Asian Pacific Journal of Tropical Medicine

337-341

Authors: VU Omoja, AO Anaga, IR Obidike, TE Ihedioha, PU Umeakuana, LI Mhomga, IU Asuzu, SM Anika

Objective To investigate the effects of combination therapy of methanolic leaf extract of Azadirachta indica (A. indica) and diminazene diaceturate (DDA) in the treatment of experimental Trypanosoma brucei brucei (T. brucei brucei) infection in rats. Methods Acute toxicity study of the drug and extract combinations were done. Selection of the best drug and extract combinations was carried out using fifty four rats of both sexes separated into 9 groups. Three dose combinations were derived from selection of the best drug and extract combinations used for the final study viz: 7 mg/kg bw DDA plus 125 mg/kg bw extract (group B), 3.5 mg/kg bw DDA plus 250 mg/kg bw extract (group C), and 1.8 mg/kg bw DDA plus 500 mg/kg bw extract (group D). The final study had in addition to the three groups derived from the dose response study, four other groups viz: uninfected untreated negative control (group F), infected and treated with 3 000 mg/kg bw extract alone (group E), infected and treated with 7 mg/kg bw DDA alone (group A), and infected untreated positive control (group G). The parameters assessed were onset of parasitaemia (OP), level of parasitaemia (LOP), clearance of parasites post treatment (COPPT), relapse infection period (RIP), post infection survival period (PIST). Results There was no significant difference in OP between the groups (P < 0.05). One day post treatment, the mean LOP of groups A, B, and C were found to be significantly lower than that of group D which in turn was lower than that of group E and G respectively. The mean LOP of group E was significantly lower than group G two days post treatment and this trend continued throughout the experimental period. Mean COPPT of group D was significantly longer than that of groups A, C and B. There was no significant difference in the mean COPPT among groups B, C and A. The mean RIP of group D was significantly shorter than group C, and that of group C was significantly shorter than that of group A. There was no relapse of infection in group B. The PIST of group E did not differ significantly from group G. Conclusions This experiment stands to conclude that combination of 125 mg/kg bw extract and 7 mg/kg bw DDA is very effective in the treatment of trypanosomosis, caused by T. brucei. This combination therapy proved to be better than single therapy of DDA.

347-352

Authors: Sunil Kumar, Vipin Kumar, Om Prakash

Objective To investigate antidiabetic, hypolipidemic histopathological analysis of Dillenia indica (D. indica) methanolic leaves (DIME) extract in alloxan induced diabetic rat by administering oral doses (250 and 500 mg/kg body weight). Methods Blood glucose levels were measured using blood glucose test strips with elegance glucometer on weekly intervals till the end of study (i.e. 3 weeks). Other parameters e.g. liver profile, renal profile and total lipid levels were determined in normal and alloxan induced diabetic rats after oral administration of the extract for 21 days. Histopathological changes in diabetic rat organs (pancreas, liver and kidney) were also observed after extract treatment. Results Daily oral administration DIME (250 and 500 mg/kg body weight) and glibenclamide (10 mg/kg) showed beneficial effects on blood glucose level (P < 0.001) as well as improving kidney, liver functions and hyperlipidaemia due to diabetes. The extract treatment also showed to enhanced serum insulin level and body weight of diabetic rats as compared to diabetic control group. Furthermore, the extract has a favorable effect on the histopathological changes of the pancreas, liver and kidney in alloxan induced diabetes. Conclusions D. indica possess antidiabetic property as well improve body weight, liver profile, renal profile and total lipid levels. DIME has also favorable effect to inhibit the histopathological changes of the pancreas and kidney in alloxan induced diabetes.

357-362

Authors: MJ Mendki, AK Sharma, Vijay Veer, OP Agrawal, Shri Prakash, BD Parashar

Objective To characterize the genetic structure of various populations of Culex quinquefasciatus (Cx. quinquefasciatus) from India representing different geoclimatic locations. Methods Inter simple sequence repeat (ISSR) markers were used. A set of 20 primers were screened with the laboratory populations of mosquito species. Finally the IS 40 primer was chosen based on the scorable banding pattern showing 100 percent polymorphism among the various populations. The statistical analysis was done using POPGENE 1.31 software. The consensus tree was generated based on UPGMA modified from NEIGHBOR procedure of PHYLIP Version 3.5. Results The cluster analysis shows the main cluster which is divided into two sub cluster representing all the populations separated as per their phylogeographic and geoclimatic condition. Conclusions The findings will be helpful in understanding the population variation under different ecological conditions and development of effective vector management strategies.

367-370

Authors: Vinod Kumar, Jalaj Kumar Gour, Surabhi Bajpai, Manish Mishra, Rakesh Kumar Singh

Objective To detect leishmanial antigens in pre and post treated urine of visceral leishmaniasis (VL) patients. Methods Urine and serum sample from three VL patients were collected. Ammonium sulphate precipitation and purification of urine sample was done for proteins isolation. SDS PAGE of proteins was done followed by western blotting, with the patient's pre and post treatment serum. Results Eight proteins of molecular weights 17 kDa, 25 kDa, 28 kDa, 42 kDa, 47 kDa, 54 kDa, 60 kDa and 85 kDa were detected in the urine of VL patients before treatment. After treatment with miltefosine, none of the above proteins was detected in urine samples. The western blot analysis with pre treatment serum confirmed the antigenicity of four urinary proteins of molecular weights 25 kDa, 28 kDa, 54 kDa and 60 kDa. The seropositivity with 25 kDa and 28 kDa antigens was negative with serum obtained after the completion of treatment. Conclusions In the context to unavailability of a prognostic tool, urinary leishmanial antigens may offer a better choice and may also be useful as immunoprophylactic candidates.

375-378

Authors: Rajendran Darling Anpin Raja, Solomon Jeeva, Juststella Wilfred Prakash, Johnson Marimuthu Antonisamy, Varaprasadham Irudayaraj

Objective To screen the antimicrobial potential of three ethnomedicinal plants Chassalia curviflora Thw. (C. curviflora), Cyclea peltata Hook. F. & Thomson (C. peltata) and Euphorbia hirta L (E. hirta) used in folk medicines in Aarukani hills Kani tribe, Tamil Nadu, India against human bacterial pathogens. Methods Antibacterial efficacy was performed by disc diffusion method against the pathogens viz., Escherichia coli (E. coli) (ATCC 35218), Staphylococcus aureus (S. aureus) (ATCC 6538), Salmonella typhi (S. typhi) (MTCC 733), Proteus vulgaris (P. vulgaris), Proteus mirabilis (P. mirabilis) and Streptococcus pyogenes (S. pyogenes) and incubated for 24 h at 37 °C. Results The maximum degree of antibacterial activity was observed in C. peltata followed by C. curviflora. While E. hirta showed comparatively low degree of antibacterial activity. The methanolic extract of C. peltata showed the antibacterial activity against three pathogens viz., S. pyogenes, P. vulgaris and E. coli with the inhibition zones 12 mm, 10 mm and 9 mm, respectively. hexane extracts of C. peltata also showed the antibacterial activity against two selected pathogens viz., P. vulgaris and P. mirabilis with 15 mm and 12 mm of inhibition zones. All the three different concentrations (0.25, 0.50 & 0.75 mg/mL) of methanolic extract of C. peltata show the inhibitory effect on the three susceptible bacteria S. pyogenes, P. vulgaris and E. coli with the maximum inhibition in the highest concentration (0.75 mg/mL). The methanolic and hexane extracts of C. curviflora exhibited the antibacterial activity against only one bacterium each i.e. P. vulgaris and S. typhi with the maximum zone of inhibition 13 and 11 mm respectively. The methanolic and hexane extracts of E. hirta exhibited the antibacterial activity against only one bacterium i.e. S. pyogenes with the maximum zone of inhibition 13 and 11 mm respectively. Conclusions The present investigation revealed that the C. curviflora, C. peltata and E. hirta are potentially good source of antibacterial agents and demonstrates the importance of such plants in traditional medicines.

386-390

Authors: Abu Arra Basma, Zuraini Zakaria, Lacimanan Yoga Latha, Sreenivasan Sasidharan

Objective To assess antioxidant activities of different parts of Euphorbia hirta (E. hirta), and to search for new sources of safe and inexpensive antioxidants. Methods Samples of leaves, stems, flowers and roots from E. hirta were tested for total phenolic content, and flavonoids content and in vitro antioxidant activity by diphenyl-1-picrylhydrazyl (DPPH) assay and reducing power was measured using cyanoferrate method. Results The leaves extract exhibited a maximum DPPH scavenging activity of (72.96±0.78)% followed by the flowers, roots and stems whose scavenging activities were (52.45±0.66)%, (48.59±0.97)%, and (44.42±0.94)%, respectively. The standard butylated hydroxytoluene (BHT) was (75.13±0.75)%. The IC50 for leaves, flowers, roots, stems and BHT were 0.803, 0.972, 0.989, 1.358 and 0.794 mg/mL, respectively. The reducing power of the leaves extract was comparable with that of ascorbic acid and found to be dose dependent. Leaves extract had the highest total phenolic content [(206.17±1.95) mg GAE/g], followed by flowers, roots and stems extracts which were (117.08±3.10) mg GAE/g, (83.15±1.19) mg GAE/g, and (65.70±1.72) mg GAE/g, respectively. On the other hand, total flavonoids content also from leave had the highest value [(37.970±0.003) mg CEQ/g], followed by flowers, roots and stems extracts which were (35.200±0.002) mg CEQ/g, (24.350±0.006) mg CEQ/g, and (24.120±0.004) mg CEQ/g, respectively. HPTLC bioautography analysis of phenolic and antioxidant substance revealed phenolic compounds. Phytochemical screening of E. hirta leaf extract revealed the presence of reducing sugars, terpenoids, alkaloids, steroids, tannins, flavanoids and phenolic compounds. Conclusions These results suggeste that E. hirta 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.

397-400

Authors: Vishwanath Sathyanarayanan, Abdul Razak, Kavitha Saravu, Shastry Barkur Ananthakrishna, M Mukhyprana Prabhu, KE Vandana

Objective To highlight the spectrum of clinical manifestations, labs, complications, treatment and outcome of brucellosis. Methods Retrospective study was conducted in Kasturba Medical College, Manipal University, Karnataka, India which included 68 confirmed cases of brucellosis from January 2006- April 2010. Diagnosis of brucellosis was made by culturing the sera/body fluids by standard BACTEC method (or) by testing the sera for Brucella agglutinins using the standard agglutination test (SAT). A titer of 1:320 or more was considered as significant. SPSS 16 was used for statistical analysis and Microsoft Excel for graphical representation. Results Of the 68 patients, 46 (68%) were male and 22 (32%) were female patients with age distribution of 9–75 years. Forty four (64.7%) had history of contact with unpasteurized dairy products or infected animals. Symptoms included fever (68, 100%), myalgia (21, 31%), musculoskeletal symptoms (23, 34%), headache (16, 24%), gastrointestinal symptoms (19, 28%) and altered sensorium (3, 4%). Co-morbidities and associations included HIV positivity (2, 3%), type 2 diabetes mellitus (13, 19%), steroid therapy (3, 4%) and HBsAg positivity (8, 12%). Ten (15%) patients had cervical lymphadenopathy, 4(6%) had splenic enlargement, 6 (7%) had hepatomegaly, 19 (28%) had hepatosplenomegaly and 2(3%) got meningeal signs. Anaemia was observed in 39 (57.3%) cases, high erythrocyte sedimentation rate (ESR) was present in 55 (80.8%) cases, leucocytosis in 10(14.7%), leucopenia in 10(14.7%), thrombocytopenia in 23 (33.82%) and thrombocytosis in 2 (2.94%) cases. Conclusions In countries like India, where brucellosis and tuberculosis are endemic; rapid, sensitive and highly specific diagnostic methods are required to make early diagnosis and prevent resistance as there is an overlap in therapy.

404-408

Authors: Adam H Al-Shamiri, Mohammed A Al-Taj, Ameera S Ahmed

Objective To determine the disease prevalence and its relationship with hepatitis B and C viruses among school children in five endemic areas by schistosomiasis. Methods During June 2007 and March 2009, 1 484 school children aged between 5 – 16 years participated in the current study from 32 basic schools in five districts (Al-Dhabab, Hedran, Warazan, Al-Barh and Al-Shmaytin) in Taiz Governorate, Republic of Yemen. Out of school children who participated in the study; 1 406 stool samples, 1 484 urine samples and 214 blood samples were collected and examined. Results Schistosoma mansoni (S. mansoni) was found in all the studied areas except Al-Barh. However, Schistosoma haematobium (S. haematobium) was recorded only in Al-Shmaytin and Al-Barh. Both S. mansoni and S. haematobium were observed in Al-Shmaytin district. The overall prevalence was 20.76% for S. mansoni and 7.41% for S. haematobium. The prevalence rate of infection among males was higher than females, shwing no significant differences. Rate of light, moderate and heavy infections in the case of S. mansoni were 41.78%, 25.34% and 32.87% respectively. Whereas, for S. hematobium it was 50.90% for light infection and 49.09% for heavy infection. Regarding to the prevalence of viral hepatitis among infected school children with schistosomiasis, it could be noticed that hepatitis B virus was higher than the prevalence of hepatitis C virus. But, the presence of HBsAg and anti-HCV was not associated with Schistosoma infection. Conclusions Schistosomiasis infection is an important public health problem in Taiz Governorate, Republic of Yemen. There was a correlation between S. haematobium and hepatitis B, but no association between S. mansoni infections and hepatitis B and C viruses.

412-413

Authors: Jaydeep Sarkar, Charu Mohan, DN Misra, Atul Goel

The spectrum of dengue infection varies from dengue fever to more severe forms such as dengue hemorrhagic fever (DHF) and dengue shock syndrome (DSS). In DHF bleeding may be overt (external) or internal. At time bleeding can occur at unusual sites. We report one such case of DHF in which bleeding occurred within the tongue manifesting as an intra-lingual hematoma, causing significant upper airway obstruction.

417-420

Authors: Neelam Kaistha, Varsha Gupta, Shailpreet Sidhu, Jagdish Chander

Salmonella are most commonly associated with gastroenteritis and enteric fever in humans. Occasionally, dissemination of bacilli throughout the body results in establishment of localized foci of persistent infection especially in patients with debilitating diseases and immunosuppressive states. Infection at various aberrant sites due to Salmonella has been reported relatively seldom. It has perfected the art of intracellular survival in niches from where they can cause myriad of effects. Six cases with Salmonella infection at unusual sites without any preexisting or underlying disease diagnosed over a period of two years are presented here. Salmonella etiology was not suspected in these patients and the diagnosis was made microbiologically only after culture isolation.

253-258

Authors: Anthony A Attama, Collins N Igbonekwu

Objective To formulate and evaluate in vitro, surface-modified solid lipid microspheres containing halofantrine using lipid matrix formed from goat fat and a phospholipid (P90H). Methods The model drug, halofantrine in an increasing concentration of 1%, 2%, 3%, 4% and 5% w/w was incorporated into surface-modified solid lipid microspheres formulated by hot homogenization. Effect of drug concentration on the encapsulation efficiency was studied. The dispersion was evaluated using particle size, particle morphology, pH and encapsulation efficiency. The drug formulation with highest encapsulation efficiency was selected and used for the release studies and compared with the release from a commercial dosage form (Halfan® 250 mg tablet, Glaxo-Smithkline, Mayenne France) using simulated gastric fluid (SGF pH 1.2), simulated intestinal fluid (SIF pH 7.2) and phosphate buffer (pH 6.8) as biorelevant media. Results were analyzed statistically and the level of significance was taken to be P < 0.05). Results Discrete and spherical solid lipid microspheres were produced. The particle size of the dispersion was low (32.48–33.87 μm) with minimal particle growth and high encapsulation efficiencies (86.8%–91.0%) after 3 months. The pH of the microspheres dispersion changed appreciably after 3 months. In vitro release result obtained revealed sustained and controlled drug release from the lipid microspheres compared with the tablet dosage form. Conclusions Formulation of halofantrine as solid lipid microspheres presents a better alternative to the conventional tablet formulation as the in vitro dissolution of the highly lipophilic halofantrine was highly improved.

266-270

Authors: Qiang Wu, Peng Zhou, Shiyun Qian, Xi Qin, Zhigang Fan, Qiongyao Fu, Zhinong Zhan, Hua Pei

Objective To clone and express Rv3265c gene of Mycobacterium tuberculosis in Escherichia coli (E. coli) under optimistic conditions, obtain and identify protein expressed, analyze the structure and characteristics of the protein using bioinformatics methods for future applications. Methods Rv3265c gene from Mycobacterium tuberculosis H37Rv was amplified by polymerase chain reaction, and was cloned into the pET-30a vector after purification and recovery. The recombinant plasmid was sequenced and expressed in E. coli BL21(DE3), and then purified and identified by western blotting. The essential physical-chemical properties of the protein were predicated by bioinformatics tools, including subcellular location, secondary structure, domains, antigenic epitopes, etc. Tertiary structure of the protein based on homology modeling was established, while multi-sequence homological alignment and phylogenetic analysis were proformed. Results The recombinant protein was obtained in soluble fraction from expression system in E. coli BL21(DE3) carrying pET30- Rv3265c plasmid, and Rv3265c gene was expressed correctly. Bioinformatics analysis showed the protein contained no signal peptide and transmembrane helices, located outside of membrane. Secondary structure analysis revealed it contained α-helix, extended strand and random coil, 46.8%, 14.6%, 38.6%, respectively. Furthermore, it possessed six potential antigenic epitopes, one glycosyl transferase domain. A simple three-dimensional model of this protein was constructed by Swiss-model sever. Both sequences and structures were conservative and especial either in gene or in protein. Conclusions Rv3265c gene might be a desirable molecular target for anti-tuberculosis drug and vaccine. The purified protein from expression will be utilized to study the kinetics of L-rhamnosyltransferase and to develope an enzyme assay for screening vaccine or drug.

275-277

Authors: Ximin Hu, Jianwei Du, Chongjin Tong, Shanqing Wang, Jian Liu, Yuchun Li, Changhua He

Objective To understand the current epidemiological status of Angiostrongylus cantonensis in Hainan island and provide the scientific evidences for prevention and control of the disease. Methods The investigation on nature infection condition of intermediate hosts and final hosts of Angiostrongylus cantonensis were carried out in five counties, Hainan Province, and enzyme linked immunosorbent assay (ELISA) was employed to detect antibodies of Angiostrongylus cantonensis in human hosts. Results A total of 1612 molluscs were examined, 21.3% of which harbored L3 of Angiostrongylus cantonensis. Among them, the infection rates of Pomacea canaliculata, Achatina fulica, Cepaea and Phlegm bilineatus were 12.36%(64/518), 22.66%(121/534), 20.93%(9/43), and 28.24%(146/517), respectively. Of the 118 rats trapped, 13 Rattus norvegicus were found to be infected with Angiostrongylus cantonensis. A total of 459 serum samples were collected and tested. 92 serum samples were Angiostrongylus cantonensis antibody-positive. Conclusions The survey revealed a wide distribution of Angiostrongylus cantonensis in Hainan island. Pomacea canaliculata and Achatina fulica are main intermediate hosts of Angiostrongylus cantonensis. Rattus novegicus is a nature definitive host, indicating that a considerable number of people are at risk of angiostrongyliasis. Health education, rigorous food inspection and surveillance are all needed to prevent angiostrongyliasis outbreaks in future.

283-287

Authors: Tolulope O Ajala, Cecilia I Igwilo, Ibrahim A Oreagba, Oluwatoyin A Odeku

Objective To investigate the antiplasmodial activity of the extracts of Phyllanthus amarus (P. amarus) on Plasmodium yoelii (P. yoelii) (a resistant malaria parasite strain used in animal studies) infection in mice. Methods The aqueous and ethanol extracts of the whole plant of Phyllanthus amarus was administered to Swiss albino mice at doses of 200 mg/kg/day, 400 mg/kg/day, 800 mg/kg/day and 1600 mg/kg/day and the prophylactic and chemotherapeutic effect of the extracts against P. yoelii infection in mice was investigated and compared with those of standard antimalaria drugs used in the treatment of malaria parasite infection. Acute toxicity test was carried out in mice to determine the safety of the plant extract when administered orally. Results The results showed that the extracts demonstrated a dose—dependent prophylactic and chemotherapeutic activity with the aqueous extracts showing slightly higher effect than the ethanol extract. The antiplasmodial effects of the extracts were comparable to the standard prophylactic and chemotherapeutic drugs used in chloroquine resistant Plasmodium infection although the activity depended on the dose of the extract administered. The extracts showed prophylactic effect by significantly delaying the onset of infection with the suppression of 79% at a dose of 1600 mg/kg/day. Conclusions The results obtained indicate that the extracts of the whole plant of P. amarus possess repository and chemotherapeutic effects against resistant strains of P. yoelii in Swiss albino mice. The findings justify the use of the extract of P. amarus in traditional medicine practice, for the treatment of malaria infections.

294-300

Authors: S Rajan, T Thirunalasundari, S Jeeva

Objective To evaluate the phytochemical and anti-bacterial efficacy of the seed kernel extract of Mangifera indica (M. indica) against the enteropathogen, Shigella dysenteriae (S. dysenteriae), isolated from the diarrhoeal stool specimens. Methods The preliminary phytochemical screening was performed by the standard methods as described by Harborne. Cold extraction method was employed to extract the bioactive compounds from mango seed kernel. Disc diffusion method was adopted to screen antibacterial activity. Minimum inhibitory concentration (MIC) was evaluated by agar dilution method. The crude extracts were partially purified by thin layer chromatography (TLC) and the fractions were analyzed by high performance thin layer chromatography (HPTLC) to identify the bioactive compounds. Results Phytochemical scrutiny of M. indica indicated the presence of phytochemical constituents such as alkaloids, gums, flavanoids, phenols, saponins, steroids, tannins and xanthoproteins. Antibacterial activity was observed in two crude extracts and various fractions viz. hexane, benzene, chloroform, methanol and water. MIC of methanol fraction was found to be (95±11.8) μg/mL. MIC of other fractions ranged from 130–380 μg/mL. Conclusions The present study confirmed that each crude extracts and fractions of M. indica have significant antimicrobial activity against the isolated pathogen S. dysenteriae. The antibacterial activity may be due to the phytochemical constituents of the mango seed kernel. The phytochemical tannin could be the reason for its antibacterial activity.

305-309

Authors: Subramanian Bragadeeswaran, Selvam Priyadharshini, Kolandhasamy Prabhu, Solomon Raj Sophia Rani

Objective To study the antimicrobial, hemolytic activity and immunomodulatory activity of fish epidermal mucus and their chemical constituents from Cynoglossus arel (C. arel) and Arius caelatus (A. caelatus). Mucus plays an important role in the prevention of colonization by parasites, bacteria and fungi. Methods Epidermal mucus was obtained from two marine fishes, lyophilized and the chemical composition of epidermal mucus was analysed by FT-IR analysis. The in vitro antimicrobial activity against human pathogens (fungi, gram positive and gram-negative bacteria) and also the hemolytic activity and immunomodulatory activity were determined. Results Totally ten human pathogens were tested against the fish mucus. Out of the ten pathogens, five pathogens have proved to be sensitive to the mucus. Maximum zone of inhibition was observed against Vibrio cholera (V. cholera) (9 mm and 2 mm in diameter), followed by Staphylococcus aureus (S. aureus) with a inhibition zone of (6 mm and 3 mm), Streptococcus areus (S. areus) (5 mm and 4 mm), Vibrio parahemolyticus (V. parahemolyticus) (4 mm and 5 mm) respectively. Conclusions The present investigation has revealed that positive progresses in the fish mucus extracts against human pathogens and hemolytic activity. But further efforts are required for the purification and isolation of the active antimicrobial compounds in order to establish their possible applications.

315-319

Authors: Iquo B Otu-Bassey, Monday F Useh, Anyanwu A Alaribe

Objective To investigate the relationship between enterobiasis and enuresis before and after albendazole therapy among 632 children, aged, 5–14 years, in Calabar, Nigeria. Methods The scotch tape (cellotape) technique was used for the detection of eggs of Enterobius vermicularis (E. vermicularis) while questionnaire-based interviews were used for screening for anal itching and/or enuresis among study participants. All subjects found positive for Enterobius infection as confirmed by the presence of eggs and those with persistent anal itching (both enuretic and non-enuretic) were treated with 400 mg of albendazole given as a single dose. The treated subjects were re-assessed post-treatment to ascertain whether they were cured and also to determine their enuretic status (for the enuretics). Results The overall prevalences of Enterobius infection, anal itching, and enuresis prior to albendazole therapy were 6.8%, 42.9%, and 35.6% respectively. There was no statistically significant difference in the prevalence of these parameters by the socio-economic class of subjects (P = 0.462, P = 0.647, and P = 0.610, respectively). The pre-treatment prevalences of enuresis among Enterobius and anal itching-positive subjects were 53.5% and 49.8%, respectively versus 34.3% and 24.9%, respectively among their negative counterpart (P = 0.012 and P < 0.001, respectively). Four (20%) out of the 20 enuretic children found Enterobius egg-negative after albendazole therapy were equally cured while 8(40%) had reduction of their enuresis, thus giving a total resolution rate of 60%. Similarly, 64.2% resolution (25% cure and 32.8% reduction) of enuresis was observed amongst 120 anal itching-cured/enuretic children. Enuretic status of the Enterobius-/anal itching-uncured subjects, on the other hand, remained unchanged post-treatment. Conclusions This study is suggestive of the involvement of E. vermicularis in the aetiology of enuresis in Calabar. Children presenting with, especially, uncomplicated enuresis should be screened for enterobiasis.

323-324

Authors: Rajat Jhamb, Bineeta Kashyap, GS Ranga, A Kumar

Dengue fever (DF) and dengue haemorrhagic fever (DHF) are important mosquito-borne viral diseases of humans and recognized as important emerging infectious diseases in the tropics and subtropics. Compared to nine reporting countries in the 1950s, today the geographic distribution includes more than 100 countries worldwide. Dengue viral infections are known to present a diverse clinical spectrum, ranging from asymptomatic illness to fatal dengue shock syndrome. Mild hepatic dysfunction in dengue haemorrhagic fever is usual. However, its presentation as acute liver failure (ALF) is unusual. We report a patient with dengue shock syndrome who presented with acute liver failure and hepatic encephalopathy in a recent outbreak of dengue fever in Delhi, India.

328-329

Authors: Shamma Arora, Naveen Gupta, Ashwani Kumar, IR Kaur

Non typhoidal Salmonella species are thought to be potentially infectious to humans. We isolated Salmonella enteritidis from a 10-year-old boy with fever and thrombocytopenia. We reviewed the literature concerning infections caused by Salmonella but we could not find any such case report from India.

169-175

Authors: Nisar Ahmad, Hina Fazal, Bilal H Abbasi, Mazhar Iqbal

Objective To investigate in vitro larvicidal and antioxidant enzymes potential of the medicinal plants Ginkgo biloba (G. biloba), Stevia rebaudiana (S. rebaudiana) and Parthenium hysterophorous (P. hysterophorous) against Anopheles stephensi (An. stephensi) 4th instars larvae. Methods For evaluation of larvicidal potential, the ethanolic, methanolic and dichloromethane leaves extracts of three different plants were used in dose-dependent experiments in two media, while the antioxidant enzymes activities were investigated using four different methods viz., superoxide dismutase, peroxidase, ascorbate and catalase. Results An. stephensi has developed resistance to various synthetic insecticides, making its control increasingly difficult. The comparative performance of ethanolic extracts (65%-90%) was found better than the methanolic extract (70%-87%) and dichloromethane extract (60%-70%). Among the three plants extracts tested in two media, S. rebaudiana exhibited higher larvicidal activity with LC50 (24 h) in methanolic extract than P. hysterophorous and G. biloba. G. biloba and P. hysterophorous exhibited the strongest antioxidative enzymes activity and S. rebaudiana were less active and no significant difference was observed. Conclusions These three plants exhibit larvicidal potential and can be further used for vector control alternative to synthetic insecticide due to eco-friendly and diseases control, furthermore these plant species have potent antioxidative enzyme activities, therefore, making them strong natural candidate particularly for diseases which are caused due to free radicals.

182-184

Authors: OL Erukainure, JA Ajiboye, RO Adejobi, OY Okafor, SB Kosoko, FO Owolabi

Objective To investigate the ability of the methanolic extract of pineapple peel to attenuate alcohol-induced changes in total phospholipids and lipid peroxidation in brain tissues. Methods Oxidative stress was induced by oral administration of ethanol (20% w/v) at a dosage of 5 mL/kg bw in rats. After 28 days of treatment, the rats were fasted overnight and sacrificed by cervical dislocation. Brain tissues were assayed for total phospholipid (TP) content and malondialdehyde (MDA). Results Administration of alcohol significantly caused a reduction in TP content. Treatment with pineapple peel extract significantly increased the TP content. Significant high levels of MDA was observed in alcohol-fed rats, treatment with pineapple peel extract significantly reduced the MDA levels. Conclusions Results obtained from this study indicates that pineapple peel extract protects against alcohol-induced changes in total phospholipids and lipid peroxidation in brain tissues.

192-195

Authors: G Madhumitha, AM Saral

Objective To investigate the phytochemical, antibacterial, antifungal and anticandidal activity of successive extracts of Crossandra infundibuliformis (Acanthaceae) leaves. Methods Preliminary screening on the presence of alkaloids, saponins, phytosterols, phenolic compounds, flavanoids, tannins, carbohydrates, terpenoids, oils and fats were carried out by phytochemical analysis. The antibacterial, antifungal and anticandidal activities were done by agar well diffusion technique. Results The successive extracts have an array of chemical constituents and the MIC values of antibacterial activity ranges from 0.007 8 to 0.015 0 μg/mL. In case of antifungal and anticandidal activities the MIC values were between 0.125 and 0.250 μg/mL. Conclusions These findings demonstrate that the leaf extracts of C. infundibuliformis presents excellent antimicrobial activities and thus have great potential as a source for natural health care products.

201-204

Authors: Jackson Rafael Oliveira Peixoto, Giselle Cristina Silva, Renata Albuquerque Costa, Joseí res Lira de Sousa Fontenelle, Gustavo Hitzschky Fernandes Vieira, Antonio Adauto Fonteles Filho, Regine Helena Silva dos Fernandes Vieira

Objective To evaluate the antibacterial effect of aqueous and ethanolic moringa leaf extracts (Moringa oleifera) on the growth of gram-positive and negative bacteria. Methods Paper disks were soaked with 100, 200, 300 and 400 μL of extract at 20 g/180 mL and 10 g/190 mL. All extracts were tested against Escherichia coli (ATCC25922), Staphylococcus aureus (ATCC25923), Vibrio parahaemolyticus, Enterococcus faecalis (ATCC29212), Pseudomonas aeruginosa (ATCC27853), Salmonella enteritidis (IH) and Aeromonas caviae. The susceptibility tests were performed using the modified disk diffusion method. Results The strains E. coli, P. aeruginosa and S. enteritidis (IH) were resistant to all treatments. In general, disks with 400 μL extract were the most efficient against S. aureus, V. parahaemolyticus, E. faecalis and A. caviae. Conclusions The study indicates a promising potential for aqueous and ethanolic Moringa leaf extracts as alternative treatment of infections caused by the tested strains.

212-214

Authors: Cheng Lan, Xiaoning Sun, Liwei Dong, Baili Huang, Su Yuan, Keli Wu

Objective To study the role and the mechanism of endotoxin in the pathogenesis of gastric mucosa during portal vein hypertension gastrography (PHG) in the rats with cirrhosis. Methods Rat model for PHG was established by injection of tetrachloride. The animals were injected with endotoxin i.p. at 3 mg/kg and endotoxin antagonist BPI21 i.v. at 2.0 mg/kg. The plasma level of endotoxin as well as the gastric mucosal level of tumor necrosis factor alpha (TNF-α) was measured with azobenzene and ELISA respectively. Furthermore, the pathological changes of the gastric mucosa were studied with HE stainning. Results In rats with PHG, increased endotoxin and TNF-α as well as the gastric pathological lesion were observed. Injection of endotoxin remarkably increased plasma level of endotoxin as well as the gastric mucosal level of tumor necrosis TNF-α and induced more serious gastric lesion. Animals injected with endotoxin antagonist BPI21 showed improved gastric mucosal lesion, accompanied by the declining TNF-α level. Conclusions Our results suggestes that endotoxin may play a pathogenetic role in PHG by inducing the expression of TNF-α.

220-223

Authors: Mehdi Khoobdel, Behroz Davari

Objective To determine the faunal diversity of Muscidae and Fanniidae flies in Tehran, Iran. Methods A net-capturing by direct observing method and fly trap were used to capture adult flies. To determine the fauna of flies in different habitats, 4 biotopes including corpse (human, birds, livestock), garbage and decaying organic matters, animal carcasses and human indoor habitat were selected. Big hashing nets (95 cm in diameter) have been used for adult flies capture in these biotopes. Results In this study, totally 2 418 adult flies from 8 families including Muscidae, Fanniidae, Calliphoridae, Sarcophagidae, Tachinidae, Syrphidae, Anthomyiidae and Conopidae were captured. Among these, 1 279 belong to Muscidae and Fanniidae families. Four genera and 5 medically important species captured from two above mentioned families. Conclusions Based on scientific documentation, 2 species of Muscina stabulans (M. stabulans) and Fannia scalaris (F. scalaris) are reported for the first time in Iran. However M. stabulans is a cosmopolitan species and its presence in Iran was probable.

227-228

Authors: A Mariana, S Vellayan, I Halimaton, TM Ho

Objective To identify the acari present on pet Burmese pythons in Malaysia and to determine whether there is any potential public health risk related to handling of the snakes. Methods Two sub-adult Burmese pythons kept as pets for a period of about 6 to 7 months by different owners, were brought to an exotic animal practice for treatment. On a complete medical examination, some ticks and mites (acari) were detected beneath the dorsal and ventral scales along body length of the snakes. Ticks were directly identified and mites were mounted prior to identification. Results A total of 12 ticks represented by 3 males, 2 females and 7 nymphal stages of Rhipicephalus sanguineus (R. sanguineus) were extracted from the first python while the other one was with 25 female Ophionyssus natricis (O. natricis) mesostigmatid mites. Only adult female mites were found. These mites are common ectoparasites of Burmese pythons. Conclusions Both the acarine species found on the Burmese pythons are known vectors of pathogens. This is the first record that R. sanguineus has been reported from a pet Burmese python in Malaysia.

234-240

Authors: Shiva Raj Adhikari, Siripen Supakankunti, M Mahmud Khan

Objective To examine the choice of healthcare providers for treating kala azar (KA) in Nepal. Methods Information was collected from clinically diagnosed KA patients seeking care from public hospitals located in KA endemic districts. The survey collected information from more than 25 percent of total KA cases in the country. For empirical estimation of probability of choosing a provider-type as a first contact healthcare provider, a multinomial logit model was defined with five alternative options with self care as the reference category. Results The empirical model found that price of medical care services, income of households, knowledge of patients on KA and KA treatment, borrowing money, age of patient, perceived quality of provider types, etc. determine the likelihood of seeking care from the alternative options considered in the analysis. All variables have expected signs and are consistent with earlier studies. The price and income elasticity were found to be very high indicating that poorer households are very sensitive to price and income changes, even for a severe disease like KA. Using the empirical models, we have analyzed two policy instruments: demand side financing and interventions to improve the knowledge index about KA. Conclusions Due to high price elasticity of KA care and high spillover effects of KA on the society, policy makers may consider demand side financing as an instrument to encourage utilization of public hospitals.

248-250

Authors: LJ Zhang, GQ Wang, HL Yu, J Wang, SW Wang, Y Jia, YY Yu, JG Xu

We isolated a novel strain of Alphaproteobacteria from a patient, who had medical history of chronic rhinitis for more than twenty years and recently experienced local skin abscess and ulcer. He eventually died of multiple organ failure due to multi-antibiotics resistance. We identified the microorganism by 16SrRNA sequencing and found that it belonged to the genus Rhodoplanes. It was named as Rhodoplanes sp. strain ZLJ-0. It is resumed that Rhodoplanes sp. strain ZLJ-0 might be an emerging human pathogen involving in unknown febrile conditions and could cause local infection of any tissues or organs. Differential diagnosis of febrile patients should be conducted in clinical practice and research on emerging pathogens of Alphaproteobacteria should be performed to determine the epidemiology, clinical symptoms and pathogenic features of these pathogens.

85-87

Authors: Zhigang Fan, Lingmin Zhang, Guogang Yan, Qiang Wu, Xiufeng Gan, Saifeng Zhong, Guifen Lin

Objective To analyse the structure and function of NADPH-cytochrome p450 reductase (CYPOR or CPR) from Plasmodium falciparum (Pf), and to predict its' drug target and vaccine target. Methods The structure, function, drug target and vaccine target of CPR from Plasmodium falciparum were analyzed and predicted by bioinformatics methods. Results PfCPR, which was older CPR, had close relationship with the CPR from other Plasmodium species, but it was distant from its hosts, such as Homo sapiens and Anopheles. PfCPR was located in the cellular nucleus of Plasmodium falciparum. 335aa–352aa and 591aa – 608aa were inserted the interior side of the nuclear membrane, while 151aa–265aa was located in the nucleolus organizer regions. PfCPR had 40 function sites and 44 protein–protein binding sites in amino acid sequence. The teriary structure of 1aa–700aa was forcep–shaped with wings. 15 segments of PfCPR had no homology with Homo sapien CPR and most were exposed on the surface of the protein. These segments had 25 protein–protein binding sites. While 13 other segments all possessed function sites. Conclusions The evolution or genesis of Plasmodium falciparum is earlier than those of Homo sapiens. PfCPR is a possible resistance site of antimalarial drug and may involve immune evasion, which is associated with parasite of sporozoite in hepatocytes. PfCPR is unsuitable as vaccine target, but it has at least 13 ideal drug targets.

92-96

Authors: Shiv Kumar Verma, Vikas Kushwaha, Vijaya Dubey, Kirti Saxena, Aakanksha Sharma, Puvvada Kalpana Murthy

Objective To investigate which life stage of the parasite has the ability to stimulate release of pro- or anti-inflammatory mediators from macrophages. Methods The human macrophage/monocyte cell line THP-1, the mouse macrophage cell line RAW 264.7 and naive peritoneal macrophages (PM) from the rodent host Mastomys coucha (M. coucha) were incubated at 37 °C in 5% CO2 atmosphere with extracts of microfilariae (Mf), third stage infective larvae (L3) and adult worms (Ad) of Brugia malayi. After 48 hr post exposure, IL-1β, IL-6, TNF-α, IL-10 and nitric oxide (NO) in cell-free supernatants were estimated. Results Extracts of all the life stages of the parasite were capable of stimulating pro- (IL-1β, IL-6 and TNF-α) and anti-inflammatory (IL-10) cytokines in both the cell lines and peritoneal macrophages of M. coucha. Mf was the strongest stimulator of pro-inflammatory cytokines followed by L3 and Ad; however, Ad was a strong stimulator of IL-10 release. Mf was found to have potential to modulate LPS-induced NO release in RAW cells. Ad-induced NO release was concentration dependent with maximum at 20 μg/mL in both RAW and PMs. Conclusions The results show that parasites at all life stages were capable of stimulating pro- (IL-1β, IL-6 and TNF-α) and anti-inflammatory (IL-10) cytokines and NO release from macrophages of susceptible host M. coucha, human and mouse macrophage cell lines. Mf can suppress the LPS-induced NO release in RAW cells. The findings also show that the two cell lines may provide a convenient in vitro system for assaying parasite-induced inflammatory mediator release.

101-105

Authors: Mohammed Shahid, Anuradha Singh, Farrukh Sobia, Mohammad Rashid, Abida Malik, Indu Shukla, Haris Manzoor Khan

Objective To delineate the frequency of occurrence of blaCTX-M, blaTEM, and blaSHV in Enterobacteriaceae from North-Indian tertiary hospital. Methods A random collection of a subset of 45 Escherichia coli (E. coli) and 28 Klebsiella pneumoniae (K. pneumoniae) that was resistant to a third generation cephalosporin and obtained during 2007–2008 was selected for detailed screening for blaCTX-M, blaTEM, and blaSHV by monoplex PCRs. The isolates demonstrating the presence of blaCTX-M alleles were characterized for the specific CTX-M-genogroup by using a multiplex PCR. Results Resistance to cefoperazone, ceftazidime, ceftriaxone, cefotaxime, cefoxitin and piperacillin was 100% each in K. pneumoniae isolates, whereas these resistance-rates for E. coli isolates were 93.1%, 83.8%, 91.9%, 93.6%, 97.3% and 97.1%, respectively. Concomitant resistance to aminoglycosides, quinolones and aztreonam was also noticed. Presence of any of the bla genes (blaCTX-M, blaTEM, and blaSHV) was noticed in a total of 28 (38.4%) isolates of the 73 isolates studied. Many isolates demonstrated occurrence of these genes in various combinations. blaCTX-M, blaTEM, and blaSHV were noticed in 28.8%, 10.9% and 13.7% isolates, respectively. Multiplex PCR in blaCTX-M harboring isolates demonstrated the presence of CTX-M-Genogroup-1 and sequencing for the specific CTX-M-type revealed presence of CTX-M-15 type. RAPD typing showed wide diversity in isolates. Conclusions This is amongst the premier report describing the simultaneous occurrence of blaTEM, blaSHV, and blaampC in Indian Enterobacteriaceae and that wider dissemination of these genes, as demonstrated by diversity of isolates, raises concern and emphasizes a need for extensive search for the presence of these gene pools in Indian subcontinent.

112-116

Authors: Madhu SK, Vijayan VA, Shaukath AK

Objective To isolate the larvicidal component from the fruits of Piper longum (P. longum) against the filariasis vector, Culex quinquefasciatus (C. quinquefasciatus). Methods Pulverized fruits of P. longum were subjected to soxhlet extraction using series of organic solvents of increasing polarity. All the solvent extracts were verified for their larvicidal efficacy against 4th instar larvae of C. quinquefasciatus employing standard WHO procedure. Bioassay-guided fractionation through column chromatography lead to the isolation of a bioactive amide, pipyahyine from the petroleum ether extract. Results Petroleum ether extract was found to be the most active fraction among all the extracts tested with LC50 and LC90 being 1.03 and 2.04 ppm respectively. Whereas, pipyahyine, an isolated component of the same fraction was found to be even more effective than the parent extract in terms of LC50 being 0.58 and 1.88 ppm respectively. Conclusions From the results, it is evident that P. longum can be considered as a powerful arsenal for the control of mosquito population.

121-124

Authors: Uche Fidelia Ijeoma, Shorinwa Olusayo Aderonke, Okorie Ogbonna, Mgbahurike Amaka Augustina, Chijioke-Nwauche Ifeyinwa

Objective To investigate antinociceptive and anti-inflammatory activities of crude extract from Ipomoea involucrata leaves (Convolvulaceae) in mice and rats. Methods The antinociceptive activity was tested using acetic acid-induced abdominal writhing test in mice. The anti-inflammatory activity was evaluated using egg albumin induced oedema of rat paw. Results Phytochemical screening showed the presence of alkaloids, flavonoids, saponins, terpenoids and tannin. At the doses of 25–100 mg/kg, Ipomoea involucrata exhibited dose-dependent and significant increase in pain threshold in acetic acid –induced writhing test of mice (P<0.05, student t-test) The administration of Ipomoea involucrata leaf extract (25–100 mg/kg) showed dose-dependent decreases in paw volume of egg albumin induced oedema in rats and a significant higher anti-inflammatory activity compared to the standard control (Aspirin). Conclusions These results support the claims on the traditional use of the of Ipomoea involucrata leaves in the treatment of toothache, rheumatic pains and other inflammatory conditions. Studies on the isolation and structural elucidation of the active principle are still needed being carried out.

129-132

Authors: Koffi Koudouvo, Simplice D. Karou, Denise P. Ilboudo, Kouami Kokou, Kodjo Essien, Kodjo Aklikokou, Comlan de Souza, Jacques Simpore, Mensavi Gbéassor

Objective To investigate the antimalarial effect of a few plants in Togo folk medicine. Methods After ethnobotanical survey, Opilia celtidifolia, Pavetta corymbosa (P. corymbosa) and Tamarindus indica (T. indica) were selected for screening. In vitro antimalarial tests were performed on crude extracts against fresh clinical isolates of Plasmodium falciparum using the semi microtest. Results Different IC50 values of the extracts ranged from 2.042 to 100.000 μg/mL. According to the results, the methanol extract of aerial part of P. corymbosa followed by aqueous extract of fruit of T. indica were the most active (IC50 of 2.042 and 4.786 μg/mL, respectively). Qualitative test revealed the presence of alkaloids in the leaves of P. corymbosa that may be responsible for the activity of the plant. Conclusions Our study provides scientific evidence for usage of plant in the folk medicine, and further studies are needed for identification and purification of the active principles.

137-139

Authors: Ishaleku D, Sar TT, Houmsou RS

Objective To evaluate the prevalence of Sabmonella typhi (S.typhi) among subjects attending College of Education Health Clinic, Akwanga, Nasarawa state from the year. 2005 to 2007 and to recommend some preventive measures to the populace. Methods Blood samples were tested for infection using the widal test. Results Out of the 793 patients examined, 579(73.0%) were positive with 174 (30.05%), 254(43.86%) and 151(26.07%) for the years 2005, 2006 and 2007 respectively. The age range with the highest prevalence of infection for the period was 21–30 with 207(35.75%) followed by 31–40 year group with 133 (22.97%). Chi-square analysis showed no significant difference in infection between males and females (P>0.05). Conclusions The results of this study provide a useful guide in the formulation of S. typhi control measures in tertiary institutions in the State and also help to check the spread of the pathogen in the general population.

146-147

Authors: Ibrahim EA, Kheir MM, Elhardello OA, Almahi WA, Ali NI, Elbashir MI, Ishag Adam

Objective To investigate the levels of serum cortisol in patients with uncomplicated Plasmodium falciparum (P. falciparum) malaria in an area of unstable malaria transmission in eastern Sudan. Methods The concentrations of cortisol were measured in sera of 25 patients with uncomplicated P. falciparum malaria (at presentation, 24 h and 7 d later) and 25 healthy volunteers using radioimmunoassay gamma counter. Results There was no significant difference in mean (SD) of total cortisol levels in patients with malaria in comparison with the control group; 602.2 (369.6) vs. 449.2(311.7) ng/mL, P=0.12. In patients with uncomplicated P. falciparum malaria, the mean (SD) presenting cortisol levels were significantly higher in comparison to the levels on day 7; 602.2 (369.6) vs. 373.6(139.1) ng/mL, P=0.009. In the patients with uncomplicated P. falciparum malaria (on presentation) cortisol levels were not correlated with initial temperature or the presenting parasitaemia. Conclusions Thus, cortisol levels were not significantly different between the patients and the controls.

152-155

Authors: Igbiks Tamuno

Objective To investigate the prevalence of use of traditional medicines amongst patients with HIV infection receiving therapies of antiretroviral (ARV) drugs at the Aminu Kano Teaching Hospital (AKTH), Kano, Northwest Nigeria, and to assess the attitude of these patients to their ARV therapy. Methods A cross sectional prospective study using pretested structured questionnaires administered on 430 patients with antiretroviral therapy attending the AKTH between April and June 2009. Data was collected on socio-demographic characteristics, use of traditional medicine and attitude to antiretroviral therapy. Results A mean age of (33.6#20381;8.4) years old was found with 67.2% females and 32.8% males. A total of 29% had no formal education while 10.5% had postgraduate education; 12% earned above 35 000 naira (230 USD) per month; 63.8% were married; 39.8% had at least 2 sexual partners; 27.5% used traditional medicine before commencement of antiretroviral therapy (ART), but only 4.25% of patients used ARV and traditional medicine concurrently. There was no significant difference in most of the socio-demographic indices between the concurrent users and other patients (P>0.05). A total of 28.8% HIV patients, 14.6% patients used traditional medicine before ART and 29.4% concurrent users had missed at least a dose of their ARVs since commencement of therapy. 148 (37%) of the patients had their drug regimen changed at least once while 23 (20.90%) patients receiving traditional medicine before ART and 5 (29.41%) patients having two treatments had their drug regimen changed. Conclusions A total of 4.25% patients used ARV and traditional medicine concurrently. In conclusion, the widespread use of traditional medicine by patients living with HIV/AIDS should be of concern to clinicians and policy makers.

159-162

Authors: Yildirim Cesaretli, Ozcan Ozkan

Objective To classify and characterize spider bites among inquiries to the National Poison Information Center (NPIC) between 1995 and 2004, in terms of the epidemiology and clinical symptomatology. Methods Clinical and epidemiological data were obtained from the NPIC's patient records. The following information was recorded for each spider bite: demographics, circumstances of the bite, and local and systemic effects. Results A total of 82 cases were reported. The accidents were mostly seen during August. The gender distribution was 59.76% male, 37.20% female, and 2.44% unknown and the 20–29 age group presented more spider bites. Most of the cases were in the Central Anatolia, Marmara, Mediterranean, and Black Sea regions. Local symptoms were observed in 60.87% of the cases, including local pain, edema, redness, itching, debris, burning, and numbness. Systemic symptoms were observed such as nausea, vomiting, abdominal pain, lethargy, anxiety, weakness, somnolence, dyspnea, hypertension, hypotension, and hyperthermia. Conclusions In conclusion, these findings emphasize the presence of medically important spider species in Turkey. All patients and especially pediatric patients should be admitted to the hospital. Identification of spider species may be considered a useful clinical and epidemiological tool in determining the incidence and risk of spider bites.

167-168

Authors: Kiran Yagain, Mary Mathew

A two year old boy from southwestern village of India presented with a 2 cm × 1 cm size swelling in scrotum for 8 months. It was excised surgically and the histopathological examination revealed a gravid filarial worm. The occurrence of adult filarial worm in 2 year old boy signifies the endemicity and high larval load of insect vectors in western coast of south India.

925-930

Authors: Giuliana Giribaldi, Elena Valente, Amina Khadjavi, Manuela Polimeni, Mauro Prato

Objective To investigate the role of macrophage inflammatory protein-1alpha (MIP-1alpha) in the detrimental enhancement of matrix metalloproteinase-9 (MMP-9) expression, release and activity induced by phagocytosis of malarial pigment (haemozoin, HZ) in human monocytes. Methods Human adherent monocytes were unfed/fed with native HZ for 2 h. After 24 hours, MIP-1alpha production was evaluated by ELISA in cell supernatants. Alternatively, HZ-unfed/fed monocytes were treated in presence/absence of anti-human MIP-1alpha blocking antibodies or recombinant human MIP-1alpha for 15 h (RNA studies) or 24 h (protein studies); therefore, MMP-9 mRNA expression was evaluated in cell lysates by Real Time RT-PCR, whereas proMMP-9 and active MMP-9 protein release were measured in cell supernatants by Western blotting and gelatin zymography. Results Phagocytosis of HZ by human monocytes increased production of MIP-1 alpha, mRNA expression of MMP-9 and protein release of proMMP-9 and active MMP-9. All the HZ-enhancing effects on MMP-9 were abrogated by anti-human MIP-1alpha blocking antibodies and mimicked by recombinant human MIP-1alpha. Conclusions The present work suggests a role for MIP-1alpha in the HZ-dependent enhancement of MMP-9 expression, release and activity observed in human monocytes, highlighting new detrimental effects of HZ-triggered proinflammatory response by phagocytic cells in falciparum malaria.

937-940

Authors: Khalid M Bindayna, Mariam Murtadha

Objective To determine the molecular epidemiology of extended-spectrum β-lactamase (ESBL) by testing a cohort of clinical ESBL-producing bacterial isolates that were isolated in the Kingdom of Bahrain. Methods ESBL producing Enterobacteriaceae isolates (based on phenotypic tests) were collected from Microbiology Laboratory of the Salmaniya Medical Complex, Bahrain between January-June 2006. Antibiotic susceptibility to a panel of antibiotics was performed and blaCTX-M genes were detected by multiplex PCR. Results A total of 230 isolates (Escherichia coli, n=180; Klebsiella pneumoniae, n=50) were studied, 98% were CTX-M type. For Escherichia coli isolates, 65 (36.1%) harbored CTXM+TEM combination and 68 (37.8%) had CTX-M alone. In contrast, for Klebsiella pneumoniae isolates only 5 (10.0%) harbored the CTX-M combination, and none had CTX-M only. The blaCTX-M gene was found predominantly in urine isolates (n=145/230; 63.0%). Sensitivity to imipenem and nitrofurantoin was 100% and 60%, respectively. CTX-M carriage was associated with the resistance to fluoroquinolones, trimethoprim-sulfamethoxazole and aminoglycosides. Conclusions Our study documentes high prevalence of CTX-M ESBL type among Escherichia coli and Klebsiella from the Kingdom of Bahrain. The apparent dissemination of CTX-M producers could represent a substantial barrier in the treatment of community-acquired infections. The use of extended-spectrum cephalosporins, quinolones, and aminoglycosides is compromised, leaving carbapenems as the therapeutic option for severe infections caused by ESBL producers.

948-951

Authors: Sankaran Rajkumar, Arulsamy Jebanesan, Rajarathinavelu Nagarajan

Objective To assess the larvicidal and egg hatching inhibition property of the leaf essential oil of Coccinia indica (C. indica) against Anopheles stephensi (An. stephensi). Methods The larvicidal potential of C. indica leaf essential oil was evaluated against 1st, 2nd, 3rd and 4th instars larvae of An. stephensi using WHO protocol. The 24h LC50 and LC90 values of the essential oil were determined following probit analysis. The egg hatching inhibition activity was also tested at 10, 20, 40, and 60 mg/L. The IC50 value of essential oil was determined against eggs of An. stephensi. Results The essential oil extracted from C. indica possessed excellent larvicidal and egg hatching inhibition activity against An. stephensi. The bioassays showed LC50-LC90 of 54.3-140.3, 65.5-155.6, 86.8-180.7 and 95.3-192.6 for 1st, 2nd, 3rd, and 4th larval instars, respectively. The 50% egg hatching inhibition concentration (IC50) was noted at 16.5 mg/L. Conclusions The present finding suggest that the C. indica leaf essential oil provided an excellent potential for controlling An. stephensi mosquito at earlier stage of their life cycle.

959-963

Authors: Nagaraj Selvaraj, Baskar Lakshmanan, Papiya Mitra Mazumder, Muthukumar Karuppasamy, Simon Santosh Jena, Ashok Kumar Pattnaik

Objective To evaluate the wound healing and antimicrobial activity of root extracts of Ixora coccinea (I. coccinea). Methods To investigate the wound healing efficacy of root extract of I. coccinea Linn, five groups of animals were divided each containing six animals. Two wound models including incision and excision wound models were used in this study. The parameters studied were tensile strength on incision wound model and in terms of wound contraction for excision wound model were compared with standard Nitrofurazone (NFZ) ointment (0.2% w/w). Six extracts (ethanol, aqueous, petroleum ether, benzene, chloroform and ethyl acetate) of I. coccinea were screened for in vitro growth inhibiting activity against different bacterial strains viz, Staphylococcus aureus, Bacillus pumilius, Enterococcus faecalis, Escherichia coli, Salmonella typhi and Pseudomonas aeruginosa and fungi Candida albicans and Aspergillus niger were compared with the standard drugs ciprofloxacin and chloramphenicol for antibacterial and griseofulvin for antifungal screening. The serial dilution and cup (or) well plate methods were used for the antimicrobial study and MIC was determined. Results The ethanolic extract showed significant (P<0.001) wound healing activity when compared to standard drug NFZ with respect to normal control group. Amongst all, ethanolic extract showed highly significant antibacterial activity against all bacterial strains used in this study when compared to standard. The aqueous extract showed moderate significant inhibition against all bacterial strains when compared to standard. All the extracts were shown negligible activity against the fungal strains used in this study. Conclusions The ethanolic root extract of I. coccinea showed pronounced wound healing and antibacterial activity. The probable reason to heal the wound was that the external application of the extract prevented the microbes to invade through the wound thus the protection of wound occurs against the infection of the various organisms.

969-972

Authors: Jude E Okokon, Paul A Nwafor, Kufre Noah

Objective To evaluate the kidney protective effect of ethanolic root extract of Croton zambesicus (C. zambesicus) against gentimicin-induced kidney injury in rats. Methods The root extract (27-81 mg/kg) was administered to rats for eight days with concurrent administration of gentimicin (100 mg/kg) daily for the same period of time. Protective effect of the extract was evaluated in serum levels of creatinine, urea, and uric acid as well as some ions like sodium, potassium and chloride. Histological examination of the kidneys from different treatment groups were also carried out. Results Administration of the root extract significantly reduced histopathological changes in the kidneys of the extract-treated rats especially in the rats treated with lower doses of the extract (27 and 54 mg/kg). The levels of serum urea and creatinine were also reduced significantly (P<0.01) at these doses with no observable effect on the levels of uric acid and ions. Conclusions The kidney – protective activity of this extract could be due to its antioxidant and free radical scavenging activities.

978-984

Authors: Gupta Sandeep, Ahirwar Dheeraj, Neeraj Kumar Sharma, Deenanath Jhade, Ahirwar Bharti

Objective To assess the effect of plumbagin-free alcohol extract (PFAE) of Plumbago zeylanica Linn. (Plumbaginaceae) (P. zeylanica) root, on female reproductive system and fertility of adult female wistar rats. Methods After the oral acute toxicity study, the PFAE was administered at two dose levels to perform the estrous cycle study, anti-implantation and abortifacient activity and hormonal analysis. However, the estrogenic/antiestrogenic activity was evaluated at only one most effective dose. Results LD50 cut-off was 5 000 mg/kg body weight. The extract exhibited significant anti-implantation and abortifacient activity at the tested dose levels (300 and 500 mg/kg, p.o.) (P<0.01). The extract dose-dependently decreased the levels of serum progesterone, follicle stimulating hormone and luteinizing hormone, while a dose-dependent increase was observed in the concentration of serum prolactin. The extract did not show any significant changes in structure and function of uterus when given alone, but when given along with ethinyl estradiol, it exhibited significant antiestrogenic activity in immature overiectomized female rats(P<0.001). Biochemical parameters in the serum/blood and haematological parameters did not show appreciable changes throughout and after the course of investigation. However, all the altered parameters returned to normalcy within 30 days following withdrawal of treatment. Conclusions All findings suggest that the antifertility activity of extract could possibly be through the changes in the implantation site, altered hormonal levels, prolonged estrous cycle and anti-estrogenic activity. Hence, the extract possesses reversible antifertility activity without adverse toxicity in female rats.

993-996

Authors: UM Chukwuocha, KN Eke

Objective To investigate the relationship between malaria parasite status and cholesterol level of 110 consenting subjects (55 patients and 55 controls) in parts of the Imo River Basin of Nigeria. Methods Giemsa staining was used for malaria parasite examination while Randox cholesterol kit was used for cholesterol level estimation. Results About 49 persons (90%) with malaria had low cholesterol (<180 mg/dL). Highest mean cholesterol levels were 274 mg/dL for study patients and 220 mg/dL for controls respectively; Lowest mean cholesterol levels were 168 mg/dL (patients) and 138 mg/dL (controls) respectively. Low cholesterol levels (<180 mg/dL) were found in patients (84%) and controls (6%) respectively. However, 16.4% of controls and 6% of patients had borderline cholesterol level (200-239 mg/dL). This study establishes a significant correlation (12.9%, P<0.01) between malaria parasite status and cholesterol level. Conclusions These findings imply that cholesterol level estimation may be a potential concurrent and valuable diagnosis for malaria status.

1001-1006

Authors: Manisha DebMandal, Shyamapada Mandal, Nishith Kumar Pal

Objective To investigate the prevalence of leptospirosis among patients from within and outside Kolkata, India, attending the Calcutta School of Tropical Medicine, for treatment during August 2002 to August 2008. Methods The leptospirosis cases were determined on the basis of clinical, epidemiological, and biochemical factors, and were tested for leptospiral antibodies using IgM ELISA. Serum samples with absorbance ratio ≥ 1.21 were interpreted as reactive. Results The commonest presentation involved fever, headache and jaundice. The male-female ratio was 61:46. A total of 65(64.20%) cases had abnormal liver and renal functions respectively, and 57.1% had both the abnormalities. The highest incidence (75, 35.04%) was recorded in September-October followed by July-August (53, 24.77%). The reactive cases had absorbance ratios between 1.21 and 8.21, and 53 showed equivocal result, while IgM non reactivity were seen in 90 patients (absorbance ratios 0.10-0.90). The patients responded to treatment with parenteral antibiotics, penicillin, ceftriaxone and cefotaxime; follow up did not reveal case fatality. Conclusions The cardinal signs of leptospirosis help in making clinical diagnosis, but in any hyper-endemic situation any patient reporting with acute fever and signs of pulmonary, hepatic or renal involvement should be suspected to have leptospirosis and investigated accordingly. Increased awareness, and early diagnosis and treatment, can reduce mortality due to leptospirosis.