Day 1 :
Keynote Forum
Raymond P. Goodrich
Colorado State University, USA
Keynote: Prevention of Transfusion-Transmitted Parasitic Infections: Preventing Transfusion-Transmission of Malaria in Africa
Time : 09:30-10:10
Biography:
Dr. Goodrich received his BS in Chemistry from The Ohio State University and a PhD in Chemistry from the California Institute of Technology. As the Executive Director of the Infectious Disease Research Center (IDRC) at Colorado State University, Dr. Goodrich has responsibility for oversight of the Biopharmaceutical Manufacturing and Academic Resource Center (BioMARC), Regional Biocontainment Laboratory (RBL) and the Research Innovation Center (RIC). He also holds a faculty appointment as Professor in the Department of Microbiology, Immunology and Pathology at Colorado State University and an Adjunct Professor of Chemistry at The Ohio State University. Dr. Goodrich has worked in medical research for over 29 years during which he has managed research staff and development programs in the fields of transfusion and transplantation medicine and pathogen reduction technologies. He has been awarded over 58 patents covering technology in these areas and has co-authored over 200 peer reviewed articles and abstracts.
Abstract:
Transfusion-transmitted parasitic diseases remain a major obstacle to the provision of safe blood products for support of patients in many parts of the world. Transfusion-transmitted malaria in parts of Africa provides an example of the magnitude of this issue. Studies have demonstrated that 14-28% of the patients presenting for transfusion in Ghana are susceptible to transmission of malaria through the blood products that they receive. Screening of blood donations to prevent transmission is not feasible, as this could eliminate up to 50% of the donated blood products in a region where blood availability and supply is an issue. Recently developed methods for treating blood products prior to transfusion to inactivate pathogens that may be present have been implemented in various regions of the world. In vitro testing has demonstrated the efficacy of these processes for a number of bacterial, viral and parasitic agents. Until recently, however, the processes have been limited to treatment of platelet and plasma blood products. In regions of sub-Saharan Africa and other parts of the world, whole blood that has not been separated into components represents the major transfused product, which is given primarily for treatment of conditions such as post-partum bleeding, traumatic injury or support of patients with sickle cell disease. The ability to prevent transmission of disease in this setting would require treatment of all components of blood and preferably whole blood in order to keep the process simple and remain cost-effective. This presentation will focus on results from a clinical study on the prevention of transfusion transmitted malaria in Ghana and report the results of a clinical investigation of this approach and subsequent implementation of this method in large scale in the region. In addition, applicability to other disease transmission events for bacterial, viral and parasitic agents will be discussed.
Keynote Forum
David W. Wright
Vanderbilt University, USA
Keynote: New Approaches to Low-Resource Diagnostics
Time : 10:10-10:50
Biography:
David Wright (b. 1965) attended Tulane University in New Orleans, Louisiana where he received a B.S. in Chemistry and a B.A. in Classical Languages and Ancient History. He pursued his graduate work at the Massachusetts Institute of Technology in Cambridge, MA under the supervision of Prof. William. H. Orme-Johnson, focused on understanding the structure and function of the FeMo-cofactor of the enzyme nitrogenase. David joined the faculty of Vanderbilt University in 2001. His research focus spans areas ranging from heme detoxification in hemophagous parasitic infectious diseases to developing tools and diagnostics for low-resource settings to understanding the biomineralization of novel materials in biological systems.
Abstract:
There are over 2 billion people on the planet who live on less than $2.00 a day. What happens when they get sick? How can they afford to go to the doctor and find out why they don’t fell well? The solutions for the diagnostic needs of the people living at the bottom of the healthcare pyramid can be found in our children's toy box, the kitchen counter, or on the poster advertising this weekend’s rave. The creation of ASSURED diagnostic devices requires new, non-traditional approaches for the development of effective tests for parasitic diseases. In this talk, we will highlight how the principles of everyday objects and phenomena can be adapted for highly sensitive and specific medical tests.
This talk will demonstrate how three different everyday objects have been successfully adopted as low resource diagnostics. The concepts originate from origami, the ancient art of Japanese paper folding, coffee rings, like those left on the kitchen counter, and glow sticks, a favorite children’s' toy. These new devices provide new solutions to the challenges of field deployable diagnostic tests for the detection of asymptomatic malaria.
Keynote Forum
Manuel Gonzalez-Ronquillo
Universidad de Zaragoza, Mexico
Keynote: Evaluation of anthelmintic efficacy of a homeopathic product (parafil) and ivermectin on sheep
Biography:
M. Gonzalez Ronquillo, Doctor in Veterinary Medicine at the Universidad de Zaragoza, Spain. FAO Voluntary in AGA division, Rome Italy, and Fulbright in the US Dairy Forage Research, I worked with natural extracts as antihelmintics properties, ruminant nutrition and Curve lactation models in small ruminants.
Abstract:
The incidence of parasites that affects severely the health and productivity of small ruminants has been increased in the last years to due climatic change and misuse of anthelmintic therapies. The aim of this study were assessed the anthelmintic efficacy of an allopathic (Ivermectin, IV) and other homeopathic product (parafil, PF) in naturally infested ewes. To achieve these objectives the fecal egg count reduction test and percentage of larval development for detecting resistance were evaluate on 36 ewes allocated in each one of the three treatments, over a 56 days period. Fecal samples were collected rectally from all animals weekly (0, 14, 21, 28 and 56 days). Mixed model was used to evaluate the effect of anthelmintic product on body condition score (BCS), body weight (BW), average daily gain (ADG), fecal egg count (FEC), efficacy and resistance (PROCMIXED, SAS). The results are shown in the table 1. Parafil showed higher efficacy, ADG and Best FAMACHA score and lower resistance in comparison with IV and control group, therefore is a viable option to reduce gastrointestinal parasites in sheep without compromise the animal health and environmental equilibrium.
- General Parasitology
Medical Parasitology
Veterinary Parasitology
Tropical Parasitology
Vector-borne Diseases
Parasitic Treatment and Control
Location: Prague, Czech Republic
Session Introduction
Francoise Debierre-Grockiego
University Tours, France
Title: Nanoparticle-based vaccine platforms against apicomplexan
Biography:
Françoise Debierre-Grockiego is teacher at the University of Tours (France) in the fields of parasitology and vaccine. She has more than 15 years experience in the study of the biological effects of glycosylphosphatidylinositols from protozoan parasites (Toxoplasma gondii, Plasmodium falciparum, Trypanosoma cruzi). Her knowledge is built on vaccine strategy developed by the team Antiparasitic Biopharmaceuticals experienced in vaccinating animals via nasal route and challenging animals with Toxoplasma or Neospora to evaluate vaccine efficacy against chronic and congenital infection, based on study of humoral and cellular immune responses.
Abstract:
Vaccines based on live attenuated parasites are associated with safety concerns related to reversion to pathogenic form or recombination with wild strains, are expensive, require a cold storage for delivery, are difficult to manufacture and have a short shelf life. Subunit vaccines are more stable and safer but they have lower efficiency. Optimal progress towards novel vaccines against intracellular apicomplexan parasites will depend on several factors such as selection of appropriate parasite antigens and optimal presentation of these antigens to the immune system, including identification of the best adjuvant formulation and route of vaccine administration. Due to the complexity of apicomplexan parasites, vaccine strategies must associate large panel of antigens and adjuvant able to better stimulate innate and adaptive immune responses than in natural infection. Toll-Like Receptors are expressed on numerous cells of the immune system and TLR agonists are promising adjuvants also in farm animals. Mucosal administration of vaccines, by mimicking natural route of infection, is an important approach to induce appropriate protective immune responses to microbial antigens in systemic sites and peripheral blood as well as in most mucosal environments.
Our team has demonstrated that maltodextrin/phospholipid nanoparticles sustain release of all soluble components of Toxoplasma gondii after nasal administration. This formulation led to protection against congenital toxoplasmosis in mice (70% reduction in parasite load) and chronic toxoplasmosis in ewes (no parasite detected) correlated with strong and long-lasting humoral and Th1/Th17 cellular responses (Dimier-Poisson et al. 2015, Ducournau et al. 2017, and unpublished data). We now apply this vaccine strategy against an emerging cause of abortion and infertility problems in cattle, neosporosis due to Neospora caninum. TLR ligands are added as adjuvant to reach sterilizing vaccine.
Heejeong Youn
Seoul National University, South Korea
Title: Summary of Evaluation of Anti-Protozoal Efficacy from Medicinal Herbs Extracts against Eimeria tenella, Toxoplasma gondii and Neospora caninum
Biography:
Hee-Jeong Youn is professor of Department of Veterinary Pathology, College of Veterinary Medicine, Seoul National University, Seoul, 151-742, South Korea. Major interesting research areas are the development of anti-protozoal drugs and the parasites for wild rodents and so on.
Abstract:
Eimeria tenella, Toxoplasma gondii and Neospora caninum are the very important coccidial protozoa in domestic animals and poultry. In order to develop the anti-coccidial drugs, the extracts of 15 medicinal herbs were screeningly evaluated the anti-coccidial efficacies against E. tenella. Sophora flavescens (S. flavescens), Pulsatilla koreana, Sinomenium acutum, Ulmus macrocarpa and Torillis japonica (T. japonica) were the more effective anti-coccidial activities than the other herbs against E. tenella. These five herbs were re-evaluated the anti-protozoal activities against Toxoplasma gondii (T. gondii) and Neospora caninum (N. caninum). S. flavescens and T. japonica were the more effective anti-protozoal activities than the other herbs against T. gondii and N. caninum.
S. flavescens and T. japonica were evaluated anti-protozoal efficacy of medicinal herb extracts in cell culture. These herbs extracts were fractionated by use of high performance liquid chromatography (HPLC). Nine HPLC fractions from herb extracts of S. flavescens and T. japonica were evaluated the anti-neosporal efficacy against T. gondii and N. caninum, and we found 6 fractions (4 fractions of S. flavescens, 2 fractions of T. japonica) which showed good anti-neosporal efficacy in this study.
In the second test, to characterize the chemical components associated with anti-neosporal activity, specific fractions were isolated by HPLC. Gas chromatography mass spectrometry (GC/MS) was then performed to certify the components and to determine their amount in the herb extracts, and these fractions were evaluated in vitro against N. caninum. To determine the ability of each fraction to inhibit parasite proliferation, 3H-uracil incorporation was used to determine parasite replication. Four fracions (3 fractions of S. flavesens, 1 fraction of T. japonica) showed high anti-protozoal efficacy for N. caninum. The constituents of the herb extracts (T. japonica and S. flavescens) were Sophoridane(2), Matridin-15-one(CAS, 1), Furosardonin A(1), Tetraisopropylidene -cyclobutanee, 5,17,beta-Dihydroxy-de-A-Estra-5,7,9,14-Tetraene (1), Furanodiene (1), 9,12-Octadecadienoic acid (Z,Z)-(CAS, 1).
We tested these herbal HPLC fractions in mice, Korean native goats and dogs to determine anti-neosporal efficacy. In this study, we infected Korean isolate of Neospora (KBA-2) to mice, pregnant goats and suckling dogs and we evaluated anti-neosporal efficacy of HPLC fractions of herb extracts (S. flavescens, T. japonica). The mouse which was treated with high dose of S. flavescens (7.125ng/㎖) and T. japonica (7.125ng/㎖) showed improved survival rate to 25% and 41.7%, respectively and showed decreased brain lesion score to 1.81 and 2.23, respectively. In this experiment, the extracts of S. flavesens and T. japonica showed good efficacy in protection of Neospora, and improved survival rates in mice. We could find out that S. flavesens and T. japonica could delay abortion days in pregnant goats but it could not prohibit Neospora infection or treat Neosporosis.
Artem Rogovsky
Texas A&M University, USA
Title: The distribution and diversity of Lyme disease pathogens in the Ixodes ticks in Ukraine
Biography:
Dr. Artem Rogovskyy is a faculty at the Department of Veterinary Pathobiology, College of Veterinary Medicine & Biomedical Sciences, Texas A&M University, USA. Dr. Rogovskyy came to Texas A&M University following extensive research training at the master’s level at Louisiana State University, followed by mentored research at the University of Georgia, and then PhD from Washington State University. In addition to his teaching commitment, Dr. Rogovskyy runs his research laboratory and serves as an Associate Director of the Clinical Microbiology Laboratory in the Veterinary Medical Teaching Hospital. Dr. Rogovskyy is American College of Veterinary Microbiologists-board certified microbiologist.
Abstract:
Lyme borreliosis (LB) is caused by tick-borne spirochetes of Borrelia burgdorferi sensu lato complex (Bb) transmitted by Ixodes ricinus ticks in Europe. As in the United States, LB is of great public health concern in Europe with the Bb prevalence in Ixodes ticks ranging from 6.2% in Poland, 9.4% in Belarus, 11% in Lithuania to up to 40.7% in Russia. LB is currently the most prevalent vector-borne illness in Ukraine. During the period of 2000-2010, 4,597 cases were reported. The objective of the present study was to investigate the prevalence and diversity of Bb pathogens in I. ricinus ticks collected in Kyiv and Chernobyl cities of Ukraine. It was hypothesized that a potential difference in Bb prevalence and diversity of Bb species may exist in the ticks collected from the two different urban ecosystems. As a result of nuclear meltdown, the uninhabited Chernobyl region has been directly exposed to ionizing radiation for over 30 years. On the other hand, Kyiv, the most densely populated megapolis of Ukraine was exposed to radiation at a much lower degree. Questing I. ricinus ticks were collected via flagging from eight recreational areas in Kyiv and within the 10- and 30-kilometer Chernobyl Exclusion Zones (CEZ). A total of 587 and 372 I. ricinus ticks were collected in Kyiv and CEZ, respectively. In our study, collected ticks were subjected to real-time and nested PCR analysis. Resulting amplicons were sequenced to determine Bb genospecies. Our preliminary results reveal that 11.5% and 22.5% of I. ricinus ticks in Kyiv and CEZ, respectively, were positive for Bb DNA. B. afzelii, B. burgdorferi sensu stricto, and B. garinii were the three genospecies detected in ticks. Overall, the present study provides the most current data on the diversity and prevalence of Bb spirochetes in the two cities of Ukraine.
Ikuo Igarashi Obihiro
University of Agriculture and Veterinary Medicine, Japan
Title: Efficacy of combination chemotherapy with clofazimine and diminazene aceturate against Babesia and Theileria infections
Biography:
Dr. Ikuo Igarashi is currently a Professor at the National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Japan and an OIE expert for equine piroplasmosis and bovine babsiosis. He has been done researches on the in vitro cultivation of Babesia and Theileria and its application for drug screening. In addition, he is investigating the molecular mechanisms of invasion and multiplication of Babesia parasites in red blood cells and the development of highly specific and sensitive serological and molecular diagnostic assays. He has published more than 250 parasitology papers in peer-reviewed scientific journals, book chapters, and patents. He has been also serving as a member of the Editorial Advisory Board for Veterinary Parasitology since 2005 and an Academic Editor of PLoS ONE from 2012 to 2015. He is currently a Co-Editor-in-Chief for Veterinary Parasitology.
Abstract:
In the present study, we evaluated the growth-inhibitory effects of clofazimine, currently used for treating leprosy, against Babesia bovis, B. bigemina, B. caballi, and Theileria equi in vitro culture, and B. microti in mice. The IC50 values of clofazimine against the in vitro growth of B. bovis, B. bigemina, B. caballi, and T. equi were 4.5, 3, 4.3, and 0.29 μM, respectively. In mice infected with B. microti, treatment with oral administration of 20 mg/kg clofazimine resulted in a significant lower peak parasitemia (5.3%) as compared to a control group (45.9%), which was comparable to subcutaneous administration of 25 mg/kg diminazene aceturate. However, the growth of parasites was observed in mice after blood transfusions from clofazimine-treated mice on day 40 post-infection when parasites were not found in the blood smears. These results suggest that clofazimine has excellent inhibitory effects against Babesia and Theileria in vitro and in vivo, but it could not completely eliminate parasites in the host. Therefore, we evaluated the combination treatment with clofazimine and diminazene aceturate against piroplasms both in vitro and in vivo for the development of a novel chemotherapy with high efficacy and safety against animal piroplasmosis. The clofazimine-diminazene aceturate combination showed to have additive or synergistic effects on in vitro growths of Babesia bovis, B. bigemina, B. caballi, and Theileria equi. The lower dosages of clofazimine-diminazene aceturate combination showed to have a higher chemotherapeutic efficacy against B. microti in mice as compared to clofazimine or diminazene aceturate monotherapy. B. microti was not detected in blood, brain, spleen, and heart DNA samples by PCR from combination therapy on day 51 post infection. Furthermore, the parasite did not grow in blood-transfused mice from combination therapy groups. All results suggest that the clofazimine-diminazene aceturate combination chemotherapy will be a better choice to treat animal piroplasmosis instead of diminazene aceturate monochemotherapy.
Bogumiła Skotarczak
Szczecin University, Poland
Title: The potential role of Ixodes ricinus ticks in life cycle of Toxoplasma gondii
Biography:
I have been the head of Department of Genetics (University of Szczecin, Poland) since 1998. The main scope of my studies concerns molecular biology of pathogens transferred by ticks (Borrelia burgdorferi sensu lato, Babesia sp., Anaplasma sp., Rickettsia sp., Toxoplasma gondii). I have lectures and seminars for students of Biology, Biotechnology and Microbiology. I have been the member of Editorial Board of Annals of Agricultural and Environmental Medicine, member of honour of Associazione Nazionale di Parasstologia Medica (Italy) since 1995, member of International Society of Evolutionary Protistology (Rockefeller University, New York, USA) since 1999, member of Polish Genetic Society since and member of Polish Parasitological Society since 1978. I am the author of one monograph and author or coauthor of many (more than 120) publications (Annals of Agricultural and Environmental Medicine, European Journal of Wildlife Research, Journal of Parasitology, Scandinavian Journal of Infectious Diseases; Infections, genetics and evolution; Acta Veterinaria Brno, Experimental and Applied Acarology, Ticks and Tick Borne Diseases, Folia Biologica).
Abstract:
Ixodes ricinus as a blood-sucking tick uses many of wild and domestic animal species as hosts. Almost each of Toxoplasma gondii intermediate host may serve as a source of blood for each tick stage. In this way, an infected tick may take part in the spread of T. gondii in the environment through the transmission of this parasite between different host species. In this way, ticks may increase the number of infected specimens among hosts populations.
How Ixodes ricinus takes part in the life cycle of T. gondii ?
I. ricinus takes part in the life cycle of T. gondii as a mechanical or even biological vector. Even though the tick does not transmit T. gondii to other host mammals, it may take infection from them after molting and increases populations of infected ticks. The mammalian hosts digest infected ticks and in this way T. gondii is transmitted, what explains herbivorous infection, rodents and birds infection. In this case, ticks do not transmit T. gondii to other hosts but serve as reservoir host for T. gondii, however it requires more studies.
Ixodes ricinus may help to explain the wide spread of this parasite
Toxoplasmosis is globally distributed, water- and food borne zoonosis caused by that single protozoan and probably one-third of the world’s human population is infected with this pathogen. Human as other of the intermediate hosts may become infected in the main route of infection; it is the ingestion of parasite oocysts in contaminated water or soil and undercooked meat. However, the mechanism which this parasite uses to so large spread is not fully understood, because oral transmission does not explain the common event of this parasite in a variety of hosts. Thus, ticks may help to explain the wide spread of this parasite in almost all geographical areas and warm blooded animals, but there is need to establish whether they are an important infection source for toxoplasmosis.
Our study
We examined 1737 feeding I. ricinus ticks collected from 49 Shetland ponies (Imno Stud, Poland) and 371 questing ticks using B1 gene based PCR and sequencing of positive samples for genotyping of T. gondii. T. gondii genotype I pathogenic for humans was detected in 4.5% of all I. ricinus, including in 2.99% of feeding ticks and in 10.24% of questing ticks and the difference was statistically significant. Our results confirmed the presence of T. gondii in I. ricinus and showed a new geographical habitat of T. gondii occurring in I. ricinus ticks in Poland.
Manuel Gonzalez-Ronquillo
Universidad de Zaragoza, Mexico
Title: The use of calcium soaps with natural extracts (Allium sativum or Salix babylonica) as an anthelmintic in goats
Biography:
M. Gonzalez Ronquillo, Doctor in Veterinary Medicine at the Universidad de Zaragoza, Spain. FAO Voluntary in AGA division, Rome Italy, and Fulbright in the US Dairy Forage Research, I worked with natural extracts as antihelmintics properties, ruminant nutrition and Curve lactation models in small ruminants.
Abstract:
Parasites infections of commercially livestock farms are responsible for economic loses. To combat these effects, most farms administer anthelmintics to their animals at frequent intervals, and without consideration to principles of sustainable integrated parasite management (SIPM). On another hand the use of Garlic (Allium sativum) and Salix babylonica is well know spice with also anti-microbial and anti-parasitical properties. The objectives of this research were to characterize levels of anthelmintic resistance on goats’ and to compare the fecal egg count reduction test (FECRT) and percentage of larval development for detecting resistance. To achieve these objectives, nine goats were allocated in each one of the three treatments, the FECR efficacy was used to evaluate resistance status to calcium soaps (CS) performed with Salix babylonica extract (CSalix 5 % diet), CS performed with garlic (CGarlic 5 % diet) and control (Ctrl) as a natural desparasitant over a 28 days period. Faecal samples were collected rectally from all animals weekly (0, 14, 21 and 28 days). Mixed model was used to evaluate the effect of anthelmintic product on body weight (BW), average daily gain (ADG), BW change (BWC), faecal egg count (FEC), and efficacy (PROCMIXED, SAS). Results (Table 1) of the FEC indicated a reduction (P<0.003) and efficiency (P<0.001) to CSalix on 67 % and CGarlic 66 % compared with the control treatment. The use of calcium soaps in the diet adding Garlic or Salix babylonica is a viable option to reduce gastrointestinal parasites in goats without compromise the animal health and environmental equilibrium.
Kálmán Imre
Banat's University of Agricultural Sciences and Veterinary Medicine, Romania
Title: Comparative detection of Giardia and Cryptosporidium spp. in different water types: microscopy vs. conventional PCR
Biography:
Kálmán IMRE has the most important scientific achievements in the study of the genus Cryptosporidium. His research activity covers a wide variety of epidemiologic surveys and diagnostic methods including microscopic, immunologic and molecular tools. Mainly, the investigations were focused to environmental biology of Cryptosporidium species, molecular epidemiology, diagnosis and treatment of cryptosporidiosis in farm animals and humans from western Romania, with important contributions to the understandings of the complex puzzle of this zoonotic disease.
Abstract:
Statement of the Problem: Giardia and Cryptosporidium spp. are considered two important waterborne parasites. Their detection in the aquatic environment can be achieved by different methods, each one exhibiting advantages and disadvantages. The aim of the present investigation was to compare the Giardia cysts and Cryptosporidium oocysts water detection efficiency of the immunofluorescence assay (IFA) and the conventional polymerase chain reaction (PCR), in order to bring contribution in the developing and/or improvement of effective identification methods in monitoring of water hygiene for both pathogens. Methodology & Theoretical Orientation: One hundred forty different types of water samples (sewage, river, brook, lake, pond, collector channel) were collected through microfiber filtration, concentrated according to USEPA 1623 protocol and examined for the detection of Giardia and Cryptosporidium spp. through immunofluorescence microscopy and PCR techniques. Findings: Giardia cysts were microscopically detected in 48 (34.3%) samples, while the parasite DNA was recorded in 37 (26.4%) samples. Seven (5.0%) samples showed microscopically positive results for Cryptosporidium oocysts, and the protozoa DNA was isolated from 5 (3.8%) samples. Some of the microscopically positive samples showed negative PCR results and vice-versa, such that the Cohen’s kappa index values, used to measure the degree of agreement between methods, were calculated as being 0.514 for Giardia and 0.826 for Cryptosporidium. Conclusion & Significance: The used methods can be considered effective diagnostic techniques in water detection of Giardia and Cryptosporidium (oo)cysts, and their complementary usage offer a promising tool in the monitoring of aquatic environment pollution for both pathogens.
Acknowledgements: This work was supported by a grant from the Romanian National Authority for Scientific Research and Innovation, CNCS – UEFISCDI, project number PN-II-RU-TE-2014-4-1300.