Academic literature on the topic 'Specific F-RNA bacteriophages'

ABSTRACT The effect of natural inactivation in freshwater, chlorination, ammonia, extreme pHs, temperature, and salt content on phage inactivation was evaluated on mixtures of F-specific RNA bacteriophage isolates belonging to genotypes I, II, III, and IV. The bacteriophages studied were previously but recently isolated from natural samples, characterized as F-specific RNA bacteriophages and genotyped by plaque hybridization with genotype-specific probes. Natural inactivation in river water was modeled by in situ incubation of bacteriophages inside submerged dialysis tubes. After several days bacteriophages of genotype I showed the highest persistence, which was significantly different from that of bacteriophages of genotype II, IV, or III. The pattern of resistance of phages belonging to the various genotypes to extreme pHs, ammonia, temperature, salt concentration, and chlorination was similar. In all cases, phages of genotype I showed the highest persistence, followed by the phages of genotypes II, III, and IV. The phages of genotypes III and IV were the least resistant to all treatments, and resistance of genotypes III and IV to the treatments was similar. Bacteriophages of genotype II showed intermediate resistance to some of the treatments. The resistance of four phages of genotype I to natural inactivation and chlorination did not differ significantly. These results indicate that genotypes III and IV are much more sensitive to environmental stresses and to treatments than the other genotypes, especially than genotype I. This should be taken into consideration in future studies aimed at using genotypes of F-specific RNA bacteriophages to fingerprint the origin of fecal pollution.

An examination was made of the risk factors for gastrointestinal illness (GI) and other symptoms among canoeists and rafters using an artificial white-water canoe slalom course fed by a lowland river. The investigation was made by carrying out cohort studies of users on several days throughout one year. On each day water samples were collected for the determination of Escherichia coli, enterococci (faecal streptococci), F-specific RNA bacteriophage, sulphite reducing clostridia, culturable enteroviruses and turbidity. Of 755 questionnaires distributed, 473 (63%) were returned. The relative risks of GI and other symptoms were determined by logistic regression analyses. The variables associated with an increased risk of GI-illness were swallowing water, unintentional swimming in the course, eating and drinking before getting changed and the levels of F-specific RNA bacteriophages. Being a regular user was associated with a decreased risk of GI-illness. This study demonstrates the value of F-specific RNA bacteriophages as an index of risk from recreational use of a fresh water environment.

F specific RNA (FRNA) bacteriophages are attractive models for the study of the fate of animal viruses in water treatment processes and pure cultures (MS2, f2) have frequently been used in model experiments. The use of indigenous FRNA phages has been hampered by the lack of selective enumeration methods. Usually, bacteriophages in sewage are enumerated by counting the number of plaques produced in a lawn of an Escherichia coli host strain. Both somatic and F specific phages will contribute to the count. We report the development of a simple and highly selective method, based on a specially constructed host strain of Salmonella typhimurium (WG 49, phage type 3 Nalr (F'42 lac :: Tn5)). Counts on this host strain normally originate exclusively from F specific phages, because there are only few somatic Salmonella phages in most sewage samples. FRNA phage counts from natural sewage samples using this host strain are more reliable and higher than those obtained on E. coli hosts. F specific bacteriophages were detected in all sewage samples investigated in numbers ranging from 102 − 5 × 104 pfu/ml. Data are presented on the number and types of F specific phages in mixed domestic/industrial sewage; in poultry processing wastewater and in hospital wastewater. The potential of F specific phages as water quality indicators is discussed.

The occurrence of somatic (F') and male-specific (F') coliphages and Salmonella phages in a variety of environmental water samples was studied using different bacterial hosts. The number of plaque-forming units (pfu) of the different bacteriophages were compared and their resistance pattern to a biological treatment (humus tank) and chlorination was evaluated. The presence of the bacteriophages in shellfish was also studied. The morphology of isolate bacteriophages was examined as well as the visibility of the different plaques formed. Coliphages were found to produce larger and clearer plaques than all other bacteriophages studied. In most of the environmental water samples coliphages outnumbered all other bacteriophages, with the exception of dam water in which higher levels of F' Salmonella phages were detected. The majority of the F' Salmonella phages were shown to be RNA bacteriophages.

ABSTRACT In order to identify the origin of the fecal contamination observed in French estuaries, two library-independent microbial source tracking (MST) methods were selected: (i) Bacteroidales host-specific 16S rRNA gene markers and (ii) F-specific RNA bacteriophage genotyping. The specificity of the Bacteroidales markers was evaluated on human and animal (bovine, pig, sheep, and bird) feces. Two human-specific markers (HF183 and HF134), one ruminant-specific marker (CF193′), and one pig-specific marker (PF163) showed a high level of specificity (>90%). However, the data suggest that the proposed ruminant-specific CF128 marker would be better described as an animal marker, as it was observed in all bovine and sheep feces and 96% of pig feces. F RNA bacteriophages were detected in only 21% of individual fecal samples tested, in 60% of pig slurries, but in all sewage samples. Most detected F RNA bacteriophages were from genotypes II and III in sewage samples and from genotypes I and IV in bovine, pig, and bird feces and from pig slurries. Both MST methods were applied to 28 water samples collected from three watersheds at different times. Classification of water samples as subject to human, animal, or mixed fecal contamination was more frequent when using Bacteroidales markers (82.1% of water samples) than by bacteriophage genotyping (50%). The ability to classify a water sample increased with increasing Escherichia coli or enterococcus concentration. For the samples that could be classified by bacteriophage genotyping, 78% agreed with the classification obtained from Bacteroidales markers.

Difference in behaviors of F-specific DNA and RNA bacteriophages during coagulation–rapid sand filtration and coagulation–microfiltration (MF) processes were investigated by using river water spiked with F-specific DNA bacteriophage f1 and RNA bacteriophage f2. Because the particle characteristics of f1 (filamentous) and f2 (spherical) are quite different and the surface charge of f1 in the river water was slightly more negative than that of f2, the removal ratios of f1 were approximately 1-log lower than the removal ratio of f2 after any treatment process used in the present study. This result indicates that the behaviors of the two bacteriophages during the treatment processes were different, and that the removal of f1 by the combination of coagulation and filtration processes was more difficult than that of f2. The removal ratios for f1 and f2 were approximately 3-log and 4-log, respectively, in the coagulation–rapid sand filtration process, and 6-log and 7-log, respectively, in the coagulation–MF filtration process. Therefore, as expected, the coagulation–MF process appears to be more effective than the coagulation–rapid sand filtration process for the removal of not only spherical viruses but also filamentous viruses.

Male-specific (F) RNA bacteriophages have been proposed as indicators for human enteric viruses in shellfish. This study compared the use of Escherichia coli and FRNA bacteriophages to indicate the presence and level of noroviruses in Crassostrea gigas. A total of 167 samples from category A and B shellfish harvesting areas were analyzed for E. coli and FRNA bacteriophages by standard methods and for noroviruses (NoV) by using a previously described real-time PCR assay. FRNA bacteriophage and NoV levels in shellfish showed a seasonal trend and were elevated during the winter period (October through March). Conversely, E. coli levels did not reflect this seasonal trend. Categorizing samples on the basis of E. coli levels according to European Union regulatory limits failed to indicate the occurrence or level of NoV in shellfish. However, by grouping shellfish samples on the basis of FRNA bacteriophage levels a clear correlation was observed with NoV levels. The use of FRNA bacteriophages to predict the occurrence of NoV in shellfish could provide improved public health protection and should be considered when developing risk management procedures for shellfisheries.

A field study was performed to investigate reduction by dune infiltration and to estimate sticking efficiencies of F-specific RNA bacteriophages, total and thermotolerant coliforms, faecal streptococci and spores of sulphite-reducing clostridia. Reduction was considered as a β-binomially distributed process and a Monte Carlo simulation was applied for estimating sticking efficiencies. Reduction of F-specific RNA bacteriophages within the first 2m was 3.8 log10 and the sticking efficiency was about 0.002. The faecal indicator bacteria were removed only 0.9 log10 within 2m and sticking efficiency was 0.007. Concentrations of spores of sulphite reducing clostridia were reduced 1.9 log10 and their sticking efficiency was about 0.009.

Conjugative pili are extracellular filaments elaborated by Gram-negative bacteria expressing certain type IV secretion systems. They are required at the earliest stages of conjugal DNA transfer to establish specific and secure cell–cell contacts. Conjugative pili also serve as adsorption organelles for both RNA and DNA bacteriophages. Beyond these facts, the structure, formation and function of these filaments are poorly understood. This paper describes a rapid, quantitative assay for F-pili encoded by the F plasmid type IV secretion system. The assay is based on the specific lateral adsorption of icosahedral RNA bacteriophage R17 by F-pili. Bacteriophage particles conjugated with a fluorescent dye, Alexa 488, and bound to F-pili defined filaments visible by immunofluorescence microscopy. F-pili attached to F+ cells and free F-pili were both visible by this method. For quantification, cell-bound bacteriophage were separated from free bacteriophage particles by sedimentation and released by suspending cell pellets in 0·1 % SDS. Fluorescence in cell-free supernatant fractions was measured by fluorometry. The authors present a characterization of this assay and its application to F-pilus formation by cells carrying mutations in the gene for the F-pilus subunit F-pilin. Each mutation introduced a cysteine, which F-pilin normally lacks, at a different position in its primary structure. Cysteine residues in the N-terminal domain I abolished filament formation as measured by fluorescent R17 binding. This was confirmed by measurements of DNA donor activity and filamentous DNA bacteriophage infection. With one exception (G53C), cysteines elsewhere in the F-pilin primary structure did not abolish filament formation, although some mutations differentially affected F-pilus functions.

Current measures for controlling the public health risks associated with bivalve molluscan shellfish consumption rely on the use of Escherichia coli to indicate the sanitary quality of shellfish harvesting areas. However, it has been demonstrated that E. coli is an inadequate indicator of the viral risk associated with shellfish. An alternative indicator, male-specific B+ coliphages, have been investigated as viral indicators of faecal contamination that may provide source-specific information for impacted environmental waters. This study compared the distribution of E. coli and F+ RNA bacteriophages in shellfish grown in harvesting areas of Greece and also examined the presence and proportions of the different subgroups of F+ RNA coliphages in shellfish. F+ RNA bacteriophages were present in shellfish at higher concentrations than E. coli. Elevated numbers of F+ RNA bacteriophages observed in the winter concur with the known increased viral risk associated with shellfish harvested at that time of year in Greece. The majority of F+ RNA coliphages detected in shellfish samples belonged to group IV which indicated the possible presence of animal faecal material in sample harvesting areas. Phages of groups II and III (human waste and human faecal material, respectively) were present at low levels. Finally, 8% of the phages hybridised were found to belong to group I. The presence of group IV showed seasonal distribution (more in winter, less in summer) whereas the other groups did not show any difference. Monitoring of F+ coliphage subgroups may indicate the presence and major sources of microbial inputs to surface waters; however, environmental effects on the relative occurrence of different groups need to be considered.

Introduits dans l’environnement par l’intermédiaire de sources ponctuelles et diffuses, les virus et les bactériophages entériques peuvent se propager dans les cours d’eau par l’intermédiaire de différentes voies de dissémination. Détectées à la fois dans les eaux de surface et les sédiments des rivières, ces particules virales demeurent inertes dans le milieu hydrique. Leur propagation dépend donc uniquement des nombreuses interactions qu’elles partagent avec leur environnement. Qui plus est, la contamination virale des ressources en eau semble étroitement liée aux variations hydro-climatologiques. Mais malgré les connaissances déjà acquises à ce sujet, de multiples zones d’ombre subsistent concernant les variables et facteurs contrôlant le comportement in situ des particules virales dans le milieu hydrique. L’objectif de ce travail a donc été de définir le transport et le devenir des bactériophages ARN F-spécifiques dans une rivière en fonction de leurs caractéristiques propres et des conditions hydro-climatologiques. L’application de stratégies et méthodologies originales, tirées du domaine de l’hydrologie comme l’utilisation du temps de résidence de la masse d’eau ou l’échantillonnage automatique à haute fréquence, a permis d’étudier les comportements des bactériophages ARN F-spécifiques in situ. L’influence des facteurs environnementaux et plus particulièrement de la température de l’eau et du débit de la rivière sur la propagation et la survie in situ de ces particules infectieuses dans la colonne d’eau a été démontrée. Dans les sédiments, une distribution spatiale des bactériophages ARN F-spécifiques infectieux a été mise à jour. Cette particularité a pu être comprise et déchiffrée grâce à la combinaison de la caractérisation du sédiment et de l’étude du comportement d’attachement des quatre génogroupes. Les transferts de particules virales entre la colonne d’eau et les sédiments ont également pu être mis en exergue et s’avèrent être fortement dépendants des conditions hydro-climatologiques. Ainsi, la dynamique des bactériophages ARN F-spécifiques a pu être mieux appréhendée, et de même, les origines et la nature de la pollution virale ont été mieux discernées lors d’événements de crues. L’ensemble de ces résultats permet de compléter le puzzle de la dynamique des bactériophages ARN F-spécifiques dans la rivière. Les nouvelles approches expérimentales et méthodes d’analyse mises en place devraient permettre d’aboutir à une meilleure évaluation des risques viraux pour la santé humaine liés à l’utilisation des ressources en eau
Introduced into the environment through point and diffuse sources, enteric viruses and bacteriophages can be spread in watercourses via various dissemination routes. Detected in both surface water and river sediment, these viral particles remain inert in environmental water. Their spread is governed by many interactions that they have with their direct environment. Moreover, viral contamination of water resources is closely related to hydro-climatological variations. Despite the important knowledge already reported on this subject, many grey areas remain about the variables and factors controlling the in situ behavior of viral particles in environmental water. The aim of this study was therefore to define the transport and fate of F-specific RNA bacteriophages in a river according to their intrinsic characteristics and hydro-climatological conditions. The application of innovative strategies and methodologies from the hydrological science domain, such as the use of the residence time of the river water mass or high frequency automatic sampling, allowed studying the in situ behavior of F-specific RNA bacteriophages. The influence of environmental factors, especially water temperature and flow rate, has been demonstrated to have an impact on the in situ propagation and survival of infectious viral particles in the water column. Furthermore, the spatial distribution of infectious F-specific RNA bacteriophages was underlined in sediments. The accurate characterization of sediment and the study of the attachment capacity of the four genogroups explained this specific distribution. Finally, transfers of viral particles between the water column and sediment was highlighted and appeared to be highly dependent on hydro-climatological conditions. Besides the gained knowledge of the dynamics of F-specific RNA bacteriophages, the sources and origins of viral pollution of streams during rain and flood events were elucidated. This work helps completing the jigsaw puzzle on presence and transmission of F-specific RNA bacteriophages in river systems. The novel experimental approach further enhances human health-dependent viral risk evaluation linked to water resource utilization and management

Les virus entériques sont à l’origine de pathologies liées au péril fécal et dans l’état actuel des connaissances, la recherche des indicateurs de pollution fécale conventionnels (i.e. Escherichia coli, entérocoques) peut s’avérer inefficace pour évaluer le danger viral. La définition d’autres indicateurs pour gérer le danger lié à la présence des virus entériques dans les matrices hydriques et alimentaires est aujourd’hui nécessaire. Parmi eux, les bactériophages ARN F-spécifiques (FRNAPH) présentent plusieurs intérêts. Ces virus d’origine entérique sont présents en quantité importante dans les eaux usées. Très proches des virus entériques en termes de structure, ces microorganismes présentent l’avantage d’être facilement cultivables. Ils sont enfin souvent étudiés pour déterminer l’origine d’une pollution fécale (i.e. humaine ou animale). Certaines limites leur sont cependant fréquemment associées, que ce soit en termes de corrélation avec les pathogènes entériques ou concernant leur potentiel pour discriminer l’origine d’une pollution. Dans ce contexte, l’objectif du travail présenté ici était de préciser l’intérêt des FRNAPH en tant qu’indicateurs de pollution fécale mais aussi en tant qu’indicateurs de pollution virale dans l’environnement et les coquillages. Ces travaux ont permis dans un premier temps d’améliorer la capacité des FRNAPH à identifier les contaminations d’origine humaine. Nos résultats soulignent par ailleurs la plus-value apportée par la recherche des FRNAPH en cas de pollution fécale massive, en particulier si on s’intéresse à la contamination des coquillages. En effet, contrairement aux indicateurs bactériens, l’accumulation des FRNAPH ainsi que leur persistance dans ces aliments est très comparable à celles des virus entériques (i.e. norovirus). Enfin, en utilisant des méthodes de détection comparables, une forte corrélation entre la présence des FRNAPH d’origine humaine et celle des norovirus a été observée dans les coquillages. Compte tenu de ces résultats, une méthode de détection assurant la détection sensible des FRNAPH infectieux d’origine humaine dans différents types de matrices hydriques ou alimentaires (e.g. eaux de surface, fruits de mer, fruits rouges, salades) est proposée pour améliorer la gestion du danger viral
Enteric viruses are a leading cause of fecal-oral route transmitted diseases and currently, conventional fecal indicator bacteria (i.e. Escherichia coli, enterococcus) fail to assess this kind of hazard. In this context, the use of more efficient indicators to assess the hazard linked to viruses in water or foodstuff is required. F-specific RNA bacteriophages (FRNAPH) present numerous benefits for this purpose. Of enteric origin, these viruses are found in high concentrations in wastewater. Sharing many structural similarities with pathogenic enteric viruses, FRNAPH are easily cultivable and their potential to track the origin of the pollution is also often investigated. However, some limits are still associated with these indicators, regarding to their ability to track the origin of the pollution or concerning the lack of correlation with pathogens. In this context, the aim of this work was to make clear the potential of FRNAPH as fecal and as viral indicators in environmental waters and shellfish. As a first step, their ability to track human pollution was optimized. In addition, our results underlined the gains bringing by FRNAPH detection, especially when focusing on shellfish microbiological quality management. Indeed, unlike fecal indicator bacteria, the accumulation of FRNAPH and their persistence in shellfish have been found to be close to that of enteric viruses (i.e. norovirus). Furthermore, when using comparable methods for their detection, high correlation was observed between human FRNAPH and norovirus in shellfish. Taking into account these observations, a sensitive method allowing the detection of infectious FRNAPH of human origin was developed to improve viral hazard management in water and food commodities (e.g. environmental waters, shellfish, soft fruits, leaf)

Les masses d’eau douces sont sujettes à des contaminations fécales d’origines diverses. Parmi ces contaminants, les virus entériques, dont les norovirus (NoV), sont responsables de nombreuses épidémies à gastro entérites chaque année dans le monde. Les indicateurs de contamination fécale actuels (i.e. E. coli) recommandés par différentes réglementations s’avèrent peu fiables pour estimer le risque viral dans l’eau. D’autres indicateurs, ayant des caractéristiques proches des virus entériques, tels que les bactériophages ARN F-spécifiques (FRNAPH), sont proposés pour évaluer ce risque viral. Cependant, l’analyse des FRNAPH infectieux dans les eaux se heurte à certaines limites, notamment liées aux caractéristiques hydrodynamiques des milieux aquatiques. Afin de lever ces limites, une solution serait de réaliser les analyses au sein de capteurs accumulateurs et intégrateurs de ces cibles. Dans ce contexte, l’objectif de ce travail est de tester l’intérêt d’un mollusque bivalve dulcicole, la dreissène (Dreissena polymorpha), largement utilisé pour la surveillance chimique et écotoxique des masses d’eau, comme capteur biologique pour évaluer et suivre la contamination virale des masses d’eau. Pour ceci la stratégie suivie a consisté à i) caractériser la cinétique d’accumulation et de dépuration des FRNAPH infectieux chez la dreissène en conditions contrôlées de laboratoire et in situ, ii) définir un modèle toxico-cinétique pour formaliser la relation entre la concentration en FRNAPH infectieux dans la dreissène et le niveau d’exposition (concentration dans l’eau), iii) évaluer la contamination virale de masses d’eau sur une large échelle géographique et enfin iv) évaluer le couplage biocapteur-FRNAPH infectieux pour représenter la contamination des masses d’eau par le génome de NoV.Les données obtenues en laboratoire et in situ soulignent l’accumulation très rapide des FRNAPH infectieux par la dreissène avec une mise en équilibre avec son milieu en moins de 48h. De plus, les accumulations sont proportionnelles au niveau d’exposition sur une très large gamme de concentration et le signal en FRNAPH infectieux reste au sein des tissus de la dreissène plusieurs jours après l’exposition. Ainsi l’ensemble de ces données souligne l’intérêt de la dreissène comme système accumulateur et intégrateur. La définition d’un modèle toxico-cinétique à un compartiment, sur la base de ce qui est connu pour les contaminants chimiques, a permis de définir des facteurs de bioaccumulation in situ particulièrement intéressants (BCF ≈ 1 000) et autorisant un réel apport in situ. A l’aide d’une approche active (encagement d’organismes calibrés), le projet a validé l’apport de la dreissène comme biocapteur pour évaluer les concentrations en FRNAPH infectieux de nombreuses masses d’eau ainsi que son apport pour l’évaluation du risque viral vis à vis de la présence du génome de NoV
Freshwater bodies are subject to fecal contamination from a variety of sources. Among these contaminants, enteric viruses, including Noroviruses, are responsible for numerous gastroenteritis epidemics worldwide every year. The current fecal contamination indicators (i.e., E. coli) recommended by various regulations are proving unreliable for estimating the viral risk in water. Other indicators, with characteristics close to those of enteric viruses, such as specific-F RNA bacteriophages (FRNAPH), have been proposed to assess this viral risk. However, the analysis of infectious FRANPH in water comes up against certain limitations, notably linked to the hydrodynamic characteristics of aquatic environments. In order to overcome these limitations, one solution would be to carry out analyses using sensors that accumulate and integrate these targets. In this context, the aim of this work is to test the interest of a freshwater bivalve mollusc, the zebra mussel (Dreissena polymorpha), widely used for chemical and ecotoxic monitoring of water bodies, as a biological sensor for assessing and monitoring viral contamination of water bodies. The strategy followed consisted in i) characterizing the kinetics of accumulation and depuration of infectious FRNAPH in mussels under controlled laboratory and in situ conditions, ii) defining a toxico-kinetic model to formalize the relationship between the concentration of infectious FRNAPH in mussels and the level of exposure (concentration in water), iii) assess viral contamination of water bodies on a broad geographical scale, and finally iv) evaluate biosensor-infectious FRNAPH coupling to represent contamination of water bodies by the NoV genome.Data obtained in the laboratory and in situ underline the very rapid accumulation of infectious FRANPH by mussels, with equilibration with its environment in less than 48 hours. What's more, accumulations are proportional to the level of exposure over a very wide concentration range, and the infectious FRANPH signal remains in mussel tissues for several days after exposure. All these data underline the interest of D. polymorpha as an accumulator and integrator system. The definition of a single compartment toxicokinetic model, based on what is known for chemical contaminants, has enabled us to define particularly interesting in situ bioaccumulation factors (BCF ≈ 1,000) and authorizing a real in situ contribution. Using an active approach (caging of calibrated organisms), the project validated the contribution of zebra mussel as a biosensor for assessing infectious FRNAPH concentrations in numerous water bodies, as well as its contribution to viral risk assessment vis à vis the presence of the NoV genome