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Comparative analysis of current diagnostic PCR assays in detecting pathogenic Leptospira isolates from environmental samples

May-Ling YAP ; Zamberi SEKAWI ; Hui-Yee CHEE ; Vasantha NEELA ; Han Alan ONG

Asian Pacific Journal of Tropical Medicine.2019;12(10):472-478. doi:10.4103/1995-7645.269908

Objective: To compare the efficiency of routine diagnostic PCR assays in detecting pathogenic Leptospira isolated from water and soils. Methods: Seven routine assays targeting six genes (lipL32, flaB, gyrB, lfb1, secY and ligB) were evaluated and compared on the cultures of two groups of pathogenic Leptospira from different sources. One group included 19 described reference strains recovered from infected human or animals, and another group included 22 environmental isolates from recreational and residential sites in Malaysia. The latter have been confirmed for presence of pathogenic Leptospira DNA. PCR positivity or detection sensitivity of each assay was determined and compared between the two groups. Results: Validation on reference strains showed 100.0% PCR sensitivity for all assays except ligB-PCR (95.0%) that failed to amplify Leptospira interrogans serovar Pomona. In marked contrast, there was a notable decline in sensitivity in the environmental isolates (lipL32-PCR, 95.5%;flaB-PCR, 90.9%; gyrB-PCR, 77.3%; lfb1-PCR, 59.1%; secY-PCRs, 40.9% G1/G2-PCR, 36.4%; ligB-PCR, 13.6%), implying a large genetic distance between the two groups, as well as nucleotide polymorphism among environmental isolates. Conclusions: High proportion of false-negative PCR results suggests a need of prudent selection of primers in detecting environmental pathogenic Leptospira. These findings offer valuable insights on the extensive biodiversity of genus Leptospira and its impact on the efficacy and development of molecular detection tool.

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Ranjit SAH ; Shusila KHADKA ; Meenu CHAUDHARY ; Rafael TOLEDO ; Lucrecia ACOSTA

Asian Pacific Journal of Tropical Medicine.2019;12(10):479-482. doi:10.4103/1995-7645.269909

Rationale: Microbial keratitis caused by coinfection with more than one species of pathogens is a severe condition with an unfavorable prognosis. Patient concerns: An immunocompetent Nepali woman complained of pain in the left eye, redness, watering and decreased vision for 5 months. Interventions: The patient was discarded and accurately diagnosed with coinfection with Fusarium sp. and Acanthamoeba sp. The habit of washing the eyes with tap water from a domestic storage tank was the most likely source of infection since it was found to be contaminated with cysts of Acanthamoeba sp. The woman received eye drops of fluconazole and natamycin (5%), cefazoline (50 mg/mL), atropine, and tablets of itraconazole (100 mg), which were later switched to eye drops of clotrimazole (1%), natamycin (5%) and voriconazole (1%), and tablets of itraconazole. A full thickness penetrating keratoplasty was performed followed by treatment with eye drops of voriconazole (1%), natamet (5%), ofloxacin, atropine and carboxymethylcellulose for one week. Outcomes: After treatment, the condition of the patient significantly improved and was discharged one week after keratoplasty. Lessons: This is the first report of Acanthamoeba keratitis in Nepal and the first report of coinfection with Fusarium in this country and highlights the importance of early diagnosis of microbial keratitis both in single microorganism infections and coinfections, even in no contact lens wearers.

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Ebola virus disease: Recent advancs in diagnostics and therapeutics

Supriya JAGGA ; Ashish Ranjan SHARMA ; Chiranjib CHAKRABORTY ; Sang-Soo LEE ; Chiranjib CHAKRABORTY

Asian Pacific Journal of Tropical Medicine.2019;12(9):385-395. doi:10.4103/1995-7645.267581

Ebola virus disease (EVD) is associated with haemorrhagic fever in humans and nonhuman primates, with a high rate of fatality (up to 90%). Some outbreaks in human history have proven the lethality of EVD. The recent epidemic of 2014 and 2015 in West Africa was the deadliest of all time (11 284 deaths). To understand the transmission dynamics, we have reviewed the epidemiology of EVD to date. The absence of any licensed vaccines or approved drugs against Ebola virus (EBOV) further highlights the severity and crisis level of EVD. Some organizations (public and private) are making considerable efforts to develop novel therapeutic approaches or vaccines to contain the outbreak of EBOV shortly. Here, we summarized the various potential drugs and vaccines (undergoing multiple phases of clinical trials) that have arisen as an alternative against EBOV, and we highlighted the numerous issues and limitations hindering this process. Alternatively, an increasing focus on strengthening the medical and civic health structure could provide speedy benefits in containing the spread of EVD, as well as offer a resilient foundation for the deployment of novel drugs and vaccines to the affected countries, once such drugs and vaccines become available.

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Long-term safety follow-up of children from a randomized - Controlled phase II b proof - Of - concept efficacy study of the live, attenuated, tetravalent dengue vaccine (CYD - TDV) in Thailand

Kriengsak LIMKITTIKUL ; Weerawan HATTASINGH ; Arunee SABCHAREON ; Danaya CHANSINGHAKUL ; Wut DULYACHAI ; Carina FRAGO ; T. Anh WARTEL ; Alain BOUCKENOOGHE ; Edith LANGEVIN ; Sophia GAILHARDOU

Asian Pacific Journal of Tropical Medicine.2019;12(9):396-403. doi:10.4103/1995-7645.267582

Objective: To investigate the long-term safety of a tetravalent dengue vaccine (CYD-TDV) in children in a phase Π b follow-up study in Thailand. Methods: In the phase Π b study, children aged 4-11 years were randomized (2:1) to receive three injections of CYD-TDV or serve as control at 6-month intervals, with 25 months' active follow-up (active phase). This study was an additional four-year passive surveillance for hospitalized virologically-confirmed dengue (VCD; hospital phase). Cases of hospitalized VCD, severe hospitalized VCD, vaccine-related serious adverse events, and deaths were reported for the total population, with post-hoc analyses by enrollment age (<9 and years). Results: Of 3 997 participants receiving injection, 80.1% were recruited to the hospital phase [2 131 (CYD-TDV); 1 072 (control)]. Eighty-five hospitalized VCD cases were reported in the CYD-TDV group and 46 in the control group during the four-year hospital phase [relative risk (RR): 0.93, 95% confidence interval (CI): 0.64-1.36]. The RR over six years of follow-up was 0.77 (95% CI: 0.57-1.05). In those aged ≥9 years, the cumulative RRs in the active phase, hospital phase, and entire six years were 0.28 (95% CI: 0.08-0.81), 0.51 (95% CI: 0.25-1.05), and 0.42 (95% CI: 0.24-0.75), respectively. In the overall population, there were ten severe hospitalized VCD cases in the CYD-TDV group and five in the control group over six years (RR: 1.00, 95% CI: 0.31-3.75). Conclusions: Over six years of follow-up, in children aged ≥9 years, CYD-TDV administration is associated with a reduced risk of hospitalized VCD.

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Association between measles antibodies in vaccinated and naturally infected mothers with protective antibodies and the occurrence of measles in their children: A cross-sectional study in the Bavi district of Hanoi

Cam Nhat NGUYEN ; Than Huu DAO ; Quynh Ngoc NGUYEN ; Le Thi Quynh MAI

Asian Pacific Journal of Tropical Medicine.2019;12(9):404-408. doi:10.4103/1995-7645.267583

Objective: To determine the concentration and rate of decay of maternal IgG antibodies against measles prevalence in infants of vaccinated or naturally infected mothers and study initial measles immunization occurs in nine-month-old children. Methods: In total, 401 pregnant women and the same number of their subsequent newborns were recruited in the Bavi district of Hanoi in 2016-2017; they were divided into two groups: Older women (born before 1985, n=201) and younger women (born after 1990, n=200). Samples were collected at five time-points; week 36 of pregnancy, birth (cord), and 3, 6, and 9 months after birth. Measles-specific IgG antibody levels were recorded. Results: In total, 77.06% of the 401 pregnant women were seropositive for measles-specific IgG antibodies. A significantly greater proportion of mothers aged 30 and older (88.06%) and their newborn (93.53%) were seropositive compared to the mothers aged 25 and younger (66.00%), and their newborn (72.00%) (P<0.001). The infants of older mothers had significantly higher geometric mean titres (GMT) of measles IgG antibodies than the infants of younger mothers (P<0.001) at all time-points of the study period. The proportion of measles IgG antibodies together with GMT decreased from 82.97% (506.96) at the age of three months to 23.19% (45.22) at the age of nine months. Conclusions: This study provides a profile of maternal antibodies against measles in Vietnamese infants and investigates the early susceptibility to measles in both the infants of vaccinated mothers and mothers with naturally acquired immunity. These data suggest that determining the appropriate age for measles vaccination is paramount for the elimination of measles in Vietnam.

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Chemoprotective activity of aqueous leaf extract of Acalypha wilkesiana against cyclophosphamide-induced toxicity in rats

Chinedu P. ANOKWURU ; Chinedu P. ANOKWURU ; Isaiah Di RAMAITE ; God'Swill N. ANYASOR ; Olutayo S. SHOKUNBI ; Babajide SOPEKAN ; Oyindamola K. OSINUGA ; Olusola E. AFOLABI ; Olamide B. AROJOJOYE ; Chibundu N. EZEKIEL

Asian Pacific Journal of Tropical Medicine.2019;12(9):409-415. doi:10.4103/1995-7645.267584

Objective: To investigate the protective effect of aqueous leaf extract of Acalypha (A.) wilkesiana Muell. Arg (Euphorbiaceae) against cyclophosphamide-induced toxicity in albino rats. Methods: Twenty male albino rats were randomly divided into five groups of four animals each. The control group (I) was fed with pellets and distilled water, while group II was orally administered with only 20 mg/kg cyclophosphamide. Groups III, IV and V were co- administered with 20 mg/kg body weight cyclophosphamide and 110, 220 and 440 mg/kg body weight A. wilkesiana leaf extract, respectively, for 7 d. After treatment, liver and kidney function biomarkers, haematological parameters, liver antioxidants, and mitochondrial membrane permeability transition pore opening were investigated. Results: A. wilkesiana leaf extract significantly reduced (P<0.05) cyclophosphamide-induced increase in plasma aspartate aminotransferase, alanine aminotransferase, creatinine, uric acid and urea. It increased superoxide dismutase, catalase, glutathione-S-transferase activities and reduced glutathione levels. It also increased packed cell volume count, hemoglobin concentration and white blood cell count while inhibiting the induction of mitochondrial swelling. Conclusions: This study demonstrates that aqueous extract of A. wilkesiana leaf protected tissues against cyclophosphamide-induced oxidative damage.

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Curcuma angustifolia ameliorates carbon tetrachloride-induced hepatotoxicity in HepG2 cells and Swiss albino rats

Sudipta JENA ; Asit RAY ; Ambika SAHOO ; Subhashree SINGH ; Noohi NASIM ; Sanghamitra NAYAK ; Diptirani RATH ; Durga Madhab KAR

Asian Pacific Journal of Tropical Medicine.2019;12(9):416-424. doi:10.4103/1995-7645.267585

Objective: To determine the antioxidant and hepatoprotective potential of methanol extract of rhizome of Curcuma angustifolia (MECA) against carbon tetrachloride (CCl

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Inhibitory effects of methanolic Olea europaea and acetonic Acacia laeta on growth of Babesia and Theileria

Amany Magdy BESHBISHY ; Gaber EL-SABER BATIHA ; Naoaki YOKOYAMA ; Ikuo IGARASHI ; Gaber EL-SABER BATIHA ; Oluyomi Stephen ADEYEMI

Asian Pacific Journal of Tropical Medicine.2019;12(9):425-434. doi:10.4103/1995-7645.267586

Objective: To evaluate the antipiroplasmic activities of methanolic extract of Olea europaea (MOE) and acetonic extract of Acacia laeta (AAL) against Babesia and Theileria parasites in vitro and evaluate the chemotherapeutic effects of these extracts against Babesia (B.) microti in vivo. Methods: Fluorescence assay using SYBR Green 1 nucleic acid stain was used to detect inhibitory effects of the two extracts as well as the combination effects of the two extracts with diminazene aceturate and atovaquone on four Babesia species and Theileria equi in vitro while for in vivo experiments, 8-weekold female BALB/c mice were injected intraperitoneally with 1 × 10

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Pharmacological and analytical aspects of artemisinin for malaria: Advances and challenges

Sravani KARRI ; Sanjay SHARMA ; Ginpreet KAUR

Asian Pacific Journal of Tropical Medicine.2019;12(8):339-346. doi:10.4103/1995-7645.262562

Malaria remains a major tropical health burden owing to the development of resistance and decreased sensitivity to the frequently used conventional antimalarial drugs. The drug like artemisinin possesses potent antimalarial activities, but has some limitations. Therefore, new strategies are to be implemented for optimal utilization of artemisinin to improve its therapeutic effectiveness and to overcome its limitations. The present review focuses on present scenario of malaria and pharmacological as well as analytical aspects of artemisinin. Data from 2000 to 2018 were collected from NCBI for understanding the various analytical techniques used for estimation of artemisinin. This review will reveal the facts about artemisinin which can be utilized to develop novel drug delivery system either in a combination or as alone for the wellbeing of the patients suffering from malaria.

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The cholera epidemic of 2004 in Douala, Cameroon: A lesson learned

Christoph HEMMER ; Stefan FINKBEINER ; Emil REISINGER ; Jürgen NÖSKE ; Günter KUNDT

Asian Pacific Journal of Tropical Medicine.2019;12(8):347-352. doi:10.4103/1995-7645.262563

Objective: To describe the outbreak of 2004 with a view of retrospectively identifying factors that might explain the low case fatality rate. Methods: Outbreak data from 4 915 Cholera patients from registers of the Regional Health Delegation in Douala were analyzed using SPSS. Chi-square test, univariate and multivariate analysis were applied. Results: The outbreak started January 2004, peaking at 187 cases per week in February. After a decrease in April, case numbers rose to 688 cases per week in June. The outbreak was over in September 2004 ( <10 cases per week). The case fatality rate was higher in treatment centers with fewer than one nurse per two patients, than in those with more nursing staff. A temporary staff reduction after the first wave of the epidemic was associated with the increase of the case fatality rate during the second wave. This increase was reversed after re-instating full staff capacity. Conclusions: Providing sufficient nursing staff helps to lower the case fatality rate of cholera. Besides a lack of staff, age above 40 years is a risk factor for death in this disease.

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