[New eating habits, new parasitic risks: The example of fish]. / De nouvelles habitudes alimentaires, de nouveaux risques parasitaires : l'exemple du poisson.

INTRODUCTION: In the past decades, the massive explosion of "Japanese" restaurants serving raw fish popularised new culinary habits in France. At the same time, consumers have made a habit of preparing raw or pickled fish dishes themselves at home. As a result, the identification of live parasitic worm larvae in raw fish flesh is common and a source of concern for professionals or amateur cooks. Sometimes, these worms are spit out or removed after fibroscopy in patients developing severe epigastric pain quickly after eating raw fish. This paper is aiming at having a quick review of the main parasites transmitted to humans by eating raw fish in France. METHODS: This article is based on the personal experience of the authors, on references preferentially from the French literature and on the results of the Fish Parasites (ANR) research program. RESULTS: From 2011 to 2014, Fish-Parasites (ANR) assessed the prevalence of parasitism in sea and freshwater fish belonging to 29 species. About 57% of sea fish were parasitised by Anisakidae. Larvae of Dibothriocephalus latus were found in pike, perch, and burbot in Lake Geneva but in none of the fish examined from Annecy or Le Bourget lakes. Concerning human anisakidosis, a retrospective survey was carried out in the years 2010 to 2014 among all medical parasitology laboratories from university hospitals in France. Thirty-seven cases of anisakidosis have been reported, including 18 cases of allergic anisakidosis. Six additional cases of severe Anisakidae allergy were reported to the National Allergovigilance Network over the same period. CONCLUSIONS: Despite the increase in consumption of raw fish, and compared to previous studies, cases of anisakidosis are decreasing, but their allergenic potential is increasing. The incidence of dibothriocephalosis, after some trend of emergence on the shores of Lake Geneva some 20 years ago, is currently decreasing, but sporadic cases of importation are still reported. Actions with professionals (investigation, providing of information) and research programs on management of parasitic risk control are being pursued and have resulted in an update of the technical instruction of the French General Directorate for Food on the control of parasitic risk in fish.

Trichinella and polar bears: a limited risk for humans.

In this review, we identified 63 cases reported since World War II of human trichinellosis linked to the consumption of parasitized polar bear (Ursus maritimus) meat. This low number contrasts to the numerous cases of human trichinellosis related to consumption of the meat of black (U. americanus) or brown bears (U. arctos). The prevalence of Trichinella infection is high in bears, but larval muscular burden is usually lower in polar bears compared to other bear species. Polar bears, therefore, seem to play a limited role in the transmission of trichinellosis to humans, as native residents living in the Arctic traditionally consume well-cooked bear meat, and travellers and foreign hunters have only limited access to this protected species due to the declining polar bear population.

Confirmation and follow-up of neurocysticercosis by real-time PCR in cerebrospinal fluid samples of patients living in France.

Neurocysticercosis diagnosis is based on a combination of clinical, epidemiological, radiological, and immunological findings. We describe a real-time PCR assay for the confirmation of neurocysticercosis diagnosis in cerebrospinal fluid. The assay, tested on samples from nine patients living in France and diagnosed with neurocysticercosis, had a detection rate of 83.3% and 100% specificity.

[Emergence of Candida parapsilosis candidemia at Cochin hospital. Characterization of isolates and search for risk factors]. / Émergence de candidémies à Candida parapsilosis à l'hôpital Cochin. Caractérisation des isolats et recherche de facteurs de risque.

Candida parapsilosis is a normal saprophyte of the skin, characterized by their affinity for catheters. This species has, in vitro, a level of sensitivity against the echinocandins, significantly lower than that observed with other Candida species. Recently, new species: Candida orthopsilosis and Candida metapsilosis, phenotypically identical to C. parapsilosis, have been identified by molecular biology. From 2003 to 2007, in the Cochin hospital, the proportion of C. parapsilosis among non-albicans species isolated from blood cultures increased from 17 (3/18) to 38% (5/13). To understand the reasons for this emergence, we retrospectively characterized isolates, conducted a case-control and researched a link between the emergence and introduction of caspofungin in our hospital. We analysed data from 26 patients who had candidemia with C. parapsilosis. Genotypic analysis of isolates has not identified the new species C. orthopsilosis and C. metapsilosis. The case-control study showed a broad-spectrum antibiotics was significantly more frequent for candidemia with C. parapsilosis compared to C. albicans (52 versus 26%, P=0.04) as a previous treatment with caspofungin (11 versus 0%, P=0.04). The introduction of caspofungin is contemporary with the emergence of candidemia with C. parapsilosis with a tendency to be related to its level of consumption in the ICU. Our results should encourage biologists to closely monitor the frequency and level of sensitivity of strains of C. parapsilosis isolated in hospital.

[Diagnosis of invasive pulmonary aspergillosis: value of bronchoalveolar lavage galactomannan for immunocompromised patients]. / Diagnostic de l'aspergillose bronchopulmonaire invasive: valeur du dosage du galactomannane dans le lavage bronchoalvéolaire de patients immunodéprimés.

Invasive pulmonary aspergillosis (IPA) is an emerging disease associated with high mortality. The diagnosis is difficult, based on a combination of elements that are clinical, radiological and biological. For early detection of cases of IPA, during 25 months, we have systematically carried out on the LBA (N=355) of immunocompromised patients (N=313) a determination of Aspergillus galactomannan (GM) by ELISA (PlateliaAspergillus, BioRad). We observed 14 cases of probable API. The sensitivity of GM compared to direct examination (DE) and culture is, respectively, 64% versus 29% and 57%. The determination of GM is definitely more sensitive than the ED. Excellent specificity (98%) allows its implementation as a screening test in patients at risk.

Factors of occurrence of ocular toxoplasmosis. A review.

Acquired and congenital toxoplasmosis are frequently complicated by ocular toxoplasmosis. The diagnosis relies on clinical aspects, response to specific treatment and results of biological assays. The incidence and the prevalence of this complication are difficult to establish precisely and depend on the prevalence of the parasite infection in the general population, and are affected by factors such as type of exposure to the parasite, genetic backgrounds of the parasite and the host, and type of immune response elicited by the parasite.

Contributions of immunoblotting, real-time PCR, and the Goldmann-Witmer coefficient to diagnosis of atypical toxoplasmic retinochoroiditis.

Ocular toxoplasmosis is a major cause of posterior uveitis worldwide. The diagnosis is based mainly on ophthalmological examination. Biological diagnosis is necessary in atypical cases, and this requires aqueous humor sampling by anterior chamber paracentesis. We evaluated real-time PCR targeting the Toxoplasma gondii 529-bp repeat element, the Goldmann-Witmer coefficient (GWC), and immunoblotting for the diagnosis of toxoplasmic retinochoroiditis in 54 patients with atypical uveitis. The results of these biological tests, applied to paired aqueous humor-serum samples, were compared to the clinical findings. Combining either PCR or the GWC with immunoblotting increased the sensitivity to 73% or 70%, respectively. Together, PCR and the GWC had 80% sensitivity. If feasible, sensitivity can be increased by combining the three methods (85% sensitivity). The interval between symptom onset and anterior chamber paracentesis strongly influenced the detection of specific intraocular antibody synthesis. The sensitivity of the GWC increased from 45% to 56% when sampling was performed 10 days after symptom onset, and that of immunoblotting increased from 53% to 72% when puncture was performed 30 days after symptom onset. PCR analysis of aqueous humor samples detected toxoplasmic DNA in 55% of patients. In contrast to the results of immunoblotting and the GWC, the results of PCR were not influenced by the interval between symptom onset and paracentesis. PCR was more informative than the GWC and immunoblotting for immunocompromised patients. Acute necrotizing retinal lesions were significantly larger in PCR-positive patients, with a mean of 3.5 optic disc diameters, than in PCR-negative patients, with a mean of 1.5 optic disc diameters.

Trichinellosis acquired in Senegal from warthog ham, March 2009.

Three confirmed and three suspected cases of trichinellosis have been reported in France with onset of symptoms in March 2009, linked to consumption of smoked warthog ham in Senegal.

Trichinellosis acquired in Nunavut, Canada in September 2009: meat from grizzly bear suspected.

Five cases of trichinellosis with onset of symptoms in September 2009, were reported in France, and were probably linked to the consumption of meat from a grizzly bear in Cambridge Bay in Nunavut, Canada. Travellers should be aware of the risks of eating raw or rare meat products in arctic regions, particularly game meat such as bear or walrus meat.

[How can the cost of screening for toxoplasmosis during pregnancy be reduced?]. / Comment réduire le coût du dépistage de la toxoplasmose chez la femme enceinte?

BACKGROUND: A program of systematic serology screening for toxoplasmosis during pregnancy has been running in France since 1978. The program involves monthly follow-ups for all non-immune pregnant women. Due to the steady decline in the seroprevalence of toxoplasmosis, the cost of the program is steadily increasing. Current screening is based on the detection of IgG and IgM isotypes. The aim of this work was to estimate the benefit of replacing combined dosage of two isotypes, by an alternative strategy that detects total anti-Toxoplasma immunoglobulins. METHODS: The rate of decreasing seroprevalence and the increasing burden on serological examinations was measured in a study population of pregnant women who were checked for toxoplasmosis by the parasitology laboratory of the Cochin Hospital, Paris. The increase in screening costs was estimated for the all-pregnant women and the expected benefits stemming from simply measuring total anti-Toxoplasma immunoglobulins compared to the double IgG-IgM assay were estimated. RESULTS: Between 1987 and 2008, the seroprevalence of toxoplasmosis measured at the Cochin hospital dropped from 70.8% to 48.6% with a 1.77% annual rate of decline. This downward trend is similar to that observed by the national perinatal surveys performed in 1995 and in 2003. As the number of non-immune women to follow-up each month is constantly increasing, the proportion of negative tests issued reached 87.6% in 2008. Extrapolating these results to the whole of France, we estimated that the number of required screening tests perform was increasing by 93,000 units per year with an additional associated cost of one million euros. Various alternative scenarios of antibody detection are proposed that could save between 40.2% and 48.4% of current screening costs. CONCLUSION: Replacement of combined dosage of IgG and IgM isotypes by determination of just total Ig would significantly reduce costs of toxoplasmosis screening for pregnant women, without effecting either the general strategy, or proven efficiency of the national program.

Use of nuclear and mitochondrial DNA PCR and sequencing for molecular identification of Diphyllobothrium isolates potentially infective for humans.

Tapeworms of the genus Diphyllobothrium (Cobold, 1858) are widely distributed all around the world and some of them are agents of human diphyllobothriasis. Approximately 50 species have been described within the Diphyllobothrium genus but only 13 are human pathogens. Species identification by using morphological criteria is very difficult. We determined the value of 18S ribosomal RNA gene, internal transcribed spacer (ITS) and cytochrome c oxidase subunit 1 gene (COI) sequences to differentiate between Diphyllobothrium isolates. Sequences from 18 isolates (larvae or adults) of D. latum, D. nihonkaiense, D. ditremum, D. dentriticum and D. stemmacephalum species were obtained. COI region sequences analysis was clearly more discriminative than those of the ITS1 and 18S rRNA and was a useful tool for identifying specimens.

[Troponin dosage in a patient with asymptomatic myocarditis due to trichinellosis]. / Myocardite asymptomatique au cours d''un cas de trichinellose: intérêt du dosage de la troponine.

Human trichinellosis is a potentially severe parasitic disease occurring after ingestion of undercooked meat infected with Trichinella sp. larvae. We report the case of a patient who ate an undercooked bear meat hunted in Canada; he presented with the usual symptoms of trichinellosis (i.e, facial oedema, myalgias and fever) complicated with an asymptomatic myocarditis. Myocarditis is a rare, but potentially lethal complication of trichinellosis. Myocarditis should be screened systematically even when specific symptoms are missing; dosage of troponin serum is a simple and reliable mean for such screening.

A novel Toxoplasma gondii calcium-dependent protein kinase.

Toxoplasma gondii is an obligate intracellular parasite that infects all types of cells in humans. A family of calcium-dependent protein kinases (CDPKs), previously identified as important in the development of plants and protists, was recently shown to play a role in the infectivity of apicomplexans, and in motility and host cell invasion in particular. We report here the isolation of a new calcium-dependent protein kinase gene from the human toxoplasmosis parasite, Toxoplasma gondii. The gene consists of 12 exons. The encoded protein, TgCDPK4, consists of the four characteristic domains of members of the CDPK family and is most similar to PfCDPK2 from Plasmodium falciparum. We measured TgCDPK4 activity, induced by calcium influx, using a kinase assay. A calcium chelator (EGTA) inhibited this activity. These findings provide evidence of signal transduction involving members of the CDPK family in T. gondii.

[First report of a human case of trichinellosis due to Trichinella britovi after jackal (Canis aureus) meat consumption in Algeria]. / Première observation d'un cas humain de trichi- nellose due à Trichinella britovi en Algérie après consommation de viande de chacal (Canis aureus).

We report a single case of trichinellosis contracted in Algeria (Batna region), in a practising Moslim. Shortly after returning to France in November 2004, the patient developed the typical clinical and biological signs of the disease. Although the patient claimed having only eaten mutton, an unusual host for Trichinella, a meticulous investigation revealed that he also had eaten a grilled leg of jackal (Canis aureus). One of the four Trichinella larvae detected in a muscular biopsy enabled us to identify the parasite as Trichinella britovi by a multiplex PCR analysis. This is the first identification of the etiological agent of sylvatic trichinellosis occurring in North Africa and the first case of symptomatic trichinellosis due to jackal meat consumption in Africa.

Human trichinellosis due to Trichinella britovi in southern France after consumption of frozen wild boar meat.

Six patients were infected with Trichinella britovi in southern France following consumption of frozen wild boar meat, which had been frozen at -35 degrees C for 7 days. Microscopic examination of a sample of frozen wild boar muscle revealed the presence of rare encapsulated Trichinella larvae, identified as T. britovi. People eating wild boar must follow individual prophylactic rules such as efficient cooking of meat (at least 65 degrees C at the core for 1 minute) as recommended by the International Commission on Trichinellosis, or freezing exceeding four weeks at -20 degrees C.

[Prevalence of Diphyllobothrium latum, L., 1758 infestation in Perca fluviatilis from Lake Leman]. / Prévalence de l'infestation par Diphyllobothrium latum, L., 1758 chez les Perches (Perca fluvatilas) du lac Léman.

Diphyllobothriasis is contracted by consuming raw or undercooked freshwater fish and is still present on the shores of lake Leman. The aim of this study was to evaluate the prevalence of Diphyllobothrium latum plerocercoid larvae in Perca fluviatilis from this lake. Four to 10% of perch fillets examined in November 2003, February 2004, April 2004 and January 2005, were infested with D. latum larvae. The identification of the larvae was confirmed by PCR and sequencing of the 18S rDNA.

A multifocal outbreak of trichinellosis linked to horse meat imported from North America to France in 1993.

An outbreak of 538 cases of trichinellosis occurred in France in December 1993. Seven cases developed neurotrichinosis and 23 had cardiologic complications. No deaths were recorded. Two patients had a positive muscle biopsy showing living Trichinella larvae. One of them was typed as Trichinella spiralis. A case-control study showed that horse meat was the only meat associated with illness (odds ratio = 80.7). The risk of illness increased with the amount of horse meat eaten and when it was consumed raw. The cases, which were spread out in five foci, bought horse meat from five butchers who had received parts of a single horse carcass imported in November 1993 from Canada. The Trichinella International Screening Program, implemented since 1985 after two similar episodes involving a thousand cases, failed to detect the incriminated horse carcass. This new horse meat-related outbreak led to modifications of the internationally recommended screening methods whereby the weight of meat samples tested was increased.

Localized oral Fusarium infection in an AIDS patient with malignant lymphoma.

We report here a case of localized oral Fusarium infection in an AIDS patient who developed an ulceration in the soft palate. Fusarium solani was identified by histopathology and culture. We believe this to be the second reported case of oral Fusarium infection in a patient with haematological malignancy and the first reported association of oral Fusarium infection with AIDS.

Trichinellosis: a worldwide zoonosis.

Trichinella spp. are some of the most widespread parasites infecting people and other mammals all over the world, regardless of climate. This paper attempts to describe the present status of trichinellosis worldwide and to determine if and why trichinellosis is emerging or re-emerging. The global prevalence of the disease is difficult to evaluate but as many as 11 million people may be infected. More than 10000 cases of human trichinellosis were reported by the International Commission on Trichinellosis from 1995 to June 1997 and about 10000 porcine infections were reported by the Office International des Epizooties in 1998. The disease is particularly worrisome in the Balkans, Russia, the Baltic republics, in some parts of China and Argentina. Horsemeat-related outbreaks have been reported in France and Italy and have involved about 3000 patients in the past 25 years. The emergence of trichinellosis in some countries is explained by a better knowledge of the disease (formerly often misdiagnosed as influenza), modifications of consumer habits, re-forestation in Europe and increase of wild game, importation of meats from countries where trichinellosis is endemic and failure of veterinary control due to human error or to social upheavals. This disease linked to meat-consumption which is theoretically easy to prevent by adequate cooking, freezing and veterinary controls, should deserve the attention of all persons involved in public health and it could be eradicated at least from domestic pigs.

Experimental trichinellosis in horses: biological and parasitological evaluation.

Three groups of three horses each were, respectively, infected with 5000, 20,000 and 50,000 larvae of Trichinella spiralis. The strain used was isolated from a human biopsy during horsemeat-related outbreaks of trichinellosis in France. Transient muscular disorders were only observed in two of the horses infected with 50,000 larvae but none of the horses had fever. A significant increase in blood eosinophils was noticed in 5 horses. Serum LDH, aldolase and CPK peaked at the fifth week post-infection. Specific IgG assayed by indirect immunofluorescence and ELISA, appeared 2-5 weeks post-infection and disappeared between 16 and 40 weeks. The distribution of T. spiralis larvae was maximal in the tongue, masseters and diaphragm, but a large decrease in the number of larvae recovered from the muscles was noticed among the horses slaughtered at the beginning and end of the experiment. In muscular histological sections, larvae were observed in an intramyofibrillar position and were surrounded by a mild to severe inflammatory reaction.