Asian Pacific Journal of Tropical Medicine

14

Authors: Zhao Xu, Wei-zhen Gu, Ji-chu Xue, Wen-ming Zheng, Hong-ge Chen, Su-gai Yin, et al

Objective: Japanese encephalitis virus (JEV) is responsible for one of the most serious epidemics of encephalitis in the world. JEV uses pigs as its main hosts and spreads among vertebrates and humans mediated by Culex mosquitoes. The prevention and control of JEV spread in pigs is one of the most effective measures to protect global public health. Interferon-inducible transmembrane proteins (IFITMs) are small membrane-spanning proteins that were identified as innate antiviral factors against multiple pathogenic viruses, especially enveloped viral pathogens. This study aims to verify whether pig interferon-inducible transmembrane proteins (pIFITMs) inhibit JEV and investigate the related molecular mechanisms of anti-JEV. Methods: Transient expression and RNA interference technology were used to overexpress and silence IFITMs gene. Three different cell lines, PK15, HEK293 and Huh7, were transfected with recombinant pIFITMs-expressing plasmids. The lentiviral vectors harboring RNAi sequences targeting pIFITMs were introduced into PK15 cells. Quantitative real-time PCR was used to determine the antiviral activities of pIFITMs through measuring and analyzing the virus copy number of JEV (SA14-14-2 strain) in the supernant of pIFITMs overexpression or silencing cells 48 hours post transfection. The expression of the related proteins was examined by western blot. The fusion vectors inserted with pig IFITM1-EGFP were constructed and introduced into three different cell lines respectively. Then Laser Co-focus light microscopy was used to observe the subcellular localization. The key active amino acids of pIFITM1 were analyzed by investigating the anti-JEV effect of the cysteine mutants produced with PCR site directed mutagenesistechnology. Results: In three different cell lines, PK15, HEK293 and Huh7, all of three pig IFITM proteins, pIFITM1, pIFITM2, and pIFITM3 could inhibit the replication of JEV whether through transient gene over-expression methods or RNA interference silencing. And that, among three pig IFITMs, pIFITM1 showed the strongest anti-JEV effect. The anti-JEV activity of pIFITM1 manifested at the early entry stage. In PK15, BHK21, and HEK293 cells, before virus infection, pIFITM1 was located in the plasma membrane area, and after infection, transferred to the membranous structures outside the nucleus. The S-palmitoylated cysteines at position 50, 51 and 84 of pIFITM1 had significant effect on virus replication. Conclusions: Pig interferon-inducible transmembrane proteins are restriction factors for JEV infection and have potentials in the prevention of virus spread. Our results provide some new sights into understanding the antiviral activity of pig IFITMs.

16

Authors: Denis Naumov, Dina Gassan, Kseniya Kilimichenko, Evgeniya Afanaseva, Elizaveta Sheludko, Victor Kolosov, et al

Background: TRPM8 is a member of TRP channels family known for its sensitivity to cool temperature, increased osmotic pressure, particulate matter, cigarette smoke and products of oxidative stress. This cation channel is widely expressed in the respiratory tract including airway epithelium and vagal nerve endings where it can mediate inflammatory and secretory responses. Previous studies have reported the increased expression of TRPM8 in the respiratory tract of COPD patients. Objective: To investigate TRPM8 expression in nasal epithelium and induced sputum of asthma patients and to estimate its relation to airway hyperresponsiveness induced by cold air. Methods: The study enrolled 43 subjects with mean age of (39.8±1.76) years including 35 patients with mild-to-moderate asthma and 8 patients with chronic bronchitis. Among the asthma patients 43% were steroid-naïve. Lung function was measured by standard spirometry before and after bronchoprovocation challenge with 3 min isocapnic cold air (-20 °C) hyperventilation. Cold air hyperresponsiveness was diagnosed in case of FEV1 falling by 10% from baseline. TRPM8 expression was measured by indirect flow cytometry in nasal brushings and induced sputum. Nasal epithelium cells were gated after exclusion of dead cells and staining with anti- cytokeratin 19 antibodies. Induced sputum was processed with dithiothreitol, filtered and stained for CD45 and cell viability. Extracellular expression of TRPM8 was detected by staining with primary unconjugated anti-TRPM8 antibodies (Alomone Labs) and secondary antibodies labeled with Alexa Fluor 488 (Abcam). Expression level was calculated as normalized median fluorescence intensity (nMFI) and percent of positively stained cells (%Pos). The data are presented as median, lower and upper quartiles (Me (Q1; Q3)). Results: TRPM8 protein was detected on the epithelial cells and sputum macrophages. The expression levels of TRPM8 were significantly correlated in these cells (nMFI R=0.41, P=0.04; %Pos R=0.53, P=0.008). Expression of TRPM8 was increased both in nasal epithelium and macrophages of steroid-naïve asthma patients as compared to the patients receiving maintenance therapy or patients with chronic bronchitis. nMFI values for macrophages were 2.1 (1.95; 2.79), 1.3 (1.10; 1.91) and 1.2 (1.06; 1.35), respectively (P=0.001). %Pos values for macrophages were 38.5 (13.1; 51.1)%, 11.6 (2.55; 20.1)% and 8.0 (3.20; 9.20)%, respectively (P=0.01). nMFI and %Pos values for the epithelium kept the same trend but the differences were mostly insignificant. In addition, we found a correlation between FEV1 in response to cold air hyperventilation challenge and TRPM8 expression (macrophages %Pos R=-0.43, P=0.02; epithelium %Pos R=- 0.43, P=0.08). Cold airway hyperresponsiveness was accompanied by higher TRPM8 expression on macrophages (21.6 (17.1; 40.7)% vs. 11.6 (7.45; 16.55)%, P=0.03) and nasal epithelium (4.2 (3.30; 5.40)% vs. 2.8 (1.10; 4.30)%, P=0.2). Cold airway hyperresponsiveness was not affected by maintenance therapy in our patients. Conclusions: TRPM8 is overexpressed in steroid-naïve asthma patients as well as in asthma patients with cold airway hyperresponsiveness and may be implicated in asthma pathogenesis. The utility of TRPM8 as a diagnostic biomarker or therapeutic target in chronic obstructive airway diseases is of great interest.

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Authors: Yakuba M Bah, Mary H Hodges, Mustapha Sonnie, Abdulai Conteh, Jusufu Paye, Mohame S Bah, et al

Synergy exists between infectious diseases that target the poor and perpetuate poverty. Common strategies of mass drug administration (MDA) have enabled the control and possible elimination of the five previously neglected tropical diseases (NTDs) to help alleviate poverty. Mapping of five NTDs from 2003-2009 in Sierra Leone found lymphatic filariasis (LF) endemic in all 14 districts, soil transmitted helminthiasis (STH) endemic in all 14 districts with moderate or high prevalence, onchocerciasis endemic in 12 districts and schistosomiasis in 7 (non-coastal) districts, while trachoma was below public health interventio====n significance (<5%). Coordinating with MDA, a national integrated NTD program began in 2008 with funding from United States Agency for International Development and donated or procured medicines. The goal was to eliminate LF and control STH, schistosomiasis and onchocerciasis by year 2020. The program reached full geographic coverage in 2010 with effective programme coverage for all four NTDs each year. By 2016-2018, despite one round of MDA missing due to the Ebola outbreak, LF transmission had been interrupted in 9 districts as shown by LF transmission assessment surveys (TAS), STH and schistosomiasis prevalence had decreased to low or moderate levels in all endemic districts with prevalence of moderate/heavy infection of below 1% and 2% respectively. The burden of onchocerciasis had also decreased significantly, i.e. at baseline the microfilaridermia prevalence (in population of 1 year old and above) using the skin snip biopsy was 53%, while a recent impact assessment using OV16 rapid diagnostic tests showed an antibody prevalence of 2% in children of 5-9 years of age using the LF TAS sampling methodology. Despite the health sectors in Sierra Leone having been severely damaged during the war (1991-2002), with adequate and consistent funding, technical support and drug supplies, Sierra Leone has made remarkable progress towards the control/elimination of four targeted NTDs.

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Authors: Helen Keller International and its contribution to elimination of neglected tropical diseases p. 20 Yao-bi Zhang, Mary Hodges, John U Davis, Volkan Cakir, Xavier Alterescu, Victoria J Quinn

Helen Keller International (HKI) is a non-governmental organization, established in 1915. The mission is to save and improve the sight and lives of the world’s vulnerable by combatting the causes and consequences of blindness, poor health and malnutrition. HKI’s work on neglected tropical diseases (NTDs) dates back to 1950s with a pilot program to eliminate blinding trachoma in Taiwan. HKI’s work on trachoma in Tanzania in 1980s and in Morocco in 1990s contributed to the development of the World Health Organization-endorsed SAFE strategy (Surgery, Antibiotics, Facial cleanliness and Environmental improvement) and the creation of the International Trachoma Initiative dedicated to the elimination of trachoma. HKI is a leader in the control and elimination of onchocerciasis (river blindness) in Africa and played an important role in developing an innovative strategy: community-directed treatment with ivermectin (CDTI) to control onchocerciasis. For its multi-country support to control and eliminate onchocerciasis, HKI was awarded the AGFUND International Prize for Pioneering Development Projects for its “Sustainable Control of Onchocerciasis” program in Africa in 2009. With funding from donors such as USAID, DFID, the END Fund and many others, HKI currently supports 10 African countries to control and eliminate one or more of five major NTDs: lymphatic filariasis (LF), onchocerciasis, schistosomiasis, soil-transmitted helminthiasis (STH) and trachoma which includes leveraging medicines procured or donated from pharmaceutical companies. HKI aims to build the capacity of Ministries of Health to conduct community mobilization, mass drug administration campaigns, morbidity management, monitoring and evaluation. Over the past 10 years, through its integrated NTD programs, HKI has successfully supported Burkina Faso, Cameroon, Guinea, Mali, Niger, Nigeria, and Sierra Leone to complete NTD mapping, initiate MDA, scale up to national coverage, monitor the progress, evaluate the impact and scale down (stopping MDA) once objectives have been met. In 2017 alone, across Africa HKI supported the distribution of over 110 million MDA treatments for NTDs as well as the screening of nearly one million individuals for surgery including follow-up surgery for 36000 affected individuals. By 2018, stopping MDA criteria had been achieved for trachoma nationwide in Burkina Faso, Cameroon and Mali and in most endemic districts in Niger, and for LF nationwide in Mali and nearly in Cameroon and in most endemic districts in Burkina Faso, Niger and Sierra Leone. Prevalence levels for onchocerciasis, schistosomiasis and STH have also been significantly reduced in these countries. Moving forward, HKI will continue to collaborate with partners to support African governments to achieve NTD elimination goals and sustain the gains through mobilizing resources, building the capacity of Ministries of Health and strengthening existing health systems to integrate NTD activities. Joining this Alliance opens the door for HKI to collaborate with more partners on research and elimination of NTDs.

22

Authors: Olaolu Moses Aderinola, A Abubakar, Gabriele Poggensee, Adewale Oyeleke Adeoti, Michael Sunday Oguntoye, Patience Abimbola Folorunso

Objective: Leprosy is a chronic, infectious disease complicated with blindness and loss of fingers/toes in some cases. Though the disease is targeted for eradication in Nigeria, the country had the 6th highest prevalence globally in 2013. This study was conducted to evaluate the leprosy surveillance system in Kwara State, assessing its operations, attributes and determining its usefulness. Method: The 2001 updated guidelines for evaluating public health systems of Centers for Disease Control and Prevention was used. Key informant interviews were done with the State Tuberculosis and Leprosy (TBL) Control Officer, and the State Epidemiologists to assess the importance of the public health, purpose and operation of the surveillance system, resources used to operate it as well as the attributes of the system. Questionnaires were administered to Local Government Area TBL supervisors to assess the attributes of the surveillance system. Surveillance data from 2010 to 2014 was analyzed and data quality determined. Results: Incidence of leprosy over the five year period 2010 – 2014 was 62.0%, 59.0%, 54.0%, 67.0%, and 42.0% respectively, with occurrence of the multi-bacillary type in 83.1% to 90.7% newly detected cases. The system was found to be 51.0% sensitive(most hospitals do not have treatment forms for health workers use, community awareness programmes was done in 53.0% of local government areas in last 3 months, contact tracing was however always done), 90.0% simple, 93.0% acceptable and 95.0% timely. Data system was both paper and electronics, based on collection, collation, analysis and reporting done quarterly in a year. The surveillance system is donor-driven and fully integrated with that of tuberculosis and buruli ulcer. Annual surveillance expenditure was about $2 500. Conclusion: Kwara State Leprosy Surveillance System has a low sensitivity with delayed case detection. Training of health workers on early case detection, provision of treatment forms and more community awareness will improve the surveillance system’s sensitivity.

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Authors: Md Misbahul Ferdous, Effat Un Nesa, Atm Rezaul Karim

All over the world approximately 98 countries faces endemic outbreak of leishmaniases, which is a group of parasitic disease. More than 350 million people reside under the risk of these diseases and new cases per year is about 0.7 to 1.3 million. The culprit and contagious agent Leishmania donovani (L. donovani), which is responsible for leishmaniases are breed mainly in South Asian constituency that harbor’s mainly the visceral leishmaniases (VL) globally. The inter-relationship within main etiological factor (L. donovani & remaining species of leishmaniases) and resultant biological events has been confronted in past years. For this reason there have been accentuated condition of reconsidering the formerly categorization. Rendering to the latest WHO data published in 2017 about leishmaniases death in bangladesh grasped to 988 or 0.13% of total mortality. The age of mortality rate is 0.61 per 100 000 of population, that ranks 7th in whole over the world. This disease can present mainly by three forms: cutaneous, mucocutaneous, and VL. Skin ulcer is the main presentation of cutaneous form, where in the mucocutaneous types, it present with skin ulcer with pyrexia, reduced RBC, hepatomegaly and splenomegaly. Whereas, VL well known as Kala-a-zar, which is focal, huddled inside the households, as well as typically cope the poor people. Also, VL stretches the poverty episode; nevertheless, loss of throughput and highly expensive treatment policy for affected families. Etiological factors for infectivity including residing area within 50 m fom the infected area or home as well as malnutrition and household ailments including splintered mud wall. Though the vector control is essential requirement for disease abolition, but there has been a noticeable lack of innovations from many years back. In addition, there have been many sorts of remarkable improvement in the field of diagnosis of disease as well as therapeutic intervention and vaccination protocol development. But also many kinds of obstacles arise. For exclusion of L. donovani induced leishmaniasis from the certain region of our country become a challenge, and the reality is an effective amendment of existing and new tools will be essential requirements. Both strong political and active community participation would be imperative, also need inter-country cooperation and mutual partnerships. Furthermore, appropriate diagnostic tools, treatment policies including effective demographic surveillance mandatory for ensure of successful outcome to elimination of L. donovani. Considerable success of operational research activities in early detection of disease and complete treatment, IVM, and clinical research achieved from National L. donovani elimination program. Though the activities need to strengthen their policies fro effective surveillance, behavioral change communication (BCC), monitoring, and evaluation as a whole. The main obstacles for the elimination program are inadequate human sources, funds, and logistics. These constrains may overcome by build up good communication and partnership with national and international organizations, donor agencies, and sponsorship institutions. Also operating wisely and diligently with policy makers to warrant this efficious platform is bearable.

26

Authors: Fujie Xu

The major causes of death are not infectious diseases any more, but chronic, noninfectious causes of death are prevalent and increasing not only in affluent countries, but also in lower-income and middle-income countries. RAND corporation scanned the challenges for public health over the next 20 years and summarized that although integration of data will play a significant role, there is no single technology or capability that dominates the field. In the coming decade, advanced therapies, such as genome editing techniques, are expected to deliver cures for a variety of inherited genetic diseases. However, in this era of unprecedented optimism and increasing efforts toward making advanced therapies standard treatment in developed countries, it is also a common reality for those living in lower-income and middle-income countries that the path from vaccine licensure to vaccination has been long and uncertain. For example, pneumococcal vaccines have been shown to be safe and effective in preventing lower respiratory diseases, but the potential of the vaccine has not been fully realized due to lack of widespread use. Through China’s efforts of co-building public health capacity in the coming decades with its partners around developing and developed countries, these spatial and temporal dynamics of public health challenges of infectious, noninfectious, and genetic diseases require an arduous balancing act in setting priorities. Particularly, we public health practitioners should remind ourselves that the greatest global health achievements in the previous century are all population-level interventions. The general public and policy makers should be reminded that the global public health capacity must be adequate to sustain global health achievements and prepare well for future challenges.

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Authors: Li-ying Zhu, Fei-fei Yin, Abulimiti Moming, Bo Wang, Li-juan Gao, Jian-wen Ruan, et al

Severe fever with thrombocytopenia syndrome (SFTS) is an acute infectious disease caused by a tick-borne phlebovirus, which was reported to be continuously epidemic in Japan, Korea and in 23 provinces of China since 2010. Most laboratory-confirmed SFTS cases in China have been recorded in 18 Eastern and Central provinces. A few suspected cases with SFTS-like symptoms were reported to the Chinese Disease Prevention and Control Information System from provinces including Guangxi, Guangdong, Gansu and Xinjiang Uygur Autonomous Region (XJUAR), however diagnosis was not confrimed due to lack of molecular biological and virological evidence. Here we reported the first laboratory-confirmed SFTS case in 2017. A resident of Hainan International Tourism Island (HNITI) was bitten by ticks when traveling in XJUAR and had illness onset after returning to HNITI. RT-PCR detected SFTSV RNA in the patient’s serum samples. Antibodies against SFTSV were detected from the patient and the neutralization from serum samples was evaluated. And the samples of person who had close contact to the patient were also investigated. Moreover, a new SFTSV strain was isolated from the serum sample collected from the patient during acute phase of disease. The viral properties and phylogeny were further characterized. In addition, SFTSV was detected positive in ticks collected from XJUAR in 2017, which suggested that SFTSV was more widely distributed than we recognized. Therefore, this study identified the first SFTS case from XJUAR where confirmed cases have never been reported and demonstrated the substantial risk from SFTSV infection via tick bite there. It is also the first importing SFTS case in HNITI, which showed the significant role of human transport in disease spread and indicated that the recently authorized international tourism island may face more challenges for controlling other importing cases from different areas and countries.

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Authors: Ai-xia Zhai, Ai-mei Li, Wu-qi Song, Wen-ping Kao, Jun Qian, Yu-jun Li, et al

Borna disease virus (BDV) is a non-segmented, single negative stranded RNA virus. It is highly neurotropic, but the infection is not cytopathic to the infected cells. BDV is widely infected in vertebrates, causing abnormal behavior and multiple pathological changes. Horses and sheep are the main natural hosts of BDV. Borna disease (BD) is characterized by progressive mononuclear encephalomyelitis. The BD clinical manifestations include ataxia, depression, rotation movement, mandatory standing, hitting objects with body and paralysis, and they usually last for 1-3 weeks. BDV infection causes a significant health problem in animals and economic loss for the society. However, BDV natural infection frequency in the raised animals in northern China, is poorly understood, but is required for understanding BDV infection epidemiology. BDV infection can be diagnosed through detecting viral proteins, antibodies and nucleic acid. The fluorescence based quantitative PCR is commonly used for BDV diagnosis because of the superior sensitivity. The assays for detecting plasma BDV specific antibodies include immunofluorescence, Western Blot and ELISA. Western Blot offers comparatively high specificity, which can detect different antibodies on the same blot/strip, and is often used as the confirmatory assay. This study investigated the infection prevalence of BDV in the meadow that raised horses and sheep in northern China. Blood samples were collected from a herd of healthy horses and sheep that were meadow raised in Hailin City, Heilongjiang Province, China. The uncoagulated blood samples were used to isolate peripheral lymphocytes and plasma. We selected Mongolian horses which are stocky, with relatively short but strong legs and a large head. The selected Han sheep is relatively small, has short tail and weighs 35-45 kg on average. BDV p40 mRNA was detected by Taqman based quantitative RT-PCR in lymphocytes of horses and sheep. BDV His-phosphoprotein (P) and His-nucleoprotein (N) fusion proteins were isolated with Histidine coated beads and the purity and immunological activity of the isolated viral proteins were assessed and verified by SDS-PAGE and Western Blot. Plasma BDV antibodies in horses and sheep were detected using the purified fusion proteins as antigens. Quantitative PCR assay showed that 50.00% (16/32) of the tested horse samples were positive for BDV p40 mRNA, and 20.83% (5/24) positive for BDV p40 mRNA in the sheep samples. The purified BDV His-P fusion protein in SDS-PAGE electrophoresis migrated at expected 24 kD size. A known specific BDV P antibody verified the immunogenicity of the purified BDV P in Western Blot, and BDV persistently infected human oligodendrocytes as positive control. The purified BDV His-N fusion protein appeared at the expected 40 kD position and the immunogenicity was also verified. The positive rates of BDV P antibody and N antibody were 11.11% (4/36) and 5.56% (2/36) in the tested horses, respectively. The positive rates of BDV P antibody and N antibody were 13.16% (5/38) and 7.89% (3/38) in the tested sheep, respectively. In conclusion, we utilized the expressed BDV His-P and His-N fusion proteins as antigen and established a modified Western Blot assay for detecting plasma BDV antibodies. Our data showed the direct evidence that natural BDV infection occurred in the meadow raised healthy horses and sheep in northern China.

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Authors: Li-cheng Guo, Na Li, Ming-ming Zhao

Objective: Dendrobium officinale (Orchidaceae) (D. officinale) is one of the world’s most endangered plants with great medicinal value. It is mainly distributed in south China, and is often used as auxiliary treatment for a variety of tropical diseases. The strictly demanding for growing environment and climate conditions making its wild resources endangered. In nature, the relationship between seeds of D. officinale and fungi must be established by symbiotic system. Our purpose is to analyze the molecular events involved in the interaction between fungus and plant during this process, and provide scientific basis for the protection of its germplasm resources. Methods: Beta-1,3-glucan (Glu) is a key enzyme in the biosynthesis of polyamines, which is essential to the basic physiological and biochemical processes of plants. In this study, the full-length cDNA of Glu gene was obtained from symbiotic germinating seeds of medicinal plants by rapid amplified cDNA terminal (RACE) - PCR, and its expression characteristics were analyzed for the first time. Results: Glu has a full length cDNA of 1 317 bp, with one open reading frame (ORF). The deduced protein was 356 amino acids (aa) with molecular weight of 37.24 KDa and a theoretical isoelectric point of 5.17. The deduced β-1,3-Dextran protein, without signal peptide, had three transmembrane domain each contained 23,17 and 20 aa. Multiple sequence alignment and phylogenetic analysis showed that the sequence similarity between beta-1,3-glucan and rice beta-1,3-glucan was higher than that of dicotyledons. The expression pattern analyzed by qPCR showed that Glu transcripts were expressed in four tissues (non-infected fungi) without significant change. In symbiotic germinating seeds, the expression level is 23.67 times higher than that of ungerminated seeds. Conclusion: These results indicated that β-1,3-Dextran was possible involved in the symbiotic seed germination of D. officinale, and played an important role in the symbiosis system of mycorrhizal mutual.

34

Authors: Aisha Khan, Hamza Farooq, Sami Simsek, Muhammad Asif Gondal, Haroon Ahmed

Background: Hydatidosis is a zoonotic parasitic disease and a serious human and animal health problem in many endemic areas of the world. It is caused by the tapeworm parasite of Echinococcus granulosus sensu lato. This parasite is endemic in the neighboring countries like China, Iran and India. There are limited studies on CE in Pakistan. Methods: This study primarily aimed to determine the prevalence of hydatidosis in slaughtered livestocks in Chakwal district (Chakwal, Bhaun, Kallar kahar, Choa saidan shah and Balkasar) from June to December 2015. The livestocks were inspected after slaughtering and the ages, breeds and organ wise prevalence were recorded. Results: A total of 6 095 animals (sheep, goat, buffaloes and cattle) of different genders, ages and breeds were inspected after slaughtering. Overall prevalence was 8.55% (88/1 045) in sheep followed by 8.42% (88/1 045), 6.90% (4/58), 2.99% (138/4 608) in cattle, buffaloes and goats, respectively. Liver wise prevalence was 7.75%, 6.90%, 0.06%, 1.30% in cattle, buffaloes, goats and sheep, respectively while lung wise prevalence was 7.55%, 0.67%, 0.00%, 0.00%, in sheep, cattle, buffaloes and goats, respectively. Male (9.50%) were more infected than female (1.96%) in all hosts. Breed wise prevalence was higher in desi breed of cattle followed by crossbreed and other breeds. In goats, teddhi breed was more infected then beetal while in sheep Afghani breed was more infected than the others. Cattle of 6-9 years groups were more infected than 0-3 and 3-6 years age groups. Buffaloes of the only age group of 6-9 years were infected while in goat and sheep 3-6 years were more infected than 0-3 years. According to the statistical analysis there was a significant difference (P<0.05) in prevalence of hydatidosis when ages, genders and organs were compared. Conclusion: The hydatidosis is a serious parasitic problem in the study area for livestocks. The disease should be monitored on regular basis in other areas of Pakistan and future studies should be carried out to explore the real prevalence in Pakistan.

36

Authors: Aisha Khan, Sami Simsek, Haroon Ahmed

Objective: Cystic echinococcosis (CE) is a zoonotic parasitic disease and a neglected infectious disease affecting more than one million people globally. It is caused by the tapeworm parasite of Echinococcus granulosus sensu lato. It is endemic in the neighbours of Pakistan. However, there are limited studies on molecular epidemiology of CE in Pakistan. Methods: Discrimination of Echinococcus species has been done mostly by morphologically and less work has been done by molecular methods in Pakistan. There are limited studies have been conducted to explore the molecular epidemiology of Echinococcus species in human and livestock. In human just retrospective studies have been conducted and there is a serious lack of studies on molecular basis. Results: So far in Pakistan just four studies have been published on genotyping of Echinococcus spp. common sheep strain (G1) and buffalo strain (G3) in livestock (small and large ruminants) and Echinococcus granulosus sensu stricto (G1-G3) in buffaloes, while Echinococcus granulosus sensu stricto (G1-G3), Echinococcus multilocularis, Echinococcus canadensis (G6/7) in human have been reported. Based on these four studies clearly shows that CE is yet neglected in Pakistan. These findings indicate that due to the higher population of sheep and buffaloes, G1 and G3 strains are highly prevalent in human and animals. Association of stray dogs with sheep herds and other livestock at the grazing sites is responsible for the higher prevalence of CE that acts an active role in the transmission of CE while the wild animals and camel are responsible for the less infection. Conclusion: There is dire need of more studies on molecular identification of CE to understand its species diversity and molecular epidemiology in Pakistan. It is an important step toward management of echinococcosis based on One Health Concept.

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Authors: Gao-yao Hu, Feng-Liang , Ji-zhen Wu, Hui Kuang, Zhi-ming Zhang, Wen-fang Long

Objective: With the increasing attention to health influence by ecology environment and climate changes, it is important to explore the characters of drinking water quality and meteorological factors at tropical areas of South China. This study aims to study the water quality of municipal source water and to elucidate the relationship between water quality changes and climatic factors at Haikou. Methods: By analyzing the data of water quality indexes of source water in the past five years from July 2013 to April 2017, the representative indexes as following were analyzed: Fecal coliforms (FC), ammonia nitrogen (NH4+), Nitrate(NO3-), iron(Fe), permanganate Index(CODMn), fluoride(F), arsenic(As), chloride (Cl-). The methods of normal test, rank test and rank correlation were used to explore the water quality and the infuence by precipitation, temperature and sunshine. Results: Only the CODMn was normal distribution for the above indexes. For the five years, Mean of FC was of 110 MPN/L, NH4+ was 0.18 mg/L, Fe was 1.07 mg/L, CODMn was 1.67 mg/L, F was 0.14 mg/L, As was 0.0020 mg/L, NO3- was 1.12 mg/L, Cl- was 7.10mg/L. By the rank correlation test, CODMn, NH4+ and Fe was positive correlated with temperature, it was respectively (r=0.483, P=0.049), (r=0.310, P=0.045), (r=0.423, P=0.039). There is negative correlation for nitrate and chloride between temperature, it is respectively (r=-0.577, P=0.003) (r=-0.317, P=0.042); the indicators are not related to other two climatic factors. Conclusion: The source water has a higher concentration of iron. The temperature is an important factor that affects the local source water quality of Haikou. It is necessary to strengthen removing CODMn, NH4+ and Fe at the torrid period for drinking.

40

Authors: Ru-xuan Zhang, Na Li, Jin Jin, Ting-ting Jiang, Yu-wei Wu, Li-ting Zhou

Objective: To define the effect of PM2.5 exposure on NF- κ B signaling pathway and airway inflammation, and to reveal the role of NF- K B signaling pathway in airway inflammation induced by atmospheric PM2.5 in rats. Air pollution condition plays an important role in inflammatory response process. Previous studies had suggested that atmospheric PM2.5 was a common environmental problem, influencing almost 98% Southeast Asian populations. Therefore, to study the effect of atmospheric PM2.5 on airway inflammation and its mechanism is of great significance. Methods: Atmospheric PM2.5 from urban traffic trunk roads was collected and prepared into the dry powder. Twenty-four wistar rats (half male and half female) were divided into four groups. Rats in low concentration, middle concentration and high concentration groups were given PM2.5 saline suspension of respectively 1.5, 7.5, 37.5 mg/kg by intratracheal instillation, while rats in control group were given saline. HE stain was used to observe the inflammation reaction of the airway tissue. The levels of TNF-α and IL-1β in rats’ bronchoalveolar lavage fluid (BALF) and serum were measured with ELISA. The expression level of IκB-α protein in the airway of rats was detected by western blot. SPSS 17.0 was used for statistical analysis. Results: The organ coefficient of trachea in the group receiving 1.5 mg/kg PM2.5 was significantly higher than that in control group. In group receiving 7.5 mg/kg PM2.5, the lung tissue inflammation was obvious and the structure damage was mild, while in group of 37.5 mg/kg PM2.5, the airway inflammation was mild and the structure damage was severe. In the serum of rats exposed to the 1.5 mg/kg PM2.5, the level of inflammatory factor TNF-a was significantly lower than that in the control group. The level of TNF-a in BALF of rats exposed to PM2.5 was higher than the control group, and with the exposure dose increasing, the level has decreased. The expression of IκB-α protein in lung tissue of rats exposed to the 1.5 mg/kg PM2.5 has increased than in control group. Conclusion: Atmospheric PM2.5 exposure can damage rats’ airway, affect the level of serum inflammatory factor TNF-α, and the protein expression level of IκB-α. With all considered, the NF-κB signaling pathway may play a role in regulating airway inflammation induced by atmospheric PM2.5.

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Authors: Feng Liang, Gao-yao Hu, Ji-zhen Wu, Meng-mi Wei, Ya-li Zheng, Mei-xiu Zhou, et al

Objective: Both antibiotic resistant bacteria (ARBs) and antibiotic resistant genes (ARGs) are considered as one of the most dangerous environmental pollutants in the 21st century by the WHO. This study aims to understand the ARGs of Klebsiella pneumoniae (K. pneumoniae) with multiple drug resistance bacterium from the marine water collected from a beach at Sanya Bay, and to explore the antibiotic resistance mechanism of the K. pneumoniae, providing a basis for exploring the transfer of drug resistance genes of beach, and preventing and controlling the health risk of entertainment population. Methods: The sample of marine water were collected and screened by Mcconkey plate. The drug sensitive test was detected by Merieux VITEK2, The DNA was extracted and one strain of 16srDNA was sequenced and identified as K. pneumoniae. Whole-genome resequencing was performed using Illumina HiseqXten platform, and the obtained sequences were compared with NCBI blasting. The reference bacterium were multi-resistant K. pneumoniae HS11286. Plasmids were extracted and the resistant genes were identified. Results: The ARGs encoding protein was 117/4801 (identity > 40%) and the carrying rate was 2.436 9%. The identity of following ARGs of OKPB, sul1, rpoB, ef-tu, phoP, sul2, AAC(6’)-ib-cr, QnrB, floR, aadA16 were more than 99%. The strain showed resistance to ampicillin, ticacillin/clavulanic acid and chloramphenicol, and was intermediate to ampicillin/sulbactam, compound sulfamethoxazole, ciprofloxacin, minocycline. Preliminary positioning showed that Qnrs, sul1, tetA, cat, QacE were carried on plasmid. Conclusion: The multiple drug resistant bacteria strain has a variety of different resistant phenotypes, some ARGs can be easily spread by plasmid. It probably will bring exposure risk to people for entertainment. Sensibility of some antibiotics were on the brink of resistance, It is necessary to tracking corresponding antibiotics pollution and strengthening monitoring of ARBs and mobile resistant elements of bacteria.

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Authors: Qiong-jun Xu, Li-kang Li, Ri-fei Yang, Jing Zhou

Objective: To understand the epidemiology of Chlamydia trachomatis (C. trachomatis) infection in genital tract in Sanya, and to provide scientific basis for adjusting the existing prevention and control measures. Methods: The cases of C. trachomatis infection in genital tract reported by medical institutions in Sanya from 2012 to 2016 were collected through the disease information management system for descriptive epidemiological analysis. Results: The average annual incidence of C. trachomatis infection in genital tract of Sanya from 2012 to 2016 was 273.02 per 100 000, with an average annual growth rate of 8.40%, and the incidence of C. trachomatis infection in genital tract has been increasing for 5 years (χ2=55.591, P< 0.001). The cumulative number of reported cases of C. trachomatis infection in genital tract in males was 2 617, and that in females was 5 287, the ratio between men and women was 0.49:1. The difference in the incidence rate between men and women was statistically significant(P < 0.05). The cumulative number of reported cases in Tianya District was the highest (7 662 cases), followed by Haitang District (242 cases), and no cases were reported in township hospitals and community health service centers. In the age distribution, the incidence of C. trachomatis infection in genital tract was the highest in the group aged 20-29 years, which was 155.30/100 000, and lowest in the group aged over 60 years, which was 0.97/100 000. The occupation was mainly engaged in houseworkers or unemployed and commercial services. Conclusions: At present, the incidence of C. trachomatis infection in genital tract of Sanya is at a high level. It is necessary to expand screening, strengthen education, and attention should be paid to reduce cross-infection and monitoring drug-resistant strains.

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Authors: Aisha Khan, Sami Simsek, Haroon Ahmed

Cystic echinococcosis (CE) is zoonotic neglected tropical parasitic disease. It is endemic in the some Asian countries like China, Turkey, Iran and India. In Pakistan it is neglected and limited studies have been conducted on epidemiology of CE due to the lack of the fundings. It is not explored yet in Pakistan to estimate incidence and disease burden. In Pakistan recent emergence of CE and its transmission reservoirs are threating to have a higher incidence rate in near future. We are very grateful to Hanian Medical University, China for providing an excellent opportunity under “The Belt and Road Tropical Medical Alliance” which is inspired by the strategy of the “One Belt & One Road Initiative”. This forum will provide direction towards World Health Organization’s call for strengthening tropical disease prevention and research. Taking this opportunity, data on endemicity of CE from Pakistan will be shared in front of scientific community at First International Conference of Tropical Medicine at Hanian Medical University, China. In this way the international cooperation, establishment of research laboratories, collaborative research projects, technology transfer and scientists training will be done under the umbrella of this platform for the screening, control and management of CE in Pakistan. In this international forum of institutions of higher learning, medical institutions, scientific institutions and public health institutions of member countries of “The Belt and Road Tropical Medical Alliance” will not only promote and help the control and management of echinococcosis but also open new directions towards other neglected tropical diseases in Pakistan.

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Authors: M Ferudun Celikmen, Sina Ercan

Buildings that are used as residential areas by humans, mines they work in, vehicles they travel by or snowy mountains they go for sports can turn into deadly traps within seconds due to earthquakes, landslides, tsunamis, floods, bombings, traffic accidents or avalanches. Main survival factors in the post-disaster period are prevention injuries as well as detecting the location of the survivors and the rescued. The reality of the situation of persons who lost their lives in such traps, the severely injured, and the ones who survived must be analyzed. Rational prevention methods against possible crush injuries due to collapsing buildings have been considered in the light of the field and simulation experience we gained and suggestions have been presented to reduce mortality and morbidity. Our work has been conducted with the aid of medicine based on proof, appropriate observation as well as sampling and experimental methods. Developed countries with higher building standards and existence have their action plans to construct buildings with, so that those buildings would not collapse in case of earthquakes and similar disasters. Underdeveloped and developing countries on the other hand, usually adopt these plans exactly the same, without questioning against what and why these should be applied. As there are unlimited possibilities of travelling in todays world, it is not possible to estimate place, country or circumstances under which a person could experience a disaster. Therefore a global approach concerning worst case scenario led by earthquakes has been proposed considering different models of behavior in different countries and societies to increase the chance of survival to a maximum and to reduce injuries to a minimum level.