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Seroprevalences of antibodies to Neospora caninum and Toxoplasma gondii in zoo animals.

Neospora caninum is an apicomplexan parasite that causes neuromuscular disease in dogs and abortions in cattle. Little is known about the prevalence of antibodies to this parasite in zoo animals. Sera from 556 animals, from 13 Czech and Slovak zoos were tested for antibodies to N. caninum and Toxoplasma gondii by indirect fluorescent antibody test. Antibodies to N. caninum were found in 31 of 556 zoo animals (5.6%), representing 18 of 114 species tested: Eurasian wolf (Canis lupus lupus), Maned wolf (Chrysocyon brachyurus), fennec (Vulpes zerda), cheetah (Acinonyx jubatus), jaguarundi (Herpailurus yaguarondi), Eurasian lynx (Lynx lynx), Indian lion (Panthera leo goojratensis), fisher (Martes pennanti), blackbuck (Antilope cervicapra), European bison (Bison bonasus), lechwe (Kobus leche), African buffalo (Syncerus caffer caffer), eland (Taurotragus oryx), sitatunga (Tragelaphus spekei gratus), Thorold's deer (Cervus albirostris), Eastern elk (C. elaphus canadensis), Vietnam sika deer (C. nippon pseudaxis) and Père David's deer (Elaphurus davidianus). Titres ranged from 1:40 to 1:2560. The highest prevalence 50% was found in family mustelidae of the order carnivora. Antibodies to T. gondii were detected in 193 of 556 zoo animals (34.7%) representing 72 of 114 species tested, with titres ranging from 1:40 to 1:40960. The highest prevalence 100% was found in families: hyaenidae, mustelidae, ursidae and viveridae of the order carnivora. The results of this study indicate that zoo animals have more exposure to T. gondii than to N. caninum. It is the first report of seroprevalence of antibodies to N. caninum in European zoo animals.

Animals↗

Molecular phylogeny of the tribe Bovini (Bovidae, Bovinae) and the taxonomic status of the Kouprey, Bos sauveli Urbain 1937.

The kouprey is a very rare bovid species of the Indochinese peninsula, and no living specimen has been described for a long time, suggesting that it is possibly extinct. Its systematic position within the tribe Bovini remains confused since the analyses of morphological characters have led to several conflicting hypotheses. Some authors have also suggested that it could be a hybrid species produced by the crossing of the banteng with gaur, zebu, or water buffalo. Here we performed a molecular phylogeny of the tribe Bovini to determine the taxonomic status of the kouprey. DNA was extracted from the holotype specimen preserved in the MNHN collections. Phylogenetic analyses were carried out on a matrix including all the taxonomic diversity described in the tribe Bovini, and 2065 nucleotide characters, representing three different markers, i.e., the promotor of the lactoferrin and two mitochondrial genes (cytochrome b and subunit II of the cytochrome c oxidase). The results show that the kouprey belongs to the subtribe Bovina, and that three different clades can be evidenced into this group: the first includes the domestic ox, zebu, and European bison; the second incorporates the yak and American bison; and the third contains the kouprey, banteng and gaur. All hypotheses involving hybridization for the origin of the kouprey can be rejected, confirming that it is a real wild species. Molecular datings and biogeographic inferences suggest that the kouprey diverged from banteng and gaur during the Plio-Pleistocene of Asia. In addition, several molecular signatures were detected in the cytochrome b gene, permitting a molecular identification of the kouprey. We propose a conservation project based on a molecular taxonomy approach for tracking the kouprey in Indochina in order to determine whether some populations still survive in the wild.

Animals↗

Basic hematological values in some wild ruminants in captivity.

Analyses of hematological parameters were carried out on eight axis deer (Cervus axis), 12 fallow deer (Cervus dama), 16 red deer (Cervus elaphus hippelaphus), three sambar (Cervus unicolor), nine Père David deer (Elaphurus davidianus), 20 European bison (Bison bonasus), seven nilgai (Boselaphus tragocamelus), eight mouflon (Ovis musimon), four white-bearded gnu (Connochaetes taurinus) and six barbary sheep (Ammotragus lervia). The following parameters were determined: packed cell volume, hemoglobin concentration, red blood cell number, mean corpuscular volume, mean corpuscular hemoglobin, mean corpuscular hemoglobin concentration, leukocyte number and differential leukocyte count. Some degree of interspecific variability was found in the artiodactyls for these hematological parameters. An inverse relationship between mean corpuscular volume and red blood cell number is described, and specific values for these parameters are given for the four subfamilies.

Animals↗

Constitutive amplification of a zinc finger protein gene in cattle.

The human OZF gene encodes a protein consisting essentially of zinc finger motifs of the Krüppel type. Evolutionary studies revealed amplification of the bovine OZF gene in cattle. Domestic cattle includes two major subspecies: taurine (Bos primigenius taurus) and zebu (B. p. indicus). Amplified sequences were found in both subspecies and mapped to the same locus of chromosome X in band q11. No amplification was found in other bovids and in particular in Bison, the closest related genus to the genus Bos. One copy of the amplified locus was cloned and sequenced. It encodes a protein sharing 95% identity with the human OZF protein. The bovine OZF gene was detected in an additional locus on chromosome 18 in both Bos and Bison, in band q23-24, which is likely to represent the ancestral position of the OZF gene. This is the first evidence of gene amplification in a mammalian species during evolution.

Amino Acid Sequence↗

Microsatellite DNA variation and the evolution, domestication and phylogeography of taurine and zebu cattle (Bos taurus and Bos indicus).

Genetic variation at 20 microsatellite loci was surveyed to determine the evolutionary relationships and molecular biogeography of 20 different cattle populations from Africa, Europe and Asia. Phylogenetic reconstruction and multivariate analysis highlighted a marked distinction between humpless (taurine) and humped (zebu) cattle, providing strong support for a separate origin for domesticated zebu cattle. A molecular clock calculation using bison (Bison sp.) as an outgroup gave an estimated divergence time between the two subspecies of 610,000-850,000 years. Substantial differences in the distribution of alleles at 10 of these loci were observed between zebu and taurine cattle. These markers subsequently proved very useful for investigations of gene flow and admixture in African populations. When these data were considered in conjunction with previous mitochondrial and Y chromosomal studies, a distinctive male-mediated pattern of zebu genetic introgression was revealed. The introgression of zebu-specific alleles in African cattle afforded a high resolution perspective on the hybrid nature of African cattle populations and also suggested that certain West African populations of valuable disease-tolerant taurine cattle are under threat of genetic absorption by migrating zebu herds.

Africa, Western↗

Genetic characterization of the bovine leukaemia inhibitory factor (LIF) gene: isolation and sequencing, chromosome assignment and microsatellite analysis.

The bovine leukaemia inhibitory factor was isolated from a phage library and sequences for the gene, in addition to 1213 bp of 5' and 432 bp of 3' sequences, were obtained and compared with other mammalian leukaemia inhibitory factor genes. Comparisons indicated amino acid homologies ranging from 89.6% to 77.2% with the human and mouse homologues, respectively. Analysis of 500 bp of 5' regulatory regions indicated homologies ranging from 83.6% to 74.4% with the corresponding human and sheep sequences, respectively. Additionally, bovine leukaemia inhibitory factor-specific primers were prepared, and a panel of bovine x hamster somatic cell lines were analysed by the polymerase chain reaction (PCR). Data indicated 93% concordance of leukaemia inhibitory factor with aldehyde dehydrogenase 2 located on bovine chromosome 17, and concordance of 81% with myelin basic protein situated on bovine chromosome 24. Southern analysis of selected hybrids confirmed the PCR results, thus conclusively assigning the bovine leukaemia inhibitory factor gene to chromosome 17. Sequence analysis also revealed a microsatellite in intron 2 of the bovine leukaemia inhibitory factor. Analysis of this region by PCR in 22 unrelated Bos taurus and 19 unrelated Bos indicus cattle detected nine different alleles. Polymorphic information content values were 0.53 and 0.80 in B. taurus and B. indicus, respectively. Additionally, the same leukaemia inhibitory factor primers successfully detected allelic variants at this locus in Bos javanicus, Bos guarus and Bison bison but not in Odocoileus virginianus.

Amino Acid Sequence↗

Close genetic relationship of early neolithic cattle from Ziegelberg (Freising, Germany) with modern breeds.

In 2003 a variety of crafts and bone specimens were found during excavations of a Neolithic settlement near Freising, the southernmost site of the Linear Pottery Culture in Bavaria. Six cattle bones were used to extract ancient DNA (aDNA). Applying nested and touchdown PCR, two fragments of the mitochondrial (mt) DNA control region could be amplified from specimen 533/III which yielded a total of 230 base pairs (bp). The sequence was compared with the homologous part of 40 extant breeds of Bos taurus and B. indicus and related species, such as Banteng (B. javanicus), Gaur (B. gaurus), the European bison (Bison bonasus) and the aurochs (B. primigenius). A neighbour joining tree was constructed based on the appropriate model of sequence evolution. The control region sequence of the 533/III cattle bone, whose age was determined by radiocarbon dating, clusters close to the extant European breeds, but distinctly apart from the basal aurochs and far distant from the B. indicus group. The archaeological and genetic analyses of Bos Ziegelberg demonstrate that domesticated cattle reached southern Bavaria at least 7000 years ago.

Animals↗

Monomorphism and polymorphism at Mhc DRB loci in domestic and wild ruminants.

Genetic polymorphism at Mhc class II DRB loci was investigated in samples of musk-ox from Canada and Greenland; moose from Sweden, Norway, Canada, and Alaska; roe deer from Norway and Sweden; reindeer from Svalbard and Norway; fallow deer from Norway and Sweden; and red deer from Norway. The results were compared with published data on cattle, bison, goat, sheep, and red deer. Cattle-specific primers amplified a single DRB locus in all species except fallow deer and red deer, in which two loci were found. Single strand conformation polymorphism analysis and DNA sequence analysis were employed to detect genetic polymorphism. Complete monomorphism was found in musk-ox and fallow deer. Limited polymorphism was found in the moose, roe deer, and reindeer from Svalbard, whereas intermediate to extensive DRB diversity was present in reindeer from Norway and in bison, sheep, goat, cattle, and red deer. The restricted Mhc diversity in moose, roe deer, and fallow deer is notable in relation to the dramatic population expansion of moose and roe deer in Sweden during this century and since fallow deer is used for meat and game production with good results and without any marked disease problems. The results question the view that species or populations with restricted Mhc diversity have poor resistance to infectious diseases. A phylogenetic tree analysis revealed a clustering of DRB sequences within species rather than within allelic lineages across species. The results suggest trans-species persistence of polymorphic sequence motifs rather than of allelic lineages.

Amino Acid Sequence↗

Development, testing and commercialization of a new brucellosis vaccine for cattle.

Vaccines used against brucellosis do not generally protect completely against infection or abortion. Genetic analysis has revealed differences in arrangements of DNA sequences between these vaccine strains and the virulent parent strain and permits the specific identification of field isolates of B. abortus as wild-type or vaccine strain. B. abortus strain 19 is a low-virulence, live vaccine developed for use in cattle. Although it is effective, strain 19 vaccine had a tropism for the placenta and caused abortion when given to pregnant cows, was infectious for humans, and caused serologic responses in calves that could not be differentiated from those in cattle infected with natural field strains. In the mid-1980s the need for a new vaccine emerged when the USDA increased its efforts in brucellosis eradication. In the 1990s, research on biosafety, vaccine efficacy and field application rapidly established the fact that strain RB51 is protective in cattle at doses comparable to those of strain 19. Thus, Brucella abortus strain RB51 is the vaccine of choice against brucellosis of cattle in the United States. Studies have established the relative efficacy of strain RB51 vaccine on bison, and the vaccine has also been accepted for use in commercial bison herds in the U.S.

Abortion, Veterinary↗

Occurrence and diurnal population fluctuations of the ruminal protozoan Microcetus lappus.

A series of experiments with bison and cattle were conducted to obtain information on the relatively uncommon ruminal protozoan Microcetus lappus. Although M. lappus is a holotrich, diurnal changes in concentrations indicate that it follows a cycle unlike most other holotrichs, decreasing shortly after feed is offered and then gradually increasing over time. Concentrations of M. lappus varied widely among animals, exceeding 50% of the total protozoan population in some cattle. In bison, Microcetus concentrations averaged 2% of the protozoan population. Dietary protein and energy levels apparently did not influence Microcetus numbers. The highest concentrations of M. lappus were found in the reticulum, whereas the lowest numbers occurred in the mid-dorsal sac.

Animals↗

Comparative transcriptional analysis of human macrophages exposed to animal and human isolates of Mycobacterium avium subspecies paratuberculosis with diverse genotypes.

Mycobacterium avium subsp. paratuberculosis is the causative agent of Johne's disease in animals and has been hypothesized to be associated with Crohn's disease in humans. Recently, M. avium subsp. paratuberculosis isolates recovered from Crohn's disease patients were shown to have limited diversity, implying the existence of human disease-associated genotypes and strain sharing with animals (A. H. Ghadiali et al., J. Clin. Microbiol. 42:5345-5348, 2004). To explore whether these genotypic differences or similarities among human and animal isolates translated to functionally significant attributes such as variance in host preference and/or difference in magnitude of infections, we performed a global scale analysis of M. avium subsp. paratuberculosis isolates that were representative of different genotypes and host species using DNA microarrays. Genome-wide characterization of the transcriptional changes was carried out using a human monocytic cell line (THP-1 cells) in response to different genotypes of M. avium subsp. paratuberculosis isolates recovered from various hosts. We identified several differentially expressed genes during early intracellular infection, including those involved in common canonical pathways such as NF-kappaB, interleukin-6 (IL-6), mitogen-activated protein kinase/extracellular signal-regulated kinase, and Jun N-terminal protein kinase signaling, as well as genes involved in T helper type 1 (Th1) responses (such as CCL5 ligand) and those that encode several proinflammatory cytokines and chemokine receptors. The cattle and human isolates of M. avium subsp. paratuberculosis, regardless of their short sequence repeat (SSR) genotype, induced similar global gene expression patterns in THP-1 cells. They differentially regulated genes necessary for cell survival without causing major alterations in proinflammatory genes. In contrast, the sheep isolates representing diverse SSR genotypes closely resembled the global gene expression pattern of an M. avium subsp. avium isolate, and they significantly up-regulated proinflammatory genes related to IL-6, T-cell receptor, B-cell receptor, and death receptor signaling within THP-1 cells. Additionally, we demonstrated consistency among infecting genotypes of M. avium subsp. paratuberculosis isolated from diverse hosts [cattle (n=2), human (n=3), sheep (n=2), and bison (n=1)] in quantitative reverse transcription-PCR analysis of seven differentially expressed genes. While the levels of expression induced by the bison isolate were different compared with cattle or human isolates, they followed the common anti-inflammatory, antiapoptotic trend. Our data suggest that the macrophage responses to M. avium subsp. paratuberculosis isolates from cattle and human sources, regardless of genotype, follow a common theme of anti-inflammatory responses, an attribute likely associated with successful infection and persistence. However, these expression patterns differ significantly from those in THP-1 cells infected with sheep isolates of M. avium subsp. paratuberculosis or the M. avium subsp. avium isolate. These data provide a transcriptional basis for a variety of pathophysiological changes observed during early stages of infection by different strains of M. avium subsp. paratuberculosis, a first step in understanding trait-allele association in this economically important disease.

Animals↗

Isolation from Pasteurella multocida of a lipopolysaccharide antigen with immunizing and toxic properties.

A heat-stable, particulate, lipopolysaccharide-protein antigenic complex has been isolated from a virulent, encapsulated strain of Pasteurella multocida by extraction with cold, formalinized saline, and centrifugation at 105,000 x g. The original bacterial culture had been obtained from a bison that died of hemorrhagic septicemia, an infectious disease of cattle and buffalo. Injection of fractional milligram amounts of the antigen into mice, rabbits, and calves produced toxic reactions which frequently resulted in death of the host. The surviving animals demonstrated a high degree of immunity to challenge with live, virulent organisms. Two injections with 15 mug of the antigen produced a high degree of immunity in mice without the development of any signs of toxicity. The gross chemical composition and toxicity of the antigen were similar to those reported for endotoxins obtained by the Boivin or Westphal procedure. Although strong serological cross-reactions were obtained in Ouchterlony plates between the 105,000 x g antigens from the bison strain and an avian strain with antisera to these strains, these antisera agglutinated only the bacterial cells of the homologous strain. The active immunity obtained in mice by the injection of the 105,000 x g antigens of each strain was specific and could be correlated with the agglutination tests.

Animals↗

Sterility in hybrid cattle. I. Distribution of constitutive heterochromatin and nucleolus organizer regions in somatic and meiotic chromosomes.

The distribution of constitutive heterochromatin and nucleolus organizer regions (NOR's) in somatic as well as in meiotic chromosomes of Bos taurus, Bos banteng, Bison bison, and their hybrids are analyzed. C-bands are present in the centromeric regions of every autosome. The X chromosome does not show a distinct C-band in the centromeric region, whereas the Y chromosome contains an appreciable amount of C-band material. In somatic metaphases, NOR's are present on the telomeric ends of five pairs of autosomes. During pachytene, five autosomal bivalents contain NOR's on their terminal ends. Meiotic preparations made from sterile bulls did not contain stages beyond the degenerating pachytene, which are C-banding, more frequently showed clustering of heterochromatin than did the pachytene stage in normal bulls.

Animals↗

[Specific molecular markers of the rust resistance gene M4 in flax].

Flax (Linum usitatissimum L.) is an important fiber and oil-producing crop. Flax rust, caused by Melampsora lini Ehrenb. Lev., occurs worldwide and can cause severe losses in seed yield and fiber quality. In order to identify molecular markers linked to the flax rust resistant gene M4, RAPD analysis of NM4, a near-isogenic line containing the M4 gene, and the recurrent parent Bison was carried out with 540 decamer primers. The primer OPA18 could stably amplify a specific fragment, OPA18(432), in the NM4 line. The OPA18(432) marker was testified to be closely linked to the M4 gene with a genetic distance of 2.1 cM through the analysis of the F2 mapping population derived from a cross of Bison x NM4. Based on the sequence of OPA18(432), the specific PCR primers were designed, and a SCAR marker for the M4 gene was produced. Amplification of different resistant materials proved that the maker is specific for the M4 gene. This marker has been used successfully in marker-assisted selection in the flax breeding program.

Base Sequence↗

MilkProtChip--a microarray of SNPs in candidate genes associated with milk protein biosynthesis--development and validation.

MilkProtChip is an oligonucleotide microarray based on the arrayed primer extension (APEX) technique, allowing genotyping of single nucleotide polymorphisms (SNPs) in genes of interest for bovine milk protein biosynthesis. APEX consists of a sequencing reaction primed by an oligonucleotide anchored with its 5'end to a glass slide and terminating one nucleotide before the polymorphic site. The extension with one fluorescently labeled dideoxy nucleotide complementary to the template reveals the polymorphism. A total of 75 SNPs were selected among those associated directly or potentially with milk protein content. Among the 75 SNPs, 4 did not produce a positive signal. Most of the remaining SNPs produced a signal for both strands, except for 4 (one strand). In the validation step, 12 Polish Holstein bulls, 1 Polish Red bull, 1 bison (Bison bonasus), 11 Jersey cows and 25 Polish Holstein cows were screened to validate SNPs. Among the 71 selected SNPs--26 were found monoallelic, the rest showing at least two genotypes for the entire population under study. All the animals were earlier genotyped for 2-5 SNPs by PCR-RFLP and PCR sequencing and all showed complete concordance with APEX genotyping. APEX reactions showed relatively high signal frequencies: more than 0.9, 0.9-0.8 and below 0.8, for 65, 4 and 2 DNA samples, respectively. The primary application of the MilkProtChip is the simultaneous genotyping of dozens of SNPs to reveal and clarify the genetic background of milk protein biosynthesis. The chip may possibly be used for dairy cattle identification and paternity analysis, evolutionary studies, the evaluation of genetic distances between wild and domestic cattle breeds and the domestication history of bovine species.

Animals↗

[Expression of genes controlling modification of amylase in hybrids and original representatives of a cattle subfamily (Bovinae)].

Modifications of similar type were noted in the number of multiple AMY-1 forms of amylase isozyme for various species of Artiodactyla. It is supposed that these modifications are linked to periodical activation during evolution of genes responsible for modification of the molecules. Data obtained on Bovinae hybrids testify to this point of view. Inactivation of active and reinactivation of "silent" genes responsible for modification of AMY-1 molecules are observed in hybrids of bison x cow and bison x aurochs combinations.

Amylases↗

Fluoride toxicosis in wild ungulates.

To compare the occurrence of chronic fluoride toxicosis in wild and domestic animals in selected areas of Utah, Idaho, Montana, and Wyoming, deer, elk, and bison bones and teeth were collected for evaluation. Vegetation and drinking water samples also were collected, so that potential sources of fluoride could be evaluated. Deer, elk, and bison were found to be susceptible to the adverse effects of ingestion of excessive amounts of fluoride. Teeth and bones were primarily affected with characteristic lesions. Pathognomonic soft tissue changes were not observed. The animals had been exposed to a variety of sources of excessive fluoride, including water high in fluoride, forages contaminated by industrial effluents that were high in fluoride, vegetation contaminated with high fluoride-content soil by rain splash or wind, or a combination of these sources. Waters high in fluoride, especially from geothermal springs and wells, often contained appreciable amounts of various soluble salts. Evidence accumulated from specimens collected throughout the aforementioned states indicated that there are areas where chronic fluoride toxicosis is a problem for wildlife. These areas were where natural sources of fluorine (especially geothermal waters) provided amounts for ingestion that exceed species tolerance limits or were near certain industrial operations.

Animals↗

Hyperdisease in the late Pleistocene: validation of an early 20th century hypothesis.

The hypothesis of disease-related large mammal extinction has new support. A unique pathologic zone of resorption was first noticed in a Hiscock Mammut americanum metacarpal. The pathognomonic zone of resorption was present in fifty-nine (52%) of 113 skeletons with feet available for examination. Metacarpals and metatarsals were most commonly affected. Associated rib periosteal reaction is highly suggestive of tuberculosis and the foot lesions were identical to that documented in Bison as pathognomonic for tuberculosis. Recognizing that only a portion of animals infected by infectious tuberculosis develop bone involvement, the high frequency of the pathology in M. americanum suggests that tuberculosis was not simply endemic, but actually pandemic, a hyperdisease. Pandemic tuberculosis was one of several probable factors contributing to mastodon extinction.

Animal Diseases↗