Asian Pacific Journal of Tropical Medicine

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.

510-514

Authors: Xueli Zheng, Jialin Hu, Santhosh Puthiya Kunnon, Chen Xiaoguang

Objective To identify necrophagous fly speies from different regions in China using inter simple sequenc repeat (ISSR) and sequence-characterized amplified region (SCAR) melocular markers and to analyze their gene difference and genetic relationship. Methods Five carrion fly species were collected from 12 cities and regions in China, including Musca domestica (M. domestica), Lucilia sericata (L. sericata), Chrysomya megacephala (C. megacephala), Helicophagella melanura (H. melanura), Boethcherisca peregrina, and they were studied using ISSR and SCAR markers. Results Eight ISSR primers were used for amplification of 121 samples. 679 clear and stable bands were identified, of which 516 bands were polymorphic. Several species-specific ISSR fragment were cloned and sequenced as an initial effort to derive the SCAR markers. Using M. domestica SCAR specific primers, SCAR-PCR amplification was performed for 8 M. domestca population sample DNA from different regions in China as well as L. sericata, C. megacephala, H. melanura and Lucillia cupirina. The result showed only M. domestica produced specificalty 600 bp fragment, but L. sericata, C. megacephala, H. melanura and Lucillia cupirina did not produce the same specific fragment. Clustering analysis showed clustering of most flies of M. domestica, C. megacephala and L. sericata. M. domestica samples from different regions in China yielded different banding patterns. Conclusions Application of ISSR-PCR and SCAR markers to identify necrophagous fly species from 12 cities and regions in China is first reported. ISSR-PCR and SCAR markers provide a quick reliable molecular marker technique for the identification of different species of necrophagous fly.

519-522

Authors: S De, D Pan, AK Bera, V Sreevatsava, SK Das, S Das, T Rana, S Bandyopadhyay, D Bhattacharya

Objective To study the canine echinococcosis by coproantigen ELISA method. Methods During the present investigation experimental infection was established using evaginated worms of Echinococcus granulosus (E. granulosus). To check cross reactivity two pups were infected with Taenia hydatigena (T. hydatigena). In order to detect the presence of antigen, hyperimmune sera were raised against excretory-secretory products of adult worms E. chinococcus granulosus. Faecal sample collected either from experimentally infected pups or from other sources were heated at 70 °C to detect heat stable soluble antigen. Results Pups harbouring less than 104 worms showed negative results. Samples collected from 14 days onwards from experimentally infected animals harbouring more than 104 worms showed positive value. The maximum positive samples were detected in samples collected from in and around slaughter house and the least number of samples were detected positive maintained by dog squad. Conclusions The affinity purified IgG exhibited promising results for detection of canine echinococcosis by indirect ELISA.

527-530

Authors: BS Ashok Kumar, K Lakshman, KN Jayaveera, C Vel Murgan, PA Arun Kumar, R Vinod Kumar, Hegade Meghda, SM Sridhar

Objective To investigate the analgesic activity of methanolic extract of Amaranthus viridis (A. viridis), Amaranthus caudatus (A. caudatus) and Amaranthus spinosus (A. spinosus). Methods In this study, the analgesic activity of methanol extracts of all three plants at doses of 200 and 400 mg/kg were investigated by acetic acid-induced writhings test, hot plate test and tail immersion test for mice. Results It was found that all the three plants showed significant pain management effect (P<0.01) at a dose of 400 mg/kg, but showed a less significant effect at a dose of 20 mg/kg in the entire tests used for evaluation of analgesic activities (P<0.05). Conclusions Methanol extracts of A. viridis, A. caudatus and A. spinosus show potent analgesic activities, and this study provides the scientific proof for their traditional claims.

535-538

Authors: Mohd Irfan Naik, Bashir Ahmad Fomda, Ebenezar Jaykumar, Javid Ahmad Bhat

Objective To find the effectiveness of essential oil of lemongrass for the treatment of pathogenic organisms. Methods Lemongrass oil was investigated for activity against Staphylococcus aureus (S. aureus), Bacillus cereus (B. cereus), Bacillus subtilis (B. subtilis), Escherichia coli (E. coli), Klebsiella pneumoniae (K. pneumoniae) and Pseudomonas aeruginosa (P. aeruginosa), using Agar Diffusion Method and Broth Dilution Method. Minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were determined by the Broth Dilution Method. The antibiotic susceptibility test against the test organisms was performed by Disc Diffusion Method. Results Lemongrass was found effective against all the test organisms except P. aeruginosa. Gram positive organisms were found more sensitive to lemon grass oil as compared to gram negative organisms. The test organisms were found inhibited by Lemon grass oil at lower concentrations in Broth Dilution Method as compared to Agar Diffusion Method. Conclusions The tested organisms, particularly gram-negative organisms had shown high resistance towards different antibiotics whereas they were found to be inhibited by lemongrass oil even at lower concentration. Thus lemongrass oil is effective against drug resistant organisms. It can be suggested that use of lemongrass oil would be helpful in the treatment of infections caused by multidrug resistant organisms.

544-546

Authors: B Valentin Bhimba, J Meenupriya, Elsa Lycias Joel, D Edaya Naveena, Suman Kumar, M Thangaraj

Objective To explore antibacterial activity and characterization of secondary metabolites isolated from mangrove plant Avicennia officinalis (A. officinalis). Methods In the present study the leaf extracts of A. officinalis were examined for its antibacterial potential using five different solvents against some reference strains of human pathogenic bacteria for the crude extract. Maximum activity was observed for ethyl acetate and hence different concentrations like 15 μL, 25 μL, and 50 μL of ethyl extracts was checked for its antibacterial activity. Partial purification of crude extract was carried by column chromatography and fractions were analyzed using gas chromatography-mass spectrometry (GC-MS) to identify compounds. Results The crude ethyl acetate extracts of A. officinalis showed remarkable antibacterial activity with zones of inhibition of 13 mm against Eschericia coli (E. coli) and 11 mm against Staphylococcus aureus (S. aureus). Fraction 13 (ethyl acetate÷methanol= 8÷2) as the most potent one against with the minimal inhibitory concentration of 30 mm against E. coli and 25 mm against S. aureus. The GCMS resultsof active column fraction (F13) revealed that the active principals were a mixture of hydroxy-4 methoxybenzoic acid, diethyl phthalate, oleic acid. Conclusions The leaf extracts with proven antibacterial effects can clearly be directed towards cancer treatment as to inhibiting cancer cell growth. The limited number of test organisms owes to a constraint of resource. So, the effect of strong bursts of leaf extracts on human pathogenic bacteria should further be tested on a wide range of test organisms.

552-554

Authors: A Mariana, SK Heah, AL Wong, TM Ho

Objective To determine distribution of arthropods in processed rice products such as rice flour and rice cereal-based infant food. Methods Random samples of rice flour and rice cereal-based infant food purchased from commercial outlets were examined for the presence of arthropods using a modified Berlese Tullgren Funnel Method. Mites were mounted prior to identification and weevils were directly identified. Results For non-expired products, infestation was found in 6.7% of rice flour and none was found in rice cereal-based infant food samples. The arthropods found in the flour samples were Cheyletus spp., Suidasia pontifica (S. pontifica), Tarsonemus spp., Tyrophagus putrescentiae (T. putrescentiae), Sitophilus granarius (S. granarius) and Sitophilus oryzae (S. oryzae). Others which cannot be identified were Oribatid and Prostigmatid mites. The most common mites in rice flour were Tarsonemus spp. (69.1%), followed by S. pontifica (18.2%). For expired products, only one sample of rice cereal-based infant food was infested and the infestation was by mites of the family Tydeidae. Conclusions This study demonstrates the presence of 4 allergenic species of S. pontifica, T. putrescentiae, S. granarius and S. oryzae in rice flour. These arthropods can contribute to the incidence of anaphylaxis upon consumption by atopic individuals. There was no infestation of arthropods in rice cereal-based infant food surveyed except for an expired product in a moderate rusty tin container.

560-562

Authors: Peter George, Lobo Manuel Alexander

Objective To identify cases of severe Plasmodium vivax (P. vivax) mono-infections among adults. Methods In this retrospective study, 30 adult patients admitted to medical wards of a tertiary hospital in a malaria endemic urban area from March 2010 to April 2010 were included. The diagnosis of P. vivax malaria was established by peripheral blood film (PBF) examination, and severe malaria was categorized as per World Health Organization guidelines. Results Complications observed were thrombocytopenia in 28 (93.3%), hepatic dysfunction and jaundice in 13 (43.3%), renal dysfunction in 8 (26.7%), severe anaemia in 3 (10.0%), cerebral malaria in 2 (6.7%), and acute respiratory distress syndrome (ARDS) in 1 (3.3%) of 30 patients. Conclusions P. vivax malaria with severe complications is common in the investigated area, and an intensive and large-scale study of the disease is necessary.

567-570

Authors: EI Ekanem, TU Agan, EE Efiok, MI Ekott, E Okodi

Objective To assess the prevalence of anemia and asymptomatia malaria parasitemia and the effect of prior antimalarials therapy on the parasite density in pregnant women at their first antenatal visit at the secondary level health care facility in Nigeria. Methods This cross sectional observational study was carried out in the antenatal clinic of General Hospital, Ikot Ekpene, Akwa Ibom State, Nigeria for 3 months period (1st June to 31st August, 2009). Five hundred and fourteen women attending their first antenatal registration visits in the hospital were recruited in the study. Socio-demographic information was obtained using pre-tested questionnaires. The malaria parasite was obtained by examining thick and thin blood films prepared on 2 glass slides while the hematocrit was obtained through 2 capillary tubes read by a Hawksleys microhematocrit reader. Results A total of 514 pregnant women participated in the study with a mean maternal age of 21.4 years and a mean gestational age at booking of 18.3 weeks. The primigravid women booked at significantly lower gestational age than multigravidae (16.2 weeks vs 21.6 weeks). Most of the women (59.3%) were anemic, out of which 60.4% were primigravida. More than half of the women had moderate to high parasite density and only 6.8% had no malaria parasitemia. All patients with severe anemia were parasitemic. Out of the 479 (93.2%) women with parasitemia, a third had taken antimalarial drugs. A majority (60.3%) of those without prior antimalarial drugs had moderate to high density parasitemia. Conclusions This study shows high prevalence of anemia in women with asymptomatic malarial parasitemia, particularly the primigravida. The severity of anemia is directly related to the density of malaria parasitemia. Those with effective antimalarial therapy appear to have low density parasitemia and therefore mild anemia. Routine screening for anemia and malaria parasites at booking, prompt parasite clearance and correction of anemia would reduce the associated maternal and perinatal complications.

574-576

Authors: Fatima Shujatullah, Haris M Khan, Razia Khatoon, Tamkin Rabbani, Abida Malik

Objective To determine the prevalence of bacterial vaginosis (BV) in patients with vaginal discharge and evaluate the efficacy of OSOM BV blue test in diagnosis. Methods OSOM BV blue test, a rapid diagnostic test based on the principle of detection of bacterial sialidase activity in vaginal fluid specimens was conducted. A total of 405 patients in the reproductive age group (15–45 years) having vaginal discharge were included in the study along with 10 healthy age-matched controls. Two high vaginal swabs were collected aseptically from each patient. One swab was used to make smear for gram staining, and the other was for OSOM BV blue test. Amine test and vaginal pH test were taken as well. Results The prevalence of bacterial vaginosis was 60.7%. OSOM BV blue test showed good efficacy, as compared with gram staining in diagnosing BV. The sensitivity and specificity of OSOM BV blue test were 97.6% and 97.5% respectively. Amsel's criteria diagnosed 180 (44.4%) cases of BV and had sensitivity and specificity of 67.1% and 90.6% respectively. Thus the performance of OSOM BV blue was better than the methods based on Amsel's criteria. Conclusions OSOM BV blue test is an efficacious bed side test, helpful in rapidly making an accurate diagnosis of BV in setups lacking laboratory facilities or expert microbiologists.

580-583

Authors: JO Odigie, KC Anugweje

Objective To determine the prevalence of asymptomatic bacteriuria in pregnancy, to consider the antimicrobial sensitivity patterns of involved uropathogens, to elucidate the safety profiles of antibacterial agents, and to evaluate the role of urinalysis in screening for asymptomatic bacteriuria. Methods About 760 apparently healthy pregnant subjects attending the Antenatal Clinic of the University of Port Harcourt Teaching Hospital were randomly selected for this study. Urinalysis and microscopy, culture, and sensitivity tests were carried out on clean-catch midstream urine samples obtained from subjects. Biochemical reagent strips were used for urinalysis while the standard wire loop and agar diffusion technique were respectively employed for culture and susceptibility testing. Results A total of 111 samples yielded moderate or severe growth on culture after 48 hours comprising 35, 31, 27, and 18 isolates of Staphylococcus spp., Proteus spp., Klebsiella spp., and Escherichia spp, respectivehy. Urinalysis results were positive for the presence of nitrate reductase and leucocyte esterase activity in 17 urine samples of these 111 samples. The isolates showed a general sensitivity to the fluorinated quinolones and to Nitrofurantoin. Conclusions The prevalence of asymptomatic bacteriuria is 14.6%, with the predominant organism being Staphylococcus spp. Drugs used for treatment should have excellent fetal safety profiles, and a rapid screening test with a high negative predictive value for asymptomatic bacteriuria would be ideal.

586-588

Authors: BDSS Budagoda, KAS Kodikara, WKS Kularatne, RM Mudiyanse, DH Edussuriya, JP Edirisinghe, IP Karunaratne, KGAD Weerakoon, SC Medagedara, SAM Kularatne

The sting of Giant Asian honeybee (Apis dorsata) or Bambara in Sinhala and Karunge Kulavi in Tamil is a common environmental hazard in Sri Lanka known to cause immediate allergic reactions, which could be fatal in sensitized individuals. We reported myocardial infarction, bowel gangrene and fatal anaphylaxis in a prospectively proven case series and the association of these uncommon complications with delayed removal of stingers from the patients' skin.

427-434

Authors: A Mariana, MN Shukor, Norhazizi H Muhd, Nurlemsha B Intan, TM Ho

Objective To document movement patterns, home range, nesting behaviour and social organization of 5 individuals (3 males and 2 females) of a common species of tree-shrew, Tupaia glis (T. glis) surrounding houses of otoacariasis cases. Methods Each shrew was fitted with a transmitter chip radio-collar which operates between the frequencies of 154.13 MHz to 154.21 MHz. Each transmitter was then tracked with a Portable Telemetry Receiver (Sirtrack, New Zealand) fitted with a 3-element Yagi antenna. Collared shrews were located using standard methods of ground-based triangulation. Each location was taken from at least 2 directional fixes and a minimum of 3 compass bearings. Fixes were taken hourly for each collared individual from the time of emergence from nest (beginning of activity) till time of entry into the nest (end of activity) every day for 5 to 7 continuous days. Three series of radio telemetry observations were carried out. The bearings, time and positions of an observer were recorded and later plotted on a graph paper in order to derive coordinates of the collared animal. [These coordinates then analyzed using Ecological Software Solutions (Biotas Version 1.03)]. Results Nests were found in a jack fruit tree, long bushes, and 2 houses. Daily telemetry detections demonstrated 2 individuals of different sex having nests (or a nest) in the same house. All shrews emerged from and returned to their nests between 0601 to 0659 hours and 1901 to 1959 hours, respectively. Both the time of exit from and entry into nest were the same between sexes (P>0.05). Their average total active period was 4.90 to 7.00 hours with a total daily travel distant of 270 m to 382 m. A male and a female shrew can move as far as 3 285 m and 4 591 m, respectively. Active movements of T. glis were during daytime. They regularly entered some houses in the area during day and night except for one individual which visited during daytime only. The sizes of home range and core area for the shrews were 2.00–3.40 ha and 0.05–0.42 ha, respectively. Generally, the mean home range size of females was 20.8% larger than that of males. Females covered a 15.4% slightly higher daily movement range compared to males. Conclusions This is the first radio telemetry study in Malaysia to monitor movements and home range of shrews carrying ticks on their body. It demonstrates that shrews are potential carriers of ticks from wild into the houses and their compounds based on their total active periods spent moving around from fruit orchards, secondary forest, plantations and other vegetations to trees in compound of 4 to 7 houses and vice versa. There are also evidences showing shrews have close contact with humans.

439-442

Authors: Amin Mansour, Kalantar Enayat, Mohammad-Saeid Neda, Ahsan Behzad

Objective To evaluate the antibacterial effect and physicochemical properties of essential oil of Zataria multiflora Boiss. Methods Antibacterial activity of essential oil of Zataria multiflora Boiss was assessed by agar disc diffusion and minimal inhibitory concentration(MIC) was tested by E test. Results The essential oil of Zataria multiflora Boiss was effective on pathogenic bacteria particularly Staphylococcus aureus (S. aureus). The MIC values of the target cultures ranged from 0.39 mg/mL to 1.56 mg/mL. The physicochemical properties like effects of pH, temperature, detergents, and enzymes on the activity of essential oil from Zataria multiflora Boiss were also determined. The essential oil was quite stable to temperature as tested against S. aureus and Escherichia coli (E. coli). The essential oil was very stable over a wide range of pH. The antibacterial activity of essential oil was insensitive to various protein-denaturing detergents (Such as Tween 80, Tween 20, Triton 100, etc.) and enzymes (namely proteinase K, trypsin, lipase, and lysosyme). Conclusions A potential use of the essential oil from Zataria multiflora Boiss is suggested. More studies including further purification, mass spectra, nuclear magnetic resonance (NMR) and evaluation of toxicity are needed for confirmation of this suggestion.

448-450

Authors: Onyebiguwa Patrick Goddey Nmorsi, Clement Isaac, Igho Benjamin Igbinosa, Duncan Ogheneocovo Umukoro, Dafe Palmer Aitaikuru

Objective To investigated the prevalence of human African trypanosomiasis (HAT), a neglected tropical disease caused by Trypanosoma brucei gambiens in an endemic focus of Nigeria, as it relates to age, sex and occupational differences. Methods A total of 474 human subjects were screened using card agglutination test for trypanosomiasis kit. Positive samples were further investigated for parasite positivity in blood/serum and cerebrospinal fluid (CSF). Results Of the 474 screened, 44(9.3%) were seropositive with seroprevalence of 22(9.6%) in Urhouka, 14(9.5%) in Umeghe and 8(7.9%) for Ugonu. The number of seropositives, observed for weakly, moderately and strongly positives for the three communities were 4, 7 and 11 in Urhouka, 4, 5 and 5 in Umeghe and 3, 2 and 3 in Ugonu respectively. Among the 16 volunteers with detected parasite in their blood, 4 of them were weakly positive, 5 of them were moderately positive and 7 of them strongly positive. 4 volunteers from Urhouka community were found parasites in their CSF and they were all strongly positive. The difference between the seroprevalence of males and females was not statistically significant (OR=1.14, 95% CI=0.37–3.4, P>0.05). The prevalence difference between age group 21–30 years old and the youngest and oldest age groups was statistically significant (OR=3.5, 95% CI=1.08–12.57, P<0.05) but not significant for other age categories (P>0.05). It was observed that farmers had significantly higher prevalence of HAT infection as well as greater risk of Trypanosoma brucei gambiense infection than inhabitants with other occupations (OR=3.25, 95% CI=0.99–11.79, P<0.05). Conclusions Human activities such as farming and visits to the river have been identified as major risk factors to HAT. Also the breakdown of HAT control program has been advanced for the rise in HAT in Abraka, an endemic focus in Nigeria.

454-457

Authors: PN Krishnan, R Saraswathi, C Dilip, N Ramarao

Objective To formulate and evaluate acyclovir microcapsules using bakers yeast. Methods Acyclovir, pretreated yeast and deionised water were taken at a volumetric ratio of 1:2:4 respectively. This suspension was agitated in a magnetic stirrer at 25°C, 30°C, 35°C, and 40°C for 4 hours. The suspension was then centrifuged for 10 minutes at 2 000 rpm. The supernatant solution was decanted and the cells were washed 5 times with deionised water. Then the suspended drug entrapped yeast cells were dried in a lyophillizer for 48 hours. The yield was noted. Results The first four formulations were done with 200 mg of the drug, followed by 400 mg for the next four formulations and 800 mg the last four formulations. SEM showed that the surface of the microcapsules was intact, with no burst characteristics. FTIR showed no interaction between acyclovir and the cell wall. DSC showed that the peak was within the standard values. The mean particle size for all the samples was 8 μm in diameter. The dissolution studies were done for all the twelve samples and showed a Fickian model of diffusion. Conclusions From the results it is inferred that the samples prepared at 40°C (FY-4, FY-8, FY-12) show better entrapment and release. So these samples are formulated in the form of a suspension and compared with marketed acyclovir suspension using HPLC technique. The formulated suspensions with FY-4, FY-8 and FY-12 shows drug content in accordance with the standards of the pharmacopoeial limits.

461-464

Authors: Momohmumuni Audu, Adikwu Michael Umale, Ibezim Emmanuel Chinnedu

Objective To formulate and evaluate the application of PEGylated mucin in oral bioadhesive drug (OBD) delivery system. Methods The bioadhesive strength of different formulation ratios of polyethylene glycol (PEG) and mucin was evaluated by tensiometry. Co polymer matrices containing mucin and PEG (PEGylated mucin) in the following ratios of 1:1 (A), 2:1 (B), 1:2 (C), 0:1 (D) and 1:0 (E) were prepared by co-precipitation. Microparticles were prepared from the matrices using a size reduction technique. Five different samples were prepared using direct mixtures in the appropriate solvenfs having mucin and PEG ratios of 1:1, 2:1, 1:2, 0:1, 1:0 and labeled as (A-E). The matrices were evaluated for their flow properties and the in vitro bioadhesion characteristics of the samples were examined. Results The range of flow rates of the matrices was 1.35–2.23 g/sec. The angle of repose was in the range of 37.3–39.9 degree. The bulk and tapped densities were within the ranges of 0.41–0.49. The Hausner's quotient (HQ) were 1.12, 1.24, 1.14, 1.25 and 1.2 for matrix batches A-E respectively, The bioadhesive strength of polymer matrices appeared to be directly related to amount of mucin. The order of bioadhesive strength is 2:1 > 1:1 > 1:0 > 1:2 > 0:1 of PEG: mucin in both simulated intestine fluid (SIF) and simulated gastrointestinal fluid (SGF). The physical properties of the micrometrics properties of the matrices were within the accepted values. Conclusions OBD preparation containing PEGylated mucin can be prepared by direct compression and be used in drug delivery to the oral cavity.

471-474

Authors: Abhishek K Jain, V Ravichandran, Madhvi Sisodiya, RK Agrawal

Objective To synthesis 2–substituted–4,5–diphenyl–N–alkyl imidazole derivatives. and evaluate their antibacterial activity. Methods A mixture of benzil (10 mmol) and ammonium acetate (0.1 mol) (immediately fused) in glacial acetic acid (25 mL) was stirred at 80–100 °C for 1 h under nitrogen atmosphere (to prevent incorporation of any atmospheric impurities and moisture). Substituted aldehydes (10 mmol) in glacial acetic acid (5 mL) was added drop-wise over a period of 15–20 min at the same temperature and stirred for another 4 h, the progress of the reaction was monitored by TLC test using ethyl acetate as eluent. The newly synthesized compounds were characterized by IR, 1HNMR, 13CNMR and by mass spectroscopy. Results All the synthesized compounds were confirmed by spectroscopical techniques and evaluated for antimicrobial activity against Staphylococcus aureus(S. aurius), Bacilus subtilus (B. subtilus), and Escheria coli (E. coli). These compounds showed antibacterial activity (zone of inhibition) against S. aurius ranged from 3 mm to 9 mmin diameter, B. subtilus, 4–8 mm, and E. coli 5–12 mm. Out of 2a-2e, only 2a and 2b showed some sort of activity but none of them had considerable activity compared with that of the standard. Conclusions All the synthesized compounds show moderate activity against the tested bacteria S. aurius, B. subtilus, and E. coli. So, further structural modification is necessary to improve the antibacterial action of 2-substituted-4,5-diphenyl-N-alkyl imidazole derivatives.

479-482

Authors: GT Jombo, S Akpan, J Epoke, Akaa P Denen, F Odey

Objective To ascertain the antimicrobial susceptibility profile of Pseudomonas aeruginosa (P. aeruginosa) recovered from surgical site infections (SSIs). Methods The study was retrospective in nature and was compiled for a period of five years (1st February, 2004–31st January, 2009). Data were generated from the culture of post-operative wound swab specimens by the microbiology laboratory of University of Calabar Teaching Hospital. Relevant information from the patients' records was compiled, such as age, gender, type of surgical procedure, microorganisms recovered and their antibiotic sensitivity patterns. Obtained data was analysed by using Epi Info 6 statistical software. Results Of the 4 533 wound swab specimens processed, 673 were culture positive and P. aeruginosa was recovered from 13.1% of the culture positive specimens with its rate of recovery decreasing with age progression (P< 0.05) but with no gender difference (P> 0.05). Most of the P. aeruginosa isolates were from general surgery wards and least from orthopaedic wards. Ofloxacin, ceftriaxone and augmentin were the most active antibiotics while ampicillin, tetracycline and cotrimoxazole were the least active antibiotics, with no antibiotic having a 100% activity against the organism. Conclusions In view of the high resistance displayed by P. aeruginosa recovered from SSIs, adequate antiseptic procedures should be entrenched to avoid colonization of surgical wounds by this microorganism as well as others with similar sensitivity profile. Ofloxacin, ceftriaxone and augmentin may be considered for prevention of P.aeruginosa infection.

486-490

Authors: Neeraj Tiwari, Vineeta Kandpal, Ajoy Tewari, K. Ram Mohan Rao, VS Tolia

Objective To investigate the presence of statistically significant geographical clusters of tuberculosis (TB) using Geographical Information System and spatial scan statistics in Dehradun, India. Methods The spatial scan statistic implemented with a software program, SaTScan v6.1, was used to test the presence of statistically significant spatial clusters of TB and to identify their approximate locations (P< 0.05 for primary clusters and P<0.1 for secondary clusters). Geographical Information System was used for geographical analysis. Results Significant high rate spatial clusters were identified in seven wards of the Dehradun Municipal area. Conclusions There is sufficient evidence about the existence of statistically significant TB clusters in seven wards of Dehradun, India. The purely spatial scan statistics methodology used in this study has a potential use in surveillance of TB for detecting the true clusters of the disease.

494-495

Authors: Krishnananda Prabhu, Gummadi Maheshwar Reddy, Anjali Rao

Objective To measure and compare pre- and post-treatment levels of serum total glutathione-S-transferase (GST) in stage IV cervical cancer patients and erythrocytic glutathione (GSH) and malondialdehyde (MDA) and plasma ceruloplasmin (CP) and total GST in stage IV oral cancer patients and to correlate with the response to treatment during a two year follow-up period in respective cancer groups. Methods Thirty-four biopsy-proven stage IV oral cancer and cervical cancer patients (n=17 in each group) who underwent same mode of treatment were chosen for this study. Erythrocytic MDA and GSH, CP and serum total GST were measured in all patients before the onset of treatment, and the GST level was only measured in cervical cancer patients after radiotherapy. The levels were compared with their respective prognosis in 2 years. Results Oral cancer patients with higher pretreatment levels of GSH, CP and GST came with cancer recurrence within 2 years after the onset of treatment. Cervical cancer patients with higher post radiotherapy levels of serum total GST had higher recurrence tate. Conclusions This study indicates there may be a role for these antioxidant parameters namely GST, GSH, CP in assessment of long term survival and prognosis of cancer patients.

499-504

Authors: Charuai Suwanbamrung

In order to understand the community capacity for sustainable community-based dengue prevention and control, this paper proposes the approach of a previous study about meaning and domains of dengue prevention and control, an assessment tool and a community capacity building model for sustainable community-based dengue prevention and control in the Southern Thailand. A study of dengue community capacity domains was conducted by utilizing a qualitative method, whereby ten initial community domains were identified by means of a literature review, in-depth interviews of sixty community leaders, and eight focus group discussions with sixty non-leaders in four sub-districts of southern Thailand. In the final study, there were 14 identifiable domains in leaders group and 11 domains in non-leaders. The resulting dengue community capacity-assessment tool (DCCAT) consisted of two parts: one for leaders (DCCAT-L) and the other for non-leaders (DCCAT-NL). DCCAT-L was composed of 115 items within 14 domains and 83 items within 11 domains for the DCCAT-NL. The key domains of leaders and non-leaders had a partial overlap of domains such as critical situation management, personal leadership, health care provider capacity, needs assessment, senses of community, leader group networking, communication of dengue information, community leadership, religious capacity, leader group and community networking, resource mobilization, dengue working group, community participation, and continuing activities. The application of the new tool consisted of five steps: 1) community preparation, 2) assessment, 3) a community hearing meeting, 4) interventions, and 5) conclusion and improvement step. All stakeholders in the community should use the new tool based on a clear understanding of the measurement objectives, the desired outcomes, resources available and characteristics of their community. If communities need to develop and build dengue community capacity, then the designed pre-post intervention assessments or serial assessments are essential.