title: Qualitative and Quantitative Assessment of Hepatitis A Virus in Wastewaters in Tunisia.
abstract: Hepatitis A causes substantial morbidity in both industrialized and non-industrialized countries and represents an important health problem in several southern Mediterranean countries. The objectives of the study were as follows: (a) to assess the occurrence of hepatitis A virus (HAV) in Tunisia through the monitoring of urban wastewaters collected at wastewater treatment plants (WTPs); (b) to characterize environmental strains; and (c) to estimate the viral load in raw and treated sewages, in order to evaluate the potential impact on superficial waters receiving discharges. A total of 150 raw and treated wastewaters were collected from three WTPs and analyzed by both qualitative (RT-PCR/nested) and quantitative (qRT-PCR) methods. Of these, 100 (66%) were found to be positive for HAV by the qualitative assay: 68.3% in influents and 64.7% in effluents. The vast majority of HAV sequences belonged to sub-genotype IA, with 11 different strains detected found to be identical to clinical strains isolated from Tunisian patients with acute hepatitis. Five unique variants were also detected, not previously reported in clinical cases. Only two IB strains were found, confirming the rarity of this sub-genotype in this country. The results of the present study indicate a wide circulation of the pathogen in the population, most probably in the form of asymptomatic infections, a finding consistent with the classification of the country as having intermediate/high endemicity. Quantitative data showed high viral loads in influents (3.5E+05 genome copies/liter, mean value) as well as effluents (2.5E+05 genome copies/liter, mean value), suggesting that contaminated water could be a critical element in transmission.
authors.affiliation: Laboratory of Transmissible Diseases and Biological Active Substances, Faculty of Pharmacy, University of Monastir, Monastir, Tunisia.
authors.forename: A
authors.initials: A
authors.lastname: Béji-Hamza
authors.affiliation: Laboratory of Transmissible Diseases and Biological Active Substances, Faculty of Pharmacy, University of Monastir, Monastir, Tunisia.
authors.forename: H
authors.initials: H
authors.lastname: Khélifi-Gharbi
authors.affiliation: Laboratory of Transmissible Diseases and Biological Active Substances, Faculty of Pharmacy, University of Monastir, Monastir, Tunisia.
authors.forename: M
authors.initials: M
authors.lastname: Hassine-Zaafrane
authors.affiliation: Department of Environment and Primary Prevention, Istituto Superiore di Sanità, Viale Regina Elena 299, 00161, Rome, Italy.
authors.forename: S
authors.initials: S
authors.lastname: Della Libera
authors.affiliation: Department of Environment and Primary Prevention, Istituto Superiore di Sanità, Viale Regina Elena 299, 00161, Rome, Italy.
authors.forename: M
authors.initials: M
authors.lastname: Iaconelli
authors.affiliation: Department of Environment and Primary Prevention, Istituto Superiore di Sanità, Viale Regina Elena 299, 00161, Rome, Italy.
authors.forename: M
authors.initials: M
authors.lastname: Muscillo
authors.affiliation: Department of Environment and Primary Prevention, Istituto Superiore di Sanità, Viale Regina Elena 299, 00161, Rome, Italy.
authors.forename: S
authors.initials: S
authors.lastname: Petricca
authors.affiliation: Department of Infectious, Parasitic and Immune-Mediated Diseases, Istituto Superiore di Sanità, Viale Regina Elena 299, 00161, Rome, Italy.
authors.forename: A R
authors.initials: AR
authors.lastname: Ciccaglione
authors.affiliation: Department of Infectious, Parasitic and Immune-Mediated Diseases, Istituto Superiore di Sanità, Viale Regina Elena 299, 00161, Rome, Italy.
authors.forename: R
authors.initials: R
authors.lastname: Bruni
authors.affiliation: Department of Infectious, Parasitic and Immune-Mediated Diseases, Istituto Superiore di Sanità, Viale Regina Elena 299, 00161, Rome, Italy.
authors.forename: S
authors.initials: S
authors.lastname: Taffon
authors.affiliation: Department of Cell Biology and Neurosciences, Istituto Superiore di Sanità, Rome, Italy.
authors.forename: M
authors.initials: M
authors.lastname: Equestre
authors.affiliation: Laboratory of Transmissible Diseases and Biological Active Substances, Faculty of Pharmacy, University of Monastir, Monastir, Tunisia.
authors.forename: M
authors.initials: M
authors.lastname: Aouni
authors.affiliation: Department of Environment and Primary Prevention, Istituto Superiore di Sanità, Viale Regina Elena 299, 00161, Rome, Italy. giuseppina.larosa@iss.it.
authors.forename: G
authors.initials: G
authors.lastname: La Rosa
mesh_terms: D001483:Base Sequence; D004784:Environmental Monitoring; D017957:Hepatitis A Virus, Human / Q000145:classification / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D006801:Humans; D058889:Molecular Typing; D010802:Phylogeny; D036301:Qualitative Research; D012367:RNA, Viral / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D062211:Spatio-Temporal Analysis; D014416:Tunisia; D014504:Urban Health; D019562:Viral Load; D062065:Wastewater / Q000821:virology*; D018508:Water Purification
chemical_list: D012367:RNA, Viral; D062065:Wastewater
keywords: Genotype IA; Genotype IB; Hepatitis A; Nested PCR; Real-time PCR; Wastewaters