Synovial fluid but not plasma interleukin-8 is associated with clinical severity and inflammatory markers in knee osteoarthritis women with joint effusion.

Several cytokines and adipokines are related to clinical severity and progression in knee osteoarthritis. The aim of this study was to evaluate the associations of IL-8 with clinical severity and with local and systemic adipokines and cytokines. This is a Cross-sectional study including 115 women with symptomatic primary knee osteoarthritis with ultrasound-confirmed joint effusion. Age, symptoms duration and body mass index were collected. Radiographic severity was evaluated according to Kellgren-Lawrence. Pain and disability were assessed by Lequesne and Knee injury and Osteoarthritis Outcome Score pain, symptoms and function scales. Three inflammatory markers and five adipokines were measured by ELISA in serum and synovial fluid. Partial correlation coefficient (PCC) and corresponding 95% confidence interval were used to evaluate association. Synovial fluid IL-8 was significantly associated with clinical severity scales. After controlling for potential confounders, associations measured by a Partial Correlation Coefficient (PCC) remained essentially unaltered for Lequesne (PCC = 0.237), KOOS pain (PCC = - 0.201) and KOOS symptoms (PCC = - 0.209), KOOS function (PCC = - 0.185), although the later did not reach statistical significance. Also in synovial fluid samples, associations were found between IL-8 and TNF (PCC = 0.334), IL6 (PCC = 0.461), osteopontin (PCC = 0.575), visfatin (PCC = 0.194) and resistin (PCC = 0.182), although significance was not achieved for the later after statistical control for confounders. None of these associations were detected in serum. In conclusion, IL-8 was associated with clinical severity, inflammatory markers and adipokines in synovial fluid, but not in blood. Although the reported associations are weak to moderate in magnitude, these findings reinforce the notion that local and not systemic inflammation is more relevant to clinical severity in knee OA women with joint effusion.

Comparison of methods for concentrating human adenoviruses, polyomavirus JC and noroviruses in source waters and drinking water using quantitative PCR.

Human adenovirus and JC polyomavirus have been proposed as viral indicators of human faecal contamination of water. This study compared concentration and nucleic acid extraction methods and defines a protocol for quantifying human adenoviruses (HAdV), JC polyomavirus (JCPyV) and noroviruses (NoV) in source and drinking water. River water samples and spiked tap water samples were used to evaluate virus recovery, applying quantitative PCR (qPCR) to five concentration methods. In the case of 10-L samples, the use of ultrafiltration cartridges produced acceptable recoveries for HAdV and JCPyV, but they were inefficient for noroviruses and could not be applied to high-volume and river water samples with medium turbidity. The glass wool method with pre-acidification gave similar recoveries and made it possible to detect NoV. In the case of 50-L samples, the method that produced the highest recovery efficiency and applicability was glass wool filtration. Comparing different sample volumes of a river used as source water showed that the largest number of viruses were quantified when lower volumes (1L) were tested (1.5 x 10(4) HAdV genome copies (GC)/L and 2.8 x 10(3) JCPyV GC/L). The methods developed are easy to standardize and may be valuable tools for the control of viral contamination in source water and for assessing the efficiency of virus removal in drinking water treatment plants.

Analysis of adenoviruses and polyomaviruses quantified by qPCR as indicators of water quality in source and drinking-water treatment plants.

Three drinking-water treatment plants were analyzed for the presence of human adenoviruses (HAdV) and JC polyomavirus (JCPyV), previously suggested as viral contamination indicators, in order to define their water quality in relation to the presence of viral pathogens and the efficiency of the treatments applied. The 90% of the river water samples had positive results of HAdV (10(1)-10(4) genome copies (GC)/L); and 48%, of JCPyV (10(0)-10(3)GC/L). Lower concentrations of HAdV and JCPyV were found in different treatment steps of the plants in absence of bacterial standards. Virus removal efficiencies were higher than 5 logs in plants 1 and 3 and could be quantified as >2 logs in plant 2. However, three post-chlorinated samples from plants 2 and 3 (11%) were found to be positive for HAdV by qPCR, but did not show infectivity in the cell cultures assayed. Simple methods based on the adsorption-elution of viruses from glass wool give low-cost and efficient virus recovery from source water and large-volume water samples. Quantification of JCPyV and HAdV using qPCR is useful for evaluating virus removal efficiency in water treatment plants, identification of Hazard Analysis and Critical Control Points (HACCP) and as a molecular index of the virological quality of water. Though infectivity is not guaranteed when using qPCR techniques in water treated with disinfection processes, the quality of source water, where viruses have proved to have infective capabilities, and the removal efficiency of viral particles may be efficiently quantified.

Distribution of human polyomaviruses, adenoviruses, and hepatitis E virus in the environment and in a drinking-water treatment plant.

Large numbers of viruses are excreted in human feces and urine, which even at low concentrations may cause illness when ingested. Some of these viruses have not been traditionally monitored in terms of waterborne diseases and are considered emergent viruses, such as hepatitis E virus (HEV) and JC and BK polyomavirus (JCPyV and BKPyV). The high prevalence of human adenoviruses (HAdV) and polyomaviruses, which both show DNA genomes, in sewage from widely divergent areas has suggested the relevance of evaluating these viruses as possible indicators of viral contamination. The concentration of these viruses was analyzed in sewage and river water and after treatment in a drinking-water treatment plant including chlorination, flocculation, ozonation, and granulate active carbon (GAC) filtration. Samples of GAC-filtered water were collected before a second chlorination treatment. The river used as a source of fresh water presented an average concentration of 2.6 x 10(1) JCPyV and 4 x 10(2) HAdV GC (genome copies)/L. A removal of 2 logarithms (99%) of HAdV and JCPyV was observed in the drinking-water treatment plant. All the GAC-filtered water samples studied contained HAdV, with a mean value of 4.3 HAdV GC/L. HEV strains belonging to genotype 3 were frequently detected in low concentrations in urban sewage and in biosolids or sewage containing swine feces but not in the river water samples studied. The detection of viruses by molecular techniques is useful for genetically describe emergent viruses in community wastewaters and water supplies. Quantification of JCPyV and HAdV using quantitative real-time PCR (QPCR) may be useful for evaluating virus removal efficiency in water treatment plants and as an index of the virological quality of water and of the potential presence of human viruses.

Quantification and stability of human adenoviruses and polyomavirus JCPyV in wastewater matrices.

Human adenoviruses (HAdV) and human polyomavirus JCPyV have been previously proposed as indicators of fecal viral contamination in the environment. Different wastewater matrices have been analyzed by applying real-time quantitative PCR procedures for the presence, quantity, and stability of a wide diversity of excreted HAdV and JCPyV. High quantities of HAdV and JCPyV were detected in sewage, effluent wastewater, sludge, and biosolid samples. Both viruses showed high stability in urban sewage. These results confirm the suitability of both viruses as indicators of human fecal viral pollution.

Identification of human and animal adenoviruses and polyomaviruses for determination of sources of fecal contamination in the environment.

The Adenoviridae and Polyomaviridae families comprise a wide diversity of viruses which may be excreted for long periods in feces or urine. In this study, a preliminary analysis of the prevalence in the environment and the potential usefulness as source-tracking tools of human and animal adenoviruses and polyomaviruses has been developed. Molecular assays based on PCR specifically targeting human adenoviruses (HAdV), porcine adenoviruses (PAdV), bovine adenoviruses (BAdV), and bovine polyomaviruses (BPyV) were applied to environmental samples including urban sewage, slaughterhouse, and river water samples. PAdV and BPyV were detected in a very high percentage of samples potentially affected by either porcine or bovine fecal contamination, respectively. However, BAdV were detected in only one sample, showing a lower prevalence than BPyV in the wastewater samples analyzed. The 22 slaughterhouse samples with fecal contamination of animal origin showed negative results for the presence of HAdV. The river water samples analyzed were positive for the presence of both human and animal adenoviruses and polyomaviruses, indicating the existence of diverse sources of contamination. The identities of the viruses detected were confirmed by analyses of the amplified sequences. All BPyV isolates showed a 97% similarity in nucleotide sequences. This is the first time that PAdV5, BAdV6, and BPyV have been reported to occur in environmental samples. Human and porcine adenoviruses and human and bovine polyomaviruses are proposed as tools for evaluating the presence of viral contamination and for tracking the origin of fecal/urine contamination in environmental samples.

[Effects on health of water and food contamination by emergent human viruses]. / Efectos sobre la salud de la contaminación de agua y alimentos por virus emergentes humanos.

The development of molecular technologies applied to environmental studies has shown that even in highly industrialized countries there is a high prevalence of viruses in the environment that represents an important impact on public health and substantial economic losses mainly related to the transmission of viruses through water and food. Significant concentrations of viruses are detected in the water flowed to the environment and in the biosolids generated in wastewater treatment plants. This work describes the general characteristics of the environmental contamination by viruses principally by emergent viruses, with a special emphasis on the hepatitis E virus (HEV) and the human polyomaviruses as the environmental contaminants more recently identified in industrialized countries. It has been shown that there is a high prevalence of the human polyomaviruses BKV and JCV in urban sewage in all studied countries, implying a potential transmission of these viruses and their potential oncogenic genes through the oral route. Recent studies have shown that the epidemiological pattern of the HEV infection in industrialized countries is complex and that a diversity of HEV strains simultaneously infects the population. The control of the viral contamination requires the standardization of molecular techniques and the development of a surveillance program for the evaluation of the viral parameters and to reduce the dissemination of already established diseases and emergent viral infections.

Nested multiplex PCR assay for detection of human enteric viruses in shellfish and sewage.

Environmental samples and contaminated shellfish present frequently low concentrations of more than one viral species. For this reason, a nested multiplex RT-PCR was developed for the detection of adenoviruses, enteroviruses and hepatitis A viruses in different environmental samples such as urban sewage and shellfish. This assay will save time and cost for detection of these enteric viruses with a smaller sample volume, which otherwise can be a limiting factor in routine analysis. The limit of detection was approximately 1 copy for adenovirus and 10 copies for enterovirus and hepatitis A virus per PCR reaction using titrated cell-cultured viruses as template material. In shellfish and environmental samples, this multiplex PCR was optimized to detect all three viruses simultaneously when the concentration of each virus was equal or lower than 1000 copies per PCR reaction. This is the level found predominantly in the environment and in shellfish when the numbers of fecal bacterial and phage indicators are low. The detection of human adenoviruses by PCR has been suggested as a molecular index of fecal contamination of human origin in the environment and food and the multiplex assay developed may be a tool for evaluating the presence of viral contamination in shellfish and water and to expand microbiological control to include viral markers.