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Martini M, Cioli T, Romig T, Gagliardo A, Giunchi D, Zaccaroni M, Massolo A. Echinococcus multilocularis infection affects risk-taking behaviour in Microtus arvalis: adaptive manipulation? Parasitology 2024; 151:650-656. [PMID: 38766838 PMCID: PMC11474016 DOI: 10.1017/s0031182024000507] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/04/2023] [Revised: 04/22/2024] [Accepted: 04/24/2024] [Indexed: 05/22/2024]
Abstract
Manipulation of host behaviour by parasites to enhance transmission to the next host is a fascinating phenomenon that has interested scientists since the 1970s. It has been proposed that infection with the cestode Echinococcus multilocularis produces an impairment of the antipredatory behaviour in the rodent intermediate host common vole, Microtus arvalis, which may facilitate transmission of the tapeworm to the canid final host. In this study, we observed the behaviour of infected common voles at 12 weeks post-infection, when protoscoleces production and maturation commonly occurs, in order to assess behavioural changes compared to uninfected controls, that might ease predation in the wild. Infected and uninfected voles were monitored for 24 h to observe their spontaneous activity. In addition, the next day, both infected and uninfected voles were subjected to 4 different behavioural tests: open field test, barrier test, platform test and air-puff test in a running wheel. No significant difference between uninfected and infected voles emerged during the behavioural tests. However, observation of spontaneous activity revealed that infected voles increased their feeding frequency and spent significantly more time above bedding even when not eating, compared to the uninfected controls. In the wild, these behavioural changes increase the animals exposure to predators, raising their chance of becoming prey. These findings are the first direct evidence consistent with behavioural manipulation by E. multilocularis on common voles.
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Affiliation(s)
- Matilde Martini
- Department of Biology, Ethology Unit, University of Pisa, Pisa, Italy
- Department of Biological, BIOME Unit, Geological and Environmental Sciences, University of Bologna, Bologna, Italy
| | - Teila Cioli
- Department of Biology, Ethology Unit, University of Pisa, Pisa, Italy
| | - Thomas Romig
- Parasitology Unit, Institute of Biology, University of Hohenheim, Stuttgart, Germany
| | - Anna Gagliardo
- Department of Biology, Ethology Unit, University of Pisa, Pisa, Italy
| | - Dimitri Giunchi
- Department of Biology, Ethology Unit, University of Pisa, Pisa, Italy
| | - Marco Zaccaroni
- Department of Biology, University of Florence, Florence, Italy
| | - Alessandro Massolo
- Department of Biology, Ethology Unit, University of Pisa, Pisa, Italy
- Faculty of Veterinary Medicine, University of Calgary, Calgary, AB, Canada
- UMR CNRS 6249 Chrono-environnement, Université Franche-Comté, Besançon, France
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Corrêa LQ, do Couto BP, de Carvalho EFG, de Sousa JEN, da Silva Ribeiro V, Gonzaga HT, Costa-Cruz JM. Route of dexamethasone administration influences parasite burden in Strongyloides hyperinfection model. J Parasit Dis 2023; 47:520-526. [PMID: 37520210 PMCID: PMC10382442 DOI: 10.1007/s12639-023-01595-6] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/15/2023] [Accepted: 05/06/2023] [Indexed: 08/01/2023] Open
Abstract
Rodents infected with Strongyloides venezuelensis are experimental models applied to strongyloidiasis research. This study evaluated oral and subcutaneous dexamethasone (DEX) treatments to establish immunosuppression in an experimental model of Strongyloides hyperinfection. Rattus norvegicus Wistar were divided: G I (-): untreated and uninfected animals, G II (+): untreated and infected, G III (o -) orally treated and uninfected, G IV (o +) orally treated and infected, G V (sc -) subcutaneously treated and uninfected, G VI (sc +) subcutaneously treated and infected. For oral administration, DEX was diluted in sterile water (5 µg/ml) and made available to the animals on intervals in experimental days - 5-0, 8-13 and 21-26. For subcutaneous administration, animals received daily injections of DEX disodium phosphate (2 mg/kg). Infection was established by the subcutaneous inoculation of 3000 S. venezuelensis filarioid larvae. Groups were evaluated by egg per gram of feces and parasite females counts and IgG, IgG1 and IgG2a detection. GIV (o +) had egg peaks count on days 13 and 26 and maintained egg elimination until the last experimental day. Parasitic females recovery at day 30 was significantly higher in G IV (o +) when compared to G VI (sc +). Levels of IgG, IgG1 and IgG2a of all groups, except the positive control GII (+), were below the detection threshold. Pharmacological immunosuppression induced by oral administration of DEX produced high parasitic burden, and is a noninvasive method, useful to establish immunosuppression in strongyloidiasis hyperinfection model in rats.
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Affiliation(s)
- Luisa Queiroz Corrêa
- Laboratório de Diagnóstico de Parasitoses, Instituto de Ciências Biomédicas, Universidade Federal de Uberlândia, Uberlândia, Minas Gerais Brazil
| | - Bruna Patrícia do Couto
- Laboratório de Diagnóstico de Parasitoses, Instituto de Ciências Biomédicas, Universidade Federal de Uberlândia, Uberlândia, Minas Gerais Brazil
| | - Edson Fernando Goulart de Carvalho
- Laboratório de Diagnóstico de Parasitoses, Instituto de Ciências Biomédicas, Universidade Federal de Uberlândia, Uberlândia, Minas Gerais Brazil
| | - José Eduardo Neto de Sousa
- Laboratório de Diagnóstico de Parasitoses, Instituto de Ciências Biomédicas, Universidade Federal de Uberlândia, Uberlândia, Minas Gerais Brazil
| | - Vanessa da Silva Ribeiro
- Laboratório de Diagnóstico de Parasitoses, Instituto de Ciências Biomédicas, Universidade Federal de Uberlândia, Uberlândia, Minas Gerais Brazil
| | - Henrique Tomaz Gonzaga
- Laboratório de Diagnóstico de Parasitoses, Instituto de Ciências Biomédicas, Universidade Federal de Uberlândia, Uberlândia, Minas Gerais Brazil
| | - Julia Maria Costa-Cruz
- Laboratório de Diagnóstico de Parasitoses, Instituto de Ciências Biomédicas, Universidade Federal de Uberlândia, Uberlândia, Minas Gerais Brazil
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Proof-of-Concept Preclinical Use of Drosophila melanogaster in the Initial Screening of Immunomodulators. Sci Pharm 2022. [DOI: 10.3390/scipharm90010011] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 02/01/2023] Open
Abstract
Drug discovery is a complex process, and the use of a comprehensive approach is deemed necessary to discover new chemical entities with novel mechanisms of action. This research was carried out to determine whether Drosophila melanogaster can serve as an appropriate model organism in the initial screening of drug candidates with immunomodulatory activities. To test this, we performed phenotypic assay and molecular analysis to investigate the immunomodulatory activities of aspirin, dexamethasone, curcumin, and epigallocatechin gallate (EGCG), that have been reported to yield such effects in the mammalian model system. In vivo survival analysis demonstrated that all drugs/compounds were relatively safe at the tested concentrations. In the infection assay, curcumin and EGCG showed a protective signature to bacterial infection in flies lacking Toll-mediated immune responses. Furthermore, dexamethasone and aspirin, drugs with immunosuppressive activity, could improve the survival of PGRP-LBΔ mutant flies with hyperactivated immune system. These phenotypes were supported by RT-qPCR-based molecular analysis, revealing that drugs/compounds used in this study could modulate the expression level of genes related to the immune system. In conclusion, while curcumin and EGCG could promote the improvement of fly survival against infection, aspirin and dexamethasone were able to suppress overactivation of immune responses in D. melanogaster. These results are in line with the ones observed in the mammalian model system, further emphasizing the notion that flies would serve as a prospective model organism in the initial screening of drug candidates for their immunomodulatory activities prior to further checking in the mammalian animal models. In the end, this will reduce the use of mammalian animal models for preliminary experiments in an effort to discover/repurpose drugs with immunomodulatory activity.
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Gottstein B, Deplazes P. Alveolar echinococcosis: what triggers emergence in North America, Central Europe and Asia? Curr Opin Infect Dis 2021; 34:440-446. [PMID: 34524197 DOI: 10.1097/qco.0000000000000765] [Citation(s) in RCA: 7] [Impact Index Per Article: 1.8] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/16/2022]
Abstract
PURPOSE OF REVIEW Infection with the larval (metacestode) stage of Echinococcus multilocularis causes alveolar echinococcosis (AE), a serious hepatic disorder. The parasite has increased its infection extensity in wildlife and domestic dogs, mainly due to urbanization and spatial extension of wildlife hosts in Europe, Asia as well as North America, resulting in emerging infection risk for humans. RECENT FINDINGS In hyperendemic areas such as Kyrgyzstan and China, ecological and socioeconomic changes have been associated with the unpredictable increase of AE cases. In North America, the appearance of the European-like genotype is of concern. In Europe, the annual increase of human case numbers reached a plateau even in hyperendemic situations. Therefore, we conclude that most of the exposed individuals are resistant to parasite invasion and/or to disease development. Thus, AE develops in a few healthy individuals, but preferentially in immunosuppressed patients. SUMMARY In the future, improved diagnostic strategies will allow more precise estimations of transmission routes including the role of food, water and direct dog contact, which should yield improved public health recommendations. Finally, understanding protective innate and acquired immune mechanisms as well as parasite-driven immune-evasion processes will be essential to develop curative therapies in nonoperable patients and, futuristically, appropriate vaccines.
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Affiliation(s)
- Bruno Gottstein
- Institute of Infectious Diseases, Faculty of Medicine, University of Bern, Bern
| | - Peter Deplazes
- Institute of Parasitology, Vetsuisse Faculty and Faculty of Medicine, University of Zurich, Zurich, Switzerland
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Early-phase migration dynamics of Echinococcus multilocularis in two mouse strains showing different infection susceptibilities. Int J Parasitol 2021; 51:893-898. [PMID: 33991567 DOI: 10.1016/j.ijpara.2021.04.001] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/03/2021] [Revised: 04/08/2021] [Accepted: 04/09/2021] [Indexed: 11/24/2022]
Abstract
The early-phase migration dynamics of Echinococcus multilocularis in the intermediate hosts remain largely unknown. We compared the parasite burden in the intestine, liver and faeces of DBA/2 and C57BL/6 mouse strains using parasite-specific quantitative PCR. Our results indicated that the parasites invaded mainly from the middle segments of the small intestine and completed migration to the liver within 24 h p.i. C57BL/6 mice had lower parasite DNA burdens in the intestine and liver but higher in the faeces than DBA/2 mice, suggesting that parasite invasion of the intestine may be a critical stage regulating susceptibility to E. multilocularis infection in mice.
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Joekel DE, Nur S, Monné Rodriguez J, Kronenberg PA, Kipar A, LeibundGut-Landmann S, Deplazes P. Agranulocytosis leads to intestinal Echinococcus multilocularis oncosphere invasion and hepatic metacestode development in naturally resistant Wistar rats. Parasitology 2021; 148:53-62. [PMID: 33087186 PMCID: PMC11010136 DOI: 10.1017/s0031182020002012] [Citation(s) in RCA: 4] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/03/2020] [Revised: 08/28/2020] [Accepted: 08/30/2020] [Indexed: 12/12/2022]
Abstract
Susceptibility to Echinococcus multilocularis infection considerably varies among intermediate (mostly rodents) and dead-end host species (e.g. humans and pig), in particular regarding intestinal oncosphere invasion and subsequent hepatic metacestode development. Wistar rats are highly resistant to infection and subsequent diseases upon oral inoculation with E. multilocularis eggs, however, after immunosuppressive treatment with dexamethasone, rats become susceptible. To address the role of the cellular innate immunity, Wistar rats were individually or combined depleted of natural killer (NK) cells, macrophages (MΦ) and granulocytes (polymorphonuclear cells, PMN) prior to E. multilocularis egg inoculation. Although NK cell and MΦ depletion did not alter the resistance status of rats, the majority of PMN-depleted animals developed liver metacestodes within 10 weeks, indicating that PMN are key players in preventing oncosphere migration and/or development in Wistar rats. In vitro studies indicated that resistance is not caused by neutrophil reactive oxygen species or NETosis. Also, light microscopical examinations of the small intestine showed that oral inoculation of E. multilocularis eggs does not elicit a mucosal neutrophil response, suggesting that the interaction of oncospheres and neutrophils may occur after the former have entered the peripheral blood. We suggest to consider granulocytes as mediators of resistance in more resistant species, such as humans.
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Affiliation(s)
- Deborah E. Joekel
- Institute of Parasitology, Vetsuisse Faculty, University of Zurich, Winterthurerstrasse 266a, 8057 Zurich, Switzerland
| | - Selim Nur
- Section of Immunology, Institute of Virology, Vetsuisse Faculty, University of Zurich, Winterthurerstrasse 266a, 8057Zurich, Switzerland
| | - Josep Monné Rodriguez
- Laboratory for Animal Model Pathology (LAMP), Institute of Veterinary Pathology, Vetsuisse Faculty, University of Zurich, Winterthurerstrasse 268, 8057Zurich, Switzerland
| | - Philipp A. Kronenberg
- Institute of Parasitology, Vetsuisse Faculty, University of Zurich, Winterthurerstrasse 266a, 8057 Zurich, Switzerland
- Graduate School for Cellular and Biomedical Sciences, University of Bern, Mittelstrasse 43, 3012Bern, Switzerland
| | - Anja Kipar
- Laboratory for Animal Model Pathology (LAMP), Institute of Veterinary Pathology, Vetsuisse Faculty, University of Zurich, Winterthurerstrasse 268, 8057Zurich, Switzerland
| | - Salomé LeibundGut-Landmann
- Section of Immunology, Institute of Virology, Vetsuisse Faculty, University of Zurich, Winterthurerstrasse 266a, 8057Zurich, Switzerland
| | - Peter Deplazes
- Institute of Parasitology, Vetsuisse Faculty, University of Zurich, Winterthurerstrasse 266a, 8057 Zurich, Switzerland
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Pathology of Echinococcosis: A Morphologic and Immunohistochemical Study on 138 Specimens With Focus on the Differential Diagnosis Between Cystic and Alveolar Echinococcosis. Am J Surg Pathol 2020; 44:43-54. [PMID: 31567204 DOI: 10.1097/pas.0000000000001374] [Citation(s) in RCA: 28] [Impact Index Per Article: 5.6] [Reference Citation Analysis] [Abstract] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/11/2022]
Abstract
Infection of humans by the larval stage of the tapeworms Echinococcus granulosus sensu lato or Echinococcus multilocularis causes the life-threatening zoonoses cystic echinococcosis (CE) and alveolar echinococcosis (AE). Although cystic liver lesions are a hallmark of both diseases, course, prognosis, and patients' management decisively differ between the two. The wide and overlapping spectrum of morphologies and the limited availability of ancillary tools are challenges for pathologists to reliably diagnose and subtype echinococcosis. Here, we systematically and quantitatively recorded the pathologic spectrum in a clinically and molecularly defined echinococcosis cohort (138 specimens from 112 patients). Immunohistochemistry using a novel monoclonal antibody (mAbEmG3) was implemented, including its combined application with the mAbEm2G11. Six morphologic criteria sufficiently discriminated between CE and AE: size of smallest (CE/AE: >2/≤2 mm) and largest cyst (CE/AE: >25/≤25 mm), thickness of laminated layer (CE/AE: >0.15/≤0.15 mm) and pericystic fibrosis (CE/AE: >0.6/≤0.6 mm), striation of laminated layer (CE/AE: moderate-strong/weak), and number of cysts (CE/AE: ≤9/>9). Combined immunohistochemistry with mAbEm2G11 (E. multilocularis specific) and mAbEmG3 (reactive in AE and CE) was equally specific as and occasionally more sensitive than polymerase chain reaction. On the basis of these findings, we developed a diagnostic algorithm for the differential diagnosis of echinococcosis. In summary, we have not only identified the means to diagnose echinococcosis with greater certainty, but also defined morphologic criteria, which robustly discriminate between CE and AE. We expect our findings to improve echinococcosis diagnostics, especially of challenging cases, beneficially impacting the management of echinococcosis patients.
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Beerli O, Guerra D, Baltrunaite L, Deplazes P, Hegglin D. Microtus arvalis and Arvicola scherman: Key Players in the Echinococcus multilocularis Life Cycle. Front Vet Sci 2017; 4:216. [PMID: 29326950 PMCID: PMC5733337 DOI: 10.3389/fvets.2017.00216] [Citation(s) in RCA: 16] [Impact Index Per Article: 2.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/05/2017] [Accepted: 11/29/2017] [Indexed: 11/30/2022] Open
Abstract
A broad range of rodent species are described as potential intermediate hosts for Echinococcus multilocularis, a wide-spread zoonotic cestode causing alveolar echinococcosis. However, little is known about the relative contribution of these species for parasite reproduction and the maintenance of its life cycle. In a comparative study in a high endemic region in Zurich, Switzerland, we investigated prevalence rates and fertility of E. multilocularis in the most abundant vole species as well as the predation rate of foxes on these species. To ensure that the fox families had access to different vole species and that these voles were exposed to the same environmental contamination with parasite eggs, we selected eight study plots where at least two rodent species co-occurred. The parasite prevalence in Microtus arvalis [11.0%, confidence intervals (CI) 8.9–13.4] was significantly higher than in Arvicola scherman (5.3%, 3.9–7.1) and Myodes glareolus (3.9%, 2.0–6.7). None of the, only 29 individuals of, Microtus agrestis was infected (0%, 0.0–9.8) and the species was excluded for further analyses. Logistic regression models for the prevalences revealed significant differences between nearby study plots and higher infection rates for females, heavier individuals, and individuals trapped during spring, when the prevalence in M. arvalis peaked up to 65% (CI 50–79) in one plot. Furthermore, we detected significantly higher percentages of fertile infections in M. arvalis and M. glareolus than in A. scherman (OR 11.2 and 6.4, respectively) and a significantly higher protoscolex number in M. glareolus (median 100,000) than in M. arvalis (13,500) and A. scherman (4,290). The most abundant fox prey remains were of the genera Microtus (12.3%, CI 8.4–17.2) and Arvicola (11.5%, 7.7–16.3), whereas Myodes was never recorded as prey (0.0–1.3%). We conclude that M. arvalis and to a lesser extent A. scherman can be regarded as key intermediate hosts in Western and Central European high-endemic regions whereas M. glareolus and M. agrestis play a marginal role. We, therefore, postulate that distribution models of these species could contribute to predict parasite occurrence on a more detailed spatial scale than models of the distribution of foxes which have a very broad and uniform distribution.
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Affiliation(s)
- Olivia Beerli
- Institute of Parasitology, University of Zurich, Zurich, Switzerland
| | - Diogo Guerra
- Institute of Parasitology, University of Zurich, Zurich, Switzerland
| | - Laima Baltrunaite
- Laboratory of Mammalian Ecology, Nature Research Centre, Vilnius, Lithuania
| | - Peter Deplazes
- Institute of Parasitology, University of Zurich, Zurich, Switzerland
| | - Daniel Hegglin
- Institute of Parasitology, University of Zurich, Zurich, Switzerland
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Zhang RQ, Chen XH, Wen H. Improved experimental model of hepatic cystic hydatid disease resembling natural infection route with stable growing dynamics and immune reaction. World J Gastroenterol 2017; 23:7989-7999. [PMID: 29259374 PMCID: PMC5725293 DOI: 10.3748/wjg.v23.i45.7989] [Citation(s) in RCA: 10] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 07/21/2017] [Revised: 10/06/2017] [Accepted: 10/17/2017] [Indexed: 02/06/2023] Open
Abstract
AIM To investigate a safer way to set up the disease model of cystic echinococcosis without contamination risk and develop a novel experimental murine model of hepatic cystic echinococcosis.
METHODS C57B/6 mice were injected with human protoscolices of three different concentrations via the portal vein. The mice were followed for 10 mo by ultrasound, gross anatomy, and pathological and immunological examinations. The protoscolex migration in the portal vein, hydatid cyst growth, host immune reaction, and hepatic histopathology were examined periodically.
RESULTS The infection rates in the mice in the high, medium, and low concentration groups were 90%, 100%, and 63.6%, respectively. The protoscolices migrated in the portal vein with blood flow, settled in the liver, and developed into orthotopic hepatic hydatid cysts, resembling the natural infection route and course.
CONCLUSION We have established an improved experimental model of hepatic cystic echinococcosis with low biohazard risk but stable growing dynamics and immune reaction. It is especially useful for new anti-parasite medication trials against hydatid disease.
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Affiliation(s)
- Rui-Qing Zhang
- Hepatobiliary and Hydatid Department, Digestive and Vascular Surgery Centre, Xinjiang Key Laboratory of Echinococcosis, The First Affiliated Hospital of Xinjiang Medical University, Urumqi 830011, Xinjiang Uygur Autonomous Region, China
| | - Xin-Hua Chen
- Department of Hepatobiliary and Pancreatic Surgery, The First Affiliated Hospital, Zhejiang University, Hangzhou 310000, Zhejiang Province, China
- Key Laboratory of Combined Multi-organ Transplantation, Ministry of Public Health, The First Affiliated Hospital, Zhejiang University, Hangzhou 310000, Zhejiang Province, China
- Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, The First Affiliated Hospital, Zhejiang University, Hangzhou 310000, Zhejiang Province, China
| | - Hao Wen
- Hepatobiliary and Hydatid Department, Digestive and Vascular Surgery Centre, Xinjiang Key Laboratory of Echinococcosis, The First Affiliated Hospital of Xinjiang Medical University, Urumqi 830011, Xinjiang Uygur Autonomous Region, China
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