CHARACTERIZATION AND SUSCEPTIBILITY OF ESCHERICHIA COLI ISOLATED FROM INFANTS DIARRHOEA TO MOMORDICA CHARANTIA AND AESCHYNOMENE UNIFLORA EXTRACTS
CHARACTERIZATION AND SUSCEPTIBILITY OF ESCHERICHIA COLI ISOLATED FROM INFANTS DIARRHOEA TO MOMORDICA CHARANTIA AND AESCHYNOMENE UNIFLORA EXTRACTS
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Date
2014-10
Authors
Isma’ila, AHMED
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Abstract
A study was conducted to characterize and determine the susceptibility of Escherichia coli isolates obtained from infants‘ diarrhoea stool samples to extracts of Momordica charantia Linn and Aeschynomene uniflora E. May in Kano state, Nigeria. Five hundred and eighty six (586) diarrhoeal samples of infants 0-5 years attending some hospitals in Kano state were examined for the presence of Escherichia coli. Standard methods of culturing, microscopy, Gram staining, biochemical tests and serotyping were used for the identification and characterization of the isolates. Samples were collected from some hospitals from the three senatorial zones. Some demographic data were also obtained from the infants‘ mothers/caregivers. Most common E. coli serotype isolates include enterotoxigenic E. coli O148:H28 and enterohaemorrhagic E. coli O157:H7 with number positive of 48 (23.53%) and 41 (20.10%) respectively and enteropathogenic E. coli O112 and enteroheamorrhagic E. coli O26:H11 with number positive of 35 (17.16%) each. Enteroinvasive E. coli strains O124 and O143 were the least common in the study with the least number positive of 0 (0.00%) and 7 (3.43%) respectively. Prevalence of 34.81% (n=204) of infants‘ diarrhoea was obtained from the study. Males infants‘ were more infected than females with number of males and females positive of 113 (19.28%) and 91 (15.53%) respectively. The prevalence is highest at age range of 24-35 month with total number infected of 54 (9.22%) while least infection was obtained in 0-11 month with number infected of 30 (5.12%). The results of susceptibility pattern of E. coli isolates to antibiotic showed that the E. coli are more susceptible to ciprofloxacin, ceftriaxone, augmentin and ofloxacin with numbers of susceptible isolates of 169 (83%), 159 (78%), 155 (76%) and 153 (75%) respectively. The isolates are however resistant to ampicillin, amoxycillin, sulphamethoxazole/trimethoprim and tetracycline with number resistance of 182 (89%), 170 (83%), 105 (51.47%) and 94 (46%) respectively. No significant difference between the means number resistant and susceptible organisms (P = 0.1). Resistance patterns of E. coli isolates showed that ampicillin and amoxycillin have the highest single antibiotics resistance with 11 (12.5%) and 10 (11.4%) number of isolates respectively. Multiple antibiotic resistance pattern shows high frequency of occurrence in AMP, SXT, AML with 6 (6.82%), AMP, SXT with 5 (5.68%) and SXT, AML with 4 (4.55%) number of isolate with the pattern. Highest number of antibiotics resistance pattern of 7 antibiotics in occurred in 1 (1.14%) isolate with resistance phenotype include; OFX, AMP, SXT, CN, AML, CIP, TE. Two local plants Momordica charantia Linn and Aeschynomene uniflora E. May extracts and fractions were tested for activity on the E. coli isolates. The leaves and the whole plant yielded the highest extract weight in both plants. Thus Momordica charantia yield 40.60g and 38.35g for MOL and MOW, while Aeschynomene uniflora yield 37.90g and 24.19g for AEL and AEW respectively. Aqueous and ethanol extracts have the highest weight of 48.25g and 36.79g in Momordica charantia and 36.33g and 36.38g in Aeschynomene uniflora respectively. The petroleum ether in both plants showed the least weight extract obtained with weight of 09.84g and 08.92g respectively. Preliminary phytochemical screening of the plants extracts include test for carbohydrates (Molish‘s test and Fehling‘s test), alkaloid test (Mayer‘s test, Wagner‘s test and Dragendoff‘s test), tannins (Fe3Cl2 test), glycosides (Fehlings test), cardiac glycosides (Kella-killiani test), steroids (Salkwoski test), saponins (frothing test and emulsion test), flavonoid (Shinoda test), resin (copper acetate test) and
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anthraquinone (Borntrager‘s test). There was no significant difference between the mean weights of the albino mice before and after treatment (P=0.65 14 df) on pharmacological test. No death occurred of any mice after the 3 days experiment of haematological screening of albino mice blood after two weeks experiment on active MOL5 and AEW3 doses of methanol extract. The results of doses concentrations of 1000 mg/kg/day and 1600 mg/kg/day are not significantly different from the control, while 2900 mg/kg/day and 5000mg/kg/day results are significantly different from the control (P=0.05). Susceptibility pattern of E. coli isolates on extracts of Momordica charantia and Aeschynomene uniflora showed that ethanol and methanol proved to be effective for extraction of these plants having wider zones of inhibition of 18±2 mm and 18±1mm respectively. The isolates were more susceptible to leaves and whole plant extracts in both plants generally. Concentration has no significance effects in all the extracts tested (P=0.05). In Momordica charantia the isolates were more susceptible to leaves (methanol and ethanol) and whole plant (aqueous) with number susceptible of 52 (80%), 50(77%) and 48(74%) respectively, while more resistant to the root (methanol and petroleum ether extracts) with number of 53(82%) each. The organisms are more susceptible to whole plant and leave Aeschynomene uniflora methanol extracts with number susceptible of 49(75%) and 47(72%) respectively while resistant to the roots petroleum ether extract with number of 55(85%). All active extracts yielded the same MIC value of 62.5mg/ml except the methanolic extracts of MOL, MOS, MOW, AEL, AES and AEW with 31.2mg/ml. Low MBC of 31.2 mg/ml was obtained for MOL, AEL and AEW ethanolic extracts, MOL and AER for aqueous extracts, MOL, MOW, AEL and AEW for methanol extracts. Nine (9) TLC fractions were obtained from MOL methanol extract with Rf values ranging from 0.00 to 0.91 with solvent front of 7.9cm. Fraction number 5 (MOL5 deep purple) was the most active. MOL5 has Rf value of 0.51 and yielded 470mg (17.45%). Only three (3) fractions were obtained from the TLC of most active extract of Aeschynomene uniflora whole plant - AEW aqueous extract with solvent front of 7.8cm. Fraction number 3 (AEW3 green) showed more activity against the E. coli isolates with Rf value of 0.91 and yielded 370mg (46.84%). MOL5 fraction was more active than AEW3 fraction with zones of inhibition of 17±1 and 14±2 respectively. Zones of inhibitions did not differ significantly (P=0.05) from the values of E. coli (NTCT 10418) control. MOL5 showed MIC and MBC of 30μg/ml and 40μg/ml respectively, while AEW3 showed 40μg/ml in each case. Further phytochemicals screening revealed that each fraction contains 4 organic compounds. Steroids were found in MOL5 and more tannins than in AEW3. While AEW3 had more saponins compared to MOL5. This study revealed high prevalence of 34.81% infants‘ diarrhoea cases in Kano state. National policy on drugs disbursement, retail and usage should be observed strictly to avoid the antibiotics resistance shown by the Escherichia coli strains/isolates.
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CHARACTERIZATION AND SUSCEPTIBILITY OF ESCHERICHIA COLI ISOLATED FROM INFANTS DIARRHOEA TO MOMORDICA, CHARANTIA AND AESCHYNOMENE UNIFLORA EXTRACTS