Asian Pacific Journal of Tropical Disease

449-454

Authors: Fatin Khairunnisa Zainutdin, Mohd Aiman Barudin, Mohammed Abdullah Jainul, Muhammad Lokman Md Isa, Afzan Mat Yusof

Objective: To detect the occurrence of Cryptosporidium oocysts and to assess the physicochemical and heavy metal parameters in two main rivers in Kuantan. Methods: Water samples were collected at three sampling points per river (upstream, midstream, downstream) from Kuantan River and Balok River. Samples were filtered using the continuous flow centrifugation machine followed by immunomagnetic separation technique to isolate Cryptosporidium oocysts and stained using fluorescein isothiocyanate staining. Cryptosporidium oocysts were examined using fluorescence microscope. Physical parameters were assessed in-situ using Cyber Scan PCD 650 multi-parameter instrument. Both chemical and heavy metal assessments were done in the laboratory following the American Public Health Association Standard Methods with slight modification. The parameters attained were compared with the Interim National Water Quality Standards (INWQS) which is the standard vital parameters used to evaluate the safety level of surface water in Malaysia. Results: All samples were positive with Cryptosporidium oocysts. Results for physical parameters were within the range of INWQS in Malaysia. For chemical assessment, results for chemical oxygen demand and biological oxygen demand exceeded the INWQS range in most of the sampling points. Of 23 metal elements assessed, only 9 elements were found. Both Kuantan River and Balok River can be classified under Class III river following INWQS which requires extensive treatment. Conclusions: This study hoped to provide new and updated information on the occurrence of Cryptosporidium and its physico-chemical assessment in two main rivers in Kuantan. Future study on molecular identification of Cryptosporidium in rivers needs to be done in order to identify the source of transmission of this waterborne parasite.

458-462

Authors: Gautam Patra, Sonjoy Kumar Borthakur, Seikh Sahanawaz Alam, Subhamoy Ghosh, Hmar Lalliankimi, Hniang Lalrinkima

Objective: To record the prevalence of parasitic fauna of snakes from different parts of Mizoram, India. Methods: Collected fecal samples of different snakes were examined by sedimentation and floatation techniques. Similarly, blood samples were examined for presence of any haemoprotozoa following Giemsa staining technique. Ectoparasites were identified on the basis of morphological keys. Scanning electron microscopy (SEM) was performed for detailed surface structure studies of few parasites. Results: Sixty one percent (40/65) snakes examined were found positive for parasitic infection. The predominant endoparasites included five species of nematodes, one species of cestodes, one species of tissue protozoa, and two species of haemoprotozoa. Ectoparasites recorded were ticks of the genera Aponomma and Amblyomma. The most abundant nematode recorded was Kalicephalus species. Conclusions: The study indicates that parasitic infection of snakes is quite common in this part of India and deserves attention for zoological studies.

467-471

Authors: Hundaol Girma, Getenet Beyene, Zeleke Mekonnen

Objective: To assess the prevalence of intestinal parasites and associated risk factors among food-handlers working at cafeteria of Jimma University Specialized Hospital, Jimma, Ethiopia. Methods: Socio-demographic and associated risk factors data were collected using a pretested structured questionnaire. Stool and finger-nail specimens were screened for intestinal parasites using direct wet mount and formol-ether concentration sedimentation techniques. Data were edited, cleaned, entered and analyzed using statistical package for social science (SPSS) version 20. P ≤ 0.05 was taken as statistically significant. Results: A total of 94 food-handlers working at cafeteria of Jimma University Specialized Hospital were participated in the study. From the total 148 samples (94 stool and 54 fingernails content) examined, 31 (33%) were positive for one or more parasites. Over all eight types of intestinal parasites were identified. The most prevalent parasite identified was Ascaris lumbricoides (16%) followed by Entamoeba histolytica/dispar (4.3%). There was significant association between parasitic infection and food handlers who did not practice hand washing after defecation and before serving food. Conclusions: Relatively high prevalence of intestinal parasites is detected indicating poor hygiene practice of the food-handlers at the study site. The study also identified finger-nail status, hand washing after defecation and before serving food as determinants of intestinal parasitic infection. It is crucial for provision of regular training on strict adherence to good personal hygiene and hygienic food-handling practices as well as regular inspection and medical checkup of food-handlers.

477-481

Authors: Peerawat Chintrakulchai, Sirawit Vuttichai, Anuwat Wiratsudakul

Objective: To estimate the risk of brucellosis diffusion through goat movement network across Nonthaburi Province, Thailand. Methods: A questionnaire-based survey was performed to collect movement data. A static weighted directed network was constructed and a spatial network distribution was plotted. Alternatively, a stochastic quantitative risk estimation was also calculated. Results: A total of 114 goat farmers responded to the questionnaires. Twenty-two provinces across Thailand were identified in the network. The highest weighted-in- and weighted-outdegree centralities were observed in Bangkok and Kanchanaburi Province at 264 and 68, respectively. Moreover, the highest import risk of caprine brucellosis into Nonthaburi Province was found in Nakhon Pathom at 13.40 ± 1.93 whereas the highest export risk was noticed in Bangkok at 17.45 ± 1.05. Conclusions: The identified provinces should be closely monitored especially in terms of animal movements. This study may help relevant authorities prioritize the high-risk areas and optimize control measures. Our methods could also be applied to other diseases or animal species in different geographical locations.

486-490

Authors: Mubbashir Hussain, Shahzad Munir, Madiha Fatima, Kashif Rahim, Irfan Ahmed5, Abdul Basit6, Mir Zulqarnain Talpur, Nawaz Haider Bashir, Arslan Jamil, Muhammad Ameen Jamal

Objective: To evaluate the prevalence of extended spectrum beta-lactamases (ESBL) in clinical isolates from burn patients using phenotypic and genotypic analyses. Methods: During 2015–2016, a total of 126 samples were collected at a tertiary care hospital, Islamabad. Antibiotic sensitivity and ESBL prevalence were evaluated according to the Clinical Laboratory and Standards Institute, and molecular analysis of the CTX-M type ESBL gene was performed in 225 bacterial isolates from these samples. Results: The most prevalent bacterial species were Escherichia coli (28.4%), Pseudomonas aeruginosa (22.2%), Staphylococcus aureus (19.6%), Klebsiella pneumoniae (16.4%), and coagulase-negative staphylococci (13.3%). Of the 225 bacterial isolates, 89 (39.5%) were found to be ESBL producers. The isolates were highly susceptible to meropenem (88%) and imipenem (84%), followed by the aminoglycoside amikacin (81%). Molecular epidemiology of the ESBL isolates indicated 19% prevalence of CTX-M. Resistance to antibiotics was exhibited by 28% isolates. Conclusions: In the present study, bacteria such as P. aeruginosa, K. pneumoniae, S. aureus, and E. coli isolated from burn patients exhibited resistance to one or more antibiotics and produced large amounts of ESBL. Further studies are needed to investigate the virulence and epidemiology of CTX-M type ESBL in clinical isolates from burn patients.

496-501

Authors: Rahmah Waty, Apon Zaenal Mustopa, Suharsono, Ratih Asmana Ningrum, Hidayah Murtiyaningsih

Objective: To express HBcAg protein (hepatitis B virus subgenotype B3) in Escherichia coli in soluble form. Methods: HBcAg sequence of hepatitis B virus subgenotype B3 was cloned into plasmid pET32a and introduced to E. coli BL21 (DE3). The E. coli was grown in Luria-Bertani (LB) medium supplemented with ampicillin with agitation. Protein expression was induced by adding isopropyl-β-D-thiogalactopyranoside (IPTG) at concentrations of 0.1 mmol/L, 0.3 mmol/L, and 0.5 mmol/L at room temperature (28 °C). The bacteria were dissolved in lysis buffer and lysed by freeze-thawing method then sonication. The fusion protein [thioredoxin A-(His)6tag-HBcAg] was purified using immobilized metal affinity chromatography. The protein expression was analyzed by SDS-PAGE, dot blot, and western blot. Results: This research showed that DNA sequence of HBcAg could be propagated in pET32a and soluble protein was successfully expressed in E. coli. Induction with 0.3 mmol/L IPTG and 4-hour incubation was the best condition to express the HBcAg protein. SDS-PAGE and dot blot analysis showed that HBcAg protein could be expressed in E. coli. Western blot analysis showed that molecular weight of HBcAg fusion protein was about 38.5 kDa. Conclusions: This study confirmed that HBcAg protein could be expressed in soluble form in E. coli.