[Microorganisms surviving in drinking water systems and related problems]. / Sopravvivenza microbica nelle reti di distribuzione di acque potabili e problemi conseguenti.

Drinking water in distribution systems may show abnormal values of some parameters, such as turbidity, and may support particular phenomena, such as bacterial regrowth or presence of Viable Not Culturable (VNC) bacteria. Turbidity can provide shelter for opportunistic microorganisms and pathogens. The Milwaukee outbreak (400,000 people) is one example of waterborne disease caused by the presence of pathogens (Cryptosporidium) in drinking water characterized by high and intermittent levels of turbidity. Bacterial regrowth in drinking water distribution systems may cause high increments of microorganisms such as heterotrophic bacteria, coliforms and pathogens. Microorganisms isolated from biofilm including Pseudomonas, Aeromonas, Legionella may have a significant health hazard especially in hospital areas. The presence of VNC bacteria in drinking water may represent a problem for their discussed role in infectious diseases, but also for the possibility of a considerable underestimation of true microbial concentrations in drinking waters. To study this kind of problems is necessary to apply suitable methods for drinking water analyses.

Microbial quality of wastewater: detection of hepatitis A virus by reverse transcriptase-polymerase chain reaction.

AIMS: The persistent circulation of hepatitis A virus (HAV) in the Mediterranean area suggests the need for monitoring its presence in the environment. A reverse transcriptase-polymerase chain reaction (RT-PCR) was used to detect the presence of HAV in several consecutive raw sewage and final effluent samples, collected over an 8-month period from an activated sludge treatment plant in southern Italy. METHODS AND RESULTS: Two distinct purification protocols, either based on antigen-capture with monoclonal antibody (AC) or RNA extraction, were compared. The possible influence of the antibody used in the AC phase was evaluated in preliminary experiments on HAV-spiked samples, using two different monoclonal antibodies. Hepatitis A virus RNA was detected in all but one sewage environmental sample examined. The contemporary presence of enteroviruses, reoviruses and phages was observed, while HAV growth in cell culture was hampered. CONCLUSIONS: The RT-PCR technique was confirmed to be a valuable tool for the rapid monitoring of HAV in sewage samples. In addition, this study demonstrated that application of different sample purification methods can result in different levels of sensitivity of the assay and that, in the antigen-capture method, the choice of antibody can have a crucial role. SIGNIFICANCE AND IMPACT OF THE STUDY: This work underlines the need for technical uniformity in environmental studies from different laboratories for a correct and useful comparison of the results.

Bacteriological and virological quality of seawater bathing areas along the Tyrrhenian coast.

Monitoring was carried out during summer 1997 along a selected area of the Tyrrhenian coast near the Tiber river mouth. Fifty-eight seawater samples, collected from 19 stations, were examined for coliforms, streptococci, Enteroviruses, Salmonellae, coliphages, Bacteroides fragilis phages, Pseudomonas, alophilic Vibrios, Aeromonas and yeasts. Salmonellae and coliphages were isolated in 3 and 12 out of 58 samples, respectively. Enteroviruses and Bacteroides fragilis phages were not isolated. Reoviruses were isolated only from 2 out of 58 samples. A limited number of samples of the northern stations located near the Tiber and other river mouths exceeded the guide values for bathing water by the EU Directive. All the southern stations, located near canals, were of very good microbiological quality. Pseudomonas, Vibrio, Aeromonas and yeasts were isolated from all stations and their values in 100 ml of seawater were 10-10(6), 10-10(6), 0-10(6) and 1-10(3), respectively. An extensive disinfection practice carried out on domestic wastes, which are discharged in rivers and canals, probably brought pollution levels of most stations to values within the bacterial standards. The spread of Pseudomonas, Aeromonas, etc. showed that all the coastal area studied was characterized by the presence of organic matter coming from land that can support the presence of opportunistic pathogens and other microbial flora.

[Microbiological quality of the Catania coastal sea water]. / La qualità microbiologica delle acque marine del litorale di Catania.

This study was carried out from 1997 to 1998 along a selected coastal area near Catania to ascertain bacteriological and virological quality of marine waters. 44 seawater samples, collected from 4 stations, were assayed for the presence of total and fecal coliforms, fecal streptococci, coliphages, Salmonellae and enteric viruses. Two stations localized at canal outfalls showed high levels of fecal pollution. The other stations were of good microbiological quality and showed a limited number of samples exceeding the standards laid down as guide values for bathing waters by Italian normative during the bathing period. Salmonellae were isolated in 8 out of 44 sea water samples (18%). Their presence was ascertained mainly in samples of the two polluted stations. Enteroviruses were not isolated. Enteric viruses such as Reoviruses were isolated from all stations, in 12 out of 44 samples (27%). The presence of these viruses was ascertained only during autumnal and winter seasons. The results of this study showed that, notwithstanding some stations showed high levels of bacteriological indicators of fecal pollution and presence of Salmonellae, enteroviruses growing on cell cultures were not isolated. Reoviruses confirmed their high diffusion in marine waters.

Identification of enteroviruses isolated from sea-water: indirect immunofluorescence (IIF).

In this research the Enteroviruses presence in sea water was studied. In previous studies presumptive Enteroviruses were revealed in 43% and in 77% of sea water samples analyzed. It was necessary to identify viral particles isolated from marine water because the detection of this kind of virus was performed only on the basis of cytopathic effect appearance on cell cultures. The aim of this research was to verify the suitability of Indirect Immunofluorescence for identification of presumptive Enteroviruses isolated from marine waters. 13 field strains from RC 37 cells (Cercopithecus Kidney cell line) were tested. Pools of Horse immune sera against COXS, POLIO and ECHO, and anti-horse antibodies conjugated with fluorescein were used. The results revealed the presence of Coxsackie virus and Echo virus. There were many problems related with ECHO identification for the presence of cross-reactions with COXS. The IIF method cannot be performed routinely, because it did not show high level of specificity and sensibility, and it is expensive. This methodology can be used only in particular cases and in laboratories with competent virologists and related facilities. In the future, the only suitable methodology for identification of Enteroviruses isolated from field samples is based on the use of RNA and DNA probes.

[Microbiological profile in water-supply conduits from a clogged well]. / Rilevamenti microbiologici in condotte di rifornimento idrico di un pozzo occluso.

This article presents the results of an investigation involving bacterioflora in a water well clogged for the presence of biomass. The water well, placed in a zone near Rome, showed some problems about the water quality and about the extraction of water. The examination of the interior of the pipes showed the presence of biomass. The biomass was examined microscopically and bacteriological analyses were carried out on it. Heterotrophic bacteria were enumerated with three different media by direct count, Pseudomonas sp., yeasts and fungi also by spread plate method. The anaerobic Sulphate Reducing Bacteria were investigated by "Most Probable Number" technique. The results of the analyses showed the presence of protozoa and algae. Moreover high quantity of bacterial flora as heterotrophic bacteria and Pseudomonas sp. were revealed. Sulphate Reducing Bacteria were enumerated in low quantities. Sphaerotilus natans, Actinomyces and Rhodotorula were identified. The clogging problems arose from the presence of filamentous microorganisms as Sphaeroilus natans and Actinomyces sp. When microorganisms of this kind are present in aquifers they can multiply massively if the conditions are favorable.