Asian Pacific Journal of Tropical Medicine

930-933

Authors: Gang Lu, Xuchu Hu, Can Huang, Yajun Lu, Lixian Wu, Lihua Li, Jing Xu, Xinbing Yu

Objective To explore antischitosome effects of artemether, hemin and Fe3+ on Sj LDH. Methods Enzyme activity of rSj LDH was assayed in the standard reaction system by adding different concentration of reagents (0.00-0.10 mM artemether, 0.00-0.02 mM hemin, 0.00-0.50 mM Fe3+). Same solvents of the each reagent were used as control. Results There was no enzyme activity inhibition observed at 0.10 mM artemther; obivious inhibition for lactate oxidation reaction and pyruvate reduction reaction were detected at 0.002 mM and 0.004 mM of hemin, respectively; comparing with that of the control (P<0.05). The relative enzymatic activity inhibitions for pyruvate reduction reaction and lactate oxidation reaction at 0.02 mM hemin were 93.48% and 100.00%, respectively, comparing with that of the control (P<0.01); both pyruvate reduction and lactate oxidation reaction were inhibited completely at 0.50 mM Fe3+, comparing with that of the control (P<0.01). Conclusions The results implied that Sj LDH was not the direct molecular target of artemether. Hemin and Fe3+ are inhibitors of Sj LDH.

939-942

Authors: Yutaka Midorikawa, Satoshi Nakamura, Rattanaphone Phetsouvanh, Manivanh Vongsouvaht, Kaoru Midorikawa

Objective To know the current state of non-typhoid Salmonella infection in Laos. To examine the usefulness of new screening methods for Salmonella using citrus. Methods Non-typhoid Salmonella infection of person in Lao PDR was studied in this research (2004-2009). The site was Vientiane capital city in 2004. Research from rural villages locating suburb of Vientiane during 2005-2008 was carried out. Rural villages in Attapu province where ethnic minorities were living was searched for this study in 2009. During this research, to detect Salmonella strain, a new method using citrus and citrus extract named MY phenomenon that observing black ring (MIDO ring) on DHL agar was tried. The slice lemon and lime were used for this trial in 2004. After 2005, disk of ascorbic acid and citric acid were used for the device instead of citrus fruits itself. Results During this research, 65 of 272 human samples (23.9%) were infected with non-typhoid Salmonella. Conclusions During this study, the method using citrus and citrus extracts was accepted for the detection of Salmonella. This study shows that with citrus and citrus extract, detection of Salmonella is possible using only DHL media. Results suggest that infectious rate of non-typhoid Salmonella was high.

948-953

Authors: Chinnaperumal Kamaraj, Abdul Abdul Rahuman

Objective To determine the larvicidal and adulticidal activities of hexane, ethyl acetate and methanol extracts of Momordica charantia (M.charantia), Moringa oleifera(M. oleifera), Ocimum gratissimum (O. gratissimum), Ocimum tenuiflorum (O. tenuiflorum), Punica granatum(P. granatum) and Tribulus terrestris (T. terrestris) against Culex gelidus (Cx. gelidus) and Culex quinquefasciatus (Cx. quinquefasciatus). Methods Bioassay test was carried out by WHO method for determination of larvicidal and adulticidal activity against mosquitoes. Results All plant extracts showed moderate larvicidal and adulticidal activities, however the effective larval mortality was found in the leaf ethyl acetate and methanol extract of O. gratissimum and bark methanol extract of M. oleifera against Cx.gelidus with LC50 values of 39.31, 66.28, and 21.83 μg/mL respectively, and methanol extract of O. gratissimum, O. tenuiflorum and P. granatum against Cx. quinquefasciatus with LC50 values of 38.47, 24.90 and 67.20 μg/mL, respectively. The adult exposed for 1 h and mortality was recorded at 24 h recovery period. Above 90% mortality was found in the ethyl acetate and methanol extract of all experimental plants at the concentrations of 500 μg/mL. Conclusions The present results suggest that the medicinal plant extracts provided an excellent potential for controlling Cx. gelidus and Cx. quinquefasciatus.

957-960

Authors: Varaprasadham Irudayaraj, Janaky M, Marimuthu Johnson, Nallayan Selvan

Objective To screen the anti-cancer spike-mosses for the presence of various bioactivities and to identify the important bioactive chemicals present in Selaginella inaequalifolia (S. inaequalifolia) (Hook. & Grev.) Spring. Methods Preliminary phytochemical screening was done by following the method of Brindha et al. Antimicrobial study was carried out by disc diffusion method. Results Results of preliminary phytochemical screening on five different extracts (petroleum ether, benzene, chloroform, ethanol and distilled water) of the spike-moss S. inaequalifolia show the presence steroids, triterpenes, phenolic group, tannin, sugars and catechin. Alkaloids, amino acids, anthraquinone and reducing sugar did not show any positive result. Among the five different extracts, ethanol and chloroform extracts show the presence of maximum number (4 each) of compounds. The results on antimicrobial studies show that all the three microbes [Staphylococcus aureus (S. aureus), Escherichia coli (E. coli) and Candida albicans (C. albicans)] tested are resistant to the ethanol extract and susceptible to petroleum ether extract. The petroleum ether extract shows maximum inhibition with 45 mm of inhibition zone in C. albicans. The inhibition zone in S. aureus and E. coli are 26 mm and 22 mm respectively. Conclusions The present study shows S. inaequalifolia having potent antibacterial and anticandidal activities.

965-970

Authors: Gollapalle Lakshminarayana shastry Viswanatha, Santosh Kumar Vaidya, Ramesh C, Nandakumar Krishnadas, Srinath Rangappa

Objective The alcoholic extract of stem bark of Terminalia arjuna (ALTA) was screened for antioxidant and antimutagenic (anticlastogenic) activity. Methods Antioxidant property was determined by 1, 1, Diphyny l, 2-Picryl hydrazyl (DPPH) assay, super oxide radical scavenging activity, lipid peroxidation assay and total polyphenolic content was determined by Folin-Ciocalteau's reagent. Antimutagenic activity was evaluated using micronucleus test in mice. Results The ALTA has shown potent antioxidant activity with EC50 of 2.491±0.160, 50.110±0.150 & 71.000±0.250 in DPPH assay, superoxide radical scavenging activity and lipid peroxidation assay, which is comparable with ascorbic acid with EC50 of 2.471±0.140, 40.500±0.390 and 63.000±0.360 respectively. In micronucleus test, ALTA (100 & 200mg/kg, p.o.) showed significant reduction in percentage of micronucleus in both polychromatic erythrocytes (PCE) and normochromatic erythrocytes (NCE) and also shown significant reduction in P/N ratio. Conclusions These results suggested that ALTA possess significant antioxidant and antimutagenic activity.

976

Authors: Viroj Wiwanitkit

982-987

Authors: Jayanta M Sankhari, Ravirajsinh N Jadeja, Menaka C Thounaojam, Ranjitsinh V Devkar, Ramachandran AV

Objective To evaluate the safety of ethanolic seed extract of Eugenia jambolana (EJSE) using acute and sub-chronic toxicity assays in Swiss albino mice as per Organisation for Economic Co-operation and Development (OECD) guidelines. Methods Possible behavioral changes and lethality were observed in mice administered a single dose [1 000, 2 000, 3 000, 4 000 or 5 000 mg/kg body weight (BW)] of EJSE. Plasma levels of metabolic, hepatic, cardiac and renal function markers, electrolytes, blood count and histopathology of major organs were monitored in mice chronically treated with EJSE (1 000, 2 000 or 3    000 mg/kg BW) for 28 days. Results Since no mortality was recorded in the acute toxicity evaluation up to a dose of 5 000 mg/kg bodyweight of EJSE, 50% lethal dose (LD50) was assumed to be > 5 000 mg/kg BW. In the sub-chronic toxicity evaluation, no adverse observations were recorded in mice administered with 2 000 mg/kg BW of EJSE; however at 3 000 mg/kg BW dose, moderately significant increase in the plasma levels of urea and creatinine was observed. Hence, the lowest observable adverse effect level (LOAEL) for EJSE was found to be 3 000 mg/kg BW and the no observable adverse effect level (NOAEL) was adjudged as 2 000 mg/kg BW. Conclusions It can be concluded from this study that, orally administered EJSE is safe up to a 10 fold higher dose than its reported therapeutic dose.

993-996

Authors: Amuta EU, Houmsou RS, Mker SD

Objective To assess women's perceptions and risk factors that could expose them to intestinal parasitic infections in Makurdi, Benue State, Nigeria. Methods A total of 750 faecal samples were collected from women at different reproductive stages (pre-menstrual, menstrual and post-menstrual), and the faeces were tested by the formol ether concentration technique. Results A total of 426 (56.8%) samples were found positive for various intestinal parasites with hookworm (4.8%), Ascaris lumbricoides (9.3%), Taenia sp (2.1%), Entamoeba histolytica (18.9%) and Entamoeba coli (21.6%). Women at pre-menstrual and post-menstrual stages recorded higher prevalence rates with 72.8% and 63.9%, respectively. No significant difference in prevalence was observed between women at different reproductive stages and women infected by different parasites (χ2 = 30.6, P > 0.05). Sweet things, rotten fruits and improperly cooked meat were perceived as the causes of intestinal parasitic infections among the pre and post menstrual women. Sources of drinking water like river, well, water bought from vendors and patronizing food vendors were observed as risk factors contributing to the prevalence of intestinal parasitic infections among women. Factors like not washing hands before eating and after defaecation, use of leaves and ordinary papers for cleaning after defaecation were also observed to be contributing to the prevalence of intestinal parasitic infections. Conclusions Persuasive health education and rigorous hygiene measures should be employed in schools, maternity clinics and among the populace to reduce transmission and infection with intestinal parasites.

1001-1003

Authors: Pradeep K Singh

Small pox has been eradicated completely but its unique pathology and sequlae still come across in routine clinical practice. Osteoarticular changes following small pox infection has been elaborated in literature. We present a similar but rare patient with osteomyelitis variolosa and joint deformity involving elbow, wrist joints and humerus fracture. The condyles were typically elongated as central portion of distal humerus absorbed. Fracture united uneventfully following stabilization with dynamic compression plate and bone grafting. Patient showed satisfactory elbow function at the end of last follow-up.

1006-1008

Authors: Azikiwe CCA, Ifezulike CC, Amazu LU, Enye JC, Nwosu PJC, Osuala FN

We report two peculiar cases that were observed in two communities of Anambra State, South East Nigeria. The first case was a woman of 44 with a history of blood transfusion from a donor of questionable HIV status. A pint of blood was transfused in a private Hospital on the 15 th January, 2000. She has had no form of antiviral therapy until 22nd April, 2008 when she came to the hospital for routine medical check up. Her HIV result came out confirmed positive while that of her husband came out negative. She has had active sexual life with her husband in the previous eight years as they have had some difficulties in getting their desired extra two children having earlier gotten a boy and a girl. The second case involves a family of six. A couple with four children of ages about 6 months, 2 years, 3 years and 5 years. The youngest, a female presented with fever and multiple lymphadenopathy. She tested HIV positive and so was her mother and her second sibling. But her father and two other siblings remained negative at post three and six months intervals from their very first test date with us. These reported cases appear to go contrary to earlier formed opinion on HIV transmission. We support the opinion on a naturally, occurring substance, APOBEC3G which possibly confers permanent immunity against the HIV virus. We recommend clinical trials of the characterized APOBEC3G as a vaccine for non HIV infected persons of all ages and to people living with as a therapeutic drug.

846-850

Authors: RD Sharma, S Petare, GB Shinde, Goswami Kalyan, MVR Reddy

Objective To explore the effect of herbal polyphenolics on filariasis in vitro. Methods Two herbal extracts, methanolic extracts of roots of Vitex negundo Linn. (Nirgundi) and leaves of Aegle marmelos Juss. (Beal) in different concentrations ranging from 40-80 ng/mL were tested for their antifilarial activity either alone or in combination with diethyl carbonate (DEC) (300 μ g/mL) and/or H2O2 (0.5 mM). Results Combination of DEC and each extract had significant anti-filarial effect. And fractions of both extracts were not effective as crude herbal extract. Conclusion Such unique pharmacodynamics reported in this study might provide new drug development stratagem against filariasis.

855-859

Authors: Noppadon Tangpukdee, Khin Myat Wai, Sant Muangnoicharoen, Shigeyuki Kano, Nanthaporn Phophak, Janram Tiemprasert, Srivicha Krudsood, Polrat Wilairatana

Objective To illustrate the clinical features and investigate the indicators associated with a fatal outcome in adult patients with severe Plasmodium falciparum malaria admitted to the Hospital for Tropical Diseases, Bangkok, Thailand. Methods We studied 202 adult malaria patients admitted to the Intensive Care Unit. A total of 43 clinical variables were identified by univariate and logistic regression analyses, to eliminate confounding factors. Results Regarding the statistical methods, only 6 variables-jaundice, cerebral malaria, metabolic acidosis, body mass index, initial respiratory rate, and white blood cell count-were significant indicators of death, with adjusted odds ratios (95% CI) of 15.2 (2.1-32.3), 4.3 (2.3-12.6), 3.3 (2.3-5.7), 2.4 (1.9-3.5), 2.2 (1.5-2.6), and 1.7 (1.2-3.1), respectively. Conclusion Our study found that jaundice, cerebral malaria, metabolic acidosis, body mass index, initial respiratory rate and white blood cell count were indicators of fatal outcome in severe Plasmodium falciparum malaria. Further studies on the fatal indicators in severe malaria need to be compared with data from different geographical areas, to construct practical measures to address potentially fatal indicators in different settings.

866-870

Authors: N Mirzaie Damabi, AA Moazedi, SM Seyyednejad

Objective To investigate Petroselinum crispum Latifolum (parsley) for treating stomach and intestinal disorders in Iran. Methods An 80% ethanol extract was prepared from parsley seeds, and its antispasmodic activity assessed by measuring contractions of isolated ilea induced by 60 mM potassium chloride (KCl). A piece of ileum from an adult male Wistar rat was dissected and mounted in an organ bath containing Tyrode's solution, and ileum contractions were recorded by an isotonic transducer under one gram resting tension. The effect of parsley extract was measured in the presence of two antagonists' of α — and β —adrenoceptors. Results These experiments showed that parsley extract inhibited the response to 60 mM KCl in a concentration-dependent manner (P<0.01, n=7). The spasmolytic effect of parsley extract was unaffected by 1 μ M phentolamine or 1 μ M propranolol. Conclusion This study shows that parsley seed extract is a relaxant of isolated rat ileum and the relaxation effect of extract does not involved adrenergic receptors.

874-877

Authors: Marimuthu Govindarajan

Objective To determine the mosquito larvicidal activity of leaf essential oil and their chemical constituents from Clausena anisata(C. anisata) (Willd.) Hook. f. ex Benth. against Culex quinquefasciatus, Aedes aegypti and Anopheles Stephensi. Methods Essential oil was obtained by hydro-distillation and the chemical composition of the leaf essential oil was analyzed using gas chromatography-mass spectrometry. The mosquitoes were reared in the vector control laboratory and twenty late III instar larvae of three mosquito species were exposed to based on the wide range and narrow range tests, essential oil was tested at 50, 100, 150, 200 and 250 ppm and each compound was tested at various concentration (5-75 ppm) and were assayed in the laboratory by using the protocol of WHO 2005; the 24 h LC50 values of the C. anisata leaf essential oil and their major compounds were determined following Probit analysis. Results The oil contained were mainly β-pinene (32.8%), sabinene (28.3%), germacrene-D (12.7%), estragole (6.4%) and linalool (5.9%). The essential oil from the leaves of C. anisata exhibited significant larvicidal activity, with 24 h LC50 values of 140.96, 130.19 and 119.59 ppm, respectively. The five pure constituents extracted from the C. anisata leaf essential oil were also tested individually against three mosquito larvae. The LC50 values of β-pinene, sabinene, germacrene-D, estragole and linalool appeared to be most effective against Anopheles stephensi (LC50–23.17, 19.67, 16.95, 11.01, 35.17 ppm) followed by Aedes aegypti (LC50–27.69, 21.20, 18.76, 12.70, 38.64 ppm) and Culex quinquefasciatus (LC50–32.23, 25.01, 21.28, 14.01, 42.28). Conclusion The essential oil of C. anisata contains five major compounds and has remarkable larvicidal properties, which may be considered as a potent source for the production of natural larvicides.

884-886

Authors: LU Amazu, CCA Azikiwe, CJ Njoku, FN Osuala, PJC Nwosu, AO Ajugwo, JC Enye

Objective To scientifically verify the claims of our traditional healers on the anti-inflammatory activity of Carica papaya (C. papaya) and possibly deduce its activities. Methods 0.1 mL of fresh egg albumin was injected into the right hind-paw of adult white Wistar rats to induce inflammation an hour post intraperitoneal (IP) administration of 50-200 mg/kg doses of the extract to 3 groups of 5 rats per group. The 4th group of 5 rats was used as negative control and received 2 mL/kg (IP) of physiological saline, while the 5th group of 5 rats was used as positive-comparative control and received (IP) 150 mg/kg of aspirin. Increases in diameter of the paw were measured with the aid of Veneer Calipers before extract administration and at interval of 30 minutes post administration for further 2 hours. Percentage inhibition of oedema was calculated for each dose group and results were subjected to statistical analysis using student t-test and analysis of variance (ANOVA). Results All doses of extract showed a dose and time dependent inhibition effects of oedema (P<0.05). Conclusion This work is at present though limited to animals, the anti-inflammatory activity of the seeds of C. papaya is perhaps proven.

890-893

Authors: Rengasamy Ragupathi Raja Kannan, Rajasekaran Arumugam, Perumal Anantharaman

Objective To evaluate the antibacterial activity of Halophila stipulacea (H. stipulacea), Cymodocea serrulata (C. serrulata) and Halodule pinifolia (H. pinifolia) against seven human bacterial pathogens. Methods The antibacterial activities of the extracts on the various test organisms using disc diffusion method and Minimum Inhibitory Concentraction (MIC). Results Methanol and chloroform extracts of all the three seagrasses were active against all the tested pathogens, whereas the hexane extract of seagrasses was not active against Staphylococcus aureus (S. aureus). Antibacterial activity of three seagrass screened, was in the order of H. pinifolia > H. stipulacea > C. serrulata. Conclusion This antibacterial studies can further investigated on seagrasses for purification of bioactive substance and its possible utility in disease control.

898-901

Authors: Rengasamy Ragupathi Raja Kannan, Rajasekaran Arumugam, Perumal Anantharaman

Objective To search for new sources of safe and inexpensive antioxidants, the leaf, root and rhizome of Enhalus acoroides (E. acoroides) were screened for the first time for their antioxidant activities. Methods Samples of leaf, root and rhizome from E. acoroides were tested for total phenolic content, proanthocyanidins and in vitro antioxidant activity in terms of total antioxidant assay, DPPH assay and FRAP assay. Results The leaf sample was found to have high levels of phenolic (0.323±0.028 mg TAE/g) and proanthocyanidins (0.570 0±0.000 3 mg TAE/g) when compared to root and rhizome. The leaf samples exhibited higher total antioxidant activity (11.770±0.026 mg Ascorbic acid equivalent/g), higher percentage of DPPH radical scavenging activity (25.76±0.04) and higher reducing power (18.060±0.073) in terms of mg GAE/g. In addition, there was a significant correlation between total phenolic content and total antioxidant activity (R2=0.923), DPPH assay (R2=0.509), FRAP assay (R2=0.994). Similarly, significant correlation was found between the proanthocyanidin and total antioxidant activity (R2=0.977), DPPH assay (R2=0.464) and the FRAP assay (R2=0.998). Conclusion These results suggested that E. acoroides have strong antioxidant potential. Further study is necessary for isolation and characterization of the active antioxidant agents, which can be used to treat various oxidative stress-related diseases.

906-912

Authors: Hamdan Ibrahim Al-Mohammed, Tarek Tawfik Amin, Elsayed Aboulmagd, Hatem Refaat Hablus, Burhan Omar Zaza

Objective To investigate the prevalence of different intestinal parasitic infections among male primary schoolchildren in Al-Ahsa, Saudi Arabia, and to explore the possible relative socio-economic, environmental and behavioural sanitary correlates. Methods A cross-sectional descriptive study was performed. A total of 1 289 male students aged from 7 to 12 years were selected randomly from 20 urban and 12 rural primary schools by multistage sampling method. Data collection was carried out by self administered questionnaire form to the parents/guardians of students and included inquires about socio-demographics, environmental conditions, and behavioural sanitary habits. Stool analysis was carried out to examine the presence of intestinal parasitic infections. Results Overall prevalence of parasitic infections was 27.2%, more among rural students. Frequently encountered infections included Entamoeba histolytica/dispar (8.2%), Giardia lamblia (6.5%), Entamoeba coli (4.0%) and Enterobius vermicularis (1.6%). Logistic regression revealed that lower maternal educational level and occupational status, low family income, big family size, poor personal hygienic practices and positive history of previous intestinal infections among family members increased the likelihood of infections. Conclusion Although of low magnitude, intestinal parasitic infections still represent a public health concern among male schoolchildren in Al-Ahsa region. Socio-demographic and poor personal hygienic habits are the main predictors for these infections.

917-921

Authors: GTA Jombo, SK Gyoh

Cutaneous leishmaniasis is regarded as a re-emerging disease due to its increase in spread and rate of transmission over the past decade due to a proportionate increase in global human movements. This study was therefore carried out to review epidemiology, laboratory diagnosis and treatment with in-depth discussion on some of the rare clinical presentations of the disease capable of influencing its control. The study was based on literature review on clinical and laboratory features of cutaneous leishmaniasis from original research articles, review articles, short communications, letters to editor and case reports on the disease for the past 10 years (April 2000 to April 2010). The results were analysed using simple descriptive methods. The rarest presentations of leishmaniasis encountered were peritibial ulcérations, hard painful nipple in a male, swollen upper lip, dermatofibro sarcoma protuberans, sternal proliferative growth, turban tumour, post operative granuloma, chalazion-like 0.00%, and cutaneous sterile pyogranuloma with asymmetrical alopecia 0.00% each. Others include infected sebaceous cysts 0.45%, syphilis 0.19%, yaws 0.13%, thrombotic ulcérations 0.40%, mycetoma 0.90%, sarcoidosis 0.67%, painless nasal solitary nodule 0.00%, tuberculous lymphadenopathy 0.25% and unilateral erythema nodosum 0.28%. Physicians practicing in cutaneous leishmaniasis endemic, but most especially in the non-endemic areas should bear in mind that the disease may not be that rare in the context of the present global village phenomenon; and that, the next case could just be that of cutaneous leishmaniasis, the clinical picture of the patient notwithstanding.

762-764

Authors: Yingzi Lin, Shixiang Bao, Guanghong Tan, Fengying Huang, Huiqin Huang

Objective To construct and identify recombinant plasmid p UIS3-BLC+. Methods The cDNA of B-lymphocyte chemoattractant (BLC) was amplified from the total RNA of spleen tissues by PCR method, and were inserted into plasmid of Plasmodium berghei with UIS3 knockout by digestion of restrictive endonuclease and T7 ligation. The recombinant plasmids were screened, and then underwent restriction enzymatic digestion and DNA sequencing. Then the confirmed plasmid was further transfected into COS-1 cells by lipofectamine and the BLC expression was tested by RT-PCR and Western blotting. Results The cDNA of BLC gene was correctively amplified by RT-PCR and the recombinant plasmid p UIS3-BLC+ was constructed successfully, which was confirmed by restriction enzymatic digestion and DNA sequencing. RT-PCR and Western blotting also showed the BLC gene expression in COS-1 cells. Conclusions The recombinant plasmid p UIS3-BLC+ has BLC expression in COS-1 cells, and is useful for further study on BLC transgene and UIS3 gene knockout in Plasmodium berghei.

769-774

Authors: Subramanion Jo Thy Lachumy, Sreenivasan Sasidharan, Vello Sumathy, Zakaria Zuraini

Objective To elucidate its pharmacological activities and medicinal potential of extract of Etlingera elatior (E. elatior). Methods Phytochemical screening of the flower extract was done to determine the phytochemical in the extract. The pharmacological study included the determination of antimicrobial activity and minimum inhibitory concentration (MIC) of metabolic flower extract. The antimicrobial activity of the extract was tested against medically important bacterial, yeast and fungal strains. Apart from that, the methanolic extract of E. elatior flower was further tested in vivo toxicity using the brine shrimp lethality test. Moreover, the flower extract was qualitatively screened for their free radical scavenging activity by 2, 2-diphenyl-1-picrylhydrazyl radical (DPPH) assay. Results The extract was effective on tested microorganisms and MIC values were in the range of 1.563-50.000 mg/mL. The brine shrimp lethality test exhibited no significant toxicity (LC50 = 2.52 mg/mL) against Artemia salina. The E. elatior flower extract with high LC50 value signified that this plant is not toxic to humans. While the phytochemical screening of the flower extract revealed the presence of the following compounds: flavonoids, terpenoids, saponin, tannins and carbohydrates whereas, alkaloids, anthraquinone and reducing sugars were absent. The concentration of the flower extract required for 50% inhibition of DPPH radical scavenging effect (IC50) were 9.14 mg/mL and 8.08 mg/mL for butylated hydroxytoluene 8.08 mg/mL. Conclusions These findings indicate that the extract of E. elatior flower possesses pharmacological properties and potential to develop natural products based pharmaceuticals products.

783-787

Authors: Meena Kumari, Eesha BR, Mohanbabu Amberkar, Sarath babu, Rajshekar, Neelesh Kumar

Objective To evaluate the wound healing effect of aqueous extract of Crotalaria verrucosa (C. verrucosa) in rats. Methods Three wound models including incision, excision and dead space wounds were used in this study. The parameters studied were breaking strength in incision models, granulation tissue dry weight, breaking strength and hydroxyproline content in dead space wounds, percentage of wound contraction and period of epithelialization in excision wound model. Results Two doses of the extract with and without dexamethasone showed significant increases in mean hydroxyproline, total protein content and dry weight of granulation tissue but it was higher with dose 800 mg/kg comparing with the control. The dexamethasone treated group showed a significant (P<0.001) reduction in the wound breaking strength when compared to control group in incision type of wound model. Coadministration of C. verrucosa with dexamethasone significantly (P<0.001) increased the breaking strength compared to the dexamethasone treated only group. In excision wound model, the percentage of the wound contraction was significantly (P<0.01) increased by two doses of test extract on all the days except the lower dose which exhibited only on 12 th, 16 th days of drug treatment and it also reversed the dexamethasone suppressed wound contraction. It significantly (P <0.001) reduced the time required for epithelialization and reversed the epithelialization delaying effect of dexamethasone (P<0.001). Conclusions C. verrucosa was found to possess significant wound healing property. This was evident by decrease in the period of epithelialization, increase in the rate of wound contraction, skin breaking strength, and granulation tissue dry weight content. Hence C. verrucosa could be a good wound healing agent.

791-793

Authors: Sarmistha Saha, Gagan Goswami

Objective To investigate the gastroprotective activity of hydroalcoholic extract leaves of Ficus religiosa (F. religiosa) in different experimental models of gastric ulcer in rats. Methods The hydroalcoholic extract leaves of F. religiosa were studied at two dose levels (250 and 500 mg/kg, oral) in rats against absolute ethanol (0.2 mL oral), aspirin (200 mg/kg) and pyloric ligation induced gastric ulcer. Ranitidine (50 mg/kg, oral) was used as a standard drug. Mean ulcer indices and oxidative stress were measured. Phytochemical tests and acute toxicity tests were also carried out. Results Administration of F. religiosa to rats significantly decreased the ulcer index value when compared with the control treated group. Ranitidine (50 mg/kg, oral) also produced a significant decrease the ulcer index value when compared with the control treated group. Phytochemical analysis revealed the presence of tannins, sterols, saponins, flavonoids, carbohydrates and proteins. Conclusions The results suggest that the leaves of the F. religiosa possess significant anti ulcer activity.

797-799

Authors: D Devina Merin, S Prakash, B Valentine Bhimba

Objective To explore the biosynthesis of silver nanoparticles synthesized by marine microalgae. Methods Marine microalgae was collected from Central Marine Fisheries Research Institute (CMFRI, tuticorin) and cultured in the lab. Silver nanoparticles synthesis were observed in normal and microwave irradiated microalgae and screened against human pathogens for the presence of antimicrobials. Results The presence of silver nanoparticle was confirmed by UV-Visible spectroscopy at 420 nm by the presence of plasmon peak. Further confirmation was done by scanning electron microscope (SEM). Conclusions These results not only provide a base for further research but are useful for drug development in the present and future.

804-806

Authors: Hosseini Vasoukolaei Nasibeh, Telmadarraiy Zakkyeh, Vatandoost Hassan, Yaghoobi Ershadi Mohammad Reza, Hosseini Vasoukolaei Morteza, Oshaghi Mohammad Ali

Objective To determine the tick species parasitizing domestic ruminants in Ghaemshahr county in Mazandaran, a Caspian province in the north of Iran. Methods About 361 sheep, 54 goats and 10 cattle of 18 herds in several villages in Ghaemshahr were inspected for tick infestation. Separated ticks were preserved in 70% alcohol and identified. Results About 323 ticks (207 female, 116 male) were collected, the occurrence of ticks on sheep, goats and cattle were 28.3%, 22.2% and 20.0% respectively. The mean number of ticks on each animal was low (3-5 ticks per animal). Rhipicephalus sanguineus, Rhipicephalus bursa, Ixodes ricinus, Boophilus annulatus, Haemaphysalis punctata and Haemaphysalis numidiana were the tick species we found. Rhipicephalus sanguineus were the most abundant species in the study area. The largest number of ticks were generally present from April to July, mostly in animal ears and tails. Ixodes, Boophilus and Haemaphysalis occurred in mountainous areas of Ghaemshahr, whereas Rhipicephalus were present in both mountains and plains of the study area. Conclusions The result of this study is a survey of tick species from domestic animals in Iran and implication of possible prevention measures for diseases transmitted by ticks.

812-814

Authors: Chintana Chirathaworn, Yong Poovorawan, Viboonsak Vuthitanachot

Objective To determine the levels of IL-18 and IL-18 binding protein (BP) in patients with dengue virus infection. Methods Acute and convalescent sera were collected from each patient. Control group was sera from blood donors. The levels of both IL-18 and IL-18BP were measured by ELISA assays. Results It was shown that IL-18 and IL-18BP levels were significantly higher in patients when compared with controls. In addition, the level of IL-18BP was lower in convalescent than in acute sera. Conclusions These data suggest that both IL-18 and IL-18BP production was induced following dengue virus infection. Investigating the regulation of IL-18 by its natural regulator could lead to further understanding of the immune response or immunopathogenesis following dengue virus infection.

818-820

Authors: Vishwanath Sathyanarayanan, Ragini Bekur, Abdul Razak, Joydeep Chakraborty

Objective To study disseminated cryptococcal infection in a tertiary care hospital in Southern India. Methods The clinical profile of 12 disseminated cryptococcosis patients with the age group of 28-52 years was retrospectively analyzed. Results 7(58.3%) presented with fever < 30 days and 3(25%) > 30 days whereas 2(16.7%) did not have fever. All the 12(100%) had headache, 2(16.7%) had altered sensorium, one (8%) seizure. 5(41.7%) had diarrhea and vomiting. 6(50%) had oral candidiasis, and anemia. 9(75%) had elevated erythrocyte sedimentation rate (ESR). 6(50%) had neck stiffness. Cerebrospinal fluid (CSF) pressure was elevated in all 12(100%) patients. Blood culture positive for Cryptococcus neoformans(C. neoformans) in 11(91.7%) and CSF culture positive in all 12 (100%), one (8%) had urine culture positive. India ink preparation was positive in 10(83.3%). CD4 count was less than 50/microl in 4 (33.3%), between 50-100 in 6(50%) and 2(16.7%) in the range of 100-200. 6(50%) were treated with parenteral amphotericin B (0.7 mg/kg/d) during intensive phase followed by oral fluconazole 400 mg/d for 8 weeks then maintenance oral fluconazole 200 mg/d. 5(41.6%) were treated with fluconazole alone. 8(66.7%) improved and 4(33.3%) patients died. Among those who succumbed to the illness, 2(16.7%) received amphotericin and fluconazole, 2(16.7%) patients received fluconazole alone. Conclusions Disseminated cryptococcosis can cause considerable mortality in HIV patients and immunocompromised non- HIV individuals. At times, its presentation closely mimics that of Tuberculosis. Early diagnosis and appropriate treatment should be started as early as possible.

828-832

Authors: Ezeani Michael Chukwudi, Onyenekwe CC, Wachukwu CK, Anyiam DCD, Meludu SC, Ukibe RN, Ifeanyichukwu M, Onochie A, Anahalu I, Okafor UU

Objective To investigate the prevalence of microbial antigenic components of circulating immune complexes amongst grades of CD4 T lymphocyte counts in HIV sero positive and sero-negative participants. Methods Polyethelene glycol (PEG-600) and buffering methods of precipitation and dissociation of immune complexes was used to generate immune solution from sera of 100 HIV sero-positive and 100 HIV sero-negative participants. These were categorized into 3 grades based on CD4 count: > 500 cell/mm3, 200-499 cell/mm3 and <200 cell/mm3. The immune solutions were assayed using membrane based immunoassay and antibody titration, along side its unprocessed serum for detection of various microbial antigens and or antibodies. CD4 T cell counts were estimated using Patec Cyflow SL-3 Germany. Results Antigenic component of immune complexes of various infectious agents was detected in 99 and 70 HIV sero-positive and HIV sero-negative participants, respectively. In group A, there were 10 HIV positive participants, including 4 (40.0%) had circulating immune complexes(CIC s) due to Salmonella species only; 1(10.0%) due to Salmonella-Plasmodium falciparum (P. falciparum), Salmonella-P. falciparum-HCV and P. falciparum antigens, respectively. In group B, 45(45.4%) HIV sero-positive participants with CIC s had CD4 T lymphocyte count between 200-499 cells/mm3. Out of these, 20(44.4%) had CIC s due to Salmonella species only; 9(20%) due to Salmonella-P. falciparum. In group C, there were 44(44.4%) HIV sero-positive participants, including 3(6.8%) due to Salmonella species only; 24(54.4%) due to Salmonella-P. falciparum; 2(4.5%) due to P. falciparum only. Conclusions In HIV sero-positive participants, presence of heterogeneity of Salmonella species-P. falciparum antigens was highly incriminated in CD4 count depletion but not homogeneity of malaria parasites antigens. Malaria parasites antigens only were incriminated in CD4+ count depletion amongst HIV sero-negative participants. Before taking any decision on the management of HIV-1-positive individuals, their malaria and Salmonella paratyphi status should be assessed, but not malaria status alone.

835-837

Authors: Vishwanath Sathyanarayanan, Bekur Ragini, Abdul Razak, M Mukhya prana Prabhu

Neurobrucellosis is a rare form of localized brucellosis usually with no systemic manifestations. We report a rare case of brucellosis presenting as meningoencephalitis associated with hydrocephalus. This patient had a lymphocytic predominant CSF and was initially treated with empirical anti tubercular therapy and steroids. A week later, when his CSF culture grew Brucella species, the treatment was changed to a combination of streptomycin, doxycycline and rifampicin and the patient improved with this therapy. This case illustrates the need to consider neurobrucellosis as a close differential diagnosis of neurotuberculosis in endemic areas when the patient presents with meningo encephalitis with lymphocytic CSF

678-681

Authors: Haripriya Duraiswamy, Selvan Nallaiyan, Jeyakumar Nelson, Periasamy Rathina Samy, Marimuthu Johnson, Irudayaraj Varaprasadam

Objective To investigate the antimicrobial activity of some commonly available herbs against poultry pathogens such as Escherichia coli (E. coli) and Pseudomonas species isolated from poultry litter. Methods The extracts of Selaginella involvens and Selaginella inaequalifolia were tested against E. coli and Pseudomonas isolated from poultry litter by the agar diffusion method. Results Results indicated that different plant extracts showed inhibitory effects against E. coli (8-13 mm) and Pseudomonas (6.5-13 mm). The four different extracts of Selaginella involvens and Selaginella inaequalifolia showed similar levels of antimicrobial activity on E. coli. Conclusions The antimicrobial activities of all the four plant extracts are comparable and their potential as alternatives in the treatment of infections by these microorganisms were present in the poultry litter. Susceptibility testing is conduced on isolates using drugs selected on the basis of their importance to human medicine and use in poultry production.

687-690

Authors: Shalini Adiga, Prabhav Trivedi, V Ravichandra, Debashree Deb, Forum Mehta

Objective To evaluate potential memory enhancing effect of Punica granatum peel extract on rats. Methods Healthy adult male albino rats of Wistar strain were used. Each group of 6 rats were administered either distilled water or 50 mg/kg of extract or 100 mg/kg of extract for 15 days and subjected to passive avoidance test or T-maze test. In the next phase rats were administered distilled water or 100 mg/kg of extract for 15 days and the rats were given injection diazepam before subjecting them to the tests. Results The overall performance was better in test groups compared to control groups. Among the test groups, 100 mg/kg rats performed better than 50 mg/kg. The effect on spatial learning parameters like mean number of alternations and mean percentage bias was more marked compared to retention testing parameters like latency. 100 mg/kg Punica extract treated group also improved performance of diazepam treated rats. Conclusions There is a definite trend of memory improvement by Punica granatum peel with effects being more marked on spatial learning tendency and long term memory than on retention capacity.

696-701

Authors: Rajamani Karthikeyan, ST Somasundaram, T Manivasagam, T Balasubramanian, P Anantharaman

Objective To investigate the protective effects of brown alga Padina boergesenii (P. boergeseii) against carbon tetrachloride (CCl 4) induced oxidative damage and liver fibrosis in rats. Methods To assess the hepatic damage liver weight, the activities of TBARS level, glutathione, SOD, CAT and GP x in circulation and liver. Results The group of rats induced with CCl 4 alone (2 mL/kg body weight), showed noticeable increase in the liver weight and TBARS level. Followed by, the level of antioxidant enzymes glutathione peroxidase, Superoxide dismutase and catalase was also significantly (P <0.01) diminished. Where as, the rats pretreated with P. boergesenii (150 mg/kg body weight) modulated the CCl 4 induced liver fibrosis. The level of antioxidant enzymes and lipid peroxidation products was found to be significantly (P <0.01) attenuated to near normal level, when compared with rats induced with CCl 4 alone. In order to assess the role of carotenoids in the relevant activity, total carotenoid content of the extract was analysed and found to be 0.59 mg / g fresh weight (FW). Further, the histopathogical studies provide a supportive evidence for this study to show the protective nature of P. boergesenii. Conclusions The protective role of brown alga P. boergesenii extract has confirmed its potential activity through its antioxidant sparing actions against CCl 4 i nduced free radical damage. However, the possible mechanism of hepatoprotection is rather speculative at this stage and investigations are underway to isolate and characterize the bioactive compounds from P. boergesenii.

707-710

Authors: Jasminder Kaur, Xavier Rathinam, Marimuthu Kasi, Khoo Miew Leng, Rajasekaran Ayyalu, S Kathiresan, Sreeramanan Subramaniam

Objective To evaluate the antibacterial activity of the methanolic extract of mango (Mangifera indica L.) seed kernel. Methods Chokanan mango seed kernel and seed kernels from assorted mango varieties were collected, cleaned, dried and powered. Crude methanolic extracts of mango seed kernel were analyzed for the phytochemical constituents. The free radical scavenging activity was determined by 2,2-Diphenyl-1-picrylhydrazyl (DPPH) assay. Antibacterial activity was evaluated by disc diffusion assay with three medically important bacterial pathogens such as methicillin resistant Staphylococcus aureus (S. aureus) (MRSA), Escherichia coli (E. coli) and Vibrio vulnificus (V. vulnificus). Results Qualitative phytochemical analysis indicated the presence of important phytochemical compounds such as glycosides, saponins, flavanoids, tannins and alkaloids. There was no significant difference in the phytochemical content between the single and assorted mango seed kernels. However, the free radical scavenging study indicated that the assorted mango kernels showed slightly higher activity than the single species (P <0.05). The crude methanolic extract of mango seed kernel at a concentration of 100 mg/mL is found to have potential antimicrobial activity against MRSA and E. coli compared to V. vulnificus. Study on the antibacterial activity also indicated that there was no significant difference in the antibacterial activity of the single and assorted mango seed kernel extracts. Conclusions The present study conclusively demonstrates the free radical scavenging activity and antibacterial activities of mango seed kernel. In addition, the results also indicated that there is no significant difference in the phytochemical content and biological activity of mango kernels from single and assorted mango varieties.

715-722

Authors: Vishal Chaturvedi, Amere Subbarao Sreedhar

Objective To examine the role of heat shock protein 90 (Hsp 90) in the maintenance of actin cytoskeleton in human neuroblastoma tumor cells. Methods Co-precipitation experiments were performed to examine Hsp 90 interaction with actin. Hsp 90 and actin interactions were evaluated by protein refolding and acto-myosin motility assays. 17-(Allylamino)-17-demethoxygeldanamycin (17AAG) induced actin-cytoskeleton re-organization was examined by laser scanning confocal microcopy. Results It was shown that inhibition of Hsp 90 by 17AAG accelerates detergent induced cell lysis of neuroblastoma tumor cells through destabilization of actin cytoskeleton. The in vitro co-precipitation experiments showed that functional but not mutant Hsp 90 binds with F-actin. Among biochemical modifications, phopshorylation and oligomerization enhanced Hsp 90 binding with F-actin. F-actin binding to Hsp 90 interfered with Hsp 90 chaperone activity in protein refolding assays, and Hsp 90 binding to F-actin interfered with actin motility on myosin coated flow cell. In the combination treatment, 17AAG irreversibly augmented the effect of cytochalasin D, an inhibitor of actin polymerization. Conclusions It can be concluded that Hsp 90 binds to F-actin in tumor cells and maintains the cellular integrity. The results display a novel element of Hsp 90 inhibition in destabilizing the actin cytoskeleton of tumor cells, therefore suggest that 17AAG combination with cytoskeletal disruptor may be effective in combating cancer.

727-730

Authors: Manas Paramanik, Goutam Chandra

Objective To collect information on the seasonal activity of filarial vector around foothills of Susunia of West Bengal, India. Methods Indoor-resting mosquitoes were collected from the human habitations of 16 villages. Collected mosquitoes were identified and examined for different parameters following standard methods recommended by World Health Organization and pioneer workers. Results Overall man hour density, infection rate, infectivity rate, human blood index and daily mortality rate of the vector were assessed as 10.54%, 6.31%, 1.38%, 77.33% and 13.00%, respectively, which were 12.17%, 8.63%, 2.26%, 100.00% and 17.00%, respectively in the summer season. Average load of microfilaria, 1 st stage, 2 nd stage and 3 rd stage larvae of Wuchereria bancrofti in infected vectors were 10.02, 7.50, 5.56 and 4.68, respectively, which were 11.58, 9.90, 6.27 and 5.44, respectively in summer. Among the searched shelters 63.37%, 8.16% and 2.08% were found to be invaded by any vector, infected vector and infective vector, respectively, which were 68.75%, 12.76% and 3.91%, respectively in summer. Conclusions Different indices related to the vector were much higher in summer, which indicates that, summer is the most favorable season for transmission of lymphatic filariasis in the study area. Available data will help to formulate an effective vector control measure.

734-737

Authors: Hossein Motamedi, Hadis Mirzabeigi, Tahere Shirali

Objective To determine the pattern of antibiotic resistance among Staphylococcus aureus (S. aureus) isolates from clinical specimens and to identify community-acquired methicillin-resistant Staphylococcus aureus (CA-MRSA) in specimens that have been collected from patients referring to one of the hospitals of Ahvaz. Methods S. aureus isolates from a hospital in Ahvaz were screened for resistance to various antibiotics including methicillin. The susceptibility of the isolates was determined by Kirby-Bauer disc diffusion method. The MRSA was also treated with ethidium bromide to find the origin of resistance. Results Among the bacterial isolates, all of 11 S. aureus were resistant to methicillin and cefixime, 2 were resistant to ciprofloxacine, 6 were resistant to tetracycline and the reminder were sensitive or intermediate to other antibiotics. The treated isolates were reminded resistant to methicillin and this suggested that the plasmid was not the origin of resistance in these isolates. Conclusions These results showed that infection due to MRSA is widespread in Ahvaz and with respect to the spread of vancomycin resistance among MRSA and appearance of overwhelming infections. It is necessary to identify continuously the profile of antibiotic resistance among S. aureus isolates in other regions and finding appropriate antibiotic for infection control and eradication.

741-744

Authors: MU Rasheed, Z Ahmed

Objective To evaluate the detection of methicillin resistant Staphylococcus aureus (MRSA) and analyze the performance of Mastalex MRSA (Mast, UK). Methods Two hundred and ten Staphylococcus aureus (S. aureus) strains were isolated from different clinical samples and were tested for methicillin resistance by Oxacillin (1 μg) and Cefoxitin (30 μg) disc diffusion, oxacillin agar screen, and minimum inhibitory concentration of oxacillin and cefoxitin. S. aureus isolates were grown on the blood agar and mannitol salt agar with (2 mg/L) and without oxacillin for the analysis of Mastalex MRSA. Results Out of 210 S. aureus strains tested, 103 strains were detected as methicillin resistant by Cefoxitin disk diffusion, Cefoxitin minimal inhibitory concentration (MIC) and Mastalex MRSA test. Whereas oxacillin disc diffusion and oxacillin agar screen detected 91 and 97 MRSA respectively. The Cefoxitin MIC test performance was equivalent to Cefoxitin disc diffusion. 103 (100%) strains grown on blood agar without and with oxacillin, and 76 (74%) and 93 (91%) strains grown on mannitol salt agar without and with oxacillin shown positive agglutination with Mastalex MRSA test respectively. Conclusions The cefoxitin disk diffusion/Mastalex MRSA is very suitable for detection of MRSA and the tests can be an alternative to PCR for detection of MRSA in resource constraint settings. Mastalex test would be particularly useful when confirmation of resistance is urgently required.

748-753

Authors: Jerapan Krungkrai, Waranya Imprasittichai, Sumintra Otjungreed, Sawirasagee Pongsabut, Sudaratana R Krungkrai

Malaria is a major cause of morbidity and mortality in the developing world. This situation is mainly due to emergence of resistance to most antimalarial drugs currently available. Artemisinin-based combination treatments are now first-line drugs for Plasmodium falciparum (P. falciparum) malaria. Artemisinin (qinghaosu) and its derivatives are the most rapid acting and efficacious antimalarial drugs. This review highlights most recent investigations into the emergence of artemisinin resistance in falciparum malaria patients on the Thai-Cambodian border, a historical epicenter for multidrug resistance spread spanning over 50 years. The study presents the first evidence that highlights the parasites reduced susceptibility to artemisinin treatment by prolonged parasite-clearance times, raising considerable concern on resistance development. Although the exact mechanism of action remains unresolved, development of resistance was proposed based from both in vitro experiments and human patients. Lines of evidence suggested that the parasites in the patients are in dormant forms, presumably tolerate to the drug pressure. The World Health Organization has launched for prevention and/or containment of the artemisinin-resistant malaria parasites. Taken together, the emergence of artemisinin resistance to the most potent antidote for falciparum malaria, poses a serious threat to global malaria control and prompts renewed efforts for urgent development of new antimalarial weapons.

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.