BGM Cell Lines Research Articles

Viral Interference as a Factor of False-Negative in the Infectious Adenovirus Detection Using Integrated Cell Culture-PCR with a BGM Cell Line

This study investigated the influence of viral interference on the detection of enteric viruses using the integrated cell culture (ICC)-PCR with a BGM cell line. It was possible to detect 102 plaque-forming units (PFU)/flask of enterovirus 71 (EV71) in spite of the presence of 104 PFU/flask of adenovirus 40 (AdV40). Meanwhile, 104 PFU/flask of AdV40 was not detected in the presence of 102 PFU/flask of EV71. This inhibition of AdV40 detection using ICC-PCR was attributable to the growth of EV71, because the addition of a growth inhibitor of EV71 (rupintrivir) neutralized the detection inhibition of AdV40. The growth inhibition of AdV40 under co-infection with EV71 is probably caused by the immune responses of EV71-infected cells. AdV is frequently used as a fecal contamination indicator of environmental water, but this study demonstrated that false-negative detection of infectious AdV using ICC-PCR could be caused by the co-existence of infectious EV in a water sample. The addition of rupintrivir could prevent false-negative detection of AdV using ICC-PCR. This study, therefore, emphasizes the importance of confirming the presence of multiple enteric viruses in a sample derived from environmental water prior to the application of ICC-PCR because the viral interference phenomenon may lead to the false-negative detection of target viruses.

THE STUDY OF CIRCULATION OF VIRUSES IN SURFACE WATERS AND IN WASTEWATER

Introduction. There was presented data from the field observation of surface waters and wastewater on indices of viral pollution. Material and Methods. To detect the presence of viruses, we examined samples as following: waters of the rivers of the Central European region of Russia; wastewater of the cities of this region; wastewater after treatment at three aeration stations. Eluates were examined for viruses on 2 transplantable RD and Hep-2 cell lines recommended by WHO, as well as on BGM culture. In addition, in the majority of samples, RNA of entero-, noro-, astro-, rotaviruses, hepatitis A viruses (HAV) was detected in RT-PCR and adenovirus DNA in PCR. The output of viruses in cell cultures was carried out in 3 passages, and the determination of RNA and DNA of viruses - in 2 replications. All samples were also examined for coliphages, as indirect indices of viral contamination. Results. It was revealed that coliphages were detected in surface water in 94% of samples, viruses on tissue RD, Hep-2 and BGM cell lines were detected in 35% of samples. The percentage of detection of RNA entero-, rota-, noro-, astroviruses, HAV and adenovirus DNA ranged from 10 to 70%. Discussion. Studies conducted at three aeration stations showed an increase in viral contamination of water in water bodies after the discharge of wastewater into them. Conclusion. Wastewater, both untreated and after biological treatment and disinfection, is contaminated with viruses and when discharged into surface water bodies creates a high risk to public health. In order to preserve the water resources of Russia, modern methods of cleaning and disinfecting wastewater should be modernized with the development of individual schemes taking into account the data of sanitary and virological control of discharged wastewater and the standards specified in the documents of water sanitary legislation.

Feasibility of the Bag-Mediated Filtration System for Environmental Surveillance of Poliovirus in Kenya

The bag-mediated filtration system (BMFS) was developed to facilitate poliovirus (PV) environmental surveillance, a supplement to acute flaccid paralysis surveillance in PV eradication efforts. From April to September 2015, environmental samples were collected from four sites in Nairobi, Kenya, and processed using two collection/concentration methodologies: BMFS (> 3 L filtered) and grab sample (1 L collected; 0.5 L concentrated) with two-phase separation. BMFS and two-phase samples were analyzed for PV by the standard World Health Organization poliovirus isolation algorithm followed by intratypic differentiation. BMFS samples were also analyzed by a cell culture independent real-time reverse transcription polymerase chain reaction (rRT-PCR) and an alternative cell culture method (integrated cell culture-rRT-PCR with PLC/PRF/5, L20B, and BGM cell lines). Sabin polioviruses were detected in a majority of samples using BMFS (37/42) and two-phase separation (32/42). There was statistically more frequent detection of Sabin-like PV type 3 in samples concentrated with BMFS (22/42) than by two-phase separation (14/42, p = 0.035), possibly due to greater effective volume assayed (870 mL vs. 150 mL). Despite this effective volume assayed, there was no statistical difference in Sabin-like PV type 1 and Sabin-like PV type 2 detection between these methods (9/42 vs. 8/42, p = 0.80 and 27/42 vs. 32/42, p = 0.18, respectively). This study demonstrated that BMFS can be used for PV environmental surveillance and established a feasible study design for future research.

Evaluation of novel pyrrolopyrimidine derivatives as antiviral against gastroenteric viral infections

Viral gastroenteritis is a major global public-health threat. All age groups are susceptible for this infection, but its most serious consequences affect children. Rotavirus, Coxsackievirus and Adenovirus are the most common viruses that cause gastroenteritis. Herein, we synthesized novel pyrrole, pyrrolo[2,3‑d]pyrimidine and pyrrolo[3,2‑e][1,2,4]triazolo[4,3‑c]pyrimidine derivatives. The non-toxic doses of these compounds were determined using BGM cell lines. We examined all the new compounds for their anti-viral activities against Rotavirus Wa strain and Coxsackievirus B4. Compounds 2a, 2d, 5a, 5c, 5d, 7b, 7j, 7n, 14b, 14c, 14e and 14f exhibited significant antiviral activity. We interpreted the action of these compounds using molecular docking against the homology models of viral polymerase enzymes of these viruses. RMSD value of 5d/Coxsackievirus was higher than the RMSD value for 5d/rotavirus and hence better as a stability parameter, which can be correlated to the biological activity.

Antiviral activities of some synthesized methylsulfanyltriazoloquinazoline derivatives

A series of some newly methylsulfanyl-triazoloquinazoline derivatives (1–21) have been tested against Coxsackievirus B4 and Adenovirus type 7. The antiviral activity of the titled compounds against Coxsackie B4 and Adeno type 7 was evaluated in BGM and Hep-2 cell lines. Compounds 4, 5, and 6 represent the most populated set of compounds obtained during this study, and 4 and 6 in particular were found to display the highest activities against Coxsackie B4 and exhibited good effect against Adeno type 7. The % of reduction in the case of Coxsackie B4 titer was 63.3 % (4, 6) and 83.3 % (5), while in Adeno type 7 titer was 63.3, 50, and 66.6 % for 4, 5, and 6, respectively.

Effect of Detection Methods on Risk Estimates of Exposure to Biosolids‐Associated Human Enteric Viruses

This study illustrates the effect of virus detection methods on estimates of risks of infection of biosolids-associated viruses for occupational workers and residential population during a hypothetical exposure of biosolids. Five gastroenteritis-associated human enteric viruses--enteroviruses (echovirus-12, enteroviruse types 68-71), adenoviruses, rotaviruses, and noroviruses genotype--I-were considered to represent human enteric viruses for risk estimation purposes. Ingested viral doses were calculated using literature-reported total infectious virus concentrations (based on BGM and A549 cell lines) and genome copies (GCs) in Michigan dewatered and class B biosolids. Cell-line-based infectivity parameters (i.e., ratio of total infectious virus concentration to GCs) were developed for different viruses in biosolids to use GCs for calculating ingested viral dose, addressing the issue of integration of molecular methods with biosolids-based virus risk assessment. Use of virus concentrations from molecular methods (with and without using cell-line-based infectivity parameter) resulted in higher risk estimates than culture methods, indicating the effect of the virus detection method on risk estimates. Further, use of virus concentrations from A549 cell lines resulted in higher risk estimates compared to those from BGM cell lines, suggesting the need for a proper choice of cell lines in determining infectious viral dose. The Monte Carlo uncertainty analyses of estimates for risk of infection due to enteroviruses showed that enteroviruses concentration was the most important parameter influencing risk estimates, indicating the need for reducing associated uncertainty. More work is required to develop cell-line-based infectivity parameters for different virus concentration levels and sample matrix types using a cut-off-based approach.

A comparison of BGM and LLC-PK1 cells for the evaluation of nephrotoxicity

Nephrotoxicity is one of the most frequent effects observed after the use of medicine. Such situations have been tardily discovered because of existing methods to determine toxicity. The validation of sensitive, alternative methods for the early identification of toxic effects is as important as restrictions on the use of animals. In this light, the present study evaluated the effects of gentamicin on BGM and LLC-PK1 cells, using MTT and Neutral Red (NR). Although the LLC-PK1 cell line is used for toxicological studies, the BGM cell line is relatively new for this purpose. MTT (BGM: EC50 = 6.29 mM; LLC-PK1: EC50 = 8.01 mM) was found to be more sensitive than NR (EC50 was greater than 10 mM for both cells). By using MTT, both cells demonstrated the involvement of mitochondria in a manner that was dose dependent, with an apoptotic process occurring at the concentrations of 1 and 3 mM and necrosis at concentrations above 4 mM. It could, therefore, be concluded that 1) BGM appears to be useful in the study of the mechanism of nephrotoxicity caused by gentamicin and 2) because of its sensitivity to MTT, in addition to its ease of manipulation, it is believed that the BGM cell line can also be used as an alternative method to evaluate nephrotoxicity.

Synthesis and antimalarial activity of thioetherhydroxyethylsulfonamides, potential aspartyl protease inhibitors, Part 3

A series of novel thioetherhydroxyethylsulfonamide derivatives has been synthesized from the coupling of intermediates 3-amino-4-phenyl-1-thioetherazine-butan-2-oles 6,7 with arenesulfonyl chlorides in good yields. Characterizations of products were achieved by NMR techniques and specifically for compound 8e by X-ray crystallography. Preliminary results of antimalarial activity in vitro against the Plasmodium falciparum W2 clone (chloroquine resistant and mefloquine sensitive) showed moderate activity for hydroxyethylsulfonamide 8f. In addition, none of the compounds tested showed cytotoxicity at high concentration tested against HepG2 and BGM cell lines.

Comparison of BGM and PLC/PRC/5 Cell Lines for Total Culturable Viral Assay of Treated Sewage

The objective of this study was to compare PLC/PRF/5 and BGM cell lines for use in a total culturable viral assay (TCVA) of treated sewage effluents. Samples were collected before and after chlorination from an activated sludge wastewater treatment plant and from the effluent of a high-rate enhanced flocculation system, followed by UV light disinfection. Cell monolayers were observed for cytopathic effect (CPE) after two passages of 14 days each. Monolayers exhibiting viral CPE were tested for the presence of adenoviruses and enteroviruses by PCR or reverse transcription-PCR. Eight percent of the samples exhibited CPE on BGM cells, and 57% showed CPE on PLC/PRF/5 cells. Only enteroviruses were detected on the BGM cells, while 30% and 52% of the samples were positive for enteroviruses and adenoviruses, respectively, on the PLC/PRF/5 cells. Thirty percent of the samples were positive for both adenoviruses and enteroviruses in chlorinated activated sludge effluent. Thirty percent of the samples were positive for adenoviruses in the UV treatment effluent, but no enteroviruses were detected. In conclusion, the PLC/PRF/5 cells were more susceptible than BGM cells to viruses found in treated sewage. The use of BGM cells for TCVA may underestimate viral concentration in sewage effluent samples. The PLC/PRF/5 cells were more susceptible to adenoviruses, which is important in the evaluation of UV disinfection systems because adenoviruses are highly resistant to UV inactivation.

Study on Detection, Isolation and Identifications of Infectious Viruses in Various Water Samples

Various factors using cell lines can effect kinds and frequencies of infectious viruses obtained in the detection tests on various water samples. We tried to find out technical problems for the maximum virus isolations from water samples and characterize the virus isolates from waters in nature and in various purification stages. Fourteen viruses were isolated from 169 water samples by virus monitoring protocol for the information collection requirements rule, US EPA. The morphological changes caused by viruses and mycoplasma infections were compared with for increasing the specificity of tests employed. Cytopathic effects of slow growing viruses were found very similar with those by toxic effects in water samples and mycoplasma infections. Five of 6 stream water samples tested (83.33%) showed virus contaminations with the range of 1.03 to 5.75 MPNs/100 liter. Eight of 24 source water samples (33.35%) showed viral contaminations. One water sample of 24 water samples during precipitation stages was shown to include infectious viruses. It was confirmed that infectious viruses were significantly decreased by purification stages from streams. The titers (TCID??) of virus isolates were ranged as 10 -6.8 ~ 10 -6.925 /㎖. The virus isolates were identified by immune fluorescent antibody (IFA) method using virus specific immune sera and serotyped using serotype specific reference sera. Of 14 virus isolates, 7 samples were identified as poliovirus and the other 7 were identified as coxsakie virus. Of 7 polioviruses, one was serotyped as type I, 3 viruses as type II and another 3 as type Ⅲ. Conclusively, BGM cell lines must be free of mycoplasma for the strict examination of infectious viruses in water and highly sensitive for mainly enteroviruses. In addition, most of infectious viruses showing typical cytopathic effect from water samples were confirmed as coxsackie B and live attenuated vaccine strains of 3 polio types when BGM cells were used for virus isolations.

Comparison of multiple shell vial cell lines for isolation of enteroviruses: A national perspective

Background Isolation of enterovirus in cell culture is still utilized by clinical laboratories, for vaccine research, and for identifying serotypes for disease surveillance. Use of a combination of cell lines is recommended yet this practice has not been rigorously examined for shell vials. Objectives To evaluate the growth predilection of enterovirus serotypes for certain shell vial cell lines in clinical samples received at a national reference laboratory. Study design We retrospectively reviewed results of samples submitted for viral culture over a 3-year period. PMK, BGM, RD, A549, and MRC-5 cell lines grown in shell vials were inoculated and serotyped. Results Of 55,816 cultures, 1047 (1.9%) yielded enterovirus representing 18 serotypes. Echovirus 7, echovirus 9, and echovirus 30 were the most common serotypes recovered in 2002, 2003, and 2004, respectively. PMK and MRC-5 recovered the majority of enterovirus isolates; the addition of BGM and RD cells increased our recovery rate by 13%. For 52.6% of enteroviral isolates, cytopathic effect was found in only a single cell line. PMK and BGM cells were effective in isolating coxsackieviruses, and RD and MRC-5 were useful particularly in isolating echoviruses. Conclusions A combination of shell vial cell lines is still recommended for recovery of enteroviruses.

Taura Syndrome Virus and Mammalian Cell Lines

Taura Syndrome Virus and Mammalian Cell Lines

Dynamics of the development of Chlamydophila psittaci inclusions in epithelial and fibroblast host cells.

The development of Chlamydophila psittaci (formerly Chlamydia psittaci, avian strains) inclusions in fibroblast L-929 and epithelial BGM cell lines was studied along the bacterial growth cycle using a BGM cell-adapted strain in the presence or absence of cycloheximide and cycloheximide + polyethylene glycol. Evolution of the inclusions was determined in terms of their number and size at 24, 30, 36, 48 and 54 h after infection. Significant differences in the chlamydial growth were found between both host cells, throughout the study. Higher numbers of inclusions (P < 0.05) were observed in L cells while larger inclusions (P < 0.01) were found in BGM cells. In both fibroblast and epithelial cells, inclusions showed a significant (P < 0.001) increase in size at the later times studied. Free extracellular chlamydial particles were noticed at 48 and 54 h post-infection (p.i.) in infected L cells, and at 54 h p.i. in BGM cells. Addition of cycloheximide or cycloheximide + polyethylene glycol had no significant effect on the number of inclusions or their size. The results suggest that host cell characteristics and innate compatibility between Chlamydophila strain and host cell are more important than host cell adaptation for the development of the microorganism.

Elimination of enteroviruses, other enteric viruses, F-specific coliphages, somatic coliphages and E. coli in four sewage treatment plants of southern Germany

Research Article| May 01 2000 Elimination of enteroviruses, other enteric viruses, F-specific coliphages, somatic coliphages and E. coli in four sewage treatment plants of southern Germany Jens Fleischer; Jens Fleischer 1Landesgesundheitsamt Baden-Württemberg, Wiederholdstr. 13, Infektiologie, D-70174 Stuttgart, Germany Search for other works by this author on: This Site PubMed Google Scholar Karin Schlafmann; Karin Schlafmann 2Universitätsklinikum Tübingen, Hygiene-Institut, Abt. für Allgemeine Hygiene und Umwelthygiene, Wilhelmstr. 31, D-72074 Tübingen, Germany Search for other works by this author on: This Site PubMed Google Scholar Rowland Otchwemah; Rowland Otchwemah 2Universitätsklinikum Tübingen, Hygiene-Institut, Abt. für Allgemeine Hygiene und Umwelthygiene, Wilhelmstr. 31, D-72074 Tübingen, Germany Search for other works by this author on: This Site PubMed Google Scholar Konrad Botzenhart Konrad Botzenhart 2Universitätsklinikum Tübingen, Hygiene-Institut, Abt. für Allgemeine Hygiene und Umwelthygiene, Wilhelmstr. 31, D-72074 Tübingen, Germany Search for other works by this author on: This Site PubMed Google Scholar Journal of Water Supply: Research and Technology-Aqua (2000) 49 (3): 127–138. https://doi.org/10.2166/aqua.2000.0011 Views Icon Views Article contents Figures & tables Video Audio Supplementary Data Share Icon Share Twitter LinkedIn Tools Icon Tools Cite Icon Cite Permissions Search Site Search nav search search input Search input auto suggest search filter All ContentAll JournalsThis Journal Search Advanced Search Citation Jens Fleischer, Karin Schlafmann, Rowland Otchwemah, Konrad Botzenhart; Elimination of enteroviruses, other enteric viruses, F-specific coliphages, somatic coliphages and E. coli in four sewage treatment plants of southern Germany. Journal of Water Supply: Research and Technology-Aqua 1 May 2000; 49 (3): 127–138. doi: https://doi.org/10.2166/aqua.2000.0011 Download citation file: Ris (Zotero) Reference Manager EasyBib Bookends Mendeley Papers EndNote RefWorks BibTex The reduction processes at four advanced sewage treatment plants in Baden-Württemberg were evaluated with regard to virus elimination and the elimination of indicator organisms from wastewater. The results of virus elimination were compared with the reduction of somatic and male specific bacteriophages and of E. coli. In total, 222 water samples were examined. The results obtained for the different treatment plants show reduction rates from 80.0% to 99.9% for enteroviruses, enumerated as PFU l−1 on BGM cell line, and reduction rates from 59.4% to 99.9% for other enteric viruses, enumerated as MPN l−1 on MA-104 cell line. Identification of the isolated enteroviruses yielded 88.3% for Coxsackie virus B (1-5), 18.3% were positive for Polio (1-3) and 8.3% for Echo virus (1+11). The reduction rates of somatic bacteriophages ranged from 76.4% to 99.90%, for male specific bacteriophages from 87.5% to 99.9% and for E. coli. from 75.0% to 99.9% respectively. Two of the plants use standard chemical precipitation and the other two employ combinations of chemical and biological elimination techniques to reduce the concentrations of phosphorus and nitrogen. A correlation between the amount of precipitators and the elimination rates of the tested microorganisms could not be demonstrated, perhaps due to the fact that the treatment conditions could not be modified by the investigators. It is concluded that the tested treatment plants using combinations of chemical and biological techniques for P and N removal show equal or higher elimination rates than conventional treatment processes using chemical elimination techniques. biological phosphorus elimination, coliphages, E. coli, sewage treatment, virus adsorption, virus elimination This content is only available as a PDF. © IWA Publishing 2000 You do not currently have access to this content.

F-specific RNA bacteriophages are adequate model organisms for enteric viruses in fresh water

Culturable enteroviruses were detected by applying concentration techniques and by inoculating the concentrates on the BGM cell line. Samples were obtained from a wide variety of environments, including raw sewage, secondary effluent, coagulated effluent, chlorinated and UV-irradiated effluents, river water, coagulated river water, and lake water. The virus concentrations varied widely between 0.001 and 570/liter. The same cell line also supported growth of reoviruses, which were abundant in winter (up to 95% of the viruses detected) and scarce in summer (less than 15%). The concentrations of three groups of model organisms in relation to virus concentrations were also studied. The concentrations of bacteria (thermotolerant coliforms and fecal streptococci) were significantly correlated with virus concentrations in river water and coagulated secondary effluent, but were relatively low in disinfected effluents and relatively high in surface water open to nonhuman fecal pollution. The concentrations of F-specific RNA bacteriophages (FRNA phages) were highly correlated with virus concentrations in all environments studied except raw and biologically treated sewage. Numerical relationships were consistent over the whole range of environments; the regression equations for FRNA phages on viruses in river water and lake water were statistically equivalent. These relationships support the possibility that enteric virus concentrations can be predicted from FRNA phage data.

Comparison of Cell Culture and a Poliovirus Gene Probe Assay for the Detection of Enteroviruses in Environmental Water Samples

Nucleic acid hybridization provides a rapid non-cell culture method for the detection of enteric viruses in water. The purpose of this work was to compare the detection of naturally occurring enteroviruses by cell culture with their detection by a poliovirus gene probe in various types of water samples. Samples of activated sludge effluent, tertiary treated wastewater (activated sludge, filtration and passage through reverse osmosis), ground water, surface water and tidal river water were processed through 1 MDS Virozorb filters to concentrate any naturally occurring virus. Viruses were eluted from the filters with pH 9.5 beef extract and reduced in volume by flocculation to 20-30 ml. These concentrates were then assayed in the BGM cell line by the cytopathogenic effects (CPE) method and by a poliovirus cDNA probe (base pairs 115-7440) labeled with 32P. A total of 233 samples were assayed in this manner. In slightly more than 93% of the samples gene probe and cell culture yielded the same results. Of these samples 36 were positive by gene probe and 28 by cell culture assay. Positive samples for gene probe were confirmed by treatment with NaOH or RNAse and then reprobed. Samples demonstrating CPE upon primary passage were confirmed positive by subsequent passage of cell lysate on a new monolayer of BGM cells. Ten samples were positive by gene probe and negative by cell culture, and 4 samples were negative by gene probe and positive by cell culture.

Processing procedures for recovering enteric viruses from wastewater sludges

A powdered beef extract specially formulated for recovering viruses from environmental samples and designated as beef extract V was evaluated using indigenous and viral seeded wastewater sludge samples. When beef extract V was used to process activated and aerobically digested sludge solids, virus recoveries were shown to be similar to other methods that used commercially available supplemented beef extract. When used to process other sludge solids (primary and activated without primary clarification), cytotoxicity resulted in the BGM cell line used for virus assay. When these sludge solids were processed with the supplemented commercially marketed beef extract cell toxicity did not occur. Metal concentrations in the processed sewage sludge eluates were analyzed, but based on the levels observed they could not be shown as the source of the cytotoxicity. This did not exclude possible synergistic cytotoxic effects or organometal complexes. The commercially marketed beef extract was supplemented with either a floccing aid (FeCl 3), a filter aid (Celite) or a floc prepared from paste beef extract. The paste beef floc supplement proved to be the most useful and reliable method for processing for viruses from wastewater sludge solids.

Microbiological Methods for Safety Testing of Drinking Water Directly Reclaimed from Wastewater

The following results were obtained in research on microbiological methods for assessment of the safety of drinking water directly reclaimed from wastewater: Studies on a wide variety of pathogenic micro-organisms revealed that the safety of the water could reliably be monitored by practical indicators such as standard plate counts, coliform bacteria, coliphages and acid-fast bacteria. A convenient plate incorporation modification of the Ames Salmonella mutagenicity assay proved suitable for routine quality surveillance of potentially mutagenic and carcinogenic compounds; results were confirmed by comparative tests using a hamster embryonic cell transformation assay in which sensitivity was increased by preparing growth media from test water. Screening for toxic activity was reliably done by assays based on interference with the multiplication of Pseudomonas putida, the cloning efficiency of BGM, HeLa and mouse lymphoma cell lines, and activity of the enzyme urease. A commercial limulus amoebocyte lysate assay proved a reliable and sensitive method for the detection of endotoxins. Correlation with results obtained in sophisticated chemical analyses, as well as long-term epidemiological studies on consumers, confirmed that, subject to the maintenance of certain operational conditions, the safety of directly reclaimed drinking water could reliably be monitored by relatively simple, inexpensive and rapid microbiological tests and bioassays.

Comparison of fortified calf serum, serum substitutes and fetal calf serum with or without extenders for propagation of cell cultures for virus plaque assays

Two studies comparing sewage-isolated and laboratory stock viruses were conducted to determine if alternative forms of serum or serum extenders could be used in place of fetal bovine serum without a significant loss of viral titer. In the first study, BGM cells were grown in standard MEM-L15 medium which was supplemented with Nuserum, Sigma serum replacement (CPSR-1), HyClone defined iron supplemented calf bovine serum, fetal bovine serum (FBS) or FBS supplemented with either SerXtend or Mito serum extenders. Comparison of virus titers showed that CPSR-1 gave the best overall results and was comparable to FBS. Of the serum extenders, only SerXtend improved virus recovery from sewage samples. In the second study, all sera were tested with and without SerXtend. In these experiments, SerXtend enhanced virus sensitivity of the BGM cell line grown in the HyClone serum but reduced the sensitivity of those cultured in Sigma serum. In both series, the growth of BGM cells was monitored for 12 weeks and all test products were shown to support long-term cell growth.

Comparison of cytopathogenicity, immunofluorescence and In situ DNA hybridization as methods for the detection of adenoviruses

Three different methods were compared for their efficiency at detection of adenoviruses. The samples examined for viral analysis consisted of concentrates prepared from raw sewage, chosen as providing a representation of the spectrum of viruses being intestinally shed from a large population at any given time. When using one single cell line, HEp-2, the overall numbers of adenoviruses detected using cytopathogenicity and immunofluorescence were roughly equal. In situ hybridization was approx. 40% more sensitive than either of these other methods as determined by average virus titers for the different samples, and also proved to be better by means of a nonparametric comparison. The 293 cell line was approx. 5 times more sensitive for detecting adenoviruses by cytopathogenicity as compared with the HEp-2 cell line, but proved unsuitable in our hands for quantitatively detecting indigenous adenoviruses by immunofluorescence. The relative number of indigenous adenoviruses present in the sewage concentrates we examined was, on average, 94-fold greater than that of enteroviruses. Assay of enteroviruses was performed by plaque assay in the BGM cell line.