Browsing by Author "Sponenberg, D. Phillip"

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    A Case of Ovarian Dysplasia and a Vaginal Fibroleiomyoma in a Young Golden Retriever
    McCarter, Samantha; Sponenberg, D. Phillip; Saunders, Geoffrey; Cecere, Julie (Frontiers, 2021-12-24)
    This case demonstrates a unique ovarian congenital anomaly that likely contributed to the development of a rare fibroleiomyoma in the cranial vagina of a young bitch. A 13 month old intact female Golden Retriever presented to the veterinary teaching hospital for urinary incontinence, hematuria, and persistent vaginal discharge. Physical examination revealed a mucopurulent serosanguinous malodorous vulvar discharge, and after further diagnostics was reclassified as persistent estrus. Abdominal palpation and ultrasound revealed uterine thickening and poorly visualized ovaries. The reproductive tract was removed during an ovariohysterectomy, revealing small ovaries and a white anterior vaginal mass. Histopathology revealed dysplastic ovaries with hyperplastic granulosa cells and a benign vaginal fibroleiomyoma. These morphologic changes are consistent with elevated estrogen levels. It was thus concluded that her persistent estrus and the fibroleiomyoma were both secondary to persistent estrogen production by the hyperplastic granulosa cells.
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    A Case of Segmental Aplasia of the Uterus, Cervix, and Cranial Vagina in a Cat
    Souther, Samantha; Baik, Nam Joo; Clapp, Kemba; Nappier, Michael T.; Sponenberg, D. Phillip; Cecere, Julie T. (Frontiers, 2019-05-15)
    This case documents a rare set of congenital anomalies that resulted in an atypical cystic lesion in the cranial vagina of a queen. A discrete cystic lesion was identified in an 8 year old intact female domestic shorthair cat presenting for routine ovariohysterectomy. Morphological, radiographic, and histopathological findings were consistent with segmental aplasia of the uterus, cervix, and vagina resulting in a blind dilation of the cranial vagina. Segmental vaginal aplasia in combination with the failed canalization of the cervix resulted in a blind portion of the cranial vagina, in which normal secretions collected and became inspissated. This formed a discrete cystic structure. This case represents a novel combination and clinical presentation of segmental aplasia in the cat, involving both the uterus and cranial vagina. Ovariohysterectomy was performed for sterilization and a partial vaginectomy was performed to remove the cystic lesion in its entirety.
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    Characterization of the Expression of Angiogenic Factors in Cutaneous Squamous Cell Carcinoma of Domestic Cats
    Gudenschwager-Basso, Erwin Kristobal; Stevenson, Valentina; Sponenberg, D. Phillip; Cecere, Thomas E.; Huckle, William R. (MDPI, 2022-07-21)
    Cutaneous squamous cell carcinoma (CSCC) is a common malignant skin cancer with a significant impact on health, and it is important to determine the degree of reliance of CSCC on angiogenesis for growth and metastasis. Major regulators of angiogenesis are the vascular endothelial growth factor (VEGF) family and their associated receptors. Alternative pre-mRNA splicing produces multiple isoforms of VEGF-A and PLGF with distinct biological properties. Several studies highlight the function of VEGF-A in CSCC, but there are no studies of the different isoforms of VEGF-A and PLGF for this neoplasm. We characterized the expression of three isoforms of VEGF-A, two isoforms of PLGF, and their receptors in cat CSCC biopsies compared to normal haired skin (NHS). Although our results revealed no significant changes in transcript levels of panVEGF-A or their isoforms, the mRNA levels of PLGF I and the receptors Flt-1 and KDR were downregulated in CSCC compared to NHS. Differences were observed in ligand:receptor mRNA expression ratio, with the expression of VEGF-A relative to its receptor KDR higher in CSCC, which is consistent with our hypothesis and prior human SCC studies. Immunolocalization in tissue showed increased expression of all measured factors and receptors in tumor cells compared to NHS and surrounding vasculature. We conclude that the factors measured may play a pivotal role in CSCC growth, although further studies are needed to clarify the role of angiogenic factors in feline CSCC.
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    Conservation Strategies for Local Breed Biodiversity
    Sponenberg, D. Phillip; Martin, Alison; Couch, Charlene; Beranger, Jeannette (MDPI, 2019-09-27)
    Breeds of domesticated animals are often overlooked as contributing to biodiversity. Their unique role at the junction of natural and human-influenced environments makes them important potential contributors to biodiversity and ecosystem function. Effective conservation of livestock breeds rests upon valid recognition of breeds as repeatable genomic packages with a reasonably high level of predictability for performance. Local or landrace breeds that lack breeder organization are especially difficult to conserve due to lack of formal recognition as breeds. Achieving success with them involves three major steps: Discover, Secure, and Sustain. Early in the process an evaluation of candidate populations for status as genetic resources is essential. This process is aided by a phenotypic matrix which can be used alongside historical investigations and genetic (DNA) studies. The goal is to include all qualifying animals and to exclude all those that do not qualify. Securing some populations depends on careful rescue protocols for maximizing the recovery of genetic variation, and this can then be followed by breeding protocols that provide for maintaining the population’s production potential along with a healthy and viable genetic structure for long-term survival and use. Sustaining breeds for the long term is also enabled by assuring market demand for the breed and its products.
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    Conserving the Genetic Diversity of Domesticated Livestock
    Sponenberg, D. Phillip (MDPI, 2020-07-17)
    Domesticated animals live and produce in an environment influenced by both natural and human factors. These agricultural environments are important to maintain for human survival and also for their interactions with natural environments. Effective conservation of domesticated biodiversity can help to assure sustainable agricultural systems that minimize negative influences on natural environments. In addition, livestock biodiversity is a component of total biodiversity and for several species is the only remaining source of diversity because the wild ancestors are now extinct. Conservation of livestock biodiversity depends on cultural and biological approaches. Each of these has differential importance depending on the specific location of the genetic resource as well as the human culture in which it resides. Effective global conservation blends these in different measures to assure positive outcomes that succeed in securing the genetic resource as well as its contribution to human survival and well-being.
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    DNA Sequence and Haplotype Variation Analysis of Inflammatory Response Genes NLRX1, IL6, and IL8 in the Turkey (Meleagris gallopavo)
    Russell, Kadijah Lashunte (Virginia Tech, 2019-02-08)
    Genotype-phenotype analyses continue to be the primary goal for genome analyses in livestock and poultry breeding. Essential to accomplish this goal is the need to identify variation at the genomic level. To test the hypothesis that DNA sequence variations in inflammatory response genes are associated with phenotypic differences in the heritage turkey, the primary objective of this project was to search for single nucleotide polymorphisms (SNPs) in candidate inflammatory response genes. A minor objective was to develop a system for inducing inflammatory response in the turkey using a microbe-based lipopolysaccharide (LPS), an approach previously described for the chicken. A total of 18 SNPs was identified in the three genes screened in this project: Interleukin 6 (IL6) and 8 (IL8), and NLRX1. Mortality data from the LPS challenge were not significantly different among the strains. Further gene expression analyses will be part of future work. The SNP data represent the first extensive analyses of candidate inflammatory response genes in the turkey. Combined with the protocols developed for inflammation assessment in the turkey the SNPs described here will be valuable resources for future inflammation:genotype evaluation in the turkey
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    Efficacy of the Bromsulfophthalein (BSP) 30-Minute Retention Test for the Diagnosis of Hepatobiliary Disease in Dogs
    Flatland, Bente (Virginia Tech, 1997-06-13)
    Measuring the amount of bromsulfophthalein (BSP) retained in serum 30 minutes after intravenous injection can be used to evaluate hepatic function. BSP retention of less than 5% 30 minutes after intravenous administration is considered normal in dogs. The BSP retention test fell out of favor due to perceived inaccuracy when compared with bile acid and ammonia testing and a fear of hypersensitivity reactions. BSP was discontinued as a commercially-available drug in 1984. Use of BSP has continued at virginia tech despite the test's reputed disadvantages. The purpose of this study was to evaluate the efficacy of the BSP retention test in dogs with and without histopathologically-confirmed hepatobiliary disease. The medical records of 150 dogs with hepatobiliary disease having both a BSP retention test and hepatic biopsy performed were evaluated. Histopathologic slides were reviewed, and dogs were classified according to one of 11 predetermined histopathologic categories. Twenty-five random-source dogs were used as controls. Adverse effects following BSP administration were not observed in any dog. BSP retention was significantly different between hospitalized and control dogs, but the test could not distinguish between dogs with different types of hepatobiliary disease. Sensitivity, specificity, and predictive values of the BSP retention test were calculated, and its sensitivity was comparable to that of serum bile acid and ammonia testing. Using 6.0% retention as a cut-off for normal retention resulted in a specificity of 100%, sensitivity of 69.8%, positive predictive value of 100%, and negative predictive value of 35.7%.
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    Evaluating the Expression of Angiogenic Mediators in a Mouse Model of Tumor Metastasis
    Crawford, Natalie M. (Virginia Tech, 2008-07-03)
    Solid tumors typically require angiogenesis, the development of new blood vessels, for growth and metastasis. Vascular endothelial growth factor (VEGF) induces angiogenesis by activating receptors on host endothelial cells. One such receptor, Flt-1, occurs as either a membrane bound or a secreted form (sFlt-1) that can inhibit angiogenic signaling. Previous studies have shown that variation in mRNA expression of VEGF and its receptors KDR, sFlt-1 and Flt-1 occurs in pathological angiogenesis, i.e. metastatic tumorigenesis. We hypothesize that the ratio of sFlt-1:Flt-1 mRNA will be altered in the presence of solid tumors. The objective of this study was to evaluate the expression of sFlt-1 and Flt-1 mRNAs in a mouse metastatic tumor model using CT26.CL25 cells. CT26.CL25 cells are VEGF-producing murine colon carcinoma cells transfected with the lacZ gene, which expresses B-galactosidase activity. These cells, injected intravenously, form tumor nodules in the lung. A pilot study revealed development of lung nodules in mice nine days after intravenous injection with 105 cells. In a second study, twenty-five 10- week-old female Balb/c mice were injected intravenously, via tail vein, with 2 x 105cells, and fifteen with vehicle control. Lung nodules developed in all mice injected with cells. Tissues were harvested by routine necropsy and either formalin-fixed for routine histology/histochemistry or stored for quantitative RT-PCR (QPCR) analysis of gene expression. Under microscopic evaluation, sections of lungs stained with Hematoxylin & Eosin (H&E) revealed nodules composed of polygonal neoplastic cells. cDNA from lungs (14 tumor-bearing, 10 controls) and cultured CT26.CL25 cells was analyzed by QPCR using primers and TaqMan probes directed against sFlt-1, Flt-1, KDR, VEGFA, PlGF (Placental Growth Factor), Angiotensin Converting Enzyme (ACE), 18S ribosomal RNA and neoR (neomycin phosphotransferase). We observed an increased sFlt-1:Flt-1 ratio in tumor-bearing versus control lungs, suggesting that tumor-derived signals may influence sFlt-1 and Flt-1 expression differentially. Additionally, there was increased expression of Flt-1, sFlt-1 and KDR in tumors versus controls, but not in VEGF expression in tumors versus controls. Interestingly, expression of PlGF was increased in tumors versus controls, suggesting its role as an enhancer of tumor progression in the presence of other angiogenic factors. Together, these findings indicate that solid tumor angiogenesis results from an intricate balance of various angiogenic factors.
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    Evaluation of a Paratuberculosis Quantitative Polymerase Chain Reaction Assay with Microscopic Correlation
    Tyler, Ronald Dale Jr. (Virginia Tech, 2012-05-01)
    Paratuberculosis is an intestinal condition in ruminants infected with Mycobacterium avium subspecies paratuberculosis (MAP) and precedes Johne's disease, a chronic enteric disorder in ruminants caused by MAP infection. Necropsy with histopathology provides definitive diagnosis of Johne's disease and positive culture of MAP from tissues provides definitive diagnosis of paratuberculosis. To determine assay sensitivity, 85 formalin-fixed paraffin-embedded (FFPE) tissues from ruminants diagnosed with Johne's disease were tested with a commercial paratuberculosis quantitative polymerase chain reaction (qPCR) assay and had a sensitivity of 92%. To determine assay specificity, 21 FFPE tissues from animals without gastrointestinal disease combined with 13 FFPE tissues from non-ruminant animals (frog, dove, turtle, dog, and 2 cats) with non-paratuberculosis mycobacterial diseases were tested with the commercial qPCR assay and had a specificity of 100%. Slides prepared from the FFPE tissue blocks were stained with hematoxylin & eosin (H&E) and Ziehl-Neelsen's (acid fast stain), then examined for granulomatous inflammation and scored on a scale from 0-4 based on the quantity of acid fast bacteria (AFB). Digital microscopy and morphometric software were used to compute an acid fast bacteria area index (AFBAI) to evaluate a more precise correlation with the qPCR results. The quantity of AFB in tissue slides showed medium to strong correlation with the appropriate qPCR results. The results indicate that the commercial qPCR assay can be used on FFPE tissues with good results and the qPCR results have medium-strong correlation with quantitative acid fast histopathology.
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    Evaluation of the Normal Equine Pituitary Gland
    McKlveen, Tori Leigh (Virginia Tech, 2003-07-21)
    Computed tomography (CT) is becoming more available as a diagnostic tool in the evaluation of the equine skull and brain. Objectives of this study were: 1) refine a CT protocol for evaluating the equine pituitary gland, 2.) define the CT anatomy of the pituitary region, 3.) determine a set of normal values for the pituitary dimensions (length, width, height, volume and weight), 4.) refine CT techniques for measuring pituitary size. Horses were scanned using 10x10mm, 10x5mm, 4x4mm and 4x2mm slice thickness and interval combinations. The pituitary glands were removed immediately after CT and gross measurements were performed. CT measurements were compared with gross pituitary measurements using analysis of variance (ANOVA) in a randomized block design. Accuracy percentages were also calculated using gross measurements as the known value. Mean dimensions of the histologically normal pituitary glands were: length 21.07mm, width 21.62mm, height 9.78mm and volume 2.66cm³. The weights ranged from 1.7g to 3.4g with a mean of 2.6g. Computed tomographic measurement analysis demonstrated that the 10mm slices were the most accurate way to estimate the length of the gland. The 4mm slices yielded the highest accuracy values for width, height and volume of the pituitary gland. The volume was underestimated by all interval and slice thickness combinations performed by CT. No evidence of an overlap effect was identified for any of the dimensions. Our findings indicated that contrast-enhanced CT is an accurate technique for estimating pituitary linear dimensions. Three-dimensional CT volumetry may not be an accurate method for estimating pituitary volume.
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    Gene Mapping of Morphological Traits in Chickens
    Li, Jingyi (Virginia Tech, 2017-04-25)
    Chickens exhibit considerable variation in morphological traits, with some populations having undergone intensive selection for uniqueness and uniformity. These populations are a source of experimental material to study the genetics of morphological traits. An important first step in such studies is to map the genes and the causal mutations that influence these traits. This research focused on gene mapping of 12 morphological traits including 4 intra-feather color patterns (Pattern, Columbian, Melanotic, and mottling), 2 inter-feather color patterns (Blue and chocolate), 2 forms of feathered-legs, polydactyly, dark brown eggshell color, vulture hock, and creeper. Ten backcross and/or F2 populations were designed to produce 1,880 individuals. An additional 339 DNA samples from other populations were included. The procedures for gene mapping were: 1. Pooling of DNA samples of backcross or F2 individuals based on their shared phenotypes, followed by microarray assays for genotyping, a cost-effective initial screen for the candidate genomic regions, 2. Linkage mapping to narrow the range of candidate genes, 3. Sequencing to identify the candidate mutations, 4. Diagnostic tests to confirm the association between the candidate mutation and the phenotype. Of the 12 traits studied, 3 (mottling, Blue, and chocolate) made progress into step 4. Complexities due to genomic context, modifiers, and environmental factors precluded step 4 for the first form of the feathered-leg gene, step 3 for the mapping of Melanotic, and earlier stages for the mapping of Pattern, Columbian, dark brown egg, vulture hock, and the second form of feathered-leg. These findings provide insights of the complexity of how background genome can influence the phenotypic expression of single genes (gene genetic background interactions) and an understanding of cellular and molecular mechanisms involved in morphogenesis.
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    Genetic Analysis of Toxin-Induced Dilated Cardiomyopathy in the Turkey (Meleagris gallopavo)
    Gyenai, Kwaku Barima (Virginia Tech, 2005-07-22)
    Dilated cardiomyopathy (DCM) or round heart disease is a muscle disease of the heart which is characterized by ventricular dilatation and abnormal systolic and diastolic left ventricular function. In animals, including turkeys and humans, DCM is the major cause of morbidity and mortality which results from heart failure. In the turkey, DCM can be idiopathic or induced. Since idiopathic or spontaneous DCM occurs in about 1-4% of normal turkeys, it is of significant concern to the poultry industry. In this study, it was proposed that the incidence and severity of DCM in the turkey may have a genetic basis. To test this hypothesis, I investigated differences in the incidence and severity of DCM in five domesticated turkey varieties including Blue Slate (BS), Bourbon Red (BR), Narragansett (N), Royal Palm (RP) and Spanish Black (SB). Preliminary investigations tested the reliability of echocardiography (ECHO) as a non-invasive and non-destructive technique for diagnosing DCM in a large number of birds from hatch to four weeks-of-age. One-day-old poults for both the preliminary and hypothesis testing investigations were obtained from Privett Hatcheries (Portales, New Mexico). The birds were raised under standard management conditions. In the preliminary investigation and to test my hypothesis, DCM was induced by feeding birds ad libitum standard diets containing 700 parts per million furazolidone. Results of the preliminary investigations showed that left ventricular end-diastolic dimension (LVEDD) and left ventricular end-systolic dimension (LVESD) were the most consistent ECHO indicators of DCM from hatch to 4 weeks-of-age. Variety differences in response to furazolidone were evaluated using these parameters as well as percent mortality. At 9 days-of-age, differences between control and treatment birds for percent mortality and LVESD were significant in the RP variety only but significant for LVEDD in RP and SB. At 29 and 33 days-of-age, all the pair-wise comparisons between control and treatment birds were significant for both LVEDD and LVESD. On average, the BR variety had the smallest dilatation of the heart and lowest mortality at 33 days-of-age when compared to other varieties. The results described in this thesis show, for the first time, variety differences in the turkey's response to diets containing furazolidone. They provide strong evidence that, like previous reports for idiopathic DCM, an animal's response to Fz-induced DCM has a strong genetic component.
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    The genetic ancestry of American Creole cattle inferred from uniparental and autosomal genetic markers
    Ginja, Catarina; Gama, Luis Telo; Cortes, Oscar; Martin Burriel, Inmaculada; Luis Vega-Pla, Jose; Penedo, Cecilia; Sponenberg, D. Phillip; Canon, Javier; Sanz, Arianne; do Egito, Andrea Alves; Angela Alvarez, Luz; Giovambattista, Guillermo; Agha, Saif; Rogberg-Munoz, Andres; Cassiano Lara, Maria Aparecida; Delgado Bermejo, Juan Vicente; Martínez, Amparo; Afonso, Sonia; Aguirre, Lenin; Armstrong, Eileen; Camacho Vallejo, María Esperanza; Canales, Amado; Cassama, Bernardo; Contreras, Gloria; Moras Cordeiro, J. M.; Dunner, Susana; Elbeltagy, Ahmed; Soares Fioravanti, Maria Clorinda; Gomez Carpio, Mayra; Gomez, Mariano; Hernandez, Antonio; Hernandez, Darwin; Juliano, Raquel Soares; Landi, Vincenzo; Marques, Ribamar; Martinez, Ruben D.; Roberto Martinez, O.; Melucci, Lilia; Molina Flores, Baldomero; Mujica, Fernando; Pares i Casanova, Pere-Miquel; Quiroz, Jorge; Rodellar, Clementina; Tjon, Gerald; Adebambo, Tumininu; Uffo, Odalys; Cesar Vargas, Julio; Villalobos, Axel; Zaragoza, Pilar (Springer Nature, 2019-08-07)
    Cattle imported from the Iberian Peninsula spread throughout America in the early years of discovery and colonization to originate Creole breeds, which adapted to a wide diversity of environments and later received influences from other origins, including zebu cattle in more recent years. We analyzed uniparental genetic markers and autosomal microsatellites in DNA samples from 114 cattle breeds distributed worldwide, including 40 Creole breeds representing the whole American continent, and samples from the Iberian Peninsula, British islands, Continental Europe, Africa and American zebu. We show that Creole breeds differ considerably from each other, and most have their own identity or group with others from neighboring regions. Results with mtDNA indicate that T1c-lineages are rare in Iberia but common in Africa and are well represented in Creoles from Brazil and Colombia, lending support to a direct African influence on Creoles. This is reinforced by the sharing of a unique Y-haplotype between cattle from Mozambique and Creoles from Argentina. Autosomal microsatellites indicate that Creoles occupy an intermediate position between African and European breeds, and some Creoles show a clear Iberian signature. Our results confirm the mixed ancestry of American Creole cattle and the role that African cattle have played in their development.
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    Genetic variation of susceptibility to fescue toxicosis in cattle
    Gould, Lowell (Virginia Tech, 1992-07-14)
    Fifteen calves of two sires were fed endophyte infected (E + ) fescue seed to quantify differences in susceptibility to fescue toxicosis among sire groups. One of the sires, a Polled Hereford, had a commercial reputation of producing calves with less severe symptoms of tall fescue toxicosis than their contemporaries. The control sire, also a Polled Hereford, had unknown merit with regard to offspring susceptibility to fescue toxicosis. The study was divided into five phases, two in which endophyte-infected fescue seed was included in the diet (phases two and four) and three in which fescue seed was not in the diet (phases one, three, and five). All calves were treated similarly and given the same opportunities for water, shade and socialization. Susceptibility to fescue toxicosis was measured by appetite (amount of feed consumed per day per unit of metabolic body weight), serum prolactin, cholesterol, and alkaline phosphatase concentrations, ability to maintain homeotherny (afternoon minus morning rectal temperature), ability to dissipate core body heat (afternoon minus morning surface temperature), and heat-transfer inefficiency (afternoon rectal minus afternoon surface temperatures). Appetite was decreased by the E + fescue seed, but the sire groups did not differ in their response to or recovery from E + fescue seed. The E + fescue seed caused prolactin to decrease (P < .0001) but the sire groups did not differ in their responses to addition or withdrawal of E + fescue seed from the diet. Cholesterol levels were lower overall (P < .001) when the E+ fescue seed was fed, and the sire groups recovered from the fescue toxicity at different rates (P < .001). Alkaline phosphatase concentrations were lower during the phases when E + fescue seed was fed; calves from the control sire were less resistant to the E + fescue seed effects than the Missouri calves (P < .0001). Ability to maintain homeothermy was reduced by the E + fescue seed, but the reduction did not differ between sire groups. Surface temperature changes and heat-transfer inefficiencies were not indicative of fescue toxicosis in this study. It was concluded that serum cholesterol and serum alkaline phosphatase concentrations may be accurate indicators of differences among paternal half sib groups of cattle in susceptibility to fescue toxicosis.
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    Investigation into the use of an epidural morphine sulfate and detomidine hydrochloride combination in horses: Part 1: efficacy in alleviation of hindlimb pain, Part 2: long-term systemic and local effects
    Sysel, Annette M. (Virginia Tech, 1996)
    In Part 1, amphotericin B-induced synovitis of the left tarsocrural joint was used to create hindlimb lameness in 11 horses. Caudal epidural catheters were placed and advanced to the lumbosacral region. Baseline heart and respiratory rates were recorded and horses were videotaped at a walk and trot. Treated horses received 0.2 mg/kg morphine sulfate and 30 ug/kg detomidine hydrochloride through the epidural catheter; control horses received an equivalent volume of physiologic saline solution through the catheter. At hourly intervals after epidural injection for a total of 6 hours, heart and respiratory rates were recorded and horses were videotaped walking and trotting. At the end of the observation period, video recordings were scrambled onto a master videotape. Lamenesses were scored by 3 investigators. Lameness grades, heart rates and respiratory rates were compared. There was a significant decrease in lameness grades after treatment with epidural morphine and detomidine. Initially, heart rates significantly increased in control horses and decreased in treated horses. A similar trend occurred for respiratory rates. In Part 2, caudal epidural catheters were used to administer injections to 10 horses every 12 hours for 14 days. Treated horses received 0.2 nlg/kg morphine sulfate and 30 ug/kg detomidine hydrochloride, and control horses received an equivalent volume of physiologic saline solution. Body weights were recorded on days 1 and 14. Rectal temperature, heart rate, respiratory rate and gastrointestinal motility were recorded twice daily, and daily hay and water consumption was measured. Horses were euthanatized day 15. Atlanto-occipital cerebrospinal fluid samples were submitted for bacteriologic culture and determination of white and red blood cell counts and protein and glucose concentrations. Post mortem examinations were performed and representative samples of the spinal cord and surrounding tissues were taken from cervicothoracic, thoracolumbar, lumbosacral, sacral and catheter entry point regions. Spinal tissue segments from these regions were graded for histologic degree of inflammation and fibrosis. Cerebrospinal fluid values and spinal tissue segment inflammation and fibrosis grades were compared between control and treated horses, and between all 10 catheterized study horses and 6 uncatheterized horses. No problems were encountered with epidural catheter maintenance or injection. No significant difference was identified in body weight change, daily variables or hay and water consumption between control and treated horses. All cerebrospinal fluid cultures were negative for growth. No significant difference in cerebrospinal fluid values or spinal tissue inflammation or fibrosis grades for any segment was demonstrated between control and treated horses. However, when compared to uncatheterized horses, cerebrospinal fluid red blood cell counts were marginally higher and protein concentrations were significantly higher in catheterized horses. As well, lumbosacral and sacral spinal tissue segment inflammation grades and sacral segment fibrosis grades were significantly higher in catheterized compared to uncatheterized horses. Results of these studies indicate that an epidural combination of morphine and detomidine provides profound hindlimb analgesia in horses and is not associated with apparent adverse systemic effects. Localized epidural inflammation and fibrosis appear to be catheter-related.
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    On the origins of American Criollo pigs: A common genetic background with a lasting Iberian signature
    Revidatti, Maria Antonia; Gama, Luis Telo; Martin Burriel, Inmaculada; Gardyn, Oscar Cortes; Cappello Villada, Juan Sebastian; Carolino, Maria Ines; Canon, Francisco Javier; Ginja, Catarina; Sponenberg, D. Phillip; Vicente, Antonio P.; Zaragoza, Pilar; Delgado Bermejo, Juan Vicente; Martínez, Amparo (2021-05-20)
    American Criollo pigs are thought to descend mainly from those imported from the Iberian Peninsula starting in the late 15th century. Criollo pigs subsequently expanded throughout the Americas, adapting to very diverse environments, and possibly receiving influences from other origins. With the intensification of agriculture in the mid-20th century, cosmopolitan breeds largely replaced Criollo pigs, and the few remaining are mostly maintained by rural communities in marginal areas where they still play an important socio-economic and cultural role. In this study, we used 24 microsatellite markers in samples from 1715 pigs representing 46 breeds with worldwide distribution, including 17 American Criollo breeds, with the major focus of investigating their genetic diversity, structure and breed relationships. We also included representatives of the Iberian, Local British, Hungarian, Chinese and Commercial breeds, as well as Wild Boar, in order to investigate their possible influence in the genetic composition of Criollos. Our results show that, when compared with the other breeds, Criollo pigs present higher levels of genetic diversity, both in terms of allelic diversity and expected heterozygosity. The various analyses indicate that breed differentiation overall explains nearly 21% of the total genetic diversity. Criollo breeds showed their own identity and shared a common genetic background, tending to cluster together in various analyses, even though they differ from each other. A close relationship of Criollos with Iberian breeds was revealed by all the different analyses, and the contribution of Iberian breeds, particularly of the Celtic breeds, is still present in various Criollo breeds. No influence of Chinese breeds was detected on Criollos, but a few were influenced by Commercial breeds or by wild pigs. Our results confirm the uniqueness of American Criollo pigs and the role that Iberian breeds have played in their development.
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    Prostaglandins Modulate the Uterine Response to Infectious Bacteria in Postpartum and Estrous Cyclic Ewes
    Seals, Richard (Virginia Tech, 2000-04-21)
    The uterine immune system is down regulated when exposed to P4. Five experiments were conducted to determine the in vivo and in vitro role(s) of P4 and prostaglandins in immune function(s) associated with uterine infections. In Exp. 1, postpartum ewes (d 0 = parturition) were supplemented with either OIL or P4 (beginning on d 10) or were SHAM or OVEX. Vena caval blood and lymphocytes were collected on d 14, and 16 to 19. All ewes received intrauterine inoculations of bacteria on d 15 and uteri were collected on d 20. Ewes receiving P4 developed infections. Lymphocytes were incubated with mitogens, PGE2, indomethacin (INDO; a PG synthesis inhibitor) or both in a 3 x 2 x 2 arrangement. Concanavalin A-stimulated blastogenesis in P4-OVEX ewes and PGE2 and PGE2+INDO treated lymphocytes was inhibited (P < .05). Cyclic ewes in their follicular or luteal phase received either intrauterine inoculations of saline or bacteria, vena caval blood was collected for 3 d, and uteri were collected. Lymphocytes were incubated with mitogens, PGE2 (Exp. 2), PGF2alpha (Exp. 3) and(or) INDO in a 3 x 2 x 2 arrangement. Only luteal phase ewes that received bacteria developed infections. In Exp. 2, Con A- and LPS-stimulated blastogenesis was greater for luteal than for follicular phase ewes. T lymphocyte proliferation was inhibited in ewes inoculated with bacteria. T lymphocyte proliferation tended to be higher (P = .09) when incubated with INDO. In Exp. 3, T lymphocyte proliferation in response to PGF2alpha was greater for follicular than for luteal phase ewes. Neutrophils were lower in ewes inoculted with bacteria. In Exp. 4 and 5, uteri of luteal-phase (d 6) ewes were inoculated with bacteria. Ewes received either 15 mg of Lutalyse or saline on d 9, and uteri were collected on d 11. Lutalyse reduced P4, tended to decrease neutrophils, allowed ewes to clear infections, and had no effect on blastogenesis. Methods for modulating uterine prostaglandins seem to reduce susceptibility to uterine infections.
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    Role of CD44 in Immune Functions and Endothelial Cell Injury
    Rafi-Janajreh, Asimah (Virginia Tech, 1998-09-17)
    In addition to the antigen-specific receptors, the T and B cells also express a variety of adhesion molecules, which are known to participate in cell-cell interaction, migration, homing and signal transduction. CD44 is a widely distributed cell surface glycoprotein whose principal ligand has been identified as hyaluronic acid (HA), a major component of the extracellular matrix. In the current study, we investigated whether HA or mAbs against CD44 would induce a proliferative response in mouse lymphocytes. Spleen cells from normal and nude but not severe combined immunodeficient mice, exhibited strong proliferative responsiveness to stimulation with soluble HA or anti-CD44 mAbs. Furthermore, purified B cells but not T cells were found to respond to HA. These data demonstrated that interaction between HA and CD44 can regulate murine B cell effector functions and that such interactions may play a critical role during normal or autoimmune responsiveness of B cells. Endothelial cell injury resulting in vascular leak syndrome (VLS) is one of the most widely noted phenomenons in a variety of clinical diseases, however, the underlying reason for which remains unclear. We used interleukin-2 induced VLS as a model to investigate the role of cytolytic lymphocytes in the direct cytotoxicity of endothelial cells. BL/6 wild-type mice developed significant VLS in the lungs, liver and spleen following IL-2 administration. Interestingly, perforin-knockout mice exhibited marked decrease in VLS in all three organs tested. Also, FasL-defective (gld) mice and Fas-deficient (lpr) mice exhibited decreased VLS in the liver and spleen, but not in the lungs. These results demonstrated for the first time that perforin and FasL may actively participate in endothelial cell injury and induction of VLS in a variety of organs. Inasmuch as, CD44 also plays a major role in the lymphocyte adhesion to the endothelial cells, we used CD44-knockout mice and observed that such mice exhibited markedly diminished VLS following IL-2-treatment. Our data also suggested that blocking CD44 helps in reducing the IL-2-induced VLS and therefore such an approach may serve as a useful tool to prevent endothelial cell damage seen in a variety of clinical disorders.
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    Tocopherol induced angiogenesis in placental vascular network in late pregnant ewes
    Kasimanickam, Ramanathan K.; Kasimanickam, Vanmathy R.; Rodriguez, Jacobo S.; Pelzer, Kevin D.; Sponenberg, D. Phillip; Thatcher, Craig D. (2010-07-12)
    Background Tocopherols have biphasic, proangiogenic and antiangiogenic therapeutic effects. The objective of this clinical trial was to clarify tocopherol's placental angiogenic potential in late pregnant ewes following oral supplementation. Methods Eighteen pregnant ewes during late gestation were selected for this study. Ewes were given oral supplementation of 500 mg of alpha-tocopherol (aT; N = 6) or 1000 mg of gamma-tocopherol (gT; N = 7) or placebo (CON; N = 5) once daily from 107 to 137 days post breeding. Serum was obtained at weekly intervals and tissue samples were obtained at the end of supplementation to: 1) evaluate tocopherol concentrations in serum, uterus and placentome; 2) evaluate relative mRNA expressions of Vascular Endothelial Growth Factor (VEGF), Placental Growth Factor (PlGF), endothelial Nitric Oxide Synthase (eNOS) and Hypoxia Inducible Factors (HIF) in uterus, caruncle and cotyledon; 3) analyze the morphometry of the placental vascular network. Results Supplementation of aT or gT resulted in increased concentrations in serum, placentome and uterus compared to control (P < 0.05). In aT group, mRNA expressions of PlGF, eNOS and HIF-1α in cotyledon were greater than the CON group. In gT group, mRNA expressions of VEGF, eNOS, HIF-1 alpha and HIF-2 alpha in caruncle and uterus, and HIF-1α in cotyledon, were greater than the CON group. Morphometry analysis revealed increased angiogenesis in the supplemented groups. Conclusion Daily oral supplementation of aT or gT increased angiogenesis in the placental vascular network in pregnant ewes during late gestation. Increase in placental angiogenesis may provide nutrients required for the development and growth of fetus during late pregnancy.
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    Tracing Worldwide Turkey Genetic Diversity Using D-loop Sequence Mitochondrial DNA Analysis
    Canales Vergara, Amado Manuel; Landi, Vincenzo; Delgado Bermejo, Juan Vicente; Martínez, Amparo; Cervantes Acosta, Patricia; Pons Barro, Águeda; Bigi, Daniele; Sponenberg, D. Phillip; Helal, Mostafa; Hossein Banabazi, Mohammad; Camacho Vallejo, María Esperanza (MDPI, 2019-11-01)
    According to recent archeological evidence, turkey (Meleagris gallopavo gallopavo) domestication may have occurred in Mexico around 2000 years ago. However, little is known about the phylogenetic and genealogical background underlying domestic turkey populations. This study aimed to further understand the domestication process and identify inter- or intraspecific connections between turkey populations to determine their origins, trace their global expansion, and define the species’ genetic value. Ninety-three domestic turkeys (local breeds) were sampled from populations in Brazil, Mexico, USA, Spain, Italy, Iran, and Egypt. Publicly available sequences from previous studies were also included. Standard mitochondrial DNA, genetic diversity, and haplotype network analyses were performed. Seventy-six polymorphic sites were identified. Turkeys from Mexico showed the greatest number of polymorphic sites (40), while turkeys from Italy and Brazil reported only one site each. Nucleotide diversity was also highest in Mexico and the USA (π = 0.0175 and 0.0102, respectively) and lowest in Brazil and Italy. Of the six major haplogroups defined, the Mexican and USA populations appeared to have remained more stable and diverse than the other populations. This may be due to conservative husbandry policies in the rural areas of other populations, which have prevented the introduction of commercial turkey lines.