Utilizing C-Reactive Protein as a Predictor for Subclinical Mastitis

Utilizing C-Reactive Protein as a Predictor for Subclinical Mastitis | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Short Report Utilizing C-Reactive Protein as a Predictor for Subclinical Mastitis Dr. Thiyagarajan Sanjeevi, Valarmathi Sivaraman, Yogasri Karunanithi This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-5780805/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Mastitis stands out as the most economically significant ailment in dairy cows due to its adverse effects on milk quality and reproductive performance, often leading to the involuntary culling of affected cows. Subclinical mastitis (SCM) can be considered a precursor to severe mastitis. This current study sheds light on the role of C-reactive protein (CRP) as a potential indicator for the presence of bovine SCM. The research involved the analysis of milk samples from a total of 49 cows, encompassing crossbred and native breeds. These samples were evaluated for their CRP levels and then compared with the absolute somatic cell count. To identify SCM, a somatic cell count within the range of 5.00-7.5 lakhs/ml was used as a threshold. The results revealed elevated CRP concentrations in the milk of cows afflicted with clinical mastitis (11.72 ± 0.75 µg/mL) as well as those with subclinical mastitis (10.9 ± 1.2 µg/mL), in stark contrast to the levels observed in healthy cows (0.33 ± 0.02 µg/mL). This research suggests that CRP may serve as a valuable tool for the ongoing surveillance of bovine mastitis, ultimately contributing to improved milk production performance. Biochemical Research Methods C-Reactive protein Milk bovine subclinical mastitis Figures Figure 1 INTRODUCTION Bovine mastitis is an inflammatory condition affecting the mammary glands, often characterized by milk stagnation in the glandular tissue. This condition is primarily observed during the lactation period, especially in high-yielding breeds. It typically leads to an ascending infection within the lactating gland, spreading through the bloodstream. The onset of mastitis can be categorized as clinical or subclinical, depending on the presence of clinical symptoms. It is a matter of concern due to its potential to progress to sepsis, septic shock, and fibrosis in later stages [ 1 ]. Therefore, there is a pressing need for early and accurate diagnosis of mastitis in bovines. Presently, clinical bovine mastitis is primarily diagnosed through physical examination and additional tests, including pH measurement, conductivity assessment, milk microbial analysis, and somatic cell counts [ 2 ]. However, more sensitive and non-invasive biomarkers are required to enhance early diagnosis and facilitate effective therapy monitoring. The assessment of acute-phase proteins (APPs) in milk has been proposed as the most sensitive and non-invasive method for detecting bovine mastitis [ 3 ]. Despite this, the use of C-reactive protein (CRP) as a predictive marker for mastitis has not been extensively explored. CRP is a valuable indicator of inflammation and treatment progress, as its serum levels rapidly rise following tissue damage and decrease with successful therapy [ 4 ]. Therefore, we hypothesized that CRP could be measured in bovine milk samples and might exhibit elevated levels in cases of mastitis. To test this hypothesis, we estimated the CRP concentrations in random milk samples and categorized the milk as subclinical or mastitis based on the somatic cell counts within. Additionally, we explored the correlation between milk CRP concentrations and somatic cell counts in both healthy and diseased lactating animals. MATERIALS AND METHODS Random milk samples (n = 49) were collected directly from the udder of milking cows from Goh sala and various private farms in and around Puducherry. To count the somatic cells in the milk, the glass slides with milk smears were placed on the slide rack and were flooded with modified Newman-Lampert stain (Himedia) for 2 min. The excess stain was drained off by standing the slices on absorbent paper; slides were rinsed thrice in tap water and air-dried. The slides were examined under oil immersion objective and the number of cells in 20 fields was counted and the total number of cells per ml of milk was calculated [ 5 ]. Milk CRP was determined using a commercial assay kit (Regenix lab, Chennai). Before analysis, the milk samples were centrifuged (10000 × g, 30 min, 4°C) to remove fat and the milk samples were diluted 1:200 in assay buffer (Tris–HCl buffered NaCl solution (pH 7.8), containing < 0.1% NaN3, Tween 40, and diethylene-triaminepentaaceticacid. The assay was performed as per the manufacturer’s instructions. RESULTS AND DISCUSSION The findings indicated that within the bovine milk samples, C-reactive protein (CRP) concentrations ranged from 0.2 to 0.42 µg/mL in healthy animals, and from 0.55 to 15.0 µg/mL in diseased ones (P < 0.01). Please refer to Fig. 1 for a graphical representation of the milk CRP data in healthy cows and those with clinical and subclinical mastitis. Notably, cows affected by clinical mastitis exhibited significantly higher milk CRP concentrations at 11.72 ± 0.75 µg/mL, while those with subclinical mastitis had concentrations of 10.9 ± 1.2 µg/mL when compared to the 0.33 ± 0.02 µg/mL in healthy cows. Importantly, there was no statistically significant difference in milk CRP concentrations between cows with clinical and subclinical mastitis. The early detection of bovine mastitis holds paramount clinical significance as it helps mitigate poor reproductive performance and economic losses. CRP serves as a non-specific acute-phase protein, with elevated levels indicating the presence of inflammation of any origin. In the context of bovine mastitis, increased milk CRP levels signify ongoing inflammation in the mammary gland [ 6 ]. However, it's worth noting that the diversity of pathologies involved in clinical mastitis could explain the observed high data variability. Conversely, reports have suggested that early diagnosis in clinically active cases might not detect increased CRP concentrations, as CRP levels are known to reach their peak concentrations 24 hours after tissue damage [ 7 ]. Consequently, based on findings from experimental subclinical mastitis in cows [ 3 ], it can be anticipated that increased CRP levels would be evident in severe subclinical mastitis cases. Nevertheless, further studies involving a larger number of animals are warranted to validate these observations. CONCLUSIONS The data from the current study reveals that milk C-reactive protein (CRP) levels were elevated in affected bovines, signifying the presence of both local and systemic inflammation in mastitis cases. Furthermore, a positive correlation was observed between CRP concentrations and the somatic cell count. However, it's important to note that there were no statistically significant differences in milk CRP concentrations between clinical and subclinical mastitis. One of the limitations of this research pertains to the relatively small sample size, which precluded the derivation of comprehensive conclusions. Additionally, due to the random nature of many of the sampled milking sessions, the exact timing of illness onset could not be recorded and analyzed in this study. Nevertheless, the results provide a genuine clinical representation, offering promising data that warrants further investigation into this subject matter. Declarations CONFLICTS OF INTEREST There are no conflicts of interest. ACKNOWLEDGEMENT The authors are highly grateful to the management of Aarupadai Veedu Medical College and Hospital, Vinayaka Missions Puducherry Campus and Translational Research Platform for Veterinary Biological, Tamilnadu Veterinary and Animal Science University, Chennai for technical supports and owners of dairy farms for providing milk samples. References Adkin PRF, Middleton JR,2018. Methods for diagnosingmastitis. VeterinaryClinics of North Food AnimalPractice , 34(3), 479–491. doi:10.1016/j.cvfa.2018.07.003 Ashraf A, Imran M, 2018. Diagnosis of bovine mastitis: From laboratory to farm. TropicalAnimal Health and Production,50(6), 1193–1202. doi:1007/s11250-018-1629-0 Eckersall PD, Young FJ, McComb C, 2001. Acute phase proteins in serum and milk from dairy cows with clinical mastitis. VeterinaryRecord,148(2), 35–41. doi:1136/vr.148.2.35 Jain S, Gautam V, Naseem S, 2011. Acute-phase proteins: As diagnostic tool. Journal of Pharmacy and Bioallied Science s , 3(1), 118–127. doi:4103/0975-7406.76489 Petersson KH, Connor LA, Petersson-Wolfe CS, et al, 2011. Evaluation of confirmatory stains used for direct microscopic somatic cell counting of sheep milk. Journal of Dairy Science,94(4), 1908–1912. doi:3168/jds.2010-3574 Thomas FC, Geraghty T, Simões PBA, 2018. A pilot study of acute phase proteins as indicators of bovine mastitis caused by different pathogens. Research in VeterinaryScience,119, 176–181. doi:1016/j.rvsc.2018.06.015 Dalanezi FM, Schmidt EMS, Joaquim SF, et al, 2020. Concentrations of acute-phaseproteins in milk from cows with clinicalmastitiscaused by differentpathogens. Pathogens,9(9), 706. doi:3390/pathogens9090706 Additional Declarations The authors declare no competing interests. Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-5780805","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Short Report","associatedPublications":[],"authors":[{"id":398839851,"identity":"e7dce7fe-98ec-431d-9c3f-d833674ca3f2","order_by":0,"name":"Dr. Thiyagarajan Sanjeevi","email":"data:image/png;base64,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","orcid":"https://orcid.org/0000-0002-4006-7184","institution":"Department of Medical Biotechnology, Multi-Disciplinary Center for Biomedical Research, Aarupadai Veedu Medical College and Hospital, Vinayaka Missions Research Foundation -DU, Puducherry. 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This condition is primarily observed during the lactation period, especially in high-yielding breeds. It typically leads to an ascending infection within the lactating gland, spreading through the bloodstream. The onset of mastitis can be categorized as clinical or subclinical, depending on the presence of clinical symptoms. It is a matter of concern due to its potential to progress to sepsis, septic shock, and fibrosis in later stages [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e]. Therefore, there is a pressing need for early and accurate diagnosis of mastitis in bovines.\u003c/p\u003e \u003cp\u003ePresently, clinical bovine mastitis is primarily diagnosed through physical examination and additional tests, including pH measurement, conductivity assessment, milk microbial analysis, and somatic cell counts [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e]. However, more sensitive and non-invasive biomarkers are required to enhance early diagnosis and facilitate effective therapy monitoring. The assessment of acute-phase proteins (APPs) in milk has been proposed as the most sensitive and non-invasive method for detecting bovine mastitis [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eDespite this, the use of C-reactive protein (CRP) as a predictive marker for mastitis has not been extensively explored. CRP is a valuable indicator of inflammation and treatment progress, as its serum levels rapidly rise following tissue damage and decrease with successful therapy [\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e]. Therefore, we hypothesized that CRP could be measured in bovine milk samples and might exhibit elevated levels in cases of mastitis. To test this hypothesis, we estimated the CRP concentrations in random milk samples and categorized the milk as subclinical or mastitis based on the somatic cell counts within. Additionally, we explored the correlation between milk CRP concentrations and somatic cell counts in both healthy and diseased lactating animals.\u003c/p\u003e"},{"header":"MATERIALS AND METHODS","content":"\u003cp\u003eRandom milk samples (n\u0026thinsp;=\u0026thinsp;49) were collected directly from the udder of milking cows from Goh sala and various private farms in and around Puducherry. To count the somatic cells in the milk, the glass slides with milk smears were placed on the slide rack and were flooded with modified Newman-Lampert stain (Himedia) for 2 min. The excess stain was drained off by standing the slices on absorbent paper; slides were rinsed thrice in tap water and air-dried. The slides were examined under oil immersion objective and the number of cells in 20 fields was counted and the total number of cells per ml of milk was calculated [\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e]. Milk CRP was determined using a commercial assay kit (Regenix lab, Chennai). Before analysis, the milk samples were centrifuged (10000 \u0026times; g, 30 min, 4\u0026deg;C) to remove fat and the milk samples were diluted 1:200 in assay buffer (Tris\u0026ndash;HCl buffered NaCl solution (pH 7.8), containing\u0026thinsp;\u0026lt;\u0026thinsp;0.1% NaN3, Tween 40, and diethylene-triaminepentaaceticacid. The assay was performed as per the manufacturer\u0026rsquo;s instructions.\u003c/p\u003e"},{"header":"RESULTS AND DISCUSSION","content":"\u003cp\u003eThe findings indicated that within the bovine milk samples, C-reactive protein (CRP) concentrations ranged from 0.2 to 0.42 \u0026micro;g/mL in healthy animals, and from 0.55 to 15.0 \u0026micro;g/mL in diseased ones (P\u0026thinsp;\u0026lt;\u0026thinsp;0.01). Please refer to Fig.\u0026nbsp;1 for a graphical representation of the milk CRP data in healthy cows and those with clinical and subclinical mastitis.\u003c/p\u003e \u003cp\u003e Notably, cows affected by clinical mastitis exhibited significantly higher milk CRP concentrations at 11.72\u0026thinsp;\u0026plusmn;\u0026thinsp;0.75 \u0026micro;g/mL, while those with subclinical mastitis had concentrations of 10.9\u0026thinsp;\u0026plusmn;\u0026thinsp;1.2 \u0026micro;g/mL when compared to the 0.33\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02 \u0026micro;g/mL in healthy cows. Importantly, there was no statistically significant difference in milk CRP concentrations between cows with clinical and subclinical mastitis.\u003c/p\u003e \u003cp\u003eThe early detection of bovine mastitis holds paramount clinical significance as it helps mitigate poor reproductive performance and economic losses. CRP serves as a non-specific acute-phase protein, with elevated levels indicating the presence of inflammation of any origin. In the context of bovine mastitis, increased milk CRP levels signify ongoing inflammation in the mammary gland [\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]. However, it's worth noting that the diversity of pathologies involved in clinical mastitis could explain the observed high data variability. Conversely, reports have suggested that early diagnosis in clinically active cases might not detect increased CRP concentrations, as CRP levels are known to reach their peak concentrations 24 hours after tissue damage [\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e]. Consequently, based on findings from experimental subclinical mastitis in cows [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e], it can be anticipated that increased CRP levels would be evident in severe subclinical mastitis cases. Nevertheless, further studies involving a larger number of animals are warranted to validate these observations.\u003c/p\u003e"},{"header":"CONCLUSIONS","content":"\u003cp\u003eThe data from the current study reveals that milk C-reactive protein (CRP) levels were elevated in affected bovines, signifying the presence of both local and systemic inflammation in mastitis cases. Furthermore, a positive correlation was observed between CRP concentrations and the somatic cell count. However, it's important to note that there were no statistically significant differences in milk CRP concentrations between clinical and subclinical mastitis.\u003c/p\u003e \u003cp\u003eOne of the limitations of this research pertains to the relatively small sample size, which precluded the derivation of comprehensive conclusions. Additionally, due to the random nature of many of the sampled milking sessions, the exact timing of illness onset could not be recorded and analyzed in this study. Nevertheless, the results provide a genuine clinical representation, offering promising data that warrants further investigation into this subject matter.\u003c/p\u003e"},{"header":"Declarations","content":" \u003cp\u003e \u003cstrong\u003eCONFLICTS OF INTEREST\u003c/strong\u003e \u003cp\u003eThere are no conflicts of interest.\u003c/p\u003e \u003c/p\u003e\u003ch2\u003eACKNOWLEDGEMENT\u003c/h2\u003e \u003cp\u003eThe authors are highly grateful to the management of Aarupadai Veedu Medical College and Hospital, Vinayaka Missions Puducherry Campus and Translational Research Platform for Veterinary Biological, Tamilnadu Veterinary and Animal Science University, Chennai for technical supports and owners of dairy farms for providing milk samples.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eAdkin PRF, Middleton JR,2018. Methods for diagnosingmastitis. VeterinaryClinics of North Food AnimalPractice\u003cem\u003e,\u003c/em\u003e 34(3), 479\u0026ndash;491. doi:10.1016/j.cvfa.2018.07.003\u003c/li\u003e\n\u003cli\u003eAshraf A, Imran M, 2018. Diagnosis of bovine mastitis: From laboratory to farm. TropicalAnimal Health and Production,50(6), 1193\u0026ndash;1202. doi:1007/s11250-018-1629-0\u003c/li\u003e\n\u003cli\u003eEckersall PD, Young FJ, McComb C, 2001. Acute phase proteins in serum and milk from dairy cows with clinical mastitis. VeterinaryRecord,148(2), 35\u0026ndash;41. doi:1136/vr.148.2.35\u003c/li\u003e\n\u003cli\u003eJain S, Gautam V, Naseem S, 2011. Acute-phase proteins: As diagnostic tool. Journal of Pharmacy and Bioallied Science\u003cem\u003es\u003c/em\u003e, 3(1), 118\u0026ndash;127. doi:4103/0975-7406.76489\u003c/li\u003e\n\u003cli\u003ePetersson KH, Connor LA, Petersson-Wolfe CS, et al, 2011. Evaluation of confirmatory stains used for direct microscopic somatic cell counting of sheep milk. Journal of Dairy Science,94(4), 1908\u0026ndash;1912. doi:3168/jds.2010-3574\u003c/li\u003e\n\u003cli\u003eThomas FC, Geraghty T, Sim\u0026otilde;es PBA, 2018. A pilot study of acute phase proteins as indicators of bovine mastitis caused by different pathogens. Research in VeterinaryScience,119, 176\u0026ndash;181. doi:1016/j.rvsc.2018.06.015\u003c/li\u003e\n\u003cli\u003eDalanezi FM, Schmidt EMS, Joaquim SF, et al, 2020. Concentrations of acute-phaseproteins in milk from cows with clinicalmastitiscaused by differentpathogens. Pathogens,9(9), 706. doi:3390/pathogens9090706\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"hideJournal":true,"highlight":"","institution":"Aarupadai Veedu Medical College \u0026 Hospital","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"C-Reactive protein, Milk, bovine, subclinical, mastitis","lastPublishedDoi":"10.21203/rs.3.rs-5780805/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-5780805/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eMastitis stands out as the most economically significant ailment in dairy cows due to its adverse effects on milk quality and reproductive performance, often leading to the involuntary culling of affected cows. Subclinical mastitis (SCM) can be considered a precursor to severe mastitis. This current study sheds light on the role of C-reactive protein (CRP) as a potential indicator for the presence of bovine SCM. The research involved the analysis of milk samples from a total of 49 cows, encompassing crossbred and native breeds. These samples were evaluated for their CRP levels and then compared with the absolute somatic cell count. To identify SCM, a somatic cell count within the range of 5.00-7.5 lakhs/ml was used as a threshold. The results revealed elevated CRP concentrations in the milk of cows afflicted with clinical mastitis (11.72\u0026thinsp;\u0026plusmn;\u0026thinsp;0.75 \u0026micro;g/mL) as well as those with subclinical mastitis (10.9\u0026thinsp;\u0026plusmn;\u0026thinsp;1.2 \u0026micro;g/mL), in stark contrast to the levels observed in healthy cows (0.33\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02 \u0026micro;g/mL). This research suggests that CRP may serve as a valuable tool for the ongoing surveillance of bovine mastitis, ultimately contributing to improved milk production performance.\u003c/p\u003e","manuscriptTitle":"Utilizing C-Reactive Protein as a Predictor for Subclinical Mastitis","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-01-08 08:27:52","doi":"10.21203/rs.3.rs-5780805/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"7d99a373-ebcd-4c42-afeb-02eb02bd191b","owner":[],"postedDate":"January 8th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":42491534,"name":"Biochemical Research Methods"}],"tags":[],"updatedAt":"2025-01-08T08:27:52+00:00","versionOfRecord":[],"versionCreatedAt":"2025-01-08 08:27:52","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-5780805","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-5780805","identity":"rs-5780805","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}