International Conference on Parasitology August 24-26, 2015 Philadelphia, Pennsylvania, USA

Ankita Thakur is pursuing her Ph.D. degree. She is working as Senior Research Fellow (fellowship granted by DST INSPIRE, India) in Department of Zoology, Panjab University, Chandigarh, India. Presently, she has published four papers in reputed journals. Major part of her research is focussed on designing of anti-leishmanial vaccines. As a part of her research she is testing the immunoprophylactic potential of killed parasite antigens as vaccine candidates against murine visceral leishmaniasis.

Visceral leishmaniasis (VL) is a neglected tropical disease caused by protozoan parasites of the Leishmania donovani complex. An estimated 200 000 to 400 000 new cases are reported annually. Control of VL infection till date relies on chemotherapy. The therapeutic armamentarium for VL is currently plagued with several limitations as the available drugs are toxic, expensive and need to be administered for extended periods. So there is an urgent need to find another alternative in the form of a vaccine. Killed vaccines composed of parasite fractions or whole killed parasites reached phase 3 clinical trials, but showed a limited prophylactic efficacy. It seems that major limiting factor for the development of killed vaccines is lack of a suitable adjuvant. Therefore, the aim of the present study was to enhance the protective efficacy of Killed Leishmania donovani (KLD) antigen by using different adjuvants. Inbred BALB/c mice were immunized with KLD antigen alone and in combination with four different adjuvants i.e. alum, saponin, MPL-A and cationic liposomes. Three immunizations were carried out at an interval of two weeks. Two weeks after last booster challenge infection was given and mice were sacrificed on different post challenge days. Protective efficacy of vaccines was analyzed by assessment of parasite burden and generation of cellular and humoral immune responses. All the formulations were found to be immunogenic and imparted significant protection. However, level of protection varied with the type of adjuvant used. Our findings suggest promising antileishmanial potential of KLD as an antigen candidate in combination with cationic liposomes and MPL-A as adjuvants against murine visceral leishmaniasis.

Kgomotso P Sibeko-Matjila has completed her PhD in 2009 from University of Pretoria, South Africa. Subsequently, she was appointed as a Senior Lecturer in the Department of Veterinary Tropical Diseases, University of Pretoria. She has authored and co-authored nine papers in reputed journals and is currently a Primary Supervisor to three PhD and four MSc students.

Theileria parva, an apicomplexan intracellular protozoan parasite, infects and transforms bovine and buffalo lymphocytes causing 2 recognized disease syndromes in cattle, East Coast Fever (ECF) and Corridor Disease (CD). It is still not clear as to why T. parva infections cause different disease syndromes in cattle. In order to study strain-specific changes in gene expression in ECF and CD isolates, Next Generation Sequencing was used to analyze transcriptome of T. parva Muguga and T. parva 7014, respectively. RNA sequence analysis revealed differential expression of 1048 genes between the two isolates. The majority of DEGs were genes encoding sub-telomeric fragment-related protein family (SVSP) (n=85) followed by ribosomal proteins (n=33), membrane/inter-membrane proteins (n=32), signal peptide containing proteins (n=24), transcription/translation/elongation factors (n=23), transporters (n=15) and antigens (n=13). Forty pathways were affected by products of DEGs. The findings of this study provide evidence of variations in gene expression between ECF and CD strains investigated, with most DEGs down-regulated in T. parva 7014 (n=742, 70.8%). Furthermore, genes involved in the ability of the parasite to transform host cells, including members of CD8+ T-cell target antigens, TA9/TP9, SVSP and TashAT gene families, may be the objects of further investigations into understanding the molecular dynamics of ECF and Corridor disease. Many of the DE genes were hypothetical proteins (340), emphasising the need to identify their biological function in order to elucidate their molecular importance in the genetic diversity of T. parva parasites.

Xinping Zhu is a Professor at the Department of Parasitology, School of Basic Medicine, Capital Medical University. She is also the Dean of Department of Mircrobiology and Parasitology.

Trichinella spiralis infection induces protective immunity against re-infection in animal models. Identification of the antigens eliciting acquired immunity during infection is important for vaccine development against Trichinella infection and immunodiagnosis. The T. spiralis adult cDNA library was immunoscreened with sera from pigs experimentally infected with 20,000 infective T. spiralis larvae. More than 40 positive clones were identified; one of the immunodominant proteins was 20 kDa Ts-ES-1 secreted by Trichinella stichocytes and existing in the excretory/secretory (ES) products of T. spiralis adult and muscle larval worms. Ts-ES-1 contains 172 amino acids with a typical signal peptide in the first 20 amino acids. The expression of Ts-ES-1 was detected in both the adult and muscle larval stages at the mRNA and protein expression levels. Mice immunized with recombinant Ts-ES-1 (rTs-ES-1) formulated with ISA50v2 adjuvant exhibited a significant worm reduction in both the adult worm (27%) and muscle larvae burden (42.1%) after a challenge with T. spiralis compared to the adjuvant control group (p<0.01). The rTs-ES-1-induced protection was associated with a high level of specific anti-Ts-ES-1 IgG antibodies and a Th1/Th2 mixed immune response. The newly identified rTs-ES-1 is an immunodominant protein secreted by Trichinella stichocytes during natural infection and enables the induction of partial protective immunity in vaccinated mice against Trichinella infection. Therefore, rTs-ES-1 is a promising candidate for vaccine development against trichinellosis.

Harpreet Kaur is a PhD student in Department of Zoology, Panjab University, Chandigarh (India). She is awarded with UGC-MANF fellowship. She has published 4 papers in reputed journals. She has given oral/poster presentations in different national and international conferences.

Leishmaniasis is a disease that ranges in severity from skin lesions to serious disfigurement and fatal systemic infection. Current treatment is based on chemotherapy which relies on a handful of drugs with serious limitations. Vaccination remains the best hope for control of all forms of the disease. A substantial number of antigens have been identified in past which include gp63, p36/LACK, (CP) B and CPA, HASPB1, LCR1, PSA-2, LeIF, LmSTI1 and TSA, which induce protection against the target parasite; although very few achieve a degree of efficacy likely to make them candidates for single-antigen vaccines. Therefore, multi-antigen/‘cocktail’ vaccines are proposed based on the assumption that such cocktails will show enhanced efficacy. Till date, only one multicomponent vaccine LEISH-F3+GLA-SE has reached clinical trials. Recently studies have been carried out on LD31 and LD51 polypeptides from L. donovani promastigotes which have proven to be potential vaccine candidates. The 36 kDa-glycoprotein is major FML antigenic fraction and designated 'GP36'. The present study is designed to check the protective efficacy of cocktail of low molecular weight antigens. Protective efficacy of different vaccine formulations i.e. 36+51 kDa, 36+51 kDa+ALD, 36+51 kDa+saponin and 36+51kDa+liposome was revealed by significant decline in parasite burden and increased DTH responses. The antibody response was of IgG type with elevated IgG2a and decreased production of IgG1 whereas cytokine levels pointed towards the generation of protective Th1 type of immune response. Among all three vaccine formulations, cocktail of 36+51 kDa+liposome was found to be highly immunogenic and imparted maximum protection.

Aderinola, Adeyinka Aderonke has completed her Master of Science from University of Lagos, Lagos, Nigeria and is currently undertaking her PhD program at Ahmadu Bello University, Zaria, Nigeria. She is a lecturer at the department of Pharmacology, Olabisi Onabanjo University, Ago-iwoye, Nigeria.

Malaria caused by the parasite-P.falciparum is an acute disease which kill an estimated 863,000 people per-year according to WHO report. Malaria chemotherapy failure is beginning to give medical practitioners concern. The continuing spread of multi-drug resistant P.falciparum-malaria now poses a major threat to the tropics. Natural-products are the source of the two most important-drugs currently available to treat severe falciparum-malaria , quinine and artemisinin derivatives, the development of these two important-drugs and utilization of many plants traditionally in various part of the world triggered the search for new, effective antimalarial drugs of natural origin. In this study, Swiss-albino mice ranging from 25-35g were used. The mice were randomly assigned into treatments and controls (negative and positive-control) with five mice per group. Plasmodium-berghei obtained from Nigeria Institute of Medical Research, Lagos were used as donor. Each mouse received 0.1ml of diluted-blood containing 1x 106 P.berghei infected erythrocyte by intraperitoneal-route. Three hours after inoculation of the parasite, the mice in the three treatment groups received the extracts of Newbouldia laevis and cocos nucifera in doses of 200, 400, 600 mg/kg orally for four consecutive-days while the negative and the positive control received normal-saline and 25mg/kg chloroquine-phosphate orally daily for four-consecutive days. On the fifth day, blood sample was collected from tail snip of each mouse, thin smears were prepared, stained with 10%- Giemsa-solution and examined under microscope with an oil immersion objective of 100x magnification power to evaluate percentage suppression. The extract treated mice (200, 400, 600mg/kg) showed decreased parasitemia-level to a highly significant level (p< 0.05).

Yong-long Li was graduated from Tongji Medical College in 1969. He was the director of the Department of Parasitolgy at tongji Medical College. He has published more than 75 papers in reputed journals and has been serving as editorial bord members of journals in China.

Leukemia is a malignancy disorder in leukocytes which occurs in animals as well as human. Current therapeutic for the disease remains unsatisfactory and fatality percentage is higher. Developing new therapeutic strategies for the disease is needed. It has been reported that malaria parasite infection is efficacious to combat some cancers in experimental animals. In the present paper, we studied anti-leukemia activity of malaria parasite Plasmodium yoelii infection in leukemia WEHI-3 cells bearing mice and found that P. yoelii infection significantly inhibited the development of leukemia WEHI-3 cells in the mice and the neoplasm cells infiltrations in the liver and spleen were obviously reduced. Also we demonstrated that malaria infection provided anti-leukemia activity by promotion of immune responses which included increasing percentages cell surface markers of T cell (CD3e) and B cell (CD19), decreasing the amount of the percentage of the cell surface markers of monocytes, (CD11b) and macrophages (Mac-3), inducing the secretion of IFN-r and TNF-α and promotion of natural killer (NK) cell activity.

Jiahui Lei has completed her PhD at the age of 32 years from Tongji Medical College, Huazhong University of Science and Technology. She is Associate Professor in Department of Parasitology. She has published more than 15 papers in reputed journals. And her main research fields are interaction between parasite infection and host and immunological diagnosis of parasite infection.

Cardiac transplantation has been widely accepted as treatment of choice for end-stage heart failure, but organ survival is limited by immune rejection and side effects of traditional immunosuppressants. Therefore, there is an urgent need to develop novel immunosuppressants with few side effects and strong immuosuppression. Several studies show that Toxoplasma gondii infection and administration of soluble tachyzoite antigens (STAg) of T. gondii before transplantation can prolong cardiac allograft survival. However, the effective antigens inducing the protective effects on cardiac allograft are still unclear. In the present study, recombinant surface antigen 1 (rSAG1) , one of the major components in STAg was prepared and injected at 4 d before transplatation. Its effect on cardiac allograft survival was investigated in a mouse model. Moreover, the influence of CD4+CD25+Foxp3+ regulatory T cells (Treg) caused by rSAG1 in the recipient was explored. The results showed that administration of rSAG1 could prolong cardiac graft survival associated with an increased Treg population in splenocytes. This effect coincided with low expression of IFN-γ, IL-4 and IL-17 while increased production of IL-10, TGF-β and IL-12 by splenocytes in the experiment group. Depletion of Treg abrogated the prolonged allograft survival induced by rSAG1 and this effect was associated with decreased expansion of Treg, along with higher levels of IFN-γ, IL-12 and IL-17 and lower levels of IL-4, IL-10 and TGF-β produced by spenocytes. These data suggest that exposure to rSAG1 before cardiac transplantion may induce prolonged allografts survival, which is related to upregulation of CD4+CD25+Foxp3+ regulatory T cells.

Hongwei Zhang has completed his PhD at the age of 39 years from Zhengzhou University and postdoctoral studies from Center for Disease Prevention and Control in USA. He is the director of Institute of Parasite Disease Prevention and Control, Henan Center for Disease Prevention and Control. He has published more than 41 papers in reputed journals and 6 books.

Artemisinin resistance has been confirmed at Cambodia and Thailand. This paper reports the first case of Artemisinin resistance in a 35-year-old Chinese male construction worker with imported falciparum malaria. The patient suffered two days of febrile illness after returning from Nigeria on Oct 1, 2014. The main symptoms were febrile with the highest axillary temperature of 40℃, headache, and chill. A peripheral blood smear showed parasitemia (2000 asexual parasites/μl) with Plasmodium falciparum. The patient had three times medical history of malaria and was administrated quinine, and stopped treatment after he was afebrile in Nigeria during 2009-2014. The patient was administrated oral total 320mg/2.56g Dihydroartemisinin- Piperaquine (40mg/0.32g per tablet) for 2 days and intravenous total 3060mg artesunate for 19 days, which was 60mg artesunate every 6 hours for 3 days(first dosage double), then every 8 hours for 8 days and then each day for 8 days. Axillary temperature of the patient was 37.5-37.8℃ on Day 1-Day 4 and the patient was afebrile on Day 5. Blood microscopy revealed falciparum parasitemia (10000-20000 asexual parasites/μl) during Day 1 to Day 12. Then falciparum parasitemia decreased and was negative on Day 19. The patient was cured and leaved hospital on Day 24. According to WHO classification of antimalarial drug efficacy(Parasitemia on Day 3 with axillary temperature ≥37.5℃) , the patient must be considered an early treatment failure. With increasing number of workers and travelers into malaria endemic areas more attention should be paid in detect the emergence of artemisinin resistance.

Wenqi Liu has completed her PhD from Tongji Medical College. She has published more than 20 papers in reputed journals.

Excretory-secretory products (ESPs) are the majority of worm components which can be directly recognized by host immune system.In our current study, a proteomic approach combined by MS with Two-dimensional gel electrophoresis (2-DE) was used to identify ESPs of S.japonicum. By 2-DE, we identified 166 protein spots from adult ESPs and 138 protein spots from egg ESPs. Of these spots, twelve protein spots were identical in both eggs and adult worms, 24 spots were stage-specifically expressed in adult worms and 16 spots were egg-specifical. In analysis of MALDI-TOF/TOF，the abundant proteins of adult worms revealed a confident constitutent of enzymes， structural proteins，and response elements. Of which, four molecules could be recognized by infective sera of two weeks, and two were reactive with sera of six weeks post infection. An enolase screened from egg ESPs was found to strongly react with early-infected sera, and another putative CAP-Gly-domain protein from egg ESPs was also weakly recognized by sera of early infection. Transposase, abhydrolase domain-containing protein 8 (ABHD-8), and other five hypothetic proteins were not be detected by immunoblotting. In brief, and also notably, we identified total 166 and 138 proteins from ESPs of adult worms and eggs, respectively, which mainly involved in redox balance, protein folding, development and signaling, scavenging, metabolic pathways, and immune response modulation. Several uncharacterized S. japonicum proteins have also been reported in present study. Moreover, five antigens, including confident proteins and putative proteins, have been screened to be potential in early diagnosis for schistosomiasis.

Zhu HM has completed her PhD at the age of 32 years from the Second Military Medical University. She is the professor in Department of Medical Microbiology and Parasitology, and the head of the Joint Diagnostic Lab. on Parasitic Diseases of SMMU.

Patients with fever of unknown origin, were found infection with new human parasites. Apart from fever ,they displayed space occupying focus or spots on lung, liver, Multiple “calcified spots” in kidney, splenomegaly, and systemic lymph node enlargement, etc., which vary between individuals. Some pepole are look normal, and were found abnormalities on physical examinations. Others were sever ill, some of them died. On examing the samples taking from lung or liver punctures, bronchioloalveolar lavage fluid(BLF), faeces, urine,and blood, we found single celled parasites which showing various forms, which caused tissue necrosis. With PCR technique, the amplified fragment proved to be Acanthamoeba sp., having highly similarity(99%) with A. griffini, and A. polyphagus, on sequence alignment. Microscopically, parasites in the samples had morphological diversity. Those with forms inconsistant with Acanthamoeba sp. had not been identified.

Ying Liu has completed her Master's degree at the age of 31 years from Zhengzhou University, She has engaged in malaria prevention and control work for ten years. She has published more than 13 papers in reputed journals.

Plasmodium vivax（P.v） malaria is one of major parasitic diseases with a long history in Henan, China, but little is known about the genotypic polymorphism of P.v in this area. This study aimed to investigate the genetic diversity of circumsporozoite protein (CSP) gene in P.v from Henan, China. Thirty-two P.v isolates from Henan province were characterized by analysing the genetic polymorphism of the CSP gene, all the isolates were found to belong to the VK210 type. The isolates tested displayed variations in the peptide repeat motifs with different size（20-22）or different order, eight different sub-types were observed in the isolates analysed. Compared with the VK210 basic repeat units (GDRADGQPA), more than 75% repeat units have base transition non silence at least one time. seven types of repeat elements were observed in the central repeat units, the change mainly occurred in 3, 4, 6, 7, 9, 14, 21, 22 nucleotides, among which,3,21 often silent while 4,7,9,14,22 often non silent, and the sixth nucleotide can occur base transform both silent and non silent base transform. The nucleotide sequences of Henan isolates were compared with the published sequences, from the genetic distance, isolates from Henan were further with those from Africa and Southeast Asia. The results obtained in the present study indicate that the P. v parasite population is highly diverse in Henan province, and the genotyping protocols used in this study may be useful for differentiating the imported cases from the indigenous ones in vivax malaria.

Zhiyue Lv has completed his PhD at the age of 30 years from Xiangya School of Medicine, Central South University, China and postdoctoral studies from Zhongshan School of Medicine, Sun Yat-sen University in 2007. He had researched as a visiting scholar at School of Medicine, Wayne State University 2012- 2013. Dr. Lv currently is the deputy secretary general of Guangdong Society of Parasitology and vice-chairman of Post-doctoral Fellowship of Guangdong Province, China and he has published more than 20 papers on prevention and control of angiostrongyliasis and schistosomiasis.

The rat lungworm Angiostrongylus cantonensis, a food-borne nematode parasite, is the primary cause of human eosinophilic meningitis, meningoencephalitis and other neurological disorders in many tropical and subtropical regions. Genome-scale investigation of permissive hosts (rat) and non-permissive hosts (mouse and human) for A. cantonensis revealed CCL8, a non-permissive host-specific eosinophil chemotactic chemokine, with an extremely high-level expression in central nerve sysytem (CNS) of A. cantonensis infected mice. Here, we aim to determine the effects of CCL8 on mouse and rat models infected with A. cantonensis. Blocking of CCL8 by injection of anti-CCL8 monoclonal antibodies markedly reduced eosinophil infiltration in peripheral blood and brain of mice infected by A. cantonensis, and the larvae migrated into the pulmonary artery of the infected mice after administration of monoclonal antibodies. In addition, cerebral stereotaxis and microinjection of the infected rats with mouse CCL8 significantly increased eosinophil counts in peripheral blood and cerebrospinal fluid of the treated rats, and obviously inhibited larval migration from the rat brain to lung. Moreover, hamster, another mammal with deletion of CCL8 gene, could served as an permissive host for A. cantonensis. In summary, a deletion of CCL8 contributes to larvae migration from brain to lung and adult development of A. cantonensis in permissive hosts and determines host- A. cantonensis adaption, which demonstrates a novel potential drug target for angiostrongyliasis and enriches a comprehensive understanding of host-parasite coevolution.

Prof. Heng Wang used to work as physician for 12 yeas and did her postdoctoral studies from Naval Medical Research Institute, U.S.A. She is the director of the laboratory of molecular parasitology at Institute of Basic Medical Sciences/Chinese Academy of Medical Sciences, School of Basic Medicine/Peking Union Medical College. She is vise-president of Chinese Academy of Parasitology/ Chinese Academy of Zoology, has published 25 papers in SCI journals and more in Chinese.

Red blood cells (RBCs) targeted by Haemosporina parasites especially the Plasmodium parasites, can shed vesicles to regulate host immune response. In this study, we demonstrated that under the conditions of Plasmodium invasion, normal RBCs (with normal hemoglobin A) could actively export a large number of microparticles (MPs) to infected RBCs. We found that in the infected RBCs, human argonaute 2 (hAgo2) protein complexes with hundreds of human miRNAs (hmiRNAs) inside the parasites, and these hAgo2-miRNA complexes were transferred from uninfected RBCs by MPs. The hAgo-binding miRNAs were immunoprecipitated with anti-hAgo2 antibody and sequenced by high throughput RNA-seq. Among the identified miRNAs, miR-451 and miR-140 are predicted to target the mRNAs of a critical parasite antigen - Plasmodium falciparum erythrocyte membrane protein 1 (PfEMP1). Our data indicate that miR-451 and miR-140 can down-regulate the expression of var genes of malaria by targeting the untranslated regions (UTR) of their mRNAs; depleting hAgo2 leads to accelerated parasite infection in culture; and MPs can protect RBCs from the infection. Our results demonstrate for the first time that normal RBCs are able to export Ago2-miRNA complexes via MPs to infected RBCs, providing evidence that mature RBCs have the innate ability of resisting malaria infection. This study may also help explain why RBC-related genetic mutations tend to exist in malaria endemic areas with a long history of heavy disease burden.

Dr. Wafaa has completed her PhD at the age of 35 years from Cairo University and postdoctoral studies from Theodor Bilharz Research Inst. She is the Associate Prof in Parasitology, a premier science and helath service organization. She has published more than 20 papers in reputed journals.

Cysteine proteases are important virulence factors for parasites. Cathepsin B (Sm CB) proteases are abundant in different stages of Schistosoma and it have been identified to induce a level of host-protective immune responses with amelioration of morbidity. In this study, the parasitological parameters, level of immunoglobulins, cytokines profile and hepatic granuloma were assessed to study the effect of immunization of mice with cathepsin B antigen with or without treatment using anti-helminthic drug (PZQ). Multiple small doses of cathepsin B were intraperitoneally injected into naive mice; the first dose of a 100μg of purified (Sm CB) was followed by two booster doses of 50μg each, at weekly intervals. The experimental design included five groups of 10 mice each; four batches of them were infected with a 100 S. mansoni cercariae as follows: One batch served as infected control, another one was immunized prior to infection, the third was pre-immunized and post-PZQ treated and finally the fourth one was post-treated with PZQ only. The fifth group was mice immunized with cathepsin B only. All groups were sacrificed 8 and 12 weeks post infection. The histopathological and parasitological examinations revealed the highest remarkable increase in the percentage of degenerated ova (12%) within the diminished hepatic granulomas and the most significant decline percentage of the worm burden (46%), tissue egg loads (42.8% and 50% for hepatic and intestinal ova, respectively) were experienced by the infected pre-immunized and post-treated mice. The data collected from this research study might be useful in developing potential vaccine against S. mansoni.