RESUMEN
OBJECTIVES: This study aimed to assess the diagnostic potential of cell-free DNA (cfDNA) and cell-free miRNA (cf-miRNA) for distinguishing between Healthy, asymptomatic opisthorchiasis viverrini and cholangiocarcinoma in a preliminary manner. METHODS: In this study, 36 participants were enrolled into three health status groups: a healthy control group (HC), Opisthorchis viverrini-infected group (OV), and a cholangiocarcinoma group (CCA), each comprising 12 participants. Concentration measurements of cfDNA and cf-miRNA from plasma were conducted. Additionally, ultra-low-pass whole-genome sequencing (ULP-WGS) was employed to investigate DNA alterations. RESULTS: The study revealed a significant elevation in plasma cfDNA concentration in the cholangiocarcinoma (CCA) group compared to healthy controls (HC) and Opisthorchis viverrini-infected (OV) groups (P < 0.001). The cfDNA concentration demonstrated a sensitivity of 75.00% and specificity of 95.83% for differentiating cholangiocarcinoma, with a cut-off of > 30.50 ng/ml plasma. Likewise, the concentration of cf-miRNA in the CCA group significantly differed from that in the HC and OV groups, demonstrating a sensitivity of 83.33% and specificity of 95.83% with a cut-off set at > 70.50 ng/ml plasma. Furthermore, a positive correlation between plasma concentrations of cfDNA and cf-miRNA suggests a potential relationship between these two biomarkers. These findings indicated the diagnostic potential of cfDNA and cf-miRNA in distinguishing cholangiocarcinoma, emphasizing their role as promising biomarkers for further investigation and clinical applications. CONCLUSION: Elevated plasma concentrations of cfDNA and cf-miRNA could serve as potential diagnostic tools for distinguishing cholangiocarcinoma from other conditions. cf-miRNA was superior to cfDNA in terms of sensitivity.
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Neoplasias de los Conductos Biliares , Ácidos Nucleicos Libres de Células , Colangiocarcinoma , MicroARNs , Opistorquiasis , Opisthorchis , Animales , Humanos , Opistorquiasis/complicaciones , Opistorquiasis/diagnóstico , MicroARNs/genética , Colangiocarcinoma/diagnóstico , Colangiocarcinoma/genética , Biomarcadores , Conductos Biliares Intrahepáticos/patología , Neoplasias de los Conductos Biliares/diagnóstico , Neoplasias de los Conductos Biliares/genéticaRESUMEN
This study aimed to identify the recent risk factors for Opisthorchis viverrini infection and cholangiocarcinoma (CCA) to improve disease prevention. The participants were divided into the following 3 groups based on their health status: healthy control (nonOV and nonCCA), those with O. viverrini infection (OV), and those with CCA. A questionnaire was used to explore their lifestyle and behaviors. Multivariate logistic regression and backward elimination were used to identify the significant risk factors. The results showed that the significant risk factors for both O. viverrini infection and CCA were age>50 years (odd ratio (OR)=8.44, P<0.001, 95% confidence intervals (CI) 2.98-23.90 and OR=43.47, P=0.001, 95% CI 14.71-128.45, respectively) and raw fish consumption (OR=8.48, P< 0.001, 95% CI 3.18-22.63 and OR=3.15, P=0.048, 95% CI 1.01-9.86, respectively). A history of O. viverrini infection was identified as an additional risk factor for CCA (OR=20.93, P=0.011, 95% CI 2.04-215.10). This study provided an update on the risk factors for O. viverrini infection and CCA. Asymptomatic patients with O. viverrini infection, particularly those>50 years old, should be carefully monitored to prevent CCA.
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Neoplasias de los Conductos Biliares , Colangiocarcinoma , Opistorquiasis , Opisthorchis , Animales , Humanos , Persona de Mediana Edad , Opistorquiasis/complicaciones , Tailandia/epidemiología , Neoplasias de los Conductos Biliares/epidemiología , Colangiocarcinoma/epidemiología , Factores de Riesgo , Conductos Biliares Intrahepáticos/patologíaRESUMEN
In Southeast Asian countries, nitrosamine compounds and the liver fluke Opisthorchis viverrini have long been identified as carcinogens for cholangiocarcinoma (CHCA). In order to effectively treat O. viverrini infections and prevent the development of CHCA, methods for disease detection are needed. This study aims to identify biomarkers for O. viverrini infection and CHCA. In the discovery phase, technical triplicates of five pooled plasma pools (10 plasma each) of healthy control subjects (noOVCCA), O. viverrini subjects (OV), and cholangiocarcinoma subjects (CCA), underwent solution-based digestion, with the label-free method, using a Thermo Scientific™ Q Exactive™ HF hybrid quadrupole-Orbitrap mass spectrometer and UltiMate 300 LC systems. The noOVCCA, OV, and CCA groups demonstrated different profiles and were clustered, as illustrated by PCA and heat map analysis. The STRING and reactome analysis showed that both OV and CCA groups up-regulated proteins targeting immune system-related proteins. Differential proteomic profiles, S100A9, and polymeric immunoglobulin receptor (PIGR) were specifically expressed in the CCA group. During the validation phase, another 50 plasma samples were validated via the PIGR sandwich ELISA. Using PIGR >1.559 ng/ml as a cut-off point, 78.00% sensitivity, 71.00% specificity, and AUC = 0.8216, were obtained. It is sufficient to differentially diagnose cholangiocarcinoma patients from healthy patients and those with Opisthorchiasis viverrini. Hence, in this study, PIGR was identified and validated as a potential biomarker for CHCA. Plasma PIGR is suggested for screening CHCA, especially in an endemic region of O. viverrini infection.
RESUMEN
Among the snail species acting as hosts for medically significant trematodes, only three taxa of Bithynia are responsible for transmitting the carcinogenic liver fluke Opisthorchis viverrini to humans in different geographical areas. Although B. siamensis goniomphalos is the primary species responsible for O. viverrini transmission in endemic areas, B. siamensis siamensis and B. funiculata remain potential hosts for transmission. This study objects to determine the susceptibility of B. siamensis siamensis and B. funiculata to O. viverrini to assess the risk of O. viverrini transmission in non-endemic areas. The snails of both species were first introduced to O. viverrini eggs, after which O. viverrini infection was investigated using specific PCR primers after a period of 1, 7, 14, 28, and 56 days post-infection (dpi). Opisthorchis viverrini infection in both B. siamensis siamensis and B. funiculata was high in the early period (1 and 7 dpi) while decreasing over time. It was also shown that the odds of susceptibility to O. viverrini infection in B. siamensis siamensis were 64.5% higher relative to the odds of susceptibility in B. funiculata (P < 0.05). Results of this study provide an early insight into the Bithynia-Opisthorchis relationship and thus have great potential to assess risk and raise awareness of opisthorchiasis in non-endemic regions, especially in regions endemic for B. siamensis siamensis.
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Opistorquiasis , Opisthorchis , Humanos , Animales , Opistorquiasis/epidemiología , Opistorquiasis/veterinaria , Caracoles , Tailandia/epidemiologíaRESUMEN
Several trematodes including Opisthorchis viverrini utilize Bithynia siamensis goniomphalos as a snail intermediate host in their life cycles. In order to capture a comprehensive range of hostparasite interactions and their transmission dynamic patterns, B. s. goniomphalos were sampled monthly over 4 consecutive years in an irrigated paddy-field habitat in northeast Thailand. Using a standard cercarial shedding method, a high diversity of trematodes (17 types) was recovered. Virgulate xiphidiocercariae were the most prevalent (7.84%) followed by O. viverrini (0.71%). In addition to seasonal and environmental factors, the quantity of irrigation water for rice cultivation correlated with transmission dynamics of trematodes in B. s. goniomphalos. The peak prevalence of all trematode infections combined in the snails shifted from the cool-dry season in 20102012 to the hot-dry season in 2013 associated with an increasing quantity of water irrigation. A low frequency of mixed trematode infections was found, indicating that the emergence of virgulate cercariae, but not of O. viverrini, was negatively impacted by the presence of other trematodes in the same snail. Taken together, the observed results suggest that interactions between host and parasite, and hence transmission dynamics, depend on specific characteristics of the parasite and environmental factors including irrigated water for rice cultivation.
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Opisthorchis , Infecciones por Trematodos , Animales , Cercarias , Dinámica Poblacional , Caracoles/parasitología , Tailandia/epidemiología , Infecciones por Trematodos/epidemiologíaRESUMEN
The human liver fluke Opisthorchis viverrini (Ov), the primary risk factor for cholangiocarcinoma (CHCA), is a parasite endemic to southeast Asian countries. With no effective treatments for CHCA currently available, early diagnosis and treatment of Ov infection remains the only practical method for the prevention of CHCA. In this study, plasma phosphoproteomes of patients in the non-Ov infection, non-cholangiocarcinoma subject group (non-OVCCA), the asymptomatic Ov infected group (OV), and the CHCA group (CCA), were investigated to identify potential biomarkers for Ov infection and CHCA. The AKT signalling pathway was found to be up-regulated. Phosphatidylinositol 4,5-bisphosphate 3-kinase catalytic subunit beta isoform (PIK3CB), an upstream signalling molecule, was selected as a potential biomarker and evaluated using indirect enzyme-linked immunosorbent assay (ELISA). Results demonstrated evidence that levels of PIK3CB in both the OV group and CCA group was statistically different compared to the non-OVCCA group (P < 0.01). However, the levels of PIK3CB between the OV group and the CCA group were found not to be statistically different. Sensitivity and specificity for OV using OD450 cut-off at >1.570 was 76 and 72%, respectively. For CCA, sensitivity and specificity using OD450 cut-off at >1.398 was 68 and 76%, respectively. Application of indirect ELISA detecting plasma PIK3CB will be of great benefit for screening of opisthorchiasis and CHCA.
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Neoplasias de los Conductos Biliares , Colangiocarcinoma , Opistorquiasis , Opisthorchis , Animales , Neoplasias de los Conductos Biliares/diagnóstico , Neoplasias de los Conductos Biliares/epidemiología , Neoplasias de los Conductos Biliares/metabolismo , Conductos Biliares Intrahepáticos/metabolismo , Conductos Biliares Intrahepáticos/parasitología , Conductos Biliares Intrahepáticos/patología , Biomarcadores/análisis , Dominio Catalítico , Colangiocarcinoma/diagnóstico , Colangiocarcinoma/etiología , Colangiocarcinoma/metabolismo , Humanos , Opistorquiasis/diagnóstico , Opistorquiasis/epidemiología , Opistorquiasis/parasitología , Fosfatidilinositoles/metabolismoRESUMEN
BACKGROUND: Patients infected with a parasite often develop opisthorchiasis viverrini, which often progresses into cholangiocarcinoma (CCA) due to the asymptomatic nature of the infection. Currently, there are no effective diagnostic methods for opisthorchiasis or cholangiocarcinoma. OBJECTIVE: The aim of this study was to identify the host-responsive protein that can be developed as a diagnostic biomarker of opisthorchiasis and cholangiocarcinoma. METHODS: Plasma samples were collected from non-OVCCA, OV, and CCA subjects, and the proteomes were investigated by LC-MS/MS. Venn diagrams and protein network prediction by STITCH were used to identify the potential biomarkers. The level of candidate protein, the plasma checkpoint protein 1 (Chk1), was measured by indirect enzyme-linked immunosorbent assay (ELISA). RESULTS: Chk1 was present in the center of the protein network analysis in both the OV and CCA groups. In addition, the plasma Chk1 levels were significantly increased in both groups (P< 0.05). The sensitivity of the opisthorchiasis viverrini and cholangiocarcinoma was 59.38% and 65.62%, respectively, while the specificity of both was 85.71%. CONCLUSION: Chk1 was identified by differential plasma proteomes and was increased in O. viverrini-infected and cholangiocarcinoma-derived plasma samples. Higher levels of plasma Chk1 levels may serve as a potential diagnostic biomarker for opisthorchiasis and cholangiocarcinoma.
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Neoplasias de los Conductos Biliares , Colangiocarcinoma , Opistorquiasis , Opisthorchis , Animales , Neoplasias de los Conductos Biliares/patología , Conductos Biliares Intrahepáticos/metabolismo , Conductos Biliares Intrahepáticos/parasitología , Conductos Biliares Intrahepáticos/patología , Biomarcadores/metabolismo , Colangiocarcinoma/patología , Cromatografía Liquida , Humanos , Opistorquiasis/diagnóstico , Opistorquiasis/metabolismo , Opistorquiasis/parasitología , Espectrometría de Masas en TándemRESUMEN
Opisthorchis viverrini is a fish-borne zoonotic trematode that causes significant public health problems in Southeast Asia. Its life cycle requires Bithynia snails as the first intermediate hosts, fish, and human and/or carnivore hosts. This study assessed impacts of land use practice for rice cultivation and seasonality on the transmission dynamics of O. viverrini in Bithynia siamensis goniomphalos in rice paddy field habitats. The transmission of O. viverrini cercariae in B. s. goniomphalos was monitored at monthly intervals over a 4-year period from January 2010 to December 2013. From a total of 59,727 snails examined by standard cercarial shedding, the prevalence of O. viverrini was 0.7% (range, 0.0-4.1%). The prevalence of O. viverrini infection in B. s. goniomphalos varied with the amount of rainfall, with peaks of infection occurring in the cool-dry season, that is, after each rainy season. A shift of peak prevalence from cool-dry to hot-dry season observed in 2013 was associated with the increase in preceding water irrigation to support the production of second annual rice crop. Significant positive correlations were found between the prevalence and intensity of cercarial infection and the size of snails. Our results revealed substantial variation between years so that to have a clear understanding of the population dynamics of this complex system, studies should be conducted over an extended period (> 1 year). Results from this study highlight that water irrigation schemes in rice paddy cultivation and seasonality have a significant effect on the prevalence of O. viverrini in B. s. goniomphalos.
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Riego Agrícola , Opistorquiasis/transmisión , Opisthorchis/fisiología , Caracoles/parasitología , Riego Agrícola/métodos , Animales , Cercarias/fisiología , Producción de Cultivos , Humanos , Opistorquiasis/epidemiología , Opistorquiasis/parasitología , Oryza , Prevalencia , Lluvia , Estaciones del Año , Tailandia/epidemiologíaRESUMEN
Opisthorchis viverrini infection, opisthorchiasis, is a food-borne trematodiasis that is the main cause of cholangiocarcinoma, a bile duct cancer, in the Lower Mekong sub-region of Lao PDR, Cambodia, Vietnam, and Thailand. Despite extensive research on opisthorchiasis, the eradication of this disease has yet to be achieved. One of the major reasons for this failure is due to the multi-host life cycle of the parasite, which requires complex medical and public health interventions to eradicate. Another reason is due to a lack of knowledge of not only the interactions between the parasite and the human immune system, but also the interactions between the parasite and its various hosts during its complicated life cycle. Recent advances in various high-throughput omics technologies has allowed for the identification of key biomolecules crucial to the processes of parasitic transmission, and the identification of novel drug and/or vaccine targets. In this paper, omics studies dealing with O. viverrini host-parasite biology will be reviewed. In particular, there will be a focus on the strategies O. viverrini uses to trigger, evade, and manipulate the host's defense systems. Recently-identified biological molecules with potential as targets for interventions will also be reviewed. The results obtained from these omics approaches to analyzing O. viverrini and host interactions will be of great importance in the future when developing effective and sustainable medical and public health models for the prevention and control of opisthorchiasis and opisthorchiasis-induced CCA.
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Neoplasias de los Conductos Biliares/etiología , Neoplasias de los Conductos Biliares/prevención & control , Colangiocarcinoma/etiología , Colangiocarcinoma/prevención & control , Genómica , Metabolómica , Opistorquiasis/prevención & control , Proteómica , Animales , Humanos , Opistorquiasis/parasitologíaRESUMEN
The snail Bithynia siamensis goniomphalos acts as the first intermediate host of the human liver fluke, Opisthorchis viverrini, a major cause of cholangiocarcinoma in Northeast Thailand. In this study, we investigated host-parasite interactions in wet- and dry-season rice paddy plantation. The snails that aestivated through 4-month-dried period after rice harvest had average mortality rate of 36.45% and monthly increasing from 17.76% to 54.57%. Surprisingly, at depths greater than 5 cm experienced a higher mortality rate than upper level. Average mortality rate at the depth more than 5 cm, was 42.97%. In the initial month of aestivation, mortality rate of 0-5 and 5-10 cm depth was 9.13% and 26.39% then increase to 57.58% and 51.97%, respectively in the last 4th month. The average prevalence O. viverrini infection in snails during the study period was 0.44%. The highest prevalence of O. viverrini infection was found in the cool dry and wet season. The odds of O. viverrini infection in female snails and large snails were higher relative to male snails and medium-sized snails. The physicochemical factors involved in increased prevalence of O. viverrini infection in snail hosts were mean daily air temperature and average monthly rainfall. Short aestivation period made lower mortality in irrigation area than the previous report of non-irrigation area, that is, the aestivation is one of snail population control.
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Agricultura , Estivación/fisiología , Interacciones Huésped-Parásitos/fisiología , Opistorquiasis/veterinaria , Estaciones del Año , Caracoles/parasitología , Animales , Femenino , Masculino , Oryza , Temperatura , TailandiaRESUMEN
The multifunctional calreticulin (CALR) was identified as a major calcium-binding protein of the endoplasmic reticulum before being recognized as a chaperone in the same place. Only later were activities of calreticulin outside the endoplasmic reticulum described that for example affect cell proliferation and the innate immune system. In the present work we have investigated those extracellular activities of CALR from the cancerogenic human liver fluke Opisthorchis viverrini (OvCALR), as they might be important in host/parasite interaction. We first demonstrate that OvCALR is released from the parasite and stimulates a specific humoral immune response. Recombinant OvCALR is then shown to suppress proliferation of primary endothelial cells, their motility and sprouting activities. The potential of OvCALR to interfere with the complement system is established, firstly by demonstrating its direct binding to C1q and, secondly by suppression of hemolysis of sensitized red blood cells. These findings suggest that OvCALR is an important parasite antigen that could modulate diverse host functions and support parasite survival.
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Antígenos Helmínticos/metabolismo , Calreticulina/metabolismo , Complemento C1q/metabolismo , Interacciones Huésped-Parásitos , Células Endoteliales de la Vena Umbilical Humana/citología , Opisthorchis/metabolismo , Animales , Antígenos Helmínticos/farmacología , Calreticulina/farmacología , Movimiento Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Complemento C1q/efectos de los fármacos , Cricetinae , Células Endoteliales/citología , Células Endoteliales/efectos de los fármacos , Hemólisis/efectos de los fármacos , Células Endoteliales de la Vena Umbilical Humana/efectos de los fármacos , Humanos , Ratones , Neovascularización Fisiológica , OpistorquiasisRESUMEN
There have been considerable advances in our understanding of the systematics and ecology of Opisthorchis viverrini; however, this new knowledge has not only clarified but also complicated the situation. We now know that what was once considered to be a single species is, in fact, a species complex, with the individual species being confined to specific wetland areas. There is also a strong genetic association between the members of the O. viverrini species complex and their Bithynia snail intermediate hosts. Although this does not negate data collected before the recognition of this situation, it does lead to the caveat that regional and temporal variations in data collected may be related to the species examined. The advances in ecology have generally been spatially limited and have led, in part, to contradictory results that may well be related to nonrecognition of the species studied. It may also be related to natural temporal and spatial variation related, for example, to habitat characteristics. To understand the variation present, it will be necessary to conduct long-term (several years at least) sampling projects after defining the genetic characteristics of O. viverrini sensu lato and its Bithynia snail intermediate hosts.
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Clasificación , Ecología , Genética de Población , Opisthorchis/clasificación , Distribución Animal , Animales , Marcadores Genéticos/genética , Interacciones Huésped-Parásitos , Humanos , Estadios del Ciclo de Vida , Opisthorchis/genéticaRESUMEN
Four isoforms of calcium binding proteins containing 2 EF hand motifs and a dynein light chain-like domain in the human liver fluke Opisthorchis viverrini, namely OvCaBP1, 2, 3, and 4, were characterized. They had molecular weights of 22.7, 21.6, 23.7, and 22.5 kDa, respectively and showed 37.2-42.1% sequence identity to CaBP22.8 of O. viverrini. All were detected in 2- and 4-week-old immature and mature parasites. Additionally, OvCaBP4 was found in newly excysted juveniles. Polyclonal antibodies against each isoform were generated to detect the native proteins in parasite extracts by Western blot analysis. All OvCaBPs were detected in soluble and insoluble crude worm extracts and in the excretory-secretory product, at approximate sizes of 21-23 kDa. The ion-binding properties of the proteins were analyzed by mobility shift assays with the divalent cations Ca2+, Mg2+, Zn2+, and Cu2+. All OvCaBPs showed mobility shifts with Ca2+ and Zn2+. OvCaBP1 showed also positive results with Mg2+ and Cu2+. As tegumental proteins, OvCaBP1, 2, and 3 are interesting drug targets for the treatment of opisthorchiasis.
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Proteínas de Unión al Calcio/química , Motivos EF Hand , Proteínas del Helminto/química , Opistorquiasis/genética , Opisthorchis/parasitología , Animales , Anticuerpos Antihelmínticos , Calcio/metabolismo , Proteínas de Unión al Calcio/inmunología , Proteínas del Helminto/inmunología , Peso Molecular , Isoformas de ProteínasRESUMEN
Objective: To investigate the maturity development of miracidia in uterine eggs from four portions of the Opisthorchis viverrini uterus and environmental factors possibly affected in maturation and infectivity of distal part uterine eggs. Methods: Uteri of adult worms were divided into 4 equal parts. Development of eggs was determined under light microscope. Only embryonated eggs were used to evaluate the effects of physicochemical factors: temperature, salinity, acidity, ultraviolet A, B, C. Infection success was evaluated by feeding treated eggs to intermediate host snails and determining by using a PCR approach. Results: Eggs obtained from the uterus closest to the ovary (regions 1 and 2) failed to develop in vitro. Eggs from region 4 of the uterus (close to the genital pore) were used to study effects of physicochemical factors. The highest survival and infection success was in groups of eggs kept at 30 ℃ (95.20%). The calculated period of loss infection success (LI50 and LI95) on miracidia in distal uterine eggs by exposure to UV-A, UV-B and UV-C were 73 and 1523 d; 8 and 20 d; 1 and 2 d, respectively. Lethal concentrations (LC50 and LC95) of salinity, HCl and NaOH on miracidia in distal uterine eggs were 45.43 and 120.09 ppt, 0.01 and 0.25 M; 0.01 and 0.11 M, respectively, after 24 h exposure. Conclusions: Opisthorchis viverrini eggs display a high tolerance to environmental conditions, especially after snail host eating for infection.
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Opisthorchis viverrini, a carcinogenic fish borne fluke, requires freshwater snails and fish as intermediate hosts. Opisthorchiasis is endemic in parts of Southeast Asia, including Central and South Vietnam. In this region the transmission by intermediate hosts has received little attention. Therefore, freshwater snails and wild fish from Bau My Tho, an opisthorchiasis endemic area in Binh Dinh Province were collected for examination of O. viverrini cercariae and metacercariae, respectively. A total of 12,000 snails belonging to six families, of which 1616 Bithynia snails representing Bithynia siamensis goniomphalos and Bithynia funiculata; as well as 754 fish representing 12 species were examined. Shedding of O. viverrini cercariae was observed only in B. s. goniomphalos and B. funiculata, at infection rates of 0.86% and 0.14%, respectively. O. viverrini infection in Bithynia spp. was significantly associated with the habitat but not with the species and the shell size of Bithynia spp. O. viverrini metacercariae were found in 10 fish species representing both Cyprinidae and non-Cyprinidae families. The prevalence of O. viverrini infection in fish was significantly associated with species. Carassius auratus, a fish species commonly eaten raw, Rasbora aurotaenia and Puntius brevis had the highest prevalence of 74.0%, 55.8% and 31.6%, respectively. Sharing of the same snail and fish intermediate host species was found for O. viverrini and a O. viverrini duck-genotype that are sympatric in the study region. This study is the first to report on the intermediate host species of O. viverrini in Central Vietnam and indicates a high risk of acquiring opistorchiasis when eating raw fish dishes.
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Cercarias/parasitología , Peces/parasitología , Opistorquiasis/veterinaria , Caracoles/parasitología , Animales , Genotipo , Opistorquiasis/epidemiología , Prevalencia , Tailandia/epidemiología , Vietnam/epidemiologíaRESUMEN
OBJECTIVE: To explore larvicidal effects of anthelmintic drugs on Opisthorchis viverrini (O. viverrini) for alternative approach to interrupting its cycle for developing a field-based control program. METHODS: The larvicidal activities of albendazole (Al), artesunate (Ar), praziquantel (Pzq) and miltefosine (Mf) on O. viverrini cercariae and mature metacercariae were investigated. Lethal concentrations (LC50 and LC95) of these drugs were determined. Mature metacercariae previously exposed to various concentrations of the drugs were administered to hamsters. Worms were harvested 30 d post infection and worm recovery rates calculated. Al, Ar, Pzq and Mf produced morphological degeneration and induced shedding tails of cercariae after 24 h exposure. RESULTS: The LC50 and LC95 of Al, Ar, Pzq and Mf on cercariae were 0.720 and 1.139, 0.350 and 0.861, 0.017 and 0.693, and 0.530 and 1.134 ppm, respectively. LC50 and LC95 of Ar on mature metacercariae were 303.643 and 446.237 ppm and of Mf were 289.711 and 631.781 ppm, respectively but no lethal effect in Pzq- and Al-treated groups (up to 1 ppt). No worms were found in hamsters administered Pzq-treated metacercariae. The adult worms from Al-treated metacercariae were significantly bigger in size compared to the control group (P < 0.05). Fecundity and body width were greater in adults from Mf-treated metacercariae compared to the control group (P < 0.05). CONCLUSIONS: The larvicidal effects of these drugs were high efficacy to O. viverrini cercariae but lesser efficacy to metacercariae. It should be further studied with the eventual aim of developing a field-based control program.
RESUMEN
Calreticulin (CALR), a multifunctional protein thoroughly researched in mammals, comprises N-, P-, and C-domain and has roles in calcium homeostasis, chaperoning, clearance of apoptotic cells, cell adhesion, and also angiogenesis. In this study, the spatial and temporal expression patterns of the Opisthorchis viverrini CALR gene were analyzed, and calcium-binding and chaperoning properties of recombinant O. viverrini CALR (OvCALR) investigated. OvCALR mRNA was detected from the newly excysted juvenile to the mature parasite by RT-PCR while specific antibodies showed a wide distribution of the protein. OvCALR was localized in tegumental cell bodies, testes, ovary, eggs, Mehlis' gland, prostate gland, and vitelline cells of the mature parasite. Recombinant OvCALR showed an in vitro suppressive effect on the thermal aggregation of citrate synthase. The recombinant OvCALR C-domain showed a mobility shift in native gel electrophoresis in the presence of calcium. The results imply that OvCALR has comparable function to the mammalian homolog as a calcium-binding molecular chaperone. Inferred from the observed strong immunostaining of the reproductive tissues, OvCALR should be important for reproduction and might be an interesting target to disrupt parasite fecundity. Transacetylase activity of OvCALR as reported for calreticulin of Haemonchus contortus could not be observed.
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Calreticulina/genética , Calreticulina/metabolismo , Expresión Génica , Opisthorchis/genética , Opisthorchis/metabolismo , Animales , Calcio/metabolismo , Calreticulina/fisiología , Citrato (si)-Sintasa/metabolismo , Fertilidad/genética , Técnicas In Vitro , Chaperonas Moleculares , Opisthorchis/fisiología , Proteínas Recombinantes , Reproducción/genética , Distribución TisularRESUMEN
Bithynia siamensis goniomphalos is a freshwater snail that serves as the first intermediate host of the human liver fluke Opisthorchis viverrini. This parasite is a major public health problem in different countries throughout the Greater Mekong sub-region (Thailand, southern Vietnam, Lao PDR and Cambodia). Chronic O. viverrini infection also results in a gradual increase of fibrotic tissues in the biliary tract that are associated with hepatobiliary diseases and contribute to cholangiocarcinoma (a fatal type of bile duct cancer). Infectivity of the parasite in the snail host is strongly correlated with destruction of helminths by the snail's innate immune system, composed of cellular (hemocyte) and humoral (plasma) defense factors. To better understand this important host-parasite interface we applied sequential window acquisition of all theoretical spectra mass spectrometry (SWATH-MS) to identify and quantify the proteins from the hemolymph of B. siamensis goniomphalos experimentally infected with O. viverrini and compare them to non-infected snails (control group). A total of 362 and 242 proteins were identified in the hemocytes and plasma, respectively. Of these, 145 and 117 proteins exhibited significant differences in expression upon fluke infection in hemocytes and plasma, respectively. Among the proteins with significantly different expression patterns, we found proteins related to immune response (up-regulated in both hemocyte and plasma of infected snails) and proteins belonging to the structural and motor group (mostly down-regulated in hemocytes but up-regulated in plasma of infected snails). The proteins identified and quantified in this work will provide important information for the understanding of the factors involved in snail defense against O. viverrini and might facilitate the development of new strategies to control O. viverrini infection in endemic areas.
Asunto(s)
Opisthorchis/fisiología , Proteínas/genética , Caracoles/genética , Caracoles/parasitología , Animales , Hemolinfa/metabolismo , Hemolinfa/parasitología , Interacciones Huésped-Parásitos , Humanos , Opistorquiasis/parasitología , Opistorquiasis/transmisión , Opisthorchis/genética , Proteínas/metabolismo , Proteómica , Caracoles/metabolismoRESUMEN
Opisthorchiasis caused by Opisthorchis viverrini infection, is a serious public health problem in Lao PDR, Cambodia, Vietnam and Thailand. Cyprinid fish are required for the transmission of O. viverrini metacercariae to fish eating-mammal hosts including humans. The transmission of infective metacerariae is an important event, potentially resulting in the development of cholangiocarcinoma in cases of chronic opisthorchiasis. For a better understanding of the development of O. viverrini metacercariae in the second intermediate host, Barbonymus gonionotus (Cyprinidae), the immune response of infected B. gonionotus through fibrous tissue formation and superoxide dismutase (SOD) activity were investigated. The immunosuppressant (prednisolone) was administered to B. gonionotus infected with O. viverrini cercariae to study infectivity and development of the metacercariae. Additionally, the effects of prednisolone were studied by the investigation of fibrous tissue formation around the metacercariae, as well as SOD activity. In our study, prednisolone administered prior to infection in B. gonionotus had higher metacercarial recovery when compared to the non-administered control group. On the other hand, the administration of prednisolone after the O. viverrini infection allowed metacercariae to develop to the infective stage. Energy consumption from encystation to the infective stage required 443.9-463.7degree-days. The results show that immunosuppression decreases fibrous tissue and SOD activity in infected fish, and also affects the ability of O. viverrini metacercariae to develop to the infective stage. The immune response to metacercarial development of O. viverrini in cyprinid fish should be further studied to gain a greater understanding of this process, and hence, a better understanding of the mechanism(s) involved in metacercarial wall formation.