Asian Pacific Journal of Tropical Biomedicine

589-596

Authors: Morteza Izadi, Arman Is’haqi, Mohammad Ali Is’haqi, Nematollah Jonaidi Jafari, Fatemeh Rahamaty, Abdolali Banki

Nervous system infections are among the most important diseases in travellers. Healthy travellers might be exposed to infectious agents of central nervous system, which may require in-patient care. Progressive course is not uncommon in this family of disorders and requires swift diagnosis. An overview of the available evidence in the field is, therefore, urgent to pave the way to increase the awareness of travel-medicine practitioners and highlights dark areas for future research. In November 2013, data were collected from PubMed, Scopus, and Web of Knowledge (1980 to 2013) including books, reviews, and peer-reviewed literature. Works pertained to pre-travel care, interventions, vaccinations related neurological infections were retrieved. Here we provide information on pre-travel care, vaccination, chronic nervous system disorders, and post-travel complications. Recommendations with regard to knowledge gaps, and state-of-the-art research are made. Given an increasing number of international travellers, novel dynamic ways are available for physicians to monitor spread of central nervous system infections. Newer research has made great progresses in developing newer medications, detecting the spread of infections and the public awareness. Despite an ongoing scientific discussion in the field of travel medicine, fu rther research is required for vaccine development, state-of-the-art laboratory tests, and genetic engineering of vectors.

598-602

Authors: Kanungnit Congpuong, Rungniran Sukaram, Yuparat Prompan, Aibteesam Dornae

Objective: To study the genetic diversity at the msp-1, msp-2, and glurp genes of Plasmodium falciparum (P. falciparum) isolates from 3 endemic areas in Thailand: Tak, Kanchanaburi and Ranong provinces. Methods: A total of 144 P. falciparum isolates collected prior to treatment during January, 2012 to June, 2013 were genotyped. DNA was extracted; allele frequency and diversity of msp-1, msp-2, and glurp genes were investigated by nested polymerase chain reaction. Results: P. falciparum isolates in this study had high rate of multiple genotypes infection (96.5%) with an overall mean multiplicity of infection of 3.21. The distribution of allelic families of msp-1 was significantly different among isolates from Tak, Kanchanaburi, and Ranong but not for the msp-2. K1 and MAD20 were the predominant allelic families at the msp-1 gene, whereas alleles belonging to 3D7 were more frequent at the msp-2 gene. The glurp gene had the least diverse alleles. Population structure of P. falciparum isolates from Tak and Ranong was quite similar as revealed by the presence of similar proportions of MAD20 and K1 alleles at msp-1 loci, 3D7 and FC27 alleles at msp-2 loci as well as comparable mean MOI. Isolates from Kanchanaburi had different structures; the most prevalent alleles were K1 and RO33. Conclusions: The present study shows that P. falciparum isolates from Tak and Ranong provinces had similar allelic pattern of msp-1 and msp-2 and diversity but different from Kanchanaburi isolates. These allelic variant profiles are valuable baseline data for future epidemiological study of malaria transmission and for continued monitoring of polymorphisms associated with antimalarial drug resistance in these areas.

610-613

Authors: Sidgi Syed Anwer Hasson, Mohammed Saeed Al-Balushi, Muzna Hamed Al Yahmadi, Juma Zaid Al-Busaidi, Elias Antony Said, Mohammed Shafeeq Othman, Talal Abdullah Sallam, Mohammed Ahmad Idris, Ali Abdullah Al-Jabri

Objective: To investigate the levels of zinc-α-2-glycoprotein (ZAG) among Omani AIDS patients receiving combined antiretroviral therapy (cART). Methods: A total of 80 Omani AIDS patients (45 males and 35 females), average age of 36 years, who were receiving cART at the Sultan Qaboos University Hospital (SQUH), Muscat, Oman, were tested for the levels of ZAG. In addition, 80 healthy blood donors (46 males and 34 females), average age of 26 years, attending the SQUH Blood Bank, were tested in parallel as a control group. Measurement of the ZAG levels was performed using a competitive enzyme-linked immunosorbent assay and in accordance with the manufacturer’s instructions. Results: The ZAG levels were found to be significantly higher among AIDS patients compared to the healthy individuals (P=0.033). A total of 56 (70%) of the AIDS patients were found to have higher levels of ZAG and 16 (20%) AIDS patients were found to have high ZAG levels, which are significantly (P>0.031) associated with weight loss. Conclusions: ZAG levels are high among Omani AIDS patients on cART and this necessitates the measurement of ZAG on routine basis, as it is associated with weight loss.

618-626

Authors: Sahar Y. Al-Okbi, Doha A. Mohamed, Thanaa E. Hamed, Reham SH. Esmail, Souria M. Donya

Objective: To investigate the protective effect of extracts prepared from avocado, walnut, flaxseed and Eruca sativa seeds in a rat model of kidney dysfunction induced by intraperitoneal cisplatin. Methods: Ethanol and petroleum ether extracts mixture was prepared from each plant. Six groups of rats were conducted; control healthy, cisplatin group and four test groups where rats were given daily oral dose of each extract mixture before cisplatin injection. Different biochemical and cytogenetic parameters and kidney histopathology were determined. Acute toxicity was tested fo r the nutraceuticals. Total phenolic contents, fatty acids (FA) and unsaponifiable matter were assessed in the extracts. Results: Walnut ethanol extract showed the highest content of total phenolic. FA analysis revealed that all the studied plants were rich in unsaturated FA. Gas-liquid chromatographic investigation of the unsaponifiable matter showed the presence of campesterol, stigmasterol and β-sitosterol in all the studied plants. Cisplatin treatment induced significant increase in plasma urea, creatinine and malondialdehyde along with significant reduction of plasma albumin, total protein, catalase and total antioxidant as well as reduction in creatinine clearance. Histopathological examination proved the induction of kidney dysfunction. Some sorts of chromosomal aberration and spermshape abnormalities were noticed after cisplatin treatment. Administration of extracts mixtures produced improvements in biochemical, histopathological and cytogenetic parameters. Conclusions: Administration of the studied nutraceuticals proved to possess protective role against cisplatin-induced nephrotoxicity, chromosomal aberration and abnormal sperms. All studied nutraceuticals showed complete safety.

633-641

Authors: Hira Fahim, Javid Iqbal Dasti, Ihsan Ali, Safia Ahmed, Muhammad Nadeem

Objective: To evaluate physico-chemical properties and antimicrobial potential of indigenous honey samples against different reference strains including Escherichia coli ATCC 8739, Enterobacter aerogenes ATCC 13048, Pseudomonas aeroginosa ATCC 9027, Bacillus subtilis ATCC 6633, Staphylococcus aureus ATCC 25923, Salmonella typhi ATCC 14028, Klebsiella pneumonia ATCC 13883, Aspergillus niger ATCC 16404, Rhizopus oligosporus PCSIR1, Candida albicans ATCC 14053 and Candida utilis ATCC 9950. Methods: By using standard methods samples were evaluated for their antimicrobial properties including additive effect of starch and non-peroxidase activity, antioxidative properties (phenol contents, flavonoid contents, 1,1-diphenyl-2-picrylhydrazyl radical scavenging activity). Prior to this evaluation, complete physico-chemical properties including pH, color, ash contents, protein contents, moisture contents, hydroxymethyl furfural contents, total sugar contents, reducing sugar and non-reducing sugar contents were analyzed. Results: Relatively higher ash contents were found in the Siddar honey i.e. (0.590 0±0.033 6)% and small honey showed relatively higher protein contents i.e. (777.598±9.880) mg/kg. The moisture contents of tested honey samples ranged between 13.8%-16.6%, total sugar contents from 61.672%- 72.420% and non-reducing sugar contents from 1.95%-3.93%. Presences of phenolic contents indicate higher antioxidant potential of these honey samples. All bacteria showed clear inhibition zones in response to tested honey samples whereas fungi and yeast showed inhibition at higher concentrations of these honey samples. For Escherichia coli, Bacillus subtilis, Salmonella typhi, Pseudomonas aeroginosa and Aspergillus niger, overall the small honey showed the higher activity than other honey samples. Conclusion: Physico-chemical analysis of honey samples confirmed good quality of honey according to the standards set by European Union Commission and Codex Alimentarius Commission. Evaluation of these honey samples confirms antimicrobial potential of particular types of honeys indigenous to Pakistan.

650-654

Authors: Shaheen Faizi, Saima Sumbul, Muhammed Ali Versiani, Rubeena Saleem, Aisha Sana, Hira Siddiqui

Objective: To explore the phytochemical constituents from petroleum ether and dichloromethane extracts of Moringa oleifera (M. oleifera) roots using GC/GC-MS. Methods: A total of 5.11 kg fresh and undried crushed root of M. oleifera were cut into small pieces and extracted with petroleum ether and dichloromethane (20 L each) at room temperature fo r 2 d. The concentrated extracts were subjected to their GC-MS analysis. Results: The GC-MS analysis of the petroleum ether and dichloromethane extracts of M. oleifera roots, which showed promising biological activities, has resulted in the identification 102 compounds. These constituents belong to 15 classes of compounds including hydrocarbons, fatty acids, esters, alcohols, isothiocyanate, thiocyanate, pyrazine, aromatics, alkamides, cyanides, steroids, halocompounds, urea and N-hydroxyimine derivatives, unsaturated alkenamides, alkyne and indole. GC/GC-MS studies on petroleum ether extract of the roots revealed that it contained 39 compounds, belonging to nine classes. Cyclooctasulfur S8 has been isolated as a pure compound fr om the extract. The major compounds identified from petroleum ether extract were trans- 13-docosene (37.9%), nonacosane (32.6%), cycloartenol (28.6%) nonadecanoic acid (13.9%) and cyclooctasulfur S8 (13.9%). Dichloromethane extract of the roots was composed of 63 compounds of which nasimizinol (58.8%) along with oleic acid (46.5%), N-benzyl-N-(7-cyanato heptanamide (38.3%), N-benzyl-N-(1-chlorononyl) amide (30.3%), bis [3-benzyl prop-2-ene]-1-one (19.5%) and N, N-dibenzyl-2-ene pent 1, 5-diamide (11.6%) were the main constituents. Conclusions: This study helps to predict the formula and structure of active molecules which can be used as drugs. This result also enhances the traditional usage of M. oleifera which possesses a number of bioactive compounds.

659-662

Authors: Bamba Sanata, Ouédraogo Abdoul Salam, Sangaré Ibrahim, Zida Adama, Cissé Mamoudou, Karou D. Simplice, Simpore Jacques, Guiguemdé T. Robert, Hennequin Christophe

Objective: To identify Candida species in asymptomatic subjects in Bobo-Dioulasso (Burkina Faso) by the matrix-assisted laser desorption ionization-time of flight mass spectrometry. Methods: A cross-sectional study was conducted from January to February 2013 in BoboDioulasso to collect fecal and urine specimens from voluntary donors. Fungal strains were isolated on Sabouraud dextrose agar and analyzed using matrix-assisted laser desorption ionisationtime-of-flight mass spectrometry. Results: A total of 135 samples including stools (78.5%, 106/135) and urine (21.5%, 29/135) were analyzed. The results revealed that fecal specimens contained mainly Candida krusei (C. krusei) (42.5%) followed by Candida albicans (29.3%), Candida glabrata (18.0%) and Candida tropicalis (C. tropicalis) (4.7%). C. krusei (34.6%) was also found to be the most frequently identified in urine samples followed by Candida albicans (27.0%), C. tropicalis (15.4%) and Candida parapsilosis. However, uncommon species such as Candida nivariensis, Candida kefyr, Candida norvegensis, Candida parapsilosis, Candida lusitaniae and Candida robusta were also identified from fecal and urines samples. Conclusions: This study noted the emergence of species such as C. krusei, Candida glabrata, Candida parapsiolosis, C. tropicalis, Candida nivariensis, Candida norvegensis, and others. It is an imperative to take into account the existence of these species in the therapeutic management of patients in Bobo-Dioulasso.

669-672

Authors: Fahmi Yousef Khan, Mohammed Abukhattab, Mohammed AbuKamar, Deshmukh Anand

Objective: To describe the clinical presentation, underlying diseases, antimicrobial susceptibility, treatment and outcome of Klebsiella pneumoniae meningitis patients. Methods: This retrospective study involved all patients with 15 years of age or older who admitted to Hamad General Hospital with culture proven Klebsiella pneumoniae meningitis from January 1, 2007 to December 31, 2012. Results: A total of ten cases were identified (nine males and one female). Their mean age was (43.3±12.8) years. Eight patients (80%) had nosocomial meningitis with neurosurgery being the most frequent associated condition. Fever and altered consciousness were the most frequent symptom. Cerebrospinal fluid showed elevated protein and glucose levels. Gram stain showed Gram-negative rods in 50% of cases, while positive cerebrospinal fluid culture results were found in all patients. Multidrug resistance was observed in two cases, and all patients had received appropriate empirical and definitive antibiotic treatments. The mean duration of intravenous antimicrobial treatment was (19.3±7.0) d and all patients with external ventricular drains underwent removal of the device, while in-hospital mortality was 50%. Conclusions: The number of cases was too small to come up with therapeutic and prognostic conclusions. Further large-scale prospective study is needed.

505-514

Authors: Elif Burcu Bali, Leyla Açık, Gülçin Akca, Meral Sarper, Mualla Pınar Elçi, Ferit Avcu, Mecit Vural

Objective: To investigate the in vitro antimicrobial potential of Thermopsis turcica Kit Tan, Vural & Küçüködük against periodontopathogenic bacteria, its antioxidant activity and cytotoxic effect on various cancer cell lines. Methods: In vitro antimicrobial activities of ethanol, methanol, ethyl acetate (EtAc), n-hexane and water extracts of Thermopsis turcica herb against periodontopathogenic bacteria, Aggregatibacter actinomycetemcomitans ATCC 29523 and Porphyromonas gingivalis ATCC 33277 were tested by agar well diffusion, minimum inhibitory concentration (MIC) and minimal bactericidal concentration (MBC). Antioxidant properties of the extracts were evaluated by 1,1-diphenyl-2-picryl-hydrazyl radical scavenging activity and β-carotene bleaching methods. Amounts of phenolic contents of the extracts were also analysed by using the Folin-Ciocalteu reagent. Additionally, cytotoxic activity of the extracts on androgen-insensitive prostate cancer, androgen-sensitive prostate cancer, chronic myelogenous leukemia and acute promyelocytic leukemia human cancer cell lines were determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Human gingival fibroblast cells were used as a control. Results: Our data showed that EtAc extract had the highest antimicrobial effect on Aggregatibacter actinomycetemcomitans (MIC: 1.562 mg/mL, MBC: 3.124 mg/mL) and Porphyromonas gingivalis (MIC: 0.781 mg/ mL, MBC: 1.562 mg/mL). In antioxidant assays, EtAc extract exhibited also the highest radical scavenging activity [IC50=(30.0±0.3) μg/mL] and the highest inhibition [(74.35±0.30)%] against lineloic acide oxidation. The amount of phenolic content of it was also the highest [(162.5±1.2) μg/mg gallic acid]. In cytotoxic assay, only ethanol [IC50=(80.00±1.21) μg/mL] and EtAc extract [IC50=(70.0±0.9) μg/mL] were toxic on acute promyelocytic leukemia cells at 20-100 μg/mL (P<0.05). However, no toxic effect was observed on human gingival fibroblast cells. Conclusions: According to our findings, owing to its antioxidant and cytotoxic potential, EtAc extract might include anticancer agents for acute promyelocytic leukemia.

520-522

Authors: David Zant, Daniel A. Gubler

Objective: To identify the major organic compounds of Artemisia australis (A. australis), a plant used in traditional Hawaiian medicine for the treatment of asthma. Methods: The dichloromethane extract of A. australis was analyzed by gas chromatographymass spectroscopy and major compounds were identified by a National Institute of Standards and Technology library search and confirmed by peak enhancement. Results: The major chemical components of A. australis include eucalyptol, borneol, and caryophyllene. Conclusions: The presence and biological activity of eucalyptol correlate very well with the usage of this plant in traditional Hawaiian medicine.

529-537

Authors: Eun-Jin Yang, Ji-Young Moon, Sang Suk Kim, Kyong-Wol Yang, Wook Jae Lee, Nam Ho Lee, Chang-Gu Hyun

Objective: To investigate the anti-inflammatory effects of Jeju seaweeds on macrophage RAW 264.7 cells under lipopolysaccharide (LPS) stimulation. Methods: Ethyl acetate fractions were prepared from five different types of Jeju seaweeds, Dictyopteris divaricata (D. divaricata), Dictyopteris prolifera (D. prolifera), Prionitis cornea (P. cornea), Grateloupia lanceolata (G. lanceolata), and Grateloupia filicina (G. filicina). They were screened for inhibitory effects on proinflammatory mediators and cytokines such as nitric oxide (NO), prostaglandin E2, tumor necrosis factor-α (TNF-α), and interleukin-6 (IL-6). Results: Our results revealed that D. divaricata, D. prolifera, P. cornea, G. lanceolata, and G. filicina potently inhibited LPS-stimulated NO production (IC50 values were 18.0, 38.36, 38.43, 32.81 and 37.14 µg/mL, respectively). Consistent with these findings, D. divaricata, D. prolifera, P. cornea, and G. filicina also reduced the LPS-induced and prostaglandin E2 production in a concentration-dependent manner. Expectedly, they suppressed the expression of inducible NO synthase and cyclooxygenase-2 at the protein level in a dose-dependent manner in the RAW 264.7 cells, as determined by western blotting. In addition, the levels of TNF-α and IL-6, released into the medium, were also reduced by D. divaricata, D. prolifera, P. cornea, G. lanceolata, and G. filicina in a dose-dependent manner (IC50 values for TNF-α were 16.11, 28.21, 84.27, 45.52 and 74.75 µg/mL, respectively; IC50 values for IL-6 were 37.35, 80.08, 103.28, 62.53 and 84.28 µg/mL, respectively). The total phlorotannin content was measured by the Folin-Ciocalteu method and expressed as phloroglucinol equivalents. The content was 92.0 µg/mg for D. divaricata, 151.8 µg/ mg for D. prolifera, 57.2 µg/mg for P. cornea, 53.0 µg/mg for G. lanceolata, and 40.2 µg/mg for G. filicina. Conclusions: Thus, these findings suggest that Jeju seaweed extracts have potential therapeutic applications for inflammatory responses.

545-548

Authors: Nurfariza Ahmad Roslen, Nur Aizura Mat Alewi, Hadji Ahamada, Mohammad Syaiful Bahari Abdull Rasad

Objective: To screen the cytotoxic activity of Melastoma malabathricum (M. malabathricum) against human breast cancer cell line (MCF-7) in vitro. Methods: A three steps extraction protocol using n-hexane, chloroform and methanol as the solvents systems was carried out on leaves, stems and flowers of M. malabathricum. Dimethyl sulfoxide was used in extracts dilution and serial dilutions were conducted to obtain five different extract concentrations (100 µg/mL, 50 µg/mL, 25 µg/mL, 12.5 µg/mL and 6.25 µg/mL). The evaluation of cell growth was determined using methylene blue assay. Results: Methanol extract from the leaves showed significant anticancer activity against MCF-7 cell lines with the IC 50 value of 7.14 µg/ml while methanol and chloroform extract from the flowers exhibited a moderate activity towards MCF-7 cell line with the IC50 value of 33.63 µg/mL and 45.76 µg/mL respectively after 72 h of treatment. Conclusions: The extracts from leaves and flowers of M. malabathricum showed promising anticancer activity toward human breast cancer cell lines with the lowest IC50 at 7.14 µg/mL while the extracts from stems showed less growth inhibition activity.

557-560

Authors: Sylvia Joanne, Indra Vythilingam, Nava Yugavathy, Jonathan Inbaraj Doss

Objective: To develop an artificial and modified Wolbachia removal technique using tetracycline fr om naturally Wolbachia infected Aedes albopictus (Ae. albopictus) so as to be able to produce generations of Wolbachia free offsprings. Methods: In this study, seven different tetracycline treatment methods were conducted to obtain the best removal method. Four methods focused on larvae tetracycline treatment, one method on both larvae and adult tetracycline treatment and the last two methods on adult mosquito sucrose treatment. Results: All larval tetracycline treatments resulted in either high larvae mortality, sterile F0 adult mosquitoes or unsuccessful Wolbachia removal. Treatment of both larvae and adults resulted in reduced larvae mortality, successful Wolbachia removal but slow mosquito fecundity. As for the adult treatment, 1.0 mg/mL as previously published was not able to completely remove Wolbachia in F 1 generation whereas 1.25 mg/mL successfully removed Wolbachia from F1 and F2 mosquitoes in 2 weeks. Conclusions: This method is different from the previously published methods as it provides an improved Wolbachia removal technique from Ae. albopictus with high egg hatchability, low larvae mortality, increased fecundity and better Wolbachia removal rate.

566-569

Authors: Bahador Sarkari, Asgari Qasem, Mohammad Reza Shafaf

Objective: To assess knowledge, attitudes, and practices (KAP) of inhabitants of an endemic area in Fars province toward cutaneous leishmaniasis (CL). Methods: The study was carried out in Lapui district in Fars province, south of Iran, one of the most important foci of CL in this province. Sample size (237 residents) was calculated based on population. House-to-house survey was done to collect the data regarding knowledge, attitudes, and practices of the inhabitants. The head of each household was interviewed by a trained staff to assess his/her KAP related to CL. A semi-structured KAP questionnaire was used for data collection. Results: Mean age of participants was 39 and more than half of the respondents were in the age group of 31-40. Males constituted 172 (72.5%) of subjects. Most of the respondents (84.3%) were literate. The majority of the study population (83%) had heard about Salak (local name for CL) and most of these respondents (91%) were aware that CL is presented with a cutaneous lesion. Nearly two-third of the participants (63.5%) stated the bite of mosquito (not specifically sandflies) for CL transmission. The respondents’ attitude regarding the treatment of CL was not satisfactory since only 48% believed that CL can be treated by medicine. A noticeable proportion of respondents (21%) believed in indigenous medicine for the treatment of CL. A small proportion of respondents (14%) stated that traditional healers are good at treating this disease. More than two-third (69%) of respondents believed that the disease is preventable although most of interviewees did not know about preventive measures. Conclusions: In this study, insufficient knowledge of community about infection nature, vector, transmission mode and preventive measures of CL, highlights the needs for a health education initiative to enhance the awareness of people about CL. This would improve inhabitants’ contribution in control program of CL in this area.

575-580

Authors: Bhesh Raj Sharma, Dong Young Rhyu

Objective: To evaluate anti-diabetic effect of Caulerpa lentillifera (C. lentillifera). Methods: The inhibitory effect of C. lentillifera extract on dipeptidyl peptidase-IV and α-glucosidase enzyme was measured in a cell free system. Then, interleukin-1β and interferon-γ induced cell death and insulin secretion were measured in rat insulinoma (RIN) cells by 3-(4,5- dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and ELISA kit, respectively. Glucose uptake and glucose transporter expression were measured by fluorometry and western blotting, using 3T3-L1 adipocytes. Results: C. lentillifera extract significantly decreased dipeptidyl peptidase-IV and α-glucosidase enzyme activities, and effectively inhibited cell death and iNOS expression in interleukin-1β and interferon-γ induced RIN cells. Furthermore, C. lentillifera extract significantly enhanced insulin secretion in RIN cells and glucose transporter expression and glucose uptake in 3T3-L1 adipocytes. Conclusions: Thus, our results suggest that C. lentillifera could be used as a potential antidiabetic agent.

586-588

Authors: Azari Omid, Ali Asghar Mozaffari

A five-year-old cow was referred to the veterinary faculty hospital for treatment of a swelling mass with a cutaneous fistula at the left lower part of the chest wall, between 7th till 9th intercostals space. Abdominal pain in palpation of the mass was observed with no abnormality in clinical symptoms. In surgical exploration, skin incision was carried out on the swelling mass and surprisingly a sinus tract with a sharp metallic rod (26 cm length) that continued to the reticulum lumen was identified. Because of unsuccessful attempting to remove of the foreign body, flank laparotomy and rumenotomy was performed. After one month, the cow led to complete recovery.

421-429

Authors: Erhabor Osaro, Njemanze Chima

The National Health Service (NHS) is a term used to describe the publicly funded healthcare delivery system providing quality healthcare services in the United Kingdom. There are several challenges militating against the effective laboratory service delivery in the NHS in England. Biomedical scientists work in healthcare to diagnose disease and evaluate the effectiveness of treatment through the analysis of body fluids and tissue samples from patients. They provide the “engine room” of modern medicine with 70% of diagnosis based on the laboratory results generated by them. This review involved the search of literature for information on working condition of biomedical scientist in the NHS in England. Laboratory service delivery in the NHS in England faces numerous daunting challenges; staffing levels in the last few years have become dangerously low, less remunerated, relatively less experienced and predominantly band 5’s, multidisciplinary rather than specialty based, associated with working more unsocial hours without adequate recovery time, de-banding of staff, high staff turnaround, profit and cost driven rather than quality. These factors has resulted in burn out, low morale, high sickness absences, increased error rate, poor team spirit, diminished productivity and suboptimal laboratory service delivery. There is the urgent need to retract our steps on unpopular policies to ensure that patient care is not compromised by ensuring adequate staffing level and mix, ensuring adequate remuneration of laboratory staff, implementing evidenced-based specialty oriented service, determining the root cause/s for the high staff turnover and implementing corrective action, identifying other potential sources of waste in the system rather than pruning the already dangerously low staffing levels and promoting a quality delivery side by side cost effectiveness.

436-440

Authors: Algenes Aranha, Usman H Malabu, Venkat Vangaveti, Elham Saleh Reda, Yong Mong Tan, Kunwarjit Singh Sangla

Objective: To look into the glucose tolerance test characteristics and determine complications in non-gestational diabetes pregnant subjects. Methods: From 2006 to 2009 all non-gestational diabetes mellitus (non-GDM) pregnant women who delivered macrosomia at the North Australia’s Townsville Hospital were retrospectively reviewed by extracting data from clinical record. Glucose tolerance tests results were analysed in the light of an earlier diagnosis of non-GDM. Results: Ninety-one non-GDM mothers with macrosomia were studied and compared with 41 normoglycemic subjects without macrosomia. Of the subjects with non-GDM macrosomia, 45 (49.4%) had normal 50 g glucose challenge test (GCT) without further testing, another 8 (8.8%) had abnormal GCT but normal 75 g oral glucose tolerance test (OGTT). A total of 4 (4.4%) subjects had normal GCT and OGTT. Interestingly, 14 out of 16 (87.5%) subjects who were tested with OGTT owing to past history of macrosomia had normal results but delivered macrosomic babies. Only 12 subjects had both GCT and OGTT, the rest of the cohort had either of the two tests. Subjects with non-GDM macrosomia had higher frequency of neonatal hypoglycaemia 34% as compared to 10% in nonmacrosomic babies (P=0.003). Other feto-maternal complications were similar in both groups. Conclusions: No significant pattern of glucose tolerance characteristics was identified in nonGDM mothers with macrosomic babies. In spite of being normoglycemic significant neonatal hypoglycaemia was recorded in non-GDM macrosomic babies. Further prospective studies on a larger population are needed to verify our findings.

446-450

Authors: Lim Boon Huat, Alfonso Olivos Garcia, Tan Zi Ning, Wong Weng Kin, Rahmah Noordin, Siti Shafiqah Anaqi Azham, Lee Zhi Jie, Guee Cher Ching, Foo Phiaw Chong, Pim Chau Dam

Objective: To characterize the Entamoeba histolytica (E. histolytica) antigen(s) recognized by moribound amoebic liver abscess hamsters. Methods: Crude soluble antigen of E. histolytica was probed with sera of moribund hamsters in 1D- and 2D-Western blot analyses. The antigenic protein was then sent for tandem mass spectrometry analysis. The corresponding gene was cloned and expressed in Escherichia coli BL21-AI to produce the recombinant E. histolytica ADP-forming acetyl-CoA synthetase (EhACS) protein. A customised ELISA was developed to evaluate the sensitivity and specificity of the recombinant protein. Results: A ~75 kDa protein band with a pI value of 5.91-6.5 was found to be antigenic; and not detected by sera of hamsters in the control group. Tandem mass spectrometry analysis revealed the protein to be the 77 kDa E. histolytica ADP-forming acetyl-CoA synthetase (EhACS). The customised ELISA results revealed 100% sensitivity and 100% specificity when tested against infected (n=31) and control group hamsters (n=5) serum samples, respectively. Conclusions: This finding suggested the significant role of EhACS as a biomarker for moribund hamsters with acute amoebic liver abscess (ALA) infection. It is deemed pertinent that future studies explore the potential roles of EhACS in better understanding the pathogenesis of ALA; and in the development of vaccine and diagnostic tests to control ALA in human populations.

456-462

Authors: Nagwa Mohamed Ammar, Mohammed Said Hefnawy, Sahar Youssef Al-Okbi, Doha Abdou Mohamed, Nabil Khamis El-Sayed, Amira Ahmed El -Anssary, Tom Mabry

Objective: To study the antioxidant and anti-inflammatory activity of Butia capitata (B. capitata) leaf extracts along with phytochemical analysis of the proposed bioactive constituents. Methods: Different successive extracts of B. capitata Becc. leaves were prepared with selective organic solvents and screened for their anti-inflammatory activities in tested animals and invitro antioxidant effect. An extensive phytochemical investigation of the bioactive extracts through paper chromatography, thin layer chromatography, column chromatography, gasliquid chromatography (GLC), high pressure liquid chromatography and spectral analysis. GCMass, ultraviolet, hydrogen and carbon nuclear magnetic resonance, electron ionization-mass spectrometry, heteronuclear multiple bond correlation and heteronuclear multiple quantum correlation were carried out. Results: Results showed that different extracts possess promising antioxidant effect and significant anti-inflammatory activity with variable degrees. The results of the phytochemical investigation of the bioactive extracts revealed the presence of volatile substances, lipoidal matter, α-tocopherol, free sugars, polysaccharides and flavonoidal compounds. Conclusions: B. capitata leaf extracts were shown to possess variable antioxidant effect, the most promising was methanol extract. Both polar and non polar extracts were proved to have anti-inflammatory activity, the non polar extract was superior in this respect. The bioactivity of the extracts was ascribed to the presence of flavonoids, sterols and α-tocopherol.

473-479

Authors: Nizam Uddin, Md. Rakib Hasan, Md. Monir Hossain, Arjyabrata Sarker, A.H.M. Nazmul Hasan, A.F.M. Mahmudul Islam, Mohd. Motaher H. Chowdhury, Md. Sohel Rana

Objective: To investigate the therapeutic effects of methanol extract of Citrus macroptera Montr. fruit in α-amylase inhibitory activity (in vitro) and hypoglycemic activity in normal and glucose induced hyperglycemic rats (in vivo). Methods: Fruits of Citrus macroptera without rind was extracted with pure methanol following cold extraction and tested for presence of phytochemical constituents, α-amylase inhibitory activity, and hypoglycemic effect in normal rats and glucose induced hyperglycemic rats. Results: Presence of saponin, steroid and terpenoid were identified in the extract. The results showed that fruit extract had moderate α-amylase inhibitory activity [IC50 value=(3.638±0.190) mg/mL] as compared to acarbose. Moreover at 500 mg/kg and 1 000 mg/kg doses fruit extract significantly (P<0.05 and P<0.01 respectively) reduced fasting blood glucose level in normal rats as compared to glibenclamide (5 mg/kg). In oral glucose tolerance test, 500 mg/kg dose significantly reduced blood glucose level (P<0.05) at 2 h but 1 000 mg/kg dose significantly reduced blood glucose level at 2 h and 3 h (P<0.05 and P<0.01 respectively) whereas glibenclamide (5 mg/ kg) significantly reduced glucose level at every hour after administration. Overall time effect is also considered extremely significant with F value=23.83 and P value=0.0001 in oral glucose tolerance test. Conclusion: These findings suggest that the plant may be a potential source for the development of new oral hypoglycemic agent.

486-491

Authors: Pradeep Singh, Ratan Lal Khosa, Shruti Srivastava, Garima Mishra, Keshri Kishor Jha, Sourabh Srivastava, Sangeeta, Ramesh Kumar Verma, Mohd Adil Tahseen

Objective: To evaluate the diagnostic pharmacognostical characters of Costus speciosus (aerial parts) along with their physico-chemical parameters and fluorosence analysis. Method: The pharmacognostical characters were determined in terms of macroscopy, microscopy, powder microscopy, leaf constant, fluorescence analysis and preliminary phytochemical investigation. Results: The findings of macroscopy revealed that leaves elliptic to oblong or oblong-lancoelate, thick, spirally arranged, with stem clasping sheaths up to 4 cm, flowers large, white, conelike terminal spikes, with bright red bracts. Transverse section of leaflet showed the presence of cuticularised epidermis with polygonal cells on adaxial surface and bluntly angled cells on abaxial surface of lamina, mesophyll cells differentiated in to single layered palisade cells on each surface and 2-3 layered spongy parenchyma, unicellular and uniseriate multicellular covering trichomes, paracytic stomata and vascular bundles surrounded by sclerenchymatous multicellular sheath. Preliminary phytochemical screening exhibited the presence of various phytochemical groups like alkaloids, glycosides, steroids, phenolic constituents. Further, the leaf constants, powder microscopy and fluorescence characteristics indicated outstanding results from this investigation Conclusions: Various pharmacognostical and physico-chemical parameters have pivotal roles in identification, authentication and establishment of quality parameters of the species.

498-504

Authors: Pascalina Chanda-Kapata, Emmanuel Chanda, Freddie Masaninga, Annette Habluetzel, Felix Masiye, Ibrahima Soce Fall

Objective: To establish the appropriateness of malaria case management at health facility level in fo ur districts in Zambia. Methods: This study was a retrospective evaluation of the quality of malaria case management at health facilities in four districts conveniently sampled to represent both urban and rural settings in different epidemiological zones and health facility coverage. The review period was fr om January to December 2008. The sample included twelve lower level health facilities from fo ur districts. The Pearson Chi-square test was used to identify characteristics which affected the quality of case management. Results: Out of 4891 suspected malaria cases recorded at the 12 health facilities, more than 80% of the patients had a temperature taken to establish their fever status. About 67% (CI95 66.1-68.7) were tested for parasitemia by either rapid diagnostic test or microscopy, whereas the remaining 22.5% (CI95 21.3.1-23.7) were not subjected to any malaria test. Of the 2247 malaria cases reported (complicated and uncomplicated), 71% were parasitologically confirmed while 29% were clinically diagnosed (unconfirmed). About 56% (CI95 53.9-58.1) of the malaria cases reported were treated with artemether-lumefantrine (AL), 35% (CI95 33.1-37.0) with sulphadoxine-pyrimethamine, 8% (CI95 6.9- 9.2) with quinine and 1% did not receive any anti-malarial. Approximately 30% of patients WHO were found negative for malaria parasites were still prescribed an anti-malarial, contrary to the guidelines. There were marked inter-district variations in the proportion of patients in WHOm a diagnostic tool was used, and in the choice of anti-malarials for the treatment of malaria confirmed cases. Association between health worker characteristics and quality of case malaria management showed that nurses performed better than environmental health technicians and clinical officers on the decision whether to use the rapid diagnostic test or not. Gender, in service training on malaria, years of residence in the district and length of service of the health worker at the facility were not associated with diagnostic and treatment choices. Conclusions: Malaria case management was characterised by poor adherence to treatment guidelines. The non-adherence was mainly in terms of: inconsistent use of confirmatory tests (rapid diagnostic test or microscopy) for malaria; prescribing anti-malarials which are not recommended (e.g. sulphadoxine-pyrimethamine) and prescribing anti-malarials to cases testing negative. Innovative approaches are required to improve health worker adherence to diagnosis and treatment guidelines.

337-344

Authors: Pongsathon Premratanachai, Chanpen Chanchao

Bee products have long been used in traditional medicine. The raw materials, crude extracts and purified active compounds from them have been found to exhibit interesting bioactivities, such as antimicrobial, anti-inflammatory and antioxidant activities. In addition, they have been widely used in the treatment of many immune-related diseases, as well as in recent times in the treatment of tumors. Bee product peptides induce apoptotic cell death in vitro in several transformed (cancer) human cell lines, including those derived from renal, lung, liver, prostate, bladder and lymphoid cancers. These bioactive natural products may, therefore, prove to be useful as part of a novel targeted therapy for some types of cancer, such as prostate and breast cancer. This review summarizes the current knowledge regarding the in vivo and in vitro potential of selective bee products against tumor cells.

354-357

Authors: Mohammad Khalili, Ehsanollah Sakhaee, Mohammad Reza Aflatoonian, Gholamreza Abdollahpour, Saeed Sattari Tabrizi, Elham Mohammadi Damaneh, Sajad Hossini-nasab

Objective: To evaluate serological findings of bovine leptospirosis which is a zoonotic disease with worldwide distribution caused by Leptospira interrogans. Methods: One hundred and sixty seven sera were collected from 9 commercial dairy herds in jiroft suburbs, from July to October 2011. Microscopic agglutination test (MAT) was used to evaluates serological findings of bovine leptospirosis in Jiroft suburb dairy farms, Kerman province, Iran. Results: Antibodies were found by MAT at least against one serovar of Leptospira interrogans in 29 samples (17.36%) among 167 sera at a dilution 1:100 or higher, and Leptospira pomona was the most prevalent serovar. Positive titers against more than one serovar were detected in 6 sera of the positive samples. Conclusion: This study is the first report of leptospirosis in Southeast Iran and showed that Leptospira pomona was the most and Leptospira icterohaemorrhagiae the least prevalent serovars in Southeast Iran.

362-366

Authors: Kirill Golokhvast, Alexander Sergievich, Nikolay Grigoriev

Objective: To discusse the impact of geophagy on behavior and conditioned-reflex activity of Wistar rats subjected to instrumental stress under experimental conditions. Methods: Experimental geophagy was simulated by adding zeolite-containing tuff (clinoptilolite) to animal feed, the amount relating to 2% of body mass. Tuff was obtained from areas where animals usually eat subsurface rock. Search activity of animals and peculiarities of information and emotional stress were studied through the use of a universal problem chamber. Results: The results of this experimental study showed the negative impact of instrumental stress on laboratory animals, manifested in behavioral dysfunction, in the form of changes in qualitative and quantitative characteristics of search activity. Experimental geophagy contributed to significant improvement in behavioral parameters, confirming the anti-stress effects of the use of natural ingredients. Conclusions: These results suggest that, in natural environmental conditions, "edible" rocks serve as an adaptive tool for recovery from various types of environmental stresses, and are examples of self-medication.

374-378

Authors: Abiodun Falodun, Vincent Imieje, Osayewenre Erharuyi, Ahomafor Joy, Peter Langer, Melissa Jacob, Shabanna Khan, Mohammed Abaldry, Mark Hamann

Objective: To investigate the antileishmanial, antimicrobial and antimalarial activities of the pure metabolites from Jatropha multifida used in African ethnomedicine. Methods: The methanolic stem bark extract of Jatropha multifida used in Nigerian folk medicine as remedy against bacterial infections was subjected to column chromatography and HPLC analyses to obtain three known metabolites, microcyclic lathyrane diterpenoids (1-3). Structures were confirmed by comparison of 1D and 2D spectral data with literature. Results: The three compounds exhibited inhibition of antileishmanial, antimalarial and antimicrobial actions against the tested organisms with compouds 2 and 3 active against Cryptococcus neoformans at IC50 of 8.2 and 8.7 µg/mL, respectively. Conclusions: The research lends support to the ethnomedicinal use of the plant in combating microbial infections, leishmaniasis and malarial infections.

386-392

Authors: Mui-Hung Wong, Li-Fang Lim, Fasihuddin bin Ahmad, Zaini bin Assim

Objective: To investigate antioxidant and antimicrobial activities of two plant species, Litsea elliptica (L. elliptica) and Litsea resinosa (L. resinosa). Methods: In vitro method -2,2-diphenyl-1-picrylhydrazyl radical scavenging assay was conducted for antioxidant activity determination while antimicrobial assay consisted of agar well diffusion assay and mycelial radial growth assay. Results: Methanol extracts of root and stem of L. elliptica and L. resinosa exhibited the highest antioxidant activity with EC50 of 23.99, 41.69, 11.22 and 35.48 mg/L respectively. All methanol extracts of L. resinosa as well as root extracts from L. elliptica showed significant scavenging activity. Hexane extract from stem of L. resinosa presented the largest inhibition zone in Gramnegative bacteria Pseudomonas aeruginosa and Escherichia coli while chloroform extract from inner bark of L. resinosa showed major inhibition towards Gram-positive bacteria Bacillus subtilis. Essential oils from the root of both species showed significant antifungal activities which are 80.11% and 66.85% respectively. Conclusions: Overall, methanol extracts from root and stem of both species showed antioxidant activity comparable to standard butylated hydroxytoluene. Extracts from L. resinosa demonstrated stronger antimicrobial properties compared to that from L. elliptica.

399-403

Authors: Saleem Bani Hani, Mekki Bayachou

Objective: To investigate the role of water-soluble extract of Salvia fruticosa (Greek sage) (S. fruticosa) leaves in reducing both intrinsic cellular and H2O2-induced DNA oxidation in cultured human embryonic kidney 293 cells. S. fruticosa, native to the Eastern-Mediterranean basin, is widely used as a medicinal herb for treatment of various diseases. Methods: Dried leaves of S. fruticosa were extracted in phosphate buffer saline and purified using both vacuum and high pressure filtrations. Each mL of the preparation contained (7.1±1.0) mg of extract. HEK-293 cells were incubated in one set with S. fruticosa extract in the presence of 0.1 mmol/L H 2O2, and in the other set with the addition of the extract alone. The DNA oxidation was measured using fluorescence upon fluorescein isothiocyanate derivatization of 8-oxoguanine moieties. The fluorescence was measured using flow cytometry technique. Results: Cells incubated 3 h with 150 µL extract and exposed to 0.1 mmol/L H2O2 showed lower intensity of fluorescence, and thus lower DNA oxidation. Moreover, cells incubated 3 h with 100 µL of the extract showed lower intensity of fluorescence, and thus lower intrinsic cellular DNA oxidation compared to control (without S. fruticosa). Conclusions: The results from this study suggest that the water-soluble extract of S. fruticosa leaves protects against both H2O2-induced and intrinsic cellular DNA oxidation in human embryonic kidney 293 cells.

410-415

Authors: Shehla Akbar, Uzma Hanif , Jaffar Ali, Saiqa Ishtiaq

Objective: To establish quality control parameters of a locally occurring medicinal plant, Malva parviflora which is utilized as folk medicine in Sialkot area in Pakistan. Methods: In pharmacognostic studies different types of evaluations were carried out that focus on microscopic, macroscopic, fluorescence analysis and organoleptic evaluations. Results: The distinguishing characters of stem were the presence of parenchyma, cork cells, irregular shape calcium oxalate crystals, simple and compound starch granules and fusiform fi bers with pits. Root microscopic characters were presence of simple and spherical starch granules with rounded or slit hilum, groups of lignified xylem fibers, reticulate vessels, and sieve tissues. Leaves microscopy indicated the presence of paracytic stomata, lignified fibers having pits, spiral and annular vessels, numerous sclereids while in fruit microscopy epicarp, thin walled cells endocarp, thin walled parenchyma and collenchyma of mesocarp and abundant thick walled endospermic cells containing aleurone grains and micro rosette crystals. Macroscopic study of leaves showed, 5-7 lobed reniform-shape, glabrous-surface, reticulate-venation in the leaves. Macroscopic features of roots showed type of root-taproot, surface-glabrous and stem was 1-10 dm tall simple to branched and may be prostrate or ascending. Similarly fruit was of schizocarp type. Conclusions: This study provides the scientific data for the proper identification and establishment of standards for the use of Malva parviflora.

253-261

Authors: Baskar Balakrishnan, Jayappriyan Kothilmozhian Ranishree, Sathish Thadikamala, Prabakaran Panchatcharam

Objective: To identify a potential bacterium which produces antimicrobial peptide (vibriocin), and its purification, characterization and production optimization. The bacteria subjected in the study were isolated from a highly competitive ecological niche of mangrove ecosystem. Methods: The bacterium was characterized by phenotype besides 16S rRNA gene sequence analysis. The antibacterial activity was recognised by using agar well diffusion method. The vibriocin was purified using ammonium sulphate precipitation, butanol extraction, gel filtration chromatography, ion-exchange chromatography and subsequently, by HPLC. Molecular weight of the substance identified in SDS-PAGE. Production optimization performed according to Taguchi’s mathematical model using 6 different nutritional parameters as variables. Results: The objective bacterium was identified as Vibrio parahaemolyticus. The vibriocin showed 18 KDa of molecular mass with mono peptide in nature and highest activity against pathogenic Vibrio harveyi. The peptide act stable in a wide range of pH, temperature, UV radiation, solvents and chemicals utilized. An overall ~20% of vibriocin production was improved, and was noticed that NaCl and agitation speed played a vital role in secretion of vibriocin. Conclusion: The vibriocin identified here would be an effective alternative for chemically synthesized drugs for the management of Vibrio infections in mariculture industry.

267-271

Authors: Khaled Sekkoum, Nasser Belboukhari, Abdelkrim Cheriti

Objective: To investigate the butanol fraction of the water/acetone extract and isolate of the new fl avonoids from Launeae arboescens. Methods: The compounds were isolated by liquid chromatographic methods and their structures were identified by using spectroscopic analysis. Results: The isolated compounds were identified as: 7-O-[α-rhamnopyranosyl 4',5,6-Trihydroxy fl avone 1,4’,5’-Di-Methoxy 7-(5’’-Me Hexan)1-oyl flavanone 2, 3”-isopropyl pyrano [1”:7,4”:6] 3’,4’,5’,5-Tetrahydroxy flavanone 3,5,4’,5’-Tri-Hydroxy 7-(3’’-Me butan) -yl flavanone 4, 5,7-Dihydroxy-2',4',5' –trimethoxy-isoflavanone 5,5,6,7,4'-tetrahydroxy flavonol 6,7-O- [α-rhamnopyranosyl-(1->6)-β-glucopyranosyl]- 4',5,7-tri-hydroxy-flavanone 7,7-O-[α- rhamnopyranosyl-(1->6)-β-glucopyranosyl] 3',5-Dihydroxy 4’-Methoxy flavanone 8. Conclusions: The presence of different types of bioactive flavonoids in Launeae arboescens extract can explain the large ethnopharmacological uses and the potential activity of this medicinal plant.

279-284

Authors: Mst. Mahfuza Khatoon, Mst. Hajera Khatun, Md. Ekramul Islam, Mst. Shahnaj Parvin

Objective: To ascertain analgesic, antibacterial and central nervous system (CNS) depressant activities of ethyl acetate, dichloromethane and carbon tetrachloride fractions of methanol extract of Albizia procera (A. procera) leaves. Methods: Leaves extracts of A. procera were tested for analgesic activity by acetic acid induced and formalin test method in mice. The in vitro antibacterial activity was performed by agar well diffusion method. CNS depressant activity was evaluated by hole cross and open field tests. Results: All the extracts at 200 mg/kg exhibited significant (P<0.01) analgesic activity in acetic acid induced and formalin tests method in mice. Analgesic activity of the ethyl acetate fraction was almost same like as standard drug indomethacin in acetic acid induced method. The CNS depressant activity of the extracts at 500 mg/kg was comparable to the positive control diazepam as determined by hole cross and open field test method. The extracts exhibited moderate antimicrobial activity against all the tested microorganisms (Staphylococcus aureus, Bacillus cereus, Pseudomonas aeruginosa, Esherichia coli, Shigella soneii, Shigella boydii) at concentration of 0.8 mg/disc. The measured diameter of zone of inhibition for the extracts was within the range of 7 to 12 mm which was less than the standard kanamycin (16-24 mm). Conclusions: It is concluded that all the extracts possess potential analgesic and CNS depressants activity. This study also showed that different fractions of methanol extract could be potential sources of new antimicrobial agents.

290-293

Authors: Ramaraj Thirugnanasampandan, Delma David

Objective: To analyse the chemical composition and evaluation of antioxidant, cytotoxic and DNA fr agmentation activities of essential oil of Feronia elephantum Correa. Methods: Chemical composition analysis of hydrodistilled essential oil was determined by gas chromatography-mass spectrometry and in vitro antioxidant activity of oil was determined by DPPH free radical, hydroxyly radical scavenging, metal chelating and prevention of deoxyribose degradation. Cytotoxicity and DNA fragmentation activities against breast cancer cells (MCF-7) were also analyzed. Results: Gas chromatography-mass spectrometry analysis revealed the presence of 24 compounds with caryophyllene oxide (62.29%) as major compound. A considerable antioxidant, cyotoxic and DNA fragmentation activities of oils was observed. Conclusions: The result of this study clearly indicates oil could be useful for food preservation and preparation.

299-305

Authors: Sourov Kuri, Md. Mustahsan Billah, S. M. Masud Rana, Zannatul Naim, Md. Mahmodul Islam, Md. Hasanuzzaman, Md. Ramjan Ali, Rana Banik

Objective: To study the phytochemical and biological properties (antioxidant, anthelmintic and thrombolytic) of methanolic extracts of Enhydra fluctuans Lour., a plant belonging to the Asteraceae family. Methods: The phytochemical evaluation was carried out by qualitative analysis. In vitro antioxidant activity of extract was studied using free radical scavenging assay, ability of reduction, total phenol and total flavonoid contents determination assays. The anthelmintic activity was determined using paralysis and death time of Pheretima posthuma (earthworm) and thrombolytic activity by clot disruption assay. Results: The phytochemical evaluation showed significant presence of flavonoids, triterpenes, carbohydrate, reducing sugars, saponins, phenols, diterpenes, protein and tannin. The antioxidant activity was found significant [IC50=(135.20±0.56) µg/mL] as compared to ascorbic acid [(130.00±0.76) µg/mL]. The reducing power was increased with concentration. Total phenol and total flavonoid contents were (153.08±0.38) mg/mL and (172.04±0.56) mg/mL respectively. The paralysis and death time of earthworms for different concentrations of extract were determined and compared with albendazole. The results showed that 10 mg/mL of the crude extract had similar effect with albendazole. Additionally, the crude extract showed a concentration depended relationship with its anthelmintic property. The clot lysis activity of crude extract was compared to the standard streptokinase’s clot lysis (40.13%) activity and found significant (31%). Conclusions: The study proves that the crude methanolic extract of Enhydra fluctuans Lour. has significant antioxidant, anthelmintic and thrombolytic activity containing wide range of phytochemicals.

312-317

Authors: Nazaire Aïzoun, Rock Aïkpon, Roseric Azondekon, Alex Asidi4, Martin Akogbéto

Objective: To investigate what kind of mosquito sample is necessary for the determination of insecticide susceptibility in malaria vectors. Methods: Larvae and pupae of Anopheles gambiae s.l. (An. gambiae) mosquitoes were collected from the breeding sites in Littoral and Oueme departments. The Centers for Disease Control and Prevention (CDC) susceptibility tests were conducted on unfed male and female mosquitoes aged 2-5 days old. CDC susceptibility tests were also conducted on unfed, blood fed and gravid female mosquitoes aged 2-5 days old. These susceptibility tests were also conducted on unfed and blood fed female mosquitoes aged 2-5 days old and 20 days old. CDC biochemical assay using synergist was also carried out to detect any increase in the activity of enzyme typically involved in insecticide metabolism. Results: Female An. gambiae Ladji and Sekandji populations were more susceptible than the males when they were unfed and aged 2-5 days old. The mortality rates of blood fed female An. gambiae Ladji and Sekandji populations aged 2-5 days old were lower than those obtained when females were unfed. In addition, the mortality rates of gravid female An. gambiae Ladji and Sekandji populations aged 2-5 days old were lower than those obtained when they were unfed. The mortality rate obtained when fe male An. gambiae Sekandji populations were unfed and aged 20 days old was higher than the one obtained when these populations were unfed and aged 2-5 days old. The results obtained after effects of synergist penicillin in beeswax on F1 progeny of An. gambiae Ladji populations resistant to permethrin showed that mono-oxygenases were involved in permethrin resistant F1 progeny from Ladji. Conclusions: The resistance is a hereditary and dynamic phenomenon which can be due to metabolic mechanisms like overproduction of detoxifying enzymes activity. Many factors influence vector susceptibility to insecticide. Among these factors, there are mosquito sex, mosquito age, its physiological status. Therefore, it is useful to respect the World Health Organization criteria in the assessment of insecticide susceptibility tests in malaria vectors. Otherwise, susceptibility testing is conducted using unfed female mosquitoes aged 3-5 days old. Tests should also be carried out at (25±2) °C and (80±10)% relative humidity.

324-328

Authors: Masoud Neghab, Mohammadjafar Momenbella-Fard, Reza Naziaghdam, Narges Salahshour, Maryam Kazemi, Hamzeh Alipour

Objective: To investigate the prevalence of fecundity and other reproductive problems among a group of farmers in Kavar district of Fars province, southern Iran. Methods: A total of 268 randomly selected married male farm workers were investigated. A questionnaire was devised and validated [Cronbach’s α-coefficient (0.81)]. Subjects were directly interviewed and the questionnaire forms were completed for them. Results: The prevalence of current primary infertility among the studied population was about 7.4% (P=0.001). Similarly, 6.3% of farm workers had offsprings with congenital malformations. Finally, 1.5% and 9% of farmers’ wives had a history of stillbirth and abortion, respectively. It was concluded that the prevalence of current primary infertility were higher among farm workers fa milies than in the normal population (P<0.05). Additionally, stillbirth and spontaneous abortion were more common in the wives of farm workers than in the normal population, although the difference did not reach statistical significance. Conclusions: These effects are likely to be attributed to the exposure of farm workers to pesticides.

334-336

Authors: Mohammad Reza Youssefi, Mohammad Taghi Rahimi

The authors report a case of human dermal complications by Ctenocephalides felis felis (cat flea) in a 28 years old woman who dwell in rural area in Mazandaran province, Iran. Furthermore, in the current report clinical manifestations, etiology and different aspects of infestation were discussed elaborately. And the importance of controlling fleas was highlighted as well.