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<front>
<journal-meta>
<journal-id journal-id-type="publisher-id">Front. Vet. Sci.</journal-id>
<journal-title-group>
<journal-title>Frontiers in Veterinary Science</journal-title>
<abbrev-journal-title abbrev-type="pubmed">Front. Vet. Sci.</abbrev-journal-title>
</journal-title-group>
<issn pub-type="epub">2297-1769</issn>
<publisher>
<publisher-name>Frontiers Media S.A.</publisher-name>
</publisher>
</journal-meta>
<article-meta>
<article-id pub-id-type="doi">10.3389/fvets.2025.1732237</article-id>
<article-version article-version-type="Version of Record" vocab="NISO-RP-8-2008"/>
<article-categories>
<subj-group subj-group-type="heading">
<subject>Original Research</subject>
</subj-group>
</article-categories>
<title-group>
<article-title>Tear film characteristics in French bulldogs vs. non-brachycephalic dogs</article-title>
</title-group>
<contrib-group>
<contrib contrib-type="author" corresp="yes">
<name>
<surname>Spornberger</surname>
<given-names>Joschka</given-names>
</name>
<xref ref-type="aff" rid="aff1"><sup>1</sup></xref>
<xref ref-type="corresp" rid="c001"><sup>&#x002A;</sup></xref>
<uri xlink:href="https://loop.frontiersin.org/people/3234157"/>
<role vocab="credit" vocab-identifier="https://credit.niso.org/" vocab-term="conceptualization" vocab-term-identifier="https://credit.niso.org/contributor-roles/conceptualization/">Conceptualization</role>
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</contrib>
<contrib contrib-type="author">
<name>
<surname>Soukup</surname>
<given-names>Petr</given-names>
</name>
<xref ref-type="aff" rid="aff1"><sup>1</sup></xref>
<uri xlink:href="https://loop.frontiersin.org/people/3234808"/>
<role vocab="credit" vocab-identifier="https://credit.niso.org/" vocab-term="Formal analysis" vocab-term-identifier="https://credit.niso.org/contributor-roles/formal-analysis/">Formal analysis</role>
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</contrib>
<contrib contrib-type="author">
<name>
<surname>Meyer-Lindenberg</surname>
<given-names>Andrea</given-names>
</name>
<xref ref-type="aff" rid="aff2"><sup>2</sup></xref>
<uri xlink:href="https://loop.frontiersin.org/people/515413"/>
<role vocab="credit" vocab-identifier="https://credit.niso.org/" vocab-term="methodology" vocab-term-identifier="https://credit.niso.org/contributor-roles/methodology/">Methodology</role>
<role vocab="credit" vocab-identifier="https://credit.niso.org/" vocab-term="supervision" vocab-term-identifier="https://credit.niso.org/contributor-roles/supervision/">Supervision</role>
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</contrib>
<contrib contrib-type="author">
<name>
<surname>Allgoewer</surname>
<given-names>Ingrid</given-names>
</name>
<xref ref-type="aff" rid="aff1"><sup>1</sup></xref>
<uri xlink:href="https://loop.frontiersin.org/people/3284851"/>
<role vocab="credit" vocab-identifier="https://credit.niso.org/" vocab-term="conceptualization" vocab-term-identifier="https://credit.niso.org/contributor-roles/conceptualization/">Conceptualization</role>
<role vocab="credit" vocab-identifier="https://credit.niso.org/" vocab-term="Formal analysis" vocab-term-identifier="https://credit.niso.org/contributor-roles/formal-analysis/">Formal analysis</role>
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<aff id="aff1"><label>1</label><institution>Animal Eye Practice</institution>, <city>Berlin</city>, <country country="de">Germany</country></aff>
<aff id="aff2"><label>2</label><institution>Clinic for Small Animal Surgery and Reproduction, Centre for Clinical Veterinary Medicine, LMU Munich</institution>, <city>Munich</city>, <country country="de">Germany</country></aff>
<author-notes>
<corresp id="c001"><label>&#x002A;</label>Correspondence: Joschka Spornberger, <email xlink:href="mailto:J.Spornberger@gmail.com">J.Spornberger@gmail.com</email></corresp>
</author-notes>
<pub-date publication-format="electronic" date-type="pub" iso-8601-date="2025-12-10">
<day>10</day>
<month>12</month>
<year>2025</year>
</pub-date>
<pub-date publication-format="electronic" date-type="collection">
<year>2025</year>
</pub-date>
<volume>12</volume>
<elocation-id>1732237</elocation-id>
<history>
<date date-type="received">
<day>25</day>
<month>10</month>
<year>2025</year>
</date>
<date date-type="rev-recd">
<day>22</day>
<month>11</month>
<year>2025</year>
</date>
<date date-type="accepted">
<day>25</day>
<month>11</month>
<year>2025</year>
</date>
</history>
<permissions>
<copyright-statement>Copyright &#x00A9; 2025 Spornberger, Soukup, Meyer-Lindenberg and Allgoewer.</copyright-statement>
<copyright-year>2025</copyright-year>
<copyright-holder>Spornberger, Soukup, Meyer-Lindenberg and Allgoewer</copyright-holder>
<license>
<ali:license_ref start_date="2025-12-10">https://creativecommons.org/licenses/by/4.0/</ali:license_ref>
<license-p>This is an open-access article distributed under the terms of the <ext-link ext-link-type="uri" xlink:href="https://creativecommons.org/licenses/by/4.0/">Creative Commons Attribution License (CC BY)</ext-link>. The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.</license-p>
</license>
</permissions>
<abstract>
<sec>
<title>Purpose</title>
<p>To investigate the tear film characteristics of healthy French Bulldogs and compare them with healthy dogs of mesocephalic and dolichocephalic breeds.</p>
</sec>
<sec>
<title>Methods</title>
<p>French Bulldogs and non-brachycephalic dogs considered healthy underwent a complete ophthalmic examination. Tear osmolarity was tested with the ScoutPro&#x00AE; osmolarity system and a tear film analysis, including interferometry, non-invasive tear film break-up time (NIBUT), tear meniscus height measurement (TMH) and meibography using the I.C.P. OSA-Vet&#x00AE; was performed.</p>
</sec>
<sec>
<title>Results</title>
<p>French Bulldogs had a significantly higher Schirmer Tear Test I (19.05&#x202F;&#x00B1;&#x202F;4.00 vs. 16.88&#x202F;&#x00B1;&#x202F;3.54, <italic>p</italic>&#x202F;=&#x202F;0.017), a higher TMH (0.64&#x202F;&#x00B1;&#x202F;0.20 vs. 0.31&#x202F;&#x00B1;&#x202F;0.22, <italic>p</italic>&#x202F;=&#x202F;9&#x00D7;10<sup>&#x2212;10</sup>), a lower interferometry grading and a reduced NIBUT (7.39&#x202F;&#x00B1;&#x202F;3.32 vs. 14.74&#x202F;&#x00B1;&#x202F;5.92, <italic>p</italic>&#x202F;=&#x202F;3.5&#x00D7;10<sup>&#x2212;9</sup>) compared to the control group. Differences in tear osmolarity were not statistically significant between the groups (296.6&#x202F;&#x00B1;&#x202F;13.6 vs. 299.8&#x202F;&#x00B1;&#x202F;16.1, <italic>p</italic>&#x202F;=&#x202F;0.40).</p>
</sec>
<sec>
<title>Conclusion</title>
<p>The tear film in healthy French Bulldogs shows signs of a qualitative tear film deficiency/evaporative dry eye disease (EDED) compared to non-brachycephalic dogs. The influence of the altered tear film on diseases of the ocular surface in French Bulldogs needs to be further investigated.</p>
</sec>
</abstract>
<kwd-group>
<kwd>brachycephalic dogs</kwd>
<kwd>evaporative dry eye disease</kwd>
<kwd>ocular surface health</kwd>
<kwd>tearfilm</kwd>
<kwd>tear film diagnostics</kwd>
</kwd-group>
<funding-group>
<funding-statement>The author(s) declare that financial support was received for the research and/or publication of this article. The study was funded by the DOK Dortmunder Kreis, Germany.</funding-statement>
</funding-group>
<counts>
<fig-count count="4"/>
<table-count count="2"/>
<equation-count count="0"/>
<ref-count count="60"/>
<page-count count="11"/>
<word-count count="7805"/>
</counts>
<custom-meta-group>
<custom-meta>
<meta-name>section-at-acceptance</meta-name>
<meta-value>Comparative and Clinical Medicine</meta-value>
</custom-meta>
</custom-meta-group>
</article-meta>
</front>
<body>
<sec sec-type="intro" id="sec1">
<label>1</label>
<title>Introduction</title>
<p>Brachycephalic dogs are disproportionately affected by ocular surface diseases (<xref ref-type="bibr" rid="ref1 ref2 ref3 ref4 ref5">1&#x2013;5</xref>). D. G. O&#x2019;Neill et al. showed that brachycephalic dogs have a 3.63 times higher risk of developing keratoconjunctivitis sicca (KCS) than mesocephalic breeds (<xref ref-type="bibr" rid="ref4">4</xref>). This, along with their overrepresentation in veterinary ophthalmology (<xref ref-type="bibr" rid="ref6">6</xref>), is due to anatomic and physiologic changes that are collectively referred to as brachycephalic ocular syndrome (BOS) (<xref ref-type="bibr" rid="ref1">1</xref>, <xref ref-type="bibr" rid="ref3">3</xref>, <xref ref-type="bibr" rid="ref7">7</xref>). However, there is discrepancy regarding the most common ocular surface disease in the various brachycephalic breeds (<xref ref-type="bibr" rid="ref5 ref6 ref7 ref8 ref9 ref10 ref11 ref12 ref13">5&#x2013;13</xref>) (<xref ref-type="table" rid="tab1">Table 1</xref>).</p>
<table-wrap position="float" id="tab1">
<label>Table 1</label>
<caption>
<p>Breed specific prevalence of ocular surface disease in brachycephalic dogs: summary of published studies.</p>
</caption>
<table frame="hsides" rules="groups">
<thead>
<tr>
<th align="left" valign="top">Breed</th>
<th align="left" valign="top">Number of animals/eyes</th>
<th align="left" valign="top">Health status</th>
<th align="left" valign="top">Most prevalent ocular surface disease</th>
<th align="left" valign="top">Study</th>
</tr>
</thead>
<tbody>
<tr>
<td align="left" valign="top" rowspan="3">Shih Tzu</td>
<td align="left" valign="top" rowspan="3">1,012 eyes</td>
<td align="left" valign="top" rowspan="3">ophthalmic disorders</td>
<td align="left" valign="top">KCS (12.5%)</td>
<td align="left" valign="top" rowspan="15">Palmer et al. (<xref ref-type="bibr" rid="ref6">6</xref>)</td>
</tr>
<tr>
<td align="left" valign="top">Ulcer (11.1%)</td>
</tr>
<tr>
<td align="left" valign="top">Corneal pigmentation (10.1%)</td>
</tr>
<tr>
<td align="left" valign="top" rowspan="2">Pug</td>
<td align="left" valign="top" rowspan="2">558 eyes</td>
<td align="left" valign="top" rowspan="2">ophthalmic disorders</td>
<td align="left" valign="top">Corneal pigmentation (22.9%)</td>
</tr>
<tr>
<td align="left" valign="top">KCS (9.1%)</td>
</tr>
<tr>
<td align="left" valign="top">Boston Terrier</td>
<td align="left" valign="top">567 eyes</td>
<td align="left" valign="top">ophthalmic disorders</td>
<td align="left" valign="top">Ulcer (14.8%)</td>
</tr>
<tr>
<td align="left" valign="top">English Bulldog</td>
<td align="left" valign="top">320 eyes</td>
<td align="left" valign="top">ophthalmic disorders</td>
<td align="left" valign="top">Ulcer (13.4%)</td>
</tr>
<tr>
<td align="left" valign="top" rowspan="3">Lhasa Apso</td>
<td align="left" valign="top" rowspan="3">201 eyes</td>
<td align="left" valign="top" rowspan="3">ophthalmic disorders</td>
<td align="left" valign="top">KCS (16.9%)</td>
</tr>
<tr>
<td align="left" valign="top">Ulcer (11.9%)</td>
</tr>
<tr>
<td align="left" valign="top">Corneal pigmentation (7.5%)</td>
</tr>
<tr>
<td align="left" valign="top" rowspan="3">Pekingese</td>
<td align="left" valign="top" rowspan="3">131 eyes</td>
<td align="left" valign="top" rowspan="3">ophthalmic disorders</td>
<td align="left" valign="top">Corneal Pigmentation (17.6%)</td>
</tr>
<tr>
<td align="left" valign="top">Ulcer (17.6%)</td>
</tr>
<tr>
<td align="left" valign="top">KCS (15.3%)</td>
</tr>
<tr>
<td align="left" valign="top" rowspan="2">French Bulldog</td>
<td align="left" valign="top" rowspan="2">84 eyes</td>
<td align="left" valign="top" rowspan="2">ophthalmic disorders</td>
<td align="left" valign="top">Ulcer (20.2%)</td>
</tr>
<tr>
<td align="left" valign="top">KCS (4.8%)</td>
</tr>
<tr>
<td align="left" valign="top">Pug</td>
<td align="left" valign="top">17 animals</td>
<td align="left" valign="top">ophthalmic disorders</td>
<td align="left" valign="top">Corneal pigmentation (53%)</td>
<td align="left" valign="top" rowspan="4">Costa et al. (<xref ref-type="bibr" rid="ref7">7</xref>)</td>
</tr>
<tr>
<td align="left" valign="top">Shih Tzu</td>
<td align="left" valign="top">22 animals</td>
<td align="left" valign="top">ophthalmic disorders</td>
<td align="left" valign="top">Corneal pigmentation (36%)</td>
</tr>
<tr>
<td align="left" valign="top">Shih Tzu</td>
<td align="left" valign="top">22 animals</td>
<td align="left" valign="top">ophthalmic disorders</td>
<td align="left" valign="top">Fibrosis (36%)</td>
</tr>
<tr>
<td align="left" valign="top">French Bulldog</td>
<td align="left" valign="top">38 animals</td>
<td align="left" valign="top">ophthalmic disorders</td>
<td align="left" valign="top">Fibrosis (29%)</td>
</tr>
<tr>
<td align="left" valign="top" rowspan="3">Shih Tzu</td>
<td align="left" valign="top" rowspan="3">1,000 eyes</td>
<td align="left" valign="top" rowspan="3">ophthalmic disorders</td>
<td align="left" valign="top">reduced tear film breakup time (17.6%)</td>
<td align="left" valign="top" rowspan="3">Rajaei et al. (<xref ref-type="bibr" rid="ref8">8</xref>)</td>
</tr>
<tr>
<td align="left" valign="top">KCS (7.8%)</td>
</tr>
<tr>
<td align="left" valign="top">Corneal Pigmentation (7.8%)</td>
</tr>
<tr>
<td align="left" valign="top" rowspan="3">Shih Tzu</td>
<td align="left" valign="top" rowspan="3">50 animals</td>
<td align="left" valign="top" rowspan="3">healthy</td>
<td align="left" valign="top">Corneal Pigmentation (27%)</td>
<td align="left" valign="top" rowspan="3">Sebbag et al. (<xref ref-type="bibr" rid="ref9">9</xref>)</td>
</tr>
<tr>
<td align="left" valign="top">Fibrosis (20%)</td>
</tr>
<tr>
<td align="left" valign="top">reduced tear film breakup time</td>
</tr>
<tr>
<td align="left" valign="top" rowspan="2">Shih Tzu</td>
<td align="left" valign="top" rowspan="2">11 animals</td>
<td align="left" valign="top" rowspan="2">KCS</td>
<td align="left" valign="top">Ulcer (36%)</td>
<td align="left" valign="top" rowspan="4">Sanchez RF. et al. (<xref ref-type="bibr" rid="ref13">13</xref>)</td>
</tr>
<tr>
<td align="left" valign="top">Fibrosis (72%)</td>
</tr>
<tr>
<td align="left" valign="top" rowspan="2">Cavalier King Charles Spaniel</td>
<td align="left" valign="top" rowspan="2">40 animals</td>
<td align="left" valign="top" rowspan="2">KCS</td>
<td align="left" valign="top">Ulcer (30%)</td>
</tr>
<tr>
<td align="left" valign="top">Fibrosis (52.5%)</td>
</tr>
<tr>
<td align="left" valign="top">Pug</td>
<td align="left" valign="top">295</td>
<td align="left" valign="top">ophthalmic disorders</td>
<td align="left" valign="top">Corneal pigmentation (82.4%)</td>
<td/>
</tr>
<tr>
<td align="left" valign="top">Pug</td>
<td align="left" valign="top">1,015 animals</td>
<td align="left" valign="top">ophthalmic and general disorders</td>
<td align="left" valign="top">Ulcer (5.42%)</td>
<td align="left" valign="top" rowspan="6">O&#x2019;Neill et al. (<xref ref-type="bibr" rid="ref5">5</xref>)</td>
</tr>
<tr>
<td align="left" valign="top">Boxer</td>
<td align="left" valign="top">1,386 animals</td>
<td align="left" valign="top">ophthalmic and general disorders</td>
<td align="left" valign="top">Ulcer (4.98%)</td>
</tr>
<tr>
<td align="left" valign="top">Shih Tzu</td>
<td align="left" valign="top">2031 animals</td>
<td align="left" valign="top">ophthalmic and general disorders</td>
<td align="left" valign="top">Ulcer (3.45%)</td>
</tr>
<tr>
<td align="left" valign="top">Cavalier King Charles Spaniel</td>
<td align="left" valign="top">2,332 animals</td>
<td align="left" valign="top">ophthalmic and general disorders</td>
<td align="left" valign="top">Ulcer (2.49%)</td>
</tr>
<tr>
<td align="left" valign="top">Bulldog</td>
<td align="left" valign="top">787 animals</td>
<td align="left" valign="top">ophthalmic and general disorders</td>
<td align="left" valign="top">Ulcer (2.41%)</td>
</tr>
<tr>
<td align="left" valign="top">French Bulldog</td>
<td align="left" valign="top">642 animals</td>
<td align="left" valign="top">ophthalmic and general disorders</td>
<td align="left" valign="top">Ulcer (1.87%)</td>
</tr>
</tbody>
</table>
<table-wrap-foot>
<p>Overview of studies on diseases of ocular surface diseases in brachycephalic dog breeds. For each breed, the number of animals or eyes examined, the general health status (e.g., presence of ocular diseases or healthy) and the most commonly reported ocular surface diseases are summarized. The data show breed-specific patterns in disease prevalence, particularly with regard to keratoconjunctivitis sicca (KCS), corneal ulcers, corneal pigmentation, and stromal fibrosis. The references refer to the original sources for each data set.</p>
</table-wrap-foot>
</table-wrap>
<p>One of the brachycephalic breeds most commonly seen in specialist eye clinics is the French Bulldog. In 2024, 15.2% of all patients in a specialist eye clinic were French Bulldogs (<xref ref-type="bibr" rid="ref14">14</xref>). Not only has their popularity increased in recent years, with registrations at the British Kennel Club rising by about 5.5 times between 2014 and 2021 (<xref ref-type="bibr" rid="ref15">15</xref>), but they were the most popular dog breed in the United States in 2022 (<xref ref-type="bibr" rid="ref16">16</xref>). In a retrospective study of 809 French Bulldogs, 76.6% showed keratitis (spontaneous chronic corneal epithelial defects (SCEED), 31%; melting ulcers, 19%; stromal keratitis, 16.3%; pigmentary keratitis, 5.3%; granulomatous keratitis, 5.2%) and 45% showed tear film disorders (quantitative KCS, 16.2%; qualitative KCS, 28.8%), underscoring the severity and frequency of ocular surface disease in this breed (<xref ref-type="bibr" rid="ref17">17</xref>). Quantitative changes (aqueous deficient dry eye, ADDE) and qualitative changes in the tear film (evaporative dry eye disease, EDED) directly contribute to the development of ocular surface diseases (<xref ref-type="bibr" rid="ref1">1</xref>). We hypothesized that French Bulldogs suffer disproportionately from qualitative tear film disorders.</p>
<p>The main objective of this study was to collect normative data on the characteristics of the ocular surface of French Bulldogs in order to advance research in the field of breed differences. The focus was on the examination of the tear film, which was evaluated using the I.C.P. OSA-Vet&#x00AE; and a series of additional tear film tests. These data were compared with those of non-brachycephalic breeds. Another objective was to investigate the usefulness of measuring osmolarity as an additional parameter for distinguishing between ADDE and EDED in veterinary medicine.</p>
</sec>
<sec sec-type="materials|methods" id="sec2">
<label>2</label>
<title>Materials and methods</title>
<p>The study population of this prospective study consisted of a group of healthy French Bulldogs and a control group of healthy non-brachycephalic breeds. French Bulldogs were included in the study if at least one eye was free of ocular disease and no topical eye medications had been administered within 5&#x202F;days prior to the examination. The dogs were recruited either as inpatients with unilateral eye disease (with examination of the unaffected eye) or as clinically healthy pets from the same household. The control group consisted of mesocephalic and dolichocephalic dogs recruited from routine hereditary eye examinations, patients with unilateral eye disease, or healthy pets of the same household. In all cases, only eyes without clinical abnormalities and without recent topical treatment were included.</p>
<p>The owner&#x2019;s consent for the evaluation of the anonymized data of their dog(s) and the photos taken for research purposes and publication was obtained prior to the examination. All examinations were carried out in accordance with the GERVO Declaration on the Use of Animals in Eye and Vision Research and with the approval of the Berlin State Secretariat for Health and Social Affairs under the number StN 024/22.</p>
<p>All dogs underwent a complete ophthalmic examination, including menace response, dazzle reflex, pupillary light reflex, palpebral reflex, slit lamp biomicroscopy (Kowa SL-17; Kowa, Japan), rebound tonometry (TonoVet, iCare, Finland), indirect ophthalmoscopy (Video Omega 2C; HEINE Optotechnik GmbH &#x0026; Co. KG, Germany), a complete tear film analysis with the I.C.P. OSA-Vet&#x00AE; (SBM Sistemi, Italy), and a series of diagnostic tests for tear film deficiency, including osmolarity measurement, punctate fluorescein staining, lissamine green staining, Schirmer tear test I and Schirmer tear test II (all strips Madhu Instruments Pvt., Ltd., India). In 36 French Bulldogs and all non-brachycephalic dogs, tear film osmolarity was measured using the ScoutPro&#x00AE; osmolarity system (Trukera Medical, Southlake, USA). The tear film analysis with the I.C.P. OSA-Vet&#x00AE; was conducted first, followed by a 10&#x202F;min rest period before the remaining examination.</p>
<sec id="sec3">
<label>2.1</label>
<title>Examination procedure</title>
<p>All examinations were performed by the same examiner (JS) in the same room. At the start of each examination, room temperature and humidity were recorded.</p>
<sec id="sec4">
<label>2.1.1</label>
<title>Lipid layer thickness (LLT)/ interferometry (IF) grading</title>
<p>The interferometry measurement was performed using a short video clip of the eye surface after a blink, which was compared with the supplied video rating scale, taking into account the thickness of the lipid layer and thermodynamics (<xref ref-type="bibr" rid="ref18">18</xref>). The rating scale comprised the following categories, as defined in I.C.P. OSA-Vet&#x00AE;: 0&#x202F;=&#x202F;no lipid layer (lipid layer thickness &#x003C; 15&#x202F;nm), A&#x202F;=&#x202F;open meshwork (lipid layer thickness &#x2248; 15&#x202F;nm), B&#x202F;=&#x202F;closed meshwork (lipid layer thickness &#x2248; 30&#x202F;nm), C&#x202F;=&#x202F;wave pattern (lipid layer thickness &#x2248;30&#x2013;80&#x202F;nm), D&#x202F;=&#x202F;amorphous pattern (lipid layer thickness &#x2248;80&#x202F;nm), E&#x202F;=&#x202F;color fringe (lipid layer thickness &#x2248;80&#x2013;120&#x202F;nm) and F&#x202F;=&#x202F;abnormal color (lipid layer thickness &#x2248;120&#x2013;160&#x202F;nm).</p>
</sec>
<sec id="sec5">
<label>2.1.2</label>
<title>Tear meniscus height (TMH)</title>
<p>The TMH was measured using the I.C.P. OSA-Vet&#x00AE; based on a photo taken after blinking to rule out inaccuracies due to evaporation of the tear film or trichiasis. The upper and lower limits of the tear meniscus were then marked manually on the photo and the TMH was calculated using the software provided. TMH was measured once at the central tear meniscus of the lower eyelid.</p>
</sec>
<sec id="sec6">
<label>2.1.3</label>
<title>Non-invasive breakup time (NIBUT)</title>
<p>The non-invasive tear film break-up time was measured using the I.C.P. OSA-Vet&#x00AE; based on projected Placido rings on the corneal surface. After manually blinking, the eye was kept open to record the video. The software measured the time until the rings deformed (<xref ref-type="bibr" rid="ref18">18</xref>). In cases where automatic measurement was not possible (e.g., due to slight movements of the patient), the time was measured manually in the I.C.P. OSA-Vet&#x00AE; by marking the starting point after blinking and the end point when the first rings were deformed.</p>
</sec>
<sec id="sec7">
<label>2.1.4</label>
<title>Non-contact infrared meibography</title>
<p>The loss of Meibomian glands was measured using the I.C.P. OSA-Vet&#x00AE; by manually everting the tarsal plates of the upper and lower eyelids. Several images were taken and the area of the Meibomian glands was either marked by the software with manual control or marked manually. The software calculated the percentage loss of Meibomian glands (MGL) in relation to the tarsal plate (<xref ref-type="bibr" rid="ref18">18</xref>).</p>
</sec>
<sec id="sec8">
<label>2.1.5</label>
<title>Osmolarity</title>
<p>The measurement was performed using the ScoutPro&#x00AE; osmolarity system on the tear meniscus of the lateral canthus in the lower eyelid area using disposable osmolarity test cards. The device collects 50 nanoliters of tear fluid and measures the osmolarity using a temperature-corrected impedance measurement (<xref ref-type="bibr" rid="ref19">19</xref>). At the beginning of each day of testing, a quality control check was performed using the control card provided by the manufacturer.</p>
</sec>
<sec id="sec9">
<label>2.1.6</label>
<title>Schirmer tear test I (STT-I)</title>
<p>The Schirmer tear test I (Tear Touch Blu&#x00AE;, Madhu Instruments Pvt. Ltd., India) was performed by placing the strip in the lower lateral fornix of the eye. After one minute, the result was measured in mm/min.</p>
</sec>
<sec id="sec10">
<label>2.1.7</label>
<title>Schirmer tear test II (STT-II)</title>
<p>A drop of a topical anesthetic (proxymetacaine hydrochloride; Proparakain-POS 0.5%&#x00AE;, Ursapharm, Germany) was applied to the eye. After one minute, the excess liquid was carefully wiped off with a swab and the Schirmer tear test II was performed as described above for the Schirmer tear test I.</p>
</sec>
<sec id="sec11">
<label>2.1.8</label>
<title>Punctate fluorescein staining (PFS)</title>
<p>Fluorescein staining was performed by instilling a drop of a topical anesthetic (proxymetacaine hydrochloride; Proparakain-POS 0.5%&#x00AE;, Ursapharm, Germany) onto the eye via a fluorescein strip (Fluoro Touch&#x00AE;, Madhu Instruments Pvt., Ltd., India). The fluorescein was distributed by blinking several times manually. Excess fluorescein was washed out with sterile saline solution. The corneal surface was examined with a blue filter and 10x magnification using the Kowa SL 17 slit lamp. Classification was based on the evaluation system for surface area specified by Saito et al. (<xref ref-type="bibr" rid="ref20">20</xref>) whereby grade 0 corresponds to no fluorescein uptake, grade 1 to uptake of less than 50% of the corneal surface, and grade 2 to uptake of more than 50% of the cornea.</p>
</sec>
<sec id="sec12">
<label>2.1.9</label>
<title>Lissamine green staining (LGS)</title>
<p>A lissamine green strip (Green Touch&#x00AE;, Madhu Instruments Pvt., Ltd., India) was moistened with a drop of topical anesthetic and applied once to the dorsal conjunctiva of the eyeball. The dye was distributed by manual blinking. The temporal conjunctival surface was examined using 10x magnification with the Kowa SL-17. Staining was graded as 0&#x202F;=&#x202F;absent and 1&#x202F;=&#x202F;positive. The severity of LGS is associated with reduced STT-I values (<xref ref-type="bibr" rid="ref21">21</xref>) and the diagnostic options for evaporative dry eye are unclear (<xref ref-type="bibr" rid="ref22">22</xref>). Since we did not expect low STT-I values and hypothezised qualitative tear film diosorders in the French Bulldog breed, no further differentiation was made.</p>
</sec>
</sec>
<sec id="sec13">
<label>2.2</label>
<title>Statistical analysis</title>
<p>The statistical analysis of the data was performed by Novustat GmbH (Wollerau, Switzerland). When both eyes of a dog were examined, the results were averaged (<xref ref-type="bibr" rid="ref23">23</xref>). Bilateral differences were negligible and would not have been obscured by averaging. QQ plots indicate that the assumption of normally distributed data was not met. Therefore, the Wilcoxon test was used and <italic>p</italic>&#x202F;&#x003C;&#x202F;0.05 was considered statistically significant. Due to the presence of ties, the implementation of Hothorn et al. was used. In the case of ties, average ranks were calculated (<xref ref-type="bibr" rid="ref24">24</xref>). Besides the uneven sample size, the loss of statistical power due to the unbalanced design is negligible (<xref ref-type="bibr" rid="ref25">25</xref>). Since IF Grade and PFS are measured on an ordinal scale, Spearman&#x2019;s rank correlation coefficient was used. The strength of the correlation was assessed using the following scheme: very weak (0&#x2013;0.19), weak (0.2&#x2013;0.39), moderate (0.40&#x2013;0.59), strong (0.6&#x2013;0.79), very strong (0.8&#x2013;1.0) (<xref ref-type="bibr" rid="ref26">26</xref>).</p>
</sec>
</sec>
<sec sec-type="results" id="sec14">
<label>3</label>
<title>Results</title>
<sec id="sec15">
<label>3.1</label>
<title>Animals</title>
<p>Fifty client-owned French bulldogs (90 eyes) with an average age of 4.96 (0.5&#x2013;12&#x202F;years) and thirty client-owned non-brachycephalic breeds (58 eyes) were included in this study. Among the non-brachycephalic breeds were mixed breeds (n&#x202F;=&#x202F;7), Shetland Sheepdog (n&#x202F;=&#x202F;5), Golden Retriever (n&#x202F;=&#x202F;4), Greyhound (n&#x202F;=&#x202F;3), English Cocker Spaniel (n&#x202F;=&#x202F;2), Polish Lowland Sheepdog (n&#x202F;=&#x202F;2), Siberian Husky (n&#x202F;=&#x202F;1), Saluki (n&#x202F;=&#x202F;1), Greater Swiss Mountain Dog (n&#x202F;=&#x202F;1), Whippet (n&#x202F;=&#x202F;1), Flat-Coated Retriever (n&#x202F;=&#x202F;1), Rough Collie (n&#x202F;=&#x202F;1), Terrier (n&#x202F;=&#x202F;1) and Australian Cattle Dog (n&#x202F;=&#x202F;1). The average age of the non-brachycephalic dogs was 4.88&#x202F;years (0.6&#x2013;10&#x202F;years). The French Bulldogs consisted of 17 females, 10 spayed females, 19 males, and 4 neutered males. The control group consisted of 13 females, 6 spayed females, 6 males, and 5 neutered males.</p>
</sec>
<sec id="sec16">
<label>3.2</label>
<title>Ophthalmic examination</title>
<p>Based on the results of the ophthalmological examination, eight eyes (SCCED n&#x202F;=&#x202F;2; ulceration n&#x202F;=&#x202F;3, eyelid margin mass n&#x202F;=&#x202F;1, dermoid n&#x202F;=&#x202F;1, keratoconjunctivitis sicca n&#x202F;=&#x202F;1) from the French Bulldog group and two eyes (reactive conjunctivitis n&#x202F;=&#x202F;1, eyelid margin mass n&#x202F;=&#x202F;1) from the control group were excluded from the study. Two French Bulldogs were presented after a previous unilateral enucleation.</p>
</sec>
<sec id="sec17">
<label>3.3</label>
<title>Age</title>
<p>Age showed a weak negative correlation with STT-I (<italic>r</italic>&#x202F;=&#x202F;&#x2212;0.32; <italic>p</italic>&#x202F;=&#x202F;0.026) and osmolarity (<italic>r</italic>&#x202F;=&#x202F;&#x2212;0.38; <italic>p</italic>&#x202F;=&#x202F;0.021).</p>
</sec>
<sec id="sec18">
<label>3.4</label>
<title>STT-I and STT-II</title>
<p>The STT-I values were 19.05&#x202F;&#x00B1;&#x202F;4.00&#x202F;mm/min in French Bulldogs and 16.88&#x202F;&#x00B1;&#x202F;3.54&#x202F;mm/min in the control group. French Bulldogs had a significantly higher STT-I value (<italic>p</italic>&#x202F;=&#x202F;0.01) than non-brachycephalic breeds. The STT-II values were 10.16&#x202F;&#x00B1;&#x202F;2.86&#x202F;mm/min in French Bulldogs and 11.28&#x202F;&#x00B1;&#x202F;2.60&#x202F;mm/min in the control group (<xref ref-type="fig" rid="fig1">Figure 1</xref>). The difference was not significant (<italic>p</italic>&#x202F;=&#x202F;0.06). STT-I showed a weak negative correlation (<italic>r</italic>&#x202F;=&#x202F;&#x2212;0.24) with PFS.</p>
<fig position="float" id="fig1">
<label>Figure 1</label>
<caption>
<p>Box plots illustrating the distribution of tear film parameters in French Bulldogs (FB) and non-brachycephalic control dogs. The variables shown include: <bold>(A)</bold> tear osmolarity (mOsm/L); <bold>(B)</bold> tear meniscus height (mm); <bold>(C)</bold> Schirmer tear test I (mm/min); <bold>(D)</bold> non-invasive tear film breakup time (NIBUT; sec.). Each box represents the interquartile range between the 25th and 75th percentiles, with the horizontal line within the box indicating the median. The whiskers extend to the most extreme data points within 1.5 times the IQR, and individual points represent outliers outside this range.</p>
</caption>
<graphic xlink:href="fvets-12-1732237-g001.tif" mimetype="image" mime-subtype="tiff">
<alt-text content-type="machine-generated">Four box plots comparing tear-related measurements in French Bulldogs and a control group. A: Tear Osmolarity shows similar results with p = 0.402. B: Tear Meniscus Height is higher in Bulldogs with p &#x003C; 0.0001. C: Schirmer Tear Test I reveals higher values in Bulldogs with p = 0.0169. D: N.I.B.U.T. is lower in Bulldogs with p &#x003C; 0.0001.</alt-text>
</graphic>
</fig>
</sec>
<sec id="sec19">
<label>3.5</label>
<title>Lipid layer thickness (LLT)/ interferometry (IF) grading</title>
<p>No dog from either group achieved an IF score of F. French Bulldogs showed a significantly lower (<italic>p</italic>&#x202F;=&#x202F;9&#x002A;10<sup>&#x2212;8</sup>) IF score than non-brachycephalic dogs. A comparison between the left and right eyes of both groups is shown in <xref ref-type="fig" rid="fig2">Figure 2</xref>. The IF grade showed a moderate positive correlation (r&#x202F;=&#x202F;0.49) with NIBUT and a moderate negative correlation with the MGL of the upper eyelid (r&#x202F;=&#x202F;&#x2212;0.42) and the lower eyelid (r&#x202F;=&#x202F;&#x2212;0.46) (<xref ref-type="fig" rid="fig3">Figure 3</xref>).</p>
<fig position="float" id="fig2">
<label>Figure 2</label>
<caption>
<p>Distribution of interferometry grades (top row) and punctate fluorescein staining (PFS; bottom row) in the right and left eyes of French Bulldogs and non-brachycephalic control dogs. The interferometry grades represent lipid layer patterns from 0 (absent) to E (color fringe), based on OSA-Vet&#x00AE; classification; grade F (abnormal color pattern) is not displayed as it was not observed in any animal. PFS values are categorized as 0 (no staining), 1 (&#x003C;50% of the corneal surface), and 2 (&#x003E;50% of the corneal surface affected). Each parameter is reported separately for the left and right eye to account for possible unilateral deviations.</p>
</caption>
<graphic xlink:href="fvets-12-1732237-g002.tif" mimetype="image" mime-subtype="tiff">
<alt-text content-type="machine-generated">Bar charts displaying frequency data for interferometry grades and PFS for French Bulldogs and control groups. Top left: left eye interferometry grades, showing higher frequency in grade B for Bulldogs. Top right: right eye interferometry grades, with Bulldogs also peaking at grade B. Bottom left and right: PFS for left and right eyes, both show more Bulldogs in category 1.</alt-text>
</graphic>
</fig>
<fig position="float" id="fig3">
<label>Figure 3</label>
<caption>
<p>Representative I.C.P. OSA-Vet&#x00AE; images. <bold>(A,B)</bold> Interferometry of the precorneal tear film; <bold>(C,D)</bold> tear meniscus height (TMH) at the inferior central meniscus. <bold>A</bold> French Bulldog, 8&#x202F;years old, open meshwork lipid-layer pattern (grade A). <bold>B</bold> Saluki, 7&#x202F;years old, color fringe pattern (grade E). <bold>C</bold> French Bulldog, 9&#x202F;years old, TMH 0.85&#x202F;mm. <bold>D</bold> Greater Swiss Mountain Dog, 8&#x202F;years old, TMH 0.26&#x202F;mm.</p>
</caption>
<graphic xlink:href="fvets-12-1732237-g003.tif" mimetype="image" mime-subtype="tiff">
<alt-text content-type="machine-generated">Four ocular surface images labeled A&#x2013;D. (A) French Bulldog interferometry showing an open-meshwork lipid-layer pattern (grade A). (B) Saluki interferometry showing a color-fringe pattern (grade E). (C) French Bulldog tear meniscus height (TMH) measured at the inferior central meniscus, 0.85 mm (green overlay). (D) Greater Swiss Mountain Dog TMH measurement 0.26 mm. Panels C&#x2013;D illustrate the higher TMH in the French Bulldog compared with the Greater Swiss Mountain Dog.</alt-text>
</graphic>
</fig>
</sec>
<sec id="sec20">
<label>3.6</label>
<title>Tear meniscus height (TMH)</title>
<p>The TMH values were 0.64&#x202F;&#x00B1;&#x202F;0.20&#x202F;mm in French Bulldogs and 0.31&#x202F;&#x00B1;&#x202F;0.22&#x202F;mm in the control group. French Bulldogs had a significantly (<italic>p</italic>&#x202F;=&#x202F;9&#x002A;10<sup>&#x2212;10</sup>) higher TMH value than the control group (<xref ref-type="fig" rid="fig1">Figures 1</xref>, <xref ref-type="fig" rid="fig3">3</xref>).</p>
</sec>
<sec id="sec21">
<label>3.7</label>
<title>Non-invasive breakup time (NIBUT)</title>
<p>The mean NIBUT values were 7.39&#x202F;&#x00B1;&#x202F;3.32&#x202F;s in French Bulldogs and 14.74&#x202F;&#x00B1;&#x202F;5.92&#x202F;s in the non-brachycephalic group. French Bulldogs had a significantly (<italic>p</italic>&#x202F;=&#x202F;3.5&#x202F;&#x00D7;&#x202F;10<sup>&#x2212;9</sup>) lower NIBUT than the non-brachycephalic group. NIBUT showed a strong negative correlation with the MGL (r&#x202F;=&#x202F;&#x2212;0.65) (<xref ref-type="fig" rid="fig1">Figure 1</xref>). In some French Bulldogs, mucous secretions or reduced patient compliance interfered with automatic NIBUT measurement, requiring manual assessment.</p>
</sec>
<sec id="sec22">
<label>3.8</label>
<title>Non-contact infrared meibography</title>
<p>The Meibomian gland loss was measured for the upper eyelids. The mean MGL was 25.32&#x202F;&#x00B1;&#x202F;19.11% in French Bulldogs and 14.57&#x202F;&#x00B1;&#x202F;9.32% in the control group, showing a significant (<italic>p</italic>&#x202F;=&#x202F;0.027) difference between the two groups. Minor difficulties were encountered with meibography. The presence of conjunctival pigmentation in some French Bulldogs (<xref ref-type="fig" rid="fig4">Figure 4</xref>) led to partial coverage of the Meibomian glands, which impaired image sharpness and subsequent analysis.</p>
<fig position="float" id="fig4">
<label>Figure 4</label>
<caption>
<p>Infrared meibography images taken during examination with the I.C.P. OSA-Vet&#x00AE;. Top: Complete view of the Meibomian glands in the upper eyelid. Bottom: Partial obstruction of gland visualization due to pigmentation of the conjunctiva, as is often seen in French Bulldogs.</p>
</caption>
<graphic xlink:href="fvets-12-1732237-g004.tif" mimetype="image" mime-subtype="tiff">
<alt-text content-type="machine-generated">Infrared meibography showing two eyelid images captured with the I.C.P. OSA-Vet. Top: upper eyelid with meibomian glands clearly visible across the tarsal plate. Bottom: meibography with gland visualization partially obscured by conjunctival pigmentation.</alt-text>
</graphic>
</fig>
</sec>
<sec id="sec23">
<label>3.9</label>
<title>Osmolarity</title>
<p>The osmolarity was 296.6&#x202F;&#x00B1;&#x202F;13.6&#x202F;mOsm/L in the French Bulldogs and 299.8&#x202F;&#x00B1;&#x202F;16.1&#x202F;mOsm/L in the control group. The difference was not significant (<italic>p</italic>&#x202F;=&#x202F;0.4) (<xref ref-type="fig" rid="fig1">Figure 1</xref>).</p>
</sec>
<sec id="sec24">
<label>3.10</label>
<title>Punctate fluorescein staining (PFS)</title>
<p>The PFS showed no significant (<italic>p</italic>&#x202F;=&#x202F;0.1) differences between the French Bulldogs and the control group (<xref ref-type="fig" rid="fig2">Figure 2</xref>). A weak negative correlation (<italic>r</italic>&#x202F;=&#x202F;&#x2212;0.38; <italic>p</italic>&#x202F;=&#x202F;0.0075) was found between the PFS and the IF score.</p>
</sec>
<sec id="sec25">
<label>3.11</label>
<title>Lissamine green staining (LGS)</title>
<p>No LGS uptake was detected in the French bulldog group or in the control group.</p>
<p>A tabular overview of the most important parameters is provided in <xref ref-type="table" rid="tab2">Table 2</xref>.</p>
<table-wrap position="float" id="tab2">
<label>Table 2</label>
<caption>
<p>Tear film parameters: French Bulldogs vs Controls.</p>
</caption>
<table frame="hsides" rules="groups">
<thead>
<tr>
<th align="left" valign="top">Variable</th>
<th align="center" valign="top">French Bulldogs (<italic>n</italic>&#x202F;=&#x202F;50)</th>
<th align="center" valign="top">Control group (<italic>n</italic>&#x202F;=&#x202F;30)</th>
<th align="center" valign="top"><italic>p</italic>-Value between the groups</th>
</tr>
</thead>
<tbody>
<tr>
<td align="left" valign="top">STT-I (mm/min)</td>
<td align="char" valign="top" char="&#x00B1;">19.05 &#x00B1; 4.00</td>
<td align="char" valign="top" char="&#x00B1;">16.88 &#x00B1; 3.54</td>
<td align="center" valign="top">0.01</td>
</tr>
<tr>
<td align="left" valign="top">STT-II (mm/min)</td>
<td align="char" valign="top" char="&#x00B1;">10.16 &#x00B1; 2.86</td>
<td align="char" valign="top" char="&#x00B1;">11.28 &#x00B1; 2.60</td>
<td align="center" valign="top">0.06</td>
</tr>
<tr>
<td align="left" valign="top">TMH (mm)</td>
<td align="char" valign="top" char="&#x00B1;">0.64 &#x00B1; 0.20</td>
<td align="char" valign="top" char="&#x00B1;">0.31 &#x00B1; 0.22</td>
<td align="center" valign="top">9&#x00D7;10<sup>&#x2212;10</sup></td>
</tr>
<tr>
<td align="left" valign="top">NIBUT (sec)</td>
<td align="char" valign="top" char="&#x00B1;">7.39 &#x00B1; 3.32</td>
<td align="char" valign="top" char="&#x00B1;">14.74 &#x00B1; 5.92</td>
<td align="center" valign="top">3.5&#x00D7;10<sup>&#x2212;9</sup></td>
</tr>
<tr>
<td align="left" valign="top">Osmolarity (mOsm/L)</td>
<td align="char" valign="top" char="&#x00B1;">296.6 &#x00B1; 13.6&#x002A;</td>
<td align="char" valign="top" char="&#x00B1;">299.8 &#x00B1; 16.1</td>
<td align="center" valign="top">0.4</td>
</tr>
<tr>
<td align="left" valign="top">MGL upper eyelid (%)</td>
<td align="char" valign="top" char="&#x00B1;">25.32 &#x00B1; 19.11</td>
<td align="char" valign="top" char="&#x00B1;">14.56 &#x00B1; 9.32</td>
<td align="center" valign="top">0.027</td>
</tr>
</tbody>
</table>
<table-wrap-foot>
<p>Comparison of tear film parameters between French Bulldogs (<italic>n</italic>&#x202F;=&#x202F;50) and non-brachycephalic control dogs (<italic>n</italic>&#x202F;=&#x202F;30). Values are given as mean &#x00B1; standard deviation. Statistically significant differences between groups were determined using the Wilcoxon test. &#x002A;<italic>n</italic>&#x202F;=&#x202F;36. STT-I, Schirmer tear test I; STT-II, Schirmer tear test II; TMH, tear meniscus height; NIBUT, non-invasive tear film breakup time; MGL, meibomian gland loss.</p>
</table-wrap-foot>
</table-wrap>
</sec>
<sec id="sec26">
<label>3.12</label>
<title>Room temperature and humidity</title>
<p>Room temperature for the French Bulldogs was 22.09&#x202F;&#x00B1;&#x202F;1.93&#x202F;&#x00B0;C and 21.54&#x202F;&#x00B1;&#x202F;1.03 for the control group. There was no statistical difference between the two groups (<italic>p</italic>&#x202F;=&#x202F;0.42). Humidity was 45.44&#x202F;&#x00B1;&#x202F;8.79% for the French Bulldogs and 52.66&#x202F;&#x00B1;&#x202F;5.56 for the control group. The difference was statistically significant with <italic>p</italic>&#x202F;=&#x202F;0.0002. Room temperature and humidity showed a moderate positive correlation with NIBUT (<italic>r</italic>&#x202F;=&#x202F;0.45; <italic>p</italic>&#x202F;=&#x202F;0.001 and <italic>r</italic>&#x202F;=&#x202F;0.54; <italic>p</italic>&#x202F;=&#x202F;0.0001).</p>
</sec>
</sec>
<sec sec-type="discussion" id="sec27">
<label>4</label>
<title>Discussion</title>
<p>To our knowledge, this is the first study to perform a comprehensive tear film analysis exclusively in French Bulldogs. While previous studies have examined certain brachycephalic breeds such as the Shih Tzu (<xref ref-type="bibr" rid="ref8">8</xref>, <xref ref-type="bibr" rid="ref9">9</xref>) in great detail, the French Bulldog, despite being one of the most popular breeds in many countries (<xref ref-type="bibr" rid="ref15">15</xref>, <xref ref-type="bibr" rid="ref16">16</xref>), has not been the subject of systematic tear film investigation to date. A number of studies (<xref ref-type="bibr" rid="ref2">2</xref>, <xref ref-type="bibr" rid="ref4">4</xref>, <xref ref-type="bibr" rid="ref6">6</xref>, <xref ref-type="bibr" rid="ref7">7</xref>, <xref ref-type="bibr" rid="ref27">27</xref>) have included individual French Bulldogs in larger, heterogeneous cohorts. However, these data do not allow conclusions to be drawn about specific breeds. In contrast, the present study provides the first targeted insight into the tear film characteristics of this breed and highlights the clinical relevance of qualitative tear film abnormalities in otherwise ophthalmologically healthy dogs.</p>
<p>In this study, French Bulldogs showed a significantly higher STT-I value (<italic>p</italic>&#x202F;=&#x202F;0.01) compared to non-brachycephalic breeds. The average STT-I values were 19.05&#x202F;&#x00B1;&#x202F;4.00&#x202F;mm/min in French Bulldogs and 16.88&#x202F;&#x00B1;&#x202F;3.54&#x202F;mm/min in the control group. This result contrasts with previous studies, which found STT-I values to be approximately 14% lower in brachycephalic dogs without clinical eye disease compared to non-brachycephalic dogs (<xref ref-type="bibr" rid="ref1">1</xref>, <xref ref-type="bibr" rid="ref28">28</xref>). This discrepancy in these studies was primarily attributed to reduced corneal sensitivity and its negative effect on the afferent part of the tear reflex in brachycephalic breeds (<xref ref-type="bibr" rid="ref1">1</xref>, <xref ref-type="bibr" rid="ref28">28</xref>). However, the results of this study suggest that this trend is not transferable to French Bulldogs. The increased STT-I in this breed is more likely related to increased reflex tear flow than to increased basal tear production. This assumption is supported by simultaneous evidence of a shortened NIBUT, reduced interferometry values, and a significantly increased TMH, all of which indicate EDED (<xref ref-type="bibr" rid="ref1">1</xref>). It has been demonstrated that the process of increased tear evaporation leads to local dehydration of the cornea, activating both nociceptors and cold thermoreceptors in the cornea. This in turn stimulates the tear reflex (<xref ref-type="bibr" rid="ref29">29</xref>, <xref ref-type="bibr" rid="ref30">30</xref>). The STT-I values observed in this study in French Bulldogs were similar to those reported by Li et al. for brachycephalic dogs (19.05&#x202F;&#x00B1;&#x202F;4.00&#x202F;mm/min vs. 18.2&#x202F;&#x00B1;&#x202F;3.9&#x202F;mm/min) (<xref ref-type="bibr" rid="ref27">27</xref>), but lower than previously described for Shih Tzus (22.0&#x202F;&#x00B1;&#x202F;5.5&#x202F;mm/min) (<xref ref-type="bibr" rid="ref9">9</xref>) and other brachycephalic dogs (20.1&#x202F;&#x00B1;&#x202F;3.4&#x202F;mm/min) (<xref ref-type="bibr" rid="ref28">28</xref>). In contrast, STT-I values in the non-brachycephalic group showed a high correlation with data from the dolichocephalic subgroup (16.88&#x202F;&#x00B1;&#x202F;3.54&#x202F;mm/min vs. 16.8&#x202F;&#x00B1;&#x202F;3.4&#x202F;mm/min) from Li et al. (<xref ref-type="bibr" rid="ref27">27</xref>).</p>
<p>A significant increase in TMH was observed in French Bulldogs compared to the control group (0.64&#x202F;&#x00B1;&#x202F;0.20&#x202F;mm vs. 0.31&#x202F;&#x00B1;&#x202F;0.22&#x202F;mm, <italic>p</italic>&#x202F;=&#x202F;9&#x202F;&#x00D7;&#x202F;10<sup>&#x2212;10</sup>), resulting in more than a twofold increase in TMH compared to control values. These results confirm the report by Li et al. on higher TMH in brachycephalic breeds (<xref ref-type="bibr" rid="ref27">27</xref>). An increase in TMH could be a result of an impairment in the tear drainage system (<xref ref-type="bibr" rid="ref31">31</xref>). However, although the tear drainage system of brachycephalic dogs is anatomically different, they consistently have an additional drainage into the oral cavity (<xref ref-type="bibr" rid="ref32">32</xref>), thus supporting a normal tear drainage. The suggested normal range for TMH, measured with the I.C.P. OSA-Vet&#x00AE; in healthy dogs with normal STT-I, normal eyelid shape, and normal tear drainage system, is 0.53&#x202F;&#x00B1;&#x202F;0.11&#x202F;mm (<xref ref-type="bibr" rid="ref18">18</xref>). This value is higher than the data reported by Li et al. of 0.27&#x202F;&#x00B1;&#x202F;0.12&#x202F;mm for mesocephalic breeds and 0.26&#x202F;&#x00B1;&#x202F;0.09&#x202F;mm for dolichocephalic breeds (<xref ref-type="bibr" rid="ref27">27</xref>), 0.41&#x202F;&#x00B1;&#x202F;0.21&#x202F;mm by Kim et al. for Beagles, 0.32&#x202F;&#x00B1;&#x202F;0.14&#x202F;mm (<xref ref-type="bibr" rid="ref33">33</xref>) and 0.36&#x202F;&#x00B1;&#x202F;0.18&#x202F;mm by Sonego et al. in their groups of non-brachycephalic breeds (<xref ref-type="bibr" rid="ref34">34</xref>), the median values measured by Yoon et al. using optical coherence tomography of the anterior segment of the eye (0.32&#x202F;mm and &#x003C;0.32&#x202F;mm) (<xref ref-type="bibr" rid="ref35">35</xref>) and 0.31&#x202F;&#x00B1;&#x202F;0.22&#x202F;mm in this study. A study by Silva et al. found a TMH of 0.59&#x202F;&#x00B1;&#x202F;0.29&#x202F;mm in the right eye and 0.88&#x202F;&#x00B1;&#x202F;0.27&#x202F;mm in the left eye of healthy Shih Tzus (<xref ref-type="bibr" rid="ref36">36</xref>). In a study on qualitative and quantitative tear tests in various brachycephalic breeds, Voitena et al. reported a TMH of 0.51&#x202F;&#x00B1;&#x202F;0.17 (<xref ref-type="bibr" rid="ref37">37</xref>). Zwiauer-Wolfbeisser et al. reported a TMH of 0.5&#x202F;&#x00B1;&#x202F;0.2&#x202F;mm, with the study population consisting of 58.3% brachycephalic breeds treated for distichiasis, making the population incomparable to the other studies (<xref ref-type="bibr" rid="ref38">38</xref>). Peruccio et al. did not disclose the number of dogs and breeds examined (<xref ref-type="bibr" rid="ref18">18</xref>). The available data on TMH shows that TMH is increased in brachycephalic breeds compared to non-brachycephalic breeds (<xref ref-type="bibr" rid="ref27">27</xref>, <xref ref-type="bibr" rid="ref33 ref34 ref35 ref36 ref37 ref38">33&#x2013;38</xref>). Our results suggest that the significant increase in TMH observed in French Bulldogs, combined with the significant decrease in NIBUT and lower interferometry values, provides convincing evidence of underlying EDED in this breed. A substantial variation in reported TMH is observed among studies involving a brachycephalic cohort. This variations includes lower (<xref ref-type="bibr" rid="ref27">27</xref>, <xref ref-type="bibr" rid="ref37">37</xref>) and higher (<xref ref-type="bibr" rid="ref36">36</xref>) values compared to the values reported in our study. This heterogeneity is likely attributable to variations in cohort composition. These findings underscore the importance of establishing breed-specific reference values for tear film diagnostics, particularly in brachycephalic dogs. The NIBUT was significantly lower in French Bulldogs than in the control group (7.39&#x202F;&#x00B1;&#x202F;3.32&#x202F;s. vs. 14.74&#x202F;&#x00B1;&#x202F;5.92&#x202F;s., <italic>p</italic>&#x202F;=&#x202F;3.5&#x002A;10<sup>&#x2212;9</sup>). This result is consistent with other studies comparing brachycephalic breeds with non-brachycephalic breeds (<xref ref-type="bibr" rid="ref27">27</xref>, <xref ref-type="bibr" rid="ref39">39</xref>). The shorter NIBUT could contribute to the higher STT-I, as reported by Faghihi et al. (<xref ref-type="bibr" rid="ref39">39</xref>). The value for the control group was at the lower end of the reported normal range for tear film breakup time (TBUT) in healthy dogs (19.7&#x202F;&#x00B1;&#x202F;5 to 21.53&#x202F;&#x00B1;&#x202F;7.42&#x202F;s.) (<xref ref-type="bibr" rid="ref40">40</xref>). It was lower than the NIBUT in healthy beagles (<xref ref-type="bibr" rid="ref33">33</xref>). Li et al. (<xref ref-type="bibr" rid="ref27">27</xref>) and Voitena et al. (<xref ref-type="bibr" rid="ref37">37</xref>) reported lower than previously described NIBUT values as well. Li et al. discussed that their lower NIBUT values may be due to environmental influences that contribute to a shorter break-up time, especially in the summer months (<xref ref-type="bibr" rid="ref27">27</xref>). As reported in the results section, relative humidity was higher in the control group, whereas room temperature did not differ between groups. Because higher humidity reduces evaporative loss (<xref ref-type="bibr" rid="ref41">41</xref>), this would be expected to prolong NIBUT in the control group and thereby exaggerate the observed between group difference. Although temperature can influence the tear film, temperature-related bias is unlikely in in this study due to comparable temperatures. Since all ambient values were within the typical indoor range (<xref ref-type="bibr" rid="ref42">42</xref>), the influence is likely to be minor and probably not responsible for the general pattern of significantly shorter NIBUT in French Bulldogs. Nevertheless, residual bias cannot be completely ruled out.</p>
<p>The IF grade was significantly lower in French Bulldogs (<italic>p</italic>&#x202F;=&#x202F;9&#x002A;10<sup>&#x2212;8</sup>) than in the control group, with the majority of French Bulldogs achieving an IF grade of B and the control group achieving a grade of E. The IF grade allows a direct estimation of the lipid layer thickness. The lipid layer is the outermost layer of the tear film, which prevents evaporation of the aqueous phase and stabilizes the tear film (<xref ref-type="bibr" rid="ref43">43</xref>). It is produced by the Meibomian glands (<xref ref-type="bibr" rid="ref43">43</xref>). Increased loss of the Meibomian glands or Meibomian gland dysfunction (MGD) leads to a reduction in the lipid layer (<xref ref-type="bibr" rid="ref22">22</xref>) and thus to a low IF grade. This explains the correlation between a higher MGL and a lower NIBUT. A possible change in the composition of meibum due to MGD may also increase evaporative stress and provoke reflex tearing, which contributes to the higher STT-I and TMH, respectively (<xref ref-type="bibr" rid="ref44">44</xref>). Another factor that must be taken into account is allergies, which are common in brachycephalic breeds (<xref ref-type="bibr" rid="ref45">45</xref>). Studies in human medicine have shown that seasonal allergies alter the morphology of the Meibomian glands, promoting a change in lipid secretion and instability of the tear film (<xref ref-type="bibr" rid="ref46">46</xref>, <xref ref-type="bibr" rid="ref47">47</xref>). In accordance with the previously mentioned findings, preliminary data reported by Marques et al. (ESVO 2025) revealed that atopic dogs suffering from allergic conjunctivitis exhibited lower IF grades in comparison with healthy controls. This observation supports to the hypothesis that ocular allergies are related to impairment of the lipid layer.</p>
<p>We have documented a higher MGL value in French Bulldogs, which could partly explain the lower IF grade. However, some patients had both a low IF grade and a low MGL percentage. Consequently, the MGL percentage is not the only factor influencing the IF grade. MGD is a multifactorial disease in which not only the end stage of glandular loss must be considered, but also qualitative and quantitative changes in the secreted lipid content. This may be due to hyposecretory, obstructive, or hypersecretory mechanisms (<xref ref-type="bibr" rid="ref48">48</xref>, <xref ref-type="bibr" rid="ref49">49</xref>). As Ofri et al. have shown, there are differences in the amount of meibum secreted in different dog breeds (<xref ref-type="bibr" rid="ref50">50</xref>). This also explains the prevalence of low IF grades in French Bulldogs. Another possible cause is incomplete closure of the eyelids during blinking, and the lower blink rate often observed in brachycephalic breeds (<xref ref-type="bibr" rid="ref22">22</xref>). The reduced nerve fiber density of the cornea in brachycephalic dogs (<xref ref-type="bibr" rid="ref51">51</xref>) is thought to be the cause of the reduced blink rate in these breeds (<xref ref-type="bibr" rid="ref1">1</xref>). This leads to meibum not being distributed evenly across the cornea, which promotes destabilization of the tear film. Possible MGD in the French Bulldogs requires further investigation.</p>
<p>There are conflicting statements in the human literature as to whether only the upper eyelid (<xref ref-type="bibr" rid="ref52">52</xref>), the lower eyelid (<xref ref-type="bibr" rid="ref53">53</xref>), or both eyelids (<xref ref-type="bibr" rid="ref54">54</xref>, <xref ref-type="bibr" rid="ref55">55</xref>) need to be measured for the correct diagnosis of Meibomian gland disease. Also, in the veterinary literature there is no consensus on which eyelid should be examined using meibography. Kim et al. performed meibography on both eyelids of Beagles and found it difficult to adequately expose the Meibomian gland area of the lower eyelid (<xref ref-type="bibr" rid="ref33">33</xref>). Kitamura et al. did not examine the lower eyelid because this was found to be uncomfortable for their cohort of Shih Tzus (<xref ref-type="bibr" rid="ref56">56</xref>). Li et al. examined the lower eyelids only because the patients showed discomfort during the examination of the upper eyelid (<xref ref-type="bibr" rid="ref27">27</xref>). In our study, we selected the upper eyelid for meibography during the ophthalmological examination, as most dogs were more comfortable with it.</p>
<p>In this study, tear osmolarity was measured using the ScoutPro&#x00AE;. Veterinary clinical validation data for this device are limited (<xref ref-type="bibr" rid="ref57">57</xref>). The osmolarity values of both groups were lower than previously reported normal values of TearLab&#x00AE; and I-PEN&#x00AE; Vet (<xref ref-type="bibr" rid="ref58">58</xref>) and the ScoutPro&#x00AE; (<xref ref-type="bibr" rid="ref57">57</xref>). Although reduced NIBUT and LLT values are typically associated with increased evaporation and increased osmolarity (<xref ref-type="bibr" rid="ref29">29</xref>), the lower osmolarity of the French Bulldogs is presumably a consequence of the significantly increased STT-I and TMH values. This interpretation is supported by Garc&#x00ED;a-Res&#x00FA;a et al., who demonstrated a negative correlation between TMH and osmolarity (<xref ref-type="bibr" rid="ref59">59</xref>). Given the lower osmolarity values in the control group, it can be concluded that a tear reflex triggered by handling can be ruled out, as no significant difference was found between the osmolarity of basal tears and reflex tears (<xref ref-type="bibr" rid="ref60">60</xref>).</p>
<p>The clinical interpretability of osmolarity values remains unclear due to a lack of more published data for the ScoutPro&#x00AE; in dogs. After considering the above factors, it was determined that osmolarity within our specific cohort did not exhibit sensitivity to evaporation-related instability. Our observation shows that there was no difference in osmolarity between French Bulldogs and the control group. Consequently, osmolarity is considered an inadequate single marker for qualitative tear film deficiency in dogs. Further validation of the device and standardization of protocols are deemed necessary.</p>
<p>There were no differences between French Bulldogs and the control group regarding PFS and lissamine green staining. This is to be expected as STT-I was not decreased in animals included in this study. High PFS scores correlate with a lower STT-I (<xref ref-type="bibr" rid="ref20">20</xref>). The weak negative correlation between PFS and IF grade aligns with the general findings, as PFS is regarded as a parameter for ocular health (<xref ref-type="bibr" rid="ref22">22</xref>). Smith et al. reported higher lissamine green staining scores in dogs with lower STT-I values and shorter TBUT (<xref ref-type="bibr" rid="ref21">21</xref>).</p>
<p>When interpreting the results, the limitations of the study must be taken into account. The assessment of the Meibomian glands was limited to non-contact infrared meibography. Image quality was partially limited by anatomical conditions or pigmentation. Although allergies are an important factor in MGD in humans, allergy testing was not performed in French Bulldogs, as this was beyond the scope of this study. All examinations and image analyses were performed by a single examiner without formal assessments of intra- or interobserver variability, which may limit reproducibility and introduce potential observer bias.</p>
</sec>
<sec sec-type="conclusions" id="sec28">
<label>5</label>
<title>Conclusion</title>
<p>This study provides strong evidence that all clinically normal French Bulldogs in this cohort showed consistent indicators of EDED. Despite the absence of obvious clinical symptoms, the dogs showed a combination of shortened NIBUT, reduced LLT, increased TMH, and increased STT-I - characteristic features of a qualitative tear film disease. These findings suggest that EDED is not an isolated occurrence but rather a widespread condition within this breed. It is essential that routine ophthalmic examinations of French Bulldogs consider this underlying pathology.</p>
</sec>
</body>
<back>
<sec sec-type="data-availability" id="sec29">
<title>Data availability statement</title>
<p>The original contributions presented in the study are included in the article/supplementary material, further inquiries can be directed to the corresponding author.</p>
</sec>
<sec sec-type="ethics-statement" id="sec30">
<title>Ethics statement</title>
<p>The animal studies were approved by Berlin State Secretariat for Health and Social Affairs. The studies were conducted in accordance with the local legislation and institutional requirements. Written informed consent was obtained from the owners for the participation of their animals in this study.</p>
</sec>
<sec sec-type="author-contributions" id="sec31">
<title>Author contributions</title>
<p>JS: Conceptualization, Data curation, Formal analysis, Investigation, Methodology, Writing &#x2013; original draft. PS: Formal analysis, Visualization, Writing &#x2013; review &#x0026; editing. AM-L: Methodology, Supervision, Writing &#x2013; review &#x0026; editing. IA: Conceptualization, Formal analysis, Funding acquisition, Resources, Supervision, Writing &#x2013; review &#x0026; editing, Methodology.</p>
</sec>
<ack>
<title>Acknowledgments</title>
<p>We would like to express our gratitude to the DOK for their support in conducting this study.</p>
</ack>
<sec sec-type="COI-statement" id="sec32">
<title>Conflict of interest</title>
<p>The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.</p>
</sec>
<sec sec-type="ai-statement" id="sec33">
<title>Generative AI statement</title>
<p>The authors declare that no Gen AI was used in the creation of this manuscript.</p>
<p>Any alternative text (alt text) provided alongside figures in this article has been generated by Frontiers with the support of artificial intelligence and reasonable efforts have been made to ensure accuracy, including review by the authors wherever possible. If you identify any issues, please contact us.</p>
</sec>
<sec sec-type="disclaimer" id="sec34">
<title>Publisher&#x2019;s note</title>
<p>All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.</p>
</sec>
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<fn-group>
<fn fn-type="custom" custom-type="edited-by" id="fn0001">
<p>Edited by: <ext-link ext-link-type="uri" xlink:href="https://loop.frontiersin.org/people/581737/overview">Alberto Mu&#x00F1;oz</ext-link>, University of Murcia, Spain</p>
</fn>
<fn fn-type="custom" custom-type="reviewed-by" id="fn0002">
<p>Reviewed by: <ext-link ext-link-type="uri" xlink:href="https://loop.frontiersin.org/people/1743451/overview">Nirajkumar Makadiya</ext-link>, Santa Rita Animal Care Clinic, United States</p>
<p><ext-link ext-link-type="uri" xlink:href="https://loop.frontiersin.org/people/3262089/overview">J&#x00E9;ssica Voitena</ext-link>, Universidade Federal do Paran&#x00E1;, Brazil</p>
</fn>
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