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Author: Georgia Wilke
Publisher:
ISBN:
Category : Electronic dissertations
Languages : en
Pages : 209
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Book Description
Cryptosporidium is a genus of protozoan parasites that causes diarrheal disease in humans and other animals. There are two major species that cause disease in humans: C. parvum, which infects both humans and animals, and C. hominis, which primarily infects humans. A recent study investigating the etiologies of pediatric diarrheal illness in Africa and South Asia found that Cryptosporidium is the 2nd most prevalent cause of diarrhea in infants and may be a contributing factor to chronic malnutrition. This discovery has led to renewed interest in studying this parasite and a reexamination of the barriers to studying Cryptosporidium. The main obstacle hindering research on this parasite is that it cannot be propagated in vitro and instead must be passaged through large animals such as calves to generate infectious oocysts. The cell culture models that are available rely on adenocarcinoma cells and only support a few days of growth and do not enable complete life cycle development in vitro. These limitations have stalled the development of research tools for investigating Cryptosporidium biology and have also slowed developmental progress of new therapies.
Author: Georgia Wilke
Publisher:
ISBN:
Category : Electronic dissertations
Languages : en
Pages : 209
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Book Description
Cryptosporidium is a genus of protozoan parasites that causes diarrheal disease in humans and other animals. There are two major species that cause disease in humans: C. parvum, which infects both humans and animals, and C. hominis, which primarily infects humans. A recent study investigating the etiologies of pediatric diarrheal illness in Africa and South Asia found that Cryptosporidium is the 2nd most prevalent cause of diarrhea in infants and may be a contributing factor to chronic malnutrition. This discovery has led to renewed interest in studying this parasite and a reexamination of the barriers to studying Cryptosporidium. The main obstacle hindering research on this parasite is that it cannot be propagated in vitro and instead must be passaged through large animals such as calves to generate infectious oocysts. The cell culture models that are available rely on adenocarcinoma cells and only support a few days of growth and do not enable complete life cycle development in vitro. These limitations have stalled the development of research tools for investigating Cryptosporidium biology and have also slowed developmental progress of new therapies.
Author: G. Widmer
Publisher: IWA Publishing
ISBN: 9781843399483
Category : Science
Languages : en
Pages : 62
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Book Description
The goal of this project was to investigate methods, which promote growth of Cryptosporidium (C.) parvum in culture. The researchers tested the assumption that improved survival of host cells enhances growth of C. parvum. Although certain modifications to the culture methodology improved host cell survival, enhancing host cell survival was not always accompanied by an increase in parasite density in the cell monolayer. Some cultures grown on collagen or laminin showed improved parasite growth. However, this effect was not consistent, indicating that other variables affect parasite growth. C. parvum was more prevalent in cells in the mitotic cycle than in non-dividing cells. This is consistent with the absence of certain biosynthetic pathways in C. parvum, and suggests that dividing cells may be more favorable to parasite development. Although current culture methods enable the detection of infectious Cryptosporidium oocysts, limited parasite growth reduces the sensitivity of the assay, making it necessary to apply sophisticated methods for the detection of the intracellular parasites. This study identified methods that slightly improve growth of Cryptosporidium in culture and also highlighted methods that have no effect. This report is available only as a pay-per-view item.
Author: Paul A. Rochelle
Publisher: IWA Publishing
ISBN: 1843399156
Category : Science
Languages : en
Pages : 136
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Book Description
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
Author:
Publisher:
ISBN:
Category : Cryptosporidium parvum
Languages : en
Pages : 45
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Book Description
Author: Guadalupe Ortega-Pierres
Publisher: CABI
ISBN: 1845933915
Category : Medical
Languages : en
Pages : 518
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Book Description
Giardia and Cryptosporidium are both parasites of considerable global interest due to the gastrointestinal problems these organisms can cause in humans as well as domestic and wild animals. Despite its long history as the causative agent of giardiasis, the clinical and zoonotic significance of Giardia infections in humans and animals is only beginning to emerge with the application of molecular tools. Less is known about Cryptosporidium and studies seek to understand the impact of infection in livestock and other animals, its epidemiology and zoonotic significance. Presenting a comprehensive overview of recent research this book draws on the experience of experts in all fields of Giardia and Cryptosporidium research.
Author: Jan R. Mead
Publisher: Humana
ISBN: 9781493997473
Category : Medical
Languages : en
Pages : 407
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Book Description
This book encompasses broad aspects of Cryptosporidium research with established methods that have been improved and expanded over the years as well as recent cutting-edge techniques. Within this collection are numerous molecular methods as well as protocols for genotyping and diagnostics, while also including room for in vitro cell culture techniques to address the issues with growing this difficult organism continuously. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Cryptosporidium: Methods and Protocols serves as an idea guide to the inherent challenges of working with cryptosporidial parasites to provide a foundation for new investigators to build upon. The chapter “Accessing Cryptosporidium Omic and Isolate Data via CryptoDB.org” is available open access under a Creative Commons Attribution 4.0 International License via link.springer.com.
Author: Ronald Fayer
Publisher: CRC Press
ISBN: 1420052276
Category : Medical
Languages : en
Pages : 579
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Book Description
From the microscopic observation of infection to the widespread application of molecular techniques in taxonomy and epidemiology, to the genome sequencing of two major species and advances in biochemistry, phylogeny, and water treatment, new information on this fascinating genus continues to mount as we discover and utilize the latest scientific te
Author: E. Scholtyseck
Publisher: Springer Science & Business Media
ISBN: 3642670679
Category : Science
Languages : en
Pages : 212
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Book Description
The plan for this atlas evolved from the necessity of providing the biology student interested in protozoology, cytology, and para sitology with an introduction to the study of fine structure in Protozoa. To reduce the book's extend, a selection of characteristic protozoans had to be made, limited to those which could be regarded as representative for entire groups. Interest in parasitic protozoans has been steadily on the increase over the last 10 years. This particular group of organisms thus seemed a very suitable choice. The" Apicomplexa" were selected as an area of emphasis. These once were part of the collective group called Sporozoa which included many parasitic protozoans of uncertain taxonomy. Fine structural research has been of especial significance for the Apicomplexa, since Protozoa belonging to this subgroup can now be named, characterized, and classified by features recognizable by electron microscopy. Only the fine structure of whole cells is represented in this atlas, so that the ciliates have been of necessity excluded. Their cells are too large in diameter in any case for our purpose here. They also play only a minor role as parasitic organisms. This book utilizes a new method to facilitate the analysis of protozoan fine structure. An electron micrograph, a descriptive text, and an analytic drawing are arranged on two facing pages so that the electron micrograph and the drawing can be compared.
Author: Franz Petry
Publisher: Karger Medical and Scientific Publishers
ISBN: 3805570503
Category : Medical
Languages : en
Pages : 279
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Book Description
'... This volume provides most complete and balanced coverage of essential aspects of the pathogens as well as the diagnosis and clinical correlations of the disease they cause. It is set to become a valuable refrence for parasitologists, protozoologists, molecular biologists, clinical mecrobiologists, epidemiologists and specialists in infectious diseases.'
Author: R.C.A. Thompson
Publisher: Elsevier
ISBN: 0080530109
Category : Science
Languages : en
Pages : 469
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Book Description
In the relatively short period since Cryptosporidium was recognised as a human pathogen, and that it could be transmitted in water as well as directly between animals and people, it has been the subject of intense investigations. Its status as an opportunistic pathogen, especially in AIDS patients, and the lack of effective anti-cryptosporidial drugs have served to emphasise the public health importance of this organism. This has to some extent overshadowed the fact that Cryptosporidium is also an important pathogen of domestic animals and wildlife. In recent years, the application of molecular biology and culture techniques have had an enormous impact on our understanding of the aetiological agents of cryptosporidial infections and our ability to study the causative agents in the laboratory. As a consequence, a wealth of information and novel data has been produced during the last 3-4 years, particularly in the areas of taxonomy, biology, pathogenesis, epidemiology - particularly zoonotic and water borne transmission, and treatment. It is thus very timely to bring together in this book the international research community involved to review the major advances in research and identify the important research priorities for the future, thus enabling as wide an audience as possible to benefit from and share in this comprehensive look at Cryptosporidium and cryptosporidiosis.
