Asian Pacific Journal of Tropical Medicine

673-677

Authors: Raewadee Wisedpanichkij, Wanna Chaicharoenkul, Poonuch Mahamad, Prapichaya Prompradit, Kesara Na-Bangchang

Objective To investigate the association between amplification of the two regulatory genes controlling glutathione (GSH) levels, glutathione reductase (PfGR) and glutathione S-transferase (PfGST) genes and sensitivity of Plasmodium falciparum (P. falciparum) isolates collected from different malaria endemic areas of Thailand to standard antimalarial drugs. Methods A total of 70 P. falciparum isolates were collected from endemic areas of multi-drug resistance (Tak, Chantaburi and Ranong Provinces) during the year 2008-2009. The in vitro assessment of antimalarial activity of P. falciparum clones (K1- and Dd 2 chloroquine resistant and 3D7-chloroquine sensitive) and isolates to chloroquine, quinine, mefloquine and arteusnate was performed based on SYBR Green modified assay. Results 68 (97.14%), 11 (15.71%) and 28 (40%) isolates respectively were classified as chloroquine-, quinine- and mefloquine-resistant isolates. With this limited number of P. falciparum isolates included in the analysis, no significant association between amplification of PfGST gene and sensitivity of the parasite to chloroquine, quinine, mefloquine and quinine was found. Based on PCR analysis, Dd2, K1 and 3D7 clones all contained only one copy of the PfGST gene. All isolates (70) also carried only one copy number of PfGST gene. There appears to be an association between amplification of PfGR gene and chloroquine resistance. The 3D7 and Dd2 clones were found to carry only one PfGR gene copy, whereas the K1 clone carried two gene copies. Conclusions Chloroquine resistance is likely to be a consequence of multi-factors and enzymes in the GSH system may be partly involved. Larger number of parasite isolates are required to increase power of the hypothesis testing in order to confirm the involvement of both genes as well as other genes implicated in glutathione metabolism in conferring chloroquine resistance.

682-686

Authors: Anjana Goel, Sandeep Kumar, Ashok Kumar Bhatia

Objective To investigate the protective role of Ocimum sanctum (O. sanctum) leaves against Salmonella typhimurium(S. typhimurium) infection in rats by inducing TNF-α, IFN-γ& IL-2 cytokines. Methods Wistar albino rats were fed with aqueous extract of O. sanctum leaves using 250 mg/kg body weight dose once a day for 20 consecutive days. Control rats were fed with placebo. Rats were infected with LD 50 dose of S. typhimurium infection and monitored for their survival. Bacterial blood burden in both the groups was compared and numbers of activated peritoneal macrophages were counted. Concentration of TNF-α, IFN-γ and IL-2 cytokines in serum during different time intervals was assayed by sandwich ELISA. Results Rats of control group showed a high mortality rate and had higher bacterial blood burden when compared with O. sanctum extract fed rats. There was a significant increase in the number of S. typhimurium engulfed peritoneal macrophages in the peritoneal fluid of O. sanctum fed animals. The protective control against bacterial infection in O. sanctum fed rats was associated with elevated level of TNF-α, IFN-γ and IL-2 cytokines in serum. Conclusions These findings suggest that orally administered O. sanctum leaves extract effectively enhanced activation in macrophage and lymphocytes, depicted by the elevated serum concentration of TNF-α, IFN-γ and IL-2 cytokines, leading to induce a protective resistance against Salmonella typhimurium infection.

691-695

Authors: Marimuthu Govindarajan

Objective To determine the larvicidal and repellent activities of Sida acuta Burm. F. (Family: Malvaceae)extract against Culex quinquefasciatus, Aedes aegypti and Anopheles stephensi. Methods Twenty five late III instar larve of three mosquito species were exposed to various concentrations (15-90 mg/L) and were assayed in the laboratory by using the protocol of WHO 2005; the 24 h L C 50 values of the Sida acuta leaf extract was determined following Probit analysis. The repellent efficacy was determined against three mosquito species at three concentrations viz., 1.0, 2.5 and 5.0 mg/cm 2 under the laboratory conditions. Results Results showed varying degree of larvicidal activity of crude extract of Sida acuta against three important mosquitoes with LC 50 values ranging between 38 to 48 mg/L. The crude extract had strong repellent action against three species of mosquitoes as it provided 100% protection against Anopheles stephensi for 180 min followed by Aedes aegypti (150 min) and Culex quinquefasciatus (120 min). Conclusions From the results it can be concluded the crude extract of Sida acuta was an excellent potential for controlling Culex quinquefasciatus, Aedes aegypti and Anopleles stephensi mosquitoes.

702-706

Authors: Bagepalli Srinivas Ashok Kumar, Kuruba Lakshman, KN Jayaveera, Devangam Sheshadri Shekar, Avalakondarayappa Arun Kumar, Bachappa Manoj

Objective Methanolic extract of Amaranthus spinosus (A. spinosus) leaves was screened for antioxidant and antipyretic activities. Methods Antioxidant activity was measured by 1,1-diphenyl-2-picryl-hydrazile (DPPH) free radical scavenging, superoxide anion radical scavenging, hydroxyl free radical scavenging, nitric oxide radical scavenging, 2,2 '-azinobis-3-ethylbenzothiazole-6-sulfonic acid (ABTS) radical scavenging assays and total phenolic content was also determined. Antipyretic activity of methanolic extract of A. spinosus was measured by yeast induced pyrexia method at concentration of 200 and 400 mg/kg using paracetamol as standard drug. Results Methanolic extract of A. spinosus showed potent antioxidant activity. The IC 50 value was (87.50 ±3.52) μg/mL, (98.80±1.40) μg/mL, (106.25±0.20) μg/mL, (88.70±0.62) μg/mL and (147.50±2.61) μg/mL for DPPH, superoxide, hydroxyl, nitric oxide and ABTS radical scavenging activities. Methanolic extract of A. spinosus showed significant (P <0.01) antipyretic activity.

711-714

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

Objective To evaluate the antibacterial activity of Emblica officinalis Gaertn (E. officinalis; Family: Euphorbiaceae) seed and Nymphae odorata Aiton (N. odorata; Family: Nymphaeaceae) stamen extracts, alone and in combination, and in combination with amoxicillin (Ax) against Staphylococcus aureus (S. aureus). Methods Antibacterial activity of ethanolic extracts of amla, E. officinalis, seed (AMS; 500 μg) and sapla, N. odorata, stamen (SAP; 500 μg) for 12 methicillin-resistant S. aureus (MRSA) isolates was determined following agar diffusion; in order to assess the combined antibacterial activity, AMS (250 μg) plus SAP (250 μg) were considered. The Ax (10 μg) activity alone and in combination with AMS (250 μg), and SAP (250 μg) was determined by disk diffusion. The zone diameters of inhibition (ZDI s) for the agents were recorded, and growth inhibitory indices (GII s) were calculated. Results The MRSA isolates (n = 12) had AMS (500 μg) and SAP (500 μg) ZDI s of 12-19 mm and 21-24 mm, respectively. The ZDI s (range 24-27 mm) increased by 3-4 mm due to combined action of AMS (250 μg) and SAP (250 μg) indicating synergy between extracts for MRSA (GII 0.634-0.742). The MRSA isolates were resistant to Ax (ZDI: 8-11 mm), which in combination with AMS and SAP had synergistic effect, both due to increased ZDI [mean± SD = (3.5 ± 0.577) mm] and GII (0.631-0.894). Conclusions The data suggest that the plants, E. officinalis and N. odorata alone or in combination, are promising in the development of phytomedicines, which may be used, alone or in combination with the antibiotic, Ax, against MRSA infection.

723-726

Authors: Selvamaleeswaran Ponnusamy, Wesely Ebenezer Gnanaraj, Johnson Marimuthu @Antonisamy, Velusamy Selvakumar, Jeyakumar Nelson

Objective To investigate the antimicrobial activity of Clitoria ternatea(C. ternatea) against the fish pathogens viz., Pseudomonas aeruginosa(P. aeruginosa), Escherichia coli(E. coli), Klebsiella pneumonia(K. pneumonia), Bacillus subtilis(B. subtilis), Aeromonas formican(A. formicans)s, Aeromonas hydrophila(A. hydrophila) and Streptococcus agalactiae(S. agalactiae) isolated from diseased Tilapia (Oreochromis niloticus). Methods The extracts of C. ternatea was tested against P. aeruginosa, E. coli, K. pneumonia, B. subtilis, A. formicans, A. hydrophila and S. agalactiae by the agar well diffusion method. Results Different extracts of C. ternatea showed inhibitory effects against P. aeruginosa, E. coli, K. pneumonia, B. subtilis, A. formicans, A. hydrophila and S. agalactiae. Ethyl acetate extracts of C. ternatea showed maximum of zone of inhibition against A. formicans (18 mm), A. hydrophilia (19 mm), B. subtilis (19 mm) and P. aeruginosa (21 mm) next to that ethanol extract of C. ternatea showed A. formicans (18 mm) and E. coli (14 mm) followed by Acetone extract showed maximum zone of inhibition S. agalactiae (19 mm) and K. pneumonia (17 mm). Conclusions The antimicrobial activities of all the four plant extracts are comparable and their potential as alternative in the treatment of infectious by these microorganisms was present in the fish. Susceptibility testing is conducted on isolates using drugs selected on the basis of their importance to human medicine and use in fish production.

731-733

Authors: OPG Nmorsi, C Isaac, BA Ohaneme, HAK Obiazi

Objective To investigate the pro-inflammatory cytokines profiles in in Nigerian pregnant women infected with Plasmodium falciparum (P. falciparum) malaria. Methods Peripheral, and placental blood samples were collected from 96 consenting volunteers comprising 76 P. falciparium infected pregnant women and 20 healthy uninfected pregnant women in Ekpoma, Nigeria, and subjected to ELISA for cytokines evaluation. Results Increased serum concentrations of interferon-gamma (IFN-γ) was observed in infected pregnant women than their uninfected counterparts [(31.2±20.9) pg/mL vs (1.8±0.9) pg/mL] and these differences were statistically significant (χ2 = 26.18, P <0.05). The depressed levels of interleukin-12 (IL-12) seen in peripheral blood of the infected pregnant women than the uninfected women [(13.9±3.6) pg/mL vs (28.4±5.28) pg/mL] respectively was not statistically significant (χ2 = 4.96, P >0.05). The interleukin -6 (IL-6) was significantly elevated in infected pregnant women (81.0±26.1 pg/mL) than in the uninfected pregnant women [(25.0±5.0) pg/mL] (χ2 = 29.58, P <0.05). In all, mean cytokines concentration of IL-6, IL-12 and IFN-γ in the placental blood from infected pregnant women were (53.5±23.4) pg/mL, (8.7±6.9) pg/mL and (16.4±4.0) pg/mL, respectively. The multigravidae had a higher haemoglobin level of 10.2 g/dL and birth weight of 3 000 g than the primigrivadae with lower haemoglobin level of 7.5 g/dL and birth weight of 2 430 g. Conclusions The elevated IFN-γamong the malarous pregnant women implicates it as the major cytokine mediator in the host responses to systematic P. falciparum malaria in our locality.

738-740

Authors: J Meenupriya, M Thangaraj

Objective To isolate and characterize the bioactive secondary metabolite from Callyspongia spp. associated fungi. Methods In vitro antibacterial screening of fungi associated with Callyspongia species, collected from south east coast of India, against selected clinical isolates of bacteria were conducted in this study. The extracts showing good antimicrobial activity were subjected to further analysis to identify the active constituents sponge associated fungi (both biomass and filtrate) with five different solvents. The compound responsible for bioactivity was characterized using Fouvier-transform infrared (FT-IR) and gas chromatography-mass spectrometry (GC-MS) instrumental analysis to identify the functional group and compound. The molecular characterization of the elite fungal strains were done by isolating their genomic DNA and amplify the internal transcribed spacer (ITS) region of 5.8 sr RNA using specific ITS primer. The novelty of the strain was proved by BlastN analysis against non-redundant (NR) database and hence was submitted to GenBank. Results Active compound was Desmethylnomifensine confirmed by GC-MS and the potent fungi was Aspergillus flavus GU815344. Conclusions The isolate exhibits a marked antagonistic activity against potential bacterial pathogens thus illuminating the advanced researches in this decade to focus on clinical pharmacology to identify novel therapeutic targets. The present study depicts a promising scenario to focus on Aspergillus flavus derived compounds which can be easily scaled up for large biomass production and stable formulation as a drug.

745-747

Authors: Sami Bdir, Ghaleb Adwan

Objective To assess the prevalence of intestinal parasites among Jenin Governorate (Northern Palestine) population. Methods A retrospective laboratory analysis of stool specimens was carried out for intestinal parasite examination in Jenin Governmental Hospital, Jenin Governorate, Northern Palestine. The records were collected from the clinical microbiology laboratory of the Jenin Governmental Hospital between January 2000 and December 2009. Results Our retrospective study showed that the prevalence of intestinal parasite infection during 10 years ranged from 32.0-41.5%. There are at least 7 different parasites encountered. The most common pathogenic parasites identified were: Entamoeba histolytica (8.2-18.2%), Enterobius vermicularis (15.6-28.9%). The other parasites present were Giardia lamblia, Ascaris lumbricoides, Strongyloides stercoralis, Taenia species and Ancylostoma duodenale (Hookworms). Conclusions Intestinal parasitic infection is an important public health problem in Jenin governorate, Palestine. It is necessary to develop effective prevention and control strategies including health education and environmental sanitation improvement.

754-756

Authors: Paul O Dienye, Geraldine U Ndukwu

Tuberculous otitis media is a rare disease, hence not often considered in the differential diagnosis of otorrhea. This results in late diagnosis with resulting complications such as irreversible hearing loss. A case report with review of the literature is presented, emphasizing that tuberculosis should be considered in the differential diagnosis of otorrhea not responding to commonly prescribed antibiotics. We also emphasize the importance of awareness creation in the management of such a disease, involving family members to oversee treatment and also research on ways of shortening duration of treatment to avoid default in treatment.