Asian Pacific Journal of Tropical Medicine

594-601

Authors: Khaled A Abdelshafeek, Adnan A Elgattar, Aziza H Zarkoon, Mohamed A Alwahash, Abdelaaty A Shahat

Objective To determine the chemical composition of essential oils and lipid constituents of Ballota andreuzziana (B. andreuzziana), Teucrium zanonii (T. zanonii) and Verbena tenuisecta (V. tenuisecta) growing in Libya, and to test the antibacterial activity of different extracts of Teucrium zanonii. Methods The volatile oils of all plants were extracted by hydrodistillation method and analyzed by GC/MS method. The lipid constituents of plants were obtained by extraction with petroleum ether and fractionated into fatty alcohols, fatty acids and unsapoinfiable matters. Antibacterial activity of T. zanonii extracts and antioxidant activity of different extracts of T. zanonii were also studied. Results The volate oil of B. andreuzziana was found to consists of 20 compounds in which caryophyllin is the main one (63.1%), the volatile oil of T. zanonii consists of 74 compounds in which germacrene-D was the main compound, while the volatile oil of V. tenuisecta consists of 13 compounds with 1-octen-3-ol as a major constituent (52.87%). The study of antimicrobial activity of different extracts of V. tenuisecta showed that, both methanol and butanol extracts exhibited the highest activity against Mycobactirium phlei (M. phlei) and Candida albicans (C. albicans) respectively, while petroleum ether, fatty alcohols and unsaponifiable fractions had no antimicrobial activity against all the tested microorganisms. Investigation of the antioxidant activity of different extracts of T. zanonii using DPPH method proved that, the ethyl acetate and butanol fractions showed the highest activity where the inhibition percentage (I%) are 93.6 and 92.1 respectively. Conclusions This is the first report about the volatile oils of these plants where T. zanonii have the highest content and the highest number of the identified compounds. The study of antioxidant T. zanonii extracts proved that, the ethyl acetate, butanol and aqueous extracts have the highest antioxidant activity. Methanol and butanol extracts of V. tenuisecta exhibited the highest activity against M. phlei and C. albicans respectively.

605-609

Authors: Shyamapada Mandal

Objective To determine the larvicidal and adult emergence inhibition activities of castor (Ricinus communis) seed extract against three potential mosquito vectors Anopheles stephensi (An. stephensi), Culex quinquefasciatus (Cx. quinquefasciatus) and Aedes albopictus (Ae. albopictus) in India. Methods The R. communis seed extract was tested, employing WHO procedure, against fourth larval instars of the three mosquito species for 24 h and larval mortalities were recorded at various concentrations (2-64 μg/mL); the 24 h LC50 values of the R. communis seed extract were determined following Probit analysis. The larval killing, antipupation and adult emergence inhibition rates of the test extract, using a single concentration of 2 μ LC50, were studied at different time periods (24-72 h); the extract toxicity was tested against a fish, Oreochromis niloticus (O. niloticus). Results The R. communis seed extract exhibited larvicidal effects with 100 % killing activities at concentrations 32-64 μg/mL, and with LC50 values 7.10, 11.64 and 16.84 μg/mL for Cx. quinquefasciatus, An. stephensi and Ae. albopictus larvae, respectively. When the larvae were treated with the extract at a single concentration of 2×LC50, significant differences were observed, compared to control groups, in rate of pupation (P<0.001) as well as in adult formation (P<0.001). Conclusions The present findings suggest that the R. communis seed extract provided an excellent potential for controlling An. stephensi, Cx. quinquefasciatus and Ae. albopictus mosquito vectors.

614-618

Authors: Karthik Thanikachalam, Marimuthu Kasi, Xavier Rathinam

Objective To explore antidiabetic principles of the Eastern Nigeria mistletoe, Loranthus micranthus Linn. parasitic on Persea Americana. Methods The weakly acidic fraction of the aqueous methanol extract of the leaves of Loranthus micranthus (Linn.) was isolated and tested for its antidiabetic activities. The isolation of the weakly acidic fraction was carried out following established physico-chemical based procedures. Furthermore, alloxan-induced diabetic rats were treated intraperitoneally (ip) with 250 mg/kg and 400 mg/kg of the weakly acidic fraction, glibenclamide 10 mg/kg (positive control) and 2 mg/kg of 3 % v/v tween 20 (negative control). The sugar levels of the treated and untreated animals were determined by withdrawing the blood at regular intervals and testing them with an automated glucometer. The phytochemical analysis of the acidic fraction was carried out using standard procedures. Chromatographic techniques were employed in the subsequent isolation and purification of the constituents of the weakly acidic fraction. Results It was shown that the maximum effect of the weakly acidic fraction was obtained at 24 hours after administration and was found to be statistically comparable with that of the positive control. The phytochemical analysis revealed the presence of steroids, terpenoids, alkaloids, flavonoids, glycosides, carbohydrates, saponins, and acidic compounds in the crude extract and carbohydrates, flavonoids, terpenoids and oil in the weakly acidic fraction. Further purification of the weakly acidic fraction of the methanol extract using thin layer chromatography shows that toluene: methanol: diethyl amine (3:1:1) and chloroform: methanol: diethyl amine (9:1:1) are the best solvent system for the isolation of the various components of the weakly acidic fraction of the crude methanol extract of Loranthus micranthus. Conclusions The present study has led to the conclusion that the weakly acidic fraction of the plant under study has the potent antidiabetic activity and that the various components can be isolated using basic chromatographic techniques.

629-632

Authors: Selvan Nallaiyan, Ranjit Singh Amirtha Jacob Abbadorai, Sukumar Sundaramoorthy, Jeyakumar Nelson, Suryanarayana Veluvarthy Venkata Sanyasi

Objective To discuss the possibility of expressing the haemagglutinin-neuraminidase (HN) protein in prokaryotic system such as Escherichia coli (E. coli) cells by cloning the full length HN gene. Methods The full length HN gene of Newcastle Disease Virus (NDV) of size 1 734 bp was preciously isolated by RT-PCR. The sequence was assessed and submitted to Nucleic Acid Databank (NCBI) and the gene ID was EU215390.1 after cloning and sequencing. Now the assessed HN gene was subcloned into pET 32 a+ expression vector for production the HN protein in E. coli, BL21 (DE3) PLYSS cells following standard protocols. The crude lysate protein from the induced positive clone was size assessed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and their haemagglutination (HA) property against chicken RBC was assessed by standard micro HA test. Results The molecular size of the full HN gene of NDV as assessed by cloning and digesting the positive clone to release the insert was 1.7 kb. The expressed protein in both crude and pure form was assessed to be 63 kDa and 81 kDa, respectively. The HA activity of the crude protein of the positive clone was 1 in 40. Conclusions This finding indicates that the fusion protein retains the biological activity of native protein in the crude form and therefore could be used as a diagnostic reagent for antibody detection and for routine assessment of immune status in commercial layer forms.

637-641

Authors: Shyamapada Mandal, Manisha DebMandal, Nishith Kumar Pal

Objective To determine the antibiotic resistance of Vibrio cholerae (V. cholerae) O1 biotype El Tor serotype Ogawa isolates involved in an outbreak of watery diarrhea in Kolkata, and to explore the role of plasmid in mediating antibiotic resistance. Methods Antibiotic susceptibility and minimum inhibitory concentration (MIC) values of antibiotics for the isolated V. cholerae O1 Ogawa (n=12) were determined by disk diffusion and agar dilution methods, respectively, using ampicillin (Am), chloramphenicol (C), trimethoprim (Tm), tetracycline (T), erythromycine (Er), nalidixic acid (Nx), ciprofloxacin (Cp), amikacin (Ak) and cefotaxime (Cf). Plasmid curing of multidrug resistant (MDR) V. cholerae O1 Ogawa strains was done following ethidium bromide treatment. Following electrophoresis, the plasmid DNAs, extracted from the isolated MDR V. cholerae O1 Ogawa strains and their cured derivatives, were visualized and documented in ‘gel doc’ system. Results The outbreak causing V. cholerae O1 Ogawa isolates were MDR as determined by disk diffusion susceptibility test, and MIC determination. The isolates showed three different drug resistance patterns: AmTmTErNx (for 6 isolates), TmTErCp (for 5 isolates), and AmTmNx (for one isolate), and showed uniform sensitivity to C, Ak and Cf. The loss of plasmids with the concomitant loss of resistance to Am, Tm, T and Er of the isolates occurred following ethidium bromide treatment. Conclusions The current findings suggest that the V. cholerae O1 Ogawa associated with the cholera outbreak were MDR, and resistance to Am, Tm, T and Er among the isolates were plasmid mediated.

647-650

Authors: Suryanaryana Murty Upadhayula, Srinivasa Rao Mutheneni, Sriram Kumaraswamy, Madhusudhan Rao Kadiri

Objective To assess the prevalence of disease and microfilaraemia in villages of Karimnagar and Chittoor districts. Methods Data on age and sex of all the respondents were collected and compared with the microfilaria rate (mf) and density (20 μL of peripheral blood by using finger prick method) to examines the relationship between the dynamics of Wuchereria bancrofti among the population. Results Microfilaraemia prevalence was found among all the age groups and its occurrence was more prevalent especially above 30 years age groups. Similarly, the microfilaraemia and disease rates were found significantly higher in males compared to females. Conclusions Using these baseline data would be useful in planning for the elimination of lymphatic filariasis in Andhra Pradesh as per the WHO goal to eliminate lymphatic filariasis by 2020.

655-661

Authors: S Prakash, SL Sasikala, V Huxley J Aldous

Objective To isolate the multiple drug resistance (MDR) Mycobacterium tuberculosis (M. tuberculosis) and to screen for the bioactive compounds extracted from marine microalgae. Methods Sixty seven collected samples that were confirmed by 16S rDNA analysis as positive with M. tuberculosis infection were subjected to sensitivity test against commercially used front line and second line drugs by absolute concentration method using LJ slants. Seven bacterial isolates numbered I-78, I-101, I-127, I-173, I-202, I-262, I-327 showed resistant for more than 3 drugs were considered as MDR M. tuberculosis. Thus the percentage of 10.4 were recorded MDRTB. Fifteen marine micro algal extracts were screened for antimycobacterial activity and partial characterization of the active principles was done. Results The percentage contribution of marine micro algal species on the extraction of antimycobacterials indicated Isochrysis galbana (I. galbana) contain rich bioactive compounds and accounted for 60% inhibition of the total isolates. The percentage contribution of solvents on the extraction of antimicrobials from I. galbana showed that the methanol, chloroform, n-Butanol showed maximum of extraction. The purified eluted compounds (Rf 0.43) from TLC plate were chromatographed by gas chromatography. The eluted sterol compounds showed 13 unsaturated sterols with 3 major sterols. Conclusions The present study indicates the presence of unsaturated fatty acids may have the effect on MDR M. tuberculosis, indicating a potential natural alternative to antibiotics.

666-668

Authors: Boyi Ngwai Yakubu, KS Izebe, IG Ijele, D Ishaleku, US Inyang

Objective To detect IgG antibody to Chlamydophila pneumoniae (CP) in sera of HIV/AIDS patients and provide rationale for inclusion of routine screening for anti-CP antibodies and anti-chlamydial agents in the Nigerian National HIV/AIDS Management Plan. Methods Serum samples from 34 consenting HIV/AIDS patients attended a Government-approved Antiretroviral Treatment Facility in Abuja were screened by enzyme-linked immunosorbent assay for anti-CP IgG antibody using ImmunoComb® Chlamydia Bivalent IgG Test kit (Orgenics, Israel). Results Anti-CP IgG antibody was detected in 20 (58.8%) of 34 patients tested. The detection rate was higher among the males (8/13; 61.5%) than the females (12/21; 57.1%). Patients of the age group 16-30 years had the highest (7/10; 70%) detection of anti-CP IgG antibody. Conclusions The result of the present study suggests the presence of anti-CP antibodies in sera of the HIV/AIDS patients, and reinforces the need for routine screening for anti-CP antibodies as a necessary intervention to reduce the burden of Chlamydophila pneumoniae (C. pneumoniae) infections and to reduce HIV-positive morbidity in Nigeria. The outcome of this study also provides justification for the possible inclusion of anti-chlamydial agents in the National HIV/AIDS Management Plan to provide prophylaxis against or treat active C. pneumoniae infections.

671-672

Authors: MML Okome-Nkoumou, M Ondounda, A Dzeing-Ella, D Mounguengui, MI Nziengui Madjinou, C Magne, JR Nzenze

Neurocysticercosis (NCC) is recognized as a cause of neurologic disease worldwide. We reported two cases (one co-infected with the HIV) of NCC. Medical treatment led to recovery. NCC should be considered in tropical countries as a cause of epilepsy and included in the diagnosis of neurologic infections in HIV patients.