Browsing by Person "Mayr, Constanze F."

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Nachweis von luftgetragenen Viren an Standorten der Abfall- und Abwasserentsorgung
(2002) Mayr, Constanze F.; Boehm, Reinhard
In this study, aerosols workers at waste and wastewater treatment plants were exposed to, were examined with regard to the presence of airborne human pathogenic viruses. Besides the detection of hepatitis B virus and hantaviruses the main issue was to detect hepatitis A virus, rotavirus, human enteroviruses and poliovirus. Airborne viruses were sampled in parallel with different bioaerosol samplers, using various physical collection methods. At selected locations, measurements with two special-impingers, the MD 8-sampler and two high-volume-samplers were performed. Personal exposition of waste treatment workers to aerosols was measured using the PGP-GSP-system. Before viruses could be detected, the air samples had to be prepared. Because of the different sampling methods, several protocols had to be established. For the purification and concentration of the collected virus particles the following methods were used: prefiltration, ultrafiltration (100 and 500 KD), density gradient ultrafiltration using a sucrose gradient and the digestion of gelatine with trypsin. After these sampling processing procedures the virus detection and quantification of the samples of the special-impingers, the MD 8-sampler and the high-volume-samplers were assayed for human enteroviruses and rotavirus using four different tissue cultures (VERO, BGM, A549 and MA104) and molecular biological methods (RT-nested-PCR). For all other viruses only molecular biological methods (PCR) were applied. Regarding to the low flow-rate of the PGP-GSP-System only PCR was taken for virus detection. For the detection of viruses using the PCR assay, four different commercially available extraction-kits for DNA and RNA were compared. For all viruses besides hantavirus, several nested-PCR-systems for the qualitative and quantitative (using externe standards) determination with the LightCycler technique were established. To get informations about the attitude of a possible inhibition of the PCR reaction, the prepared samples were seeded with defined amounts of equine rhinovirus and amplified with an external standard on the LightCycler. The established PCR assays for hantavirus, hepatitis A virus, human enteroviruses and poliovirus showed a higher sensitivity (to be more sensitive) than the used cell culture. For detecting rotavirus, the sensitivity of both assays were equal, the detection limit for the nested-PCR of hepatitis B virus was one gencopy per reaction. In none of the 144 aerosol samples (45 PGP-GSP-samples and 99 from the other samplers) rotavirus, hepatitis A virus, hepatitis B virus and hantaviruses could be detected with the PCR technology. Also no rotavirus was isolated with the tissue culture method. In 21 cases the PCR showed positive results for human enterovirus RNA, the calculated concentration ranged from Äq. 100,47 KID50/ml to Äq. 103,74 KID50/ml. Viral infectivity could be demonstrated in 33 out of 99 examined specimens after inoculation on several cell cultures. The virus concentration ranged from 100,7 KID50/ml to more than 103,7 KID50/ml. After plaque purification some of the isolates were sequenced and identified as poliovirus type 1. In conclusion, the results of these studies indicate that at the investigated locations high measurable levels of hepatitis A virus, hantavirus, rotavirus and hepatitis B virus can not be expected. Nevertheless, in some special cases contamination might be possible. The detection of human enterovirus in 21 % of the location-based samples shows that especially bioaerosols at the waste water treatment plants might possibly be contaminated with viruses. Therefore the risk of infections can not be excluded.