Studies on blood serum beta-N-acetylglucosaminidases from several mammalian species--separation of different enzyme forms.

1. The beta-N-acetylglucosaminidases studied from peripheral blood sera have the following activities: 55.6 mU/ml in Bos taurus L. (bull, "Morucha" race), 31.3 mU/ml in Sus scropha var. domestica L. (pig) and 3.7 mU/ml in Capra hircus L. (goat). 2. The pH optima of the enzymes are 4.5 for Bos taurus L. and Sus scropha L., and 5.0 for Capra hircus L. 3. Two enzyme forms, A and B, have been separated from Bos taurus L. and Capra hircus L. sera by DEAE-cellulose chromatography; and three forms, A, B and S, from Sus scropha L. serum. 4. The B forms are more heat-stable than the A and the S forms. The S form has intermediate heat-stability between the A and the B forms.     

Isolation and chemical composition of the leucocytes from donkey, horse, mule and pig.

1. Leucocytes from the blood of adult and young donkeys (Equus asinus L.), adult horses (Equus caballus L.), adult mules (Equus asinus x Equus caballus) and adult pigs (Sus scrofa L.) were obtained in a high degree of purity (99.9%) using Na2-EDTA-dextrans mixtures. 2. Sialic acids were released, purified, identified and determined from both non-delipided and delipided leucocytes. 3. N-glycolylneuraminic was the predominant sialic acid. N-acetylneuraminic acid and N,O-diacetyl-neuraminic acid were also found in all materials. Except in pig, other unidentified sialic acid(s) were also detected. 4. The concentration of total sialic acids (microgram/mg protein) is different according to the species, and in donkey species according to the age. 5. Galactose, glucose, mannose, fucose and (in a less amount) ribose were determined. Their total content is about 2-3-fold that of hexosamines. 6. There is a higher cholesterol content in adult donkey leucocytes than in those of young ones. 7. Total lipids, cholesterol or phospholipid contents are similar among the leucocytes of the above-mentioned species. 8. The similarities are marked in the electrophoresis patterns of proteins and glycoproteins for the donkey, mule and horse samples. 9. The molecular weights for leucocytes proteins were estimated as ranging between 230,000 and 20,000; and for the main protein bands, between 120,000 and 22,000.     

Comparative studies on six blood serum glycosidases from several mammalian species.

1. Peripheral blood serum alpha-D-galactosidase, beta-D-galactosidase, beta-D-glucosidase, alpha-D-mannosidase, beta-D-xylosidase and beta-D-glucuronidase have been studied with a comparative point of view from several mammalian species: Bos taurus L. (bull), Capra hircus L. (goat), Sus scropha var. domestica L. (pig) and man. 2. Fluorimetric and spectrophotometric procedures were used for determination of enzyme activities and pH optima. 3. Glycosidase activity was generally higher with fluorescent substrates than with chromogenic substrates. 4. alpha-D-mannosidase was the most active with both fluorescent and chromogenic substrates. 5. All the studied enzymes had the same pH optimum (4.0) when the chromogenic substrates were used. 6. pH optima of these glycosidases ranged from 3.0 to 5.5 when the fluorescent substrates were used.     

Serologic surveillance for vesicular stomatitis virus on Ossabaw Island, Georgia

Seventeen species of mammals and seven species of birds from Ossabaw Island, Georgia, were tested for vesicular stomatitis (VS) neutralizing antibodies. Seropositive results were restricted to mammals with six of 17 species testing seropositive for VS (New Jersey type) neutralizing antibodies. Seropositive species included: raccoons (Procyon lotor), white-tailed deer (Odocoileus virginianus), feral swine (Sus scrofa), cattle (Bos taurus), horses (Equus caballus), and donkeys (Equus asinus). All tests for VS (Indiana type) were negative.     

Histomorphometric study of bone microstructure of primates and domestic animal with the goal of species identification with reference to the effects of domestication

Functional bone microstructure of long limb bones is a function of species-specific biomechanical properties such as locomotion and weight. Histomorphometry and statistics were used to identify various primate species (Hylobates moloch, Pongo satyrus borneensis, Pan tr. troglodytes, Gorilla g. gorilla, Homo sapiens), equid species (Equus caballus, Equus asinus, Equus mulus, Equus hemionus kulan, Equus ferus przewalskii) and also extinct horses e.g. iron age, medieval and neolithic forms on the microstructural level. Furthermore, bones from domesticated cattle, their Neolithic forms, pigs, sheep and goats (Bos taurus, Sus scrofa, Ovis aries, Capra hircus) were examined. Thin sections from proximal metacarpi or radii per species were taken in case of the domesticated animals and from distal humeri in case of the primates. Areas, perimeters, minimal and maximal axis of Haversian canals and secondary osteons were measured on digital images. Canonical discriminant analysis permits a differentiation of the species by these parameters of bone microstructure. Thus it is possible to distinguish between the different primate species, sheep and goats, horses, extinct horses, donkeys, mules and kulans on the microstructural level, however not between cattle and pig, E. f. przewalskii and Equus caballus, medieval and iron age horses. Neolithic cattle and horses do overlap, yet they are different from the modern forms.     

Comparative analysis of the peroxidase activity of myoglobins in mammals

Studies have been made on the peroxidase activity of metmyoglobins in animals from various ecological groups--the horse Equus caballus, cattle Bos taurus, beaver Castor fiber, otter Lutra lutra, mink Mustela vison and dog Canis familiaris. It was found that the level of this activity in diving animals depends on the duration of their diving, whereas in terrestrial species--on the strength of muscular contraction.     

Evolutionary movement of centromeres in horse, donkey, and zebra

Centromere repositioning (CR) is a recently discovered biological phenomenon consisting of the emergence of a new centromere along a chromosome and the inactivation of the old one. After a CR, the primary constriction and the centromeric function are localized in a new position while the order of physical markers on the chromosome remains unchanged. These events profoundly affect chromosomal architecture. Since horses, asses, and zebras, whose evolutionary divergence is relatively recent, show remarkable morphological similarity and capacity to interbreed despite their chromosomes differing considerably, we investigated the role of CR in the karyotype evolution of the genus Equus. Using appropriate panels of BAC clones in FISH experiments, we compared the centromere position and marker order arrangement among orthologous chromosomes of Burchelli's zebra (Equus burchelli), donkey (Equus asinus), and horse (Equus caballus). Surprisingly, at least eight CRs took place during the evolution of this genus. Even more surprisingly, five cases of CR have occurred in the donkey after its divergence from zebra, that is, in a very short evolutionary time (approximately 1 million years).These findings suggest that in some species the CR phenomenon could have played an important role in karyotype shaping, with potential consequences on population dynamics and speciation.     

The oxygen affinity of mammalian hemoglobins in the absence of 2,3-diphosphoglycerate in relation to body weight

We studied the oxygenation of mammalian hemoglobins: mouse (Mus musculus molossinus), rabbit (Oryctolagus cuniculus domesticus), Japanese monkey (Macaca fuscata), man (Homo sapiens), sheep (Ovis aries), llama (Lama glama), pig (Sus scrofa domesticus), cow (Bos taurus domesticus) and horse (Equus caballus), in the absence of 2,3-diphosphoglycerate (DPG) and compared their oxygen affinity in relation to the body weight. The negative correlation between body weight and the oxygen affinity of the whole blood, observed by Schmidt-Nielsen and Larimer (1958), was not observed in the absence of DPG. Our results indicated that an adaptive evolution proposed for hemoglobin in terms of its oxygen affinity vs body weight of the animal can only be appreciated with DPG.     

Variation in skin surface lipid composition among the Equidae

Skin surface lipids from Equus caballus, E. przewalskii, E. asinus, E. grevyi, E. hemionus onager and a mule (E. asinus/E. caballus) were analyzed in detail. In all species the surface lipid mixtures consisted of giant-ring lactones, cholesterol, cholesteryl esters and minor amounts of wax diesters. In E. caballus, the lactone hydroxyacids were entirely branched chained, while in E. asinus and E. grevyi they were almost exclusively straight chained. In E. przewalskii, the onager and the mule there were both straight and branched chain hydroxyacid lactones. These results are in harmony with published interpretations of the evolutionary relationships among Equus species.     

Characterization of enterococci populations in livestock manure using BIOLOG

The BIOLOG system was used to generate knowledge of enterococci populations found in fresh and dry manure of livestock (cattle (Bos taurus), horse (Equus caballus), and sheep (Ovis aires)). Six-hundred and forty Enterococcus isolates from the host sources were observed as a combined fresh manure unit and a combined dry manure unit, E. casseliflavus and E. mundtii were predominant in fresh manure (36% and 35%, respectively) as well as in dry manure (51% and 28%, respectively). The other species were found at a frequency of less than 10%. A chi-square test of the two most predominant Enterococcus sp. indicated that there were some significant differences among the frequency of E. casseliflavus and E. mundtii in cattle and sheep, but not horse. Despite these differences, these two species were overwhelmingly predominant among all three livestock sources.     

Numbers of fecal streptococci and Escherichia coli in fresh and dry cattle, horse, and sheep manure

Livestock are known contributors to stream pollution. Numbers of fecal streptococci and Escherichia coli in manure naturally deposited by livestock in the field are needed for activities related to bacterial source tracking and determining maximum daily bacterial loading of streams. We measured populations of fecal streptococci and E. coli in fresh and dry manure from cattle (Bos taurus L.), horses (Equus caballus L.), and sheep (Ovis aires L.) on farms in southern Idaho. Populations of indicator bacteria in dry manure were often as high as that in fresh manure from horse and sheep. There was a 2 log10 drop in the population of fecal coliform numbers in dry cattle manure from cattle in pastures but not from cattle in pens. Bacterial isolates used in source tracking should include isolates from both fresh and dry manure to better represent the bacterial source loading of streams.     

巫山蓝家寨遗址家畜的动物骨骼

2012年巫山蓝家寨遗址考古发掘中,出土了春秋时期的马(Equus caballus)、黄牛(Bos taurus)、山羊(Capra hircus)、鸡(Gallus gallus domesticus)、狗(Canis familiaris)、猪(Sus domestica)6种家畜的骨骼。这是三峡地区唯一一个在先秦时期六畜兼备的古文化遗址。遗址中的马、牛,是三峡地区同类家畜中时代最早的发现资料。三峡地区早期历史时期,人类肉食资源以野生动物为主,家畜动物骨骼发现材料较少。蓝家寨遗址出现种类较多的家畜动物种类,为全面分析和认识三峡地区经济形态发展过程、古居民行为模式和区域文化等,提供了重要资料。遗址中在春秋时期出现的家畜马和黄牛,与该时期各种区域文化在三峡地区的碰撞、交流、融合有关。     

Conserved restriction sites within the ribosomal RNA genes of vertebrates

We have mapped the cleavage sites of four restriction enzymes which recognize six-base sequences within the nuclear ribosomal (rRNA) genes of twelve vertebrates, including several placental mammals (Homo sapiens, man; Bos taurus, cow; Equus caballus, horse; Sus scofra, pig; Ovis aries, sheep; Rattus rattus, rat), a marsupial (Didelphis marsupialis, opossum), a bird (Gallus domesticus, chicken), an amphibian (Xenopus laevis), a reptile (Alligator mississipiensis), a bony fish (Cynoscion nebulosus, sea trout), and a cartilagenous fish (Carcharhinus species, requiem shark). These animals represent a span of approx. 400 million years of evolutionary divergence. Our data identify restriction sites in the rRNA genes which are highly conserved among higher vertebrates and therefore are likely to be in functionally important regions. Additionally, the restriction enzyme sites identified will be useful in cloning and sequencing the rRNA genes in any vertebrate. Finally, the consistent size and conserved sequence homology suggests that these rRNA gene segments will be useful as internal controls in hybridization experiments involving other genomic regions in vertebrates.     

Chromosomal assignment of six genes (EIF4G3, HSP90, RBBP6, IL8, TERT, and TERC) in four species of the genus Equus

We mapped six genes (EIF4G3, HSP90, RBBP6, IL8, TERT, and TERC) on the chromosomes of Equus caballus, Equus asinus, Equus grevyi, and Equus burchelli by fluorescence in situ hybridization. Our results add six type I markers to the cytogenetic map of these species and provide new information on the comparative genomics of the genus Equus.     

内蒙古白绒山羊4E-BP1基因cDNA克隆及组织表达特异性分析

旨在克隆内蒙古白绒山羊4E-BP1(真核细胞翻译起始因子4E结合蛋白1)基因并进行生物信息学及表达模式分析。根据已报道物种4E-BP1基因cDNA序列,用primer premier5软件设计引物,通过RT-PCR从绒山羊胎儿成纤维细胞总RNA中扩增出4E-BP1基因编码区cDNA序列,对目的片段进行测序及表达模式分析。克隆到的内蒙古白绒山羊4E-BP1基因cDNA全长357 bp,包含了完整的的ORF,编码118个氨基酸残基。核酸序列与牛、马、人、大鼠及小鼠的同源性分别为98%、90%、90%、88%和87%。4E-BP1基因在绒山羊脑、心脏、睾丸及胰腺组织中均有表达。     

Glycosylation in the near-term epitheliochorial placenta of the horse, donkey and camel: a comparative study of interbreeding and non-interbreeding species

Studies from this laboratory have shown great diversity in the glycosylation of tissues comprising the interhaemal barrier of species with different placental types. This diversity may be one of the factors preventing interbreeding between species. Glycan expression within the uterine epithelium and trophoblast of the interhaemal barrier was examined to test this proposition in three species with similar diffuse, microcotyledonary, epitheliochorial allantochorionic types of placenta: the horse (Equus caballus) and donkey (Equus asinus), which can interbreed with each other, and the camel (Camelus dromedarius), which cannot interbreed with either of the other two species. A panel of 14 lectins was used and it was found that glycosylation patterns were generally similar between placental tissues of the horse and donkey, except for the expression of non-bisected complex N-glycan and some sialic acids, whereas those of the camel showed striking differences in the binding of lectins to many structures carrying terminal residues of fucose, N-acetyl galactosamine and beta-galactose, as well as to complex N-glycans and sialic acids. These results are consistent with the proposition that interbreeding species carry similar glycans in tissues forming the interhaemal barrier whereas glycodiversity is one of the factors preventing implantation and subsequent placental development in interspecific hybrids.     

Comparative testis morphometry and seminiferous epithelium cycle length in donkeys and mules

The mule (Equus mulus mulus) is a sterile hybrid domestic animal that results from the breeding of a male donkey (Equus asinus) to a female horse (Equus caballus). Usually, spermatogenesis in mules does not advance beyond spermatocytes. In the present study, we performed a comparative and more accurate morphometric and functional investigation of the testis in donkeys and mules. Due to the smaller testis size, lower seminiferous tubule volume density, and fewer germ cells, the total length of seminiferous tubules in mules was significantly smaller than in donkeys. However, the percentage of seminiferous tubules containing germ cells (spermatogonia and spermatocytes) in mules was approximately 95%. The total number of Sertoli cells per testis observed in donkeys and mules was very similar. However, the total number of Leydig cells in mules was approximately 70% lower than in donkeys. At least in part, this difference was probably related to the lower number of germ cells present in mule seminiferous tubules. Although spermatogenesis in mules did not advance beyond secondary spermatocytes/newly formed round spermatids, germ cell associations in the seminiferous epithelium and pachytene spermatocytes nuclear volume in donkeys and mules were similar. The duration of spermatogenesis was estimated using intratesticular injections of tritiated thymidine. Each spermatogenic cycle in donkeys lasted 10.5 days. A similar value was found in mules ( approximately 10.1 days). Considering that the entire spermatogenic process takes approximately 4.5 cycles to be completed, its total duration in donkeys was estimated to last 47.2 days. The results found for mules suggest that the mechanisms involved in the determination of testis structure and function are probably originated from donkeys. Also, the data found for mules suggest that their seminiferous tubules are able to sustain complete spermatogenesis. In this regard, this species is a potential model for transplants of germ cells originated from donkeys and horses or other large animals.     

Chromosomal Assignment of Six Genes (EIF4G3, HSP90, RBBP6, IL8, TERT,and TERC) in Four Species of the Genus Equus

We mapped six genes (EIF4G3, HSP90, RBBP6, IL8, TERT, and TERC) on the chromosomes of Equus caballus, Equus asinus, Equus grevyi, and Equus burchelli by fluorescence in situ hybridization. Our results add six type I markers to the cytogenetic map of these species and provide new information on the comparative genomics of the genus Equus.     

Xenografting of adult mammalian testis tissue

Xenografting of testis tissue from immature males from several mammalian species to immunodeficient mouse hosts results in production of fertilization-competent sperm. However, the efficiency of testis tissue xenografting from adult donors has not been critically evaluated. Testis tissue xenografting from sexually mature animals could provide an option to preserve the genetic material from valuable males when semen for cryopreservation cannot be collected. To assess the potential use of this technique for adult individuals, testes from adult animals of six species (pig, goat, cattle, donkey, horse and rhesus monkey) were ectopically grafted to host mice. Grafts were recovered and analyzed at three time points: less than 12 weeks, between 12 and 24 weeks and more than 24 weeks after grafting. Histological analysis of the grafts revealed effects of species and donor tissue maturity: all grafts from species with greater daily sperm production (pig and goat) were found to have degenerated tubules or grafts were completely degenerated. None of the xenografts from mature adult bull and monkeys contained differentiated spermatogenic cells when examined more than 12 weeks post-grafting but tubules with Sertoli cells only remained. In grafts from a young adult bull, Sertoli cells persisted much longer than with the mature adult grafts. In grafts from a young adult horse, spermatogenesis proceeded to meiosis. In grafts from a young adult donkey and monkey, however, complete spermatogenesis was found in the grafts. These results show that testis tissue grafts from mature adult donors did not support germ cell differentiation but seminiferous tubules with Sertoli cells only survived in some species. The timing and progression of tubular degeneration after grafting of adult testis tissue appear to be related to the intensity of spermatogenesis at the time of grafting. Testis tissue from sub-adult donors survives better as xenograft than tissue from mature adult donors, and complete spermatogenesis can occur albeit with species-specific differences.