Asian Pacific Journal of Tropical Biomedicine

385–388

Authors: Afzan Mat Yusof, Najat Hashim, Muhammad Lokman Md Isa

Objective: To identify the prevalence of Cryptosporidium from goats in three types of farm management systems in Terengganu, Malaysia and to determine the Cryptosporidium species infecting goats by using 18S rRNA. Methods: A total of 478 fecal samples were randomly collected from goats in three farms; 199 samples were collected from intensive farm, 179 samples from semi-intensive farm and 100 samples from extensive farm. The samples were processed by using formolether concentration technique and stained by using modified Ziehl–Neelsen. Positive samples were performed by using nested PCR analysis by using 18S rRNA. Results: Out of 478 goats, 207 (43.3%) were found to be infected with Cryptosporidium. Goats reared under the intensive farm management system reported the highest prevalence of infection (49.7%), followed by intensive farm management system (41%) and the lowest prevalence was reported in the goats reared under semi-intensive management system (37.4%). Conclusions: The identified species found in goat was Cryptosporidium parvum. Future study on the zoonotic transmission of Cryptosporidium parvum in goats needs to be done in order to find the source of transmission of this parasite.

397–400

Authors: Andi Nur Fitriani Abubakar, Suminar Setiati Achmadi, Irma Herawati Suparto

Objective: To determine the structure of triterpenoid isolated from avocado seeds and the cytotoxic effect on MCF-7 and HepG2 cells. Methods: The powder sample was macerated with ethanol, followed with separation of the extract by column chromatography. The target compound was monitored on thin layer chromatography plate and reagent Lieberman–Buchard. The isolated compound was characterized by spectral analysis, mainly ultraviolet, infrared, and liquid chromatographymass spectroscopy and their spectroscopic data with those reported in literature were compared. In vitro cytotoxic activity was investigated against Vero, MCF-7, and HepG2 cell lines using MTT assay. Results: A triterpenoid compound was isolated from ethanol extract. The extracts, fraction (F3), and the isolated compound showed a significant cytotoxic activity against all investigated cell lines. MTT assay showed that the triterpenoid isolate inhibited cell proliferation of MCF-7 and HepG2 cell line with the IC50 values of 62 mg/mL and 12 mg/ mL, respectively, and was safe to normal cells. Conclusions: The results of the present study reveal that triterpenoid from avocado seeds have the potential for further development as anticancer agents.

406–410

Authors: Edy Setiti Wida Utami, Sucipto Hariyanto, Yosephine Sri Wulan Manuhara

Objective: To study asymbiotic seed germination and mass propagation of Dendrobium lasianthera which is one of the endangered medicinal orchids using seeds. Methods: The 14 weeks old hand pollinated seeds were sown on Vacin and Went (VW) solid medium supplemented with various concentrations of peptone (1, 2, 3 g/L) and without peptone which was used as control treatment. At the 4, 8, and 12 weeks after the seeds were sown, seed germination and shoot formation were investigated. To evaluate the role of organic nutrient additives on subsequent shoot development and root formation, particular shoots with about 1 cm length contains 1–2 leaves obtained from the seeds germination was cultured on VW medium additives with different of organic nutrient: 15% coconut water, 2 g/L peptone, 150 g/L banana homogenate, and without organic nutrient was used as control. After 16 weeks of culture, the plantlet height, number of leaves, number of roots, leaf length and root length were recorded. Results: The supplementation of 2 g/L peptone in VW medium was proven to be suitable concentration for seed germination (100%) and shoot formation with 84.0% the protocorm development to phase 5 (shoot). VW medium containing 15% coconut water was effectively improved the shoot development, with well developed roots and leaves compared to the other treatment and 95% of acclimatized plantlets survived. Conclusions: This protocol is an efficient way for the in vitro mass propagation of this Dendrobium lasianthera.

416–420

Authors: Kanchanat Ternchoocheep, Damrassamon Surangkul, Sukhgij Ysothonsreekul

Objective: To investigate the effect of asiatic acid (AA) on the differentiated human neuroblastoma SH-SY5Y cells cytotoxic-induced by cholesterol. Methods: Human neuroblastoma SH-SY5Y cells were either exposed to different concentrations of AA or treated with different doses of cholesterol to reveal their responding viability by MTT assay. The selective 1 mmol/L concentration of AA was then used to test for either the protective or the recovery effects on the cells treated with 250 mmol/L concentration of cholesterol. Results: AA has a propensity to directly increase the viability of differentiated human neuroblastoma SH-SY5Y cells. Cholesterol has significant cytotoxic effect on those cells in a concentration-dependent manner. AA has the ability to slightly recover the viability of the differentiated culture cytotoxic-induced by cholesterol but could not protect those cells from cytotoxic-induced by cholesterol. Conclusions: High concentrations of cholesterol were observed to be harmful to the neurons and AA had a slight effect of reducing neuronal death caused by cholesterol.

428–431

Authors: Wimonrat GunTang, Nathamon Kamonvoradej, Chitchanok Chomchat, Sangrawee Suriyakan2, Sangob Sanit3, Jintana Wongwigkarn, Nophawan Bunchu, Damrongpan Thongwat, Supaporn Lamlertthon

Objective: To investigate the prevalence of Candida spp. and the virulence factors of Candida albicans (C. albicans) isolated from external surfaces of blow flies collected from Mae Sot, Tak Province, Thailand. Methods: The blow flies were collected by sterile sweep nets from three areas in Mae Sot. Yeast isolation was first performed on Sabouraud dextrose agar (SDA) supplemented with chloramphenicol and cycloheximide. The yeast isolates were then identified by using chromogenic agar, a yeast identification test kit, a germ tube formation test and a carbohydrate utilization test. The b-hemolysis was determined on 7% sheep blood agar, while phospholipase activity was measured on SDA agar supplemented with 10% egg yolk suspension. Antifungal susceptibility testing was determined by broth micro-dilution testing against ketoconazole and amphotericin B. Results: The prevalence rate of Candida spp. on the external surfaces of the blow flies was 78.1%. All C. albicans isolated from the blow fly demonstrated b-hemolysin and potent phospholipase activities and 47.1% of C. albicans were resistant to ketoconazole with MIC values 128 mg/mL. Conclusions: The results indicate that blow flies could play an essential role in the transmission of potentially pathogenic and antifungal resistant C. albicans into the environment. Further investigation on other virulence factors and genetic relatedness among isolates from the blow flies, the environment and clinical specimens is required to confirm this role.

437–442

Authors: Guntur Berlian, Olivia Mayasari Tandrasasmita, Raymond Rubianto Tjandrawinata

Objective: To study the regulation of trombinol on thrombopoietin, an essential regulator of thrombocyte production. Methods: Effect of trombinol on thrombopoietin regulation was evaluated at the mRNA and protein levels in human hepatoma HepG2 cells. The mRNA expressions were revealed by PCR and real-time PCR, while the protein expressions were analyzed using western blotting and human ELISA kit. Statistical differences between the test were determined by student's t-test with P < 0.05 was considered statistically significant. Results: Trombinol significantly increased the expression of thrombopoietin at the level of mRNA and protein secretion in HepG2 cell lines. Trombinol with the concentration of 15 mg/mL, positively induces 2.5-fold of thrombopoietin expression. Up-regulation of GABP, a transcription factor of thrombopoietin, is suggested to be involved in cellular regulatory mechanisms of trombinol. Here, our result shows convincing evidence that trombinol affects the thrombopoietin productions in vitro. This molecular explanation of thrombopoietin's stimulating function is in line with the traditional use of Psidium guajava for treatment of diseases involving thrombocytopenia. Conclusions: Thrombopoietin stimulating function of trombinol could be potentially considered as one of alternative treatment for thrombocytopenia-related cases, including post chemotherapy shock, dengue fever and liver failure.

450–454

Authors: Yatnita Parama Cita, Achmad Suhermanto, Ocky Karna Radjasa, Pratiwi Sudharmono

Objective: To explore secondary metabolite of bacteria-associated Xestospongia testudinaria from Tanjung Kasuari, Sorong, Papua. Methods: The antimicrobial activities of extracts against two Gram-positive bacteria (Staphylococcus aureus) and three Gram-negative bacteria (Pseudomonas aeruginosa, Eschericia coli and Salmonella typhi) were determined by disk diffusion dilution method. Results: The test showed that of 15 isolates of symbiont bacteria, 6 isolates were successfully isolated and coded, namely, Xp 4.1, Xp 4.2, Xp 4.3, Xp 4.4, Xp 4.5 and Xp 4.6. Of the six bacterial isolates, isolated Xp 4.2 was found to have more powerful antibacterial activity than any other isolates of symbiont bacteria. Antibacterial activity assay for the n-hexane soluble fractions, ethyl-acetate soluble fractions, and n-buthanol soluble fractions revealed more powerful anti-bacterial activity than any other soluble fractions. Phytochemical screening showed alkaloid and steroid/triterpenoid, while identification for isolate of Xp 4.2 bacterial showed bacteria. Conclusions: Metabolites of bacterial associated with marine sponge Xestospongia testudinaria promise to be developed into antibacterial agents.

460–465

Authors: Enayatollah Seydi, Shabnam Babaei, Amir Fakhri, Jalal Pourahmad

Objective: To explore the cytotoxicity effects of Caspian cobra (Naja oxiana or N. oxiana) venom on hepatocytes and mitochondria obtained from the liver of HCC rats. Methods: In this study, HCC was induced by diethylnitrosamine (DEN), as an initiator, and 2-acetylaminofluorene (2-AAF), as a promoter. Rat liver hepatocytes and mitochondria for evaluation of the selective cytotoxic effect of N. oxiana venom were isolated and mitochondria and cellular parameters related to apoptosis signaling were then determined. Results: Our results showed a raise in mitochondrial reactive oxygen species (ROS) level, swelling in mitochondria, mitochondrial membrane potential (Djm) collapse and release of cytochrome c after exposure of mitochondria only isolated from the HCC group with the crude venom of the N. oxiana (12.5, 25, and 50 mg/mL). This crude venom also induced caspase-3 activation (P < 0.001) in the hepatocytes obtained only from the HCC rat liver. Conclusions: Based on the over all results, we suggested that N. oxiana may be considered as a promising complementary therapeutic agent for the treatment of HCC.

473–477

Authors: Rajiv Ravi, Zary Shariman Yahaya

Objective: To investigate the prevalence, phylogenetics and DNA barcoding of Zeylanicobdella arugamensis (Z. arugamensis) from crimson snapper (Lutjanus erythropterus), Jerejak Island, Penang, Malaysia. Methods: Experiment was conducted with 200 fish specimens of cultured Lutjanus erythropterus from Jerejak Island, Penang, Peninsular Malaysia. The water temperature and length for each fish were measured prior to parasites examination. Next, the morphological identification of parasites was performed. Genomic DNA from parasites was extracted for further molecular analysis. After PCR amplification, phylogenetic tree was constructed. The lowest Bayesian information criterion scores showed that the most compatible model is Tajima and Nei. Finally, data sets of cytochrome oxidase subunit I gene sequence and trace file have been submitted to Barcode of Life Data System. Results: The prevalence rate of Z. arugamensis was recorded to be 11.5%, and the intensity was 1.48. The low intensity was due to the water temperature recorded in this study (32.9–33.2
C). All the individuals of Z. arugamensis recorded in this study showed a close relationship with species that were recorded in NCBI database (Z. arugamensis DQ414344, Aestabdella leiostomi DQ414305, Pterobdella amara DQ414334 and Cystobranchus meyeri DQ414315) but less relationship with Aestabdella abditovesiculata DQ414300. Finally, the DNA sequences submitted to Barcode of Life Data System in accordance to species have already obtained Barcode Index Number as BOLD: ACM3477. Conclusions: This study has provided an overview of sequence divergence at cytochrome oxidase subunit I gene, DNA barcodes and parasite prevalence of Z. arugamensis.

483–491

Authors: I. Made Artika, Chairin Nisa Ma'roef

Emerging viruses are viruses whose occurrence has risen within the past twenty years, or whose presence is likely to increase in the near future. Diseases caused by emerging viruses are a major threat to global public health. In spite of greater awareness of safety and containment procedures, the handling of pathogenic viruses remains a likely source of infection, and mortality, among laboratory workers. There is a steady increase in both the number of laboratories and scientist handling emerging viruses for diagnostics and research. The potential for harm associated to work with these infectious agents can be minimized through the application of sound biosafety concepts and practices. The main factors to the prevention of laboratory-acquired infection are well-trained personnel who are knowledgable and biohazard aware, who are perceptive of the various ways of transmission, and who are professional in safe laboratory practice management. In addition, we should emphasize that appropriate facilities, practices and procedures are to be used by the laboratory workers for the handling of emerging viruses in a safe and secure manner. This review is aimed at providing researchers and laboratory personnel with basic biosafety principles to protect themselves from exposure to emerging viruses while working in the laboratory. This paper focuses on what emerging viruses are, why emerging viruses can cause laboratory-acquired infection, how to assess the risk of working with emerging viruses, and how laboratory-acquired infection can be prevented. Control measures used in the laboratory designed as such that they protect workers from emerging viruses and safeguard the public through the safe disposal of infectious wastes are also addressed.

493–504

Authors: Khaled Mohamed Mohamed Koriem

ultiple sclerosis (MS) is the most famous autoimmune disease attacking the central nervous system. It attacks people from age 20–50 years old and the females' attacks double than males' attacks. MS is an autoimmune disease affecting principally the central nervous system that causes nerve sheath demyelination, followed by axon damage and paralysis. MS symptoms include muscle weakness, weak reflexes, muscle spasm, diffi- culties in movement and unbalance. Many factors may be responsible for MS: microorganism, virus, smoking, stress, environmental toxins, contaminated diet and gout. MS is widely spread in the population in North Europe and this is related to lack of vitamin D due to decrease of sunlight exposure. MS biomarkers include nitric oxide, interleukin-6, nitric oxide synthase, fetuin-A and osteopontin. MS is not a genetic disease (not transferred from parents into next generations) but MS appears when leukocyte antigen system-related genes are changed in human chromosome 6. The physiology of MS patients is controlled by numbers of biological processes such as activation of immuneinflammatory, oxidative and nitrosative stress pathways. MS includes two main steps: (1) myelin sheath destruction and formation of lesions and, (2) inflammation. Four types of MS can be distinguished: relapsing-remitting, primary progressive, secondary progressive and progressive relapsing. Nine treatments have been accepted for relapsingremitting MS type: interferon b-1a, interferon b-1b, mitoxantrone, natalizumab, glatiramer acetate, fingolimod, dimethyl fumarate, teriflunomide, and alemtuzumab. However, the only treatment used is mitoxantrone for progressive MS with many side effects. Complementary treatments are also used in MS treatments such as vitamin D, Yoga, medicinal plants, oxygen therapy, acupuncture and reflexology.