Changes in the Populations of Two Lymnaeidae and Their Infection by Fasciola hepatica and/or Calicophoron daubneyi over the Past 30 Years in Central France.

Field investigations were carried out during three periods (from 1976 to 1997, in 2013-2014, and in 2020-2021) on 39 cattle-raising farms on acidic soils to track changes in the populations of two Lymnaeidae (Galba truncatula and Omphiscola glabra) and their infection with Fasciola hepatica and/or Calicophoron daubneyi. Compared to the survey between 1976 and 1997 on these farms, there was a significant decrease in the number of the two lymnaeid populations and the size of the G. truncatula populations in both 2013-2014 and 2020-2021. This decline was significantly faster in the last nine years than it was before 2013. The area of habitats colonized by G. truncatula showed no significant variation over the years, while that of habitats with O. glabra significantly decreased in the period covered by the three surveys. The prevalence of F. hepatica infection in snails significantly decreased over the years, while C. daubneyi infection increased over time in both lymnaeid species. These changes are due to the use of triclabendazole to treat fasciolosis in ruminants since the 1990s, and are probably a consequence of the successive heatwaves that have occurred since 2018 in the region.

Fasciola hepatica: the dispersal of cercariae shed by the snail Galba truncatula.

Field investigations in 14 wild watercress beds located in the French region of Limousin, a known endemic area for distomatosis, were performed for three years to determine the distance that cercariae of Fasciola hepatica can reach in water before their encystment on the host plant. Each bed was located on the course of an open drainage furrow, while snails (Galba truncatula) lived upstream around the emergence of a source. Five plant species were collected in early April and examined to find metacercariae. Most cysts were noted on Nasturtium officinale (188 on 48.7 kg of dripped plants), followed by Helosciadium nodiflorum (125 on 33.4 kg). On the other plant species, there were few larvae. Most cercariae encysted on the plants growing in the most upstream part of each bed, usually on the first 50 cm in length. When water in the beds was fast running, the distribution of metacercariae was more limited and their number was fewer than those in the beds fed by a slow flow of water. Cercariae were able to swim or were carried away by the current up to a mean of 5 m in slow-flow waters before encysting; this distance was only 4 m in faster waters. Plants growing on the most upstream section of a watercress bed located in a drainage furrow are the most used by cercariae for their encystment, when snails live around the emergence of a source. The speed of the water current affected the number and distribution of metacercariae in the bed.

TITLE: Fasciola hepatica : la dispersion des cercaires émises par le mollusque Galba truncatula. ABSTRACT: Des investigations de terrain dans 14 cressonnières naturelles localisées dans la région française du Limousin, zone connue d'endémie de distomatose, ont été effectuées pendant trois années pour déterminer la distance que les cercaires de Fasciola hepatica peuvent atteindre dans l'eau avant leur enkystement sur la plante hôte. Chaque cressonnière était située sur le cours d'une rigole de drainage superficiel, tandis que les mollusques (Galba truncatula) vivaient plus en amont autour de l'émergence d'une source. Cinq espèces végétales ont été recueillies au début d'avril et examinées pour y trouver des métacercaires. La plupart des kystes ont été observés sur Nasturtium officinale (188 sur 48,7 kg de plantes égouttées), suivis par Helosciadium nodiflorum (125 sur 33,4 kg). Sur les autres espèces végétales, il y avait peu de larves. La plupart des cercaires étaient enkystées sur les plantes qui poussaient dans la partie la plus en amont de chaque cressonnière (habituellement sur les premiers 50 cm de longueur). Lorsque l'eau courante dans les cressonnières était rapide, la distribution des métacercaires était plus limitée et leur nombre était inférieur à celui observé dans les stations alimentées par un débit d'eau lent. Les cercaires ont été capables de nager ou ont été entrainées par le courant jusqu'à 5 m en moyenne dans une eau à écoulement lent avant leur enkystement ; cette distance n'était que de 4 m dans les eaux plus rapides. Les plantes qui poussent sur la section la plus en amont d'une cressonnière située dans une rigole de drainage sont les plus utilisées par les cercaires pour leur enkystement lorsque les mollusques vivent autour de l'émergence d'une source. La vitesse du courant d'eau affecte le nombre et la distribution des métacercaires dans la cressonnière.

Neopterin and CXCL-13 in Diagnosis and Follow-Up of Trypanosoma brucei gambiense Sleeping Sickness: Lessons from the Field in Angola.

Human African Trypanosomiasis may become manageable in the next decade with fexinidazole. However, currently stage diagnosis remains difficult to implement in the field and requires a lumbar puncture. Our study of an Angolan cohort of T. b. gambiense-infected patients used other staging criteria than those recommended by the WHO. We compared WHO criteria (cell count and parasite identification in the CSF) with two biomarkers (neopterin and CXCL-13) which have proven potential to diagnose disease stage or relapse. Biological, clinical, and neurological data were analysed from a cohort of 83 patients. A neopterin concentration below 15.5 nmol/L in the CSF denoted patients with stage 1 disease, and a concentration above 60.31 nmol/L characterized patients with advanced stage 2 (trypanosomes in CSF and/or cytorachia higher than 20 cells) disease. CXCL-13 levels below 91.208 pg/mL denoted patients with stage 1 disease, and levels of CXCL-13 above 395.45 pg/mL denoted patients with advanced stage 2 disease. Values between these cut-offs may represent patients with intermediate stage disease. Our work supports the existence of an intermediate stage in HAT, and CXCL-13 and neopterin levels may help to characterize it.

Current decline in schistosome and soil-transmitted helminth infections among school children at Loum, Littoral region, Cameroon.

INTRODUCTION: Soil-transmitted helminth infections (STHs) and schistosomiasis have serious consequences for the health, education and nutrition of children in developing countries. As Loum is known as a highly endemic commune for these infections, several deworming campaigns have been carried out in the past. The purpose of this study was to determine any changes that have occurred since then in the characteristics of these infections among schoolchildren in this site. METHODS: A cross-sectional study was conducted in October 2016 on 289 schoolchildren. Stool and urine samples were collected and examined to determine the prevalence and intensity of helminth infections. RESULTS: The highest prevalence was noted for Schistosoma haematobium (34.2%), followed by Ascaris lumbricoides (8.6%), S. mansoni (4.9%) and Trichuris trichiura (4.9%) in decreasing order. A prevalence of less than 2% was noted for each of the other two helminths. The highest mean intensity was found for S. haematobium (39.6 eggs/10 ml of urine), followed by A. lumbricoides (24.2 eggs per gram of faeces: epg), Strongyloides stercoralis (16.6 epg) and Schistosoma mansoni (12.3 epg). The prevalence of T. trichiura was significantly higher in boys and that of S. haematobium in children aged 10 years or older, while the differences between other values of prevalence or between egg burdens were not significant. CONCLUSION: Compared with values reported in 2003, the prevalence and intensity of schistosomiases and STH infections in Loum has sharply decreased in 2016. Confirmation of this decrease in the years to come allowed to space deworming campaigns among schoolchildren.

Proteome characterization in various biological fluids of Trypanosoma brucei gambiense-infected subjects.

Human African trypanosomiasis (HAT) is a neglected tropical disease that is endemic in sub-Saharan Africa. Control of the disease has been recently improved by better screening and treatment strategies, and the disease is on the WHO list of possible elimination. However, some physiopathological aspects of the disease transmission and progression remain unclear. We propose a new proteomic approach to identify new targets and thus possible new biomarkers of the disease. We also focused our attention on fluids classically associated with HAT (serum and cerebrospinal fluid (CSF)) and on the more easily accessible biological fluids urine and saliva. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) established the proteomic profile of patients with early and late stage disease. The serum, CSF, urine and saliva of 3 uninfected controls, 3 early stage patients and 4 late stage patients were analyzed. Among proteins identified, in CSF, urine and saliva, respectively, 37, 8 and 24 proteins were differentially expressed and showed particular interest with regards to their function. The most promising proteins (Neogenin, Neuroserpin, secretogranin 2 in CSF; moesin in urine and intelectin 2 in saliva) were quantified by enzyme-linked immunosorbent assay in a confirmatory cohort of 14 uninfected controls, 23 patients with early stage disease and 43 patients with late stage disease. The potential of two proteins, neuroserpin and moesin, with the latter present in urine, were further characterized. Our results showed the potential of proteomic analysis to discover new biomarkers and provide the basis of the establishment of a new proteomic catalogue applied to HAT-infected subjects and controls. SIGNIFICANCE: Sleeping sickness, also called Human African Trypanosomiasis (HAT), is a parasitic infection caused by a parasitic protozoan, Trypanosoma brucei gambiense or T. b. rhodesiense which are transmitted via an infected tsetse fly: Glossina. For both, the haemolymphatic stage (or first stage) signs and symptoms are intermittent fever, lymphadenopathy, hepatosplenomegaly, headaches, pruritus, and for T. b. rhodesiense infection a chancre is often formed at the bite site. Meningoencephalitic stage (or second stage) occurs when parasites invade the CNS, it is characterised by neurological signs and symptoms such as altered gait, tremors, neuropathy, somnolence which can lead to coma and death if untreated. first stage of the disease is characterizing by fevers, headaches, itchiness, and joint pains and progressive lethargy corresponding to the second stage with confusion, poor coordination, numbness and trouble sleeping. Actually, diagnosing HAT requires specialized expertise and significant resources such as well-equipped health centers and qualified staff. Such resources are lacking in many endemic areas that are often in rural locales, so many individuals with HAT die before the diagnosis is established. In this study, we analysed by mass spectrometry the entire proteome of serum, CSF, urine and saliva samples from infected and non-infected Angolan individuals to define new biomarkers of the disease. This work of proteomics analysis is a preliminary stage to the characterization of the whole proteome, of these 4 biological fluids, of HAT patients. We have identified 69 new biomarkers. Five of them have been thoroughly investigated by ELISA quantification. Neuroserpine and Moesin are respectively promising new biomarkers in CSF and urine's patient for a better diagnosis.

Diversity of Toxoplasma gondii strains shaped by commensal communities of small mammals.

Commensal rodent species are key reservoirs for Toxoplasma gondii in the domestic environment. In rodents, different T. gondii strains show variable patterns of virulence according to host species. Toxoplasma gondii strains causing non-lethal chronic infections in local hosts will be more likely to persist in a given environment, but few studies have addressed the possible role of these interactions in shaping the T. gondii population structure. In addition, the absence of validated techniques for upstream detection of T. gondii chronic infection in wild rodents hinders exploration of this issue under natural conditions. In this study, we took advantage of an extensive survey of commensal small mammals in three coastal localities of Senegal, with a species assemblage constituted of both native African species and invasive species. We tested 828 individuals for T. gondii chronic infection using the modified agglutination test for antibody detection in serum samples and a quantitative PCR assay for detection of T. gondii DNA in brain samples. The infecting T. gondii strains were genotyped whenever possible by the analysis of 15 microsatellite markers. We found (i) a very poor concordance between molecular detection and serology in the invasive house mouse, (ii) significantly different levels of prevalence by species and (iii) the autochthonous T. gondii Africa 1 lineage strains, which are lethal for laboratory mice, only in the native African species of commensal small mammals. Overall, this study highlights the need to reconsider the use of MAT serology in natural populations of house mice and provides the first known data about T. gondii genetic diversity in invasive and native species of small mammals from Africa. In light of these results, we discuss the role of invasive and native species, with their variable adaptations to different T. gondii strains, in shaping the spatial structure of T. gondii genetic diversity in Africa.

Local low dose curcumin treatment improves functional recovery and remyelination in a rat model of sciatic nerve crush through inhibition of oxidative stress.

Traumatic injuries to peripheral nerves are frequent, however, specific pharmacological treatments are currently lacking. Curcumin has antioxidant, anti-inflammatory and neuroprotective properties but high oral doses are required for therapeutic use, particularly due to its low bioavailability. The aim of the present study was to investigate the effects of local and continuous treatment using low curcumin doses on functional recovery and nerve regeneration after rat sciatic nerve crush (SNC). Curcumin was administered by osmotic pumps with a catheter delivering the drug at the injury site (0.2 mg/day for 4 weeks). Functionally, early improvements in mechanical sensitivity, finger spacing of the injured paw, skilful walking and grip strength were observed in curcumin-treated animals. The curcumin treatment increased expression of compact myelin proteins (MPZ and PMP22), myelin sheath thickness and, correspondingly, increased motor and sensitive nerve conduction velocity. Microscopic analysis of gastrocnemius muscle indicated a curcumin-induced decrease in neurogenic lesions. Curcumin treatment reduced the production of reactive oxygen species (ROS) (which were notably produced by macrophages), lipid peroxidation and increased expression of transcription factor Nrf2. In silico analyses indicated that curcumin combines all the characteristics required to be an efficient lipid peroxidation inhibitor at the heart of biological membranes, hence protecting their degradation due to ROS. This antioxidant capacity is likely to contribute to the beneficial effects of curcumin after SNC injury. These results demonstrate that, when administrated locally, low doses of curcumin represent a promising therapy for peripheral nerve regeneration.

Determination of zones at risk for fasciolosis in the department of Haute-Vienne, central France: a retrospective study on natural infections detected in 108,481 Galba truncatula for 37 years.

A retrospective study on the natural infection of Galba truncatula by Fasciola hepatica was carried out in the French department of Haute-Vienne to determine whether there are areas at risk for fasciolosis. Adult snails included in this analysis came from samples collected from pastures on 259 farms and from 121 wild watercress beds between 1970 and 2006. Fasciola hepatica infection rates were examined in relation to altitude and climatic data (mean annual rainfall, mean annual temperature) of each municipality. In a total of 108,481 snails collected in 151 municipalities, the overall prevalence of infection was 3.8% but varied according to the municipalities from which samples were taken (from 1% to 7.4%). The prevalence of F. hepatica infection in snails significantly decreased when the mean altitude of municipalities or their mean annual rainfall increased. However, this prevalence significantly increased with increasing mean annual temperatures. Studying the prevalence of infection in these snails makes it possible to delineate zones at risk for fasciolosis on the acid soils of Haute-Vienne. The risk of infection for livestock would be greater in areas of Haute-Vienne below 400 m above sea level and would gradually decrease when the altitude of the land increases.

TITLE: Détermination des zones à risque pour la fasciolose dans le département de la Haute-Vienne (France) : une étude rétrospective sur les infestations naturelles détectées chez 108.481 Galba truncatula pendant 37 années. ABSTRACT: Une étude rétrospective sur l'infestation naturelle de Galba truncatula par Fasciola hepatica a été effectuée dans le département de la Haute-Vienne pour déterminer s'il existe des zones à risque pour la fasciolose. Les limnées adultes impliquées dans cette analyse proviennent d'échantillons prélevés dans les pâturages de 259 fermes et dans 121 cressonnières sauvages entre 1970 et 2006. Les taux d'infection par Fasciola hepatica ont été mis en relation avec l'altitude et les données climatiques (précipitations annuelles moyennes, température annuelle moyenne) de chaque municipalité. Sur un total de 108.481 limnées récoltées dans 151 municipalités, la prévalence globale de l'infestation était de 3,8 %, mais variait selon les municipalités dans lesquelles les échantillons ont été prélevés (de 1 % à 7,4 %). La prévalence de l'infestation par F. hepatica diminue considérablement lorsque l'altitude moyenne des municipalités ou leurs précipitations annuelles moyennes augmente. D'autre part, cette prévalence augmente considérablement avec l'augmentation de la température annuelle moyenne des municipalités. L'étude de la prévalence de l'infestation chez ces limnées permet de délimiter des zones à risque pour la fasciolose sur les sols acides de la Haute-Vienne. Le risque d'infestation pour le bétail serait plus élevé dans les zones de la Haute-Vienne situées en dessous de 400 m d'altitude et diminuerait graduellement lorsque l'altitude de ces terrains augmente.

Toxoplasma gondii survey in cats from two environments of the city of Rio de Janeiro, Brazil by Modified Agglutination Test on sera and filter-paper.

BACKGROUND: Toxoplasma gondii is a protozoan with a worldwide distribution, in warm-blood animals, including humans. Local conditions and environmental disturbances may influence transmission dynamics of a zoonotic agent. This study evaluates the epidemiology of T. gondii based on toxoplasmosis prevalence in two populations of cats living in distinct urbanization conditions in Rio de Janeiro, Brazil. METHODS: Among 372 domestic cats sampled, 265 were from a public shelter located downtown Rio and 107 from a relatively preserved wild environment in a residential area. Sera and eluates from dried blood spots were tested for detection of IgG antibodies against T. gondii by modified agglutination test (MAT). RESULTS: Antibodies to T. gondii were detected in 32/265 (12.08%) animals from the public shelter and in 4/107 (3.74%) cats from the residential area. Identical results were observed for sera and eluates. CONCLUSIONS: Filter paper provides a reliable accurate alternative storage option when conditions of sample collection and transportation in the field are unfavorable. The significantly lower prevalence in the residential area is discussed in terms of environmental, biological and behavioral features.

Current decline in the number and size of Galba truncatula and Omphiscola glabra populations, intermediate hosts of Fasciola hepatica, on the acidic soils of Central France.

Field investigations on the habitats colonized by Galba truncatula or Omphiscola glabra were carried out on 162 farms of the Limousin region, Central France, to determine whether there is currently a decline in the number and size of snail populations. Seven types of snail habitats were considered here. Compared to the numbers of snail populations recorded from 1976 to 1992, the values noted from 2013 to 2016 were significantly lower, with a decline rate of 34% for G. truncatula and 23% for O. glabra. Variations in this decline rate with the type of snail habitat were also noted. The greatest decreases in the numbers of snail populations were noted for spring heads located in meadows and for road ditches, while the lowest were noted for open drainage furrows present in meadows. The distribution of these habitats according to their area did not show any significant change over time. In contrast, overwintering snails were significantly less numerous in 2013-2016 in five types of habitats for G. truncatula and in three types only for O. glabra. Several causes underlie this population decline. Among them, the current development of mechanical cleaning in open drainage systems and road ditches, that of subsurface drainage in meadows, and regular gyro-crushing of vegetation around temporary spring heads were the most important.

Larval trematode infections in Lymnaea glabra populations living in the Brenne Regional Natural Park, central France.

Lymnaea glabra is known to be a natural intermediate host of two flukes, Calicophoron daubneyi and Fasciola hepatica, in central France. But it can also sustain larval development of other digeneans. Adult snails were thus collected from 206 habitats in 2014 and 2015 to identify parasite species and determine the prevalence of each digenean infection in relation to the five types of snail habitats. Seven digenean species were noted in 321 infected snails (out of 17,647 L. glabra). Snails with F. hepatica or C. daubneyi were found in 14.5% and 12.6% of habitats, respectively. Percentages were lower for snails with Opisthoglyphe ranae (5.8%), Haplometra cylindracea (5.3%) and were less than 5% for those infected with Echinostoma revolutum, Notocotylus sp. or Plagiorchis sp. Prevalence noted for each parasite species varied with the type of habitat. The number of species in L. glabra was lower than that found in G. truncatula from the same region (7 instead of 10). The distribution and prevalence of each digenean species were thus dependent on the type and location of each snail habitat.

Interferon gamma effect on immune mediator production in human nerve cells infected by two strains of Toxoplasma gondii.

Interferon gamma (IFN-γ) is the major immune mediator that prevents toxoplasmic encephalitis in murine models. The lack of IFN-γ secretion causes reactivation of latent T. gondii infection that may confer a risk for severe toxoplasmic encephalitis. We analyse the effect of IFN-γ on immune mediator production and parasite multiplication in human nerve cells infected by tachyzoites of two T. gondii strains (RH and PRU). IFN-γ decreased the synthesis of MCP-1, G-CSF, GM-CSF and Serpin E1 in all cell types. It decreased IL-6, migration inhibitory factor (MIF) and GROα synthesis only in endothelial cells, while it increased sICAM and Serpin E1 synthesis only in neurons. The PRU strain burden increased in all nerve cells and in contrast, RH strain replication was controlled in IFN-γ-stimulated microglial and endothelial cells but not in IFN-γ-stimulated neurons. The proliferation of the PRU strain in all stimulated cells could be a specific effect of this strain on the host cell.

Fasciola hepatica: comparative metacercarial productions in experimentally-infected Galba truncatula and Pseudosuccinea columella.

As large numbers of metacercariae of Fasciola hepatica are necessary for research, experimental infections of Galba truncatula and Pseudosuccinea columella with this digenean were carried out to determine the better intermediate host for metacercarial production and, consequently, the most profitable snail for decreasing the cost price of these larvae. Pre-adult snails (4 mm in shell height) originating from two populations per lymnaeid species were individually exposed to two or five miracidia, raised at 23 °C and followed for cercarial shedding up to their death. Compared to values noted in G. truncatula, the survival of P. columella on day 30 post-exposure was significantly greater, while the prevalence of F. hepatica infection was significantly lower. In the four P. columella groups, metacercarial production was significantly greater than that noted in the four groups of G. truncatula (347-453 per cercariae-shedding snail versus 163-275, respectively). Apart from one population of G. truncatula, the use of five miracidia per snail at exposure significantly increased the prevalence of F. hepatica in P. columella and the other population of G. truncatula, whereas it did not have any clear effect on the mean number of metacercariae. The use of P. columella for experimental infections with F. hepatica resulted in significantly higher metacercarial production than that noted with G. truncatula, in spite of a lower prevalence for the former lymnaeid. This finding allows for a significant decrease in the cost price of these larvae for commercial production.

Detection of Toxoplasma gondii DNA in horse meat from supermarkets in France and performance evaluation of two serological tests.

In France, some cases of severe toxoplasmosis have been linked to the consumption of horse meat that had been imported from the American continent where atypical strains of Toxoplasma gondii are more common than in Europe. Many seroprevalence studies are presented in the literature but risk assessment of T. gondii infection after horse meat consumption is not possible in the absence of validated serological tests and the unknown correlation between detection of antibodies against T. gondii and presence of tissue cysts. We performed magnetic-capture polymerase chain reaction (MC-PCR) to detect T. gondii DNA in 231 horse meat samples purchased in supermarkets in France and evaluated the performance and level of agreement of the modified agglutination test (MAT) and enzyme-linked immunosorbent assay (ELISA) in the meat juices. The serological tests lacked sensitivity, specificity, and agreement between them, and there was no correlation with the presence of T. gondii DNA in horse meat, raising concerns about the reliability of T. gondii seroprevalence data in horses from the literature. T. gondii DNA was detected in 43% of horse meat samples but the absence of strain isolation in mice following inoculation of more than 100 horse meat samples suggests a low distribution of cysts in skeletal muscles and a low risk of T. gondii infection associated with horse meat consumption. However, to avoid any risk of toxoplasmosis, thorough cooking of horse meat is recommended.

Pseudosuccinea columella: age resistance to Calicophoron daubneyi infection in two snail populations.

Individual infections of Egyptian and French Pseudosuccinea columella with five miracidia of Calicophoron daubneyi were carried out to determine whether this lymnaeid was capable of sustaining larval development of this parasite. On day 42 post-exposure (at 23 °C), infected snails were only noted in groups of individuals measuring 1 or 2 mm in height at miracidial exposure. Snail survival in the 2-mm groups was significantly higher than that noted in the 1-mm snails, whatever the geographic origin of snail population. In contrast, prevalence of C. daubneyi infection was significantly greater in the 1-mm groups (15-20% versus 3.4-4.0% in the 2-mm snails). Low values were noted for the mean shell growth of infected snails at their death (3.1-4.0 mm) and the mean number of cercariae (<9 in the 1-mm groups, <19 in the 2-mm snails). No significant differences between snail populations and snails groups were noted for these last two parameters. Most infected snails died after a single cercarial shedding wave. Both populations of P. columella showed an age resistance to C. daubneyi infection and only juveniles measuring 2 mm or less in shell height at exposure can ensure larval development of this digenean up to cercarial shedding.

Ruminal paramphistomosis in cattle from northeastern Algeria: prevalence, parasite burdens and species identification.

Slaughterhouse samples were analysed over a two-year period (September 2010-August 2012) in Jijel (northeastern Algeria) in order to determine seasonal variations in the prevalence and intensity of bovine paramphistomosis in a Mediterranean climate and identify paramphistome species using molecular biology. In spring and summer, significantly higher prevalences and lower parasite burdens were noted in bull calves, thus indicating an effect of season on these parameters. In contrast, the differences among seasonal prevalences or among seasonal parasite burdens were not significant in the case of old cows. Eleven adult worms from the slaughterhouses of Jijel and three neighbouring departments (Constantine, El Tarf and Setif) were analysed using molecular markers for species identification. Two different species, Calicophoron daubneyi and C. microbothrium, were found. The presence of these two paramphistomids raises the question of their respective frequency in the definitive host and local intermediate hosts.

Adaptation of Lymnaea fuscus and Radix balthica to Fasciola hepatica through the experimental infection of several successive snail generations.

BACKGROUND: High prevalence of Fasciola hepatica infection (>70%) was noted during several outbreaks before the 2000s in several French farms where Galba truncatula is lacking. Other lymnaeids such as Lymnaea fuscus, L. glabra and/or Radix balthica are living in meadows around these farms but only juvenile snails can sustain complete larval development of F. hepatica while older snails were resistant. The low prevalence of infection (<20%) and limited cercarial production (<50 cercariae per infected snail) noted with these juveniles could not explain the high values noted in these cattle herds. As paramphistomosis due to Calicophoron daubneyi was not still noted in these farms, the existence of another mode of infection was hypothesized. Experimental infection of several successive generations of L. glabra, originating from eggs laid by their parents already infected with this parasite resulted in a progressive increase in prevalence of snail infection and the number of shed cercariae. The aim of this paper was to determine if this mode of snail infection was specific to L. glabra, or it might occur in other lymnaeid species such as L. fuscus and R. balthica. METHODS: Five successive generations of L. fuscus and R. balthica were subjected to individual bimiracidial infections in the laboratory. Resulting rediae and cercariae in the first four generations were counted after snail dissection at day 50 p.e. (20°C), while the dynamics of cercarial shedding was followed in the F5 generation. RESULTS: In the first experiment, prevalence and intensity of F. hepatica infection in snails progressively increased from the F1 (R. balthica) or F2 (L. fuscus) generation. In the second experiment, the prevalence of F. hepatica infection and the number of shed cercariae were significantly lower in L. fuscus and R. balthica (without significant differences between both lymnaeids) than in G. truncatula. CONCLUSION: The F. hepatica infection of several successive snail generations, coming from parents infected with this parasite, resulted in a progressive increase in prevalence and intensity of snail infection. This may explain high prevalence of fasciolosis noted in several cattle-breeding farms when the common snail host of this digenean, G. truncatula, is lacking.

In vitro infection of human nervous cells by two strains of Toxoplasma gondii: a kinetic analysis of immune mediators and parasite multiplication.

The severity of toxoplasmic infection depends mainly on the immune status of the host, but also on the Toxoplasma gondii strains, which differ by their virulence profile. The relationship between the human host and T. gondii has not yet been elucidated because few studies have been conducted on human models. The immune mechanisms involved in the persistence of T. gondii in the brains of immunocompetent subjects and during the reactivation of latent infections are still unclear. In this study, we analyzed the kinetics of immune mediators in human nervous cells in vitro, infected with two strains of T. gondii. Human neuroblast cell line (SH SY5Y), microglial (CMH5) and endothelial cells (Hbmec) were infected separately by RH (type I) or PRU (type II) strains for 8 h, 14 h, 24 h and 48 h (ratio 1 cell: 2 tachyzoites). Pro-inflammatory protein expression was different between the two strains and among different human nervous cells. The cytokines IL-6, IL-8 and the chemokines MCP-1 and GROα, and SERPIN E1 were significantly increased in CMH5 and SH SY5Y at 24 h pi. At this point of infection, the parasite burden declined in microglial cells and neurons, but remained high in endothelial cells. This differential effect on the early parasite multiplication may be correlated with a higher production of immune mediators by neurons and microglial cells compared to endothelial cells. Regarding strain differences, PRU strain, but not RH strain, stimulates all cells to produce pro-inflammatory growth factors, G-CSF and GM-CSF. These proteins could increase the inflammatory effect of this type II strain. These results suggest that the different protein expression profiles depend on the parasitic strain and on the human nervous cell type, and that this could be at the origin of diverse brain lesions caused by T. gondii.

Lymnaea cubensis, an experimental intermediate host for Fascioloides magna.

Single-miracidium infections of Lymnaea cubensis (Pfeiffer) from Guadeloupe with the giant liver fluke Fascioloides magna (Bassi, 1875) (Digenea) were carried out during five successive snail generations to determine if this lymnaeid might sustain complete larval development of the parasite. Controls were constituted by a French population of Galba truncatula (Miller) (a single generation) infected according to the same protocol. It was recorded that prevalence and intensity of F. magna infection in L. cubensis progressively increased from F1 to F5 generations. Cercarial shedding of F. magna was noted only within F5 generation of L. cubensis. However, most measured parameters of infection in this species were significantly lower than those noted for G. truncatula and most L. cubensis died after a single shedding wave. Despite this, L. cubensis can be added to the list of potential intermediate hosts of F. magna.

Radix natalensis: the effect of Fasciola hepatica infection on the reproductive activity of the snail.

Experimental infections of Egyptian Radix natalensis (shell height at miracidial exposure: 4 mm) with a French isolate of Fasciola hepatica were carried out under laboratory conditions at 22 °C to specify the characteristics and follow the dynamics of their egg-laying. Controls constituted unexposed R. natalensis of the same size. No significant difference between controls and the uninfected snails of the exposed group was noted, whatever the parameter considered. In controls and exposed snails, the dates of the first egg masses were close to each other (56.4-65.3 days). In contrast, the life span of snails and the length of the egg-laying period were significantly shorter and egg production was significantly lower in infected R. natalensis than in controls and uninfected snails. In infected R. natalensis, but without cercarial shedding (NCS snails), egg production was irregular throughout the egg-laying period. In cercarial-shedding (CS) snails, the first egg masses were laid before the first cercarial emergence (at a mean of 56 days and 67 days, respectively). Thereafter, egg mass production of CS snails was irregular up to day 72 of the experiment, stopped during the following two weeks and started again after day 88 for a single snail. In conclusion, the F. hepatica infection of R. natalensis reduced the reproductive activity in both NCS and CS snails. The pattern noted for egg production in infected R. natalensis seems to be species-specific because of the high shell size of this lymnaeid and its role as an atypical intermediate host in the life cycle of the parasite.