Utah State University DigitalCommons@USU All Graduate Theses and Dissertations Graduate Studies 5-1991 Changes in the Vulvular and Vestibular Tissue of the Bovine During the Estrous Cycle as Determined by the use of Near Infrared Interactance Robert Alan Kunzler Utah State University Follow this and additional works at: https://digitalcommons.usu.edu/etd Part of the Animal Sciences Commons Recommended Citation Kunzler, Robert Alan, "Changes in the Vulvular and Vestibular Tissue of the Bovine During the Estrous Cycle as Determined by the use of Near Infrared Interactance" (1991). All Graduate Theses and Dissertations. 4222. https://digitalcommons.usu.edu/etd/4222 This Thesis is brought to you for free and open access by the Graduate Studies at DigitalCommons@USU. It has been accepted for inclusion in All Graduate Theses and Dissertations by an authorized administrator of DigitalCommons@USU. For more information, please contact [email protected]. CHANGES IN THE VU LVULAR AND VESTIBULAR TISSUE OF THE BOVINE DURING THE ESTROUS CYCLE AS DETERMINED BY THE USE OF NEAR INFRARED INTERACTANCE by Robert Alan Kunzler A the sis submitted in partial fulfillment o f the requirements for the degree of MASTER OF SCIENCE in Dairy Science a pproved: Commi ttee Me mber Dean of Graduate Studies UTAH STATE UNIVERSITY Logan, Utah 1991 ACKNOWLEDGMENTS ii This research project was initiated in the spring of 1990, under the direction of Dr. David P. Marcinkowski. I would like to express my sincere appreciation to Dr . Marcinkowski for his patience, encouragement, and technical help during the project and in analyzing the data. I would also like to thank the other members of my graduate committee for their time, wisdom , and expertise in their respective fields. Thanks to Dr . Ellis for his suggestions, Dr. Bunch for help in advising on classes that would help me to understand my project better, and Dr. Clark for his technical expertise with the use of the Near infrared reflectance spectrophotometer and in analyzing the data once it had been collected. Others who assisted in the collection of the spectra included Cesear Acosta, Hassan Osmond, Beth Howery and Virginia Palimano. In addition, Javhar Raj analyzed the blood progesterone for the project. I owe my sincere thanks to my parents, Mark and Doris Kunzler, and to my family for the love and support they have given me in school. Finally, I extend my appreciation to my wife, Wendee , for her patience, love, and support during the accomplishment of this goal. Robert Alan Kunzler TABLE OF CONTENTS i ii Page ACKNOWLEDGMENTS ............... ............... .. ... ii LIST OF TABLES. i v LIST OF FIGURES. • . v ABSTRACT ..•. .. .•. ................. .. .. ..... .... .. vi STATEMENT OF THE PROBLEM .. .. .. ....... ........ .. ....... 1 I ntroduction.................. .. .. ... .. ....... 1 Problem Statement ..... ...... • . .. ... .. .... .. .. .. .. 5 Purpose and Objecti ves .... .... ................ ..... 6 LITERATURE REVIEW. 7 The Estrous Cycle . 7 Female Reproductive Organs . ... .. .. ... .. .. ... ... .. 10 Histological Changes in the Vagina during the Estrous Cycle . 13 Changes in the Vulvular Mesenchyme during the Estrous Cycle. 14 v aginal Mucus. 18 Estrous Detection Methods .............. .. .. .... .. .. 20 Near Infrared Spectra ... .. ...• . •.. ....... ... .. 28 Near Infrared Spectrophotometer Instrumenta t i on . .. .. 32 MATERIALS AND METHODS ......... ......... •. .. .. .. ... ..... 36 Introduction .......... .. .... .......... .. ... ...... 36 Collection of the Spectra ..... ....... .... ... ... ..... 40 Editi ng of Observations ........ ...... ... ..... .... 41 Treatment of the Spectra ... ... ............... .. 45 RESULTS AND DISCUSSION ............. • ...... .. .......... 48 Sample Identification .. ......................... .... 48 Interactance Spectral Modification .... ........ .. .. 53 Regression Analysis.. 55 C~ang7s.in the Spec~ra: Estrus vs . Nonestrus . .... .. 59 D~scr~m~nate Analys~s. 68 CO NCLUSIONS. 7 6 REFERENCES ... ............ ..... ......... .. .. ... ....... .. 79 APPENDIX ................. ... ....... ...... ... .. ... 86 LIST OF TABLES iv Table Page 1 Reasons for the exclusion of cows from the final data set .... ..... ....................... .. ... 43 2 Reasons for the exclusion of estrous periods from the final data set ..... .. ... ... ... ... .. ....... 46 3 Summary of Production .and Reproduction of cows ... 49 4 Results of the Partial Least Squares Linear regression of the vestibule and vulva (N=486) . Va riable day ... ..... .. ... ... ..... ... ... ..... .. 57 5 Results of the Partial Least Squares Linear regression of the vestibule and vulva (N=486 ). Variable heat ..... .... .. ... .. .............. .... 57 6 Results of the discriminant analysis of the vestibule first derevative (Error rate 26%) ...... 70 7 Frequency distribution of the misclassified days .71 8 Results of the discriminant analysis of the vestibule in the second derivative (Error rate 37%) .......... ... .................. .. 73 9 Results of the discriminant analysis of the vulva in· first derivative (Error rate 34%) ....... 74 10 Results of the discrimi nant analysis of the vulva in second derivative (Error rate 34 %) ... ... 74 v LIST OF FIGURES Figure Page 1 Changes in the cellular water content ....... ... 16 2 Near Infrared Spectra of water (log1/R) ........ 30 3 Spectra collected from 1100 to 2500 ........ ... 54 4 Outside spectra (log 1/I) .................. ... 60 5 Inside spectra, (log 1/I) ......... ........ .. 61 6 Vestibular spectra first derivative . .. ... .. ... 63 7 Vestibular spectra second derivative ........... 66 8 Vulvular spectra first derivative ..... ........ 67 9 Vulvular spectra second derivative ........ .... 69 vi ABSTRACT CHANGES IN THE VULVULAR AND VESTIBULAR TISSUE OF THE BOVINE DURING THE ESTROUS CYCLE AS DETERMINED BY THE USE OF NEAR INFRARED INTERACTANCE by Robert Alan Kunzler, Master of Science Utah State University, 1991 Major Professor: Dr. David P. Marcinkowski Department: Animal, Dairy, and Veterinary Sciences Near infrared reflectance spectroscopy is routinely used for the analysis of quality components of feedstuffs. Near infrared spectrophotometers, coupled with a fiber optic probe, could enable direct measurements of the live animal. This study was conducted to characterize changes in the vulva and vestibule during the bovine estrous cycle using near infrared (NI) spectroscopy. Sixteen cycling Holstein cows were observed for estrus twice daily from 40 days postpartum for three estrous cycles or until conception was verified. In addition, weekly rectal palpations, cowside milk progesterone tests, and tailhead chalk were used to aid in estrous detection. Near infrared spectra of both the vulva and the vestibule were collected daily (Model 6500 with 1.83 m fiber optic probe, NIR Systems Inc., 1100 to 2500 nm). Twenty-four estrous periods were confirmed. Vul vular and vestibular Vil spectra of estrous and nonestrous cows were compared. Spectral differences occurred in three regions at 1700 nm, 1790 nm, and 1880 nm (P < .OS) . These regions are possibly associated with changes in carbohydrate, protein, and water content. Results indicate that direct NI analysis of the live cow is possible and changes in the vulvular and vestibular tissue during the estrous cycle can be detected. However, accurate estrous detection using NI interactance is not practical at the present time because of individual cow variability . (93 pages) STATEMENT OF THE PROBLEM Introduction Milk production and reproductive performance play major roles in determining the profitability of a dairy herd. Milk production is dependent on reproduction because lactation is induced by pregnancy and calving. Milk production also depends on the cow's genetics and the environment she is exposed to . Therefore, a knowledge of the relationship between milk production and reproduction is important for profitability of the dairy. Shanks et al. (61) associated 40% of the dairyman's direct health costs with r eproduction. These costs were divided between reproductive and insemination costs. It has been estimated that 20% to 25% of all dairy cows are culled because of poor reproductive performance ( 11) . Expenses associated with reproduction increase dramatically when the cost of replacing these culled cows is considered. In 1981 it was estimated that United States dairymen lost $960 million because of poor reproductive performance (31). Artificial insemination (AI) is a valuable tool for the genetic improvement of animals. In Utah, 80% of the dairy cattle are artificially inseminated (43). AI allows the dairyman to improve his herd through the genetic improvement of the cows, to control venereal diseases, and to increase safety by eliminating the use of dangerous bulls on the dairy . The use of artificial insemination has made accurate estrous detection a key to the reproductive performance of a dairy 2 herd. Field studies have shown that only 52% of estrous periods are detected ( 4 0, 64) . The trend toward larger herds, and more efficient feeding arrangements minimizes contact between the cow and the herdsman. Less contact makes it more difficult to identify cows in estrus. Other problems dairymen encounter in the detection of estrus include extremes in temperature, high production, confinement housing, and nutritional limitations, causing an energy imbalance, all of which reduce the expression of estrus (63, 66, 68). Visual aids have been developed to help the dairyman dete ct more estrous