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Is the decline of desert bighorn sheep from infectious disease the result of low MHC variation?

Bighorn sheep populations have greatly declined in numbers and distribution since European settlement, primarily because of high susceptibility to infectious diseases transmitted to them from domestic livestock. It has been suggested that low variation at major histocompatibility complex (MHC) genes, the most important genetic aspect of the vertebrate immune system, may result in high susceptibility to infectious disease. Therefore, we examined genetic polymorphism at a MHC gene (Ovca-DRB) in a large sample, both numerically and geographically, of bighorn sheep. Strikingly, there were 21 different alleles that showed extensive nucleotide and amino acid sequence divergence. In other words, low MHC variation does not appear to be the basis of the high disease susceptibility and decline in bighorn sheep. On the other hand, analysis of the pattern of the MHC polymorphism suggested that nonsynonymous substitutions predominated, especially at amino acids in the antigen-binding site. The average overall heterozygosity for the 16 amino acid positions that are part of the antigen binding site is 0.389 whereas that for the 67 amino acid positions not involved with antigen binding is 0.076. These findings imply that the diversity present in this gene is functionally significant and is, or has been, maintained by balancing selection. To examine the evolution of DRB alleles in related species, a phylogenetic analysis including other published ruminant (Bovidae and Cervidae) species, was carried out. An intermixture of sequences from bighorn sheep, domestic sheep, goats, cattle, bison, and musk ox was observed supporting trans-species polymorphism for these species. To reconcile the species and gene trees for the 104 sequences examined, 95 'deep coalescent' events were necessary, illustrating the importance of balancing selection maintaining variation over speciation events.

Amino Acid Sequence↗

Mitochondrial diversity and the origins of African and European cattle.

The nature of domestic cattle origins in Africa are unclear as archaeological data are relatively sparse. The earliest domesticates were humpless, or Bos taurus, in morphology and may have shared a common origin with the ancestors of European cattle in the Near East. Alternatively, local strains of the wild ox, the aurochs, may have been adopted by peoples in either continent either before or after cultural influence from the Levant. This study examines mitochondrial DNA displacement loop sequence variation in 90 extant bovines drawn from Africa, Europe, and India. Phylogeny estimation and analysis of molecular variance verify that sequences cluster significantly into continental groups. The Indian Bos indicus samples are most markedly distinct from the others, which is indicative of a B. taurus nature for both European and African ancestors. When a calibration of sequence divergence is performed using comparisons with bison sequences and an estimate of 1 Myr since the Bison/Bos Leptobos common ancestor, estimates of 117-275,000 B.P. and 22-26,000 B.P. are obtained for the separation between Indians and others and between African and European ancestors, respectively. As cattle domestication is thought to have occurred approximately 10,000 B.P., these estimates suggest the domestication of genetically discrete aurochsen strains as the origins of each continental population. Additionally, patterns of variation that are indicative of population expansions (probably associated with the domestication process) are discernible in Africa and Europe. Notably, the genetic signatures of these expansions are clearly younger than the corresponding signature of African/European divergence.

Africa↗

Fluorescence polarization assay for the diagnosis of brucellosis: a review.

Fluorescence polarization assay (FPA) is based on the rotational differences between a small soluble antigen molecule in solution (labelled with a fluorochrome) and the antigen molecule complexed with its antibody. A small molecule will rotate randomly at a rapid rate, resulting in rapid depolarization of light, while a larger complex molecule will rotate slower and depolarize light at a reduced rate. The rate change in depolarization can be measured. The FPA is a homogeneous assay which does not require removal of unreacted reagents and can, therefore, be performed very quickly and, given portable equipment, in the laboratory and in the field. The latter obviates the need for shipping samples and eliminates waiting for results, as well as reducing test costs. The FPA technology has been developed and validated for the serological diagnosis of brucellosis in cattle, swine, sheep, goats, bison, and cervids. Sufficient cross reactivity of the common epitopes of Brucella abortus, B. melitensis and B. suis O-polysaccharide (OPS) allowed for the use of a single antigen for all species of smooth Brucella and animals. The OPS prepared from B. abortus S1119.3 was conjugated with fluorscein isothiocyanate (FITC). The FPA was initially developed for testing serum; however, the technology has been extended to testing whole blood and milk from individual animals or bulk tank samples pooled from 2000 or fewer animals. The accuracy of the FPA equalled or exceeded those obtained using other serological tests such as the buffered antigen plate agglutination test (BPAT), the milk ring test (MRT), the complement fixation test (CFT), the indirect enzyme immunoassay (IELISA), and the competitive enzyme immunoassay (CELISA).

Animals↗

Are there preconceptional determinants of mammalian sex? A response to Boklage (2005).

Boklage (2005, Hum Reprod 20,583-587) wrote: 'Changes in, or mediated by, the epigenetic environment of embryogenesis provide the most plausible prospects for causes of changes in secondary sex ratio'. Without impugning this notion, I suggest here that other causes of variation in secondary sex ratio antedate fertilization or, in other words, that there are circumstances under which unequal numbers of male and female zygotes are formed. It will be documented here that this suggestion has repeatedly been made on the basis of data on many mammalian species (golden hamster, sheep, mouse, white-tailed deer, American bison, springbok, domestic cattle and Barbary macaque). It may be acknowledged that the causes of this hypothesized preconceptional variation are not established. I have suggested one (parental hormones around the time of conception). But the truth of that hypothesis is not germane to the thrust of the present argument--which is to document the strong suspicions that such preconceptional determinants do, indeed, exist. Efforts should be made to confirm or discredit these suspicions.

Animals↗

Drawing pedigrees.

The problem of drawing a pedigree in a clear and concise manner is discussed and a case is made for the adoption of the marriage node graph as the most appropriate format in which to depict highly looped pedigrees. A method for assigning suitable coordinates to marriages and individuals is then described. This involves use of simulated annealing to search for the clearest picture. The method is illustrated by two examples: the pedigree of Przewalski's Horse and that of the European Bison.

Animals↗

Cytochrome b gene haplotypes characterize chromosomal lineages of anoa, the Sulawesi dwarf buffalo (Bovidae: Bubalus sp.).

Partial mitochondrial cytochrome b gene sequences reveal two deeply differentiated mtDNA lineages in anoa dwarf buffaloes (Bubalus depressicornis) from the studbook herd in European zoos. Three matrilinear lineages of lowland anoas (depressicornis type) contributed three rather similar sequence haplotypes, but one remarkably distinct haplotype was observed exclusively in mountain anoas (quarlesi type) descended from one founder female. The carriers of the distinctive mtDNA haplotype were also distinguished by several chromosomal and phenotypic peculiarities too. The differentiation between the mtDNA lineages of anoa approached or even surpassed the genetic divergence between some uncontested species of wild cattle. The depth of this haplotype divergence in anoas is discussed against the background of the phylogenetic age of these paleoendemic inhabitants of a predator-free island refugium, Sulawesi, who are among the most plesiomorphic living bovines. The studbook breeding of captive anoas as a safeguard against extinction might profit from such population genetic markers. These cytochrome b gene sequences were unable to resolve the phylogeny of nine bovine taxa robustly, except the divergence of Bubalus, Synceros, Bison, and Bos (sensu lato) genera.

Animals↗

African-derived mitochondria in South American native cattle breeds (Bos taurus): evidence of a new taurine mitochondrial lineage.

This article reports the nucleotide diversity within the control region of 42 mitochondrial chromosomes belonging to five South American native cattle breeds (Bos taurus). Analysis of these data in conjunction with B. taurus and B. indicus sequences from Africa, Europe, the Near East, India, and Japan allowed the recognition of eight new mitochondrial haplotypes and their relative positions in a phylogenetic network. The structure of genetic variation among different hypothetical groupings was tested through the molecular variance decomposition, which was best explained by haplotype group components. Haplotypes surveyed were classified as European-related and African-related. Unexpectedly, two haplotypes within the African cluster were more divergent from the African consensus than the latter from the European consensus. A neighbor-joining tree shows the position of two haplotypes compared to European/African mitochondrial lineage splitting. This different and putatively ancestral mitochondrial lineage (AA) is supported by the calibration of sequence divergence based on the Bos-Bison separation. The European/African mitochondria divergence might be subsequent (67,100 years before present) to that between AA and Africans (84,700 years before present), also preceding domestication times. These genetic data could reflect the haplotype distribution of Iberian cattle five centuries ago.

Africa↗

Genomic structure and tissue-specific expression of human and mouse genes encoding homologues of the major bovine seminal plasma proteins.

Sperm capacitation is a maturation event that takes place in the female reproductive tract and is essential for fertilization. A family of phospholipid-binding proteins present in bovine seminal plasma (BSP proteins) binds the sperm membrane at ejaculation and promotes bovine sperm capacitation. Homologues of these proteins have also been isolated from boar, ram, goat, bison and stallion seminal fluid, suggesting that BSP proteins and their homologues are conserved among mammals. However, there have been no reports on BSP-homologous proteins in mice and humans to date. A search of the mouse and human genomes, using the nucleic acid sequences of BSP proteins, revealed the presence of three BSP-like sequences in the mouse genome, named mouse BSP Homologue 1 (mBSPH1), mBSPH2 and mBSPH3, and one sequence in the human genome (hBSPH1). Mouse epididymal expressed sequence tags corresponding to partial sequences of mBSPH1 and mBSPH2 were identified. The entire complementary DNA (cDNA) sequences of mBSPH1 and mBSPH2 from mouse epididymis and hBSPH1 from human epididymis were obtained by 5'-/3'-rapid amplification of cDNA ends (RACE) and encode predicted proteins containing two tandemly repeated fibronectin type II domains, which is the signature of the BSP family of proteins. Using RT-PCR, it was revealed that mBSPH1, mBSPH2 and hBSPH1 mRNA are expressed only in the epididymis. Expression of mBSPH3 was not detected in any tissue and probably represents a pseudogene. This work shows, for the first time, that BSP homologues are expressed in mouse and human and may be involved in sperm capacitation in these species.

Adult↗

Evolutionary affinities of the enigmatic saola (Pseudoryx nghetinhensis) in the context of the molecular phylogeny of Bovidae.

To elucidate the systematic status of the enigmatic saola (Pseudoryx nghetinhensis), a new bovid genus recently discovered in Vietnam, and to investigate phylogenetic relationships within the family Bovidae, four distinct DNA markers were sequenced. Complete mitochondrial cytochrome b (1143 bp) and 12S rRNA (956 bp) genes and non-coding regions from the nuclear genes for aromatase cytochrome P-450 (199 bp) and lactoferrin (338 bp) have been compared for 25 bovid species and three Cervidae and Antilocapridae outgroups. Independent and/or combined analyses of the four nucleotide matrices through maximum parsimony and maximum-likelihood methods indicated that Bovidae consists of two major lineages, i.e. Bovinac which contains the tribes Bovini, Boselaphini and Tragelaphini, and Antilopinae which encompasses all other bovids. Within Bovinae, the tribe Bovini is divided into buffalo Bovini (Bubalus and Syncerus) and cattle Bovini (Bos and Bison) and Tragelaphini are possibly related to Boselaphini. Pseudoryx is shown to be (i) robustly nested within Bovinae; (ii) strongly associated with Bovini; and (iii) tentatively sharing a sister-group relationship with cattle Bovini. Within Antilopinae, three robust clades are in evidence: (i) Hippotragus and Damaliscus are linked to Ovis; (ii) Aepyceros joins Neotragus; and (iii) Cephalophus clusters with Oreotragus.

Animals↗

Immunological reactions from fossil material.

Genetic relations among living species can be deduced from biochemical as well as morphological similarities, but our understanding of fossil species has depended entirely on their morphology. Residual proteins in fossils might provide genetic information, but their small quantity and chemical alterations due to time and environmental agents have prevented the obtaining of species-specific analysis. This report describes a radioimmunoassay capable of detecting extremely small amounts of fossil proteins, such as collagen and albumin. Species-specific proteins have been identified in a frozen Siberian mammoth, a Pleistocene bison, and a series of human fossils that includes Neanderthal, Homo erectus and Australopithecus robustus. This technique promises to provide molecular data on the genetic affinities of fossil and living species.

Albumins↗

Experimental infection of sheep with ovine herpesvirus 2 via aerosolization of nasal secretions.

Ovine herpesvirus 2 (OvHV-2) is the causative agent of sheep-associated malignant catarrhal fever in clinically susceptible ruminants, including cattle, bison and deer. Studies of OvHV-2 have been hampered by the lack of an in vitro propagation system. Here, the use of nasal secretions collected from OvHV-2-infected sheep experiencing intense virus shedding episodes as a source of infectious virus for experimental animal infections was examined. OvHV-2 uninfected sheep were nebulized with nasal secretions containing approximately 10(8) to 10(1) copies of OvHV-2 DNA. The time to detectable viral DNA in peripheral blood leukocytes (7-12 days post-infection) and virus-specific antibody in plasma (9-32 days post-infection) varied with the dose of inocula administered. Here, the use of nasal secretions as a source of infectious OvHV-2 was defined and the minimum infectious dose of a pool of nasal secretions that can be used in further studies of viral pathogenesis and vaccine development was determined.

Aerosols↗

A novel subgroup of rhadinoviruses in ruminants.

In the course of investigating the malignant catarrhal fever (MCF) subgroup of rhadinoviruses, seven novel rhadinoviruses were identified in a variety of ruminants, including domestic sheep, bighorn sheep, bison, black-tailed deer, mule deer, fallow deer, elk and addax. Based on the DNA polymerase gene sequences, these newly recognized viruses clustered into a second distinct subgroup in ruminants with three members identified previously in cattle, goats and oryx. Phylogenetic analysis revealed that the currently known ruminant rhadinoviruses appear to comprise three distinct genetic lineages: (i) the MCF subgroup, defined by sequence identity and the presence of the 15A antigenic epitope; (ii) a second distinct subgroup, devoid of the 15A epitope, which contains the previously reported bovine lymphotropic herpesvirus and related viruses; and (iii) a third distinct subgroup represented by Bovine herpesvirus 4. Comparison of phylogenetic trees between the rhadinoviruses and their corresponding hosts further supports the gammaherpesvirus and host co-evolution theory.

Animals↗

Complete sequence and analysis of the ovine herpesvirus 2 genome.

Ovine herpesvirus 2 (OvHV-2) is endemic in sheep populations worldwide and causes malignant catarrhal fever (MCF), a lymphoproliferative disease, in cattle, bison and deer. OvHV-2 has been placed in the gammaherpesvirus subfamily and is related closely to Alcelaphine herpesvirus 1 (AlHV-1). Here, the cloning, sequencing and analysis of the complete OvHV-2 genome derived from a lymphoblastoid cell line from an affected cow (BJ1035) are reported. The unique portion of the genome consists of 130,930 bp, with a mean G+C content of 52 mol%. The unique DNA is flanked by multiple copies of terminal repeat elements 4205 bp in length, with a mean G+C content of 72 mol%. Analysis revealed 73 open reading frames (ORFs), the majority (62) of which showed homology to other gammaherpesvirus genes. A further subset of nine ORFs is shared with only the related AlHV-1. Three ORFs are entirely unique to OvHV-2, including a spliced homologue of cellular interleukin-10 that retains the exon structure of the cellular gene. The sequence of OvHV-2 is a critical first step in the study of the pathogenesis and treatment of MCF.

Animals↗

Changes in the Ribonuclease Activity of Flax Cotyledons following Inoculation with Flax Rust.

There was a significant increase in the ribonuclease activity of both resistant (Bombay) and susceptible (Bison) varieties of flax (Linum usitatissimum L.) 3 to 4 days after inoculation with flax rust (Melampsora lini [Pers.] Lev., race No. 3). A second and much greater increase in the activity of this enzyme occurred only in the susceptible host at later stages of disease development. While a similar increase in ribonuclease level was also caused by mechanical injury, evidence is presented showing qualitative differences between the enzyme from parasitized tissue and that from the mechanically injured cotyledons. Comparison of the enzyme from healthy and inoculated cotyledons and from flax rust revealed the presence of a relatively unstable component and some unique catalytic properties in the enzyme from inoculated cotyledons.

Journal Article↗

Changes in two ribonuclease isozymes during rust infection of flax cotyledons.

A ribonuclease fraction previously purified from flax by gel filtration was further resolved into two components by hydroxyl apatite chromatography. These were homogeneous with respect to electrophoresis and isoelectric focusing. Both enzymes are of RNase I type but differ in substrate specificity, kinetic properties, pH response, and isoelectric point.The two RNase isozymes show consistent properties when extracted from variety Bison (susceptible) or variety Bombay (resistant) with or without infection with race 3 of flax rust. The relative amounts of these isozymes change markedly during infection. These observations provide an explanation for the apparent qualitative changes in RNase noted previously. Differences between susceptible and resistant reactions in the early stages of disease are discussed.

Journal Article↗

Malignant catarrhal fever virus. Characterization of a United States isolate and development of diagnostic assays.

Malignant catarrhal fever (MCF) is a severe lymphoproliferative disease of certain domestic and wild ruminants. Two distinct but closely related viruses cause clinically indistinguishable syndromes in susceptible ruminant species: wildebeest-associated MCF virus (WA-MCFV) and sheep-associated MCF virus (SA-MCFV). Neither the pathogenesis nor the epidemiology of SA-MCF is understood, primarily because of a lack of adequate detection methods for the etiologic agent or antibody against that agent. Work designed to develop these tests has been under way in our laboratory. To obtain basic information about the virus, the in vitro growth properties of a US isolate of MCF virus were studied and its major viral proteins identified and characterized by a panel of monoclonal antibodies generated against the isolate. A monoclonal antibody to a broadly conserved epitope of MCF virus was identified, and a competitive-inhibition ELISA (CI-ELISA) was developed for detection of anti-MCF antibody in sheep and other ruminants. The monoclonal antibody (15-A) reacted with an epitope located on a glycoprotein complex, which was present in all isolates of MCF virus examined. Antibody from a wide variety of ruminants infected with MCF virus of both sheep and wildebeest origin competed with the monoclonal antibody 15-A for the epitope, which was not present on 14 other common ruminant viruses. The assay detected antibody in inapparently infected sheep, and in cattle, deer, and bison with clinical MCF. A PCR assay for DNA of the sheep-associated virus was developed, based on previously reported primers. Comparative studies demonstrated that the CI-ELISA was specific for MCFV antibody and that the PCR was more reliable for diagnosis of clinical MCF.

Animals↗

Diverse plant and animal genetic records from Holocene and Pleistocene sediments.

Genetic analyses of permafrost and temperate sediments reveal that plant and animal DNA may be preserved for long periods, even in the absence of obvious macrofossils. In Siberia, five permafrost cores ranging from 400,000 to 10,000 years old contained at least 19 different plant taxa, including the oldest authenticated ancient DNA sequences known, and megafaunal sequences including mammoth, bison, and horse. The genetic data record a number of dramatic changes in the taxonomic diversity and composition of Beringian vegetation and fauna. Temperate cave sediments in New Zealand also yielded DNA sequences of extinct biota, including two species of ratite moa, and 29 plant taxa characteristic of the prehuman environment. Therefore, many sedimentary deposits may contain unique, and widespread, genetic records of paleoenvironments.

Animals↗

Modulation of diversity by grazing and mowing in native tallgrass prairie

Species diversity has declined in ecosystems worldwide as a result of habitat fragmentation, eutrophication, and land-use change. If such decline is to be halted ecological mechanisms that restore or maintain biodiversity are needed. Two long-term field experiments were performed in native grassland to assess the effects of fire, nitrogen addition, and grazing or mowing on plant species diversity. In one experiment, richness declined on burned and fertilized treatments, whereas mowing maintained diversity under these conditions. In the second experiment, loss of species diversity due to frequent burning was reversed by bison, a keystone herbivore in North American grasslands. Thus, mowing or the reestablishment of grazing in anthropogenically stressed grasslands enhanced biodiversity.

Journal Article↗