Asian Pacific Journal of Tropical Biomedicine

267-272

Authors: Fouad Yousif, Sherif Hafez, Samia El Bardicy, Menerva Tadros, Hoda Abu Taleb

Objective To test Candonocypris novaezelandiae (Baird) (C. novaezelandiae), sub-class Ostracoda, obtained from the Nile, Egypt for its predatory activity on snail, Biomphalaria alexandrina (B. alexandrina), intermediate host of Schistosoma mansoni (S. mansoni) and on the free-living larval stages of this parasite (miracidia and cercariae). Methods The predatory activity of C. novaezelandiae was determined on B. alexandrina snail (several densities of eggs, newly hatched and juveniles). This activity was also determined on S. mansoni miracidia and cercariae using different volumes of water and different numbers of larvae. C. novaezelandiae was also tested for its effect on infection of snails and on the cercarial production. Results C. novaezelandiae was found to feed on the eggs, newly hatched and juvenile snails, but with significant reduction in the consumption in the presence of other diet like the blue green algae (Nostoc muscorum). This ostracod also showed considerable predatory activity on the free-living larval stages of S. mansoni which was affected by certain environmental factors such as volume of water, density of C. novaezelandiae and number of larvae of the parasite. Conclusions The presence of this ostracod in the aquatic habitat led to significant reduction of snail population, infection rate of snails with schistosme miracidia as well as of cercarial production from the infected snails. This may suggest that introducing C. novaezelandiae into the habitat at schistosome risky sites could suppress the transmission of the disease.

280-283

Authors: Naveed Muhammad, Muhammad Saeed, Haroon Khan, Naila Raziq, Syed Muhammad Ashhad Halimi

Objective To investigate the antipyretic and anticonvulsant activities of n-hexane fraction of Viola betonicifolia (V. betonicifolia). Methods The antipyretic effect was scrutinized using brewer's yeast induced pyrexia and anticonvlsion effect was tested using pentylenetetrazol and strychnine induced convulsion in mice. Results N-hexane fraction of V. betonicifolia demonstrated highly significant antipyretic activity during various assessment times (1-5 h) when challenged in yeast induced pyrexia test. The effect was in a dose dependent manner with maximum attenuation (82.50%) observed at 300 mg/kg i.p. When tested in pentylenetetrazol induced convulsion test, the 1st stage (Ear and facial twitching) and 2nd stage (Convulsive wave through the body) was 100% protected during 24 h at all the test doses (300, 400 and 500 mg/kg i.p.), while the latency time of remaining stages was significantly increased. The maximum effect was observed by n-hexane fraction of V. betonicifolia at 400 and 500 mg/kg i.p., as the latency time for generalized clonic-tonic seizure (5th stage) was increased up to 25.34 min. However, n-hexane fraction of V. betonicifolia had no protection in strychnine induced convulsion test. Conclusions In conclusion, phytopharmacological studies provide scientific foundation to the folk uses of the plant in the treatment of pyrexia and neurological disorders.

291-296

Authors: Ramesh Subramani, Rohitesh Kumar, Pritesh Prasad, William Aalbersberg

Objective To Isolate, purify, characterize, and evaluate the bioactive compounds from the sponge-derived fungus Penicillium sp. FF001 and to elucidate its structure. Methods The fungal strain FF001 with an interesting bioactivity profile was isolated from a marine Fijian sponge Melophlus sp. Based on conidiophores aggregation, conidia development and mycelia morphological characteristics, the isolate FF001 was classically identified as a Penicillium sp. The bioactive compound was identified using various spectral analysis of UV, high resolution electrospray ionization mass spectra, 1H and 13C NMR spectral data. Further minimum inhibitory concentrations (MICs) assay and brine shrimp cytotoxicity assay were also carried out to evaluate the biological properties of the purified compound. Results Bioassay guided fractionation of the EtOAc extract of a static culture of this Penicillium sp. by different chromatographic methods led the isolation of an antibacterial, anticryptococcal and cytotoxic active compound, which was identified as citrinin (1). Further, citrinin (1) is reported for its potent antibacterial activity against methicillin-resistant Staphylococcus aureus (S. aureus), rifampicin-resistant S. aureus, wild type S. aureus and vancomycin-resistant Enterococcus faecium showed MICs of 3.90, 0.97, 1.95 and 7.81 µg/mL, respectively. Further citrinin (1) displayed significant activity against the pathogenic yeast Cryptococcus neoformans (MIC 3.90 µg/mL), and exhibited cytotoxicity against brine shrimp larvae LD50 of 96 µg/mL. Conclusions Citrinin (1) is reported from sponge associated Penicillium sp. from this study and for its strong antibacterial activity against multi-drug resistant human pathogens including cytotoxicity against brine shrimp larvae, which indicated that sponge associated Penicillium spp. are promising sources of natural bioactive metabolites.

303-306

Authors: Emmanuel Swai, Isidory Mwezimpya, Edward Ulicky, Adam Mbise, Winford Moshy

Objective To establish and estimate incidence of contagious bovine pleuropneumonia (CBPP), using abattoir survey as a diagnostic tool in slaughtered cattle in Northern Tanzania. Methods A total of 4 460 cattle were slaughtered in five abattoirs in 3 northern zone regions (Arusha, Kilimanjaro and Tanga) during the period of January to May 2004. They were examined ante-mortem for ‘pneumonia signs’, and ‘characteristic contagious bovine pleuropneumonia (CBPP) lung lesions’. Results Forty-one (0.91%) of the slaughtered cattle, the majority of which were Tanzania short horn zebu, had gross lung lesions suggestive of CBPP. The prevalence of lesions was significantly (P<0.05) higher in Karatu abattoir compared to others. No animal was detected to have lesion in Bomang' ombe abattoir. The most observed pneumonic signs included labored breathing (90%), dry cough (57%) and mucopurulent nasal discharge (47%). The gross characteristic CBPP pathological lesion, frequently encountered was left lung lesion (47%), pinkish lung (71%) and pleural adhesion (98%). Epidemiological reports show that the CBPP reported outbreaks increased from 19 in 2002, 65 in 2003 and 18 in 2004 (January-March). The corresponding number of reported deaths increased from 137 in 2002, 269 in 2003 and 77 in 2004 (January-March). Conclusions It's concluded from this study that CBPP is a problem in spite of the extensive awareness and vaccination campaigns. Nevertheless, a continued surveillance programme including routine checks of all cattle carcasses at the abattoir and subsequent epidemiological investigation of suspected cases are recommended.

311-313

Authors: Bava Amadeo Javier Bava, Domínguez Cecilia, Troncoso Alcides

Objective To communicate the presence of adult females, rabditoid larvae and eggs of Strongyloides stercoralis (S. stercoralis) in the respiratory secretions obtained by tracheal aspirate from a HIV-negative patient who was suffering from polymyositis, and treated with corticoids and amethopterin and assisted by pneumonia. Methods The respiratory secretions submitted to the Parasitology Laboratory of the Muñiz Hospital were made more concentrated by centrifugation (1 500 r/min for 15 seconds). Wet mount microscopy was performed with the pellet. Results It revealed adult females, rabditoid larvae and eggs of S. stercoralis. Further parasitological studies performed after the start of the treatment with ivermectin on fresh fecal samples, gastric lavages and tracheal aspirates showed scanty mobile filariform and rabditoid larvae of the same parasite. Conclusions The presence of adult female S. stercoralis which has never been observed before in the clinical samples submitted to our Laboratory for investigation can be considered as an indirect marker of the severe immunosupression of the patient.

315-324

Authors: Monali Priyadarsini Mishra, Nagen Kumar Debata, Rabindra Nath Padhy

Objective To record surveillance, antibiotic resistance of uropathogens of hospitalized patients over a period of 18 months. Methods Urine samples from wards and cabins were used for isolating urinary tract infection (UTI)-causing bacteria that were cultured on suitable selective media and identified by biochemical tests; and their antibiograms were ascertained by Kirby-Bauer's disc diffusion method, in each 6-month interval of the study period, using 18 antibiotics of five different classes. Results From wards and cabins, 1 245 samples were collected, from which 996 strains of bacteria belonging to 11 species were isolated, during April 2011 to September 2012. Two Gram-positive, Staphylococcus aureus (S. aureus) and Enterococcus faecalis (E. faecalis), and nine Gram-negative bacteria, Acinetobacter baumannii, Citrobacter sp., Escherichia coli, Enterobacter aerogenes, Klebsiella pneumoniae, Klebsiella oxytoca, Proteus mirabilis, Proteus vulgaris and Pseudomonas aeruginosa were isolated. Both S. aureus and E. faecalis were vancomycin resistant, and resistant-strains of all pathogens increased in each 6-month period of study. Particularly, all Gram-negatives were resistant to nitrofurantoin and co-trimoxazole, the most preferred antibiotics of empiric therapy for UTI. Conclusions Antibiograms of 11 UTI-causing bacteria recorded in this study indicated moderately higher numbers of strains resistant to each antibiotic studied, generating the fear of precipitating fervent episodes in public health particularly with bacteria, Acinetobacter baumannii, Escherichia coli, Klebsiella pneumoniae and S. aureus. Moreover, vancomycin resistance in strains of S. aureus and E. faecalis is a matter of concern.

330-333

Authors: Viroj Wiwanitkit

In the present day, bioinformatics becomes the modern science with several advantages. Several new “omics” sciences have been introduced for a few years and those sciences can be applied in biomedical work. Here, the author will summarize and discuss on important applications of omics studies in microbiology focusing on microbial pathogeny. It can be seen that genomics and proteinomics can be well used in this area of biomedical studies.

174-181

Authors: Saber Sakr, Laila Rashed, Waheba Zarouk, Rania El-Shamy

Objective To evaluate the effect of mesenchymal stem cells (MSCs) in rats with anti-Thy1,1 nephritis. Methods Female albino rats were divided into three groups, control group, anti-Thy1,1 group and treatment with i.v. MSCs group. MSCs were derived from bone marrow of male albino rats, Y-chromosome gene was detected by polymerase chain reaction in the kidney. Serum urea and creatinine were estimated for all groups. Kidney of all studied groups was examined histologically and histochemically (total carbohydrates and total proteins). DNA fragmentation and expression of α-SMA were detected. Results Kidney of animals injected with anti-Thy1,1 showed inflammatory leucocytic infiltration, hypertrophied glomeruli, tubular necrosis and congestion in the renal blood vessels. The kidney tissue also showed reduction of carbohydrates and total proteins together with increase in apoptosis and in expression of α-SMA. Moreover, the levels of urea and creatinine were elevated. Treating animals with MSCs revealed that kidney tissue displayed an improvement in the histological and histochemical changes. Apoptosis and α-SMA expression were decreased, and the levels of urea and creatinine decreased. Conclusions The obtained results demonstrated the potential of MSCs to ameliorate the structure and function of the kidney in rats with anti-Thy1,1 nephritis possibly through the release of paracrine growth factor(s).

186-190

Authors: Alessio Arbuatti, Leonardo Della Salda, Mariarita Romanucci

Objective To describe the occurrence of various spinal deformations in a captive-bred wild line of Poecilia wingei (P. wingei). Methods Fish belonging to a wild line of P. wingei caught from Laguna de Los Patos, Venezuela, were bred in an aquarium home-breeding system during a period of three years (2006-2009). The spinal curvature was observed to study spinal deformities in P. wingei. Results Out of a total of 600 fish, 22 showed different types of deformities (scoliosis, lordosis, kyphosis), with a higher incidence in females. Growth, swimming and breeding of deformed fish were generally normal. Conclusions Possible causes for spinal curvature in fish are discussed on the basis of the current literature. While it is not possible to determine the exact cause(s) of spinal deformities observed in the present study, traumatic injuries, nutritional imbalances, genetic defects or a combination of these factors can be supposed to be involved in the pathogenesis of such lesions.

196-201

Authors: Sevil Albayrak, Ahmet Aksoy

Objective To evaluate total phenolic, antioxidant and antimicrobial activities of methanolic extracts of the Wiedemannia multifida (W. multifida) and Wiedemannnia orientalis (W. orientalis). Methods Phosmomolybdenum assay, DPPH radical scavenging activity and β-carotene-linoleate bleaching were used to evaluate the antioxidant capacity. Results The total phenolics were found to be (22.45±0.60) and (9.53±0.00) mg gallic acid equivalents (GAE)/g in W. multifida and W. orientalis extracts, respectively. The predominant phenolic compounds identified by HPLC-DAD in the both extracts were rutin+ellagic acid and kaempferol. Total antioxidant capacity and DPPH radical scavenging activity of W. multifida were higher than W. orientalis. In β-carotene-linoleic acid system, both extracts exhibited strong inhibition against linoleic acid oxidation. Antimicrobial activity was assessed by the agar diffusion method against fifteen microorganisms. Both extracts exhibited remarkable antibacterial activity. Conclusions The present study suggests that methanolic extracts of W. multifida and W. orientalis could be a good source of antioxidant and antimicrobial agents in foods, pharmaceuticals preparations.

207-210

Authors: Tanasak Changbunjong, Thekhawet Weluwanarak, Namaoy Taowan, Parut Suksai, Tatiyanuch Chamsai, Poonyapat Sedwisai

Objective To investigate the abundance and seasonal dynamics of mosquitoes, and to detect Japanese encephalitis virus (JEV) in these mosquitoes at the nesting colony of ardeid birds. Methods Mosquitoes were collected bimonthly from July 2009 to May 2010 by Centers for Disease Control. Light traps and dry ice, as a source of CO2, were employed to attract mosquitoes. Mosquitoes were first identified, pooled into groups of upto 50 mosquitoes by species, and tested for JEV infection by viral isolation and reverse transcriptase polymerase chain reaction. Results A total of 20 370 mosquitoes comprising 14 species in five genera were collected. The five most abundant mosquito species collected were Culex tritaeniorhynchus (95.46%), Culex vishnui (2.68%), Culex gelidus (0.72%), Anopheles peditaeniatus (0.58%) and Culex quinquefasciatus (0.22%). Mosquito peak densities were observed in July. All of 416 mosquito pools were negative for JEV. Conclusions This study provides new information about mosquito species and status of JEV infection in mosquitoes in Thailand. Further study should be done to continue a close survey for the presence of this virus in the ardeid birds.

217-222

Authors: Hong-Zhu Lu, Jian-Hua Zhou

Objective To investigate the effects of hepatitis B virus (HBV) X protein (HBx) on the expression of tumor necrosis factor-α (TNF-α) in glomerular mesangial cells (GMCs) and the underlying intracellular signal pathways. Methods The plasmid pCI-neo-X that carries the X gene of hepatitis B virus was transfected into cultured GMCs. HBx expression in the transfected GMCs was assessed by Western-blot. TNF-α protein and mRNA were assessed by ELISA and semi-quantitative RT-PCR, respectively. Three kinase inhibitors-U0126, an inhibitor of extracellular signal-regulated kinases (ERKs); lactacystin, an inhibitor of nuclear factor-κB (NF-κB); and SB203580, a selective inhibitor of p38 MAP kinase (p38 MAPK) were used to determine which intracellular signal pathways may underlie the action of HBx on TNF-α expression in transfected GMCs. Results A significant increase in HBx expression in pCI-neo-X transfected GMCs was detected at 36 h and 48 h, which was not affected by any of those kinase inhibitors mentioned above. A similar increase in the expression of both TNF-α protein and mRNA was also observed at 36 h and 48 h, which was significantly decreased in the presence of U0126 or lactacytin, but not SB203580. Conclusions HBx upregulates TNF-α expression in cultured GMCs, possibly through ERKs and NF-κB pathway, but not p38 MAPK pathway.

224-228

Authors: Xin Ee Tan, Hui-min Neoh, Salasawati Hussin, Noraziah Mohamad Zin

Objective To genotypically characterize methicillin-resistant Staphylococcus aureus (MRSA) strains isolated from medical and surgical wards in Universiti Kebangsaan Malaysia Medical Centre (UKMMC) in 2009. Methods MRSA strains were collected and molecularly typed by pulsed-field gel electrophoresis (PFGE). Results PFGE typing on 180 MRSA isolated in UKMMC identified 5 pulsotypes (A-E) and 6 singletons, where pulsotypes B and C were suspected to be divergent clones originating from a single ancestor. This study also showed that most MRSA strains were isolated from swab (119 isolates), followed by blood (22 isolates), tracheal aspirate (11 isolates) and sputum (10 isolates). On the other hand, urine and bone isolates were less, which were 4 and 1 isolates, respectively. The distribution of different pulsotypes of MRSA among wards suggested that MRSA was communicated in surgical and medical wards in UKMMC, with pulsotype B MRSA as the dominant strain. Besides, it was found that most deceased patients were infected by pulsotype B MRSA, however, no particular pulsotype could be associated with patient age, underlying disease, or ward of admittance. Conclusions Five pulsotypes of MRSA and 6 singletons were identified, with pulsotype B MRSA as the endemic strains circulating in these wards, which is useful in establishment of preventive measures against MRSA transmission.

232-236

Authors: Sobia Idrees, Usman A Ashfaq

Dengue virus infection has become a global threat affecting around 100 countries in the world. Currently, there is no licensed antiviral agent available against dengue. Thus, there is a strong need to develop therapeutic strategies that can tackle this life threatening disease. RNA interference is an important and effective gene silencing process which degrades targeted RNA by a sequence specific process. Several studies have been conducted during the last decade to evaluate the efficiency of siRNA in inhibiting dengue virus replication. This review summarizes siRNAs as a therapeutic approach against dengue virus serotypes and concludes that siRNAs against virus and host genes can be next generation treatment of dengue virus infection.

244-252

Authors: Anwesa Bag, Subir Kumar Bhattacharyya, Rabi Ranjan Chattopadhyay

Medicinal plants are part and parcel of human society to combat diseases from the dawn of civilization. Terminalia chebula Retz. (Fam. Combretaceae), is called the ‘King of Medicine’ in Tibet and is always listed at the top of the list of ‘Ayurvedic Materia Medica’ because of its extraordinary power of healing. The whole plant possesses high medicinal value and traditionally used for the treatment of various ailments for human beings. Some of the folklore people used this plant in the treatment of asthma, sore throat, vomiting, hiccough, diarrhea, dysentery, bleeding piles, ulcers, gout, heart and bladder diseases. The plant has been demonstrated to possess multiple pharmacological and medicinal activities, such as antioxidant, antimicrobial, antidiabetic, hepatoprotective, anti-inflammatory, antimutagenic, antiproliferative, radioprotective, cardioprotective, antiarthritic, anticaries, gastrointestinal motility and wound healing activity. But no systematic updated information on the therapeutic effectiveness of Terminalia chebula, a popular herbal remedy in India and South-East Asia has so far been reported. This review highlights an updated information particularly on the phytochemistry and various pharmacological and medicinal properties of Terminalia chebula Retz. and some of its isolated compounds, along with their safety evaluation. This may provide incentive for proper evaluation of the plant as medicinal agent against the human diseases and also to bridge the lacunae in the existing literature and future scope which may offer immense opportunity for researchers engaged in validation of the traditional claims and development of safe and effective botanical medicine.

89-94

Authors: Freddie Masaninga, Emmanuel Chanda, Pascalina Chanda-Kapata, Busiku Hamainza, Hieronymo T Masendu, Mulakwa Kamuliwo, Wambinji Kapelwa, John Chimumbwa, John Govere, Mac Otten, Ibrahima Soce Fall, Olusegun Babaniyi

A comprehensive desk review of malaria trends was conducted between 2000-2010 in Zambia to study malaria epidemiology and trends to guide strategies and approaches for effective malaria control. This review considered data from the National Health Information Management System, Malaria Surveys and Programme Review reports and analyzed malaria in-patient cases and deaths in relation to intervention coverage for all ages. Data showed three distinct epidemiological strata after a notable malaria reduction (66%) in in-patient cases and deaths, particularly between 2000-2008. These changes occurred following the (re-)introduction and expansion of indoor residual spraying up to 90% coverage, scale-up of coverage of long-lasting insecticidetreated nets in household from 50% to 70%, and artemisin-based combination therapy nationwide. However, malaria cases and deaths re-surged, increasing in 2009-2010 in the northern-eastern parts of Zambia. Delays in the disbursement of funds affected the implementation of interventions, which resulted in resurgence of cases and deaths. In spite of a decline in malaria disease burden over the past decade in Zambia, a reversal in impact is notable in the year 2009-2010, signifying that control gains are fragile and must be sustained toeliminate malaria.

100-104

Authors: Wabo Poné J, Payne V K, Mbogning Tayo Gertrude, Komtangi Marie Claire, Yondo Jeannette, Ngangout Alidou M, Mpoame Mbida, Bilong Bilong CF

Objective: To evaluate the ovicidal and larvicidal activities of aqueous and ethanolic extracts of leaves of Dichrocephala integrifolia (D. integrifolia) against the eggs (fresh and embryonnated), the first and second larval stages of Heligmosomoides bakeri. In order to verify if this medicinal plant possesses active compounds capable of inhibiting the embryonation and hatching of eggs or to induce the mortality of larvae (L1 and L2). Methods: Dried extracts were diluted in distilled water to obtain five different concentrations: 625, 1250, 2500, 3750 and 5000 μg/mL. Fresh eggs obtained from artificially infected mice feces were exposed to these different concentrations for 48 h. Time of contact for embryonated eggs was 6 h while L1 and L2 larvae were exposed for 24 h. Distilled water (placebo) and 1.5% DMSO were used as negative controls. Results: Distilled water, and 1.5% DMSO had no effect on embryonation, hatching and larval survival. Aqueous extracts of D. integrifolia showed a weak activity against all stages of the parasite at all concentrations tested. On the contrary, the ethanolic extract of D. integrifolia inhibited the embryonation of 87.5% of fresh eggs, the hatching of 81.1% of embryonated eggs and induced the mortality of 98.1% and 98% of L1 and L2 larvae respectively at 5 000 μg/mL. Conclusions: The results of the present study indicate that the ethanolic extracts of D. integrifolia contained compounds with ovicidal and larvicidal properties. In spite of these results, in vivo tests, studies on toxicity and mechanism of action of active compounds are also needed to validate the utilisation of this medicinal plant by population of Dschang-Cameroon to treat gastro-intestinal parasites.

111-117

Authors: Zulfaidah Penata Gama, Nobukazu Nakagoshi, Suharjono, Faridah Setyowati

Objective To investigate the toxicity of indigenous Bacillus thuringiensis (B. thuringiensis)isolates from Malang City for controlling Aedes aegypti (Ae. aegypti) larvae. Methods Soil samples were taken from Purwantoro and Sawojajar sub-districts. Bacterial isolation was performed using B. thuringiensis selective media. Phenotypic characteristics of the isolates were obtained with the simple matching method. The growth and prevalence of spores were determined by the Total Plate Count method, and toxicity tests were also performed on the third instar larval stage of Ae. aegypti. The percentage of larval mortality was analysed using probit regression. The LC50 was analysed by ANOVA, and the Tukey HSD interval was 95%. Results Among the 33 selected bacterial isolates, six were obtained (PWR4-31, PWR4-32, SWJ4-2b, SWJ4-4b, SWJ-4k and SWJ5-1) that had a similar phenotype to reference B. thuringiensis. Based on the dendrogram, all of the bacterial isolates were 71% similar. Three isolates that had a higher prevalence of reference B. thuringiensis were PWR4-32, SWJ4-4b and SW5-1, of which the spore prevalence was 52.44%, 23.59%, 34.46%, respectively. These three indigenous isolates from Malang City successfully killed Ae. aegypti larvae. The PWR4-32 isolates were the most effective at killing the larvae. Conclusions Six indigenous B. thuringiensis isolates among the 33 bacterial isolates found in the Sawojajar and Purwantoro sub-districts were toxic to the third instar larvae of Ae. aegypti. The PWR4-32 isolates were identical to the reference B. thuringiensis and had 88% phenotype similarity. The PWR4-32 isolates had the highest spore prevalence (52.44%), and the early stationary phase occurred at 36 h. The PWR4-32 isolates were the most effective at killing Ae. aegypti larvae (LC50-72 h=2.3×108 cells/mL).

123-128

Authors: Abdollah Ghasemi Pirbalouti, Maryam Mohammadi

Objective To examine the chemical variability in inflorescences of wild populations of Stachys lavandulifolia Vahl (S. lavandulifolia) collected throughout two provinces (Isfahan and Chaharmahal va Bakhtiary), Southwest Iran. Methods The essential oils of S. lavandulifolia Vahl from seven locations were obtained by hydro-distillation and analysed by gas chromatography and gas chromatography-mass spectrometry. Results The results revealed that distinct differences in the content of compounds depending on region of sample collection. The main constituents of the essential oils were α-thujone (0.3%-32.3%), α-pinene (trace to 37.3%), myrcene (0.5%-15.9%), β-phellandrene (1.1%-37.9%), germacrene D (0.4%-11.3%), Δ-cadinene (trace to 11.6%) and 1, 4-methano-1 H-indene (trace to 10.1%). Conclusions The results of the present study indicated that essential oil components of S. lavandulifolia Vahl can be varied with genetic (ecotype), environmental conditions and geographic origin. In general, the essential oils of various populations of S. lavandulifolia Vahl were rich in monoterpenoids and sesquiterpenoids.

140-145

Authors: Reza Mirnejad, Sepideh Mostofi, Faramaz Masjedian

Objective To investigate antibiotic resistance and carriage class 1 and 2 integrons in clinical isolates of Acinetobacter baumannii (A. baumannii) from Tehran, Iran. Methods Antimicrobial susceptibility testing was performed according to the Clinical and Laboratory Standards Institute. The presence of integrons was investigated by PCR using specific primers. Results Among isolated A. baumannii strains, 82% were multidrug resistant, 27 samples (54%) were resistant to three or more than three antibiotics and 16 samples (32%) showed resistance to two antibiotics. Integrons were detected from 44 of 50 isolates (88%), with classes 1 and 2 being observed in 42% (21/50) and 82% (41/50) of isolates, respectively. Integron-positive A. baumannii isolates showed higher antibiotic resistance than integron-negative isolates and all showed a multidrug-resistant phenotype. Conclusions Our findings show that classes 1 and 2 integrons, and especially classes 2 integrons are widely disseminated among A. baumannii strains isolated from Tehran and these structures are playing a major role in the acquisition of multidrug resistance in these strains. So monitoring of drug resistance with investigating carriage class 1 and 2 integrons is very important to plan specific infection control measures due to multidrug resistance A. baumannii in Iran hospitals.

148-153

Authors: Debasmita Dubey, Shakti Rath, Mahesh C. Sahu, Subhrajita Rout, Nagen K. Debata, Rabindra N. Padhy

Objective To investigate the infection of hospital- and community-acquired “erythromycin-induced clindamycin resistant” strains or D-test positives of clinical isolates of Staphylococcus aureus (S. aureus) (with and without methicillin resistance) in a hospital. Methods Strains of S. aureus isolated from clinical specimens were subjected to D-test and antibiotic profiling. Results Of the total 278 isolates, 140 (50.35%) were D-test positives and the rest were D-test negatives. Further, of 140 (100%) positives, 87 (62.14%) and 53 (37.85%) strains were from males and females, respectively. Of 140 (100%) positives, 117 (83.57%) were methicillin resistant S. aureus and 23 (16.42%) were methicillin sensitive S. aureus; of 140 strains, 103 (73.57%) strains from persons with and 37 (26.42%) were without related infections; of 140 strains, 91 (65%) and 49 (35%) were from hospital- and community-acquired samples, respectively. In 140 strains, 118 (84.28%) with comorbidities and 22 (15.71%) without comorbidities cases were recorded; similarly, persons with prior antibiotic uses contributed 108 (77.14%) and without 32 (22.85%) positive strains. These binary data of surveillance were analyzed by a univariate analysis. It was evident that the prior antibiotic uses and comorbidities due to other ailments were the determinative factors in D-test positivity, corroborated by low P values, P=0.001 1 and 0.002 4, respectively. All isolates (278) were resistant to 17 antibiotics of nine groups, in varying degrees; the minimum of 28% resistance for vancomycin and the maximum of 97% resistance for gentamicin were recorded. Further, of 278 strains, only 42 (15.1%) strains were resistant constitutively to both antibiotics, erythromycin resistant and clindamycin resistant, while 45 (16.2%) strains were constitutively sensitive to both antibiotics (erythromycin sensitive and clindamycin sensitive). Further, of the rest 191 (68.7%) strains were with erythromycin resistant and clindamycin resistant, of which only 140 (50.35%) strains were D-test positives, while the rest 51 (18.34%) strains were D-test negatives. Conclusions In view of high prevalence of D-test positive S. aureus strains, and equally high prevalence of multidrug resistant strains both in community and hospital sectors, undertaking of D-test may be routinely conducted for suppurative infections.

156-162

Authors: Vivek Kumar Vyas, Keyur Brahmbhatt, Hardik Bhatt, Utsav Parmar

Many active secretions produced by animals have been employed in the development of new drugs to treat diseases such as hypertension and cancer. Snake venom toxins contributed significantly to the treatment of many medical conditions. There are many published studies describing and elucidating the anti-cancer potential of snake venom. Cancer therapy is one of the main areas for the use of protein peptides and enzymes originating from animals of different species. Some of these proteins or peptides and enzymes from snake venom when isolated and evaluated may bind specifically to cancer cell membranes, affecting the migration and proliferation of these cells. Some of substances found in the snake venom present a great potential as anti-tumor agent. In this review, we presented the main results of recent years of research involving the active compounds of snake venom that have anticancer activity.

8-16

Authors: Kajal Chakraborty, Nammunayathuputhenkotta Krishnankartha Praveen, Kodayan Kizekadath Vijayan, Gonugontla Syda Rao

Objective: To evaluate the antioxidant activities and total phenolic contents of brown seaweeds belonging to Turbinaria spp. [Turbinaria conoides (T. conoides) and Turbinaria ornata (T. ornata)collected from Gulf of Mannar of southeastern coast of India in various in vitro systems. Methods:The antioxidant activity was evaluated using different in vitro systems, viz., 1,1-diphenyl-2-picrylhydrazyl (DPPH), 2, 2′-azino-bis-3ethylbenzothiozoline-6-sulfonic acid diammonium salt(ABTS), H2O2/HO radical scavenging, Fe2+ ion chelating ability, and reducing potential. FolinCiocalteu method was used to determine the total phenolic content of the extracts, and the results were expressed as mg of gallic acid equivalents (GE)/g of the seaweed extracts. Thiobarbituric acid-reactive substances assay was employed to assess the ability of the seaweed extracts to inhibit lipid oxidation. Results: Ethyl acetate (EtOAc) fraction of T. conoides registered significantly higher phenolic content (105.97 mg GE/g) than that of T. ornata (69.63 mg GE/g). Significantly higher antioxidant potential as determined by DPPH (64.14%) radical scavenging acti vity was registered in EtOAc fraction of T. ornata. A higher ABTS.+ radical scavenging (IC50 3.16 µg/mL),Fe2+ chelating (IC50 0.46 mg/mL), H2O2 scavenging (IC50 4.25 mg/mL), lipid peroxidation inhibitory (TBARS, IC50 0.21 mg/mL), and reducing abilities (IC50 52.67 mg/mL) (P<0.05) were realized in EtOAc fraction of T. ornata than other fractions. Conclusions: This study indicated the potential use of T. conoides and T. ornata as candidate species to be used as food supplements/functional foods to increase shelf-life of food items for human consumption, and nutraceuticals to deter deleterious free radical-induced life-threatening diseases.

22-27

Authors: Shahrokh Bagheri, Hassan Ahmadvand, Ali Khosrowbeygi, Farshid Ghazanfari, Narges Jafari, Habibolah Nazem, Reza Haji Hosseini

Objective: To assess various antioxidative activities of Satureja khozestanica essential oil(SKE) and its effect on oxidation of low density lipoprotein (LDL) induced by CuSO4 in vitro by monitoring the formation of conjugated dienes and malondialdehyde (MDA). Methods: The formation of conjugated dienes, lag time and MDA were measured. Inhibition of this Cu-induced oxidation was studied in the presence of several concentrations of SKE. Also total antioxidant activity and free radical scavenging of SKE were determinated. Results: It was demonstrated that SKE was able to inhibit LDL oxidation and decrease the resistance of LDL against oxidation. The inhibitory effects of SKE on LDL oxidation were dose-dependent at concentrations ranging from 50 to 200 μg/mL. Total antioxidant capacity of SKE was (3.20±0.40) nmol of ascorbicacid equivalents/g SKE. The SKE showed remarkable scavenging activity on 2, 2-diphenylpicrylhydrazyl, IC50 (5.30± 0.11) ng/mL. Conclusions: This study shows that SKE is a source of potent antioxidants and prevents the oxidation of LDL in vitro and it may be suitable for use in food and pharmaceutical applications.

35-40

Authors: Nabila Benariba, Rabeh Djaziri, Wafaa Bellakhdar, Nacera Belkacem, Marcel Kadiata, Willy J. Malaisse, Abdullah Sener

Objective: To study the phytochemical screening of different extracts from Citrullus colocynthis (C. colocynthis ) seeds extracts and to assess their antioxidant activity on the DPPH free radical scavenging. Methods: Phytochemical screening, total content of polyphenols and flavonoids of C. colocynthis seeds extracts, including a crude aqueous extract (E1), a defatted aqueous extract (E2), a hydromethanolic extract (HM), an ethyl acetate extract (EA) and a n-butanol extract (n-B) was carried out according to the standard methods and to assess their corresponding effecton the antioxidant activity of this plant. Results: None of these extracts contained detectable amount of alkaloid, quinone, antraquinone, or reducing sugar. Catechic tannins and flavonoids were abundant in E1, HM and EA, whilst terpenoids were abundantly present in E1 and n-B but only weekly in HM. Coumarins were found in E2, EA and n-B. Polyphenols, expressed as gallic acid equivalent, amounted, per 100 g plant matter, to 329, 1002 and 150 mg in EA, HM an E1 respectively. Flavonoids, expressed as catechin equivalent, amounted, per 100 g plant matter to 620, 241 and 94 mg in EA, HM and E1 respectively. Comparable values were found in n-B and E1, with lower values in E2. Quercetin, myricetin and gallic acid were found in the EA and HM extracts by thin layer chromatography, The antioxidative effect of these extracts yielded, when tested at a concentration of 2 000 μg/mL in a 1,1-diphenyl-2-picrylhydrazyl assay, a reducing percentage of 88.8% with EA, 74.5% with HM and 66.2% with E1, and corresponding IC50 of 350, 580 and 500 μg/mL as compared to 1.1 μg/mL for ascorbic acid. Conclusions: These qualitative and quantitative analytical data document the presence in C. colocynthis extracts of such chemical compounds as flavonoids responsible for the antioxidant activity, as well as other biological activities of this plant.

47-52

Authors: Sirikamon Saengmee-anupharb, Toemsak Srikhirin, Boonyanit Thaweboon, Sroisiri Thaweboon,Taweechai Amornsakchai, Surachai Dechkunakorn, Theeralaksna Suddhasthira

Objective: To evaluate the antimicrobial activities of silver inorganic materials, including silver zeolite (AgZ), silver zirconium phosphate silicate (AgZrPSi) and silver zirconium phosphate (AgZrP), against oral microorganisms. In line with this objective, the morphology and structure of each type of silver based powders were also investigated. Methods: The antimicrobial activities of AgZ, AgZrPSi and AgZrP were tested against Streptococcus mutans, Lactobacillus casei, Candida albicans and Staphylococcus aureus using disk diffusion assay as a screening test. The minimum inhibitory concentration (MIC) and minimum lethal concentration (MLC) were determined using the modified membrane method. Scanning electron microscope and X-ray diffraction were used to investigate the morphology and structure of these silver materials. Results: All forms of silver inorganic materials could inhibit the growth of all test microorganisms. The MIC of AgZ, AgZrPSi and AgZrP was 10.0 g/L whereas MLC ranged between 10.0-60.0 g/L. In terms of morphology and structure, AgZrPSi and AgZrP had smaller sized particles (1.5-3.0 µm) and more uniformly shaped than AgZ. Conclusions: Silver inorganic materials in the form of AgZ, AgZrPSi and AgZrP had antimicrobial effects against all test oral microorganisms and those activities may be influenced by the crystal structure of carriers. These results suggest that these silver materials may be useful metals applied to oral hygiene products to provide antimicrobial activity against oral infection.

58-63

Authors: Ghassan Mohammad Sulaiman, Wasnaa Hatif Mohammed, Thorria Radam Marzoog, Ahmed Abdul Amir AlAmiery, Abdul Amir H. Kadhum, Abu Bakar Mohamad

Objective: To synthesize silver nanopaticles from leaves extract of Eucalyptus chapmaniana (E. chapmaniana) and test the antimicrobial of the nanoparticles against different pathogenic bacteria, yeast and its toxicity against human acute promyelocytic leukemia (HL-60) cell line. Methods: Ten milliliter of leaves extract was mixed with 90 mL of 0.01 mmol/mL or 0.02 mmol/mL aqueous AgNO3 and exposed to sun light for 1 h. A change from yellowish to reddish brown color was observed. Characterization using UV-vis spectrophotometery and X-ray diffraction analysis were performed. Antimicrobial activity against six microorganisms was tested using well diffusion method and cytoxicity test using 3-(4, 5-Dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide, a yellow tetrazole was obtained on the human leukemia cell line (HL-60). Results: UVvis spectral analysis showed silver surface plasmon resonance band at 413 nm. X-ray diffraction showed that the particles were crystalline in nature with face centered cubic structure of the bulk silver with broad beaks at 38.50 ° and 44.76 °. The synthesized silver nanoparticles efficiently inhibited various pathogenic organisms and reduced viability of the HL-60 cells in a dosedependent manner. Conclusions: It has been demonstrated that the extract of E. chapmaniana leaves are capable of producing silver nanoparticles extracellularly and the Ag nanoparticles are quite stable in solution. Further studies are needed to fully characterize the toxicity and the mechanisms involved with the antimicrobial and anticancer activity of these particles.

69-73

Authors: Laila Salim Al Hashmi, Mohammad Amzad Hossain, Afaf Mohammed Weli, Qasim Al-Riyami, Jamal Nasser AlSabahi

Objective: To isolate and analyze the chemical composition in different crude extracts of from the leaves of locally grown of Thymus vulgaris L (T. vulgaris) by gas chromatographymass spectrometry (GC-MS). Methods: The shade dried leaves powder was extracted with methanol by using Soxhlet extractor. Methanol crude extracts of T. vulgaris and the derived fractions of hexane, chloroform, ethyl acetate and butanol were obtained. Results: Qualitative analyses of various organic crude extracts of T. vulgaris by using GC-MS showed that there were different types of high and low molecular weight compounds. Most of the isolated and identified compounds by GC-MS in the crude extracts are basically biologically important. Further, the T.vulgaris leaf possessed certain characteristics that can be ascribed to cultivation on a domestic plantation. The crude extracts were prepared from the powder leaves of T. vulgaris for respective compounds can be chosen on the basis of above GC-MS analysis. Conclusions: All the major compounds were identified and characterized by spectroscopic method in different organic crude extracts of T. vulgaris are biologically active molecules. Thus the identification of a good number of compounds in various crude extracts of T. vulgaris might have some ecological role.

79-84

Authors: Rajamani Saranya, Thirunavukkarasu Thirumalai, Munisami Hemalatha, Ranganathan Balaji, Ernest David

Traditional medicine system of India comprised varieties of plants which are playing a significant role in curing diseases from ancient times. Among them, Enicostemma littorale blume (E.littorale) a perennial herb of the family Gentianaceae is cosmopolitan in occurrence in India. The bittery natured plant acts as a laxative, helps in curing fever, rheumatism, skin diseases, abdominal disorders, snake bite, obesity and helps to regulate blood sugar levels. The plant constituents have been reported for possessing antimicrobial, antioxidant, antiulcer, antiinflammatory, hypolipidaemic, hepatoprotective and hypoglycemic properties. This review provides a bird’s eye view about geographical distribution, physicochemical parameters, phytoconstituents and pharmacological properties of E. littorale.

930–935

Authors: P Kumar, S Senthamil Selvi, M Govindaraju

Objective To investigate the anti-biofilm and anti-bacterial activity of Junceella juncea (J. juncea) against biofilm forming pathogenic strains. Methods Gorgonians were extracted with methanol and analysed with fourier transform infrared spectroscopy. Biofilm forming pathogens were identified by Congo red agar supplemented with sucrose. A quantitative spectrophotometric method was used to monitor in vitro biofilm reduction by microtitre plate assay. Anti-bacterial activity of methanolic gorgonian extract (MGE) was carried out by disc diffusion method followed by calculating the percentage of increase with crude methanol (CM). Results The presence of active functional group was exemplified by FT-IR spectroscopy. Dry, black, crystalline colonies confirm the production of extracellular polymeric substances responsible for biofilm formation in Congo red agar. MGE exhibited potential anti-biofilm activity against all tested bacterial strains. The anti-bacterial activity of methanolic extract was comparably higher in Salmonella typhii followed by Escherichia coli, Vibrio cholerae and Shigella flexneri. The overall percentage of increase was higher by 50.2% to CM. Conclusions To conclude, anti-biofilm and anti-bacterial efficacy of J. juncea is impressive over biofilm producing pathogens and are good source for novel anti-bacterial compounds.

944–947

Authors: Rabia Nazm, Asghari Bano

Objective To investigate the in vitro antimicrobial activities of the leaf extract in different solvents viz., methanol, ethanol and water extracts of the selected plant Ricinus communis. Methods Agar well diffusion method and agar tube dilution method were carried out to perform the antibacterial and antifungal activity of methanol, ethanol and aqueous extracts. Results Methanol leaf extracts were found to be more active against Gram positive bacteria (Bacillus subtilis: ATCC 6059 and Staphylococcus aureus: ATCC 6538) as well as Gram negative bacteria (Pseudomonas aeruginosa: ATCC 7221 and Klebsiella pneumoniae) than ethanol and aqueous leaf extracts. Antifungal activity of methanol and aqueous leaf extracts were also carried out against selected fungal strains as Aspergillus fumigatus and Aspergillus flavus. Methanolic as well as aqueous leaf extracts of Ricinus communis were effective in inhibiting the fungal growth. Conclusions The efficient antibacterial and antifungal activity of Ricinus communis from the present investigation revealed that the methanol leaf extracts of the selected plant have significant potential to inhibit the growth of pathogenic bacterial and fungal strains than ethanol and aqueous leaf extracts.

953–959

Authors: Swetha Sunkar, C Valli Nachiyar

Objective To synthesize the ecofriendly nanoparticles, which is viewed as an alternative to the chemical method which initiated the use of microbes like bacteria and fungi in their synthesis. Methods The current study uses the endophytic bacterium Bacillus cereus isolated from the Garcinia xanthochymus to synthesize the silver nanoparticles (AgNPs). The AgNPs were synthesized by reduction of silver nitrate solution by the endophytic bacterium after incubation for 3-5 d at room temperature. The synthesis was initially observed by colour change from pale white to brown which was confirmed by UV-Vis spectroscopy. The AgNPs were further characterized using FTIR, SEM-EDX and TEM analyses. Results The synthesized nanoparticles were found to be spherical with the size in the range of 20-40 nm which showed a slight aggregation. The energy-dispersive spectra of the nanoparticle dispersion confirmed the presence of elemental silver. The AgNPs were found to have antibacterial activity against a few pathogenic bacteria like Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, Salmonella typhi and Klebsiella pneumoniae. Conclusions The endophytic bacteria identified as Bacillus cereus was able to synthesize silver nanoparticles with potential antibacterial activity.

966–974

Authors: Sivagnanam Selva Kumar, Mudiganti Ram Krishna Rao, Maruthaiveeran Periyasamy Balasubramanian

Objective To find out the anticancer effect of Indigofera aspalathoides (I. aspalathoides) on 20-methylcholanthrene induced fibrosarcoma in rats. Methods Fibrosarcoma was induced in Wistar strain male albino rats by 20-methylcholanthrene. Intraperitoneous (i.p.) administration of 250 mg/kg body weight/day of aqueous extract of I. aspalathoides for 30 d effectively suppressed chemically induced tumors. Parameters such as body weight, liver and kidney weight, tumor weight, mean survival time, behavioral changes, blood glucose, blood glycogen and marker enzymes such as alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), acid phosphatase (ACP) and 5′-nucleiotidase (5′-NT) in serum, liver and kidney and lipid profiles such as total cholesterol, phospholipids, free fatty acids in liver and kidney of control and experimental animals were studied. Results Fibrosarcoma bearing animals were ferocious and anxious. The mean survival time was found to increase after the treatment. The body weights were significantly decreased (P<0.001) in group II fibrosarcoma animals which steadily increased after the treatment with I. aspalathoides. The liver and kidney weights were significantly increased whereas the tumor weights decreased as compared to the weights in untreated fibrosarcoma bearing rats. The blood glucose and the liver and kidney glycogen levels were found to decrease significantly (P<0.001) in group II animals. Elevated activities of marker enzymes were observed in serum, liver and kidney of fibrosarcoma bearing Group II animals which were normalize after I. aspalathoides treatment. In the liver and kidney of Group II animals the total cholesterol increased whereas the phospholipids and free fatty acid levels decreased (P<0.001) which were normalized after treatment. Conclusions The treatment by I. aspalathoides on fibrosarcoma bearing rats has improved the levels of various parameters indicating its antiproliferative and anticancer activity.

981–986

Authors: Alagarmalai Jeyasankar, Selvaraj Premalatha, Kuppusamy Elumalai

Objective To evaluate the antifeedant, insecticidal and growth inhibition activities of Solanum pseudocapsicum (S. pseudocapsicum) seed extracts against Spodoptera litura (S. litura) and Helicoverpa armigera (H. armigera). Methods Hexane, diethyl ether, dichloromethane and ethyl acetate seed extracts were prepared and tested for antifeedant, insecticidal and growth inhibitory activities against fourth instar larvae of S. litura and H. armigera. Results Ethyl acetate extract showed promising antifeedant and insecticidal activities against S. litura and H. armigera. Percentage of deformed larvae, pupae and adults were maximum in treatment of ethyl acetate extract. Percentage of successful adult emergence was deteriorated by seeds on extract treated larvae. Conclusions Ethyl acetate extracts of S. pseudocapsicum, showed higher efficiency of antifeedant, insecticidal and growth inhibition activities. Hence, it can be used to controll agricultural insect pests, S. litura and H. armigera.

992–998

Authors: Urmila M Aswar, Padmaja P Kalshetti, Suhas M Shelke, Sharad H Bhosale, Subhash L Bodhankar

Objective To study the derivatives of 1,2,4-triazino[5,6-b]indole-3-thione for antidepressant activity in olfactory bulbectomized (OBX) rats. Out of various derivatives tested for acute tail suspension test, the two derivatives showing prominent action were selected for bilateral olfactory bulbectomy model of chronic depression in rats. Methods The sub acute effects of 14-day oral pretreatment of two derivatives labeled as 3a (70 mg/kg) and 3r (70 mg/kg), imipramine (20 mg/kg), fluoxetine (30 mg/kg) and moclobemide (15 mg/kg) were evaluated on bilateral bulbectomy induced rise in body weight, hyperphagia, hyperactivity, and on sexual dysfunction. The serum sodium concentration, body temperature, and heart rate were also recorded. Results The derivatives 3a and 3r showed reversal of drop in body weight, reversed OBX induced hyperactivity, normalized body temperature, heart rate, and serum sodium concentration. In elevated maze test, moclobemide, 3a, 3r treatment significantly reduced time spent in open arm as compared to OBX rats. 3a and 3r also improved sexual behavior parameters. Conclusions The present study shows promising antidepressant action and provides a proof of concept for the chronic treatment of 3a, 3r to treat depression.

1002–1008

Authors: Priyanka Soni, Anees Ahmad Siddiqui, Jaya Dwivedi, Vishal Soni

India has a great wealth of various naturally occurring plant drugs which have great potential pharmacological activities. Datura stramonium (D. stramonium) is one of the widely well known folklore medicinal herbs. The troublesome weed, D. stramonium is a plant with both poisonous and medicinal properties and has been proven to have great pharmacological potential with a great utility and usage in folklore medicine. D. stromonium has been scientifically proven to contain alkaloids, tannins, carbohydrates and proteins. This plant has contributed various pharmacological actions in the scientific field of Indian systems of medicines like analgesic and antiasthmatic activities. The present paper presents an exclusive review work on the ethnomedical, phytochemical, pharmacological activities of this plant.