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Impact of confinement housing on study end-points in the calf model of cryptosporidiosis

Identifieur interne : 000698 ( Ncbi/Merge ); précédent : 000697; suivant : 000699

Impact of confinement housing on study end-points in the calf model of cryptosporidiosis

Auteurs : Geneva Graef [États-Unis] ; Natalie J. Hurst [États-Unis] ; Lance Kidder [États-Unis] ; Tracy L. Sy [États-Unis] ; Laura B. Goodman [États-Unis] ; Whitney D. Preston [États-Unis] ; Samuel L. M. Arnold [États-Unis] ; Jennifer A. Zambriski [États-Unis]

Source :

RBID : PMC:5937795

Abstract

Background

Diarrhea is the second leading cause of death in children < 5 years globally and the parasite genus Cryptosporidium is a leading cause of that diarrhea. The global disease burden attributable to cryptosporidiosis is substantial and the only approved chemotherapeutic, nitazoxanide, has poor efficacy in HIV positive children. Chemotherapeutic development is dependent on the calf model of cryptosporidiosis, which is the best approximation of human disease. However, the model is not consistently applied across research studies. Data collection commonly occurs using two different methods: Complete Fecal Collection (CFC), which requires use of confinement housing, and Interval Collection (IC), which permits use of box stalls. CFC mimics human challenge model methodology but it is unknown if confinement housing impacts study end-points and if data gathered via this method is suitable for generalization to human populations.

Methods

Using a modified crossover study design we compared CFC and IC and evaluated the impact of housing on study end-points. At birth, calves were randomly assigned to confinement (n = 14) or box stall housing (n = 9), or were challenged with 5 x 107C. parvum oocysts, and followed for 10 days. Study end-points included fecal oocyst shedding, severity of diarrhea, degree of dehydration, and plasma cortisol.

Findings

Calves in confinement had no significant differences in mean log oocysts enumerated per gram of fecal dry matter between CFC and IC samples (P = 0.6), nor were there diurnal variations in oocyst shedding (P = 0.1). Confinement housed calves shed significantly more oocysts (P = 0.05), had higher plasma cortisol (P = 0.001), and required more supportive care (P = 0.0009) than calves in box stalls.

Conclusion

Housing method confounds study end-points in the calf model of cryptosporidiosis. Due to increased stress data collected from calves in confinement housing may not accurately estimate the efficacy of chemotherapeutics targeting C. parvum.


Url:
DOI: 10.1371/journal.pntd.0006295
PubMed: 29694356
PubMed Central: 5937795

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PMC:5937795

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<title>Background</title>
<p>Diarrhea is the second leading cause of death in children < 5 years globally and the parasite genus
<italic>Cryptosporidium</italic>
is a leading cause of that diarrhea. The global disease burden attributable to cryptosporidiosis is substantial and the only approved chemotherapeutic, nitazoxanide, has poor efficacy in HIV positive children. Chemotherapeutic development is dependent on the calf model of cryptosporidiosis, which is the best approximation of human disease. However, the model is not consistently applied across research studies. Data collection commonly occurs using two different methods: Complete Fecal Collection (CFC), which requires use of confinement housing, and Interval Collection (IC), which permits use of box stalls. CFC mimics human challenge model methodology but it is unknown if confinement housing impacts study end-points and if data gathered via this method is suitable for generalization to human populations.</p>
</sec>
<sec id="sec002">
<title>Methods</title>
<p>Using a modified crossover study design we compared CFC and IC and evaluated the impact of housing on study end-points. At birth, calves were randomly assigned to confinement (n = 14) or box stall housing (n = 9), or were challenged with 5 x 10
<sup>7</sup>
<italic>C</italic>
.
<italic>parvum</italic>
oocysts, and followed for 10 days. Study end-points included fecal oocyst shedding, severity of diarrhea, degree of dehydration, and plasma cortisol.</p>
</sec>
<sec id="sec003">
<title>Findings</title>
<p>Calves in confinement had no significant differences in mean log oocysts enumerated per gram of fecal dry matter between CFC and IC samples (
<italic>P</italic>
= 0.6), nor were there diurnal variations in oocyst shedding (
<italic>P</italic>
= 0.1). Confinement housed calves shed significantly more oocysts (
<italic>P</italic>
= 0.05), had higher plasma cortisol (
<italic>P</italic>
= 0.001), and required more supportive care (
<italic>P</italic>
= 0.0009) than calves in box stalls.</p>
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<p>Housing method confounds study end-points in the calf model of cryptosporidiosis. Due to increased stress data collected from calves in confinement housing may not accurately estimate the efficacy of chemotherapeutics targeting
<italic>C</italic>
.
<italic>parvum</italic>
.</p>
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<journal-id journal-id-type="nlm-ta">PLoS Negl Trop Dis</journal-id>
<journal-id journal-id-type="iso-abbrev">PLoS Negl Trop Dis</journal-id>
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<issn pub-type="ppub">1935-2727</issn>
<issn pub-type="epub">1935-2735</issn>
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<article-id pub-id-type="pmid">29694356</article-id>
<article-id pub-id-type="pmc">5937795</article-id>
<article-id pub-id-type="doi">10.1371/journal.pntd.0006295</article-id>
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</subj-group>
</subj-group>
</subj-group>
</subj-group>
</subj-group>
</subj-group>
<subj-group subj-group-type="Discipline-v3">
<subject>Biology and Life Sciences</subject>
<subj-group>
<subject>Organisms</subject>
<subj-group>
<subject>Eukaryota</subject>
<subj-group>
<subject>Protozoans</subject>
<subj-group>
<subject>Cryptosporidium</subject>
</subj-group>
</subj-group>
</subj-group>
</subj-group>
</subj-group>
<subj-group subj-group-type="Discipline-v3">
<subject>Biology and Life Sciences</subject>
<subj-group>
<subject>Anatomy</subject>
<subj-group>
<subject>Body Fluids</subject>
<subj-group>
<subject>Urine</subject>
</subj-group>
</subj-group>
</subj-group>
</subj-group>
<subj-group subj-group-type="Discipline-v3">
<subject>Medicine and Health Sciences</subject>
<subj-group>
<subject>Anatomy</subject>
<subj-group>
<subject>Body Fluids</subject>
<subj-group>
<subject>Urine</subject>
</subj-group>
</subj-group>
</subj-group>
</subj-group>
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<subject>Biology and Life Sciences</subject>
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<subject>Body Fluids</subject>
<subj-group>
<subject>Urine</subject>
</subj-group>
</subj-group>
</subj-group>
</subj-group>
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<title-group>
<article-title>Impact of confinement housing on study end-points in the calf model of cryptosporidiosis</article-title>
<alt-title alt-title-type="running-head">Impact of confinement housing in the calf model of cryptosporidiosis</alt-title>
</title-group>
<contrib-group>
<contrib contrib-type="author" equal-contrib="yes">
<name>
<surname>Graef</surname>
<given-names>Geneva</given-names>
</name>
<role content-type="http://credit.casrai.org/">Conceptualization</role>
<role content-type="http://credit.casrai.org/">Data curation</role>
<role content-type="http://credit.casrai.org/">Formal analysis</role>
<role content-type="http://credit.casrai.org/">Investigation</role>
<role content-type="http://credit.casrai.org/">Methodology</role>
<role content-type="http://credit.casrai.org/">Project administration</role>
<role content-type="http://credit.casrai.org/">Writing – original draft</role>
<xref ref-type="aff" rid="aff001">
<sup>1</sup>
</xref>
</contrib>
<contrib contrib-type="author">
<name>
<surname>Hurst</surname>
<given-names>Natalie J.</given-names>
</name>
<role content-type="http://credit.casrai.org/">Data curation</role>
<role content-type="http://credit.casrai.org/">Investigation</role>
<role content-type="http://credit.casrai.org/">Methodology</role>
<role content-type="http://credit.casrai.org/">Writing – review & editing</role>
<xref ref-type="aff" rid="aff001">
<sup>1</sup>
</xref>
<xref ref-type="author-notes" rid="econtrib001">
<sup></sup>
</xref>
</contrib>
<contrib contrib-type="author">
<name>
<surname>Kidder</surname>
<given-names>Lance</given-names>
</name>
<role content-type="http://credit.casrai.org/">Data curation</role>
<role content-type="http://credit.casrai.org/">Methodology</role>
<role content-type="http://credit.casrai.org/">Writing – review & editing</role>
<xref ref-type="aff" rid="aff001">
<sup>1</sup>
</xref>
<xref ref-type="author-notes" rid="econtrib001">
<sup></sup>
</xref>
</contrib>
<contrib contrib-type="author">
<name>
<surname>Sy</surname>
<given-names>Tracy L.</given-names>
</name>
<role content-type="http://credit.casrai.org/">Conceptualization</role>
<role content-type="http://credit.casrai.org/">Data curation</role>
<role content-type="http://credit.casrai.org/">Investigation</role>
<role content-type="http://credit.casrai.org/">Methodology</role>
<role content-type="http://credit.casrai.org/">Project administration</role>
<role content-type="http://credit.casrai.org/">Supervision</role>
<role content-type="http://credit.casrai.org/">Writing – review & editing</role>
<xref ref-type="aff" rid="aff001">
<sup>1</sup>
</xref>
<xref ref-type="author-notes" rid="currentaff001">
<sup>¤</sup>
</xref>
<xref ref-type="author-notes" rid="econtrib001">
<sup></sup>
</xref>
</contrib>
<contrib contrib-type="author">
<name>
<surname>Goodman</surname>
<given-names>Laura B.</given-names>
</name>
<role content-type="http://credit.casrai.org/">Formal analysis</role>
<role content-type="http://credit.casrai.org/">Methodology</role>
<role content-type="http://credit.casrai.org/">Validation</role>
<role content-type="http://credit.casrai.org/">Writing – review & editing</role>
<xref ref-type="aff" rid="aff002">
<sup>2</sup>
</xref>
</contrib>
<contrib contrib-type="author">
<name>
<surname>Preston</surname>
<given-names>Whitney D.</given-names>
</name>
<role content-type="http://credit.casrai.org/">Data curation</role>
<role content-type="http://credit.casrai.org/">Project administration</role>
<role content-type="http://credit.casrai.org/">Supervision</role>
<role content-type="http://credit.casrai.org/">Writing – review & editing</role>
<xref ref-type="aff" rid="aff001">
<sup>1</sup>
</xref>
</contrib>
<contrib contrib-type="author">
<name>
<surname>Arnold</surname>
<given-names>Samuel L. M.</given-names>
</name>
<role content-type="http://credit.casrai.org/">Data curation</role>
<role content-type="http://credit.casrai.org/">Validation</role>
<role content-type="http://credit.casrai.org/">Writing – review & editing</role>
<xref ref-type="aff" rid="aff003">
<sup>3</sup>
</xref>
</contrib>
<contrib contrib-type="author" equal-contrib="yes">
<contrib-id authenticated="true" contrib-id-type="orcid">http://orcid.org/0000-0003-0948-1181</contrib-id>
<name>
<surname>Zambriski</surname>
<given-names>Jennifer A.</given-names>
</name>
<role content-type="http://credit.casrai.org/">Conceptualization</role>
<role content-type="http://credit.casrai.org/">Data curation</role>
<role content-type="http://credit.casrai.org/">Formal analysis</role>
<role content-type="http://credit.casrai.org/">Funding acquisition</role>
<role content-type="http://credit.casrai.org/">Investigation</role>
<role content-type="http://credit.casrai.org/">Methodology</role>
<role content-type="http://credit.casrai.org/">Project administration</role>
<role content-type="http://credit.casrai.org/">Supervision</role>
<role content-type="http://credit.casrai.org/">Writing – original draft</role>
<xref ref-type="aff" rid="aff001">
<sup>1</sup>
</xref>
<xref ref-type="corresp" rid="cor001">*</xref>
</contrib>
</contrib-group>
<aff id="aff001">
<label>1</label>
<addr-line>Paul G. Allen School for Global Animal Health, College of Veterinary Medicine, Washington State University, Pullman, Washington, United States of America</addr-line>
</aff>
<aff id="aff002">
<label>2</label>
<addr-line>Department of Population Medicine and Diagnostic Sciences, College of Veterinary Medicine, Cornell University, Ithaca, New York, United States of America</addr-line>
</aff>
<aff id="aff003">
<label>3</label>
<addr-line>Department of Allergy and Infectious Disease, School of Medicine, University of Washington, Seattle, Washington, United States of America</addr-line>
</aff>
<contrib-group>
<contrib contrib-type="editor">
<name>
<surname>Pavlinac</surname>
<given-names>Patricia</given-names>
</name>
<role>Editor</role>
<xref ref-type="aff" rid="edit1"></xref>
</contrib>
</contrib-group>
<aff id="edit1">
<addr-line>University of Washington, UNITED STATES</addr-line>
</aff>
<author-notes>
<fn fn-type="COI-statement" id="coi001">
<p>The authors have declared that no competing interests exist.</p>
</fn>
<fn fn-type="current-aff" id="currentaff001">
<label>¤</label>
<p>Current address: College of Veterinary Medicine, Colorado State University, Ft. Collins, Colorado, United States of America</p>
</fn>
<fn fn-type="other" id="econtrib001">
<p>‡ NJH, LK, and TLS also contributed equally to this work.</p>
</fn>
<corresp id="cor001">* E-mail:
<email>jzambriski@vetmed.wsu.edu</email>
</corresp>
</author-notes>
<pub-date pub-type="epub">
<day>25</day>
<month>4</month>
<year>2018</year>
</pub-date>
<pub-date pub-type="collection">
<month>4</month>
<year>2018</year>
</pub-date>
<volume>12</volume>
<issue>4</issue>
<elocation-id>e0006295</elocation-id>
<history>
<date date-type="received">
<day>13</day>
<month>6</month>
<year>2017</year>
</date>
<date date-type="accepted">
<day>2</day>
<month>2</month>
<year>2018</year>
</date>
</history>
<permissions>
<copyright-statement>© 2018 Graef et al</copyright-statement>
<copyright-year>2018</copyright-year>
<copyright-holder>Graef et al</copyright-holder>
<license xlink:href="http://creativecommons.org/licenses/by/4.0/">
<license-p>This is an open access article distributed under the terms of the
<ext-link ext-link-type="uri" xlink:href="http://creativecommons.org/licenses/by/4.0/">Creative Commons Attribution License</ext-link>
, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.</license-p>
</license>
</permissions>
<self-uri content-type="pdf" xlink:href="pntd.0006295.pdf"></self-uri>
<abstract>
<sec id="sec001">
<title>Background</title>
<p>Diarrhea is the second leading cause of death in children < 5 years globally and the parasite genus
<italic>Cryptosporidium</italic>
is a leading cause of that diarrhea. The global disease burden attributable to cryptosporidiosis is substantial and the only approved chemotherapeutic, nitazoxanide, has poor efficacy in HIV positive children. Chemotherapeutic development is dependent on the calf model of cryptosporidiosis, which is the best approximation of human disease. However, the model is not consistently applied across research studies. Data collection commonly occurs using two different methods: Complete Fecal Collection (CFC), which requires use of confinement housing, and Interval Collection (IC), which permits use of box stalls. CFC mimics human challenge model methodology but it is unknown if confinement housing impacts study end-points and if data gathered via this method is suitable for generalization to human populations.</p>
</sec>
<sec id="sec002">
<title>Methods</title>
<p>Using a modified crossover study design we compared CFC and IC and evaluated the impact of housing on study end-points. At birth, calves were randomly assigned to confinement (n = 14) or box stall housing (n = 9), or were challenged with 5 x 10
<sup>7</sup>
<italic>C</italic>
.
<italic>parvum</italic>
oocysts, and followed for 10 days. Study end-points included fecal oocyst shedding, severity of diarrhea, degree of dehydration, and plasma cortisol.</p>
</sec>
<sec id="sec003">
<title>Findings</title>
<p>Calves in confinement had no significant differences in mean log oocysts enumerated per gram of fecal dry matter between CFC and IC samples (
<italic>P</italic>
= 0.6), nor were there diurnal variations in oocyst shedding (
<italic>P</italic>
= 0.1). Confinement housed calves shed significantly more oocysts (
<italic>P</italic>
= 0.05), had higher plasma cortisol (
<italic>P</italic>
= 0.001), and required more supportive care (
<italic>P</italic>
= 0.0009) than calves in box stalls.</p>
</sec>
<sec id="sec004">
<title>Conclusion</title>
<p>Housing method confounds study end-points in the calf model of cryptosporidiosis. Due to increased stress data collected from calves in confinement housing may not accurately estimate the efficacy of chemotherapeutics targeting
<italic>C</italic>
.
<italic>parvum</italic>
.</p>
</sec>
</abstract>
<abstract abstract-type="summary">
<title>Author summary</title>
<p>After respiratory disease, diarrhea is the leading cause of death in children < 5 years worldwide. Recent multicenter studies have identified the parasite genus,
<italic>Cryptosporidium</italic>
to be one of the most common causes of pediatric diarrhea in resource-poor settings. In industrialized nations the parasite is well controlled through centralized water treatment facilities. Consequently, drug development targeting this neglected tropical disease has been limited. There are currently no vaccines to prevent cryptosporidiosis. Nitazoxanide is the only approved drug targeting
<italic>Cryptosporidium</italic>
, but it has poor efficacy in HIV positive children. Owing to the recently published multicenter findings there have been renewed
<italic>Cryptosporidium</italic>
drug development efforts. The calf model of cryptosporidiosis is widely recognized as the best approximation of human disease but it is difficult to execute resulting in varied experimental approaches. Here, we describe the two most common methods of executing the calf model of cryptosporidiosis and investigate how these methods impact study end-points used to inform identification of drug candidates and clinical trials. We find that confinement housing significantly increases plasma cortisol in calves and impacts key study end-points. Data collected via confinement housing may not accurately estimate drug efficacy, underscoring the need for model harmonization. The results of this study should be considered when interpreting chemotherapeutic data derived from the calf model of cryptosporidiosis.</p>
</abstract>
<funding-group>
<award-group id="award001">
<funding-source>
<institution-wrap>
<institution-id institution-id-type="funder-id">http://dx.doi.org/10.13039/100007588</institution-id>
<institution>Washington State University</institution>
</institution-wrap>
</funding-source>
<principal-award-recipient>
<contrib-id authenticated="true" contrib-id-type="orcid">http://orcid.org/0000-0003-0948-1181</contrib-id>
<name>
<surname>Zambriski</surname>
<given-names>Jennifer A.</given-names>
</name>
</principal-award-recipient>
</award-group>
<funding-statement>This research was supported by institutional start-up funds. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.</funding-statement>
</funding-group>
<counts>
<fig-count count="4"></fig-count>
<table-count count="6"></table-count>
<page-count count="20"></page-count>
</counts>
<custom-meta-group>
<custom-meta>
<meta-name>PLOS Publication Stage</meta-name>
<meta-value>vor-update-to-uncorrected-proof</meta-value>
</custom-meta>
<custom-meta>
<meta-name>Publication Update</meta-name>
<meta-value>2018-05-07</meta-value>
</custom-meta>
<custom-meta id="data-availability">
<meta-name>Data Availability</meta-name>
<meta-value>Data are available from the Dryad Digital Repository (
<ext-link ext-link-type="uri" xlink:href="https://datadryad.org">https://datadryad.org</ext-link>
) (doi:
<ext-link ext-link-type="uri" xlink:href="https://doi.org/10.5061/dryad.hg2g312">10.5061/dryad.hg2g312</ext-link>
).</meta-value>
</custom-meta>
</custom-meta-group>
</article-meta>
<notes>
<title>Data Availability</title>
<p>Data are available from the Dryad Digital Repository (
<ext-link ext-link-type="uri" xlink:href="https://datadryad.org">https://datadryad.org</ext-link>
) (doi:
<ext-link ext-link-type="uri" xlink:href="https://doi.org/10.5061/dryad.hg2g312">10.5061/dryad.hg2g312</ext-link>
).</p>
</notes>
</front>
</pmc>
<affiliations>
<list>
<country>
<li>États-Unis</li>
</country>
<region>
<li>Washington (État)</li>
<li>État de New York</li>
</region>
<settlement>
<li>Ithaca (New York)</li>
<li>Seattle</li>
</settlement>
<orgName>
<li>Université Cornell</li>
<li>Université de Washington</li>
</orgName>
</list>
<tree>
<country name="États-Unis">
<region name="Washington (État)">
<name sortKey="Graef, Geneva" sort="Graef, Geneva" uniqKey="Graef G" first="Geneva" last="Graef">Geneva Graef</name>
</region>
<name sortKey="Arnold, Samuel L M" sort="Arnold, Samuel L M" uniqKey="Arnold S" first="Samuel L. M." last="Arnold">Samuel L. M. Arnold</name>
<name sortKey="Goodman, Laura B" sort="Goodman, Laura B" uniqKey="Goodman L" first="Laura B." last="Goodman">Laura B. Goodman</name>
<name sortKey="Hurst, Natalie J" sort="Hurst, Natalie J" uniqKey="Hurst N" first="Natalie J." last="Hurst">Natalie J. Hurst</name>
<name sortKey="Kidder, Lance" sort="Kidder, Lance" uniqKey="Kidder L" first="Lance" last="Kidder">Lance Kidder</name>
<name sortKey="Preston, Whitney D" sort="Preston, Whitney D" uniqKey="Preston W" first="Whitney D." last="Preston">Whitney D. Preston</name>
<name sortKey="Sy, Tracy L" sort="Sy, Tracy L" uniqKey="Sy T" first="Tracy L." last="Sy">Tracy L. Sy</name>
<name sortKey="Zambriski, Jennifer A" sort="Zambriski, Jennifer A" uniqKey="Zambriski J" first="Jennifer A." last="Zambriski">Jennifer A. Zambriski</name>
</country>
</tree>
</affiliations>
</record>

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