COVID-19 wastewater based epidemiology: long-term monitoring of 10 WWTP in France reveals the importance of the sampling context.

SARS-CoV-2 wastewater-based epidemiology (WBE) has been advanced as a relevant indicator of distribution of COVID-19 in communities, supporting classical testing and tracing epidemiological approaches. An extensive sampling campaign, including ten municipal wastewater treatment plants, has been conducted in different cities of France over a 20-week period, encompassing the second peak of COVID-19 outbreak in France. A well-recognised ultrafiltration - RNA extraction - RT-qPCR protocol was used and qualified, showing 5.5 +/- 0.5% recovery yield on heat-inactivated SARS-CoV-2. Importantly the whole, solid and liquid, fraction of wastewater was used for virus concentration in this study. Campaign results showed medium- to strong- correlation between SARS-CoV-2 WBE data and COVID-19 prevalence. To go further, statistical relationships between WWTP inlet flow rate and rainfall were studied and taken into account for each WWTP in order to calculate contextualized SARS-CoV-2 loads. This metric presented improved correlation strengths with COVID-19 prevalence for WWTP particularly submitted and sensitive to rain. Such findings highlighted that SARS-CoV-2 WBE data ultimately require to be contextualized for relevant interpretation.

Comparison of two filtration-elution procedures to improve the standard methods ISO 10705-1 & 2 for bacteriophage detection in groundwater, surface water and finished water samples.

AIM: To select a reliable method for bacteriophage concentration prior detection by culture from surface water, groundwater and drinking water to enhance the sensitivity of the standard methods ISO 10705-1 & 2. METHODS AND RESULTS: Artificially contaminated (groundwater and drinking water) and naturally contaminated (surface water) 1-litre samples were processed for bacteriophages detection. The spiked samples were inoculated with about 150 PFU of F-specific RNA bacteriophages and somatic coliphages using wastewater. Bacteriophage detection in the water samples was achieved using the standard method without and with a concentration step (electropositive Anodisc membrane or a pretreated electronegative Micro Filtration membrane, MF). For artificially contaminated matrices (drinking and ground waters), recovery rates using the concentration step were superior to 70% whilst analyses without concentration step mainly led to false negative results. Besides, the MF membrane presented higher performances compared with the Anodisc membrane. CONCLUSION: The concentration of a large volume of water (up to one litre) on a filter membrane avoids false negative results obtained by direct analysis as it allows detecting low number of bacteriophages in water samples. SIGNIFICANCE AND IMPACT OF THE STUDY: The addition of concentration step before applying the standard method could be useful to enhance the reliability of bacteriophages monitoring in water samples as bio-indicators to highlight faecal pollution.

Incidence and prevalence of juvenile idiopathic arthritis in Catalonia (Spain).

OBJECTIVE: To ascertain the incidence and prevalence of juvenile idiopathic arthritis (JIA) in Catalonia (autonomous region in northeast Spain), examined according to the currently established disease subtypes. METHODS: Before initiating the study, we conducted an educational programme on paediatric rheumatology, addressed to all general paediatricians in Catalonia. A 2-year (2004-2006), prospective, population-based study was then carried out to determine the incidence of JIA. Prospective and retrospective data retrieval was performed to calculate prevalence. The International League of Associations for Rheumatology (ILAR, Edmonton revision) classification criteria were applied. RESULTS: Over the study period, 145 new cases of JIA were diagnosed. The mean annual incidence was 6.9/105 children aged less than 16 years (range 5.8-8.1 years; 9.0 years for girls and 4.8 years for boys). On separate analysis of patients ≤ 6 and > 6 years, the distribution in younger children was found to be similar for both girls and boys, whereas in older children, most girls belonged to the oligoarthritis and polyarthritis subgroups, and boys to the enthesitis-related arthritis and undifferentiated subgroups. The calculated prevalence of JIA (31 October 2006) was 39.7 (36.1-43.7)/105 children younger than 16. The relative risk of girls having JIA was 2.1 [95% confidence interval (CI) 1.7-2.7, p < 0.001]. In 70% of patients, the diagnosis was established before the age of 7. Subgroup distribution of prevalent cases mirrored that of incident cases. CONCLUSION: This is the first population-based study on the epidemiology of JIA in Catalonia. Incidence and prevalence rates are lower than those reported for several areas in Nordic countries of Europe. Oligoarthritis was the most common subtype.

Adenovirus, MS2 and PhiX174 interactions with drinking water biofilms developed on PVC, cement and cast iron.

Biofilms colonizing pipe surfaces of drinking water distribution systems could provide habitat and shelter for pathogenic viruses present in the water phase. This study aims (i) to develop a method to detect viral particles present in a drinking water biofilm and (ii) to study viral interactions with drinking water biofilms. A pilot scale system was used to develop drinking water biofilms on 3 materials (7 cm(2) discs): PVC, cast iron and cement. Biofilms were inoculated with viral model including MS2, PhiX174 or adenovirus. Five techniques were tested to recover virus from biofilms. The most efficient uses beef extract and glycine at pH = 9. After sonication and centrifugation, the pH of the supernatant is neutralized prior to viral analysis. The calculated recovery rates varied from 29.3 to 74.6% depending on the virus (MS2 or PhiX174) and the material. Applying this protocol, the interactions of virus models (MS2 and adenovirus) with drinking water biofilms were compared. Our results show that adsorption of viruses to biofilms depends on their isoelectric points, the disc material and the hydrodynamic conditions. Applying hydrodynamic conditions similar to those existing in drinking water networks resulted in a viral adsorption corresponding to less than 1% of the initial viral load.

Multiple sclerosis association study with the TENR-IL2-IL21 region in a Spanish population.

Polymorphisms from the TENR-IL2-IL21 block in the 4q27 chromosome were recently associated with type 1 diabetes, celiac disease, rheumatoid arthritis and psoriasis. We undertook this study to investigate the potential role of polymorphisms rs3136534, rs6822844 and rs2069762 (-330 T/G IL2) in multiple sclerosis (MS) (805 patients of Spanish Caucasian origin and 952 health controls). We did not find evidence for association with any single nucleotide polymorphisms (SNPs) tested. Allele and genotype frequencies of the SNPs, which were studied, were similar in DRB1*15-positive or DRB1*15-negative patients. After stratification of MS patients by clinical course, a weak association was observed with rs2069762 G allele and haplotype bearing this allele with secondary progressive MS, although these cases represent 22% of the MS cases. Our results did not show major influence of TENR-IL2-IL21 locus on susceptibility or disease progression in MS. However, we could not exclude completely the effect in MS for this region. Additional studies, using much larger sample sizes and analysis of additional polymorphisms in the gene and its flanking region, will be required to ascertain their contributions to MS susceptibility.

Assessing the cost of groundwater pollution: the case of diffuse agricultural pollution in the Upper Rhine valley aquifer.

This paper presents an assessment of the costs of diffuse groundwater pollution by nitrates and pesticides for the industrial and the drinking water sectors in the Upper Rhine valley, France. Pollution costs which occurred between 1988 and 2002 are described and assessed using the avoidance cost method. Geo-statistical methods (kriging) are then used to construct three scenarios of nitrate concentration evolution. The economic consequences of each scenario are then assessed. The estimates obtained are compared with the results of a contingent valuation study carried out in the same study area ten years earlier.

Detection of mediastinitis after heart transplantation by gallium-67 scintigraphy.

We report the findings of a patient with post-cardiac transplant mediastinitis detected by 67Ga-citrate imaging. Fever and leukocytosis were the first clinical signs suggesting infection. The usual diagnostic modalities, including CT and ultrasound, failed to identify the site of infection. A 67Ga scan showed intense abnormal uptake behind the sternum. The site of uptake was shown by necropsy to be necrotic tissue involving cardiac sutures, pulmonary arteries, and the aorta due to infection with Haemophilus aphrophilus.

Comparison of seven RNA extraction methods on stool and shellfish samples prior to hepatitis A virus amplification.

When choosing an extraction method, two parameters have to be considered: recovery of the viral material and elimination or inactivation of inhibitory substances. Seven techniques for extracting hepatitis A virus (HAV) from stool and shellfish samples were compared, in order to identify the protocol most suited to both types of sample and with the best extraction yield. The protocols tested were either techniques for the recovery and purification of total RNA, such as RNAzol, PEG-CETAB, GTC-silica and Chelex, or techniques for isolating specifically HAV using a nucleotide probe or a monoclonal antibody. For stool samples, RNAzol, PEG-CETAB, and magnetic beads with antibody allowed detection of the virus in 11/12 and 12/12 of samples. For shellfish samples, three protocols allowed RNA to be extracted in 90% of cases, RNAzol, PEG-CETAB, and GTC-silica. Their rapidity and low cost make RNAzol and GTC-silica the most suitable for routine diagnostic testing. reserved.

Case report: organic brain syndrome due to multiple tumour emboli.

A case of rapid neurological deterioration in a 60-year-old man with oesophageal carcinoma is reported. He suffered two episodes of global cerebral dysfunction following traumatic endoscopic manipulation. Autopsy findings showed multiple tumour emboli which occluded medium and small-sized vessels in cortical and subcortical areas, resulting in multiple microinfarcts.

Systemic onset juvenile chronic arthritis, polyarticular pattern and hip involvement as markers for a bad prognosis.

OBJECTIVE: To explore all the common clinical and biological variables that are characteristic of Systemic onset Juvenile Chronic Arthritis (SoJCA) in order to determine which of them are suitable as predictors of a bad articular outcome (persistence of inflammatory symptoms and/or established limitation of the range of motion (ROM). MATERIAL AND METHODS: Clinical charts for 124 SoJCA patients were retrospectively reviewed. From them, 91 were finally included in the study because they had all of the clinical and biological data at disease onset properly recorded. All have been followed for at least 3 years since the beginning of the disease. Data collected at onset, and after 3 and 6 months of the disease included: 1) systemic symptoms; 2) joint involvement, using both the usual articular count and the value of an articular index (Helsinki Index = HI) which intentionally excludes those joints that are not uniformly recorded in clinical charts; and 3) biological data. HI was used to separate the patients into two groups. When applied 3 years after the disease onset, HI > or = 10 represented a bad articular outcome while HI < 10 meant a good prognosis. SPSS for Windows 6.1 was used for both the univariate and multivariate analyses. RESULTS: From the multivariate logistic regression analysis, two different "clusters" of clinical data were found to be the best predictors of a bad articular outcome. A bad prognosis was linked at onset with the presence of generalized lymphadenopathies, age < 8 years and an HI > 6; at six months a bad outcome was linked with the presence of a polyarticular pattern plus hip involvement. CONCLUSION: Clinical parameters at the beginning of the disease were shown to be extremely useful in predicting the articular outcome of SoJCA. Therefore, they could constitute a good instrument to help clinicians tailor the best therapy for their patients.

Concomitant rheumatoid arthritis and amyotrophic lateral sclerosis. A puzzle illustrated by a new case.

We report a case of amyotrophic lateral sclerosis in a patient with rheumatoid arthritis. Only three similar cases have been reported. Our case illustrates the diagnostic difficulties raised by early amyotrophic lateral sclerosis responsible for localized or unusual manifestations. Occurrence of the two diseases in the same patient is probably due to chance alone.

Oxidant-antioxidant imbalance in blood of children with juvenile rheumatoid arthritis.

Juvenile rheumatoid arthritis (JRA) is the most commonly diagnosed rheumatic disease in children and may represent not a single disease, but rather a syndrome of diverse aetiologies in which inflammation is an exceedingly complex process. Oxidative free radical production at inflammation sites contributes to tissue damage and could also play a significant role in the pathogenesis of JRA. The aim of this study was to evaluate the antioxidant status and lipid peroxidation parameters related to the clinical form of JRA. Plasma malondialdehyde and hydroperoxide concentrations in children with polyarticular and systemic JRA subtypes were significantly higher than in controls. Plasma vitamin E and beta-carotene levels of the JRA children were lower in the three forms compared with healthy children. Patients with JRA present an imbalance in the oxidant-antioxidant system that manifests clearly in the polyarticular and systemic forms through an increase in lipoperoxidation products and significant decrease in the lipid-soluble antioxidants vitamin E and beta-carotene.

Immunocytochemical detection of dendritic cell by S-100 protein in the chicken.

Positivity for S-100 protein in paraffin embedded chicken lymphoid tissue was found by using a polyclonal antibody against whole bovine S-100 protein. The S-100 protein-containing cells were observed in the locations which have been reported to contain avian dendritic cells such as the medulla of the bursal follicles, and the germinal centers and T-dependent areas in the spleen, Peyer's patches, caecal tonsil and Harderian gland. Positive cells were also found in the location where ellipsoid associated cell have been described, and between epithelial cells covering the Peyer's patches and the caecal tonsil, as well as between the cells lining the ducts within the Harderian gland. Macrophages were devoid of immunostaining. Our results confirm the location described elsewhere for chicken dendritic cells and indicate that S-100 protein can be considered as a cell marker for the identification of the chicken dendritic cell. Intraepithelial positive cells may be interdigitating dendritic cells in an unusual location (their function being the transport of the antigen from the epithelium to the diffuse lymphoid tissue), or cytotoxic T-lymphocytes which, in mammals, are immunoreactive for S-100 protein.

Local immune response in the chicken Harderian gland to antigen given by different ocular routes.

The effectiveness of three ocular routes of antigen administration to produce a local immune response in the Harderian gland was studied. The routes were by eyedrop, injection into the ocular conjunctiva and injection into the nictitating membrane. The antigen was observed in the cytoplasm of macrophages located within the lymphoid tissue only after the injection into the nictitating membrane. The numbers of germinal centres and plaque forming cells found in the gland after injection into the nictitating membrane was higher than the numbers observed following the other two ocular applications. These findings indicate that the injection of the antigen into the nictitating membrane is the most effective ocular route for producing a local immune response in the Harderian gland.

[Chest radiology in rheumatoid arthritis. A study of 89 patients (author's transl)]. / La radiología de tórax en la artritis reumatoidea. Estudio de 89 enfermos.

Chronic interstitial pneumonia, necrobiotic nodules and pleural lesions are felt to be extraarticular manifestations of rheumatoid arthritis (RA). In a prospective study, posteroanterior and lateral chest X-ray films were taken in 89 patients with classical or definite RA (15 male and 74 female), of whom 64 (71.9%) were seropositive. The findings were compared with those of 103 age and sex matched controls. The chest X-rays were blindly evaluated by two of the authors (a pneumologist and a radiologist), and the presence of an interstitial pattern, nodules or pleural lesions was recorded. The following results were obtained: Interstitial pattern, in patients with RA, 5/89 (5.6%), and in controls, 5/103 (4.6%). Pulmonary nodules, in RA, 2/89 (2.2%), and in controls, 2/103 (1.9%). Pleural adhesions, in RA, 32/89 (35.9%), and in controls, 24/103 (23.5%). The results of the present series demonstrate that radiological pulmonary lesions are not more frequent in RA than in controls, similar conclusions having been reached by other authors.

[Circulating immune complexes-like material and complement in rheumatoid arthritis and ankylosing spondylitis (author's transl)]. / Inmunocomplejos circulantes y complemento en la artritis reumatoidea y en la espondilartritis anquilosante.

Circulating immune complexes (IC) in 52 rheumatoid arthritis (RA) seropositive, 25 RA seronegative and 32 ankylosing spondylitis (AS) patients were assayed by two methods: 1) precipitation with a final 2% polyethylene glycol (PEG) concentration with immunoglobulins (Igs) quantitation in the precipitate and 2) the ability of IC to consume complement, evaluating conversion products from human C3 by bidimensional rocket immunoelectrophoresis. CH50, C3, C4, C3PA and circulating C3 conversion products were quantitated. IC precipitates with 2% PEG contained three classes of Igs: IgG, IgM and IgA, the latter being the most rare. In the control group Igs level precipitated with 2% PEG were: 2% PEG-IgG 2.5 mg/100 ml +/- 2; 2% PEG-IgM 0.72 mg/100 ml +/- 1.09; 2% PEG-IgA 0.04 mg/100 ml +/- 0.18. With 2% PEG IC-like material was detected in 58.5% of RA seropositive patients but none generated C3 conversion products. With neither methods was IC-like material detected in the RA seronegative group and none with 2% PEG in AS, either. However, in the latter, 21.8% generated C3 conversion products and 18.7% circulating C3 conversión products. In the RA seropositive patients no correlation was observed between the levels of 2% PEG-IgG and 2% PEG-IgM and the titre of antinuclear antibodies (ANA) nor between 2% PEG-IgG and that of IgG in serum. However, there was a low correlation between 2% PEG-IgM and that of IgM in serum and between 2% PEG-IgG and 2% PEG-IgM with rheumatoid factor (RF). It should be stressed that the value of the PEG method in the detection of IC-like material must be reconsidered, since using a 2% final concentration and a specific evaluation of Igs and complement components in the precipitate, it may be a method easy to perform and useful for clinical purposes.

[Focal myositis in children. Apropos of a case]. / Myosite focale chez l'enfant. A propos d'un cas.

An eleven year old boy presented with localized swelling of the left calf which had been developing for more than 5 years. Another smaller mass was found in the pretibial area. There was an invincible equinus deformity of the foot. Surgical biopsies revealed characteristic histological features of focal myositis showing necrotic and regenerating muscle fibers, randomly distributed, with inflammatory infiltration. An intense fibrotic reaction was observed. This constitutes to our knowledge the first reported case of focal myositis in which development of muscle sclerosis was so important that it was necessary to perform tenotomies one year after excision of the principal tumor in order to restore normal gait. A nearly two-years follow-up does not give evidence of recurrent lesion.

[Subdural empyema due to Mycoplasma hominis following epidural anesthesia]. / Empiema subdural por Mycoplasma hominis tras anestesia epidural.

INTRODUCTION: In the literature there are sporadic reports of spinal epidural abscesses after epidural anaesthesia (Staphylococcus aureus in 82%), whilst subdural empyemas are more often related to ear and sinus conditions. CLINICAL CASE: A 32 year old woman with a clinical history of migraine and symmetrical frontal atrophy on a previous cerebral CT scan, after Caesarean section under epidural anaesthesia, presented with orthostatic headache two days later. On the fourth day it had become constant and she had a high temperature which was considered to be caused by infection of the surgical wound. Neurological examination was found to be normal, the CT scan was inconclusive and the CSF showed a lymphocytic pleocytosis without consumption of glucose. In view of her worsening clinical condition on the ninth day, in the absence of a cutaneous focus and on suspicion of a para-meningeal infective focus, lumbar MR was done and found to be normal, and cerebral MR which showed images compatible with a right fronto-parietal subdural empyema. After a parietal craniotomy and culture of the surgical specimen, colonies of Mycoplasma hominis were grown, similar to those grown from the exudates of the abdomical surgical wound. Treatment was started with ciprofloxacine. CONCLUSION: We consider that following epidural anaesthesia the patient developed hypotension of the CSF with a secondary subdural hematoma or hygroma and this became infected by hematogenous spread of Mycoplasma hominis.

[Four new cases and a review of the literature concerning the migraine with csf pleocytosis syndrome]. / Síndrome de migraña con pleocitosis de LCR: aportación de 4 nuevos casos y revisión de la literatura.

The syndrome known as migraine with cerebrospinal fluid (CSF) pleocytosis is a clinical syndrome of controversial pathogenesis characterized by typical migraine headaches occurring in a fixed period of time along with lymphocyte pleocytosis in CSF which persists during intercritical periods. We present four new cases of this syndrome diagnosed in our service and we review those cases published in the literature in which there were no personal headache antecedents, attempting in this way to separate this syndrome from other types of migraine as included in the present classification. There also exists the possibility that it might be a question of a secondary process, the pathogenesis of which may play a part in the activation of the humoral immune system, given the frequent existence of immunogenic antecedents the previous days as well as that of a humoral reaction expressed as an increase in IgG levels in CSF with normal blood electrophoresis.