Asian Pacific Journal of Tropical Medicine

841-845

Authors: Wang Yalin, Zeng Lingbing, Yang Hongliang, Xu Jianmin, Zhang Xiangyan, Guo Xiaokui, Pal Utpal, Qin Jinhong

Objective To assess the prevalence of Leptospira detected in wildlife and domesticated animals in Jiangxi Province, China, in. Methods Urine samples from 28 buffaloes and kidney samples from 50 pigs, 50 dogs and 38 rats were collected from Fuliang and Shangrao County, Jiangxi Province, China, in October 2009. Polymerase chain reaction(PCR)and culture analyses were used to detect Leptospira. The cultured isolates were typed using the microscopic agglutination test(MAT). Results The results showed that rats potentially serve as the main reservoir of leptospiral infection, followed by dogs. Although 16% of rats (6/38) were positive using culture analysis, PCR analysis using the diagnostic primers G1/G2 and B64I/B64II or lipL32 showed identification as 50% and 24%, respectively, of the rat samples as positive for the presence of leptospiral DNA. Conclusions PCR-based detection of leptospiral DNA in infected kidney tissues of reservoirs is more efficient when using G1/G2 primers than lipL32 primers. However, the latter primers have a potential application for detection in urine samples. The alarmingly high prevalence of leptospiral DNA in the wild rat population near human habitation underscores the utility of routine Leptospira surveillance, preferably using PCR methods, which are more sensitive than traditional culture-based methods.

850-856

Authors: Chen Jinling, Zhu Dandan, Shen Pei, Sun Wei, Xiao Gengfu, Duan Yinong, Zhu Ying

Objective To analyze the sequence of ORF1 protein of Torque teno virus to prepare for the future hybrid experiments. Metheds The sequence of ORF1 protein of Torque teno virus was analyzed by bioinformatics using some web tools. Results The most likely cleavage site was between position 14aa and 15aa and signal peptide may be position 1aa–14aa. Two possible transmembrane helices from inside to outside and three possible transmembrane helices from outside to inside were found. The position 509 (NKTN) was the potential N–glycosylation site. The speculative molecular weight of TTV ORF1 protein, which may be a kind of unstable protein was 88 705.7 Da. 1aa–91aa and 278aa–361aa were localized in non–regular secondary structure region. Conclusions TTV ORF1 protein may be a nuclear protein which contains two non–regular secondary structure region. 265aa to 486aa and 510aa to 679aa may be the two approciate fragments to construct the plasmids, which would be prepared for the future hybrid experiments to study the functional positions of the protein and the interactions between TTV and its hosts. Bioinformatics analysis would possibly make it easier to study the protein's function.

862-865

Authors: S Ravikumar, G Ramanathan, M Gnanadesigan, A Ramu, V Vijayakumar

Objective To identify the in vitro antiplasmodial activity of seaweed plants against Plasmodium falciparumstrains. Methods A total of eight seaweeds were collected from Kanyakumari district, Tamilnadu, India. The in vitro antiplasmodial activity was performed in 96 well plates against Plasmodium falciparum, and preliminary phytochemcial analysis were performed for the extracts. Results Of the selected plants Enteromorpha intestinalis (2.61%) showed maximum percentage of extraction. The minimum concentration of inhibitory (IC50) value was observed with Chaetomorpha antennina [(26.37±4.14) μg/mL] further, the positive controls such as chloroquine and artemether showed antiplasmodial activities (IC50) with (19.10±5.93) and (6.03±0.21) μg/mL concentrations, respectively. The preliminary phytochemical analysis of the seaweed extracts showed a variety of phytochemical constituents such as carboxylic acids, phenols, protein, resins, steroids and sugars. Conclusions The antiplasmodial activity of the seaweed extract might due to the presence of sugars and phenolic compounds. From the present findings, it is concluded that, the seaweed extract of Chaetomorpha antennina can be further used as a putative antiplasmodial drugs in near future.

870-877

Authors: Puthuparampil Nazarudeen Ansil, Anand Nitha, Santhibhavan Prabhakaran Prabha, Pallara Janardhanan Wills, Vahab Jazaira, Mukalel Sankunni Latha

Objective To identify the phytochemical constituents of Amorphophallus campanulatus (A. campanulatus) tuber and to evaluate its antioxidant potential through in vitro and in vivo models. Methods Phytochemical screening and in vitro antioxidant activities of A. campanulatus tuber n–hexane extract (ACHE) and methanolic extract (ACME) were evaluated using DPPH, hydroxyl radical, reducing power and total antioxidant capacity assays. The total phenolic and flavonoid contents were also investigated. The protective potential of two different doses of ACME (125 and 250 mg/kg) was also evaluated against thioacetamide (TAA) induced oxidative stress in rats. Silymarin used as a standard drug control. Hepatotoxicity was assessed by quantifying the serum levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP) and lactate dehydrogenase (LDH). The antioxidant potential of ACME were also evaluated by the estimation of catalase (CAT), glutathione peroxidase (GPx), glutathione reductase (GR), glutathione-S-transferase (GST), reduced glutathione (GSH) and lipid peroxidation (Thiobarbituric acid reactive substances) in hepatic and renal tissues. Histopathologic changes of liver were also evaluated. Results In vitro studies revealed that ACME has higher antioxidant and radical scavenging activity than ACHE, which may be attributed to its higher phenolic and flavonoid content. ACME significantly prevented the elevation of serum AST, ALT, ALP, LDH, and tissue malondialdehyde levels(P < 0.05). Hepatic and renal GSH, GST, GR, GPx, and catalase levels were remarkably increased by the treatment with the extract. Quantification of histopathological changes also supported the dose dependent protective effects of ACME. Conclusions The results do suggest that A. campanulatus tuber could be considered as a potential source of natural antioxidant.

883-888

Authors: Rajesh J Mandade, SA Sreenivas, DM Sakarkar, Avijit Choudhury

Objective To explore the effects of extract of Hibiscus rosasinensis (H. rosasinensis) on the volume, free and total acidity of gastric secretion induced by carbachol. Methods Animals were kept on fasting for 48 h, then the pylorus of each animal was ligated. They were randomly divided into 5 groups and treated by carbachol at 600 μg/kg. Then animals in group II – V were treated by H. rosasinensis extract at 250 and 500 mg/kg body weight, cimetidine at 2.5 mg/kg and verapamil at 10 mg/kg body weight intraperitoneally, respectively. The volume, free and total acidity of gastric secretion were observed and compared. Results It was found that the extract significantly reduced the volume, free and total acidity of gastric secretion (P<0.01). These reductions were comparable to cimetidine and verapamil. And the reduction in the volume and free acidity were more significant in cimetidine and verapamil treated group indicating that cimetidine and verapamil were more effective. Conclusions The extract of H. rosasinensis can reduced the volume, free and total acidity of gastric secretion, and can be used effectively in the treatment of peptic ulcer.

900-903

Authors: Dinesh Kumar, Sunil Kumar, Sonia Kohli, Renu Arya, Jyoti Gupta

Objective To evaluate the antidiabetic potential of methanolic extract of Albizia odoratissima Benth. bark in alloxan induced diabetic mice. Methods Group-I(normal control) mice received only basal diet without any treatment. In Group– II(Diabetic control) mice, diabetes was induced by alloxan (150 mg/kg i.p.) and received only Tween 80, 5% v/v in normal saline. Group– III and Group– IV mice received metformin (10mg/kg) and gliclazide (10 mg/kg) as standard drugs. Group– V and VI mice received methanolic bark extract of Albizia odoratissimaat doses of 250 and 500 mg/kg body weight p.o., respectively. Results The results of the study indicates that Albizia odoratissima bark extract significantly (P<0.01) reduced the blood sugar level. The bark extract also significantly reduced the levels of serum cholesterol, triglycerides, serum glutamic–oxaloacetic transaminase, serum glutamic–pyruvic transaminase, alkaline phosphatase and decreases level of total proteins in alloxan induced diabetic mice. Conclusions Methanolic extract of Albizia odoratissima has protective effects on the protection of vital tissues (pancreas, kidney, liver, heart and spleen), thereby reducing the causation of diabetes in experimental animals.

910-916

Authors: Sachin L Badole, Anand A Zanwar, Abhijeet N Khopade, Subhash L Bodhankar

Objective To evaluate the in-vitro antioxidant and antimicrobial activity of cycloart–23-ene–3β, 25-diol (called as B2) isolated from stem bark of Pongamia pinnata. Methods In vitro antioxidant activity of B2 was determined by methods for determination of DPPH radical scavenging, reducing power, superoxide anion radical scavenging, hydroxyl radical scavenging, hydrogen peroxide scavenging, metal chelating and nitric oxide radical scavenging at the doses of 20, 40, 60, 80 and 100 μg/mL, respectively. β-tocopherol with same concentration was used as a standard antioxidant. In vitro antimicrobial activity of B2 was determined by cup plate method in different concentration range of 10-100 μg/mL. Results The results indicated that dose dependent % reduction against DPPH radical, reducing power, superoxide anion radical scavenging, hydroxyl radical scavenging, metal chelating, hydrogen peroxide scavenging and nitric oxide radical scavenging by B2 and β-tocopherol. Conclusions It is concluded that cycloart 23-ene-3β, 25 diol (B2) showed dose dependent antioxidant activity. B2 showed more DPPH radical scavenging, reducing power, superoxide scavenging, hydroxyl radical scavenging, metal chelating scavenging, hydrogen peroxide radical scavenging and nitric oxide radical scavenging activity than β–tocopherol and in case of antimicrobial activity B2 exhibited broad-spectrum activity against bacteria and strong activity against yeast type of fungi.

921-924

Authors: Krishna Murti, Upendra Kumar, Mayank Panchal

Objective To screen the wound healing activity of aqueous and ethanolic extract of roots of Ficus benghalensis. Methods Both the extracts were evaluated for wound healing by three modelsie. incision, excision and dead space wound. In incision and dead space the extracts were applied daily topically till the 10 th post wounding days while in excision model it was till the complete epithelialization process. Standard group were administered Povidone iodine ointment topically daily. The breaking strength, percentage of wound contraction, period of epithelialization, dry granulation weight and hydroxyproline content were observed. Results The result of the present study showed that both extracts were able to increase the breaking strength (incision model), decrease period of epithelialization, increase percentage wound contraction (excision model), increase hydroxyproline content (Dead space wound model) significantly compared with control group (P<0.05). Conclusions Based on result we conclude that aqueous extract is more effective than ethanolic extract. However, it is needed more research to be carried out especially on toxicity studies of ethanolic extract.

757-759

Authors: Viroj Wiwanitkit

Leishmaniasis is an important tropical vector-borne disease. This infection can be seen in tropical area and it is considered to be one of the most important vector-borne infections at present. The general situation of the leishmaniasis in Thailand is hereby reviewed. Although Thailand is a tropical country, the leishmaniasis is not endemic but sporadic. The imported cases are documented in some literatures. The serious form of leishmaniasis, the visceral leishmaniasis is also detectable in Thailand. Also, the author performed an in depth literature review of the reports of bone marrow leishmaniasis, a specific kind of visceral leishmaniasis, in Thailand in order to summarize the characteristics of this infection among Thai patients. According to this review, there have been at least 5 reports in the literature of 6 cases of bone marrow leishmaniasis in the Thai population, of which no case was lethal. Concerning the clinical manifestations, all except had prolonged fever with unknown origin. From physical examination, all had hepatosplenomegaly. The striking findings were active hemophagocytosis with increased proliferation of lymphoidplasma cell line in the bone marrow and amastigotes of Leishmania donovani was demonstrated. Considering the treatment, pantavalent antimony compound was used and the excellent improvement and complete recovery. Finally, the author also discussed on the importance of leishmaniasis in Thailand relating to the present globalization and good traveling system.

764-768

Authors: Anuradha Singh, Mohd Shahid, Farrukh Sobia, Umesh, Haris M Khan

Objective To determine the occurrence of class A and class C β-lactamase genes and their co-occurrence in Indian Enterobacteriaceae. Methods 52 third generation cephalosporin resistant isolates were phenotypically detected by combination disk method and screened by PCR to identify class A and class C type β-lactamase genes. Results Of the 52 isolates, 94.2% (49) were found harboring any of the blaESBLs. blaCTX-M, blaSHV and blaTEM were present in 82.6% (43/52), 59.6% (31/52), and 42.3% (22/52) isolates, respectively. Of the 49 ESBL positive isolates 57.1% (28/49) showed co-occurrence of blaampC with blaESBLs. On the contrary, the collection from 2009 showed their co-occurrence in 81.4% isolates. Conclusions The comparative study shows a downward trend for co-existence of blaESBLs with blaampC from 2009 to 2010. Further large scale studies are needed to address the co-occurrence of class A and class C β-lactamases in India and the resistance trend occurring over a period of time.

773-777

Authors: Salma Medani Elhag, Eshtiyag Abdalla Abdelkareem, Ashraf Siddig Yousif, Ehab Ahmed Frah, Ahmed Babiker Mohamed

Objective To determine secreted antischistosoma antibodies in urine and to discern the epidemiological situation of schistosomiasis in the agricultural field labourers'camps city in the Gezira State-central Sudan. Methods Total of 66 urine and 66 serum paired samples were collected from those who confirmed parasitologically positive and negative with schistosomiasis from the two camps. Samples were tested using ELISA technique to measure and compare the immunoglobulin G (IgG) levels in serum and urine samples of schistosomiasis patients. Results The overall prevalence of S. mansoni and S. haematobium was 53.8% and 15.4%, while the intensity were (2.04 GMEC) and (0.9 GMEC) respectively. The relative percentage of positive IgG individulas in urine was 92.40% where as 96.97% in serum. Statistically no significant difference between the IgG levels in serum and urine samples was observed. Conclusions This study shows that the detection of secreted IgG antibodies in urine can substitute serum for diagnosis of schistosomiasis.

786-790

Authors: M Rowshahul Habib, M Rezaul karim

Objective To investigate experimentally the possible antitumor effect of methanol extract (ME) of Calotropis gigantea L. (C. gigantean) root bark and its petroleum ether (PEF) and chloroform (CF) soluble fractions against Ehrlich ascites carcinoma (EAC) in Swiss albino mice. Methods The effects of ME (10 and 20 mg/kg), PEF (40 and 80 mg/kg) and CF (20 and 40 mg/kg) on the growth of EAC and life span of EAC bearing mice were studied. Hematological profile and biochemical parameters (SALP, SGPT and SGOT) were also estimated. Results Results of in vivo study showed a significant decrease in viable tumor cell count and a significant increase of life span in the ME and CF treated group compared to untreated one. The life span of ME and CF treated animals was significantly (P<0.05) increased by 43.90% (20 mg ME/kg) and 57.07% (40 mg CF/kg). ME and CF brought back the hematological parameter more or less normal level. ME and CF also restored the altered levels of serum alkaline phosphatase (SALP) and serum glutamate oxaloacetate transaminase (SGOT). Conclusions Methanol extract (ME) of C. gigantea root bark and its chloroform soluble fraction (CF) possesses significant antitumor activity.

796-798

Authors: Chinaka O Nwaehujor, Nkeiruka E Udeh

Objective To explore the hepatoprotective and anti-oxidant activities of the methanolic leaf extract of Bridelia micrantha (B. micrantha) on paracetamol induced liver damage in Wistar rats. Methods Parameters were measured including alanine aminotransaminase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), bilirubin and total protein. The anti-oxidant effects were studied using the 1, 1-Diphenynl-2-Picrylhydrazyl (DPPH) and Ferric Reducing Antioxidant Power (FRAP) assay methods. Results B. micrantha extract decreased the level of AST in the rats given PCM from (129.47±0.92I) IU/L to (57.78±1.71) IU/L (P<0.05). This was lower than the value for Silymarin which was (59.92±1.41) IU/L. ALT concentration was reduced from (150.18±2.23) IU/L to (79.10±2.01) IU/L (P<0.05). ALP was reduced from (49.86±0.85) IU/L to (29.64±1.53) IU/L (P<0.05). Total bilirubin was reduced from (2.14±0.10 mg/dL) to (0.18±0.07) mg/dL (P<0.05) while total protein was increased from (4.26±0.30) mg/dL to (6.20±0.19) mg/dL (P<0.05). Concentrations ranging from 10 – 400 μg/mL of B. micrantha were assayed for antioxidant activities. The DPPH assay showed 98% antioxidant activity at concentration of 400 μg/mL. The FRAP values were 0.016, 0.39, 0.455, 0.601 and 1.382 μM at 10, 50, 100, 200 and 400 μg/mL respectively. Conclusions Results suggest that B. micrantha has hepatoprotective and anti oxidant potentials. However, further work involving fractionation needs to done to isolate the active compound responsible for the hepatoprotective activity.

804-807

Authors: Parminder Nain, Ashok Kumar, Sunil Sharma, Jaspreet Nain

Objective To evaluate in vitro antimicrobial and antioxidant activities of methanolic extract of Jasminum humile (J. humile) leaves extract. Method Methanolic extract of J. humile was evaluated for its antimicrobial activity by using agar well diffusion method & their possible antioxidant assay by two complementary test systems, namely DPPH and hydrogen peroxide scavenging activity. These various antioxidant activities were compared to standard antioxidants such as ascorbic acid for both the tests. Results In the DPPH & hydrogen peroxide scavenging activity, the IC50 value of methanol extract was 70.43 μg/mL & 60.79 μg/mL respectively. Further, the extract showed inhibitory activity for Gram-positive and negative bacteria at different concentrations. The maximum antibacterial activity of extract was exhibited against Staphylococcus aureus (S. aureus) at concentration 50 mg/mL when compared with ciprofloxacin Conclusions These results clearly indicate that J. humile is effective in scavenging free radicals and has the potential to be a powerful antioxidant. Thus, the results obtained in the present study indicate that J. humile leaves extract could be considered as a potential source of natural antioxidants and that could be used as an effective source against bacterial diseases.

814-818

Authors: Dibua Esther Uju, Nnamani Petra Obioma

Objective To investigate three tropical plant materials – clove seeds [Syzygium aromaticum (S. aromaticum)], bitter kola fruits [Garcinia kola (G. kola)] and tobacco leaves (Nicotiana species) as potential targeted killers of Streptococcus mutans (S. mutans), a cavity-causing bacterium (gram-positive, facultative anaerobe) that resides in a multispecies microbial community (dental plaque) for the treatment of dental caries (tooth decay). Methods Thirty one (31) teeth samples were collected from patients with obvious signs of tooth decay (swollen gum, weak or fallen tooth, etc.) using sterile swab sticks. These samples were collected from two major dental clinics in Nsukka, Enugu State, Nigeria and investigated by spread inoculation onto sterile blood agar and Mueller Hinton agar (MHA) respectively and incubated at 37 °C for 24 h. The discrete colonies obtained were further re-inoculated onto sterile Mitis salivarius agar (MSA) plates and incubated as above. The isolates were characterized by gram staining and catalase test. Tobacco leaves, clove seeds and bitter kola fruits were ground into powder, extracted with three different solvents (n-hexane, hot water and ethanol), filtered, dried and stored in clean containers, corked and kept until used. The plant extracts were investigated for phytochemistry, minimum inhibitory concentration (MIC), minimum cidal concentration (MCC) and compared with some conventional antibiotics commonly used against tooth decay. Antibiotic sensitivity test was also carried out. The results were statistically analyzed. Results The extracts showed varied phytochemical composition but most abundantly the flavonoids. Our result also shows that females (16) have more tooth decay than males (15) and that 16 samples were very bloody while 15 were slightly bloody. The microbial characterization showed that 18 samples were catalase-positive indicating the presence of S. mutans while 13 were catalase-negative suspected to be Staphylococcus spp. The Gram reaction confirmed 13 Gram-negative and 18 Gram-positive organisms. The n-hexane extract had the best antimicrobial activity followed by the ethanol and lastly hot water. MIC showed that n-hexane clove extract had the largest inhibition zone diameter, followed by bitter kola extract and lastly tobacco extract. The antibiotic sensitivity test credited ciprofloxacin the best because it exhibited broad spectrum of action. Conclusions Since the n-hexane extract of clove seeds demonstrated preferential growth-inhibitory activity against the causal cariogenic pathogens (S. mutans) in dental caries, we therefore, report here that clove extract be henceforth considered as a potential ingredient in toothpaste preparation.

824-828

Authors: Pierangeli G Vital, Windell L Rivera

Objective To determine the antibacterial, antifungal, antiprotozoal, cytotoxic, and phytochemical properties of ethanol extracts of leaves of Voacanga globosa (Blanco) Merr. (V. globosa). Methods The extracts were tested against bacteria and fungus through disc diffusion assay; against protozoa through growth curve determination, antiprotozoal and cytotoxicity assays. Results The extract revealed antibacterial activities, inhibiting the growth of Staphylococcus aureus, Bacillus cereus, Pseudomonas aeruginosa, Micrococcus luteus, and Salmonella typhimurium. Antifungal assay showed that it inhibited Candida albicans. The antiprotozoal assay against Trichomonas vaginalis and Entamoeba histolytica showed that V. globosa can inhibit the parasites, wherein the action can be comparable to metronidazole. With the in situ cell death detection kit, Trichomonas vaginalis and Entamoeba histolytica exposed to V. globosa leaf extract was observed to fluoresce simultaneously in red and yellow signals signifying apoptotic-like changes. Preliminary phytochemical screening revealed the chemical composition of plant extract containing alkaloids, saponins, 2-deoxysugars, and hydrolysable tannins. Conclusions Thus, this study provides scientific evidence on the traditional use of V. globosa leaf extract in treating microbial diseases. Further, the leaf extract can possibly be used to produce alternative forms of antimicrobials.

836-840

Authors: Marley García, Wilmer H Perera, Ramón Scull, Lianet Monzote

Objective To evaluate the antileishmanial activity of different extracts from three Cuban Pluchea species. Methods In in vitro assays the IC50 was calculated in the promastigotes and amastigotes forms as cytotoxicity in murine macrophages. In leishmaniasis cutanea experiment, mortality, weight loss, lesion size and burden parasite were measured. Results Extracts evaluated showed inhibitive effect on growing of promastigote form; however, active extracts caused a high toxicity. Ethanol and n-hexane extracts demonstrated specific antileishmanial activity. Ethanol and n-hexane extracts from Pluchea carolinensis (P. carolinensis) caused similar inhibition against amastigote form. The intraperitoneal administration of the ethanol extract of P. carolinensis at 100 mg/kg prevented lesion development compared with control groups. Conclusions The antileishmanial experiment suggests that ethanol extracts from P. carolinensis is the most promising. Further studies are still needed to evaluate the potential of this plant as a source of new antileishmanial agents.

1-4

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

Objective To search and analyze nitric oxide synthase (NOS) and similar proteins from Plasmodium berghei(Pb). Methods The structure and function of nitric oxide synthase and similar proteins from Plasmodium berghei were analyzed and predicted by bioinformatics. Results PbNOS were not available, but nicotinamide adenine dinucleotide 2′–phosphate reduced tetrasodium (NADPH)–cytochrome p450 reductase(CPR) were gained. PbCPR was in the nucleus of Plasmodium berghei, while 134aa–229aa domain was localize in nucleolar organizer. The amino acids sequence of PbCPR had the closest genetic relationship with Plasmodium vivax showing a 73% homology. The tertiary structure of PbCPR displayed the forcep–shape with wings, but no wings existed in the tertiary structure of its' host, Mus musculus(Mm). 137aa–200aa, 201aa–218aa, 220aa–230aa, 232aa–248, 269aa–323aa, 478aa–501aa and 592aa–606aa domains of PbCPR showed no homology with MmCPRs', and all domains were exposed on the surface of the protein. Conclusions NOS can't be found in Plasmodium berghei and other Plasmodium species. PbCPR may be a possible resistance site of antimalarial drug, and the targets of antimalarial drug and vaccine. It may be also one of the mechanisms of immune evasion. This study on Plasmodium berghei may be more suitable to Plasmodium vivax. And 137aa–200aa, 201aa–218aa, 220aa–230aa, 232aa–248, 269aa–323aa, 478aa–501aa and 592aa–606aa domains of PbCPR are more ideal targets of antimalarial drug and vaccine.

13-15

Authors: Okonta Jegbefume Mathew, Adibe Maxwell Ogochukwu, Ubaka Chukwuemeka Michael

Objective To investigated the antisecrectory activities of the methanol extract, chloroform fraction and methanol fraction of Picralima nitida seeds. Methods The methanol extract of Picralima nitida seeds was fractionated into chloroform fraction and methanol fraction. They were evaluated for antiulcer activity and gastric emptying time in rats using aspirin-pylorus-ligation model. Results Oral administration of the methanol extract, chloroform fraction and methanol fraction at 1 000 mg/kg reduced gastric ulcer by 56.4%, 40.0% and 56.3%, respectively; and the fractions of the extract significantly (P<0.05) reduced gastric emptying time when compared to the control. Gastric acidity was significantly decreased when compared with saline group, 40.25 mEq/L in methanol extract, 50.0 mEq/L in chloroform fraction 51.25 mEq/L in methanol fration but had no significant effect on the gastric secretion volume. Conclusions These findings showed that methanol extract, chloroform fraction and methanol fraction of the seeds of Picralima possessed potent antiulcer properties and some antisecretory properties.

20-23

Authors: Raju N Patil, Ravindra Y Patil, Bharati Ahirwar, Dheeraj Ahirwar

Objective To evaluate antidiabetic activity of chloroform extracts of Acacia arabica bark, Benincasa hispida fruit, Tinispora cordifolia stem, Ocimum sanctum areal parts and Jatropha curcus leaves. Methods The chloroform extracts of Acacia arabica bark, Benincasa hispida fruit, Tinospora cordifolia stem, aerial part of Ocimum sanctum and Jatropha curcus leaves were evaluated at different doses (250 and 500 mg/kg body weight.) for antidiabetic potentials in alloxan induced diabetic albino rats. The extracts were administered for two weeks in different groups whereas tolbutamide (80 mg/kg body weight) was used as reference standard throughout study. Results The result of present study showed test compounds significantly decreases elevated level of serum glucose and also caused to reverse the cholesterol, triglyceride, HDL and LDL values when compared to untreated diabetic rats. Conclusions Our finding indicates that different test extracts were able to ameliorate the derangements in lipid metabolism caused by diabetes mellitus in alloxan induced diabetic rats towards normal level.

29-34

Authors: A Bagavan, A Abdul Rahuman

Objective To evaluate the mosquito larvicidal activity of plant extracts. Methods The hexane, chloroform, ethyl acetate, acetone, and methanol leaf, flower and seed extracts of Abrus precatorius (A. precatorius), Croton bonplandianum (C. bonplandianum), Cynodon dactylon (C. dactylon), Musa paradisiaca (M. paradisiaca) and Syzygium aromaticum (S. aromaticum) were tested against fourth instar larvae of Anopheles vagus (An. vagus), Armigeres subalbatus (Ar. subalbatus) and Culex vishnui (Cx. vishnui). Results The highest larval mortality was found in seed ethyl acetate extracts of A. precatorius and leaf extracts of C. bonplandianum, flower chloroform and methanol extracts of M. paradisiaca, and flower bud hexane extract of S. aromaticum against An. vagus with LC50 values of 19.31, 39.96, 35.18, 79.90 and 85.90 μg/mL; leaf ethyl acetate and methanol extracts of C. dactylon, flower methanol extract of M. paradisiaca, flower bud methanol extract of S. aromaticum against Ar. subalbatus with LC50 values of 21.67, 32.62, 48.90 and 78.28 μg/mL, and seed methanol of A. precatorius, flower methanol extract of M. paradisiaca, flower bud hexane extract of S. aromaticum against Cx. vishnui with LC50 values of 136.84, 103.36 and 149.56 μg/mL, respectively. Conclusions These results suggest that the effective plant crude extracts have the potential to be used as an ideal ecofriendly approach for the control of disease vectors. This study provides the first report on the larvicidal activity of crude solvent extracts of different mosquitoes.

41-45

Authors: Boonyanit Thaweboon, Sroisiri Thaweboon

Objective To investigate the effect of Phyllanthus emblica (P. emblica) Linn. ethanolic extract on the adhesion of Candida albicans (C. albicans) to human buccal epithelial cells (BECs) and denture acrylic surfaces. Methods Human BECs and transparent acrylic strips were pretreated with ethanolic extract solution of P. emblica fruits at concentration ranged from 18.7 to 300 mg/mL. After washing BECs and the strips were inoculated with three strains of C. albicans (ATCC 10281 and two clinical isolates) (107 cells/mL). Normal saline solution (NSS) and 0.2% chlorhexidine gluconate were used as negative and positive controls, respectively. BECs were harvested on 12 μm-polycarbonate filters (Millipore, USA). The membrane filters and the strips were stained with Gram stain. Adherent yeast cells on 100 randomly selected epithelial cells and 20 randomly selected fields on each strip were counted under microscope. The statistical significance was calculated by Kruskal-Wallis and Tukey tests at a significant level of P< 0.05. Results Significant lower numbers of all strains of yeasts adhering to BECs and acrylic strips were observed after exposure to 75–300 mg/mL of plant extract compared with NSS. Conclusions The present study demonstrates that P. emblica ethanolic extract interferes with the adhesion of C. albicans to BECs and denture acrylic surfaces in vitro.

50-53

Authors: Hemantkumar Somabhai Chaudhari, Prati Pal Singh

Objective To produce comparative data on a group of Trichomonas vaginalis clonal strains with varied drug responses using identical methods and materials. Methods Five clonal strains of Trichomonas vaginalis were isolated from reference strain using agar plate technique. The variability of growth kinetic and susceptibility of clonal strain to metronidazole, tinidazole, satranidazole and nitazoxanide were observed in 96 well microtitre plate. Results Among these clonal strains there was a good correlation between rates of growth with the relative susceptibility of the strains to drugs in vitro. Regarding metronidazole, tinidazole and satranidazole susceptibility, different degrees of susceptibility were determined. However, no difference in nitazoxanide susceptibility was found between the clonal strain tested and a reference strain. Conclusions This is the first description of biological variability in clonal stock of Trichomonas vaginalis. Different degrees of drug susceptibility were determined among clonal strains tested. Further studies will be necessary to ascertain the importance of this variability in clinical infection.

58-60

Authors: Mohammad Khalili, Ehsanollah Sakhaee, Mohammad Reza Aflatoonian, Naser Shahabi-Nejad

Objective To determine the prevalence of Coxiella burnetii (C. burnetii) antibody positive randomly selected dairy herds in southeast Iran (Kerman). Methods Bulk tank milk samples were collected randomly from 44 sufficiently large commercial dairy herds, included near 12 000 dairy cattle, in Kerman (The largest province of Iran), southeast Iran. The samples were tested for antibodies against C. burnetii using the commercial CHEKIT® Q fever antibody ELISA Test Kit (Idexx, Liebefeld-Bern, Switzerland). Results The prevalence of positive, negative and intermediate herds were 45.4%, 43.2% and 11.4%, respectively. Conclusions The result supports the hypothesis of high prevalence and endemic pattern of Q fever in Iran. This investigation highlights the importance of further studies on Q fever in Iran.

64-66

Authors: Haris M Khan, Fatima Shujatullah, Mohammad Ashfaq, Adil Raza

Objective: To determine the prevalence of malaria in Aligarh and analyze species dominance in different years over a decade. Methods: Diagnosis of malaria was done using microscopy as gold standard, rapid antigen detection assays and quantitative buffy coat (QBC) assays. Giemsa stained blood smear examination was done, thick and thin films were examined for presence of different Plasmodium spp. Rapid antigen detection assays employing detection of HRP-2 and parasite lactate dehydrogenase antigen (pLDH) by immunochromatography was done in patients whose blood smear found to be negative by conventional Giemsa slide examination. QBC was done in cases where there is strong clinical suspicion of malaria with blood smear negative, in patients with chronic malaria, splenomegaly, or in those patients who had inadequate treatment and for post-treatment follow up. Results: Plasmodium vivax and Plasmodium falciparum were only species detected in our hospital. Overall prevalence of malaria in Aligarh was found to be 8.8%. The maximum prevalence of 20.1% was observed in year 2008 and lowest 2.3% in 2002. Conclusions: High prevalence of malaria is observed in this part of country with dominance of both species particularly Plasmodium falciparum should be monitored and factors accounting for occurrence should be studied to employ effective control measures.

72-75

Authors: Sreenivasan Sasidharan, Subramanion Jo Thy Lachumy, Manickam Ravichandran, Lachimanan Yoga Latha, Surasa Rao Surya Gegu

Objective To study the epidemiology of Helicobacter pylori (H. pylori) infection according to age group. Methods H. pylori infection data among 1 965 consecutive patients referred to the Endoscopy Unit collected at Sungai Petani Hospital for oesophagogastro-duodenoscopy (OGD). The patients were divided into 9 age groups (10–19, 20–29, 30–39, 40–49, 50–59, 60–69, 70–79, 80–89 and 90–99 years). In addition these groups were further divided into three minor group namely young adults (10–39), older adults (40–69) and geriatric groups (70–99). Results Overall prevalence of infection of H. pylori was analyzed and found that the prevalence increase with age (P<0.05). When the patients divided by ethnic and gender group with age, prevalence rate among young adults and older adults significantly higher (P<0.05) compared to geriatric groups across all races and gender (P<0.05). Furthermore, significantly higher number of males were infected compared to female (P<0.05) but such trend was only observed among older adult groups. In addition, there is a significant differences in H. pylori infection prevalence rates among ethnic groups (highest in Indians adults, followed Chinese and low in Malays, P<0.05). Conclusions The overall prevalence of H. pylori did increase with age group across ethnicity and gender, in Northern Peninsular Malaysia.

79-80

Authors: Rupashree Sundar, Lakshmi Rao, Geetha Vasudevan, Pruthvi Baise Chandrappa Gowda, Rau Neeleshwar Radhakrishna

Cryptococcus neoformans has been recognized as a human pathogen over centuries. This has achieved new prominence in the recent years as it is an opportunistic fungi causing fatal, deep mycotic infections in immunocompromised states. Although cryptococcus is principally a pathogen of central nervous system, wide variety of other organs may also be involved. Gastrointestinal cryptococcosis is rarely reported either as an isolated finding or in a disseminated disease. However, even with the strikingly increased incidence of the disease, occurrence of obvious gastrointestinal symptoms directly attributable to cryptococcosis is outstandingly rare. We report a case of gastric cryptococcal infection with esophageal herpes as an initial presentation in an AIDS patient.

83-84

Authors: Peter George

To leptospirosis is the commonest spirocheatal infection in the tropical and temperate countries of Indian sub-continent and Africa and the most common zoonosis worldwide. The protean manifestation of this infectious disease is a challenge for practising clinicians across the world. In poor developing countries, at most clinical suspicion it is essential in the diagnosis of this disease. In this report, we are able to document two uncommon manifestations of leptospirosis, namely Sweet's syndrome and central nervous system vasculitis.

925-929

Authors: Yongxiang Zhao, Jing Tang, Qin Yao, Yuan Zhou, Huange Zhao, Xiaoyun Zeng, Jiaqi Shi, Guorong Luo, Xiaoxun Xie, Sufang Zhou, Zuguo Liu, Xiaoling Lu, Donghai Lin, Jianming Liu

Objective To investigate the effect of fusion proteins expressed by the fused gene of porcine α 1, 3 galactosyltransferase (α1, 3 GT) and enhanced green fluorescent protein (EGFP) on the green fluorescence intensity of EGFP. Methods The fragment containing α 1, 3GT was firstly recovered after the pcDNA3.1- α 1, 3GT recombinant vector were digested with Bam HI and Eco RI, and then, the resultant fragment was ligated to the pEGFP-N1 vector which was also digested with the same enzymes. The new recombinant eukaryotic expression pEGFP/α 1, 3GT vector was obtained and sequenced. The pEGFP/α 1, 3GT was used to transfect human lung carcinoma cells A549 and HEKC 293FT, and the expression of EGFP was quantitatively analyzed by fluorescent microscope and flow cytometry. Results The positive percentage of A549 was 80.5%, and that of 293 FT was 86.5% 48 hours after the two cell lines both were transfected by pEGFP-N1. The positive percentage of A549 was 75.8%, and that of 293 FT was 81.2% 48 hours after the two cell lines were transfected by pEGFP/α 1, 3GT. The mean fluorescence intensities of A549 transfected with pEGFP-N1 and pEGFP/α 1, 3GT were 1.21 and 0.956, respectively when compared with that of A549 without transfection. Meanwhile, the those of the 293FT that were transfected with pEGFP-N1 and pEGFP/α 1, 3GT were 7.66 and 1.00, respectively when compared with that of 293FT cells without transfection. Conclusions These results suggested that the expression of EGFP gene fused with porcine α 1, 3GT gene was partly inhibited.

934-938

Authors: Mauro Prato, Valentina Gallo, Elena Valente, Amina Khadjavi, Giorgia Mandili, Giuliana Giribaldi

Objective To investigate the effect of malarial pigment (hemozoin, HZ) on expression of heat shock proteins (HSPs) and cell viability in human monocytes by using a stable cell line (THP-1 cells). Methods THP-1 cells were fed with native HZ or treated with pro-apoptotic molecule gliotoxin for 9 h. Thereafter, the protein expression of HSP-27 and HSP-70 was evaluated by western blotting. Alternatively, HZ-fed cells were cultured up to 72 h and cell viability parameters (survival, apoptosis and necrosis rates) were measured by flow cytometric analysis. Results HZ increased basal protein levels of HSP-27 without altering those of HSP-70 in THP-1 cells, and promoted long-term cell survival without inducing apoptosis. As expected, gliotoxin inhibited HSP-27 protein expression and promoted long-term cell apoptosis. Conclusions Present data show that HZ prevents cell apoptosis and enhances the expression of anti-apoptotic HSP-27 in THP-1 cells, confirming the previous evidences obtained from HZ-fed immunopurified monocytes. Since the use of a stable cell line is pivotal to perform HSP-27 silencing experiments, monocytic THP-1 cells could be a good candidate line for such an approach, which is heavily required to clarify the role of HSP-27 in survival of impaired HZ-fed monocytes during falciparum malaria.

943-947

Authors: Ukwe Chinwe V, Ekwunife Obinna I, Epueke Ebele A, Ubaka Chukwuemeka M

Objective To determine the suppressive and curative activity of aqueous leaf extract of Ageratum conyzoides (A. conyzoides) in combination with chloroquine and artesunate, respectively against Plasmodium berghei infection in mice. Methods Using malaria (Plasmodium berghei) infected albino mice of both sexes, aqueous extracts of A. conyzoides in combination with chloroquine and artesunate were tested for antimalarial activity, respectively. Four-day suppressive test and Rane's curative test were carried out. Results Suppressive tests showed significant dose dependent reduction in parasitemia level produced by the extract-chloroquine and extract-artesunate combinations. Suppressive activities of both extract-drug combinations were greater than the individual drugs alone. Extract-chloroquine (100:5) produced the highest suppressive effect (98% suppression). Curative tests showed absolute survival in two extract-drug combinations. Two extract-drug combinations produced higher curative effects than the individual drugs alone. The highest dose combinations of extract-chloroquine (100:5) and extract-artesunate (100:5) produced absolute parasitemia clearance (cure) in the infected mice. Conclusions The study indicated that aqueous extract of A. conyzoides had the ability to potentiate the antimalarial activity of chloroquine and artesunate against induced plasmodiasis in mice. It contributes a lot in the malaria endemic and poverty stricken tropics.

954-956

Authors: Supratik Chakraborty, Someshwar Singha, Goutam Chandra

Objective To observe the effect of two common organic acids on the larvae of Aedes aegypti (Ae. aegypti) (L), the natural vector of dengue fever/dengue hemorrhage fever, chikugunya and allergic skin reaction especially in children. Methods Two common organic acids (lactic acid and orthophosporic acid of gradually increasing concentration) were used against laboratory reared third instars larvae of Ae. aegypti in order to observe the rate of mortality after 8, 16 and 24 h of post exposure respectively in laboratory. Results Larval mortality rates recorded were in the following sequences: orthophosphoric acid and lactic acid at 1:1 combination >orthophosphoric acid>lactic acid. Conclusions These two organic acids may be used perfectly in combination (1:1) along with other conventional vector control methods to reduce the Ae. aegypti population, especially in those areas where surveillance and supervisory mechanism are poor or insufficient.

961-964

Authors: Shyamapada Mandal, Manisha DebMandal, Nishith Kumar Pal, Krishnendu Saha

Objective To ascertain the potential antibacterial activity of honey against clinical isolates of Escherichia coli (E. coli), Pseudomonas aeruginosa (P. aeruginosa) and Salmonella enterica serovar Typhi (S. enterica serovar Typhi) by in vitro methods. Methods The partial inhibitory concentration (PIC), minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) values of the autoclaved honey (extracted from Apis indica hive by indigenous method) were determined for S. enterica serovar Typhi (n=8; from blood cultute), E. coli (n=5; from urine culture) and P. aeruginosa (n=5; from pus culture) isolates by in vitro methods. Results The PICs of the honey tested for the isolates ranged 0.50%-1.25 % (v/v) for S. enterica serovar Typhi, 0.75%-1.50% (v/v) for E. coli and 1.00%-1.25 % (v/v) for P. aeruginosa, while the MICs ranged 1.75%–3.00% (v/v), 3.00%-3.50% (v/v) and 3.50% (v/v), respectively. The P. aeruginosa and E. coli isolates had MBC value of 4.00% (v/v); the S. enterica serovar Typhi showed MBCs in between 3.00% and 3.50% (v/v). The bactericidal activity of honey was achieved at concentration 3.00% (v/v) for S. enterica serovar Typhi and E. coli, and at 3.50% (v/v) for P. aeruginosa. Conclusions The excellent antibacterial activity of honey against clinical bacterial isolates indicates the usefulness of honey in clinical practice against bacterial infection.

971-976

Authors: Evan Prince Sabina, Annie Deborah Harris Souriyan, Deborah Jackline, Mahaboob Khan Rasool

Objective To explore the hepatoprotective and antioxidant effects of piperine against acetaminophen-induced hepatotoxicity in mice. Methods In mice, hepatotoxicity was induced by a single dose of acetaminophen (900 mg/kg b.w. i.p.). Piperine (25 mg/kg b.w. i.p.) and standard drug silymarin (25 mg/kg b.w. i.p.) were given to mice, 30 min after the single injection of acetaminophen. After 4 h, the mice were decapitated. Activities of liver marker enzymes [(aspartate transaminase (AST), alanine transaminase (ALT), and alkaline phosphatase (ALP)] and inflammatory mediator tumour necrosis factor-alpha (TNF-α) were estimated in serum, while lipid peroxidation and antioxidant status (superoxide dismutase, catalase, glutathione peroxidase, glutathione reductase, glutathione-s-transferase and glutathione) were determined in liver homogenate of control and experimental mice. Results Acetaminophen induction (900 mg/kg b.w. i.p.) significantly increased the levels of liver marker enzymes, TNF-α, and lipid peroxidation, and caused the depletion of antioxidant status. Piperine and silymarin treatment to acetaminophen challenged mice resulted in decreased liver marker enzymes activity, TNF-α and lipid peroxidation levels with increase in antioxidant status. Conclusions The results clearly demonstrate that piperine shows promising hepatoprotective effect as comparable to standard drug silymarin.

977-981

Authors: Subramanian Kumaran, Balaraman Deivasigamani, Kumarappan Alagappan, Mannikam Sakthivel, Rajamani Karthikeyan

Objective To determine the prevalence and antibiotic resistance of Escherichia coli (E. coli), in seafood obtained from Cuddalore and Parangipettai fish landing centres. Also, to identify the susceptibility of E. coli against predominant seaweeds red alga Kappaphycus alvarezii (K. alvarezii) and brown alga Padina boergessenii (P. boergessenii) extracts as sulfated polysaccharides and polyphenols respectively. Methods A total of 48 samples (Two stations Cuddalore and Parangipettai, Tamil Nadu, India). Sampling area are fish landing centre where fishes caught from sea and estuary, seafood processing plants (packing and ice packed fishes) and local fish markets (fish samples). After isolation totally 80 strains were analyzed for its antimicrobial resistance and sensitivity against commercially 10 antibiotics. The ampicillin resistant E. coli CE21 was identified through molecular techniques as 16S rDNA sequencing. Two seaweeds K. alvarezii and P. boergessenii were screened for antibacterial activity against 12 antibiotic resistant E. coli strains. Results Totally 48 swabbed samples from two different fish handling area were characterized for total bacterial and E. coli count. Mostly, the E. coli strains were isolated from fish local market and seafood processing plants before and after packaging process. In that maximum 56.25% strains were resistant to ampicillin and the minimum 2.5% strains were resistant to chloramphenicol. Therefore, the E. coli CE21 was identified through molecular techniques E. coli (GenBank accession number GU065251), The MIC value for polyphenol extract was slightly less than sulfated polysaccharides. E. coli strain isolated from Parangipettai was considerably increased MIC value that Cuddalore. Conclusions The polyphenol and sulfated polysaccharides showed promising inhibitory response against all antimicrobial resistant E. coli strains and in particular the inhibitory response of ampicillin resistant E. coli.

988-992

Authors: Mohammed Haneefa KP, Shahima Hanan K, Saraswathi R, Guru Prasad Mohanta, Chandini Nayar

Objective To formulate Pothos scandens Linn (P. scandens) leaf extract in to a gel and investigate their burn wound healing activity. Methods Ethanolic extract of dried leaves of P. scandens were subjected to priliminery phytochemical evaluation and wound healing activities studies. Different gel formulations of ethanolic extract of P. scandens (4% w/v) were prepared using polymers carbopol 934 and carbopol 940 by varying their concentration. These formulations were evaluated for physical parameters, drug content, pH, viscosity, extrudability, spreadability, primary skin irritation, pharmacological activity and stability. Results Wound healing studies of ethanolic extract revealed that P. scandens treated animals were found to epithelise in 22 days while the solvent control and the untreated rats epithelise within 35 and 40 days respectively. The formulation with 1.5% w/w carbopol 934 was found to be more promising as it shows better physicochemical characteristics, higher pharmacological activity and stability compared to other formulations. Conclusions P. scandens alcoholic extract shows significant improvement in burn wound contraction and hence this is a promising candidate in burn wound healing.

997-1000

Authors: Kuldeep Kumar, Pankaj Kumar Singh, Juhi Tomar, Swati Baijal

Dengue fever is a mosquito born viral infection, and the complicated form of dengue is dengue hemorrhagic fever (DHF). In the recent decades incidence and distribution of dengue has increased dramatically. Dengue viruses belong to family flaviviridae with four serotypes and are transmitted mainly by mosquito Aedes aegypti. Today almost two-fifth of world's population (2.5 million) is at risk of dengue and no specific antiviral drug or vaccine is available against it. Uncontrolled population growth in Africa and South East Asia has increased number of susceptible hosts in urban and semi urban areas. About 40% of world population resides in the high risk area for dengue transmission. According to latest estimates by WHO, yearly 50 to 100 million infections occur globally, this includes around 500 000 DHF and 22 000 deaths, mostly among children. Only symptomatic treatment in the form of analgesic, antipyretics and body fluid management is provided to the patient. Prevention strategies mainly focus on two approaches, firstly on activities to control vector and secondly on activities to protect human from mosquito bite but there is always concerns regarding their sustainably and effectiveness. Theoretically development of an effective dengue vaccine is feasible and production of an effective and affordable vaccine could be a viable option to save humans from this dreadful disease. Conceptually vaccine production is possible, but it has to be tetravalent, providing immunity against all serotypes. Few candidate vaccines are in advance stage of their development; however international cooperation is needed to make these vaccines available on cheaper rates to the poor and vulnerable countries. Objective of this review is to discuss various aspects related to dengue, its epidemiology, available preventive methods, need for vaccine and challenges in its development.

1004-1005

Authors: Sumeet Bhargava, Rajul Rastogi, Gaurav Jindal, Amrit Kumar Singh, Vaibhav Rastogi

Tuberculous infection manifesting as an isolated intramedullary tuberculoma of the spinal cord is distinctly unusual. We report a case of a 35 year old woman presenting with an insidious onset of myelopathy, where MRI showed characteristic imaging findings suggestive of intramedullary tuberculosis.

841-845

Authors: Sedaghat Mohamad Medhi, SaneiAli Reza, Khnavi Mahnaz, Abai Mohammad Reza, Hadjiakhoondi Abbas, Mohtarami Fatemeh, Vatandoost Hassan

Objective To determine phytochemistry and larvicidal activity of Eucalyptus camaldulensis against Anopheles stephensi. Methods The chemical compositions of the leaf essential oils were analyzed using gas chromatography/mass spectrometry (GC/MS). The larvicidal activity of essential oils and extract of leaf were tested against 4th instar larvae of laboratory-reared Anopheles stephensi. Results GC/MS analyses identified the presence of 28 compounds corresponding to 99.60% of the total oil. The main constituents in the leaf essential oil were 1,8-cineole (69.46%), γ –Terpinene (15.10%), α –Pinene (5.47%) and Globulol (2%). The leaf extract and volatile oil exerted significant larvicidal activity with LC50 values of 89.85 and 397.75 ppm, respectively. Clear dose-response relationships were established with the highest dose of 320 ppm essential oil extract resulted almost 100% mortality in the population. Conclusion The larvicidal properties suggest that the essential oil of plant is a potential source of valuable larvicidal compounds against malaria vector and can be used as an alternative to synthetic insecticides.

851-854

Authors: Lijuan Zhang, Daxin Ni, Zijian Feng

Objective To report a training course on the laboratory diagnoses of rickettsioses that 10 provincial/city CDCs participated in laboratory external quality assurance (EQA) based on the serological specific antibodies detection and rapid PCR amplifying targeted genes of rickettsiae. Methods An EQA program to evaluate the following laboratory procedures was developed to detect rickettsiae: (1) immunofluorescent assay (IFA) to detect specific antibodies of A. phagocytophilum, R. heilongjiangensis and O. tsutsugamshi respectively.(2) Two sets of nested PCR were used amplifying groEL genes for most members of the family Rickettsiaceae and amplifying 16SrRNA genes for the most members of family anaplasmae, respectively. A scoring scheme based on the distribution of the median antibody titer values of the serologic assays was designed and a ranking list of the scores of the PCR results based on the detected minimal copy numbers of reference DNA was created. Results Among nine laboratories who reported the results on time, eight laboratories gave acceptable serologic results, the other one provided an unacceptable antibody titer (1:2 vs median 1:64) results for O. tsutsugamshi. The limits of detection (LOD) for the PCR amplifying for five references DNA ranged from 1copy/μ L to 106 copy/μ L. Conclusion We successfully trained and popularized modern diagnostic methods of rickettsiae in 10 provincial CDCs in China and first conducted the EQA projects and evaluated the results.

860-865

Authors: CVM Naresh Kumar, P Sangamithra, M Rajasekhar, DVR Saigopal

Objective The study involved survey and screening of areas suspected of chikungunya virus (CHIKV) infection, characterizing the causative agent and identifying the circulating CHIKV genotype. Methods Acute phase samples were screened by the use of RTPCR using primer set DVRChk-F/DVRChk-R whereas convalescent samples were tested by CHIKV IgM strips. Results Two hundred and seventy five acute phase samples were screened by RT-PCR, of which 149(54.18%) showed positivity for CHIKV. Later on 192 convalescent phase samples were tested for CHIKV specific antibodies in which 125(65.10%) samples were found to be positive. Four CHIKV strains were selected and subjected to cloning followed by nucleotide sequencing and were submitted to the Genbank DNA database with the Accession numbers (GQ119362, GQ119363, GQ119364, and FJ225403). The Sequence analysis of “CHIK-Kadapa” strain (GQ119362) with other CHIKV isolates suggested that the present CHIKV strain has (99.23±0.52) % and 100 % identity with Central East South African isolates (CESA) at nucleotide and amino acid levels respectively. Two unique non synonymous mutations S168L and D183V were depicted in E1 gene of the selected strains of the present study. Conclusion The 14 months survey revealed the circulation of CHIKV in 2008-2009 in Andhra Pradesh and the causative agent is identified to be of Central East South African (CESA) origin. The importance of the non synonymous mutations (S168L and D183V) and their role in the mobility and strength of the E1-E1 and E1-E2 interactions needs further investigations. The study also urges the need for intensifying the epidemiological and entomological surveillance to combat any such CHIKV outbreak in the near future.

871-873

Authors: S Kumar, V Kumar, Om Prakash

Objective To investigate the antidiabetic and anti-lipemic effects of Cassia siamea methanolic leaves extract. Methods The antidiabetic study was performed by measuring blood glucose level with elegance glucometer at weekly intervals i.e. 0, 7, 14 and 21 in normal and streptozotocin induced diabetic rats. Total cholesterol, triglyceride and HDL-cholesterol were determined in normal and streptozotocin induces diabetic rats by autoanalyser. Glibenclamide was used as a reference drug at a dose of 10 mg/kg. Results After the oral administration of extracts at doses of 250 mg/kg and 500 mg/kg for three weeks, blood glucose levels and body weights were significantly improved (P<0.01). Daily oral treatment with the extract also resulted in significantly reduction of serum cholesterol and triglycerides. HDL-cholesterol level was found to be improved to (P<0.01). Conclusion The Cassia siamea leaf extract is useful in controlling blood glucose level as well as improving lipid metabolism and body weight in rats with induced diabetes.

878-883

Authors: Abdul Abduz Zahir, Abdul Abdul Rahuman, Asokan Ba-gavan, Gandhi Elango, Chinnaperumal Kamaraj

Objective To determine the adult emergence inhibition (El) and adulticidal activities of hexane, chloroform, ethyl acetate, and acetone leaves extracts of Anisomeles malabarica (A. malabarica), Euphorbia hirta (E. hirta), Ocimum basilicum (0. basilicum), Ricinus communis (R. communis), Solanum trilobatum (S. trilobatum), Tridax procumbens (T. procumbens) and seeds of Gloriosa superba (G. superba) against Anopheles stephensi (An. stephensi). Methods The El and adulticidal trials were carried out according to World Health Organization (WHO) procedures with slight modifications. The extracts were diluted in dimethyl sulphoxide in order to prepare a serial dilution of test dosages (15.625, 31.25, 62.5, 125, 250, 500 and 1 000 jU g/mL). Five duplicate trials were carried out for every sample concentration, and for each trial a negative control was included and the mortality was determined after 24 h of exposure. Results The highest El activity was found in ethyl acetate extracts of A. malabarica, chloroform extracts of 0. basilicum, S. trilobatum, acetone of extract of R. communis, T. procumbens, and seed extract of G. superba with EI50 values 143.12, 119.82, 157.87, 139.39, 111.19, and 134.85 fig/mL, and the effective adulticidal activity was observed in chloroform, acetone extracts of G. superba, T. procumbens, R. communis, S.trilobatum and ethyl acetate extract of 0. basilicum with LD50 values 120.17, 108.77, 127.22, 163.11, 118.27, and 93.02 l-i g/mL, respectively. Chi-square value was significant atP<0.05 level. Conclusion These results should encourage further efforts to investigate the compounds that might possess good El and adulticidal properties when isolated in pure form.

887-889

Authors: CF Oguejiofor, IS Ochiogu, CJ Umeoduagu

Objective To investigate the effects of comparatively high doses of diminazene aceturate on the reproductive performance of female rats in the early stage of pregnancy. Methods After oestrus synchronisation and successful mating, 20 pregnant female rats were randomly divided into four groups (A-D). Group A rats served as the control and were given single intraperitoneal injection of 0.5 mL sterile water (vehicle only) while groups B, C and D rats were given single intraperitoneal doses of 7, 14 and 21 mg/kg body weight diminazene aceturate respectively, on day 7 of pregnancy. The gestation length, litter size and weight at birth, and areas of foetal résorption in the uterus were determined post partum. The post-implantation survival index (%) and the gestation index (group %) were also evaluated for rats in all the groups. Results There was a graded increase in the number of observed resorbed foetuses as the dose of diminazene aceturate was increased, although only groups C (14 mg/kg) and D (21 mg/kg) revealed a significant decrease (P<0.01, ANOVA) in the post implantation survival index of rat embryos. There was also a significant decrease (P<0.05) in the litter weights of rats in groups C and D. Conclusion Although the pregnant rats showed no overt signs of systemic toxicity even at the highest dose of 21 mg/kg body weight diminazene aceturate in this study, it was concluded that the use of high doses of diminazene aceturate in an effort to combat resistant trypanosomes could have adverse reproductive effects on female animals in the early period of pregnancy.

894-897

Authors: M Johnson, EG Wesely, MI Zahir Hussain, N Selvan

Objective To evaluate the phytochemical and anti-bacterial potential of mother plants in vivo and in vitro derived callus of Baliospermum montanum (B. montanum) (Willd.) Muell.-Arg. leaves and root. Methods The in vitro derived rootlets and leaves segments of B. montanum were cut into 0.5-0.7 cm in length and cultured on Murashige and Skoog solid medium supplemented with 3% sucrose, gelled with 0.7% agar and different concentration of 2, 4-D either alone or in combinations. The preliminary phytochemical screening was performed by Harborne method. Antibacterial efficacy was performed by well diffusion method and incubated for 24 h at 37°C. Results The highest percentage of callus formation (leaves segments 86.9±0.56; root segments 78.7±0.51) was obtained on Murashige and Skoog's basal medium supplemented with 3% sucrose and 2.0 mg/L of 2,4-Dichlorophenoxy acetic acid. The phytochemical study revealed the high quantity presence of steroids, triterpenoids, glycosides, saponins, alkaloids, flavanoids, phenolic compounds, tannins, sugars etc of root and leaves derived calli. The ethanol extract of leaves segment derived calli of B. montanum showed the maximum solubility and antimicrobial activity with the MIC ranged from 100 to 200 μL. Conclusion The preliminary phytochemical study confirmed that the calli mediated tissues showed the higher percentage of metabolite constituents and extraction value compared to the in vivo leaves and roots. The present study observation suggested that a possibility to establish high yielding genotypes by in vitro culture for production of medicinally important bioactive compounds.

902-905

Authors: HS Wijewantha, KH Sellahewa

Objective To identify the clinical manifestations of hump nosed viper envenomation and to recognize the available treatment methods to prevent complications. Methods Descriptive observational study involving a series of 1 543 patients admitted with hump nosed viper bite to 5 major hospitals in Sri Lanka was conducted. Data collection was done consecutively during February 1990 and February 2008. Except Hypnale, identification of the biting snake was made by the corresponding author after visual examination of the dead or live snakes, which were brought to hospital. Results Sixty seven (4.34%) patients developed systemic effects and two (0.1%) patients died due to effects of envenomation or complications of treatment. Systemic effects varied from coagulopathy, nephropathy to some neurological manifestations. Fifty nine (3.8%) patients had only coagulopathy and they received either, intravenous isotonic saline to ensure adequate urine out put i.e. 0.5 mL/kg /hour or 15 mL/kg of fresh frozen plasma (FFP). None of the patients that had coagulopathy developed renal failure. Contamination of the sample by mildly venomous species of Hypnale may have contributed to the low incidence of systemic complications. Conclusion It is likely that early hydration with normal saline or FFP can prevent acute renal failure. FFP showed a tendency for early correction of coagulopathy. Role of FFP in hump nosed viper envenomation is worth studying in randomized double blind controlled clinical trials.

913-916

Authors: Peng Kong, Yan Kong, Xu Jiang, Xiaonua Wang

This article introduced the vision building concept about human influenza pandemic prevention and control. Different visions were built by creating different shapes of building blocks which also represented different organizations and physical facilities, respectively. The around-view reflection is required to be developed in the process of building so as to search for the ideal pattern. The correlation of all sectors and systems are established to combine different kinds of things, from one family to another, from communities, towns, counties, cities, rural areas, provinces to the state to handle trivial problems. These training objectives have been successfully accomplished, which has not only enriched the knowledge about prevention and control of influenza pandemic between different departments but also clarified the roles and responsibility. It lays the firm foundation for next cooperation between different departments, and make a bridge for the objective and choice of channel over human influenza pandemic prevention and control.

922-924

Authors: G Peter, SR Basti, AS Joy

Multiple abscesses of the costo-chondral junctions are very uncommon in practice. In this report we present the case of a 55 year old man who presented to us with chest pain and fever of few months duration. On imaging with ultrasound and CECT we were able to demonstrate multiple abscesses of costo-chondral junctions bilaterally. We confirmed tuberculosis by FNAC and BACTEC cultures from abscesses.