Asian Pacific Journal of Tropical Disease

596-602

Authors: Walter Chingwaru, Jerneja Vidmar

Objective: To present a comprehensive protocol for the processing of hepatitis E virus (HEV) infected samples and propagation of the virus in primary cell cultures. Methods: Hepatitis E was extracted from porcine liver and faecal samples following standard protocols. The virus was then allowed to attach in the presence of trypsin to primary cells that included porcine and bovine intestinal epithelial cells and macrophages over a period of up to 3 h. The virus was propagated by rotational passaging through the cell cultures. Propagation was confirmed by immunoblotting. Results: We developed a comprehensive protocol to propagate HEV in porcine cell model that includes (i) rotational culturing of the virus between porcine cell types, (ii) pre-incubation of infected cells for 210 min, (iii) use of a semi-complete cell culture medium supplemented with trypsin (0.33 μg/mL) and (iv) the use of simple immunoblot technique to detect the amplified virus based on the open reading frame 2/3. Conclusions: This protocol opens doors towards systematic analysis of the mechanisms that underlie the pathogenesis of HEV in vitro. Using our protocol, one can complete the propagation process within 6 to 9 d.

608-610

Authors: Naeem Bukhari, Afshan Saleem, Abdul Jabbar, Sadiq Noor Khan, Bilal Ahmad, Nasir Habib, Abdul Haseeb, Azhar Khan, Muhammad Zahid Ali, Mohammad Adeeb khan, Aftab Ahmad Khan

Objective: To analyse seropositivity rates of salmonella with seasonal variations in the population of Taxila and the surrounding rural areas. Methods: The study was conducted among 760 suspected patients with symptoms of fever, headache, nausea and decreased white blood cells count screened for the typhoid fever. Blood samples collected from the suspected patients were tested for seropositivity by slide agglutination (Widal test) and Immunochromatographic test, i.e. Typhidot. Results: From overall 760 suspected patients of typhoid fever only 192 (25.26%) samples were positive for Widal and Typhidot test. The peak seropositivity rates were identified during the months of April–June, while decreased cases were observed from January to March. Age wise distribution of typhoid fever reflected that age groups of 10–15 years and 25–35 years were at higher risks of developing enteric fever with respect to age groups of 5–10 years and 61–70 years, respectively. On gender basis evaluation, females (24.49%) were slightly at low risks of developing typhoid fever than males (25.9%). Conclusions: The present study highlights a higher burden of typhoid fever in Taxila and the surrounding areas population that directly reflects the poor hygienic condition and contamination of drinking water. The frequency of typhoid fever fluctuates with seasonal variations as higher rates found during the summer as compared to winter season.

615-621

Authors: Bamidele Akinsanya, Minasu Pentho Kuton

Objective: To study the bioaccumulation of heavy metals from Synodontis clarias (S. clarias) and Chrysichthys nigrodigitatus (C. nigrodigitatus) with their parasitic fauna. Methods: A total of 50 specimens of each fish species (n = 100) were examined. The fishes were subjected to parasitological investigation while 3 g of intestinal tissue of S. clarias and C. nigrodigitatus samples were digested with nitric acid (10 mL). The tissues were then heated until brown fumes disappeared. The samples were allowed to cool and distilled water was added to make up to 50 mL in a standard flask. The filtrate was examined using the atomic absorption spectrometer. The fish hosts were weighed and measured with the aid of digital weighing balance and measuring board, respectively. Results: The Chi-square distribution was significant at 0.01 level (χ2 = 2.16, P < 0.01). A nematode of the family Camallanidae (Procamallanus spp.) was found in S. clarias and a trematode (Siphodera ghanensis) was found in C. nigrodigitatus. The total lengths of C. nigrodigitatus in females and males were (15.05 ± 3.27) cm and (20.35 ± 4.20) cm (P < 0.01), respectively. Also the total lengths of S. clarias in females and males were (21.07 ± 2.80) cm and (20.76 ± 2.37) cm (P < 0.01), respectively. The mean value for the condition factor of both S. clarias and C. nigrodigitatus were more than 2.0. The concentrations of the trace elements in the fishes were Pb > Zn > Mn > Fe > Cd (not detected) and Mn > Zn > Fe > Pb > Cd (not detected), respectively, while in the nematode, Procamallanus spp. and trematode, Siphodera spp. were Pb > Mn > Fe > Zn > Cd and Mn > Fe > Zn > Pb > Cd, respectively. In the water and sediment, the distribution of heavy metals were Fe > Mn > Zn > Pb > Cd and Fe > Mn > Pb > Zn > Cd, respectively. Conclusions: The findings of the concentrations of the trace elements in the aquatic habitat as well as the sediment were below the permissible limit of Federal Ministry of Environment. These findings confirmed that the aquatic habitat was adequate for fishing activity and that the consumption of fish species therein are safe. However, it should be noted that there was bioaccumulation of trace elements in the fish tissues which should not pose any danger to man. Therefore, a regular monitoring of the levels of trace elements in the water body as well as in the fauna should be regularly undertaken.

627-630

Authors: Chukwuebuka Iniobong Ikenna Ugochukwu, Nebolisah Omekam, Emmanuel Ikenna Ugochukwu

Objective: To determine the incidence of Dirofilaria immitis (D. immitis) in dogs using the wet mount and buffy coat techniques for rapid detection of microfilaria in blood samples collected from dogs, to compare the two techniques for quick detection, to find if there is age susceptibility in the incidence of dirofilariasis in dogs presented at the Veterinary Teaching Hospital, University of Nigeria, Nsukka and to find out if there are breed and sex variations in the incidence of dirofilariasis in dogs presented at Veterinary Teaching Hospital, University of Nigeria, Nsukka. Methods: Blood samples were collected from the cephalic vein of 119 dogs. The blood samples were aseptically collected via cephalic venepuncture of each dog, collected into a tube containing ethylene diamine tetraacetic acid as anticoagulant, stored in an ice pack box at 5 °C and processed and examined for microfilaria using wet mount and buffy coat techniques. Results: A total of 4 dogs were positive for D. immitis microfilaria giving a prevalence of 3.36%, more male (4.83%) than female (1.75%) dogs were affected in this study. Although there was no significant difference between both groups, the prevalence was the highest in cross breeds (6.66%), moderate in local breeds (3.63%) and absent in exotic breeds (0.00%). Although there was no significant (P < 0.05) difference amongst the 3 groups, only adult dogs were found positive for D. immitis microfilaria. Conclusions: Based on the results of this present study, both the wet mount and buffy coat techniques can be used at the discretion of the clinician and in the absence of modified Knott’s filter test, ELISA test and other diagnostic imaging techniques, in the rapid detection of microfilaria in blood samples from suspected cases of dirofilariasis.

633-637

Authors: Mona Mohamed Marzouk, Sameh Reda Hussein, Ahmed Elkhateeb, Mai Mohamed Farid, Lamyaa Fawzy Ibrahim, El-Sayed Saleh Abdel- Hameed

Objective: To investigate the phenolic profile of Rorippa palustris (R. palustris) compared to Egyptian Rorippa species to evaluate their chemosystematic significance as well as screen cytotoxicity of R. palustris extract. Methods: The chemical components of defatted hydroalcoholic R. palustris extract were analyzed using liquid chromatography-electrospray ionization mass spectrometry technique. The cytotoxic activity was evaluated by using MTT assay against four carcinoma cell lines. Results: Sixteen compounds corresponding to flavonoids (kaempferol and quercetin derivatives), phenolic acids (gallic, coumaric and ferulic acids derivatives) and an anthocyanin (delphinine derivative) were identified or tentatively characterized, of which ten compounds were detected for the first time from R. palustris. Moreover, R. palustris extract showed a moderate activity against MCF7 and A549 cell lines at 100 mg/mL with cell viability of 47.3% and 65.4%, respectively. Conclusions: The phenolic profile and the morphological characters of R. paluatris seem to be in relation with those of Rorippa indica, which can be considered as an indicative parameter for its medicinal importance.

642-649

Authors: Engwa Azeh Godwill, Marcellus Unaegbu, Aniakor Uchenna Esther, Osuji Amarachukwu Gloria, Agbafor Nwonu Kingsley, Olayinka Ayobami Aiyegoro, Okoh Anthony

Objective: To investigate the antioxidant and antidiabetic activities of the seed and leaf extracts of Chrysophyllum albidum (C. albidum). Methods: After assessing the in vitro ferric reducing power and hydrogen peroxide scavenging activities as well as the flavonoid and flavanol contents, the seed and leaf extracts were administered to diabetic rats for 7 days. The animals were sacrificed and serum was obtained for the determination of blood glucose level while liver sample was used for the quantification of glycogen level as well as lipidic peroxidation and catalase activity. Results: Seed and leaf extracts of C. albidum showed ferric reducing activity and very high hydrogen peroxide scavenging potential. After the administration of treatment in diabetic rats, there was a significant decrease (P < 0.05) in blood sugar level and a significant increase (P < 0.05) in liver glycogen level in Groups 3 and 4 animals administered the leaf and seed extracts respectively compared to Group 1 (the negative control). Also, catalase activity and malondialdehyde levels increased in Groups 3 and 4 administered the extracts compared to Group 1 animals (the negative control). Flavonoids and flavanol were present and significantly higher (P < 0.05) in the leaf than seed extract. In all, the leaf extract showed the greatest activities. Conclusions: These results suggest that the leaf and seed extracts of C. albidum possess both in vitro and in vivo antioxidant activities in scavenging free radicals as well as antidiabetic activity, and as such, a potentially important compound in antidiabetic drug discovery.

654-655

Authors: Udit Chauhan, Neeraj Jain, Sunil Kumar Puri, Lalit Garg, Monika Singh

Bacterial infections of the biliary tree are common infections of the biliary system which frequently lead to life-threatening sepsis. Parasitic infections of the biliary tree like ascariasis are not uncommon. Most adult worms reside into the extrahepatic biliary system. Intrahepatic existence is not commonly described. Urgent recognition of the intrahepatic existence of this common parasite is of paramount importance in order to start timely treatment of this lifethreatening infection. Authors described a case of intrahepatic ascariasis in a young male who was diagnosed radiologically and thereafter managed with endoscopic retrograde cholangiopancreatography and antibiotics.

660-667

Authors: Najeeb Ullah, Abida Parveen, Rahat Bano, Iqra Zulfiqar, Mukharma Maryam, Sadia Jabeen, Amna Liaqat, Sohail Ahmad

Antimicrobial susceptibility testing against pathogenic microorganisms is the most significant task of clinical microbiology laboratory. The present study was therefore designed to review the in vitro and in vivo protocols of antimicrobial bioassays of various medicinal herbal extracts against a diversity of pathogenic microorganisms. Plants have a broad variety of antimicrobial agents which are extensively used as herbal drugs against different microbes. The review covers the antimicrobial techniques and antimicrobial bioassays of medicinal herbal extracts against different bacterial and fungal strains from 2000 onward. Plants have diverse concentrations of bioactive constituents such as alkaloids, saponins, tannins, terpenoids, steroids, carbohydrates, proteins and lipids. These phytochemicals are used against an extensive range of bacteria (Escherichia coli, Mycobacterium, Corynebacterium pervum, Bordetella pertusis, Klebsiella pneumoniae, Salmonella typhi), viruses (simian-virus, retrovirus) and fungi (Pseudomonas aeruginosa, Aspergillus fumigatus, Aspergillus flavus, Fusarium solani). A variety of antibiotics (tetracycline, terramycin, ampicillin) has also been isolated from different medicinal plants. This review was therefore intended to explore the techniques used for antimicrobial activities of herbal medicinal extracts.