Life Sciences Commons^™

Transmission Of Ranavirus Between Ectothermic Vertebrate Hosts, R Brenes, Matt J. Gray, T B. Waltzek, Rebecca P. Wilkes, Debra L. Miller

Debra L Miller

Transmission is an essential process that contributes to the survival of pathogens. Ranaviruses are known to infect different classes of lower vertebrates including amphibians, fishes and reptiles. Differences in the likelihood of infection among ectothermic vertebrate hosts could explain the successful yearlong persistence of ranaviruses in aquatic environments. The goal of this study was to determine if transmission of a Frog Virus 3 (FV3)-like ranavirus was possible among three species from different ectothermic vertebrate classes: Cope's gray treefrog (Hyla chrysoscelis) larvae, mosquito fish (Gambusia affinis), and red-eared slider (Trachemys scripta elegans). We housed individuals previously exposed to the FV3-like ranavirus …

Kisspeptin Receptor Agonist (Ftm080) Increased Plasma Concentrations Of Luteinizing Hormone In Anestrous Ewes, Brian K. Whitlock, Kelly A. Chameroy, Rebecca R. Payton, Shinya Oishi, Nobutaka Fujii, Joseph A. Daniel

Brian K Whitlock, PhD, DVM, DACT

Kisspeptin (KP) and the KP receptor (Kiss1r) are integral to central regulation of the gonadotropic-axis. The demonstration that intravenous infusion of KP can stimulate gonadotropin secretion and ovulation in seasonally anestrous female sheep offers a means of manipulating the reproductive axis. However, KP may be of limited clinical use because of the short circulating half-life. Kiss1r agonists with increased half-life and similar efficacy to KP in vitro potentially may provide beneficial applications in breeding management of many species. However, many of these agonists have not been tested in vivo. This study was designed to test and compare the effects of …

Effect Of Meloxicam On Gain And Inflammatory Response Of Calves Castrated By Banding Post-Weaning, Brian K. Whitlock, Peter D. Krawczel, Jeff A. Carroll, Nicole C. Burdick Sanchez, Jeff W. Dailey, Joseph A. Daniel, Johann F. Coetzee

Brian K Whitlock, PhD, DVM, DACT

Castration may detrimentally affect the health and performance of weaned calves, and painful procedures are increasingly becoming a public concern. The objective of this study was to determine the effects of castration (by banding) with or without administration of meloxicam, a non-steroid anti-inflammatory, on performance and inflammatory response in weaned beef calves. Forty-eight (weaned) beef calves [10.0±0.2 mo old; 304±6 kg BW] were blocked by age, BW, wither height, scrotal circumference, and source, then randomly assigned to 1 of 3 treatments (n=16 calves per treatment: 1) intact bulls (BULL), 2) castration by banding (BAN), or 3) castration by banding with …

Effect Of Meloxicam On Gain And Behavior Of Calves Castrated By Banding Pre-Weaning, Joseph A. Daniel, Peter D. Krawczel, Brian K. Whitlock

Brian K Whitlock, PhD, DVM, DACT

The objective was to determine if oral meloxicam (M; a non-steroid anti-inflammatory) administered at castration of pre-weaning calves affected ADG or behavior. Prior to castration (14 d), Angus bulls were assigned to bull (BULL; n = 7; age 106 ± 6 d; BW = 174.2 ±7.7 kg; scrotal circumference 17.7 ± 0.4 cm), castrated (BAN; n = 12; age = 105 ± 5 d; BW = 144.5 ± 7.6 kg; scrotal circumference = 16 ± 0.4 cm) or castrated with meloxicam (BAN+M; n = 13; age = 121 ± 6 d; BW = 145.8 ± 6 kg; scrotal circumference = …

Meloxicam Mediates Short-Term Behavioral Changes Of Castrated Calves, Peter D. Krawczel, Jeff A. Carroll, Nicole C. Burdick Sanchez, Jeff W. Dailey, Joseph A. Daniel, Johann F. Coetzee, Brian K. Whitlock

Brian K Whitlock, PhD, DVM, DACT

Castration may detrimentally affect the health and performance of weaned calves and painful procedures are increasingly a public concern. Therefore, practical pain mitigation is critical. The objective was to determine the effects of castration (by banding) with or without administration of NSAID, meloxicam, on the behavior of weaned beef calves. Forty-eight (56 d post-weaning) beef calves [10.0 ± 0.2 (mean ± SE) mo old; 304 ± 6 kg BW] were blocked by multiple factors then randomly assigned to 3 treatments (n = 16 calves per treatment: 1) intact bulls (BULL), 2) castration by banding (BAN), or 3) castration by banding …

Avma Guidelines For The Euthanasia Of Animals: 2013 Edition, Steven Leary, Wendy Underwood, Raymond Anthony, Samuel Cartner, Douglas Corey, Temple Grandin, Cheryl B. Greenacre, Sharon Gwaltney-Bran, Mary Ann Mccrackin, Robert Meyer, David Miller, Jan Shearer, Roy Yanong

Cheryl B Greenacre DVM

No abstract provided.

Heifer Estrous Cycle Stage Impacts Luteinizing And Growth Hormone Response To Kisspeptin, Brian K. Whitlock, Joseph A. Daniel, Robyn R. Wilborn, Barbara P. Steele, James L. Sartin

Brian K Whitlock, PhD, DVM, DACT

The reproductive neuropeptide, kisspeptin (KP), may have a role in regulation of growth hormone (GH) secretion. Expression of KP and its receptor (Kiss1r) are developmentally and hormonally regulated. This study was conducted to determine the effects of KP on plasma concentrations of luteinizing hormone (LH) and GH in pubertal heifers during different functional reproductive stages. Heifers received a single injection of KP (100 pmole / kg) or saline intravenously during proestrus, estrus, and diestrus, and serial blood samples were collected for 3 hours to determine the response of both LH and GH after treatment. Mean concentrations of LH were increased …

Effect Of Kisspeptin On Regulation Of Growth Hormone, Brian K. Whitlock, Joseph A. Daniel, Robyn R. Wilborn, Herris S. Maxwell, Barbara P. Steele, James L. Sartin

Brian K Whitlock, PhD, DVM, DACT

Kisspeptin ( KP ), a neuroendocrine regulator of reproduction, is hypothesized to be an integrator of metabolism and hormones critical to the regulation of reproduction. Lactation is associated with enhanced growth hormone (GH) responsiveness and reduced fertility. Our study was designed to determine the effects of lactation on KP-stimulated GH and luteinizing hormone (LH) secretion. Five non-lactating and five lactating dairy cows were used in the study. Experiments were conducted with lactating cows at weeks 1, 5 and 11 after parturition. The experimental treatments ( saline and KP [100 and 400 pmol / kg body weight]) were given intravenously and …

The Effect Of Kisspeptin Receptor Agonist (Ftm080) On Luteinizing Hormone In Sheep, Brian K. Whitlock, Joseph A. Daniel, Maria Hes, Barbara P. Steele, James L. Sartin, S. Oishi, N. Fujii

Brian K Whitlock, PhD, DVM, DACT

Kisspeptin receptor (Kiss1r) agonists with increased half-life and similar efficacy to kisspeptin (KP) in vitro potentially may provide beneficial applications in breeding management of many species. However, many of these agonists have not been tested in vivo. This study was designed to test the effect of a Kiss1r agonist (FTM080) on luteinizing hormone (LH) in vivo. Sheep were treated with FTM080 (500 pmol/kg BW) or sterile water in a 2-ml bolus via the jugular cannula. Serial blood samples were collected every 15-min before (1 hr) and after (1 hr) treatment. Intravenous (IV) injection of FTM080 increased (P < 0.05) LH plasma concentrations through the 45-min sample following treatment. Moreover, the area under the curve of LH in the period from 0 to 60-min following FTM080 treatment was also increased (P < 0.05). These data provide evidence to suggest that FTM080 stimulates the gonadotropic axis of ruminants in vivo.

Method For Somatic Cell Nuclear Transfer In Zebrafish, Kannika Siripattarapravat, Jose Cibelli

Jose Cibelli

Somatic cell nuclear transfer (SCNT) has been a well-known technique for decades and widely applied to generate identical animals, including ones with genetic alterations. The system has been demonstrated successfully in zebrafish. The elaborated requirements of SCNT, however, limit reproducibility of the established model to a few groups in zebrafish research community. In this chapter, we meticulously outline each step of the published protocol as well as preparations of equipments and reagents used in zebrafish SCNT.

Selected Hormonal And Neurotransmitter Mechanisms Regulating, James L. Sartin, Joseph A. Daniel, Brian K. Whitlock, Robyn R. Wilborn

Brian K Whitlock, PhD, DVM, DACT

Appetite control is a major issue in normal growth and in suboptimal growth performance settings. A number of hormones, in particular leptin, activate or inhibit orexigenic or anorexigenic neurotransmitters within the arcuate nucleus of the hypothalamus, where feed intake regulation is integrated. Examples of appetite regulatory neurotransmitters are the stimulatory neurotransmitters neuropeptide Y (NPY), agouti-related protein (AgRP), orexin and melanin-concentrating hormone and the inhibitory neurotransmitter, melanocyte-stimulating hormone (MSH). Examination of messenger RNA (using in situ hybridization and realtime PCR) and proteins (using immunohistochemistry) for these neurotransmitters in ruminants has indicated that physiological regulation occurs in response to fasting for several …

Effect Of Leukemia Inhibitory Factor On Feed Intake And Body Temperature In Sheep, James L. Sartin, D L. Marks, Brian K. Whitlock, Joseph A. Daniel, Barbara P. Steele

Brian K Whitlock, PhD, DVM, DACT

Leukemia inhibitory factor (LIF) has been suggested to function as a potent inhibitor of feed intake in rodents. These studies were designed to determine whether LIF was found in the ovine hypothalamus and whether LIF inhibited feed intake in sheep. Sheep hypothalami were used to clone LIF to indicate presence of the gene in the hypothalamus. The sequence was similar to published data. Another group of sheep were provided intraventricular (ICV) cannulas and injected with doses of LIF at 250, 500, 1000 and 2500 ng per sheep, ICV. Feed intake was inhibited by the 1000 and 2500 ng dose (trt, …

Changes In Plasma Concentrations Of Growth Hormone And Luteinizing Hormone In Ewes Following Central And Peripheral Treatment With Kisspeptin, Brian K. Whitlock, Joseph A. Daniel, Barbara P. Steele, James L. Sartin

Brian K Whitlock, PhD, DVM, DACT

Kisspeptin (KP), a neuroendocrine regulator of gonadotropin releasing hormone, has been hypothesized as an integrator of nutrition and hormones critical to metabolism and regulation of reproduction. Recent evidence suggests growth hormone (GH) secretion may be influenced by KP. The objective of this study was to determine if the GH stimulatory effect of KP is due to actions on the hypothalamus or anterior pituitary gland in ewes. Adult ovariectomized ewes (n=8) were fitted with intracerebroventricular (ICV) cannula to facilitate central administration of experimental treatments. Ewes received one of eight treatments [four intravenously (IV) and four ICV]. Peripheral treatments [0 (Veh), 100, …

Changes In Plasma Concentrations Of Leptin In Ewes During Pregnancy, Joseph A. Daniel, A B. Milam, M E. Gafnea, Brian K. Whitlock, D H. Keisler

Brian K Whitlock, PhD, DVM, DACT

Previous research has demonstrated circulating concentrations of leptin increase in ewes during mid pregnancy then decline in late pregnancy and early lactation. The objective of this study was to more narrowly define the timing of changes in circulating concentrations of leptin with pregnancy in ewes. Katahdin ewes (n= 19) located at latitude 34.275 and longitude -85.183 (Mount Berry, GA) were utilized. Blood samples were collected weekly via jugular veinpuncture beginning immediately before ram exposure on September 23 and continuing until 4 weeks post-lambing. Ewes were exposed to a ram fitted with a marking harness for a 63 day breeding season. …

Farm Focus - Spring 2010, Brian K. Whitlock, Matt Welborn, Maria Prado, Amy Plummer

Brian K Whitlock, PhD, DVM, DACT

No abstract provided.

Ppar Agonists Down-Regulate The Expression Of Atp10c Mrna During Adipogenesis, A Peretich, Maria Cekanova Ms, Rndr, Phd, S Hurst, Sj Baek, Madhu Dahr

Maria Cekanova MS, RNDr, PhD

No abstract provided.

Somatic Cell Nuclear Transfer In Zebrafish, Kannika Siripattarapravat, Patrick J. Venta, C C. Chang, Jose B. Cibelli

Jose Cibelli

We developed a method for somatic cell nuclear transfer in zebrafish using laser-ablated metaphase II eggs as recipients, the micropyle for transfer of the nucleus and an egg activation protocol after nuclear reconstruction. We produced clones from cells of both embryonic and adult origins, although the latter did not give rise to live adult clones.

Farm Focus - Spring 2009, Brian K. Whitlock, Matt Welborn, Maria Prado, Amy Plummer

Brian K Whitlock, PhD, DVM, DACT

No abstract provided.

Farm Focus - Fall 2009, Brian K. Whitlock, Matt Welborn, Maria Prado, Amy Plummer

Brian K Whitlock, PhD, DVM, DACT

No abstract provided.

The Effects Of Lactation And Negative Energy Balance On Kisspeptin-Stimulated Luteinizing Hormone And Growth Hormone In Dairy Cows, Brian K. Whitlock, Joseph A. Daniel, Robyn R. Wilborn, Herris S. Maxwell, Barbara P. Steele, James L. Sartin

Brian K Whitlock, PhD, DVM, DACT

Kisspeptin, a neuroendocrine regulator of gonadotropin releasing hormone, is hypothesized to integrate nutrition and hormones critical to metabolism and the regulation of reproduction. Since the negative energy balance of early lactation is associated with reduced fertility via suppression of gonadotropin secretion and enhanced growth hormone (GH) responsiveness, this experiment was designed to determine the effects of stage of lactation and negative energy balance on kisspeptin-10 (Kp-10) stimulated luteinizing hormone (LH) and GH concentrations. Five nonlactating [5.1 ± 0.8 (SEM) years; 577 ± 19 kg body weight (BW)] and five lactating [4.1 ± 0.6 years; 608 ± 11 kg BW] multiparous …

Expression Of G-Protein Inwardly Rectifying Potassium Channels (Girks) In Lung Cancer Cell Lines, Howard Plummer 3rd, Madhu Dhar, Maria Cekanova Ms, Rndr, Phd, Hildegard Schuller

Maria Cekanova MS, RNDr, PhD

BACKGROUND: Previous data from our laboratory has indicated that there is a functional link between the beta-adrenergic receptor signaling pathway and the G-protein inwardly rectifying potassium channel (GIRK1) in human breast cancer cell lines. We wanted to determine if GIRK channels were expressed in lung cancers and if a similar link exists in lung cancer. METHODS: GIRK1-4 expression and levels were determined by reverse transcription polymerase chain reaction (RT-PCR) and real-time PCR. GIRK protein levels were determined by western blots and cell proliferation was determined by a 5-bromo-2'-deoxyuridine (BrdU) assay. RESULTS: GIRK1 mRNA was expressed in three of six small …

Multiple Luteinizing Hormone Receptor (Lhr) Protein Variants, Interspecies Reactivity Of Anti-Lhr Mab Clone 3b5, Subcellular Localization Of Lhr In Human Placenta, Pelvic Floor And Brain, And Possible Role For Lhr In The Development Of Abnormal Pregnancy, Pelvic Floor Disorders And Alzheimer's Disease, A Bukovsky, K Indrapichate, H Fujiwara, Maria Cekanova Ms, Rndr, Phd, Me Ayala, R Dominguez, Mr Caudle, J Wimalsena, Rf Elder, P Copas, Jf Foster, Ri Fernando, Dc Henley, Nb Upadhyaya

Maria Cekanova MS, RNDr, PhD

Distinct luteinizing hormone receptor (LHR) protein variants exist due to the posttranslational modifications. Besides ovaries, LHR immunoreactivity (LHRI) was also found in other tissues, such as the brain, fallopian tube, endometrium, trophoblast and resident tissue macrophages. The 3B5 mouse monoclonal antibody was raised against purified rat LHR. In rat, porcine and human ovaries, the 3B5 identified six distinct LHR bands migrating at approximately 92, 80, 68, 59, 52 and 48 kDa. Characteristic LHRI was detected in rat, human and porcine corpora lutea. During cellular differentiation, subcellular LHR distribution changed from none to granular cytoplasmic, perinuclear, surface, nuclear and no staining. …

Expression And Localization Of Estrogen Receptor-Alpha Protein In Normal And Abnormal Term Placentae And Stimulation Of Trophoblast Differentiation By Estradiol, A Bukovsky, Maria Cekanova Ms, Rndr, Phd, Mr Caudle, J Wimalasena, Js Foster, Dc Henley, Rf Elder

Maria Cekanova MS, RNDr, PhD

Estrogens play an important role in the regulation of placental function, and 17-beta-estradiol (E2) production rises eighty fold during human pregnancy. Although term placenta has been found to specifically bind estrogens, cellular localization of estrogen receptor alpha (ER-alpha) in trophoblast remains unclear. We used western blot analysis and immunohistochemistry with h-151 and ID5 monoclonal antibodies to determine the expression and cellular localization of ER-alpha protein in human placentae and cultured trophoblast cells. Western blot analysis revealed a ~65 kDa ER-alpha band in MCF-7 breast carcinoma cells (positive control). A similar band was detected in five normal term placentae exhibiting strong …

Characterization Of A Piscirickettsiosis-Like Disease In Hawaiian Tilapia., M. J. Mauel, Debra L. Miller, K. Frazier, A. Liggett, E. Styer, D. Montgomery-Brock, J. Brock

Debra L Miller

In 1994, tilapia (Oreochromis mossambicus and Sarotherodon melanotheron) in wild and farmed populations on Oahu, Hawaii, USA, began to die of an unknown disease that was similar but not identical to piscirickettsiosis in salmonids. Only tilapia were affected. Diseased tilapia often swam erratically and had trouble staying at depth. Scattered cutaneous haemorrhage and exophthalmia were often noted. In many cases, fish were found dead with no clinical signs. Gills exhibited epithelial hyperplasia with severe multifocal consolidation of secondary lamellae. Multiple granulomas were observed in the gills, spleen, kidney, choroid gland and testes, but not in the liver. Tilapia mortalities occurred …

Placental Expression Of Estrogen Receptor Beta And Its Hormone Binding Variant – Comparison With Estrogen Receptor Alpha And A Role For Estrogen Receptors In Asymmetric Division And Differentiation Of Estrogen-Dependent Cells, Antonin Bukovsky, Michael R. Caudle, Maria Cekanova Ms, Rndr, Phd, Romaine I. Fernando, Jay Wimalasena, James S. Foster, Donald C. Henley, Robert F. Elder

Maria Cekanova MS, RNDr, PhD

During human pregnancy, the production of 17-beta-estradiol (E2) rises steadily to eighty fold at term, and placenta has been found to specifically bind estrogens. We have recently demonstrated the expression of estrogen receptor alpha (ER-alpha) protein in human placenta and its localization in villous cytotrophoblast (CT), vascular pericytes, and amniotic fibroblasts. In vitro, E2 stimulated development of large syncytiotrophoblast (ST) aggregates. In the present study we utilized ER-beta affinity purified polyclonal (N19:sc6820) and ER-alpha monoclonal (clone h-151) antibodies. Western blot analysis revealed a single ~52 kDa ER-beta band in chorionic villi (CV) protein extracts. In CV, strong cytoplasmic ER-beta immunoreactivity …

Short Communication: Relationship Between Body Growth And Mammary Development In Dairy Heifers, L F. Silva, M J. Vandehaar, Brian K. Whitlock, R P. Radcliff, H A. Tucker

Brian K Whitlock, PhD, DVM, DACT

Our objective was to determine if prepubertal rate of body weight (BW) gain, independent of diet, was related to mammary development of dairy heifers. Data from two studies recently conducted at Michigan State University were used to identify factors, within a dietary treatment group, that would account for variation in first lactation milk production or amount of mammary parenchymal DNA at the time of puberty. Factors analyzed for variation in milk production during first lactation were: postpartum BW, prepubertal BW gain, gestational BW gain, postpartum BW gain, body condition score (BCS) at breeding, and BCS at calving. Factors analyzed for …

Effect Of Dietary Protein On Prepubertal Mammary Development In Rapidly Growing Dairy Heifers, Brian K. Whitlock, M J. Vandehaar, L F. Silva, H A. Tucker

Brian K Whitlock, PhD, DVM, DACT

The objective was to determine whether increased dietary protein would enhance mammary development in prepubertal heifers fed for rapid body growth (1.2 kg/d). Fifty-four Holstein heifers (weighing approximately 134 kg) were assigned to one of three treatments. Heifers were fed a total mixed ration with metabolizable energy at 2.85 Mcal/kg and metabolizable protein at low, standard, or high concentrations (37, 41, or 44 g/Mcal of metabolizable energy, respectively) from 3.5 mo of age until slaughter at approximately 46 d after puberty. Heifers fed low, standard, and high protein gained 1130, 1170, and 1180 g/d, respectively. Dietary protein did not affect …

Variability Of Placental Expression Of Cyclin E Low Molecular Weight Variants, A Bukovsky, Maria Cekanova Ms, Rndr, Phd, Mr Caudle, J Wimalasena, Js Foster, Ja Keenan, Rf Elder

Maria Cekanova MS, RNDr, PhD

No abstract provided.

Abnormal Expression Of P27kip1 Protein In Levator Ani Muscle Of Aging Women With Pelvic Floor Disorders – A Relationship To The Cellular Differentiation And Degeneration, Antonin Bukovsky, Pleas Copas, Michael R. Caudle, Maria Cekanova Ms, Rndr, Phd, Tamara Dassanayake, Bridgett Asbury, Stuart E. Van Meter, Robert F. Elder, Jeffrey B. Brown, Stephanie B. Cross

Maria Cekanova MS, RNDr, PhD

Background Pelvic floor disorders affect almost 50% of aging women. An important role in the pelvic floor support belongs to the levator ani muscle. The p27/kip1 (p27) protein, multifunctional cyclin-dependent kinase inhibitor, shows changing expression in differentiating skeletal muscle cells during development, and relatively high levels of p27 RNA were detected in the normal human skeletal muscles.

Methods Biopsy samples of levator ani muscle were obtained from 22 symptomatic patients with stress urinary incontinence, pelvic organ prolapse, and overlaps (age range 38–74), and nine asymptomatic women (age 31–49). Cryostat sections were investigated for p27 protein expression and type I (slow …