ferrari (2008)
in Genoa Prison, Italy. They reported an alarming 90% HIV and HCV (hepatitis C) co-infection and also
6.7% HIV/HBV/HCV triple infections among the inmates of Genoa Prison in Italy.

Also this study is in total agreement with that of Jindal et al (2008). They also recorded a nil incidence of
HIV/HBV co-infection in their study carried out in Amristar, India, However, Hoffman and Thio (2007)
think HIV/HBV co-infection is not uncommon.

Why the incidence of HIV/HBV co-infection in Ado-Ekiti is nil is a nagging question that has been thrown
up by this study and this can only be answered by further investigations.

Be that as it may however, this study equally throws up another very vital issue of public concern. This
study has found the overall HIV seroprevalence to be 3.4% whereas the official figure for the state is 1.6%
(FMoH, 2007). While the Federal Government-driven nationwide HIV sentinel survey uses only pregnant
women attending government antenatal clinics (FMoH, 2007), this study used subjects of different ages (2
days to 83 years) from different background and social classes. This shows that the biennially conducted
nationwide sentinel survey is not a true reflection of the real state of HIV seropositivity in Ekiti State and of
course the whole country Relevant federal authorities are advised to re-design the biennial HIV/syphilis
sentinel survey to include subjects of different age groups and social classes and not just pregnant women
attending government antenatal clinics.

REFERENCES
Beltrami EM, Williams I.T, Shapiro CN and Chamberland M.E, (2000). Risk and Management of Blood –
borne infections in Health care Workers. Clin Microbiol Rev. 13,3,385-407

Chessbrough M, (2000). District Laboratory Practice in Tropical Countries, 250 – 251. Cambridge
University Press, UK.

Donegan S.P, Steger K.A, Recla L, Huff R.S, Werner B.G, Rice P.A and Craven D.E, (1992).
Seroprevalence of human immunoffiecieny virus in parturient at Boston City Hospital: Implications for
public health and obstetric practice. Am J. Obstet. Gynaecol, 167, 622 – 629

Federal Ministry of Health (2007). Offincial Report of 2005 National HIV/Syphilis Sentinel Survey, Abuja,
Nigeria.

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 Ojo, O.O and Daramola, G.O: Continental J. Microbiology 2: 16 - 19, 2008.

Gerberding J.L, (1994). Incidence and prevalence of Human immunodeficiency virus, hepatitis B virus,
hepatitis C virus and cytomegalovirus among healthcare personnel at risk for blood exposure: Final report
from a longitudinal study. J infect Dis 170, 1410 – 1417

Gordan F.M, Gilbert C, Hawley H.P and Willoughby A. (1990). Prevalence of human immunodeficiency
virus and hepatitis B virus in unselected hospital admissions. Implications for mandatory testing and
universal precautions. J. Infect Dis 161, 14 – 17

Guptan R.C, Thakur V, Sarin S.K, Benerjee K and Khandekar P, (1996.) Frequency and clinical profile of
precore and surface hepatitis B mutants in Asian – Indian patients with chronic liver disease. Am J
Gastroenlerol 91, 1312 – 1317

Hoffman CJ and Thio CL (2008). Clinical implications of HIV and Hepatitis B co - infection in Asia and
Africa. Lancet Infect Dis Apr 8 (4) 210 – 211.

Hoofnagole JH and Bisceqline Am, (1991), Serological diagnosis of acute and chronic viral hepatitis.
Semin. Liver Dis, 11, 79 – 83

Irwin G.R, Allen AM, Bancroft W.H, Karwack, JJ, Brown H.L, Pinkerton RH, Wilhight M and Top F.H
Jnr, (1975). Hepatitis B antigen in saliva, urine and stool. Infect Immun 11, 142 – 145

Jewson SA, Lemon SM, Baker L.N and Newbold J.E, (1987). Qualitative analysis of hepatitis B. Urine
DNA in saliva and semen of chronically infected homosexual men J. Inject Dis, 156, 299 – 307.

Jindal N, Arora U and Singh K, (2008). Prevalence of human immunodeficiency virus HIV, hepatitis B
virus and hepatitis C virus in three groups of populations at high risk of HIV infection in Amritsar
(Punjab), Northern India. J infect Dis, Jan 61 (1), 79 – 81

Pontali E and Ferrari F, (2008). Prevalence of Hepatitis B virus and/or Hepatitis C Virus co-infection in
prisoners infected with HIV. Int. J Prison Health June 4 (2) 77 – 82.

Schneiderman LJ and Kaplan RM, (1992). Fear of dying and HIV infection versus hepatitis B infection. AM
.J. Infect Control 82, 584 – 586

Shikata T.T, Karasawa T, Abe K, Uzawa T, Suzuki H, Oda T, Imai M, Mayumi M and Montsuyu Y.
(1977). Hepatitis B antigen and infectivity of hepatitis B virus. J. infects Dis, 136, 571 – 576.

Tong S. and Trepo (1997) The HBe-minus mutants of hepatitis B virus. Pp 89-104 In Harrison T.J. and
Zuckerman A.J., The molecular medicine of viral hepatitis. John Willey and Sons, Ltd. Chichester, United
Kingdom.

Turner S.B, Kunches LM, Gordon K.F, Travers PH and Mueller N.E, (1989). Occupational exposure to
human immunodeficiency virus and hepatitis B virus among embalmers: a pilot seroprevalence study. Am J
public Health 79, 1425 – 1426

Received for Publication: 28/06/2008

Accepted for Publication: 24/07/2008

Corresponding Author:
Ojo, O.O
Department of Microbiology, University of Ado-Ekiti, P.M.B 5363 Ado-Ekiti, Ekiti State Nigeria,
Email: walelugba@yahoo.com

19
Continental J. Microbiology 2: 20 - 22, 2008.
© Wilolud Online Journals, 2008.

INCIDENCE OF HIV AND TB CO-INFECTION AMONG PATIENTS ATTENDING THE

UNIVERSITY OF ADO-EKITI TEACHING HOSPITAL
1
Ojo, O.O. and 2Daramola, G.O.
1
Department of Microbiology, University of Ado-Ekiti, Ekiti State, Nigeria.
2
Laboratory Department, University of Ado-Ekiti Teaching Hospital, Ado-Ekiti, Ekiti State, Nigeria.

ABSTRACT
One hundred and nine patients confirmed to be slide-positive to tuberculosis were
selected and screened for presence of antibodies to HIV, to determine the incidence of
HIV/TB co-morbidity and co-infection among patients attending University of Ado-
Ekiti Teaching Hospital. The presence of antibodies to HIV was determined using
immunochromatographic and serial methods. Twenty-nine (26.6%) were found to be
HIV positive. Out of these, ten (9.17%) were males, while nineteen (17.4%) were
females.

KEYWORDS: HIV, TB, co-infection, patients

INTRODUCTION
The magnitude of the morbidity and mortality of Human Immunodeficiency Virus (HIV) and tuberculosis
(TB) co-infection has been put humorously but succinctly, as the marriage between a hundred year old
husband (TB) and a twenty-five years old wife (HIV) which is causing great global havoc. Corbett et al.,
(2003) in their cohort study discovered that all new TB cases in adults (15-49years) were attributable to
HIV infection in an estimated 1.8million death from TB, HIV/AIDS was responsible for 12% of the deaths,
also that TB was responsible for 11% of all adult AIDS death. The rate of progression of TB infection to
TB disease is 30-50 times higher among individuals infected by both TB and HIV as compared to those
infected with TB only (Friedman et al, 1996). It has also been well-established that an HIV-positive
individual who is also co-infected with TB has a 50-60% lifetime risk of developing TB disease as
compared to an HIV negative person who has a 10% risk only (Gordin et. al., 1997). Tuberculosis is known
to be the commonest opportunistic infection and a major cause of mortality among HIV positive person
(WHO, 2004).

Put differently, this means that TB and HIV co-infection is a symbiotic and synergistic association of two
great enemies of mankind, that work hand–in-glove to quickly destroy their victim, in the absence of a
swift intervention. This present study aimed at determining the prevalence of HIV and TB co-infection in
patients attending the University of Ado-Ekiti Teaching Hospital, Ado-Ekiti.

MATERIALS AND METHOD

Study Site
The study was carried out at the University of Ado-Ekiti Teaching Hospital, Ekiti State in South West,
Nigeria, between March and December, 2006.

Sample Collection
Patients who had a history of upwards of three weeks of coughing were directed to the specialist chest
clinic of University of Ado-Ekiti Teaching Hospital, from the out patients Department and other referring
centres within the state. They were asked to produce and submit three sputum samples for three consecutive
days, preferably first thing early in the morning before breakfast.

The blood samples of those who were confirmed to be slide-positive for tuberculosis were later collected
for retroviral screening.

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 Ojo, O.O. and Daramola, G.O: Continental J. Microbiology 2: 20 - 22, 2008.

Screening for tuberculosis

The sputum samples (three for each patient) were processed and screened for the presence of tubercle
bacilli, using the conventional microscopic method.

Retroviral Screening
The sera of the one hundred and nine (109) TB patients were screened for the presence of antibodies to
HIV, using immunochromatographic and serial methods. As in all serial screening methods, all sera that
tested positive for HIV were confirmed to be truly positive with a second different test kit. Out of the 109
TB patients screened for the presence of antibodies to HIV, forty six (42.2%) were males, while sixty-three
(57.8%) were females. They were aged 18 to 85years.

RESULT
Twenty-nine (26.6%) out of the 109 TB patients tested positive for HIV. Ten (9.17%) out of these were
males, while nineteen (17.4%) were females.

Table 1: HIV seroprevalence among the TB patients

HIV POSITIVE HIV NEGATIVE TOTAL
Male 10 (9.17%) 36 (33%) 46 (42.25%)
Female 19 (17.4%) 44 (40.4%) 63 (57.8%)
Total 29 (26.6%) 80 (73.4%) 109

TB patients within the age bracket 18-3years, recorded the highest HIV seroprevalence of 13.8%, followed
by those within the 39-59years bracket, while patients above sixty years of age recorded the lowest HIV
seropositively of 3.67%.

Table 2: Age distribution of HIV-positive TB patients.

AGE HIV POSITIVE HIV NEGATIVE TOTAL
BRACKET
18-38years 15(13.8%) 55(50.5%) 70 (64.2%)
39-59years 10 (9.17%) 19 (17.4%) 29 (26.6%)
60years and 4 (3.67%) 6 (5.5%) 10 (9.17%)
above
Total 29 (26.6%) 80 (73.4%) 109

DISCUSSION AND CONCLUSION

The study reveals an incidence rate of 26.6% co-infection and co-morbidity of HIV/TB among the 109 TB
patients ,as stated in Table1. Thus it is inferable that the TB infection of 26.6% of the study population is
attributable to HIV infection.

This is of course is not surprising as various workers had through numerous studies confirmed the fact that
HIV/AIDS predisposes it victims to not only tuberculosis but other opportunistic infections (DeRiemer e.
al., 2007, Albalak et al., 2007).

The incidence of co-morbidity was found to be highest (13.8%) among subjects aged 18-38years ,as stated
in Table 2, and this is in agreement with the study of Albalak et. al., (2007) who also found a co-morbidity
incidence 13.8% among subjects of comparable age bracket 25-44years.

Furthermore, the female subjects bore the burden of HIV/TB co-morbidity and co-infection more than their
male counterparts with a co-morbidity incidence rate of 17.4% as against 9.17% for males.

And just like it has been pointed out by Albalak et al, (2007), the double jeopardy that HIV/TB co-
morbidity represents can only be reduced through improvements in TB control and treatment and advances
in HIV treatment, prevention as control interventions.

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 Ojo, O.O. and Daramola, G.O: Continental J. Microbiology 2: 20 - 22, 2008.

REFERENCE
Albalak Rachael, O’Brien Richard, Kammerer Steve, O’Brien Sean, Marks Suzanne, Castro Kenneth and
Moore Marisa (2007) Trends in Tuberculosis/Human immunodeficiency virus co-morbidity. Arch Intern
Med. 167: 2443-2452

Corbett L. Elizabeth, Watt Catherine, Walker Neff, Maher Dermot, William Brian, Raviglione Mario, Dye
Christopher (2003). Global trends and Tuberculosis with HIV epidemic. Arch. Intern. Med. 163: 1009-
1021

DeRiemer Kathryn, Kawamura Masae, Hopewell Philip and Daley Charles (2007). Quantitation Impact of
Human Immuradefiency virus infection on Interculosis dynamics Am Thor Soc, 176: 936-944

Friedman L.N, Withan MT, Sing TP and Frienden TR. (1996) Tuberculosis, AIDS and Health among
substance abusers on welfare in New York City. N Eng of Med 334:898-33

Gordin FM, Matt JP, Miller C, Broun LS, Hafner R, John SL (1997). A controlled trial of isoniazid in
persons with anergy and HIV-infection who are at high-risk for pulmonary tuberculosis. N. Engl of Med,
337: 315-20

World Health Organisation (2007). Global tuberculosis, contest, surveillance planning and financing,
WHO Report.

Received for Publication: 23/07/2008

Accepted for Publication: 24/09/2008

Corresponding Author:
Ojo, O.O.
Department of Microbiology, University of Ado-Ekiti, Ekiti State, Nigeria.
Email: walelugba@yahoo.com

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