Search

Oocysts of the protozoan Toxoplasma gondii are found in felid feces and can be washed into coastal waters, where they persist for months, attaching to algae and accumulating in invertebrates. We used wild bivalves to assess contamination of coastal waters of the Kerguelen and Galapagos archipelagos by this zoonotic parasite. Additionally, we leveraged the contrasting situations of these archipelagos to identify some potential drivers of contamination. In the Galapagos, with a cat density reaching 142 per km 2 , 15.38% of the sampled oysters (Saccostrea palmula) tested positive for T. gondii by quantitative real-time PCR (qPCR) (n = 260), and positive samples were found in all eight sampling sites. In Kerguelen, with 1-3 cats per km 2 , 40.83% of 120 tested mussels (Mytilus edulis platensis) were positive, and positive samples were found in four out of the five sampling sites. These findings provide evidence of T. gondii contamination in the coastal waters of these archipelagos. Furthermore, T. gondii-positive bivalves were found on islands located 20 km away (Galapagos) and 5 km away (Kerguelen) from the nearest cat population, indicating that T. gondii oocysts can disperse through waterborne mechanisms over several kilometers from their initial deposition site. In the Galapagos, where runoff is infrequent and all sites are exposed to currents, the prevalence of qPCR-positive bivalves did not show significant variations between sites (p = 0.107). In Kerguelen where runoff is frequent and site exposure variable, the prevalence varied significantly (p < 0.001). The detection of T. gondii in Kerguelen mussels was significantly correlated with the site exposure to currents (odds ratio (OR) 60.2, p < 0.001) and the on-site density of giant kelp forests (OR 2.624, p < 0.001). This suggests that bivalves can be contaminated not only by oocysts transported by currents but also by consuming marine aggregates containing oocysts that tend to form in kelp forests., (Copyright © 2024 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Aims: The protozoan parasites Cryptosporidium spp., Giardia duodenalis and Toxoplasma gondii are identified as public health priorities and are present in a wide variety of environments including the marine ecosystem. The objective of this study was to demonstrate that the marine bivalve blue mussel (Mytilus edulis) can be used as a tool to monitor the contamination of marine waters by the three protozoa over time., Methods and Results: In order to achieve a proof of concept, mussels were exposed to three concentrations of G. duodenalis cysts and Cryptosporidium parvum/T. gondii oocysts for 21 days, followed by 21 days of depuration in clear water. Then, natural contamination by these protozoa was sought for in wild marine blue mussels along the northwest coast of France to validate their relevance as bioindicators in the field. Our results highlighted that: (a) blue mussels bioaccumulated the parasites for 21 days, according to the conditions of exposure, and parasites could still be detected during the depuration period (until 21 days); (b) the percentage of protozoa-positive M. edulis varied under the degree of protozoan contamination in water; (c) mussel samples from eight out of nine in situ sites were positive for at least one of the protozoa., Conclusions: The blue mussel M. edulis can bioaccumulate protozoan parasites over long time periods, according to the degree of contamination of waters they are inhabiting, and can highlight recent but also past contaminations (at least 21 days)., Significance and Impact of the Study: Mytilus edulis is a relevant bioaccumulators of protozoan (oo)cysts in laboratory and field conditions, hence its potential use for monitoring parasite contamination in marine waters., (© 2021 The Society for Applied Microbiology.)

Aims: The study was aimed to understand the depuration process of Cryptosporidium parvum and Toxoplasma gondii oocysts by zebra mussel (Dreissena polymorpha), to consider the use of the zebra mussel as a bioremediation tool., Materials and Methods: Two experiments were performed: (i) individual exposure of mussel to investigate oocyst transfers between bivalves and water and (ii) in vivo exposure to assess the ability of the zebra mussel to degrade oocysts., Results: (i) Our results highlighted a transfer of oocysts from the mussels to the water after 3 and 7 days of depuration; however, some oocysts were still bioaccumulated in mussel tissue. (ii) Between 7 days of exposure at 1000 or 10 000 oocysts/mussel/day and 7 days of depuration, the number of bioaccumulated oocysts did not vary but the number of infectious oocysts decreased., Conclusion: Results show that D. polymorpha can release oocysts in water via (pseudo)faeces in depuration period. Oocysts remain bioaccumulated and infectious oocyst number decreases during the depuration period in zebra mussel tissues. Results suggest a degradation of bioaccumulated C. parvum and T. gondii oocysts., Significance and Impact of the Study: This study highlighted the potential use of D. polymorpha as a bioremediation tool to mitigate of protozoan contamination in water resources., (© 2020 The Society for Applied Microbiology.)

Nowadays, it is necessary to improve the efficiency of wastewater treatment plant treatments. In this context the use of biofilter species, like Dreissena polymorpha, as a bioremediation tool in wastewater is increasingly highlighted. The innovative aim of this study is to evaluate the zebra mussel survival in the outlet channel of a conventional WWTP to use them as bioremediation tool. For this, mussels were transplanted in the outlet channel for 28 days and different biomarkers were monitored. D. polymorpha is able to maintain itself in good physiological conditions until 21 days, yet at 28 days a high mortality rate (24%), a decrease in filtration efficiency (8/15 mussels filtered and 17.0% of filtration rate) and antioxidant system activation (CAT activity et gpx gene expression increase) suggest an exhaustion. Some biomarkers suggested a hypoxic stress. Despite the unfavourable conditions, bivalves have bioaccumulated pathogenic protozoa (Toxoplasma gondii and Giardia duodenalis) during the exposure. Zebra mussel seems to be a promising tool for bioremediation in wastewater., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

This study aimed to combine cellular and molecular analyses for better detail the effects of various stresses on a sentinel species of freshwater invertebrate. For this purpose, the hemocytes of the zebra mussel, Dreissena polymorpha, were exposed to different stresses at two different intensities, high or low: chemical (cadmium and ionomycin), physical (ultraviolet B), or biological ones (Cryptosporidium parvum and Toxoplasma gondii). After exposure, flow cytometry and droplet digital PCR analyses were performed on the same pools of hemocytes. Several responses related to necrosis, apoptosis, phagocytosis, production of nitric oxide and expression level of several genes related to the antioxidant, detoxification and immune systems were evaluated. Results showed that hemocyte integrity was compromised by both chemical and physical stress, and cellular markers of phagocytosis reacted to ionomycin and protozoa. While cadmium induced oxidative stress and necrosis, ionomycin tends to modulate the immune response of hemocytes. Although both biological stresses led to a similar immune response, C. parvum oocysts induced more effects than T. gondii, notably through the expression of effector caspases gene and an increase in hemocyte necrosis. This suggests different management of the two protozoa by the cell. This work provides new knowledge of biomarkers in the zebra mussel, at both cellular and molecular levels, and contributes to elucidate the mechanisms of action of different kinds of stress in this species., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Cryptosporidium parvum, Toxoplasma gondii and Giardia duodenalis are worldwide pathogenic protozoa recognized as major causal agents of waterborne disease outbreaks. To overcome the normative process (ISO 15553/2006) limitations of protozoa detection in aquatic systems, we propose to use the zebra mussel (Dreissena polymorpha), a freshwater bivalve mollusc, as a tool for biomonitoring protozoan contamination. Mussels were exposed to three concentrations of C. parvum oocysts, G. duodenalis cysts or T. gondii oocysts for 21 days followed by 21 days of depuration in clear water. D. polymorpha accumulated protozoa in its tissues and haemolymph. Concerning T. gondii and G. duodenalis, the percentage of protozoa positive mussels reflected the contamination level in water bodies. As for C. parvum detection, oocysts did accumulate in mussel tissues and haemolymph, but in small quantities, and the limit of detection was high (between 50 and 100 oocysts). Low levels of T. gondii (1-5 oocysts/mussel) and G. duodenalis (less than 1 cyst/mussel) were quantified in D. polymorpha tissues. The ability of zebra mussels to reflect contamination by the three protozoa for weeks after the contamination event makes them a good integrative matrix for the biomonitoring of aquatic ecosystems., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Toxoplasma gondii is a ubiquitous foodborne protozoan that can infect humans at low dose and displays different prevalences among countries in the world. Ingestion of food or water contaminated with small amounts of T. gondii oocysts may result in human infection. However, there are no regulations for monitoring oocysts in food, mainly because of a lack of standardized methods to detect them. The objectives of this study were (i) to develop a reliable method, applicable in biomonitoring, for the rapid detection of infectious oocysts by cell culture of their sporocysts combined with quantitative PCR (sporocyst-CC-qPCR) and (ii) to adapt this method to blue and zebra mussels experimentally contaminated by oocysts with the objective to use these organisms as sentinels of aquatic environments. Combining mechanical treatment and bead beating leads to the release of 84% ± 14% of free sporocysts. The sporocyst-CC-qPCR detected fewer than ten infectious oocysts in water within 4 days (1 day of contact and 3 days of cell culture) compared to detection after 4 weeks by mouse bioassay. For both mussel matrices, oocysts were prepurified using a 30% Percoll gradient and treated with sodium hypochlorite before cell culture of their sporocysts. This assay was able to detect as few as ten infective oocysts. This sporocyst-based CC-qPCR appears to be a good alternative to mouse bioassay for monitoring infectious T. gondii oocysts directly in water and also using biological sentinel mussel species. This method offers a new perspective to assess the environmental risk for human health associated with this parasite. IMPORTANCE The ubiquitous protozoan Toxoplasma gondii is the subject of renewed interest due to the spread of oocysts in water and food causing endemic and epidemic outbreaks of toxoplasmosis in humans and animals worldwide. Displaying a sensitivity close to animal models, cell culture represents a real alternative to assess the infectivity of oocysts in water and in biological sentinel mussels. This method opens interesting perspectives for evaluating human exposure to infectious T. gondii oocysts in the environment, where oocyst amounts are considered to be very small., (Copyright © 2019 American Society for Microbiology.)

The protozoa Toxoplasma gondii and Cryptosporidium parvum are public health priorities because their oocysts can persist in recreational, surface, drinking, river, and sea water sources for a long time. To evaluate the capacity of the freshwater crustacean Gammarus fossarum to accumulate T. gondii and C. parvum oocysts, gammarids were exposed to 200, 2000 or 20,000 oocysts per gammarid and per day for 21 days followed by 5 days of depuration. C. parvum DNA was detected by qPCR in G. fossarum in only one out of four pools for the highest concentration and after 14 days of exposure, and T. gondii DNA was detected after 7 days of exposure to the two highest concentrations. Our results document the capacity of G. fossarum to accumulate T. gondii in its tissues proportionally to the ambient concentration; the maximum number of oocysts was detected in gammarid tissues after exposure to 20,000 oocysts per day. Mean values of 3.26 (±3), 21.71 (±15.18), and 17.41 (±10.89) oocysts were detected in gammarids after 7, 14, and 21 days, respectively, and after 5 days of depuration, T. gondii oocysts were still present in gammarid tissues. These results show for the first time that a freshwater crustacean can bioaccumulate T. gondii oocysts, suggesting that G. fossarum is a potential effective bioindicator of protozoan contamination in biomonitoring studies. Moreover, due to its key position in freshwater food webs, G. fossarum could also play a role in the trophic transfer of protozoa., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Aims: The objective of this study was to evaluate if freshwater bivalves can be used to detect the presence of Toxoplasma gondii in water bodies., Methods and Results: Zebra mussels (Dreissena polymorpha) were caged for 1 month upstream and downstream of the discharge points of wastewater treatment plants (WWTPs). Physiological status was assessed to assure good health of bivalves during transplantation. The presence of T. gondii was investigated in mussel tissues by qPCR. In autumn, T. gondii was detected in mussels caged downstream of the discharge points of two WWTPs. In spring, it was detected upstream of one WWTP., Conclusions: For the first time, T. gondii DNA has been shown in a continental mollusc in environmental conditions. This highlights the interest of an active approach that could be applied independently of the presence or accessibility of autochthonous populations, and underlines the presence of T. gondii in natural waters under pressure of WWTP discharge at a certain time of the year., Significance and Impact of the Study: This study shows that transplanted zebra mussels could be used as biosamplers to reveal contamination of freshwater systems by T. gondii., (© 2015 The Society for Applied Microbiology.)

Water quality is a public health concern that calls for relevant biomonitoring programs. Molecular tools such as polymerase chain reaction (PCR) are progressively becoming more sensitive and more specific than conventional techniques to detect pathogens in environmental samples such as water and organisms. The zebra mussel (Dreissena polymorpha) has already been demonstrated to accumulate and concentrate various human waterborne pathogens. In this study, first, a spiking experiment to evaluate detection levels of Toxoplasma gondii DNA in zebra mussel organs using real-time PCR was conducted. Overall, lower DNA levels in the hemolymph, digestive gland, and remaining tissues (gonad and foot) were detected compared to mantle, muscle, and gills. Second, an in vivo experiment with 1000 T. gondii oocysts per mussel and per day for 21 consecutive days, followed by 14 days of depuration time in protozoa-free water was performed. T. gondii DNA was detected in all organs, but greatest concentrations were observed in hemolymph and mantle tissues compared to the others organs at the end of the depuration period. These results suggest that (i) the zebra mussel is a potential new tool for measuring T. gondii concentrations and (ii) real-time PCR is a suitable method for pathogen detection in complex matrices such as tissues.