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Analysis of mechanisms regulating the expression of parental alleles at the GPD locus in mule erythrocytes.

Erythrocyte glucose-6-phosphate dehydrogenase (G6PD) was examined by 13% starch gel electrophoresis in 74 mules (42 females and 32 males), 35 donkeys, and ten horses. The quantitative expression of the parental alleles at the Gpd locus varies greatly in female mules from the hemizygous expression of the maternal allele to that of the paternal. The data obtained indicate that the X chromosomes are randomly inactivated in females mules. No selective advantage of a cell population with a maternally (or paternally) derived X active was found in female mule erythrocytes. It is suggested that the phenotypic variability in the expression of the parental Gpd alleles is related to the random proportions established between cells having either a maternal or paternal X active in an initiator (stem) cell group giving rise to erythroid tissue. Initiator cell numbers estimated for erythroid tissue (six or seven) are close to those reported for human females and intergeneric fox hybrids. These numbers may vary depending on the duration of the time of determination and the division rate of initiator cells at determination.

Alleles↗

Merogony and gametogony of Eimeria mccordocki (Protozoa-Eimeriidae) in the mule deer, Odocoileus h. hemionus.

Eimeria mccordocki and E. madisonensis oocysts were isolated from feces of 21 of 40 captive mule deer in Fort Collins, Colorado. The two species were separated from each other by infecting one mule deer fawn, and the life cycle of E. mccordocki was studied for the first time. Four to six-weeks-old mule deer fawns were inoculated orally with E. mccordocki and killed 9, 13 and 15 days after infection. Asexual and sexual stages of life cycle developed in the ileum of mule deer, only in the surface epithelial cells of the villi. The asexual stages consisted of two generations of meronts.

Animals↗

Molecular cloning and sequence analysis of the cDNA encoding thyroid-stimulating hormone beta-subunit of common duck and mule duck pituitaries: in vitro regulation of steady-state TSHbeta mRNA level.

For better understanding of phylogenetic diversity and evolution of pituitary thyroid-stimulating hormone (TSH) in birds, we have cloned the cDNAs encoding TSH beta subunit (TSHbeta), by reverse transcription-polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE) from two species of domestic ducks, common duck (Tsaiya duck and Pekin duck) (Anas platyrhynchos domesticus) and mule duck (hybrid of male muscovy duck Cairina moschata and female A. platyrhynchos domesticus). The nucleotide sequences of isolated TSHbeta cDNAs of the two species of ducks are identical, with each including 66 bp of 5'-untranslated region (UTR), 402 bp of coding region, and 82 bp 3'-UTR followed by 18 bp poly A tract. The deduced TSHbeta subunit of the ducks contains 134 amino acids consisting of a putative signal peptide of 19 amino acids and a putative mature protein of 115 amino acids. However, the TSHbetas of common duck and mule duck differ from the TSHbeta of muscovy duck in one amino acid at position 97 of the mature protein: isoleucine for common duck and mule duck, and valine for muscovy duck. Our findings thus demonstrate that inter-genus variation of TSHbeta exists in Family Anatidae, and that TSHbeta gene in the mule duck is preferentially transcribed from the maternal genome rather than from the paternal genome. TSHbeta mRNA expressions were investigated by culturing common duck pituitaries with various doses of hormones. Thyrotropin-releasing hormone (TRH) stimulated, while thyroid hormones, triiodothyronine (T(3)) and thyroxine (T(4)), inhibited the TSHbeta mRNA levels, in dose-related manners. The findings thus support that the mode of regulation of TSH gene expression in hypothalamo-pituitary-thyroid axis in birds is similar to that in mammals. Cortisol and corticosterone decreased the steady-state TSHbeta mRNA levels at the pituitary level, in a dose-related manner, the first-time demonstration in vertebrates. The results may suggest that glucocorticoids exert suppressive action directly at pituitary level in modulation of steady-state TSHbeta mRNA level.

Amino Acid Sequence↗

Assessment of the welfare of working horses, mules and donkeys, using health and behaviour parameters.

Working animals provide an essential transport resource in developing countries worldwide. Many of these animals are owned by poor people and work in harsh environments, so their welfare is a cause for concern. A protocol was developed to assess the welfare of working horses, mules and donkeys in urban and peri-urban areas, using direct observation of health and behaviour parameters. In this study, 4903 animals used for draught, pack and ridden work in Afghanistan, Egypt, India, Jordan and Pakistan were assessed between December 2002 and April 2003. The data showed that donkeys were more likely than mules or horses to demonstrate avoidance or aggressive behaviour towards an observer, while horses were most likely to make a friendly approach. Fewer than 8% of working equines had abnormal mucous membranes, ectoparasites or poor coat condition. Body lesions occurred predominantly in the areas of the breast/shoulder, withers and girth in all three species, with mules having the highest prevalence of lesions in these areas (22.5, 21.3 and 28.4%, respectively). Among horses and donkeys, the prevalence of these lesions was influenced by the type of work carried out. Lesions on the head, neck, ribs, flank and tail base were seen in less than 10% of animals. Across all three species approximately 70% of animals were thin, having a body condition score (BCS) of 2 or less on a scale of 1-5 (1, very thin; 5, very fat) and more horses were in very thin condition (BCS 1) than mules or donkeys. Over 75% of animals demonstrated limb deformities and abnormalities of gait. The results of this study are being used as the initial stage of a long-term strategy to inform priorities for welfare interventions in working equines and to establish a welfare benchmark. Subsequent stages will rank the welfare concerns identified, assess the contributing risk factors and implement specific interventions to address these risks. Following intervention, success in improving welfare will be measured by repetition of this protocol and comparison with the benchmark.

Animal Welfare↗

Mutator transposon in maize and MULEs in the plant genome.

Mutator (Mu) is by far the most mutagenic plant transposon. The high frequency of transposition and the tendency to insert into low copy sequences for such transposon have made it the primary means by which genes are mutagenized in maize (Zea mays L.). Mus like elements (MULEs) are widespread among angiosperms and multiple-diverged functional variants can be present in a single genome. MULEs often capture genetic sequences. These Pack-MuLEs can mobilize thousands of gene fragments, which may have had a significant impact on host genome evolution. There is also evidence that MULEs can move between reproductively isolated species. Here we present an overview of the discovery, features and utility of Mu transposon. Classification of Mu elements and future directions of related research are also discussed. Understanding Mu will help us elucidate the dynamic genome.

DNA Transposable Elements↗

Variability of plasma melatonin level in pony mares (Equus caballus), comparison with the hybrid: mules and with jennies (Equus asinus).

In long-day breeders like horses, the length of nocturnal melatonin secretion is the main messenger of photoperiod. Previous studies have shown that the nocturnal jugular melatonin concentration is lower in horses, than in mules but is unknown in donkeys. The aim of this study was to estimate the inter-animal variability of plasma melatonin concentration in domestic mares and to compare this concentration with those observed in domestic jennies and in their hybrid mules. In the autumn, blood samples were collected at 22 h, 23 h, 0 h and 1 h during 2 nights at 3 weeks intervals, in 110 pony mares, 10 jennies and 6 mules maintained under natural photoperiod. Melatonin was assayed by a validated RIA method. The statistical analysis of the measures was done with a specific unbalanced analysis of variance model. The effect of species and individuals (nested under species) was highly significant. The mean melatonin concentration was 24 pg.mL(-1) in mares and was significantly lower than in jennies and in mules which were 90 pg.mL(-1) and 169 pg.mL(-1) respectively. The melatonin plasma concentration was higher in jennies than in mares. These results suggest that the melatonin concentration is genetically determined.

Animals↗

Mitochondrial DNA analysis of hybridization between sympatric white-tailed deer and mule deer in west Texas.

Sympatric populations of white-tailed deer and mule deer (Odocoileus virginianus and Odocoileus hemionus, respectively) on a west Texas ranch share a common mitochondrial DNA restriction map genotype. Phylogenetic analysis indicates that this genotype is more characteristic of O. virginianus than of O. hemionus. The genotype of west Texas deer differs from that of O. virginianus from South Carolina by five mutational events (1.3% sequence divergence), whereas it differs from that of O. hemionus from California by 17 events (5.5% divergence). We suggest that interspecies hybridization has occurred, primarily between mule deer bucks and white-tailed deer does, with preferential absorption of hybrid offspring into the mule deer gene pool. Introgressive hybridization may be involved in ongoing displacement of mule deer by white-tailed deer in west Texas.

Animals↗

Spatial heterogeneity of mitochondrial DNA and allozymes among populations of white-tailed deer and mule deer.

A white-tailed deer (Odocoileus virginianus) population in northeastern Minnesota and a mule deer (O. hemionus) population in the Bridger Mountains Montana, have previously been shown to be spatially subdivided into contiguous subpopulations. We assessed the degree of genetic differentiation among subpopulations and tested the hypothesis that differentiation will be greater for mitochondrial DNA (mtDNA) than for nuclear-encoded allozymes. Differentiation of the white-tailed deer subpopulations was significant for two allozyme loci but not for mtDNA, and the overall degree of differentiation was low. Gene flow, recent founding of the subpopulations, and polygamous breeding structure may all have contributed to this pattern. Greater differentiation was evident among disjunct populations than between the contiguous subpopulations of white-tailed deer. The contiguous mule deer subpopulations were significantly differentiated for mtDNA and one allozyme locus. Differentiation was greater for mtDNA than for allozymes. These results are consistent with demographic data that indicate mule deer males disperse more than do females. Disjunct mule deer populations may be similar or dramatically different in mtDNA haplotype frequencies that do not necessarily vary with geographic distance. Current and historical gene flow and breeding structure will influence population genetic patterns.

Animals↗

The evolutionary fate of MULE-mediated duplications of host gene fragments in rice.

DNA transposons are known to frequently capture duplicated fragments of host genes. The evolutionary impact of this phenomenon depends on how frequently the fragments retain protein-coding function as opposed to becoming pseudogenes. Gene fragment duplication by Mutator-like elements (MULEs) has previously been documented in maize, Arabidopsis, and rice. Here we present a rigorous genome-wide analysis of MULEs in the model plant Oryza sativa (domesticated rice). We identify 8274 MULEs with intact termini and target-site duplications (TSDs) and show that 1337 of them contain duplicated host gene fragments. Through a detailed examination of the 5% of duplicated gene fragments that are transcribed, we demonstrate that virtually all cases contain pseudogenic features such as fragmented conserved protein domains, frameshifts, and premature stop codons. In addition, we show that the distribution of the ratio of nonsynonymous to synonymous amino acid substitution rates for the duplications agrees with the expected distribution for pseudogenes. We conclude that MULE-mediated host gene duplication results in the formation of pseudogenes, not novel functional protein-coding genes; however, the transcribed duplications possess characteristics consistent with a potential role in the regulation of host gene expression.

DNA Transposable Elements↗

Circadian and seasonal rhythms of melatonin production in mules (Equus asinus x Equus caballus).

The present study describes the patterns of melatonin production in the mule (Equus asinus x Equus caballus). Blood was sampled hourly for 24 h from eight mule mares in spring and fall. The data obtained show the presence of a circadian rhythm of production of melatonin, with highest values during the dark phase both in spring and fall. In fall the nightly rise of melatonin production begins earlier in the dark phase and reaches higher quantitative levels than in spring. The morning decline of melatonin production is similar in the two seasons. Maximal levels of nightly melatonin production in the mule are about 10 x higher than those described in the horse. The results reported here indicate the persistence of brain structures able to receive and transduce environmental signals in the mule, a genetically sterile mammalian hybrid.

Animals↗

Validation of monoclonal antibody F99/97.6.1 for immunohistochemical staining of brain and tonsil in mule deer (Odocoileus hemionus) with chronic wasting disease.

A new monoclonal antibody (MAb), F99/97.6.1, that has been used to demonstrate scrapie-associated prion protein PrP(Sc) in brain and lymphoid tissues of domestic sheep with scrapie was used in an immunohistochemistry assay for diagnosis of chronic wasting disease (CWD) in mule deer (Odocoileus hemionus). The MAb F99/97.6.1 immunohistochemistry assay was evaluated in brain and tonsil tissue from 100 mule deer that had spongiform encephalopathy compatible with CWD and from 1,050 mule deer outside the CWD-endemic area. This MAb demonstrated abnormal protease-resistant prion protein (PrP(res)) in brains of all of the 100 mule deer and in 99 of the 100 tonsil samples. No immunostaining was seen in samples collected from deer outside the endemic area. MAb F99/97.6.1 demonstrated excellent properties for detection of PrP(res) in fresh, frozen, or mildly to moderately autolytic samples of brain and tonsil. This immunohistochemistry assay is a sensitive, specific, readily standardized diagnostic test for CWD in deer.

Animals↗

Evaluation of microchip migration in horses, donkeys, and mules.

OBJECTIVE: To determine whether microchips used for identification migrate after implantation in horses, donkeys, and mules. DESIGN: Prospective study. ANIMALS: 53 horses, donkeys, and mules. PROCEDURE: Twenty horses that had had microchips implanted in the nuchal ligament at a veterinary teaching hospital from 1996 through early 2000 were included (group 1), and the poll-to-withers distance and location of the microchip were determined, measured, and recorded. Additionally, the poll-to-withers distance was measured in 16 horses, 12 donkeys, and 5 mules (group 2), and microchips were implanted in the nuchal ligament on the left side of the neck. Forty-two to 67 days after implantation, the location of the microchip was determined, measured, and recorded. RESULTS: Microchips implanted in the nuchal ligament < or = 4 years previously did not migrate. All microchips were detected with a multimode identification tag reader from the left side of the neck in the midcervical region, and microchips were located at the midpoint between the poll and withers for all 53 horses, donkeys, and mules. CONCLUSIONS AND CLINICAL RELEVANCE: Microchips implanted in the nuchal ligament < or = 4 years earlier did not migrate in horses. Microchips may be useful for identification in horses.

Animal Identification Systems↗

Elaeophorosis in Barbary sheep and mule deer from the Texas Panhandle.

Adult Elaeophora schneideri were recovered from the common carotid artery and its branches in 14 of 14 mule deer, Odocoileus hemionus, and 3 of 9 Barbary sheep or aoudads, Ammotragus lervia, from Palo Duro Canyon in the Texas Panhandle. Gross cutaneous lesions attributable to elaeophorosis in the Barbary sheep varied from small circumscribed scars up to 10 cm in diameter usually on the poll or orbital region to extensive proliferative irregular encrustations on the frontal, temporal and orbital regions, sometimes extending to the ears and muzzle. Individual lesions varied from slate-gray scarred areas to brown proliferative edematous and hyperemic encrustations, sometimes with depigmented pustules a few millimeters in diameter. Microscopic lesions ranged from granulation tissue to severe pyogranulomatous reactions with neutrophils, eosinophils, lymphocytes and plasma cells as the primary infiltration. Foreign body giant cells and/or microfilariae were not observed. Microscopic changes in the carotid arteries and their branches were limited to small villous projections on the intimal surface apparently resulting from medial hyperplasia. Cutaneous lesions attributable to elaeophorosis were not observed in mule deer. Histopathologic lesions in the carotid arteries of mule deer were similar to those observed in Barbary sheep. The comparative pathology of elaeophorosis in various hosts is reviewed and discussed in terms of its pathology in Barbary sheep. The potential ramifications of this infection on the expanding aoudad population in the southwestern United States require that elaeophorosis be considered in the management of this species, particularly in areas with sympatric mule deer populations.

Animals↗

Immobilization of mule deer with thiafentanil (A-3080) or thiafentanil plus xylazine.

We evaluated thiafentanil oxalate (A-3080) for the immobilization of mule deer (Odocoileus hemionus) under laboratory and field conditions. In a crossover experiment comparing recommended (0.1 mg/kg) and 2x recommended thiafentanil doses in captive deer, both produced rapid induction and immobilization. Mean induction was shorter (P = 0.013) for the 2x group (1.9 vs. 3 min); mean reversals for both groups were rapid (recommended = 0.9 min after naltrexone injection; 2x = 1 min) and did not differ (P = 0.29). Six free-ranging mule deer were immobilized with 7 mg thiafentanil and four with 10 mg; mean induction was 2.3 min for both groups (95% confidence interval [CI]: 7 mg, 1.2-3.4; 10 mg, 1.9-2.8), and mean reversal was <1 min for both groups. Of 165 free-ranging deer darted with various combinations of thiafentanil and xylazine, we successfully immobilized 148 (90%). Mean induction ranged from 2.1 to 4.9 min for different drug combinations. Reversals were not compared because naltrexone and yohimbine doses varied, but overall mean reversal was 1.9 min (95% CI, 1.7-2.1 min) after injection of naltrexone and yohimbine intravenously (i.v.); naltrexone:thiafentanil ratios ranging from 10:1 to 43:1 provided mean recoveries ranging from 1.5 to 2.3 min. All 25 deer fitted with radio collars were alive at 30 days postcapture. On the basis of overall reliability and effectiveness, drug volumes, and ease of handling drugged animals, we recommend using a combination of 10-12 mg thiafentanil (0.15-0.2 mg/kg) and 100 mg xylazine to immobilize mule deer; immobilization can be effectively reversed with 100 mg naltrexone or more and 15 mg yohimbine or more i.v. Where feasible, we also recommend the use of transmitter darts when immobilizing mule deer with opioids in order to maximize recovery of darted deer and to ensure that missed darts are found.

Adrenergic alpha-Agonists↗

Determining prevalence of bluetongue and epizootic hemorrhagic disease viruses in mule deer in Arizona (USA) using whole blood dried on paper strips compared to serum analyses.

We investigated the feasibility of using whole blood dried on paper strips as a means to collect antibody prevalence data for the epizootic hemorrhagic disease viruses (EHDV) and bluetongue viruses (BTV) from hunter-harvested male mule deer (Odocoileus hemionus) in October 2002 from Arizona, USA. We compared antibody prevalence estimates in mule deer from paired paper strip and serum samples. Prevalence data obtained from elution of dried blood on paper strips proved to be consistent with results from serum in 94% of the samples tested. The paper strip method allows easy collection of blood from dead animals, with a smaller amount of blood being needed for analyses. Also, samples do not need to be refrigerated before analyses. We also used serum samples to determine hemorrhagic disease (HD) serotype exposure status of mule deer harvested from 4 distinct areas in Arizona. Antibodies to BTV and EHDV were identified in 3 of the 4 areas, with positive results to EHDV-1, EHDV-2, BTV-10, and BTV-11 being most common. Many animals did not have antibodies against the BTV serotypes. Exposure varied geographically and potentially with elevation. Hemorrhagic disease viruses commonly infect Arizona mule deer, except on the Kaibab Plateau in northern Arizona.

Animals↗

Morphological examination of epididymal epithelium in the mule (E. hinnus) in comparison with parental species (E. asinus and E. caballus).

Following previous studies about the ultrastructure of male genital tract in parental species, a comparative study of epididymis of one of the possible hybrids, the mule, has been undertaken. Apart from small differences, general features of epididymal epithelium in the mule are similar to those of parental species. However, extension of our studies from the donkey to the horse to the hybrid permits a deeper insight into the morphology of this tract of excurrent duct. In the meantime, it is possible to evidence some features, sometimes shared with other species if taken separately, which in the whole characterize the epididymis in Equidae: the presence in principal cells of intranuclear inclusions and peculiar small granules in the basal cytoplasmic edge; the organization of groups of cells, likely to be principal ones, in such a way as to constitute intraepithelial crypts; a cumbersome presence of lipofuscinic matter all along the epithelium. Another interesting observation in the presence in the mule epididymis of well recognizable macrophages. All these data are discussed in comparison with parental species and with other species described in literature. Beyond any other consideration, it can be outlined that the complex morphology of the epithelium lining ductus epididymis in the mule is unaffected by the absence of spermatozoa, which are normally the target of the manifold functions of the epithelium itself.

Anatomy, Veterinary↗

Evaluation of free-roaming mule deer as carriers of anaplasmosis in an area of Idaho where bovine anaplasmosis is enzootic.

Samples of blood from 87 Rocky Mountain mule deer (Odocoileus hemionus hemionus) were inoculated into 3 susceptible splenectomized calves to determine the anaplasmosis carrier status of the deer. The deer were trapped along the Idaho-Utah state boundary, near Stone, Id. Acute anaplasmosis was induced in 2 of the 3 inoculated calves, and blood from the 3 calves caused anaplasmosis when inoculated into adult non-splenectomized cattle. Serum rapid card agglutination testing revealed 13 (14.9%) positive reactions among the 87 mule deer. When these seropositive samples were evaluated with the complement-fixation test, positive reactions were not found and only 3 (3.5%) serums gave suspect reactions. A serologic survey for anaplasmosis in 1,852 cattle that cohabit spring and summer rangeland areas with mule deer revealed 14.5% reactors by the serum rapid card agglutination test. The study area consisted of an ecosystem that supports cattle, free-roaming mule deer, and Dermacentor andersoni, and thus contained the elements necessary to effect interspecies transmission of Anaplasma marginale. The significance of wildlife reservoirs and their bearing on anaplasmosis control and eradication programs needs further investigation.

Anaplasmosis↗

Fine structure of mule testes: light and electron microscopy study.

Testes of 3 mules, 3 stallions, and 3 donkeys were studied by light and electron microscopic techniques. Mule testes were firm, with an average weight less than that of stallion or donkey testes. As expected, mule seminiferous tubules had few cell elements. There were 2 cell populations within the seminiferous tubules: (a) Only basal spermatogonia and Sertoli's cells in 70% of the tubules and, (b) the cells in (a) plus leptotene and pachytene primary spermatocytes in 30% of the tubules. Degenerative changes occurred at the pachytene stage and began with the appearance of nuclear vesicles and chromatin condensation. The last stages disclosed pyknotic nuclei and cytoplasmic vesicles. The interstitia were filled by connective tissue elements and few Leydig's cells. Interstitial cells had well-developed, smooth endoplasmic reticula and mitochondria, indicating a normal steroidogenic activity. Epididymides in the mule had morphologic features similar to those of the stallion or donkey, except for the presence of intertubular fibrosis. Degenerative changes in primary spermatocytes indicate metabolic alterations which could be explained by the blockage of meiosis.

Animals↗