Asian Pacific Journal of Tropical Biomedicine

Issue 4,2014 Table of Contents

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  • 1  Purification, characterization and production optimization of a vibriocin produced by mangrove associated Vibrio parahaemolyticus
    Baskar Balakrishnan Jayappriyan Kothilmozhian Ranishree Sathish Thadikamala Prabakaran Panchatcharam
    2014(4):253-261. DOI: 10.12980/APJTB.4.2014C947
    [Abstract](19) [HTML](0) [PDF 907.40 K](85)
    Abstract:
    Objective: To identify a potential bacterium which produces antimicrobial peptide (vibriocin), and its purification, characterization and production optimization. The bacteria subjected in the study were isolated from a highly competitive ecological niche of mangrove ecosystem. Methods: The bacterium was characterized by phenotype besides 16S rRNA gene sequence analysis. The antibacterial activity was recognised by using agar well diffusion method. The vibriocin was purified using ammonium sulphate precipitation, butanol extraction, gel filtration chromatography, ion-exchange chromatography and subsequently, by HPLC. Molecular weight of the substance identified in SDS-PAGE. Production optimization performed according to Taguchi’s mathematical model using 6 different nutritional parameters as variables. Results: The objective bacterium was identified as Vibrio parahaemolyticus. The vibriocin showed 18 KDa of molecular mass with mono peptide in nature and highest activity against pathogenic Vibrio harveyi. The peptide act stable in a wide range of pH, temperature, UV radiation, solvents and chemicals utilized. An overall ~20% of vibriocin production was improved, and was noticed that NaCl and agitation speed played a vital role in secretion of vibriocin. Conclusion: The vibriocin identified here would be an effective alternative for chemically synthesized drugs for the management of Vibrio infections in mariculture industry.
    2  Quantitative estimation of hesperidin by HPTLC in different varieties of citrus peels
    Prawez Alam Aftab Alam Md. Khalid Anwer Saleh I Alqasoumi
    2014(4):262-266. DOI: 10.12980/APJTB.4.2014C1007
    [Abstract](24) [HTML](0) [PDF 611.12 K](90)
    Abstract:
    Objective: To develop a simple, selective, sensitive and accurate high-performance thin layer chromatography (HPTLC) method to determine the quantity of hesperidin in different varieties of citrus fruits. Methods: The method was carried out in aluminum-backed silica gel 60 F254 plates with ethyl acetate-methanol-water 15:3:2 (%, v/v) as mobile phase. Results: A compact band was obtained for hesperidin at Rf value of (0.40依0.04). The calibration plot was linear in the range of 100-800 ng/spot of hesperidin and the correlation coefficient of 0.9986 was indicative of good linear dependence of peak area on concentration. Limit of detection (8.87 ng/spot), limit of quantification (23.21 ng/spot), accuracy (less than 2%) and recovery (ranging from 98.55-99.38) were found satisfactory. Conclusions: The method developed can be used for routine analysis of hesperidin in crude drug as well as in herbal and pharmaceutical dosage form containing citrus fruits as an ingredient.
    3  New flavonoids from bioactive extract of Algerian medicinal plant Launeae arborescens
    Khaled Sekkoum Nasser Belboukhari Abdelkrim Cheriti
    2014(4):267-271. DOI: 10.12980/APJTB.4.2014C708
    [Abstract](29) [HTML](0) [PDF 321.88 K](81)
    Abstract:
    Objective: To investigate the butanol fraction of the water/acetone extract and isolate of the new flavonoids from Launeae arboescens. Methods: The compounds were isolated by liquid chromatographic methods and their structures were identified by using spectroscopic analysis. Results: The isolated compounds were identified as: 7-O-[α-rhamnopyranosyl 4',5,6-Trihydroxy flavone 1,4’,5’-Di-Methoxy 7-(5’’-Me Hexan)1-oyl flavanone 2, 3”-isopropyl pyrano [1”:7,4”:6] 3’,4’,5’,5-Tetrahydroxy flavanone 3,5,4’,5’-Tri-Hydroxy 7-(3’’-Me butan) -yl flavanone 4, 5,7-Dihydroxy-2',4',5' –trimethoxy-isoflavanone 5,5,6,7,4'-tetrahydroxy flavonol 6,7-O- [α-rhamnopyranosyl-(1->6)-β-glucopyranosyl]- 4',5,7-tri-hydroxy-flavanone 7,7-O-[α- rhamnopyranosyl-(1->6)-β-glucopyranosyl] 3',5-Dihydroxy 4’-Methoxy flavanone 8. Conclusions: The presence of different types of bioactive flavonoids in Launeae arboescens extract can explain the large ethnopharmacological uses and the potential activity of this medicinal plant.
    4  Effect of methanolic extract of Tetrapleura tetraptera (Schum and Thonn) Taub leaves on hyperglycemia and indices of diabetic complications in alloxan-induced diabetic rats
    Sunday Ene-Ojo Atawodi Ojochenemi Ejeh Yakubu Mubarak Labaran Liman Dorothy Uju Iliemene
    2014(4):272-278. DOI: 10.12980/APJTB.4.2014C73
    [Abstract](24) [HTML](0) [PDF 503.51 K](115)
    Abstract:
    Objective: To investigate the ameliorative role of Tetrapleura tetraptera (Schum and Thonn) Taub (T. tetraptera) leaf in hyperglycemia with associated conditions like oxidative stress, kidney damage and disorders in lipid metabolism. Methods: Five groups of five rats each intraperitoneally received the following treatment schedules for 7 d: untreated normal control, untreated alloxan-diabetic control, diabetic treated with glibenclamide, normal rats treated with extract (50 mg/kg) and diabetic rats treated with the extract. Evaluations were made for fasting blood sugar, body weight changes, malondialdehyde, aspartate aminotransferase, alanine aminotransferase, bilirubin, superoxide dismutase, catalase, lipid profile, packed cell volume, hemoglobin, urea and creatinine in all the rats. Results: Whereas the untreated diabetic rats showed a significant decrease (P<0.05) in packed cell volume, superoxide dismutase, catalase and high-density lipoprotein-cholesterol with a concomitant increase in the levels of malondialdehyde, fasting blood sugar, aspartate aminotransferase, alanine aminotransferase, bilirubin, urea and creatinine, administration of methanolic extract of T. tetraptera leaf or glibenclamide alleviated these altered parameters in the treated rats. Conclusions: Methanolic extract of T. tetraptera leaves possesses a potent capacity for treatment of diabetes and the accompanying complications, including oxidative stress and hyperlipidemia.
    5  Analgesic, antibacterial and central nervous system depressant activities of Albizia procera leaves
    Mst. Mahfuza Khatoon Mst. Hajera Khatun Md. Ekramul Islam Mst. Shahnaj Parvin
    2014(4):279-284. DOI: 10.12980/APJTB.4.2014C348
    [Abstract](33) [HTML](0) [PDF 282.28 K](85)
    Abstract:
    Objective: To ascertain analgesic, antibacterial and central nervous system (CNS) depressant activities of ethyl acetate, dichloromethane and carbon tetrachloride fractions of methanol extract of Albizia procera (A. procera) leaves. Methods: Leaves extracts of A. procera were tested for analgesic activity by acetic acid induced and formalin test method in mice. The in vitro antibacterial activity was performed by agar well diffusion method. CNS depressant activity was evaluated by hole cross and open field tests. Results: All the extracts at 200 mg/kg exhibited significant (P<0.01) analgesic activity in acetic acid induced and formalin tests method in mice. Analgesic activity of the ethyl acetate fraction was almost same like as standard drug indomethacin in acetic acid induced method. The CNS depressant activity of the extracts at 500 mg/kg was comparable to the positive control diazepam as determined by hole cross and open field test method. The extracts exhibited moderate antimicrobial activity against all the tested microorganisms (Staphylococcus aureus, Bacillus cereus, Pseudomonas aeruginosa, Esherichia coli, Shigella soneii, Shigella boydii) at concentration of 0.8 mg/disc. The measured diameter of zone of inhibition for the extracts was within the range of 7 to 12 mm which was less than the standard kanamycin (16-24 mm). Conclusions: It is concluded that all the extracts possess potential analgesic and CNS depressants activity. This study also showed that different fractions of methanol extract could be potential sources of new antimicrobial agents.
    6  Phytochemical screening and anticonvulsant studies of ethyl acetate fraction of Globimetula braunii on laboratory animals
    Musa Mumammad Aliyu Abdullahi Isma’il Musa Muhammad Ja’afar Kamal Magaji Garba Mohammed
    2014(4):285-289. DOI: 10.12980/APJTB.4.2014C925
    [Abstract](22) [HTML](0) [PDF 245.04 K](84)
    Abstract:
    Objective: To investigate the phytochemical properties and the anticonvulsant potential of the ethyl acetate soluble fraction of ethanol leaf extract of Globimetula braunii, a plant used in ethnomedicine for the treatment of epilepsy. Methods: The phytochemical screening was carried out using standard protocol while the anticonvulsant activity was studied using maximal electroshock test in chicks, pentylenetetrazole and 4-aminopyridine-induced seizures in mice. Results: The preliminary phytochemical screening carried out on the crude ethanol extract revealed the presence of saponins, carbohydrates, flavonoids, tannins, anthraquinones and steroids. Similarly, tannins, flavonoids and steroids/terpenes were found to be present in the ethyl acetate fraction. In the pharmacological screening, 150 mg/kg of the fraction protected 83.33% of animals against pentylenetetrazole-induced seizure in mice whereas sodium valproate a standard anti-epileptic drug offered 100% protection. In the 4-aminopyridine-induced seizure model, the fraction produced a significant (P<0.05) increase in the mean onset of seizure in unprotected animals. The fraction did not exhibit a significant activity against maximal electroshock convulsion. The median lethal dose of the fraction was found to be 1 261.91 mg/kg. Conclusions: These results suggest that the ethyl acetate fraction of Globimetula braunii leaves extract possesses psychoactive compound that may be useful in the management of petit mal epilepsy and lend credence to the ethnomedical use of the plant in the management of epilepsy.
    7  In vitro antioxidant and cytotoxic activities of essential oil of Feronia elephantum Correa
    Ramaraj Thirugnanasampandan Delma David
    2014(4):290-293. DOI: 10.12980/APJTB.4.2014B878
    [Abstract](27) [HTML](0) [PDF 294.33 K](89)
    Abstract:
    Objective: To analyse the chemical composition and evaluation of antioxidant, cytotoxic and DNA fragmentation activities of essential oil of Feronia elephantum Correa. Methods: Chemical composition analysis of hydrodistilled essential oil was determined by gas chromatography-mass spectrometry and in vitro antioxidant activity of oil was determined by DPPH free radical, hydroxyly radical scavenging, metal chelating and prevention of deoxyribose degradation. Cytotoxicity and DNA fragmentation activities against breast cancer cells (MCF-7) were also analyzed. Results: Gas chromatography-mass spectrometry analysis revealed the presence of 24 compounds with caryophyllene oxide (62.29%) as major compound. A considerable antioxidant, cyotoxic and DNA fragmentation activities of oils was observed. Conclusions: The result of this study clearly indicates oil could be useful for food preservation and preparation.
    8  In vitro anti oxidant activity and acute oral toxicity of Terminalia paniculata bark ethanolic extract on Sprague Dawley rats
    Ramgopal Mopuri Balaji Meriga
    2014(4):294-298. DOI: 10.12980/APJTB.4.2014C1068
    [Abstract](20) [HTML](0) [PDF 263.05 K](83)
    Abstract:
    Objective: To ensure the safety and evaluate the anti oxidant activity of Terminalia paniculata (T. paniculata) ethanolic extract in Sprague Dawley rats. Methods: The solvent extracts (hexane, ethyl acetate and ethanol) of T. paniculata were subjected to phytochemical analysis and their DPPH radical scavenging activity was assayed. The oral acute toxicity was evaluated using ethanolic extract of T. paniculata. Results: Ethyl acetate and ethanolic extracts showed more phytochemicals, whereas highest DPPH scavenging activity was found in ethanolic extract. In an acute toxicity study, T. paniculata ethanolic extract was orally administered (1 000 mg/kg body weight) to rats and observed for 72 h for any toxic symptoms and the dose was continued up to 14 d. On the 15th day rats were sacrificed and blood samples were collected from control and test animals and analyzed for some biochemical parameters. We did not observe any behavioral changes in test groups in comparison with their controls. Also, there were no significant alterations in biochemical, hematological (hemoglobin content and blood cells count) and liver function parameters such as serum glutamate pyruvate transaminase, serum glutamate oxaloacetate transaminase, alkaline phosphatase, total proteins, albumin and bilirubin levels between T. paniculata ethanolic extract treated and normal control groups. Conclusions: Together our results demonstrated that T. paniculata ethanolic possessed potent antioxidant activity and it was safer and non toxic to rats even at higher doses and therefore could be well considered for further investigation for its medicinal and therapeutic efficacy.
    9  Phytochemical and in vitro biological investigations of methanolic extracts of Enhydra fluctuans Lour.
    Sourov Kuri Md. Mustahsan Billah S. M. Masud Rana Zannatul Naim Md. Mahmodul Islam Md. Hasanuzzaman Md. Ramjan Ali Rana Banik
    2014(4):299-305. DOI: 10.12980/APJTB.4.2014C677
    [Abstract](17) [HTML](0) [PDF 506.35 K](83)
    Abstract:
    Objective: To study the phytochemical and biological properties (antioxidant, anthelmintic and thrombolytic) of methanolic extracts of Enhydra fluctuans Lour., a plant belonging to the Asteraceae family. Methods: The phytochemical evaluation was carried out by qualitative analysis. In vitro antioxidant activity of extract was studied using free radical scavenging assay, ability of reduction, total phenol and total flavonoid contents determination assays. The anthelmintic activity was determined using paralysis and death time of Pheretima posthuma (earthworm) and thrombolytic activity by clot disruption assay. Results: The phytochemical evaluation showed significant presence of flavonoids, triterpenes, carbohydrate, reducing sugars, saponins, phenols, diterpenes, protein and tannin. The antioxidant activity was found significant [IC50=(135.20±0.56) µg/mL] as compared to ascorbic acid [(130.00±0.76) µg/mL]. The reducing power was increased with concentration. Total phenol and total flavonoid contents were (153.08±0.38) mg/mL and (172.04±0.56) mg/mL respectively. The paralysis and death time of earthworms for different concentrations of extract were determined and compared with albendazole. The results showed that 10 mg/mL of the crude extract had similar effect with albendazole. Additionally, the crude extract showed a concentration depended relationship with its anthelmintic property. The clot lysis activity of crude extract was compared to the standard streptokinase’s clot lysis (40.13%) activity and found significant (31%). Conclusions: The study proves that the crude methanolic extract of Enhydra fluctuans Lour. has significant antioxidant, anthelmintic and thrombolytic activity containing wide range of phytochemicals.
    10  A cross sectional study on antibiotic resistance pattern of Salmonella typhi clinical isolates from Bangladesh
    Adnan Mannan Mohammad Shohel Sultana Rajia Niaz Uddin Mahmud Sanjana Kabir Imtiaj Hasan
    2014(4):306-311. DOI: 10.12980/APJTB.4.2014C770
    [Abstract](21) [HTML](0) [PDF 388.98 K](88)
    Abstract:
    Objective: To investigate and compare the resistance and sensitivity of Salmonella typhi samples to commonly used antibiotics in three major divisions of Bangladesh and to evaluate the gradually developing resistance pattern. Methods: The antibiotic susceptibility of 70 clinical isolates collected from blood, sputum, urine and pus samples were identified by specific antisera and with standard biochemical tests. The patients were divided into 5 age groups. Susceptibility and resistance was also tested by Kirby-Bauer disc diffusion method using 12 regularly used antibiotics. Results: Antibiotic susceptibility test demonstrated that 64.28% isolates of Salmonella typhi were multidrug resistant. Present study suggests that the clinical samples were mostly resistant against nalidixic acid with all age groups and in all three divisions with similar resistance pattern. Resistance is more common among adult people (30-40 years) and children (0-10 years). Salmonella typhi was mostly sensitive against gentamycin, chloramphenicol and ciprofloxacin. Conclusions: Although the population density of Dhaka region is markedly higher than Rajshahi and Chittagong regions, no significant difference in resistance pattern was found. The rate of multidrug resistance is a matter of concern. Physicians should reconsider before prescribing nalidixic acid and cefixime. Further molecular study is needed to reveal the genomic and proteomic basis of resistance.
    11  Comparative susceptibility to permethrin of two Anopheles gambiae s.l. populations from Southern Benin, regarding mosquito sex, physiological status, and mosquito age
    Nazaire A?zoun Rock A?kpon Roseric Azondekon Alex Asidi Martin Akogbéto
    2014(4):312-317. DOI: 10.12980/APJTB.4.2014C1093
    [Abstract](23) [HTML](0) [PDF 589.95 K](82)
    Abstract:
    Objective: To investigate what kind of mosquito sample is necessary for the determination of insecticide susceptibility in malaria vectors. Methods: Larvae and pupae of Anopheles gambiae s.l. (An. gambiae) mosquitoes were collected from the breeding sites in Littoral and Oueme departments. The Centers for Disease Control and Prevention (CDC) susceptibility tests were conducted on unfed male and female mosquitoes aged 2-5 days old. CDC susceptibility tests were also conducted on unfed, blood fed and gravid female mosquitoes aged 2-5 days old. These susceptibility tests were also conducted on unfed and blood fed female mosquitoes aged 2-5 days old and 20 days old. CDC biochemical assay using synergist was also carried out to detect any increase in the activity of enzyme typically involved in insecticide metabolism. Results: Female An. gambiae Ladji and Sekandji populations were more susceptible than the males when they were unfed and aged 2-5 days old. The mortality rates of blood fed female An. gambiae Ladji and Sekandji populations aged 2-5 days old were lower than those obtained when females were unfed. In addition, the mortality rates of gravid female An. gambiae Ladji and Sekandji populations aged 2-5 days old were lower than those obtained when they were unfed. The mortality rate obtained when female An. gambiae Sekandji populations were unfed and aged 20 days old was higher than the one obtained when these populations were unfed and aged 2-5 days old. The results obtained after effects of synergist penicillin in beeswax on F1 progeny of An. gambiae Ladji populations resistant to permethrin showed that mono-oxygenases were involved in permethrin resistant F1 progeny from Ladji. Conclusions: The resistance is a hereditary and dynamic phenomenon which can be due to metabolic mechanisms like overproduction of detoxifying enzymes activity. Many factors influence vector susceptibility to insecticide. Among these factors, there are mosquito sex, mosquito age, its physiological status. Therefore, it is useful to respect the World Health Organization criteria in the assessment of insecticide susceptibility tests in malaria vectors. Otherwise, susceptibility testing is conducted using unfed female mosquitoes aged 3-5 days old. Tests should also be carried out at (25±2) ℃ and (80±10)% relative humidity.
    12  Effects of melatonin on changes in cognitive performances and brain malondialdehyde concentration induced by sub-chronic co-administration of chlorpyrifos and cypermethrin in male Wister rats
    Idris Sherifat Banke Ambali Suleiman Folorunsho Bisalla Mohammed Suleiman Mohammed Musa Onukak Charles Ayo Joseph Olusegun
    2014(4):318-323. DOI: 10.12980/APJTB.4.2014C55
    [Abstract](12) [HTML](0) [PDF 298.23 K](78)
    Abstract:
    Objective: To evaluate the ameliorative effect of melatonin on sub-chronic chlorpyrifos (CPF) and cypermethrin (CYP)-evoked cognitive changes in male Wistar rats. Methods: Fifty adult male Wistar rats, divided into five groups of ten rats each, were used for the study. Groups 1 and Ⅱwere given distilled water and soya oil (2 mL/kg) respectively. Group Ⅲ was administered with melatonin at 0.5 mg/kg only. Group Ⅳ was administered with CPF [7.96 mg/kg (1/10th LDResults: CPF and CYP induced cognitive deficits and increased brain malonaldehyde concentration, which were all ameliorated by melatonin. Conclusion: Cognitive deficits elicited by CPF and CYP was mitigated by melatonin due to its antioxidant property.
    13  The effects of exposure to pesticides on the fecundity status of farm workers resident in a rural region of Fars province, southern Iran
    Masoud Neghab Mohammadjafar Momenbella-Fard Reza Naziaghdam Narges Salahshour Maryam Kazemi Hamzeh Alipour
    2014(4):324-328. DOI: 10.12980/APJTB.4.2014C586
    [Abstract](15) [HTML](0) [PDF 491.94 K](84)
    Abstract:
    Objective: To investigate the prevalence of fecundity and other reproductive problems among a group of farmers in Kavar district of Fars province, southern Iran. Methods: A total of 268 randomly selected married male farm workers were investigated. A questionnaire was devised and validated [Cronbach’s α-coefficient (0.81)]. Subjects were directly interviewed and the questionnaire forms were completed for them. Results: The prevalence of current primary infertility among the studied population was about 7.4% (P=0.001). Similarly, 6.3% of farm workers had offsprings with congenital malformations. Finally, 1.5% and 9% of farmers’ wives had a history of stillbirth and abortion, respectively. It was concluded that the prevalence of current primary infertility were higher among farm workers families than in the normal population (P<0.05). Additionally, stillbirth and spontaneous abortion were more common in the wives of farm workers than in the normal population, although the difference did not reach statistical significance. Conclusions: These effects are likely to be attributed to the exposure of farm workers to pesticides.
    14  Theoretical and experimental study on lipophilicity and wound healing activity of ginger compounds
    Mohammed Afroz Bakht Mohammed F. Alajmi Perwez Alam Aftab Alam Prawez Alam Tariq Mohammed Aljarba
    2014(4):329-333. DOI: 10.12980/APJTB.4.2014C1012
    [Abstract](31) [HTML](0) [PDF 401.23 K](90)
    Abstract:
    Objective: To correlate the chromatographic and computational method to calculate lipophilicity of selected ginger compounds and to observe the effects of log P on wound healing. Methods: Mixtures of acetonitrile and water with acetonitrile content between 95% and 50% v/ v in 5% increments were kept separately in 10 different chromatographic chambers, saturated with solvent for 2 h. Spots were observed under UV light at λ=254 nm p-anisaldehyde used as a spraying reagent. Theoretical calculation was done using the Alogps 2.1 online program at www. vcclab.org/lab/alogps. For percentage wound contraction, five groups of animal (mice) (25-30 g) of either sex were selected. Wound were created on dorsal surface of animals using toothed forceps, scalpel and pointed scissors. The wound areas were calculated using vernier caliper. After making wound mice were orally administered 35 mg/kg 6-shogoal, 6-gingerol, 8-gingerol and 10- gingerol respectively. Group E as the control group received tap water. Results: The lipophilicity values determined in thin layer chromatography were correlated with the theoretically calculated various log P by linear regression analysis. Significant correlations were found between log P values calculated by software program and the experimental reversed-phase thin-layer chromatography data. Order of wound healing property of ginger compounds is directly dependent on lipophilicity i.e. more lipophilic compound has highest activity. Conclusions: Experimentally determined lipophilicity (RMO) values were correlated with log P determined by software's and found satisfactory. Lipophilicity (RMO) is a useful parameter for the determination and prediction of biological activity of ginger compounds.
    15  Extreme human annoyance caused by Ctenocephalides felis felis (cat flea)
    Mohammad Reza Youssefi Mohammad Taghi Rahimi
    2014(4):334-336. DOI: 10.12980/APJTB.4.2014C795
    [Abstract](29) [HTML](0) [PDF 565.35 K](86)
    Abstract:
    The authors report a case of human dermal complications by Ctenocephalides felis felis (cat flea) in a 28 years old woman who dwell in rural area in Mazandaran province, Iran. Furthermore, in the current report clinical manifestations, etiology and different aspects of infestation were discussed elaborately. And the importance of controlling fleas was highlighted as well.

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