Project No. LAB03040 Start Date 1993 Oct 01 Term. Date 1999 Sep 30
Principal Invest. Chandler, J. E.
Title RELATIONSHIPS OF BULL SPERM CHROMOSOME CONTENT AND CONFIGURATION WITH SEMEN QUALITY & FERTILITY
Objectives To use genetic engineering techniques (PCR and FISH) to validate methods for separating bovine spermatozoa according to desired sex chromosome. To maintain seminal quality and fertility during the separation process. To determine from spermatozoal mitochondrial DNA (mtDNA), the proportions of embryonic mtDNA which is transmitted from the spermatozoa and how this may be affected by the use of sexed semen. To evaluate Dairy Herd Improvement Association records as a source of data for use in estimating bull fertility and how fertility may be influenced by the use of sexed semen.
Approach Sex separated sperm will be evaluated by PCR and FISH. Sperm DNA will be purified with enzyme digestion. PCR will be used to amplify sperm DNA by Taq polymerase. Amplified DNA will be electrophoresed on agarose gel stained with ethidium bromide. Gels will be photographed under UV light and analyzed for fluorescent intensity by image analysis. Sex separated sperm will be evaluated for viability and mitochondrial function using staining and videomicroscopy techniques. For quality compromised samples, motility will be stimulated with colostral whey, blood serum, and caffeine. Sire-daughter and daughter-dam mtDNA comparisons will be made to determine the extent of mtDNA inheritance from each parent using PCR techniques. Purebred and reciprocal crossbred progeny will be tested to quantify percentage of paternal mtDNA as affected by using sexed semen. Non-return data will be obtained from technician breeding receipts on cows that have records in the DHI database. Proportions of successful services from the DHI database will analyzed by weighted least squares methods. The weight used will be the number services per subclass. Means from the analysis will be used as estimates of bull fertility. Since DHIA data include birthday of calf and heifer calf identification for current lactation, LSNRATE will be used to monitor fertility of bulls whose semen has been sexed to ascertain how the separation process affects fertility.
Progress 1993/10 TO 1999/09
Objective: 1) To evaluate %Y chromosome bearing sperm (%YCBS) in separated and unseparated ejaculates with PCR; (2) to compare %YCBS data with %male calves and %male pigs per ejaculate; 3) to evaluate the collection frequency effect on %YCBS; and 4) to relate mitochondrial function to motility. Approach: A device (SEPDEVICE) was built to separate bull sperm by head size and ejaculates were separated. Sperm head area and head fluorescent intensity were used to indicate head size and measured with videomicroscopy. PCR with Y specific DNA evaluated y chromosome presence. We digitized and analyzed the fluorescent DNA PCR images for average intensity of PCR bands. Y band average intensity from individual ejaculates was compared with that of a 50% YCBS pool. %YCBS was estimated in multiple, unseparated ejaculates from ten sires. Breeding and calving data were merged and identified by cow, sire, herd and ejaculate. We evaluated the %male calves per ejaculate within sire basis. Farrowing data was evaluated for %male piglets per litter. We evaluated %YCBS on ejaculates from sexually rested bulls which were collected 6 times per week. Ejaculates from rested bulls collected twice per day, either weekly or every 21 days, were evaluated. Bulls were selected to maximize differences in mitochondrial populations. Boars were selected on maternal milk production EPD. We obtained semen from full sibling EPD groups from Landrace and Yorkshire breeds. Mitochondrial function was measured by methylene blue reduction rate(MeBRR)or ATP use rate. Motility and ATP concentrations were measured before and after incubation. Sperm numbers were assessed by spectrophotometry and motility was evaluated with a heated phase microscope. Semen was diluted to 1 hundred million cells/ml with a sodium citrate-glucose solution. Progress: Flourescent intensity of large sperm was less than bull WBCs (white blood cells). The small sperm flourescent intensity was greater than the WBCs. The interaction between bull and SEPDEVICE section for both sperm head area and fluorescent intensity of the same cells was significant. Head area and fluorescent intensity showed a size difference separation. Sperm quality decreased in both trials. Sires did not contribute to %YCBS variation. Both %male calves and piglets and PCR data showed %YCBS between ejaculates within sires varied beyond the sampling variance. Two of five ejaculates applied to the SEPDEVICE had high %YCBS and separated, but the others did not. In sexually rested bulls, ejaculate differences in %YCBS were significant in the first collection; remained large for the second; lessened for the third and fourth ejaculates and was least for the last two ejaculates. Minimal collection intervals reduced the ejaculate within bull variation in %YCBS. In bulls collected weekly, %YCBS changed sinusoidally with a period of 30 days. For bulls collected on a 21-day interval, after beginning high, %YCBS changed parabolically with a nadir at 40 days. Holstein mitochondrial efficiency differed from Brahman. Yorkshire mitochondrial efficiency was greater than Landrace. Correlation between sow milk production and son motility was moderate.

Impact: Semen from some bulls separated better than from others. By using collection frequency to optimize the %Y variation and identifying this variation by PCR, the use of ejaculate with naturally skewed sex ratios could enhance sex selection in cattle and swine production schemes. Differences in sperm mitochondrial efficiencies were discernible. Mitochondrial function was related to motility. Selection for motility, and thus fertility, should include an influence of the dam.
Publications 1993/10 TO 1999/09
1. Chandler, J. E., S.D. Degelos, A.M. Canal and J. B. Paul. 1999. A Technique for the Evaluation of Sperm Penetrating Ability and Quality of Bovine Semen Processed in an Extender made with Brackett-Oliphant Medium and Egg Yolk. Theriogenology. 51:1467-1476.

2. Chandler, J. E. , M.P. Wilson, A.M. Canal and H.C. Steinholt-Chenevert. 1999. Bovine Spermatozoal Head Size Variation And Evaluation of a Separation Technique Based on This Size. Theriogenology. 51:3155-3165.

3. J. E. Chandler . 2000. Cryoprotection of Sperm of Dairy Bulls. In: Advances in World Aquaculture, Vol. 7, Cryopreservation in Aquatic Species. T. Tiersch and P. Mazik, ed. World Aquacultural Society. pp 84-90.

4. Chandler, J. E., and A.M. Canal. 1999. The effect of collection frequency on percent Y-chromosome bearing spermatozoa in individual ejaculates from Holstein bulls. J. Dairy Sc. 82,Suppl 1:98-98.

5. Harrison, C.M., J. E. Chandler, and A.M. Canal. 1999. A spectophotometrical measure of methylene blue reduction for estimating mitochondrial function of bovine spermatozoa in correlation with motility. J. Dairy Sc. 82,Suppl 1:98-98.
Project No. LAB03307 Start Date  1998 Jan 01 Term. Date 2003 Dec 31
Principal Invest. Hay, G. M.
Title DAIRY CATTLE IMPROVEMENT THROUGH DHI
Objectives To provide support for a DHI program that will 1) provide Louisiana producers with data for improving efficiency of operations and 2) provide data for use in research and educational programs.
Approach The DHI program in Louisiana is run as a cooperative effort between the Department of Dairy Science, Dairy Extension, and the Louisiana DHIA. The state program is part of a national program involving the National DHIA and the USDA. A certified DHIA supervisor obtains individual cow milk samples and appropriate management data on a regular basis from participating herds. Results from milk analysis are compiled with farm data through a centralized computer system. A complete set of records is returned to the producer for use in decision making. Records are also made available to the Department of Dairy Science, Dairy Extension and the USDA for use in research and educational programs.
Progress 2002/01 TO 2002/12
The primary objective of the DHI program is to provide Louisiana producers with accurate records for improved decision making and efficiency of operation. Louisiana had 113 herds with 15,569 total cows participating in the DHI program at the end of 2002 giving an average herd size of 139 cows/herd. Although the number of herds participating in the DHI program declined by 23, herd size increased slightly for the year. Thirty-two producers are utilizing the pcdart program and five producers are using the newer pocket dairy program. Herds enrolled in the DHI program continue to produce over 4000 pounds more milk per cow per year than non-DHI herds.

Impact: The DHI program provides an accurate records program on both individual cow and herd bases. A dairy enterprise, like any other business, must have accurate records if it is to be managed in an efficient, business-like manner.
Publications 2002/01 TO 2002/12
No publications reported this period
Project No. LAB03312 Start Date 1998 Jan 01 Term. Date 2003 Dec 31
Principal Invest. Baham, A., Chandler, J. E.
Title A FULL SERVICE A.I. PROGRAM FOR IMPROVING GENETIC SUPERIORITY IN DAIRY AND BEEF LIVESTOCK
Objectives To provide scientific and operational support for a full A.I. program in dairy and beef cattle in Louisiana.
Approach The Dairy Improvement Center provides facilities and some scientific support for the A.I. program operated by Genex. Through the program in conjunction with Genex, a farmer-owned cooperative, and Cooperative Resources International, a wide selection of superior sires are available to breeders in Louisiana. The program also provides a range of services to the cattle breeders in the state. Dairy bulls are sampled annually in an effort to identify superior bulls for the program and increased emphasis is being placed on the custom collection programs for beef bulls.
Progress 2002/01 TO 2002/12
The Dairy Livestock Development Program Completed a successful year of service to the Louisiana dairy and beef industries. During this time period, AI program service and benefits were coordinated with activities of Genex/CRI Cooperative, Inc. The continuation of the 1996 Cooperative Agreement between Genex and the LSU Agricultural Center provides an operational basis for coordinating the AI activities carried out at the LSU Dairy Improvement Center as well as enhancing genetics for members and AI customers in Louisiana. Educational and research activities are coordinated with the Department of Dairy Science, the Louisiana Cooperative Extension Service, and other departments. Some of the accomplishments of 2002 include: 1)265,167 units of semen were collected from Genex and customer owned bulls with the majority of business in beef bulls; 2)the custom collection activity involved 24 beef breeds and 171 total bulls; 3)facility tours were carried out for 41 groups with participation vering from local livestock owners to international visitors; 4)herd analysis and mating programs for dairy producers and AI service to cattlemen in Louisiana has continued on a regular basis. In addition to continued promotion and participation in educational activities promoting AI, the AI program also coordinates and supports facilities and animals for research and teaching.

Impact: Artificial insemination is an important tool that provides livestock owners access to bulls proven to be of highest genetic superiority. The program provides breeders in Louisiana with a wide selection of superior sires along with a full service, custom collection program.
Publications 2002/01 TO 2002/12
No publications reported this period
Project No. LAB03548 Start Date 2001 Dec 01 Term. Date 2003 Jun 30
Principal Invest. Jenny, B. F.
Title DAIRY PLANT OPERATIONAL SUPPORT
Objectives To provide operational and technical support for all research, public service, and teaching activities related to dairy foods technology.
Approach A functional dairy processing plant and dairy store are maintained in the Dairy Science Building on the Louisiana State University campus. The dairy plant occupies about 7,700 square feet of space with facilities for receiving raw milk, fluid milk processing, and manufacture of ice cream, various cheeses, and other frozen and cultured dairy products. All raw milk required for research, teaching, and manufacture of frozen and cultured products is obtained from the LSU dairy farm. The dairy store occupies about 320 square feet of space and serves as a retail outlet for products produced in the dairy plant. Operation of the dairy plant and store is under the direction of a instructor with a appointment of 60% reseaarch and 40% teaching. A full time classified position assists with operation of the dairy store. The remaining labor force consists of part time student labor. This unit provides the technical and operational support for all research, public service, and teaching activities related to dairy foods technology.
Progress 2002/01 TO 2002/12
The primary objective of this project is to provide operational and technical support for all research, public service, and teaching activities related to dairy foods technology. Approximately 200 pounds of low and full fat cheeses and more than 40 gallons of various yogurts were produced in support of ongoing research projects. In addition, 1780 gallons of ice cream, 2570 pounds of cheddar cheese , and 120 gallons of eggnog were produced and marketed through the dairy store. Revenue generated is used to help maintain and operate the dairy plant. During the year, more than 3400 students, in grades ranging from elementary to college, toured the dairy plant.

Impact: A functional dairy processing facility is central to maintaining active research, public service, and teaching programs related to dairy foods technology.
Non-Tech. Summary A functional dairy processing facility is central to maintaining comprehensive research, public service, and teaching programs in dairy foods technology. The purpose of this project is to provide the technical and operationl support needed to conduct research, public service and teaching programs in dairy foods technology
Publications 2002/01 TO 2002/12
No publications reported this period
Project No. LAB93372 Start Date 1998 Nov 01 Term. Date  2002 Nov 01
Principal Invest. Williams, C. C.
Title REGULATION OF THE SOMATOTROPIC AXIS BY DIETARY PROTEIN IN GROWING RUMINANTS
Objectives 1. To determine the effects of dietary protein level and pattern of weaning on growth and anabolic hormone status in neonatal dairy calves; 2. To determine the effects of dietary protein level and rumen degradability on growth and hormonal status in weaned dairy calves.
Approach A study will be performed to determine whether weaning age (4 vs. 8 weeks) and calf starter crude protein level (16 vs. 22%)influence anabolic hormone levels and growth in pre-ruminant dairy calves. Feed intake, water intake, and health status of the animals will be recorded daily. Body weight gain and skeletal growth (wither height, hip height) will be measured. Blood samples will be collected and analyzed for growth hormone, insulin-like growth factor-I, insulin-like growth factor binding proteins, insulin, glucagon, and blood urea nitrogen. A second study will be conducted to investigate the effects of ruminally undegradable protein (RUP vs. no RUP) and dietary protein level (16 vs. 20%) on hormonal status and growth in weaned calves from 4 to 6 months of age. Measurements will include body weight gain and skeletal growth. Blood samples will be collected and analyzed for growth hormone, insulin-like growth factor-I, insulin-like growth factor binding proteins, insulin, glucagon, and blood urea nitrogen.
Progress 1998/11 TO 2002/11
Two trials investigated the effects of protein and amino acids on hormonal regulation in developing dairy calves. The first trial used 37 Holstein calves to determine effects of weaning age and dietary protein on growth and anabolic hormone status in neonatal dairy calves. Calves were assigned at birth to one of two weaning ages, either 4 or 8 weeks, and one of two starter crude (cp) protein levels, either 16 or 22% (as fed basis). Starter CP did not affect intake, body weight, or skeletal growth. Calves weaned at 8 weeks had greater body weights and heights than those weaned at 4 weeks. However, weaning calves at 4 weeks did not have negative effects on performance as these calves experienced no growth slumps after weaning. Concentrations of plamsa growth hormone (GH), IGF-I, insulin, glucagon, and glucose were not affected by starter CP. Plasma GH and glucose were greater in calves weaned at 8 weeks. Starter CP did not affect hormone or metabolite concentrations, but age related changes in these metabolic parameters were identified. The second trial used 24 Holstein heifer calves to evaluate the efficacy of amino acids as GH secretagogues in neonatal dairy calves and to monitor changes in these responses as calves undergo the transition to becoming fully functioning ruminant animals. Calves were assigned to one of 4 treatments including physiological saline, arginine, aspartic acid, and ornithine. At 1 and 3 months of age, amino acid challenges were conducted. There was an acute response in plasma GH to aspartic acid infusion in calves at both ages. Plasma GH decreased with age. At 1 and 3 months, plasma insulin concentrations increased in response to arginine and ornithine followed by a decrease in glucose concentrations.

Impact: Despite slight increases in growth in calves weaned at 8 weeks, dairy calves can be successfully weaned at 4 weeks of age. Additionally, increasing the protein level in calf starter to 22% does not improve growth or influence metabolism. Therefore, weaning at an earlier age and not purchasing higher protein starter feeds may provide an economic benefit to dairy producers. Supplemental amino acids which may improve growth through increasing anabolic hormone concentrations warrent further investigation.
Publications 1998/11 TO 2002/11
No publications reported this period
Project No. LAB93416 Start Date 1999 Oct 01 Term. Date 2002 Sep 30
Principal Invest. Williams, C. C.
Title MANAGEMENT SYSTEMS FOR IMPROVED DECISION MAKING AND PROFITABILITY OF DAIRY HERDS
Objectives 1. Develop and integrate decision support systems to promote efficient, environmentally sound, and economically viable management systems for dairy youngstock. 2 Develop and integrate decision support systems to optimize productive and economic efficiencies for lactating and dry cow management systems.
Approach The effects of dietary protein and ruminally undegradable protein on growth of weaned dairy heifers will be studied. Holstein heifer calves, approximately 4 months of age, will be assigned to one of four dietary treatments consisting of two grain supplements containing either 16 or 20% crude protein with or without a source of rumen escape protein. Body weight and height will be monitored. Anabolic hormone (growth hormone, IGF-I, insulin) status will also be assessed in order to determine whether increasing levels of protein and/or post-ruminal supply of amino acids will improve growth through increasing the concentrations of these hormones.
Progress 1999/10 TO 2002/09
Two projects investigated nutritional management regimens for improving performance in neonatal dairy calves. The first project used 32 Holstein calves to determine effects of vitamin E and/or lasalocid on growth and immune responses in calves challenged with Eimeria bovis. Including lasalocid in milk replacer and calf starter provided adequate protection against clinical signs of coccidial infections in neonatal calves. The calves remained healthy and did not experience depressed starter intake and decreased body weight gains as did the calves supplemented with vitamin E alone. While lasalocid was effective in decreasing severity of coccidiosis, vitamin E alone was not. Incidence of scours increased in calves fed vitamin E. The second project used neonatal Holstein and Jersey calves to investigate effects of feeding milk replacer once or twice daily on growth and glucose metabolism. Feeding calves milk replacer once or twice daily did not affect starter intake or growth or adversely affect glucose metabolism in Holstein or Jersey calves. Overall, milk replacer feeding frequency did not affect plasma NEFA, glucose, insulin, or glucagon concentrations. Data from the frequently sampled glucose tolerance tests confirmed that feeding calves milk replacer once daily did not increase insulin resistance or negatively affect glucose metabolism in Holstein or Jersey calves. These data provide evidence of metabolic differences between Holstein and Jersey calves.

Impact: Including lasalocid in milk replacer and calf starter will benefit neonatal dairy calves by providing protection from coccidial infections. Addition of supplemental vitamin E to neonatal calf diets does not appear to improve their ability to resist coccidiosis. While vitamin E has been shown to improve the immune system in other aspects of dairy management, it did not benefit neonatal dairy calves in this study. Since feeding milk replacer once daily did not adversely affect growth or glucose metabolism in neonatal dairy heifers, this may provide an economic benefit to dairy producers in terms of decreased cost of labor. However, young dairy calves should be monitored more than once daily as part of a heifer management program.
Publications 1999/10 TO 2002/09
Stanley, C.C., C.C. Williams, B.F. Jenny, J.M. Fernandez, H.G. Bateman, II, W.A. Nipper, J.C. Lovejoy, D.T. Gantt, and G.E. Goodier. 2002. Effects of feeding milk replacer once versus twice daily on glucose metabolism in Holstein and Jersey calves. J. Dairy Sci. 85:2335-2343.
Project No. LAB93441 Start Date 1999 Oct 01 Term. Date 2004 Sep 30
Principal Invest. Chandler, J. E.
Title SEX CHROMOSOME VARIATION AS RELATED TO QUALITY, FREEZABILITY, FERTILITY AND SEX RATIO IN FARM SPECIES SPERMATOZOA
Objectives 1. To use techniques such as image analysis or polymerase chain reaction (PCR) to select ejaculates of variable primary sex ratio both within and among sires to be used in the manipulation of sex rations in producers' calf and farrowing crops. 2. To improve the efficacy of a patened semen separation appratus both as to sperm number yield and sperm quality. 3. To determine how spermatozoal separation technologies affect semen quality, particularly spermatozoal mitochondrial function, i.e, motility. 4. To determine relationship between sperm mitochondrial function, as measured by methylene blue reduction, mitochondrial DNA fingerprint, parental energy use and parental mitochondria DNA fingerprint.
Approach OBJECTIVE 1: Boar ejaculates will be collected from boars, extended and an aliquot will be taken while the remainder of each ejaculate will be used for AI. Breeding and farrowing records will be compiled for statistical analysis. Sperm DNA will be extracted and amplified in a duplex PCR with unique sequences from the X and Y crhomosomes. Digital images of agarose gels with ethidium bromide stained PCR products will be obtained for image analyses. The intensity density of each band will be corrected with an internal standard and then be compared to an external standard. White blood cells (WBC) DNA obtained from boars and sows will be used as the external standard. The WBCs from each sex will be pooled for an X: Y chromosome gradient. The WBC-DNA will be extracted, amplified, and electrophoresed alongside the samples. The WBC PCR product band will be compared with individual samples on a within gel basis. Primary sex ratio data from PCR analysis will be compared with secondary sex ratio data obtained from farrowing records. Experiment 2: Semen will be collected from boars and individual ejaculates screened using the procedure described in Experiment 1. Semen will be extended and frozen, pending PCR screening results, used for AI. Ejaculates that differ from a 1:1 sex ratio will be used in AI. Resulting piglet gender data will be compiled and compared with PCR data to determine the predictability of the screening process for secondary sex ratios. The PCR screening protocol will be adapted to bovine semen. OBJECTIVE 2. To improve the separation efficiency, experiments will be conducted with the SEPDEVICE using bull semen with known %Y-chromosome bearing spermatozoa. Ejaculates that contain a high percentage of large, or X-chromosome bearing, sperm will be manimpulated to attempt to decrease the incidence of clogging. The dimensions of the SEPDEVICE will be changed to accommodate a larger volume of semen while maintaining the original design principles. Semen handling methods will be adopted to sustain the quality of the spermatozoa as they are being processed through the SEPDEVICE. Acrosomal integrity and motility of the spermatozoa will be monitored to evaluate the impact of the modification of the design or application of the SEPDEVICE on seminal quality. Specific PCR with image analysis will be used to evaluate separation effectiveness. OBJECTIVE 3: Fresh bovine ejaculates will be used for the methylene blue testing and motility. Blood samples will be drawn from the dams of these bulls. White blood cells will be harvested and cultured to increase cell numbers. Cell viability will be evaluated by trypan blue. Methylene blue in a glucose containing buffer will be added to the culture tubes. Methylene blue reduction rates will be calculated similar to that reported for spermatozoa. OBJECTIVE 4: Spermatozoal mitochondrial DNA fingerprints will be used to relate mitochondrial fingerprints from the bull and dams. Spermatozoa will be decapitated and the head and tails will be separated with Percoll. The mtDNA will then be extracted and random PCR primers will be used to produce profiles comparison between related individuals.
Progress 2002/01 TO 2002/12
OBJECTIVE: Mitochondrial DNA (mtDNA) was extracted from semen to compare the relationship of point mutations to motility and ATP use. ACCOMPLISHMENTS: Ejaculates (n=16) were obtained via an artificial vagina from two Holstein and two Brahman bulls. Semen was processed and stored in 0.5 ml French straws. Polymerase chain reaction was used to amplify a 955 bp sequence of the NADH1complex I gene. The sequence was divided into two segments: segment 1 (SEG1) (range 3131 to 3588bp); segment 2 (SEG2) (range 3567 to 4055bp). The DNA sequence of each segment was compared to the appropriate segment of mtDNA sequence of the Bos taurus heart. The occurrence of point mutations was expressed as percent of disagreements (%DISAGREE) with heart mtDNA and percent of ejaculates with more than 1 mutation at the same position (%GT1MUT@P) in the DNA sequence. Data were analyzed by least squares and correlation methods. In the first study in the Holstein ejaculates, SEG1 had 33.0 %DISAGREE and 60%GT1MUT@P while SEG2 had 26.6 %DISAGREE and 83.9%GT1MUT@P. Sequence analysis is currently being done on the Brahman ejaculates. The second experiment was done to detect the effects of mtDNA point mutations on sperm motility and ATP use in Holstein and Brahman bulls. Percent progressive motility was measured immediately post thaw (MOT0) and again after a 3h-37C incubation (MOT3). ATP concentration was measured by the luciferin/luciferase reaction and detected in a scintillation counter before (PMOLES0) and after the incubation (PMOLES3). The average motility of the thawed semen across all ejaculates was 34.4 %. Strong positive associations existed between motility and ATP concentrations. Correlation between MOT0 and MOT3, PMOLES3, %DISAGREE in SEG2 and %GT1MUT@P in SEG2 were significant. Significant negative correlations existed between the metabolic parameters (MOT0 and PMOLES3) and the SEG2 sequence information (%DISAGREE and %GT1MUT@P). These correlations were all greater than 70%. SIGNIFICANCE: Based on these results, there could be a relationship between the number of point mutations in mitochondrial DNA and sperm motility and metabolism. As the number of mutations increased, sperm motility and metabolism decreased. Since it is believed that the mitochondrial population in mammalian species is largely derived from the maternal cytoplasm, this could imply that dam information should be included in pedigree designs of future bulls. This would help maintain or improve the bull's fertility.

Impact: Currently, when the pedigree of a new dairy bull is constructed, information from the dam is not directly included. Since sperm motility is a function of the mitochondria and is involved in the fertilization process, and if mitochrondria are derived from the mother, it would seem necessary to include her information in the pedigree in order to influence her son's sperm motility and thus his fertility.
Non-Tech. Summary Offspring gender selection in farm animals will enable meat and milk producers to increase production efficiencies by reducing the number of the less or non-productive sex. The relationship between mitochondrial function and sperm motility is needed to better evaluate bull fertility. This project evaluates a technique for picking ejaculates that are skewed from the 50:50 X to Y chromosome bearing sperm. Understanding the sperm motility can be used in evaluating bull fertility. The project will cover the futher development of a patented device to augment the naturally occurring skew.
Publications 2002/01 TO 2002/12
R. Paul Lang, Kenneth L. Riley, John E. Chandler and Terrence R. Tiersch. 2003. The use of dairy protocols for sperm cryopreservation of Blue Catfish, Ictalurus fucatus. Journal of the World Aquaculture Society. 34, (1):66-75.
Project No. LAB93473 Start Date 2000 Jul 01 Term. Date  2004 Jun 30
Principal Invest. Bateman, H. G.
Title USING AMINO ACIDS TO IMPROVE RUMINAL FERMENTATION AND MILK PRODUCTION BY LACTATING DAIRY COWS
Objectives 1. To determine if providing amino acids to the ruminal environment will stimulate ruminal fermentation in vitro. 2. To determine the effects of feeding amino acids to lactating dairy cows on ruminal fermentation and duodenal flow of dry matter, organic matter, N, fiber, and fatty acids. 3. To determine the effects of feeding amino acids to dairy cows on milk production, milk composition, and production of milk components.
Approach A three phase approach will be used. In phase 1, ruminal fermentation in continuous culture will be used to investigate the effects of methionine and lysine on ruminal fermentation. Phase 1 will also be used to determine levels of methionine and lysine that are expected to result in changes in ruminal fermentation when fed to lactating dairy cows. Phase 2 will determine the effects of feeding methionine and lysine on duodenal flow of nutrients. In this phase, lactating dairy cows will be surgically modified and fitted with cannula's in the rumen and duodenum. These cows will be fed diets that contain differing levels of free methionine and lysine (levels determined from phase 1) and characteristics of ruminal fermentation and duodenal flow of nutrients measured. In phase 3, 40 lactating dairy cows will be fed diets differing in levels of free methionine and lysine to measure the effects of these amino acids on milk production and milk component production.
Progress 2002/01 TO 2002/12
Projects have been completed that characterize the impact of supplemental Zn on rumen fermentation and degradation of urea. Information collected indicates that supplemental Zn does not impair ruminal degradation of urea when fed with high quality diets. This is in contrast to other data that indicates that supplemental Zn does impair ruminal degradation of urea when fed in low quality diets. Sample collection is complete for trials investigating the impact of supplemental methionine and lysine on fermentation in vitro. Laboratory analysis of the collected samples is currently underway. A cow feeding trial using supplemental methionine and lysine was also conducted. The data from this trial are being statistically analyzed. Completion of the analysis for the last two trials (amino acid supplementation trials) will provide information that can be used to determine if supplementation of these two amino acids will provide a significant benefit to ruminal fermentation that can be exploited by the host animal.

Impact: Feed costs continue to be the largest single expense for dairy producers after labor. This trial will provide information that will allow producers to make better-informed decisions when choosing protein supplements for dairy cattle.
Non-Tech. Summary Feed costs account for approximately 40 percent of the total cost of milk production by a dairy farm. Feeding free amino acids may improve fermentation in the rumen and allow for more accurate feeding of dairy cows. This trial is designed to gather data that will allow a dairy producer to determine if feeding free methionine and lysine will improve ruminal fermentaion and increase milk production or decrease their feed cost.
Publications 2002/01 TO 2002/12
Bateman, II, H. G., C. C. Williams, and Y. H. Chung. 2002. Effects of supplemental Zinc in high quality diets on ruminal fermentation and degradation of urea in vitro and in vivo. Prof. Anim. Sci. 18:363-367.
Project No. LAB93560 Start Date 2002 Jan 01 Term. Date 2004 Dec 31
Principal Invest. Bateman, H. G.
Title NUTRITIONAL MANAGEMENT STRATEGIES FOR OPTIMAL ENERGY METABOLISM IN TRANSITION DAIRY COWS
Objectives 1. To estimate the influence of prepartum energy density on metabolic markers of energy metabolism in transition dairy cows. 2. To estimate the influence of supplemental gluconeogenic precursors in the diet (Ca salts of propionic acid) on metaboilic markers of energy metabolism in transition dairy cow. 3. To determine if urinary measures such as pH can be used to estimate incidence of ketosis in transition dairy cows.
Approach Forty Holstein dairy cows will be blocked by anticipated parturition date and parity and used in a randomized block designed experiment to investigate the feasibility of using urinary measurements to predict incidence of ketosis in transition dairy cows. Treatments will be one of four diets that are arranged as a 2 x 2 factorial. Factors are prepartum dietary energy concentration (normal for non-lactating dairy cows and high) and addition of a gluconeogenic precursor (Ca salts of propionic acid). As offered feed intakes will be measured daily using electronic feeding gates. After parturition, energy concentration of the diet will be increased for all cows to a level comparable to that of a normal diet for support of lactation. Diets will be sampled daily. Individual feed ingredients and refused feed will be sampled weekly. Samples of diets, ingredients, and feed refusals will be dried and ground and analyzed for OM, NDF, ADF, CP, and EE. Milk production will be recorded and sampled at each milking. Milk samples will be pooled bi-weekly and analyzed for fat, protein, and somatic cell content. Blood samples will be taken 3x/week throughout the study. Plasma will be isolated and stored frozen. Stored plasma will be composited by week relative to actual parturition date and analyzed for ketones, glucose, total Ca, plasma urea N, nonesterified fatty acids, insulin and thyroid hormone concentrations. Urine will be sampled 3x/ wk throughout the trial and will be immediately assessed for pH and ketone levels. The urine samples will then be stored for later lab analysis of ketones. Body weight of cows will be recorded each week during the study. The same forty cows from the experiment outlined above will be used in a similar experiment to determine the effects of providing gluconeogenic precursors in the diet on energy metabolism in transition dairy cows. During weeks -1 relative to anticipated parturition date and +1 relative to actual parturition date FSIVGTT will be performed. Data from the FSIVGTT will be used to assess glucose effectiveness, insulin sensitivity and the acute insulin response relative to glucose administration using the minimal modeling computer program. The FSIVGTT consists of an infusion of glucose (500mg/kg BW) followed 20 min later by infusing bovine insulin (0.03 IU/KG BW) with repeated sampling of blood for 180 min. Data from the FSIVGTT will be processed using the minimal modeling computer program and values for glucose effectiveness, insulin sensitivity, and the acute insulin response relative to glucose administration obtained.
Progress 2002/01 TO 2002/12
Animal data collection from two lactating cow trials has been completed. Trial one investigated the use of urine pH measurement for prediction of the incidence of ketosis. Cows were followed from 3 weeks prepartum until 3 weeks postpartum. Urine pH and ketone levels were measured 3x/week and will be used to formulate predictive models. Trial two investigated the impact of calcium propionate on glucose metabolism around parturition. The frequently sampled intravenous glucose tolerance test (FSIVGTT) will be combined with the minimal model computer analysis to investigate the impact of prepartum dietary energy level (105 or 145% of NRC requirements, 2001) with or without addition of 113.5 g Ca-propionate /d on the metabolic response to a glucose and insulin challenge. The FSIVGTT were conducted at 1 week prior to anticipated parturition date and 1 week post-partum. Laboratory analysis of collected samples is underway and data will be statistically analyzed.

Impact: This project will provide information that dairy producers can use to better manage and prevent the metabolic disorder ketosis. We expect that the project will provide a method for rapid evaluation of cows around the transition period to determine if they are ketotic and will also provide information about the underlying metabolism of those cows that will improve treatment regimes.
Non-Tech. Summary Ketosis is a metabolic disorder that affects approximately 7 to 32 percent of dairy cows in the US. This project is designed to determine if easily measured cow parameters such as urine pH can be used to predict the incidence of ketosis. A secondary objective of this project is to evaluate a commercial supplement for its ability to stimulate energy metabolism in transition dairy cows.
Publications 2002/01 TO 2002/12
No publications reported this period
Project No. LAB93585 Start Date 2002 Jun 01 Term. Date 2006 Jun 01
Principal Invest. Aryana, K. J.
Title CHARACTERISTICS OF SOME FERMENTED DAIRY FOODS AS INFLUENCED BY THE INCORPORATION OF FOLIC ACID AND CHITOSAN
Objectives 1. To study the concentration and stage of addition effects of folic acid in the manufacture of fat free sugar free plain set yogurt and flavored yogurts. 2. To determine the concentration effects of three types of chitosan on the characteristics of full fat and low fat Cheddar cheeses.
Approach Yogurt fortification with folic acid. Fat free sugar free plain set yogurt will be manufactured at the LSU Dairy Processing Plant, Department of Dairy Science, LSU. The flavor used for the manufacture of flavored yogurts will be strawberry and lemon. Plain and flavored yogurts will be manufactured with 0, 25, 50, 75 and 100 percent of the RDA for folic acid in a single cup of yogurt. Folic acid will be added before and after pasteurization of the mix. Protein, fat, moisture and ash will be determined one week after production. Folic acid concentration at different stages of processing will be determined at weeks 1 and 5. Apparent viscosity, pH, titratable acidity, syneresis, protein / peptide profiling, colorimetry, sensory scoring and descriptive, coliforms, yeasts and molds, standard plate counts will be studied at 1,3 and 5 weeks after production. Full fat and low fat Cheddar cheeses manufactured with chitosan. Full fat and low fat Cheddar cheeses will be manufactured at the Dairy Processing Plant, Department of Dairy Science, LSU. Three different forms of chitosan viz. regular, dietary supplement grade and 90% high density, will be used. Chitosans will be incorporated at 0, 0.5, 1 and 1.5 percent w/w in cheese or w/w fat in cheese, to be determined based on organoleptic evaluation during preliminary experiments. Protein, fat, moisture and ash content in cheese will be determined one week after the manufacture of cheese. Texture, protein profiling, colorimetry, pH, sensory scoring and descriptive, coliforms, yeast and molds and standard plate counts will be studied at 1, 8, 16, 24 weeks after cheese manufacture.
Non-Tech. Summary Consumer demand exists for good tasting foods containing few calories and more health benefits. Folic acid is known to have several health benefits related to neural tube defects, coronary heart disease, colorectal and cervical cancers. Chitosan is a fat absorbing fiber, which could make it possible to enjoy the good taste and texture advantages of fat in the foods without the fat calories. Whether or not folic acid and chitosan alter the characteristics of dairy foods is not known.
Project No. LAB93593 Start Date 2002 Oct 01 Term. Date 2007 Sep 30
Principal Invest. Williams, C. C.
Title MANAGEMENT SYSTEMS TO IMPROVE THE ECONOMIC AND ENVIRONMENTAL SUSTAINABILITY OF DAIRY ENTERPRISES
Objectives 1. To evaluate and develop nutrient utilization schemes for dairy herd replacement heifers with implications to economic efficiencies and environmental impacts. 2. To develop strategies and systems to optimize nutrient utilization, economic and financial returns, and environmental goals for management of dry, pregnant, and lactating dairy cows.
Approach Nutritional and management strategies for utilization of energy, protein, amino acids, and selected macrominerals that meet requirements of young calves and growing heifers for optimal growth and reduced excretion into the environment will be studied. Studies will be used to validate and/or refine the current NRC (2001) requirements. The project participants also propose to elucidate biological processes that differentiate the ability of individual calves to resist or succumb to high morbidity beyond that traditionally associated with passive immunity. The portion of this project specifically being conducted by this researcher will determine effects of dietary protein level and rumen undegradable protein on growth of weaned dairy heifers will be studied. Holstein heifers, approximately 4 months of age, will be assigned to one of four dietary treatments consisting of two grain supplements containing either 16 or 20% crude protein with or without a source of rumen escape protein. Body weight and height will be monitored. Concentrations of growth hormone, insulin-like growth factor-1, and insulin will be measured to determine whether increasing levels of protein and/or post ruminal supply of amino acids will improve growth through increasing the concentrations of these anabolic hormones.
Non-Tech. Summary A. Little technology has been employed to dairy heifer replacement management in a holistic systems approach to reduce costs, increase feed efficiency, and reduce nutrient losses to the environment. B. The changes and trends in the US dairy industry have led to the need for improved decision support systems which will enable better farm management. This project examines nutrient utilization in dairy heifers and mature dairy cows for optimum performance that is economically efficient and environmentally sound.
Project No. LAB93598 Start Date 2002 Oct 01 Term. Date 2007 Sep 30
Principal Invest. Bateman, H. G.
Title METABOLIC RELATIONSHIPS IN SUPPLY OF NUTRIENTS FOR LACTATING COWS
Objectives 1. To quantitatively evaluate the chemical and physical properties of protein and energy sources which determine the availability of nutrients critical to milk protein secretion in lactating dairy cows. 2. To measure the metabolic interactions among amino acids, lipids and glucose which affect synthesis of milk components.
Approach Experiments will be conducted with continuous culture fermentors to evaluate protein supplementation schemes on microbial fermentation of carbohydrate and microbial protein synthesis. Additional experiments will be conducted with intact and cannulated cattle to determine if results obtained in culture can be extrapolated to use with live animals.
Non-Tech. Summary Protein is a costly nutrient that has enormous potential for adverse impact on the environment when overfed. Improving estimates of the minimal protein requirements for maximal microbial fermentation in the rumen should allow for greater milk production efficiency while minimizing the potential for this negative impact on the environment. The purpose of this trial is to optimize protein supplementation strategies to provide for optimal microbial fermentation in the rumen of lactating cows.