Paul A. Rochelle's Books

Comparing Cell Culture and Mouse Assays for Measuring Infectivity of Cryptosporidium

By: Paul A. Rochelle,Marilyn M. Marshall,J. Mead,JS Rosen

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Comparing Cell Culture and Mouse Assays for Measuring Infectivity of Cryptosporidium

By: Paul A. Rochelle,Marilyn M. Marshall,J. Mead,JS Rosen

Cell culture techniques are routinely used for measuring the infectivity of a wide range of human pathogens. A variety of different cell culture systems and detection methodologies have been applied to Cryptosporidium parvum. However, the correlation between cell culture methods and animal infectivity assays has not been thoroughly investigated. Although many cell culture methods have been developed for C. parvum, it has not been proven that infectivity in cell culture is a good indicator of the ability of oocysts to cause infections in animals. The objective of this research was to compare in-vitro cell culture methods with a mouse assay for measuring infectivity of C. parvum oocysts. The specific objectives were to (1) compare the dose response and sensitivity of cell culture and mouse assays with multiple isolates; (2) compare infectivity methods with oocysts exposed to environmental water samples; (3) determine the reproducibility and variability of the methods; and (4) compare cell culture and animal assays for assessing ozone and UV disinfection.For untreated oocysts, challenge doses were enumerated by flow cytometry. Dose response curves were constructed by regression analysis of oocyst dose against a logistic transformation of the proportional infectivity and the 50% infectious doses for each isolate were calculated by solving the regression for a logit value of zero. Infections in CD-1 mice were detected by microscopy following staining with hematoxylin and eosin. Infection in HCT-8 and Caco-2 cells was detected by C. parvum-specific RT-PCR. In MDCK cells, infection was detected using immunofluorescence. For disinfection studies, oocysts were exposed to UV using a medium-pressure, collimated beam apparatus and inactivation was measured as the difference in ID50 of unexposed and UV-exposed oocysts. Oocysts were exposed to ozone using batch, semi-batch, and single continuously stirred tank reactors at 1, 5, and 15°C.This investigation demonstrated that in-vitro cell culture was equivalent with a mouse assay for measuring infectivity of untreated C. parvum oocysts and should therefore be considered a practical alternative for assessing the potential of oocysts to cause infection. However, the high levels of variability displayed by mouse and cell culture methods indicated that infectivity and disinfection experiments should be limited to discerning relatively large differences. Of the three cell culture assays, the HCT-8/RT-PCR method displayed the closest agreement with the CD-1 mouse assay. C. hominis was infectious in HCT-8 cells but did not infect mice. Similar results were obtained with CD-1 mice and HCT-8 cells for measuring infectivity of oocysts that had been exposed to environmental water for 35 days. There was also very good agreement between HCT-8 cell culture and CD-1 mouse assays for measuring UV inactivation of C. parvum. A medium-pressure UV dosage of 5.6 mJ/cm2 resulted in 2-log10 inactivation. The shapes of ozone inactivation curves were generally the same for mouse and cell culture derived data although the CD-1 mouse assay typically generated 0.5 to 1-log10 higher levels of inactivation than HCT-8 cells. In addition, there was a stimulatory response in oocysts exposed to ozone below 20 mg.min/L when assayed by HCT-8 cell culture. Consequently, further research is necessary to understand the response of oocysts to ozone when inactivation is assessed by cell culture methods. The water industry should adopt in-vitro cell culture as a routine method for measuring the infectivity of waterborne C. parvum and C. hominis oocysts. This project has demonstrated that cell culture has equivalency with the standard CD-1 mouse assay and cell culture assays can be applied to oocysts recovered from water using approved methods. However, there needs to be a thorough, robust, and well-controlled study to compare the various cell culture-based assays for measuring C. parvum and C. hominis infectivity. This evaluation should include inter-laboratory comparisons and round-robin testing. Cell culture-based assays should also be used to assess disinfection of C. hominis isolates. Originally published by AwwaRF for its subscribers in 2004. This publication can also be purchased and downloaded via Pay Per View on Water Intelligence Online - click on the Pay Per View icon below

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Page Count

136

Publisher

IWA Publishing

Published Date

ISBN 10

1843399156

ISBN 13

9781843399155

Evaluation of Genotyping Techniques for Crytosporidium Parvum

By: Paul A. Rochelle,Ricardo De Leon,Edward R. Atwill

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Evaluation of Genotyping Techniques for Crytosporidium Parvum

By: Paul A. Rochelle,Ricardo De Leon,Edward R. Atwill

A research teams reports on its efforts to develop and implement DNA fingerprinting techniques to characterize and differentiate the parvus species of protozoal parasite that is ubiquitous in rivers and lakes, is relatively resistant to chlorine disinfection at commonly used concentrations, and can prove fatal for immune compromised people who become infected. They evaluate fingerprinting methods based on polymerase chain reaction, determine which method is most suitable for determining strains or isolates within a species, implement the most appropriate molecular method for determining whether the isolates can be differentiated based on its animal or human host or geographical source, and determine the particular source of the protozoa contamination in source water. The report is not indexed. c. Book News Inc.

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Page Count

140

Publisher

American Water Works Association

Published Date

ISBN 10

1583211241

ISBN 13

9781583211243

Financial and Managerial Accounting

By: Jerry J. Weygandt,Paul D. Kimmel,Donald E. Kieso

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Financial and Managerial Accounting

By: Jerry J. Weygandt,Paul D. Kimmel,Donald E. Kieso

Financial and Managerial Accounting provides students with a clear introduction to fundamental accounting concepts beginning with the building blocks of the accounting cycle and continuing through financial statements. This product is ideal for a two-semester Financial and Managerial Accounting sequence where students spend equal time learning financial and managerial accounting concepts as well as learn the accounting cycle from a corporate perspective.

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Page Count

1330

Publisher

John Wiley & Sons

Published Date

ISBN 10

1119391512

ISBN 13

9781119391517

Financial & Managerial Accounting

By: Jerry J. Weygandt,Paul D. Kimmel,Donald E. Kieso

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Financial & Managerial Accounting

By: Jerry J. Weygandt,Paul D. Kimmel,Donald E. Kieso

Financial and Managerial Accounting, 2nd Edition provides students with a clear introduction to fundamental accounting concepts. The Second Edition helps students get the most out of their accounting course by making practice simple. Both in the print text and online in WileyPLUS with ORION new opportunities for self-guided practice allow students to check their knowledge of accounting concepts, skills, and problem solving techniques and receive personalized feedback at the question, learning objective, and course level. Newly streamlined learning objectives help students use their study time efficiently by creating a clear connections between the reading and video content, and the practice, homework, and assessments questions. Weygandt, Financial and Managerial Accounting is ideal for a two-semester Financial and Managerial Accounting sequence where students spend equal time learning financial and managerial accounting concepts, and learn the accounting cycle from a corporate perspective. This program begins by introducing students to the building blocks of the accounting cycle and builds to financial statements. *WileyPLUS with ORION is sold separately from the text.

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1418

Publisher

John Wiley & Sons

Published Date

ISBN 10

1118334264

ISBN 13

9781118334263

Letter & Spirit, Vol. 11

"Our Beloved Brother Paul" -- Reception History of Paul in Catholic Tradition

By: for Biblical Theology St. Paul Center,Benjamin P. Blosser

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Letter & Spirit, Vol. 11

"Our Beloved Brother Paul" -- Reception History of Paul in Catholic Tradition

By: for Biblical Theology St. Paul Center,Benjamin P. Blosser

The St. Paul Center for Biblical Theology and Dr. Scott Hahn present the eleventh annual edition of Letter & Spirit with the theme "Our Beloved Brother Paul." The articles, while academic in nature, are easily accessible to the average reader and can be read with great profit, both spiritually and in coming to learn the truths of the Catholic faith more deeply.

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222

Published Date

ISBN 10

1945125063

ISBN 13

9781945125065

Book's by Paul A. Rochelle

Comparing Cell Culture and Mouse Assays for Measuring Infectivity of Cryptosporidium

Comparing Cell Culture and Mouse Assays for Measuring Infectivity of Cryptosporidium

Evaluation of Genotyping Techniques for Crytosporidium Parvum

Evaluation of Genotyping Techniques for Crytosporidium Parvum

Financial and Managerial Accounting

Financial and Managerial Accounting

Financial & Managerial Accounting

Financial & Managerial Accounting

Letter & Spirit, Vol. 11

Letter & Spirit, Vol. 11

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