Survival and infectivity of Toxoplasma gondii and Cryptosporidium parvum oocysts bioaccumulated by Dreissena polymorpha.

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.

Toxoplasma gondii infection in pork produced in France.

The aim of this study was to assess the seroprevalence of the Toxoplasma gondii parasite in pork produced in France, and to determine infection risk factors. An innovative survey was designed based on annual numbers of slaughtered pigs from intensive and outdoor farms in France. A total of 1549 samples of cardiac fluids were collected from pig hearts to determine seroprevalence using a Modified Agglutination Test. Of those, 160 hearts were bio-assayed in mice to isolate live parasites. The overall seroprevalence among fattening pigs was 2·9%. The adjusted seroprevalence in pigs from intensive farms was 3·0%; the highest in sows (13·4%); 2·9% in fattening pigs and 2·6% in piglets. Adjusted seroprevalence in fattening animals from outdoor farms was 6·3%. Strains were isolated from 41 animals and all were genotyped by Restriction Fragment Length Polymorphism as type II. Risk-factor analysis showed that the risk of infection was more than three times higher for outdoor pigs, and that sows' risk was almost five times higher than that of fattening animals. This study provides further evidence of extensive pork infection with T. gondii regardless of breeding systems, indicating that farm conditions are still insufficient to guarantee 'Toxoplasma-free pork'.

Zebra mussel as a new tool to show evidence of freshwater contamination by waterborne Toxoplasma gondii.

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.

The relationship between socioeconomic indices and potentially zoonotic pathogens carried by wild Norway rats: a survey in Rhône, France (2010-2012).

Leptospira interrogans, hantaviruses (particularly Seoul virus), hepatitis E virus (HEV), and Toxoplasma gondii are rat-associated zoonoses that are responsible for human morbidity and mortality worldwide. This study aimed to describe the infection patterns of these four pathogens in wild rats (Rattus norvegicus) across socioeconomic levels in neighbourhoods in Lyon, France. The infection or exposure status was determined using polymerase chain reaction or serology for 178 wild rats captured in 23 locations; additionally, confirmatory culture or mouse inoculation was performed. Multivariate logistic regression analyses were used to investigate whether morphological and socioeconomic data could predict the infection status of the rats. This study revealed that the rat colony's age structure may influence the prevalence of L. interrogans, hantavirus, and HEV. In addition, areas with high human population densities and low incomes may be associated with a greater number of infected rats and an increased risk of disease transmission.

In the rat liver, Adenoviral gene transfer efficiency is comparable to AAV.

Adenoviral (AdV) and Adenovirus-associated viral (AAV) vectors both are used for in vivo gene therapy of inherited liver disorders, such as Crigler-Najjar syndrome type 1. In a relevant animal model, the Gunn rat, both vectors efficiently correct the severe hyperbilirubinemia characteristic of this liver disorder. Although the clinical use of AAV is more advanced, as demonstrated by the successful phase 1 trial in hemophilia B patients, because of its large cloning capacity AdV remains an attractive option. A direct comparison of the efficacy of these two vectors in the liver in a relevant disease model has not been reported. Aim of this study was to compare the efficiency of clinically applicable doses of both vectors in the Gunn rat. AdV or scAAV (self-complimentary AAV) ferrying identical liver-specific expression cassettes of the therapeutic gene, UGT1A1, were injected into the tail vein. As the titration methods of these two vectors are very different, a comparison based on vector titers is not valid. Therefore, their efficacy was compared by determining the amount of vector genomes delivered to the liver required for therapeutic correction of serum bilirubin. Like AAV, the liver-specific first-generation AdV also provided sustained correction in this relevant disease model. UGT1A1 mRNA expression provided per genome was comparable for both vectors. Flanking the expression cassette in AdV with AAV-ITRs (inverted terminal repeats), increased UGT1A1 mRNA expression eightfold which resulted in a significant improvement of efficacy. Compared with AAV, less AdV genomes were needed for complete correction of hyperbilirubinemia.

Storm-induced transfer of particulate trace metals to the deep-sea in the Gulf of Lion (NW Mediterranean Sea).

In order to calculate budgets of particulate matter and sediment-bound contaminants leaving the continental shelf of the Gulf of Lion (GoL), settling particles were collected in March 2011 during a major storm, using sediment traps. The collecting devices were deployed in the Cap de Creus submarine canyon, which represents the main export route. Particulate matter samples were analyzed to obtain mass fluxes and contents in organic carbon, Al, Cr, Co, Ni, Cu, Zn, Cd, Pb and La, Nd and Sm. The natural or anthropogenic origin of trace metals was assessed using enrichment factors (EFs). Results are that Zn, Cu and Pb appeared to be of anthropogenic origin, whereas Ni, Co and Cr appeared to be strictly natural. The anthropogenic contribution of all elements (except Cd) was refined by acid-leaching (HCl 1 N) techniques, confirming that Zn, Cu and Pb are the elements that are the most enriched. However, although those elements are highly labile (59-77%), they do not reflect severe enrichment (EFs <4). Most particles originate from the Rhone River. This has been confirmed by two different tracing procedures using rare earth elements ratios and concentrations of acid-leaching residual trace metals. Our results hence indicate that even in this western extremity of the GoL, storm events mainly export Rhone-derived particles via the Cap de Creus submarine canyons to the deep-sea environments. This export of material is significant as it represents about a third of the annual PTM input from the Rhone River.

No tumour-initiating risk associated with scAAV transduction in newborn rat liver.

Delivery of recombinant adeno-associated virus (rAAV) vectors to the newborn liver is followed by a rapid loss of episomal vector copies because of hepatocyte proliferation. In selected hepatocytes, integration of rAAV genomes can lead to a sustained expression of the transgene. The safety of in vivo gene therapy with single-stranded AAV vectors has been questioned in a study reporting a high incidence of hepatocellular carcinoma, associated with provirus integration events in mice that receive an single-stranded AAV injection at birth. To investigate the tumour-initiating potential of the newly established self-complementary AAV (scAAV) vectors in the liver, groups of newborn rats received intravenous injection of a scAAV vector encoding the green fluorescent protein (GFP), or were injected with phosphate-buffered saline (PBS) or diethylnitrosamine (DEN), a well-known liver tumour initiator. The rats were fed on a diet containing 2-acetylaminofluorene, a potent liver tumour-promoting agent to accelerate the carcinogenic process. After 2 months, the animals were killed and their livers analysed. Preneoplastic nodules were identified by glutathion S-transferase-p (GSTp) staining, and GFP expression was detected by immunohistochemistry. Vector genome integration events were analysed. The numbers of GSTp-positive foci were comparable in the PBS and the scAAV-GFP groups and significantly higher in the DEN group. The proportion of GSTp-positive foci that also expressed GFP was low and in the range expected for random occurrence. No specific integration hot spots were detected by linear amplification-mediated-PCR in transduced liver. In conclusion, scAAV transduction of newborn rat liver does not trigger preneoplastic lesions suggesting an absence of liver tumourigenesis.

Induction of sulfadiazine resistance in vitro in Toxoplasma gondii.

We induced sulfadiazine resistance in two sulfadiazine sensitive strains of Toxoplasma gondii, RH (Type I) and ME-49 (Type II) in vitro by using drug pressure. At first, sulfadiazine susceptibility of the two sensitive strains and two naturally resistant strains of T. gondii was evaluated on Vero cells using an enzyme-linked immunosorbent assay (ELISA). The IC(50) values of sulfadiazine were 77 µg/mL for RH, 51 µg/mL for ME-49 and higher than 1000 µg/mL for the two natural resistant strains. Secondly, induced resistance of the strains by gradually increase sulfadiazine concentration was verified by this test, which resulted IC(50) values at higher than 1000 µg/mL. In conclusion we developed in vitro two sulfadiazine resistant strains called RH-R(SDZ) and ME-49-R(SDZ). These strains resistant to sulfadiazine would be useful to characterize resistance mechanisms to sulfadiazine.

Meiotic instability of human minisatellite CEB1 in yeast requires DNA double-strand breaks.

Minisatellites are tandemly repeated DNA sequences of 10-100-bp units. Some minisatellite loci are highly unstable in the human germ line, and structural analysis of mutant alleles has suggested that repeat instability results from a recombination-based process. To provide insights into the molecular mechanism of human minisatellite instability, we developed Saccharomyces cerevisiae strains carrying alleles of the most unstable human minisatellite locus, CEB1 (ref. 2). We observed that CEB1 is destabilized in meiosis, resulting in a variety of intra- and inter-allelic gains or losses of repeat units, similar to rearrangements described in humans. Using mutations affecting the initiation of recombination (spo11) or mismatch repair (msh2 pms1 ), we demonstrate that meiotic destabilization depends on the initiation of homologous recombination at nearby DNA double-strand break (DSBs) sites and involves a 'rearranged heteroduplex' intermediate. Most of the human and yeast data can be explained and unified in the context of DSB repair models.

Measuring particle size distribution and mass concentration of nanoplastics and microplastics: addressing some analytical challenges in the sub-micron size range.

HYPOTHESIS: The implementation of the proposal from the European Chemical Agency (ECHA) to restrict the use of nanoplastics (NP) and microplastics (MP) in consumer products will require reliable methods to perform size and mass-based concentration measurements. Analytical challenges arise at the nanometre to micrometre interface, e.g., 800 nm-10 µm, where techniques applicable at the nanometre scale reach their upper limit of applicability and approaches applicable at the micrometre scale must be pushed to their lower limits of detection. EXPERIMENTS: Herein, we compared the performances of nine analytical techniques by measuring the particle size distribution and mass-based concentration of polystyrene mixtures containing both nano and microparticles, with the educational aim to underline applicability and limitations of each technique. FINDINGS: Light scattering-based measurements do not have the resolution to distinguish multiple populations in polydisperse samples. Nanoparticle tracking analysis (NTA), nano-flowcytometry (nFCM) and asymmetric flow field flow fractionation hyphenated with multiangle light scattering (AF4-MALS) cannot measure particles in the micrometre range. Static light scattering (SLS) is not able to accurately detect particles below 200 nm, and similarly to transmission electron microscopy (TEM) and flow cytometry (FCM), is not suitable for accurate mass-based concentration measurements. Alternatives for high-resolution sizing and concentration measurements in the size range between 60 nm and 5 µm are tunable resistive pulse sensing (TRPS) and centrifugal liquid sedimentation (CLS), that can bridge the gap between the nanometre and micrometre range.

Sacral neuromodulation in children with urinary and fecal incontinence: a multicenter, open label, randomized, crossover study.

PURPOSE: The clinical benefit of sacral neuromodulation is unclear due to the paucity of randomized trial data. The purpose of this study was to evaluate sacral neuromodulation for management of urinary and fecal incontinence in a pediatric population. MATERIALS AND METHODS: This multicenter, open label, randomized, crossover study included children older than 5 years. After trial stimulation of the S3 root a neuromodulator (InterStim) was implanted on the S3 foramen. Clinical examinations, voiding and bowel diaries, and urodynamic and manometric evaluations were performed at the beginning (t1) and end (t2) of the first period, and at the beginning (t3) and end (t4) of the second period. RESULTS: A total of 33 patients (24 boys) with a mean +/- SD age of 12.22 +/- 5.09 years were randomized. Etiologies were mainly of neurological origin. Incontinence was mixed urinary and fecal in 19 cases, urinary only in 9 and fecal only in 5. Cystometric bladder capacity increased during sacral neuromodulation (delta +24.27 ml vs -37.45 ml, p = 0.01). There was no significant change in other urodynamic or manometric parameters. Overall positive response rate was more than 75% for urinary (81%) and bowel (78%) function. Crossover analysis indicated that sacral neuromodulation is more effective than conservative treatment for both types of incontinence (p = 0.001). CONCLUSIONS: In a pediatric population sacral neuromodulation is effective for bladder and bowel dysfunction and should be considered before irreversible surgery.

A high-intensity highly coherent soft X-ray femtosecond laser seeded by a high harmonic beam.

Synchrotrons have for decades provided invaluable sources of soft X-rays, the application of which has led to significant progress in many areas of science and technology. But future applications of soft X-rays--in structural biology, for example--anticipate the need for pulses with much shorter duration (femtoseconds) and much higher energy (millijoules) than those delivered by synchrotrons. Soft X-ray free-electron lasers should fulfil these requirements but will be limited in number; the pressure on beamtime is therefore likely to be considerable. Laser-driven soft X-ray sources offer a comparatively inexpensive and widely available alternative, but have encountered practical bottlenecks in the quest for high intensities. Here we establish and characterize a soft X-ray laser chain that shows how these bottlenecks can in principle be overcome. By combining the high optical quality available from high-harmonic laser sources (as a seed beam) with a highly energetic soft X-ray laser plasma amplifier, we produce a tabletop soft X-ray femtosecond laser operating at 10 Hz and exhibiting full saturation, high energy, high coherence and full polarization. This technique should be readily applicable on all existing laser-driven soft X-ray facilities.

[Vesico-ureteric reflux treatment by implant of polydimethylsiloxane (Macroplastique): Review of the literature]. / Traitement endoscopique du reflux vésico-urétéral par implant de polydiméthylsiloxane (Macroplastique) : revue de la littérature.

PURPOSE: During the last 25years, endoscopic correction of vesicoureteral reflux (VUR) has become a well-recognized procedure. However, the nature of the bulking agent used has led to many controversies. The objective of this work was to perform a literature review on the use of polydimethylsiloxane (PDMS or Macroplastique) in VUR. METHOD: A PubMed review of the literature since 1996 resulted in the selection of 24 studies of sufficient level of evidence to assess the efficacy and tolerance of Macroplastique in the VUR in adults and children. RESULTS: The overall success rate at 1 year, 2 years and 9-years follow-up was respectively 86-93 %, 80-92 %, and 77-100 %, which confirms the maintenance of good results over time, notably in VUR grade III and above. The success rate was similar for primary and secondary VUR except for total duplicity. Predictive criterias of success were the surgeon's experience, the low grade of VUR, and the absence of previous injection. In comparison with other bulking agents, the higher viscosity and absence of shrinkage of the product increase its reliability. After more than 12years of use, no serious complication has been reported in the literature, reflecting the good tolerance of Macroplastique on the long term. CONCLUSION: The studies published on the use of Macroplastique in VUR confirmed its efficacy, around 85 % of success for all grades, in children and adults as well. The interest of PDMS is linked to its higher viscosity promoting a better reliability and reproducibility of the technique and its non-resorbable nature providing a permanent result, especially valuable in high-grade VUR with anatomical anomaly of the vesicoureteral junction or in VUR secondary to permanent lower urinary tract dysfunction.

[The non-neurogenic neurogenic bladder (Hinman's syndrome) in children: What are pronostic criteria based on a 31 cases multicentric study]. / Critères pronostics du syndrome de Hinman (vessie neurogène non neurogène). Etude multicentrique de 31 cas.

PURPOSE: To identify in children the pejorative estimated criteria of a extreme dysfunctional voiding can leading to a Hinman syndrome. PATIENTS AND METHODS: Retrospective and multicentric study of 31 patients (19 boys and 12 girls), resulting from three universitary medical center (Besancon, Lyon, Nantes), which were divided into three groups according to their evolution: A - forms with a serious uronephrologic outcome. B - intermediate forms with persistent voiding dysfunction. C - forms with uneventful outcome. RESULTS: The initial urologic examination was done, on average, on 6.4 years old children (1-16) and the diagnosis at 12.2 years (1.8-26.9). Eighty-four percent of the patients presented initial urologic symptoms, including 35 % of enuresis and 48 % with large amount of stool in the rectal vault (constipation and encopresis in 93 %). Thirty-five percent undergoing significant familial life stresses. In group A, urinary incontinence was present in 29 %. Sixty-two percent were improved by an intermittent catheterisme and 50 % by anticholinergic drugs solely. In group B, 46 % presented major constipation or encopresis and 60 % an urinary infection. A voiding retraining and an intermittent catheterisme improved 71 and 75 % respectively. In group C, no patient presented initial low urinary infection, encopresis or stool impaction. The boys presented more febrile urinary infection (20 % vs. 8 %), more rectal disorders (encopresis: 31 % vs. 8 %) and less urinary incontinence. In this group voiding retraining improved 67 %. CONCLUSION: The Hinman's syndrome is a serious and unusual outcome of the "dysfunctional elimination syndrome". At diagnosis, the patterns of a pejorative outcome are the male sex, the severe fecal retention, the serious psychological disorders, the delay of diagnosis and treatment.

[Isolated primary nocturnal enuresis: international evidence based management. Consensus recommendations by French expert group]. / Enurésie nocturne primaire isolée: diagnostic et prise en charge. Recommandations par consensus formalisé d'experts.

OBJECTIVE: The causes and treatment of isolated primary nocturnal enuresis (PNE) are the subject of ongoing controversy. We are proposing consensus practical recommendations, based on a formalised analysis of the literature and validated by a large panel of experts. METHODOLOGY: A task force of six experts based its work on the guide for literature analysis and recommendations and recommendation grading of the French Haute Autorité de Santé (formalized consensus process methodological guidelines) to evaluate the level of scientific proof (grade of 1 to 4) and the strength of the recommendations (grade A, B, C) of the publications on PNE. As a result of this, 223 articles from 2003 on were identified, of which only 127 (57 %) have an evaluable level of proof. This evaluation was then reviewed by a 19-member rating group. Several recommendations, poorly defined by the literature, had to be proposed by a professional agreement resulting from a consultation between the members of the task force and those of the rating group. For its final validation, the document was submitted to a reading group of 21 members working in a wide range of specialist areas and practices but all involved in PNE. RESULTS: The definition of PNE is very specific: intermittent incontinence during sleep, from the age of 5, with no continuous period of continence longer than 6 months, with no other associated symptom, particularly during the day. Its diagnosis is clinical by the exclusion of all other urinary pathologies. Two factors must be identified during the consultation: nocturnal polyuria promoted by excessive fluid intake, inverse secretion of vasopressin, snoring and sleep apnoea. It is sensitive to desmopressin; small bladder capacity evaluated according to a voiding diary and the ICCS formula. It may be associated with diurnal hyperactivity of the detrusor (30 %). It is resistant to desmopressin. Problems associated with PNE are: abnormal arousal threshold, attention deficit hyperactivity disorder (ADHD) (10 %), low self-esteem. The psychological component is not very significant. CONCLUSION: PNE is not psychological in origin. The management of this condition includes: evaluating the intrafamilial tolerance and the child's motivation, evaluating the rate, the volume of urine and wet nights using a diurnal and nocturnal diary; education (sufficient fluid intake at the start of the day, decrease in hyperosmolar intake in the evening, regular and complete urination); specific treatments: desmopressin for polyuric forms (expected success rate of 60-70 %), alarms for forms involving small bladder capacity (expected success rate of 60-80 %); alternative treatments and/or treatments combined with the preceding ones, for refractory forms: oxybutinin, tricyclic antidepressants (risk). Results obtained with hypnosis, psychotherapy, acupuncture, homoeopathy or chiropractic are not currently validated (insufficient level of proof).

[Place of the ureteroscopy in the treatment of the children urolithiasis]. / Place de l'urétéroscopie dans le traitement de la lithiase chez l'enfant.

OBJECTIVE: To determine the efficiency and the role of ureteroscopy in the treatment of urolithiasis in children. METHODS: A multicenter retrospective study was conducted between January 2006 and March 2008 in the department of pediatric surgery and urology of Besançon, Lyon and Grenoble. The clinical data of 17 children having benefited from one or more ureteroscopy procedures for urolithiasis were analyzed. These data concerned age, sex, antecedents of metabolic diseases, existence of a malformed uropathy, how the urolithiasis was discovered, therapeutic indications, endoscopic procedures, type of endoscopic treatment for urolithiasis, results and complications. RESULTS: Twenty-six ureteroscopies were carried out for 17 children (eight girls and nine boys) aged between 9 months and 12 years (mean: 5 (1/2) years old) as a first-line treatment or after extracorporeal shock wave lithotripsy. A 6/7,5 Fr semi-rigid ureteroscope was used in all case. Only once, the operator chose a flexible ureteroscope. Lithotripsy was carried out 15 times with YAG Holmium laser, four times with Swiss Lithoclast and six times by simple extraction with a Dormia type basket .A stent probe was left in place after endoscopy for 14 children. The "stone free" rate was 88% with an average hindsight of 11 months. Two incidents without major complication were recorded: a section of the guide by the laser beam and a perforation with extravasation of the contrast agent. CONCLUSION: Ureteroscopy is the first-line treatment in isolated ureteral urolithiasis. On the other hand, LEC remains the treatment of choice for Starghon calculi, since LIC should be offered only in the event of failure of LEC.

Inhibition of proliferation of primary avian fibroblasts through expression of histone H5 depends on the degree of phosphorylation of the protein.

To obtain stable and constitutive expression of histone H5 at levels comparable to those observed in normal chicken erythrocytes, an avian self-inactivating retroviral vector was used to transfer the H5 gene into cells which do not express this protein. The vector, pDAH5, was obtained by removing the CAAT and TATA boxes of the 3'LTR of the avian leukosis virus RAV-2 and inserting the H5 sequence. Infection of QT6 quail cells with the recombinant virus (DAH5) led to the stable integration of the foreign H5 gene at low copy number, to the formation of correctly initiated mRNA transcripts and to the production of H5 protein. The amount of H5 expressed was equivalent to that of a mature chicken erythrocyte. Expression of histone H5 in DAH5 transformed cells, such as QT6 or AEV-ES4, transformed chicken embryo fibroblasts had only slight effects on the growth rate and did not inhibit cell replication. Conversely, the effect of H5 expression on normal quail and chicken fibroblasts was dramatic: cells acquired the aspect of quiescent fibroblasts, grew very slowly, and nuclei looked compacted, often extruded from the cell. The H5 histone produced in QT6-transformed cells was found to be phosphorylated while in normal chicken fibroblasts the protein lacked this posttranslational modification. It is proposed that the chromatin-condensing role of histone H5 is inhibited by its phosphorylation.

Effect of acute hypoxia on central fatigue during repeated isometric leg contractions.

To determine whether hypoxia has a direct influence on the central command independently of the working muscles, 16 subjects performed intermittent isometric unilateral knee extensions until exhaustion either in normobaric hypoxia (inspired O(2) fraction=0.11, arterial oxygen saturation approximately 84%) or in normoxia while the knee extensor muscles were exposed to circulatory occlusion with a 250 mmHg cuff. Among the subjects, 11 also performed the tests in hypoxia and normoxia without occlusion. Single electrical stimulations were regularly delivered to the femoral nerve to measure the changes in the knee extensor peak twitch force. With the cuff, the average slope of decrease in peak twitch did not depend on the inspired oxygen fraction. Performance was slightly but significantly lower during hypoxia than in normoxia (8.2+/-2.6 vs 9.4+/-3.1 repetitions, P<0.05) with the cuff on. The number of repetitions was much higher during hypoxia with maintaining leg blood flow (15.6+/-4.5 repetitions) than with circulatory occlusion in normoxia. In conclusion, this study showed that a direct effect of hypoxia in reducing the motor drive to the working muscles exists but this effect is moderate.

Landscape, herd management and within-herd seroprevalence of Toxoplasma gondii in beef cattle herds from Champagne-Ardenne, France.

Toxoplasmosis is the most common parasitic zoonosis worldwide. Its prevalence and impact on human health are highly variable geographically. Humans may be infected by ingesting oocysts from the environment, or bradyzoits contained in meat products from various domestic species, thus data on the dynamics of toxoplasmosis in domestic herds is needed. However, few information is available on the factors that determine the level of infection of cattle herds. In this study, we aimed to estimate within-herd seroprevalence of toxoplasmosis in bovine herds and analyze its variability in relation with environmental characteristics and herd management. We tested the presence of anti Toxoplasma gondii antibodies in 1329 cattle from 24 beef herds in the Champagne-Ardenne region, using the Modified Agglutination Test. Information on herds was collected using a questionnaire. After describing the relationships between explanatory variables, we built a multivariate model using logistic Generalized Linear Models (GLMs) and estimated parameters with a Generalized Estimating Equations (GEE) procedure. The crude seroprevalence at threshold 1:24 equalled 7.8%, which is low, however positive individuals were found in 21 out of 24 herds. The final multivariate model showed that within-herd seroprevalence was highest in herds that were both small and isolated. The presence of cats modified the age-prevalence relationship: maximal seroprevalence was observed in oldest cows in farms without cats, and in youngest individuals in farms with cats. Finally, using a natural water point on pastures was associated to a high within-herd seroprevalence (Odds-Ratio: 1.93). Cows are often exposed to toxoplasmosis, however landscape characteristics (water point, isolation) and herd management (herd size, cats) may affect seroprevalence. Our results may help to find ways of reducing T. gondii prevalence in cattle.

Detection of Toxoplasma gondii oocysts in water: proposition of a strategy and evaluation in Champagne-Ardenne Region, France.

Water is a vehicle for disseminating human and veterinary toxoplasmosis due to oocyst contamination. Several outbreaks of toxoplasmosis throughout the world have been related to contaminated drinking water. We have developed a method for the detection of Toxoplasma gondii oocysts in water and we propose a strategy for the detection of multiple waterborne parasites, including Cryptosporidium spp. and Giardia. Water samples were filtered to recover Toxoplasma oocysts and, after the detection of Cryptosporidium oocysts and Giardia cysts by immunofluorescence, as recommended by French norm procedure NF T 90-455, the samples were purified on a sucrose density gradient. Detection of Toxoplasma was based on PCR amplification and mouse inoculation to determine the presence and infectivity of recovered oocysts. After experimental seeding assays, we determined that the PCR assay was more sensitive than the bioassay. This strategy was then applied to 482 environmental water samples collected since 2001. We detected Toxoplasma DNA in 37 environmental samples (7.7%), including public drinking water; however, none of them were positive by bioassay. This strategy efficiently detects Toxoplasma oocysts in water and may be suitable as a public health sentinel method. Alternative methods can be used in conjunction with this one to determine the infectivity of parasites that were detected by molecular methods.