Due to necessary storage of hatching eggs in commercial hatcheries, the embryo is being regressed. End of lay broiler breeder eggs have the poorest hatchability and are most affected by pre incubation storage. Warming these eggs is the only way the embryo can develop. The objective of this study was to study the effects of daily warming of end of lay broiler breeder hatching eggs during the storage period on embryo mortality and hatchability. Six trials were conducted, three trials warmed for three of the four day storage period and three trials warmed for five of the six days storage period. There were six warming treatments for each trial; 0, 30, 60, 90, 120, and 150 minutes daily. All eggs were from Hubbard Classic broiler breeders 50-58 weeks of age. Hatch time was recorded for trials 3 and 6. Two hundred and eighty eight randomly selected males were used in trial 5 to study the effects of daily warming on chick growth. They were fed a starter broiler diet and grown for 13 days in a Petersime starter battery. In all hatchability trials percent fertility, percent fertile hatchability, percent total hatchability, percent early dead, percent mid-dead, percent late dead, percent pips, and percent total embryonic mortality was not affected (P>0.05) by any of the daily warming treatments. Hatch time and chick growth were not affected by any of the daily warming treatments. Eggs could be warmed for as much as 150 minutes daily during storage without affecting hatchability which contradicts to management procedures. These results suggest that it is unnecessary for refrigerated trucks to transport eggs from the farm to the hatchery.

A study was conducted to determine effects of differing fat and protein levels in commercial milk replacers on the growth performance and rumen development of neonatal Holstein calves. Fifty-three calves (23 male; 30 female) were assigned to one of three dietary treatments which included 3 milk replacers (20/20; 25/15; 28/20) fed once per day at 10% of birth weight through weaning. Calves were offered a 20% CP calf starter and water free choice beginning on day 5. All calves were weaned on day 42. Body weights were measured at birth and weekly through weaning. Additionally, hip height, wither height, and body length were measured weekly. Feed intake and fecal scores were recorded twice daily. Beginning on day 7 and continuing weekly through weaning, blood samples were collected prior to morning feeding for analysis of IGF-I and on days 14, 28, 42, and 56 for PUN and â-hydroxybutyrate concentrations. On days 28 and 56 rumen fluid was collected for analysis of pH and VFA to evaluate possible differences in rumen development. Calves fed 28/20 milk replacer had greater (P < 0.05) daily starter intake, body weights, PUN, and â-hydroxybutyrate concentrations when compared to calves fed 20/20. There was also a treatment by week interaction (P < 0.05) for starter intake, with calves on 28/20 milk replacer consuming the greatest amounts after week 3 and maintaining the greater starter intake through week 8. A treatment over time effect was observed for fecal scores (P < 0.05). No other differences were observed (P > 0.05) in fecal scores among any treatments. Average wither heights were greater (P < 0.05) in calves fed 28/20 milk replacer. There were no effects (P > 0.05) of treatment on IGF-I concentrations, rumen pH, or concentrations of VFA. Acetate, as a percent of total VFA, tended (P < 0.10) to be greater in calves fed 28/20 milk replacer. These data indicate that increasing the dietary protein content of milk replacer without reducing fat content improved growth in young dairy calves without compromising health or rumen development.

Three oryx species and nine of eighteen antelope species across three tribes are considered endangered by the IUCN. Though the common eland (Taurotragus oryx) is not endangered, it lends itself well to the adaption of domestic assisted reproductive techniques due to its large size and calm temperament (Hansen et al., 1985). For these techniques to be used efficiently however, the details of the estrous cycle need to be defined within this and other species. Previously, Nowak (1999) proposed the eland estrous cycle to be ~21 days. In addition to cycle length, specifics of the estrous cycle have yet to be defined and are the objectives of this project. The overall objective is the characterization of the common eland estrous cycle. Specific objectives were to (1) observe behavior peri-estrus (2) determine ovulatory follicle size and (3) produce hormone profiles. The study animals consisted of two groups of four eland females (n=8) housed as a single bachelorette herd at ACRES. Each group was administered both of two commercial estrus synchronization regimens: Lutalyse® protocol (Regimen 1) and altrenogest protocol (Regimen 2). Regimen 1 received PGF2á injections on day 0 and day 11. Regimen 2 received altrenogest for 7 days and given PGF2á on day 7. After regimen administration intensive sample collections were performed around time of expected estrus and then repeated at expected time of subsequent natural estrus: blood samples were taken every 12 hours and ultrasonography was performed every 24 hours until the disappearance of a large follicle. HeatWatch® patches were applied to detect mounting behavior. Mounts were recorded least often during late morning (0600 to 1200). Dominant and ovulatory follicle size was determined to be 7 to 10 mm in diameter and estrous cycle duration was 21 ± 1.6 days. A total of seven of 30 possible ovulations (23.3%) was detected across both regimens. Homosexual mounting behavior was recorded, though all mounts were 2 seconds or less. Although further work needs to be conducted to confirm, the parameters defined here should help in the application of reproductive techniques to nondomestic ungulates.

Nuclear transfer (NT) results in low success rates and research has reported problems with the welfare of cloned animals produced. Evidence suggests the use of a less differentiated donor cell, such as a stem cell, may lead to an increase in the efficiency of the NT procedure. Stem cell characteristics must be clearly defined in order to isolate, identify and optimally culture potential donor cells. Stem cell characteristics such as self-renewal and differentiation have not been identified in stem cells from porcine adipose tissue, an easily attainable tissue resource. The phenotypic profile of cell surface proteins and chromosomal stability of porcine adipose tissue-derived stem cells (pASCs) through in vitro culture have not been described. Also, research has not identified epigenetic modifications in adult somatic mesenchymal stem cells (MSCs) from porcine adipose tissue. The objective of Project 1 was to develop a protocol for the isolation and culture of pASCs and to determine stem cell-like characteristics. The objective of Project 2 was to characterize the surface protein phenotype of undifferentiated porcine ASCs and to determine the chromosomal stability through in vitro culture. The objective of Project 3 was to analyze pASCs and compare them to fetal porcine fibroblasts (FPF) for gene expression profiles of chromatin remodeling proteins, global methylation and acetylation patterns and methylated lysine residues within promoter regions of developmentally important transcription factors. Improved culture conditions and defined cellular characteristics of these pASCs have been identified. pASCs can self-renew and differentiate into multiple tissue lineages. Porcine ASCs do not express the stem cell surface markers analyzed at similar levels compared with those of human ASCs reported in the literature. Also, in vitro-cultured porcine ASCs used as donor cells for NT should be chosen from early population doublings (PDs) due to increased levels of aneuploidy at later PDs. Further characterization of porcine ASCs in this manner could assist the isolation and purification of a population of MSCs from the easily obtainable tissue of adipose that can later be tested as donor cells in nuclear transfer.

Gastrointestinal nematode (GIN) parasitism is a problem for small ruminant producers. Haemonchus contortus is the most detrimental because of substantial economic losses. Over the past decades, chemical anthelmintics have been used to control GINs. Recently, GINs have developed resistance to most anthelmintics, and now reliance on anthelmintics is not possible. Alternative control methods are needed. Also, the use of fewer chemicals in agricultural products has called for alternative methods to be less synthetic and more organic. One promising alternative method is the feeding of condensed tannin (CT) containing forages as either fresh forage or dried products such as hay, ground hay, and pellets. Studies have shown that grazing or feeding sericea lespedeza (SL), a forage plant high in CTs, as whole plant resulted in some level of controlling H. contortus infection. The leaf has the highest CT content; so this study was conducted to determine which percentage of SL leaf meal was the most effective at controlling H. contortus infection in lambs and to determine any effect on reducing GIN larval development in the feces. Thirty-two cross bred lambs were randomly allocated into 4 groups with 8 animals each for a 5 week confinement trial. One group served as the control and received no SL. The other 3 groups received diets containing 25, 50, and 75% SL. Each week, feces and blood were collected to monitor infection level based on fecal egg count (FEC), blood packed cell volume (PCV), and enumeration and identification of larvae from fecal cultures. Fecal egg count decreased significantly (P < 0.05) over time for all 4 groups; however, there were no significant differences between groups. There was a trend on regression analysis for the control group FEC to be significantly (P < 0.001) greater during the trial. Reduction in FEC for the 3 treatment groups was 37.44-67.28% subsequent to week 1. There was no effect on PCV or nematode larvae population distribution and viability in feces. The antiparasitic effect of SL fed as leaf meal was not conclusive in this trial, but there was some evidence that suggested further evaluation is warranted.

Since the cloning of Dolly there has been little change in the efficiency of nuclear transfer (NT). Research is beginning to investigate the characteristics of donor cells. Adiposetissue is an abundant source of adult-derived cells that have displayed stemness in-vitro(Gimble et al., 2003). The overall goal of this research was to define the in-vitro characteristicsof bovine adipose-derived adult stem cells (ADAS) for the use in NT. Isolation methods weredetermined by a 3 x 3 factorial design. 1 g of subcutaneous fat was collected and subjected to0.10%, 0.25% or 0.50% collagenase type I solution for 1, 2 and 3 h. Nucleated cells werecounted using heochst stain. There was no significant difference (P>0.05) in number of nucleated cells released during the incubation period or collagenase concentrations. Viable cells were determined by those that remained adherent 24 h post plating. Incubation in 0.25% collagenase for 2 h had the consistently highest percentage of viable cells (45%). The lifespan and growth characteristics were determined by in a 2 x 2 factorial experiment of DMEM or DMEM:F12 supplemented or not supplemented with growth factors. DMEM with growth factors supplementation was significantly shorter lifespan (P>0.05) than DMEM:F12. The averagelifespan was ~30 population doublings (PDs), with 1 cell cycle every two days until passage 8(P8). Two bovine ADAS cell lines were differentiated into adipocytes, chondrocytes and osteoblasts at middle and late passages along side of adult derived skin fibroblasts. Differentiation was confirmed by histological staining resulting in early passage ADAS cells staining more intensely compared to late passage ADAS cells and skin fibroblasts. Global levels of DNA methylation and histone acetylation were analyzed from P1 to P6 in ADAS and skin fibroblasts from three animals. There was no significant difference (P>0.05) between cell types for DNA methylation or histone acetylation. The percentage of cleaved and developing blastocyst embryos from the ADAS cells (62% and 8%) and skin fibroblasts cells (42% and 8%)were not different (P>0.05). Interspecies nuclear transfer utilized eland ADAS cells into enucleated bovine oocytes. A total of 3 interspecies embryos (1%) developed to blastocyst.

The purpose of this research was to determine the state and regional mineral status of Louisiana forages and beef cattle. Louisiana beef cattle operations (n = 25) were sampled and divided into seven geographical regions, including the northwest (NW), northeast (NE), central (CE), southwest (SW), south central (SC), Florida parishes (FP) and southeast (SE) regions. Over a two year period, water and soil samples were collected from each operation annually, forage samples were collected quarterly in Aug to Sep, Nov to Dec, Feb to Mar and May to June and bovine serum samples were collected twice annually in the fall and spring seasons. The highest (P < 0.05) average regional water K and S concentrations were observed in the SE region and water Ca and Mg concentrations were the highest (P < 0.05) in the NE, CE and SE regions. However, all water mineral concentrations, with exception of Na, were lower than the reported upper desired levels considered safe for livestock consumption (Socha et al., 2003). Similar to water, soil Ca, Mg and K concentrations in our study, were higher (P < 0.05) in the SE compared to all other regions. Soil Cu concentrations were below critical levels in the CE region and all soil Zn concentrations, except the SE region, were lower than reported critical levels indicating soil deficiency. The average forage concentration for each mineral were: Ca (0.42%), P (0.28%), Mg (0.21%), K (1.83%), Na (0.10%), S (0.32%), Cu (8.12 ppm), Fe (323.46 ppm), Mn (254.85 ppm) and Zn (41.29 ppm). In addition, only mean forage Cu concentrations were lower than minimum requirements and regional forage K (NW region), Mg (FP region), Na (CE region) and S (NW and SE regions) concentrations were higher (P < 0.05) than other regions. The average regional serum K concentration in the NE region was higher (P < 0.05) than all other regions. Average bovine serum mineral concentrations in Louisiana were: Ca (9.02 mg/100 ml), P (13.62 mg/100 ml), Mg (1.92 mg/100 ml), K (21.66 mg/100 ml), Na (303.30 mg/100 ml), S (103.31 mg/100 ml), Cu (0.63 µg/ml), Fe (7.44 µg/ml), Zn (1.28 µg/ml), Mn (8.08 ng/ml) and Se (64.48 ng/ml). Furthermore, of these minerals, serum Mg, Na, Cu and Mn concentrations were lower than critical levels, indicative of deficiency.

A study was conducted to determine the effects of varying levels of cottonseed hulls on growth and metabolic indications of rumen development of dairy calves. Sixty-four Holstein calves (Heifers, n=40; Bulls, n=24) were randomly assigned to one of four dietary treatments which included calf starters containing no cottonseed hulls (control; C), 10% cottonseed hulls (10% CSH), 15% cottonseed hulls (15% CSH), or 20% cottonseed hulls (20% CSH). Calves were fed their respective treatments beginning on day 6 until day 56 of age. Body weights were measured at birth and biweekly thereafter until day 56 of age. Wither and hip heights were measured beginning on day 14 and biweekly thereafter until day 56 of age. Feed intake and fecal scores were recorded twice daily through day 56. On days 14, 28, 42, and 56, rumen fluid was collected for analysis of pH, volatile fatty acids (VFA), and ammonia (NH3), and blood was collected for analysis of plasma urea nitrogen (PUN) and â-hydroxybutyrate (BHBA). There was no treatment effect on average daily starter intake, body weight, wither height, and hip height but a treatment effect on fecal scores. Calves consuming CSH had higher rumen pH than C. Rumen pH also decreased as calves aged. There was no treatment effect on rumen acetate, propionate, butyrate, and total VFA concentrations. A treatment effect on NH3 concentrations was observed, and NH3 concentrations decreased over time. There was no treatment effect on BHBA, but a main effect of sex was observed in which the males had greater BHBA levels. There was no treatment effect on PUN concentrations, but a main effect of sex was observed with females having greater PUN concentrations. Overall, incorporating cottonseed hulls into a calf starter showed no significant effect on growth and rumen development in Holstein dairy calves.

This study investigated the effects of semen extender constituents and cryopreservation on bovine spermatozoal mitochondrial function. Three yearling Holstein bulls were used. Two ejaculates per bull were collected and pooled on a weekly basis for five weeks and extended in four treatments: 1) sodium citrate egg yolk extender with antibiotics (lincomycin, spectinomycin, gentamicin and tylosin); 2) ¡°1¡± with glycerol; 3) ¡°2¡± without antibiotics; and 4) ¡°1¡± without antibiotics. Each was divided into portions for analyses before freezing and after cryopreservation. The pre-freeze and thawed semen were transferred to a 37¡ãC water bath, the same assays were performed. In experiment 1, resazurin reduction (RD) was measured spectrophotometrically at sequential 25 minute intervals for 125 minutes. In experiment 2, specific activities of cytochrome c oxidase (CytoCox) and citrate synthase (CS) were measured spectrophotometrically immediately post-thaw and after 125 minutes of incubation. In experiment 3, ATP was measured using luciferin-luciferase assay simultaneously with RD. Total and progressive motilities (TM and PM), progressive (PV), curvilinear (VCL) and pathway (VAP) velocities were measured simultaneously with RD, ATP content and CytoCox and CS using computer assisted semen analysis system (CASA). In experiment 4, the NADH dehydrogenase (ND1) gene of mtDNA was sequenced before and after cryopreservation using PCR. Data were analyzed by least square methods; mean differences were delineated by Tukey¡¯s test. In experiment 1, RD differed among treatments (P<0.05). Cryopreservation decreased (P<0.05) RD, TM, PM, PV, VAP and VCL. Resazurin reduction correlated with PM (r=0.45, P<0.05) and TM (r=0.2, P<0.05). In experiment 2, incubation time and incubation with Triton X100 were sources of variation in CytoCox and CS specific activities (P>0.05). Only CS from spermatozoa incubated with Triton X100 correlated with RD (r=0.22, P<0.05). CytoCox and CS did not correlate with motility parameters. In experiment 3, spermatozoal ATP was not different (P>0.05) among treatments. However, cryopreservation decreased (P<0.05) ATP. Spermatozoal ATP correlated with motility parameters (r¡Ý0.65) and RD (r=-0.30) (P<0.05). In experiment 4, the frequency of amino acid change was higher (P<0.05) post-thaw in the treatment containing only antibiotics. Cryopreservation, more than extender constituents impacted mitochondrial function of bovine spermatozoa.

Gastrointestinal nematode parasites cause extensive damage to small ruminants, and Haemonchus contortus is a major concern to production. In the past, small ruminants were dewormed at regular intervals and control methods were based primarily on the use of anthelmintics. At present, anthelmintic resistance has been reported worldwide and has developed into a serious problem for small ruminant management programs. In view of this, alternate control methods are needed. One alternative method is the use of copper oxide wire particles (COWP). Three independent trials were conducted during Spring, Summer, and Fall of 2008. Two trials evaluated the effect of COWP in food pellets in reducing H. contortus infection in crossbred ewes and Suffolk lambs. The third trial compared the effect of Copasure® COWP with that of an industrial-grade COWP. Trials consisted of similar protocols where ewes and lambs were allocated into groups based on fecal egg count (FEC). Copasure® COWP in feed pellets were fed to ewes at mid-lactation when FEC increased to over 1000 eggs per gram, and to individual lambs when FAMACHA© score was 4/5. Ewe/lamb infection was monitored weekly by FEC and blood packed cell volume. Results of Trial 1 indicated that Copasure® COWP in feed pellets was effective in reducing the peri-parturient rise in ewes, based on FEC. Trial 2 indicated that Copasure® COWP in feed pellets was just as good as the levamisole/albendazole treatment when the FAMACHA System© was used to determine when to treat lambs. Trial 3 indicated that Copasure® COWP effectively reduced parasite infection while the industrial-grade COWP did not. The results from these trials demonstrated that the use of Copasure® COWP in feed pellets reduced H. contortus infection and may be useful alone or when used with other control methods.