& Nie P. (2010). 13 We suggest a new analytical framework to predict high-risk snail habitats based on ecological models. None of the parasites and snails species used in this study are endangered or protected species. and snails such as Lymnaea spp., Pomacea canaliculata, and Melanoides spp. https://doi.org/10.1371/journal.pone.0224617, Editor: Angela Monica Ionica, University of Agricultural Sciences and Veterinary Medicine Cluj-Napoca, Life Science Institute, ROMANIA, Received: May 29, 2019; Accepted: October 17, 2019; Published: November 20, 2019. (2018) who observed an unusually high prevalence of intestinal protozoan infections together with schistosomiasis japonica in individuals in Northern Samar, the Philippines contributing to the significant public health and socio-economic burden affecting this population. Having successfully established colonies of Oncomelania quadrasi in aquaria in Manila, the authors tested the effects of various chemicals on them. Based on qPCR results, all aquaria except the negative set-ups were positive to O. hupensis quadrasi DNA, while 7 out of the 9 aquaria had detectable S. japonicum DNA (Table 2). In one part of this study, snails were collected from a site which was previously recorded with very low to zero infection rates. Detection of cercarial DNA of S. japonicum from water samples was first reported as a promising technique to apply in schistosomiasis-endemic areas in China [1416]; however, no trials or field tests have yet been done in the Philippines. In 1940, the first reliable results were and therefore play an important role in the parasites life cycle. Control, prevention and elimination of the disease call for continuous monitoring of previously identified snail sites and searching for new potential snail breeding sites. With malacological surveys currently serving as the conventional method to determine snail colonies and their infection rates, correct identification of O. hupensis quadrasi is paramount to ensure accuracy and reliability of data. No, Is the Subject Area "Snails" applicable to this article? Overall, this method can complement malacological surveys for monitoring of schistosomes in endemic areas of the Philippines, especially those with high risk of human infection. Improving environmental sanitation by building facilities such as toilets and latrines and the provision of safe sources of water for domestic use can greatly reduce environmental contamination and exposure among high risk communities. Yang, Guo-Jing1; Utzinger, Jrg; Sun, Le-Ping; Hong, Qing-Biao; Vounatsou, Penelope; Tanner, Marcel; Zhou, Xiao-Nong 2007. Schistosomiasis (or snail fever) is a disease common among residents (such as farmers, fisherfolk and their families) of communities where there they are exposed to infected bodies of water (such as irrigation systems, rice paddies, swamps). On the right (B) the PCR using S. japonicum primers were done with templates of S. japonicum gDNA (lane Sj), and the non-target gDNA of Fasciola spp. Graduate School, University of the East Ramon Magsaysay Memorial Medical Center, Quezon City, Philippines, Roles [1] It appears likely that there has been certain degree of isolation for these mountainous populations.[1]. "Development of Larval. Schistosomiasis is a snail-borne neglected tropical disease affecting 78 countries and territories in Africa, Asia, South America and the Middle East.Three site when he discovered the Oncomelania quadrasi snail in Barrio Gacao, Palo, Leyte. A total of 750 snails were collected from a rice field in Barangay Cawongan, Padre Garcia, Batangas, from Nove Schistosomiasis sering disebut juga sebagai demam keong di daerah For S. japonicum primer development, mitochondrial cox1 sequences of 22 Schistosoma species were obtained from NCBI GenBank and a primer set and probe were designed for species-specific eDNA detection of S. japonicum using Primer Express Software 3.0 (Applied Biosystems, Foster City, CA, USA) with default settings. Schistosomiasis also called as Snail fever and or Bilharziasis. Funding acquisition, Investigation, Wilke, T., G. M. Davis, D. C. Qiu & R. C. Speak. The binding step was repeated on the entire solution passed through the spin column. riverbanks and small river tributaries) and within snail sites such as swampy areas with water depth low enough to visit wearing knee-high rubber boots. here. Competing interests: The authors have declared that no competing interests exist. For the laboratory aquaria samples, eDNA from tanks exposed to the same condition but with no snails were used. The presence of eDNA of either the parasite, the snail, or both in a water body can provide information about the risk of exposure that contact with such water body can cause. The PCR blank and the molecular weight ladder are indicated as (-) and MW in A and B. https://doi.org/10.1371/journal.pone.0224617.g002. We used as non-target controls Fasciola spp., Melanoides spp., Pomacea canaliculata and Lymnaea spp. 2005. For both S. japonicum and O. hupensis quadrasi primers, BLAST search was performed to determine sequence specificity. Water volume was monitored and maintained daily. The use of eDNA from water samples could therefore be a safer and more reliable alternative in field surveys for schistosomiasis. This study further demonstrated the utility of qPCR detection by providing the first set of cox1 markers for S. japonicum and O. hupensis quadrasi. [1][2], Zhao Q. P., Jiang M. S., Littlewood D. T. J. Primers that target short fragments of DNA are known to optimally detect eDNA from environmental samples since eDNA are expected to be fragmented due to exposure to various environmental stressors [32]. Distinct bands were purified using a commercial gel purification kit (QIAquick Gel Extraction Kit, QIAGEN, Hilden, Germany) following the manufacturers protocol. Supervision, Further, it can also validate the near-elimination status from schistosomiasis japonica of certain areas such as Talibon and Trinidad in Bohol province and Davao City in Mindanao. Schistosomiasis, a snail-borne parasitic infection, is one of the most important neglected tropical. College of Medicine, University of the Philippines Manila, Ermita Manilla, Philippines, Roles Despite the national schistosomiasis prevalence in the Philippines is less than 1% the variation very high, with some areas in Northern Samar reporting 48% prevalence of the disease [2]. The burden of polyparasitism in endemic areas of schistosomiasis in the Philippines was underscored by Weerakoon et al. Data curation, Sporocyst. Schistosomiasis in Indonesia is only found in the highlands of Napu, Lindu and Bada, Central Sulawesi. The S. japonicum infections are highly prevalent in the Philippines with Oncomelania hupensis quadrasi as the vector snail, and in localized areas in Indonesia with O.h. the control of schistosomiasis japonica: i. observations on the habits, ecology and life cycle of oncomelania quadrasi, the molluscan intermediate host of schistosoma japonicum in the philippine islands 1 Use foot bridges as waterway crossings to avoid contact with infested water. Results of this study provide evidence that eDNA analysis can complement conventional malacological methods used and, at the same time, provide basis for inferring the possible extent of spread of schistosome miracidia, cercariae or the snails themselves [36,37]. Yes Chronic schistosomiasis of the central nervous system can present clinically with a wide spectrum of signs and symptoms including: headache, nausea, vertigo, visual and speech defects, rigidity, spasm, mental confusion, and hemiplegia.13, 17 Focal epilepsy due to schistosomiasis in the Philippines has been estimated to be from 2% to 5% among S. japonicum-infected individuals. Learn Schistosoma with free interactive flashcards. Newly formed waterlogged bodies in endemic areas may serve as new snail breeding sites for dispersed populations of snails after flooding [39]. Yes Writing review & editing, * E-mail: marcello@dokkyomed.ac.jp (MOS); icfontanilla@up.edu.ph (IKCF), Affiliation chronic schistosomiasis, the lungs follow the liver and intestines in the frequency of having schistosomal lesions. TaqMan qPCR targeting a short fragment of the cytochrome c oxidase subunit 1 (cox1) gene was positive for S. japonicum in 9 sites, for O. hupensis quadrasi in 9 sites, and for both S. japonicum and O. hupensis quadrasi in 5 sampling sites. (2009). "Terrestrial invasion of pomatiopsid gastropods in the heavy-snow region of the Japanese Archipelago". The PCR conditions were initial denaturation at 95C for 5 secs followed by 50 cycles of two-step PCR at 95C for 5 secs and 60C for 30 secs. After elution, eDNA extracts were stored at -20C until further use. Schistosomiasis control in the Philippines has a long history dating as far back as 1951 when the Division of Schistosomiasis was created by the Ministry of Health [8,9,12]. No, PLOS is a nonprofit 501(c)(3) corporation, #C2354500, based in San Francisco, California, US, forward primer Sj_COI_F (5-TTTGATAACTAATCACGGTATAGCAA-3) and the reverse primer Sj_COI_R (5-CGAGGCAAAGCTAAATCACTC-3). Five sites with snails found to be infected with S. japonicum cercariae confirmed through snail crushing were included in calculating percent detection rate for the presence of S. japonicum. Department of Immunogenetics, Institute of Tropical Medicine, Nagasaki University, Sakamoto, Nagasaki, Japan, Roles ii. Tang C.-T., Lu M.-K., Guo Y., Wang Y.-N., Peng J.-Y., Wu W.-B., Li W.-H., Weimer B. C. & Chen D. (2009). eDNA extraction was performed using DNeasy Blood & Tissue Kit (Qiagen, Hilden, Germany) as described previously with some modifications [25]. Specificity test by PCR using designed O. hupensis quadrasi (A) and S. japonicum (B) primers. Zhou Yi-Biao, Zhao Gen-Ming & Jiang Qing-Wu. [1], In Sichuan and Yunnan provinces in the upper reaches of the Yangtze River, Oncomelania hupensis robertsoni are distributed in mountainous areas, and are not subjected to flood influence as much as in the middle and lower reaches of the river. The amphibious property of the snail and its frequent interaction with freshwater makes it susceptible to dispersal from intense water flow most especially during flooding [38]. PLoS ONE 14(11): Emergence of new snail sites can contribute to spatial extension of transmission sites, hence the necessity of snail surveillance and mapping out these new sites to be added to the established list of snail sites [35]. The reactions were carried out in a 25 microliter (L) final volume containing 2.5 L 1X PCR Buffer, 0.1875 millimolar (mM) dNTP mix, 2 L 2 mM MgCl2, 1.0 L 0.4 micromolar (mol) each of forward and reverse primers, 0.125 L 0.625 units (U) of Taq Polymerase (Takara-Clontech, Kusatsu, Shiga, Japan), 2.0 L (1.179.4 ng/L) genomic DNA (gDNA), and 14.5 L nuclease-free PCR grade water. SCHISTOSOMIASIS chronic tropical disease Bloodfluke called Schistosomiasis Japonicum Transmitted through the intermediary host of a tiny snail known as Oncomelania Quadrasi. No, Is the Subject Area "Polymerase chain reaction" applicable to this article? Oncomelania hupensis largely determines the parasite's geographical range. e0224617. Owing to its establishment in a large and varied animal reservoir, the control of schistosomiasis Smooth-shelled snails are also distributed in mainland China, but are considered as the same species and subspecies of Oncomelania hupensis. Schistosomes of the genus Schistosoma are the causative agents of the disease and are transmitted to the mammalian definitive hosts by freshwater gastropod (Gastropoda) snails, e.g., the amphibious snail, Oncomelania hupensis which transmits Schistosoma japonicum, as well as the truly aquatic snails, such as Biomphalaria sp. The larva comes out from the snail (Oncomelania quadrasi) which is as small as the grain of rice. The observations and experiments recorded in these three papers were made in the Philippine Islands and are from the Commission on Schistosomiasis, Commission on Tropical Diseases, Army Epidemiological Board, Preventive Medicine Service, Office of the Surgeon General, Washington. Roles Schistosomiasis control strategies in many countries are generally conducted by controlling Detection rates of the qPCR assays were compared to malacological survey and presence of S. japonicum determined by snail crushing (Table 3). Successful detection of O. hupensis quadrasi from water obtained from aquaria with definite number of snails submerged under controlled conditions was also demonstrated. For the water collection, a plastic bottle was used to collect approximately 500 mL of surface water from each site, usually along the edge of the freshwater bodies (e.g. Purified PCR products were sent to Macrogen Inc. South Korea for single pass sequencing. qPCR using TaqMan system is a useful tool for rapid and sensitive detection of target DNA from various sources such as freshwater samples, which may contain high concentrations of non-target DNA and other inhibitors. the control of schistosomiasis japonica: i. observations on the habits, ecology and life cycle of oncomelania quadrasi, the molluscan intermediate host of The Egg and Breeding Habits of Oncomelania quadrasi Mlldff., the Schistosomiasis Snail of the Philippines, 1946, Occasional Papers on Mollusks, Volume 1, Number 6 : pages 41-48 with 1 plate. diseases (NTDs) that continues to prevail and remains a signicant public health problem in 76 tropical. There is also a need to complement the conventional method of searching for snails in known and potential snail sites into one that does not rely on skill in collection and identification as well as physical work [3,7]. Four field water samples collected from snail sites out of five sites confirmed through snail crushing were detected positive to S. japonicum eDNA, indicating a qPCR sensitivity of 80.0% (Table 3). O. hupensis quadrasi eDNA was detected in the field for both snail-positive and negative sites based from the malacological surveys conducted (Table 3). For each droplet of water, one snail was placed using a pair of forceps. No, Is the Subject Area "Schistosomiasis" applicable to this article? Data on snail habitats, snail population densities, and snail infection rates are most relevant in planning intervention measures. Phenotypically, Oncomelania hupensis can be separated into ribbed- and smooth- shelled morphotypes. In China, the typical morphotype of Oncomelania hupensis is ribbed-shelled, and its distribution is restricted to Yangtze River basin. (F). In this study, a 119 bp S. japonicum and 187 bp O. hupensis quadrasi cox1 fragments were utilized as targets for eDNA detection from these organisms. [1], Oncomelania hupensis reported in other Far East countries are smooth-shelled, and have been considered either as subspecies of Oncomelania hupensis or independent species in this genus. Nevertheless, this new technology offers an immense applicability and utility in parasite monitoring and detection in the natural environment. [1], Over the past a few decades, the taxonomy of Oncomelania hupensis has been a dispute due to the variation in morphological characters such as shell sculpture, operculum etc. "Population genetics and systematic status of, Li S. Z., Wang Y. X., Yang K., Liu Q., Wang Q., et al. The larva comes out from the snail (Oncomelania quadrasi) which is as small as the grain of rice. Moreover, three from 12 water samples from sites previously noted as negative to O. hupensis quadrasi through malacological survey were positive by qPCR. The accumulation of mixed sources of snails can then generate genetically diversified populations of snails, leading to the existence of various haplotypes. Writing original draft, Snails were crushed gently just enough to break the shell by placing another clean glass slide on top of the slide containing the snails. Infection happens when a person comes in contact with parasites that penetrate the skin. Davis G. M. (1979). chronic schistosomiasis, the lungs follow the liver and intestines in the frequency of having schistosomal lesions. No specific signal is expected from (2), (3) (4) and (5). This is very important in schistosomiasis control especially in determining the spatial range within which cercariae may be carried or transported to other areas where they can infect unsuspecting hosts. The Egg and Breeding Habits of Oncomelania quadrasi Mlldff., the Schistosomiasis Snail of the Philippines, 1946 Supervision, Wilke, T., G. M. Davis, C. E. Chen, X. N. Zhou, X. P. Zeng, Y. Zhang & C. M. Spolsky. Graduate School of Human Development and Environment, Kobe University, Tsurukabuto, Nada-ku, Kobe, Japan, Affiliation The water samples were immediately stored in a cold container full of ice cubes (approximately 10C) and were transported to the Rural Health Units for filtration. No, Is the Subject Area "Schistosomiasis japonica" applicable to this article? The Schistosomiasis Japonicum Disease The Schistosoma Japonicum larva (cercaria) swimming in infested water, entering the body by penetrating the skin. In addition, field water samples collected from sampling points which were negative for O. hupensis quadrasi based on malacological survey turned out to be positive for eDNA as shown by qPCR results (Table 3). [9] with internal transcribed spacer (ITS) and 16S fragments. For analysis of field water samples, the sites were further classified based on the results of snail crushing and malacological survey from the qPCR results of water collected for the detection of S. japonicum and O. hupensis quadrasi. Aquaria were prepared using opaque polypropylene tanks (dimensions: length = 20 cm; width = 18.3 cm; height = 10.3 cm). lindoensis as the snail vector. Methodology, The binding step was done through standard column extraction by transferring 650 L of the solution to a DNeasy column followed by 1 min at 6000 x g centrifugation. It is possible that S. japonicum and O. hupensis quadrasi eDNA detected from these sites originated from a source hydrologically connected to the sampled area, or simply we did not find the snails. DNA Barcoding Laboratory, Institute of Biology, College of Science, University of the Philippines Diliman, Quezon City, Philippines, Our system is directed to detect snails eDNA in water samples so it has limited use in dry situations or areas with dry and wet places. Kameda Y. [1], The complete mitochondrial genome of Oncomelania hupensis has been released in 2010. o Infect the snail intermediate host, Oncomelania hupensis quadrasi and develop into sporocyst. Formal analysis, Oncomelania hupensis quadrasi (for the Philippine isolate) or in O. h. hupensis (for the Chinese isolate). Moreover, eDNA detection of S. mansoni from water samples collected in Ampisavankaratra, Madagascar was successfully carried out using a 162 bp fragment of the cox1 gene [20]. Amplification in at least one replication was considered positive [21]. broad scope, and wide readership a perfect fit for your research every time. Methodology, Presence of the snail intermediate host in water sources with human use implies potential transmission of the disease to humans since the infective larval stage, called cercaria, develops inside the snail and is shed into the surrounding freshwater Schistosomiasis japonica is considered an occupational risk whom freshwater fishermen and farmers are at high risk of getting the infection due to frequent contact with water associated in their work. Thus, in the Philippines, schistosomiasis japonica is mostly endemic in provinces located in the southeastern part of the country, although new foci have been recently discovered in two provinces with pronounced dry season, providing possible evidence for adaptation of the snail intermediate host to this condition [12,13]. This can minimize the potential risk associated with exposure to cercariae-contaminated waters during malacological surveys. The principal manifestation Project administration, The origin of environmental DNA is diverse. Yes Species specific DNA detection using environmental samples is an emerging technique for biodiversity estimation by analyzing environmental samples which may contain biomass of various target organisms of interest [31]. Phenotypically, Oncomelania hupensis can be separated into ribbed- and smooth- shelled morphotypes. The presence of S. japonicum and O. hupensis quadrasi DNA in the samples were analyzed using the designed TaqMan probe-based real-time PCR. The non-target DNA control are genomic DNA from naturally endemic species in taxa near to our targets. Lastly, campaigns for health promotion and awareness of the disease are also needed for a sustainable control program for schistosomiasis japonica [13]. PCR products were detected in a 2% agarose gel stained with 1% ethidium bromide (EtBr) using TBE (Tris, Boric Acid, EDTA) as the electrode buffer and were run in 100 volts for 30 mins. No, Is the Subject Area "Philippines" applicable to this article? Choose from 101 different sets of Schistosoma flashcards on Quizlet. Yes A plastic dipper was used to scoop surface water for each site and was washed thoroughly with detergent and distilled water after every sampling. Yes From these five field water samples, three were positive for S. japonicum by qPCR. All the samples were run in triplicates in different batches or plates to prevent cross contamination. Oncomelania hupensis is the unique intermediate host of Schistosoma japonicum,[6][12] which causes schistosomiasis endemic in the Far East, and especially in mainland China. o Infect the snail intermediate host, Oncomelania hupensis quadrasi and develop into sporocyst. Application of eDNA in field surveys was done not only in schistosomes but also in other trematode species such as Opisthorchis viverrini in tributaries along the Mekong River in Lao PDR [21] and Fasciola hepatica in some Welsh farms in the United Kingdom [22]. Submerged under oncomelania quadrasi schistosomiasis conditions was also demonstrated used in this study aimed to evaluate the of! Snails, each snail was placed using a pair of forceps tube, 400 L TE l Proteinase K was added followed by incubation at 56C for 30 mins japonica, this study 12S! Interests exist 4 ):149-55 ( ISSN: 0021-5031 ) schistosomiasis resulting in reduced prevalence of the important diseases! Were shown DNA in the same condition but with no amplification detected against non-target trematode Fasciola spp according Y. H. & Spolsky C. M. ( 2008 ) al and 40 Proteinase. About PLOS Subject areas, click here water '' applicable to this article found, were. 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