{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,6]],"date-time":"2026-04-06T11:11:31Z","timestamp":1775473891809,"version":"3.50.1"},"reference-count":37,"publisher":"MDPI AG","issue":"10","license":[{"start":{"date-parts":[[2022,9,23]],"date-time":"2022-09-23T00:00:00Z","timestamp":1663891200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Massey University School of Veterinary Science Postgraduate Research Fund"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biology"],"abstract":"<jats:p>Numerous cases of spontaneous humeral fracture in primiparous dairy cows from New Zealand have prompted the study of the condition to establish probable causes or risk factors associated with the condition. Previous studies identified inadequate protein-calorie malnutrition as an important contributory factor. Earlier case studies also reported that ~50% of cows have low liver and\/or serum copper concentration at the time of humeral fracture. Because copper is so closely associated with the formation of collagen cross-links, the aim of this study was to compare collagen and collagen crosslink content in the humerus from primiparous cows with and without humeral fractures and to determine the role of copper in the occurrence of these fractures. Humeri were collected from cows with and without humeral fractures, ground, and the collagen and collagen cross-link content measured using high-performance liquid chromatography. Collagen content was significantly higher in the humeri of cows without humeral fractures, while total collagen crosslink content was significantly higher in the humerus of cows with humeral fractures. These results indicate other factor\/s (e.g., protein-calorie undernutrition) might be more important than the copper status in the occurrence of humeral fractures in dairy cows in New Zealand.<\/jats:p>","DOI":"10.3390\/biology11101387","type":"journal-article","created":{"date-parts":[[2022,9,25]],"date-time":"2022-09-25T21:14:28Z","timestamp":1664140468000},"page":"1387","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":8,"title":["Novel Assessment of Collagen and Its Crosslink Content in the Humerus from Primiparous Dairy Cows with Spontaneous Humeral Fractures Due to Osteoporosis from New Zealand"],"prefix":"10.3390","volume":"11","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-3515-9164","authenticated-orcid":false,"given":"Alvaro","family":"Wehrle-Martinez","sequence":"first","affiliation":[{"name":"School of Veterinary Sciences, Massey University, Private Bag 11-222, Palmerston North 4442, New Zealand"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2763-5922","authenticated-orcid":false,"given":"Rafea","family":"Naffa","sequence":"additional","affiliation":[{"name":"Fonterra Research and Development Centre, Palmerston North 4472, New Zealand"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8939-686X","authenticated-orcid":false,"given":"Penny","family":"Back","sequence":"additional","affiliation":[{"name":"School of Veterinary Sciences, Massey University, Private Bag 11-222, Palmerston North 4442, New Zealand"},{"name":"School of Agriculture and Environment, Massey University, Private Bag 11-222, Palmerston North 4442, New Zealand"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4253-1825","authenticated-orcid":false,"given":"Chris W.","family":"Rogers","sequence":"additional","affiliation":[{"name":"School of Veterinary Sciences, Massey University, Private Bag 11-222, Palmerston North 4442, New Zealand"},{"name":"School of Agriculture and Environment, Massey University, Private Bag 11-222, Palmerston North 4442, New Zealand"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2453-1485","authenticated-orcid":false,"given":"Kevin","family":"Lawrence","sequence":"additional","affiliation":[{"name":"School of Veterinary Sciences, Massey University, Private Bag 11-222, Palmerston North 4442, New Zealand"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0501-5522","authenticated-orcid":false,"given":"Trevor","family":"Loo","sequence":"additional","affiliation":[{"name":"School of Natural Sciences, Massey University, Private Bag 11-222, Palmerston North 4442, New Zealand"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9485-1428","authenticated-orcid":false,"given":"Andrew","family":"Sutherland-Smith","sequence":"additional","affiliation":[{"name":"School of Natural Sciences, Massey University, Private Bag 11-222, Palmerston North 4442, New Zealand"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1813-2197","authenticated-orcid":false,"given":"Keren","family":"Dittmer","sequence":"additional","affiliation":[{"name":"School of Veterinary Sciences, Massey University, Private Bag 11-222, Palmerston North 4442, New Zealand"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2022,9,23]]},"reference":[{"key":"ref_1","unstructured":"Weston, J., Thompson, K., Dittmer, K., and Rashid, Z. (2012, January 19\u201324). Humeral fractures in dairy heifers. Proceedings of the Dairy Cattle Veterinarians of the New Zealand Veterinary Association, Hamilton, New Zealand."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"230","DOI":"10.1080\/00480169.2016.1171173","article-title":"Pathophysiology of humeral fractures in a sample of dairy heifers","volume":"64","author":"Dittmer","year":"2016","journal-title":"N. Z. Vet. J."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"23","DOI":"10.3390\/ruminants1010002","article-title":"The Mid-Diaphysis Is a Poor Predictor of Humeral Fracture Risk Indicating That Predisposing Factors Are Recent","volume":"1","author":"Gibson","year":"2021","journal-title":"Ruminants"},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Wehrle-Martinez, A., Lawrence, K., Back, P., Rogers, C., Gibson, M., and Dittmer, K. (2022). Osteoporosis is the cause of spontaneous humeral fracture in dairy cows from New Zealand. Vet. Pathol.","DOI":"10.1177\/03009858221122500"},{"key":"ref_5","unstructured":"Weston, J. (2008). An Outbreak of Humeral Fractures among Dairy Heifers, Society of Dairy Cattle Veterinarians of the New Zealand Veterinary Association."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"148","DOI":"10.1007\/s12011-018-1578-5","article-title":"Copper and zinc nutritional issues for agricultural animal production","volume":"188","author":"Hill","year":"2019","journal-title":"Biol. Trace Elem. Res."},{"key":"ref_7","unstructured":"Constable, P., Hinchcliff, K., Done, S., and Gr\u00fcnberg, W. (2016). Diseases of the Musculoskeletal System. Veterinary Medicine: A Textbook of the Diseases of Cattle, Horses, Sheep, Pigs and Goats, Elsevier Health Sciences. [11th ed.]."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"525","DOI":"10.1016\/j.cvfa.2020.08.009","article-title":"Evaluating Mineral Status in Ruminant Livestock","volume":"36","author":"Ensley","year":"2020","journal-title":"Vet. Clin. N. Am. Food Anim. Pract."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"756","DOI":"10.2460\/javma.2001.218.756","article-title":"Sensitivity and specificity of serum copper determination for detection of copper deficiency in feeder calves","volume":"218","author":"Tessman","year":"2001","journal-title":"J. Am. Vet. Med. Assoc."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"182","DOI":"10.1038\/bonekey.2012.182","article-title":"The contribution of collagen crosslinks to bone strength","volume":"1","author":"Garnero","year":"2012","journal-title":"BoneKey Rep."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"195","DOI":"10.1007\/s00198-009-1066-z","article-title":"Collagen cross-links as a determinant of bone quality: A possible explanation for bone fragility in aging, osteoporosis, and diabetes mellitus","volume":"21","author":"Saito","year":"2010","journal-title":"Osteoporos. Int."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"319","DOI":"10.1007\/s00198-005-2035-9","article-title":"The role of collagen in bone strength","volume":"17","author":"Garnero","year":"2006","journal-title":"Osteoporos. Int."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"242","DOI":"10.1007\/s00223-015-9985-5","article-title":"Effects of collagen crosslinking on bone material properties in health and disease","volume":"97","author":"Saito","year":"2015","journal-title":"Calcif. Tissue Int."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"708","DOI":"10.1093\/jn\/112.4.708","article-title":"Role of copper in collagen cross-linking and its influence on selected mechanical properties of chick bone and tendon","volume":"112","author":"Opsahl","year":"1982","journal-title":"J. Nutr."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"57","DOI":"10.1093\/jn\/98.1.57","article-title":"Effect of copper deficiency on chick bone collagen and selected bone enzymes","volume":"98","author":"Rucker","year":"1969","journal-title":"J. Nutr."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"509","DOI":"10.1016\/j.ijbiomac.2019.01.151","article-title":"Molecular and structural insights into skin collagen reveals several factors that influence its architecture","volume":"128","author":"Naffa","year":"2019","journal-title":"Int. J. Biol. Macromol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"60","DOI":"10.1016\/j.chroma.2016.11.060","article-title":"Liquid chromatography-electrospray ionization mass spectrometry for the simultaneous quantitation of collagen and elastin crosslinks","volume":"1478","author":"Naffa","year":"2016","journal-title":"J. Chromatogr. A"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"225","DOI":"10.1016\/0009-9120(96)00003-6","article-title":"A simplified method for the analysis of hydroxyproline in biological tissues","volume":"29","author":"Reddy","year":"1996","journal-title":"Clin. Biochem."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"299","DOI":"10.1016\/S0021-9258(19)51149-8","article-title":"The determination of hydroxyproline","volume":"184","author":"Neuman","year":"1950","journal-title":"J. Biol. Chem."},{"key":"ref_20","unstructured":"Grace, N., Knowles, S., and Sykes, A. (2010). Copper. Managing Mineral Deficiencies in Grazing Livestock, New Zealand Society of Animal Production."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1186\/1751-0473-3-17","article-title":"Purposeful selection of variables in logistic regression","volume":"3","author":"Bursac","year":"2008","journal-title":"Source Code Biol. Med."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1316","DOI":"10.1177\/1535370216648805","article-title":"Copper, iron, and selenium dietary deficiencies negatively impact skeletal integrity: A review","volume":"241","author":"Medeiros","year":"2016","journal-title":"Exp. Biol. Med."},{"key":"ref_23","first-page":"682","article-title":"Abnormal bone development and lameness associated with secondary copper deficiency in young cattle","volume":"166","author":"Smith","year":"1975","journal-title":"J. Am. Vet. Med. Assoc."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"309","DOI":"10.1079\/BJN19760039","article-title":"Biochemical and pathological changes in tissues of Friesian cattle during the experimental induction of copper deficiency","volume":"35","author":"Mills","year":"1976","journal-title":"Br. J. Nutr."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"137","DOI":"10.1016\/0021-9975(78)90070-1","article-title":"Effects of experimental copper deficiency on the skeleton of the calf","volume":"88","author":"Suttle","year":"1978","journal-title":"J. Comp. Pathol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"2789","DOI":"10.1039\/D0QO00624F","article-title":"Enzymatic and non-enzymatic crosslinks found in collagen and elastin and their chemical synthesis","volume":"7","author":"Gaar","year":"2020","journal-title":"Org. Chem. Front."},{"key":"ref_27","unstructured":"(2012). Quarterly Review of Diagnostic Cases: October to December 2011."},{"key":"ref_28","unstructured":"(2014). Quarterly Report of Diagnostic Cases: October to December 2013."},{"key":"ref_29","unstructured":"(2018). Quarterly Review of Diagnostic Cases: October to December 2017."},{"key":"ref_30","unstructured":"(2019). Quarterly Report of Diagnostic Cases: July to September 2019."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"1045","DOI":"10.1042\/bj3101045","article-title":"Biochemical changes in the collagenous matrix of osteoporotic avian bone","volume":"310","author":"Knott","year":"1995","journal-title":"Biochem. J."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"S11","DOI":"10.1016\/S0020-1383(16)47003-8","article-title":"Bone mechanical properties and changes with osteoporosis","volume":"47","author":"Osterhoff","year":"2016","journal-title":"Injury"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"128","DOI":"10.1016\/j.commatsci.2011.01.021","article-title":"Micro-scale modelling of bovine cortical bone fracture: Analysis of crack propagation and microstructure using X-FEM","volume":"52","author":"Maligno","year":"2012","journal-title":"Comput. Mater. Sci."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"98","DOI":"10.1161\/01.ATV.6.1.98","article-title":"Induction of increased collagen and elastin biosynthesis in copper-deficient pig aorta","volume":"6","author":"Hill","year":"1986","journal-title":"Arteriosclerosis"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"149","DOI":"10.1017\/S0962728600030232","article-title":"Indicators of undernutrition in cattle","volume":"15","author":"Heath","year":"2006","journal-title":"Anim. Welf."},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Wehrle-Martinez, A., Dittmer, K., Back, P., Rogers, C., and Lawrence, K. (N. Z. Vet. J., 2022). Biochemical profile of heifers with spontaneous humeral fractures suggest that protein-calorie malnutrition could be an important factor in the pathogenesis of this disease, N. Z. Vet. J., in press.","DOI":"10.1080\/00480169.2022.2134226"},{"key":"ref_37","doi-asserted-by":"crossref","unstructured":"Counotte, G., Holzhauer, M., Carp-van Dijken, S., Muskens, J., and Van der Merwe, D. (2019). Levels of trace elements and potential toxic elements in bovine livers: A trend analysis from 2007 to 2018. PLoS ONE, 14.","DOI":"10.1371\/journal.pone.0214584"}],"container-title":["Biology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2079-7737\/11\/10\/1387\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T00:38:11Z","timestamp":1760143091000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2079-7737\/11\/10\/1387"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,9,23]]},"references-count":37,"journal-issue":{"issue":"10","published-online":{"date-parts":[[2022,10]]}},"alternative-id":["biology11101387"],"URL":"https:\/\/doi.org\/10.3390\/biology11101387","relation":{},"ISSN":["2079-7737"],"issn-type":[{"value":"2079-7737","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,9,23]]}}}