Toxoplasmosis in nilgais (Boselaphus tragocamelus) and a saiga antelope (Saiga tatarica).

Three captive female nilgais (Boselaphus tragocamelus) either showed late-term abortion or their newborn calves died within 2 days of birth. Toxoplasma gondii DNA was demonstrated in the brain and liver of each fetus and in one of the two neonates by single-stage polymerase chain reaction (PCR) with TGR1E and by seminested PCR with B1 gene. Retrospectively, antibodies titers > or =640 to T. gondii by indirect fluorescence were found in the sera of all three female and one male nilgais. No other cause of abortion was detected. Fatal toxoplasmosis was also diagnosed in one captive, adult female saiga antelope (Saiga tatarica), which died suddenly. Toxoplasma gondii was detected in the liver, lung, spleen, kidney, and intestine. An unusual finding was the presence of numerous tissue cysts in the liver of this animal. Toxoplasmosis was confirmed by PCR with TGR1E and immunohistochemically. Toxoplasmic hepatitis and pneumonia were considered to be the primary causes of death.     

Osteomorphological features of the hind limb bones of Saiga antelope (Saiga tatarica)

The intralimb indices and calcaneal linear metrics are known as the reliable predictors of locomotor adaptation in artiodactyls. The osteological features of hindlimb in adult Saiga (Saiga tatarica) were described, and its correlation with cursoriality and habitat adaptation was discussed. Gross anatomy data showed Saiga owned the deep acetabulum as a broad lunate surface, the large acetabular anteversion, the well-developed ischiatic tuberosity and the prominent gluteal lines. It also presented the robust rough line and the strong gluteal tuberosity. A proximodistally elongated eminence located on the cranially distal tibia, which had not been found in goat. The tibial extensor groove was deep. The calcaneal tuberosity was robust. Digital anatomy data showed Saiga owned the higher metatarsal-femur ratio than forest musk deer and sheep. Comparing with wild bovids and sheep, Saiga presented a transitional variation in calcaneal form. The mean greatest length of the calcaneus (GLC) and the height of the sustentacular facet (HSF) in Saiga were shorter than that in sheep and longer than that in wild bovids respectively (F = 587.492; F = 10.264, p < .05). The wild bovids had longer cubonavicular facets than the other two groups (F = 18.587, p < .05). The great metatarsal–femur ratio of Saiga implied a superior cursorial ability and high conservation confronting the different habitats. The calcaneal linear metrics might shed light on lifestyle-related functional adaptation over decades of short-term evolution in the semi-free range environment.     

Application of bovine microsatellite markers for genetic diversity analysis of European bison (Bison bonasus)

In this study, the cross‐amplification of a commercial multiplex set of 11 cattle (Bos taurus) microsatellites was tested on a panel of 35 European bison (Bison bonasus) individuals. After polymerase chain reaction optimization, all loci cross‐amplified successfully in investigated bisons. Number of alleles and observed and expected heterozygosity per locus are in the range of 2–4, 0.086–0.629 and 0.288–0.621 respectively. The availability of a heterologous set of multiplexed microsatellite markers derived from cattle opens an avenue for collecting profound genetic data for efficient conservation management strategies of the European bison.     

种群遗传变异及基因多样度分析

阐述了种群遗传学和生态遗传学研究的基本单位－种群的定义和意念含义,其一是种系变异含义,其二是生态适应含义,并概述了遗传变异的研究历史。从３个方面及质量性状变异,数量性状变异与基金变异,介绍了种群遗传变异的研究方法,重点介绍了基因变异的测量和亚种群基因多样度的测量,并且依据研究所得的遗传参数对中国苜蓿不同种群遗传结构和多样度进行了例证分析。     

Exploring the diversity and molecular evolution of shrews (family Soricidae) using mtDNA cytochrome b data

The taxonomy of the shrews (family Soricidae) is primarily underpinned by the analysis of morphology characters. These morphological characters are, however, often plesiomorphic and difficult to interpret in an evolutionarily meaningful way. Recent molecular studies have provided some clarity at lower-taxonomic levels, but studies giving resolution to deeper levels of the soricid phylogeny are lacking. By constructing the most taxonomically inclusive molecular phylogeny for this cosmopolitan family, this study aims to test the utility of the routinely used mitochondrial cytochrome b gene to resolve the higher-level relationships within the Soricidae. Given the life history characteristics of shrews (high metabolic rate in Soricinae, small body size, fast generation time and generally short life span), the metabolic theory predicts accelerated nucleotide turnover in this family. By examining the molecular evolution of the cytochrome b molecule, this study identifies both lineage- and codon-specific mutational rate differences.We suggest that codon-specific mutational rate patterns are best explained by unique selective regimes operating on the different codon positions, while lineage-specific rate variation among genera belonging to Soricidae may reflect differences in the mode and speed of speciation events which generated species.     

Molecular cloning,organization, expression and 3D structural analysis of the MHC class Ia gene in the whitespotted bamboo shark (Chiloscyllium plagiosum)

Cartilaginous fishes are the oldest jawed vertebrates, from which the major histocompatibility complex (MHC) derived approximately 500 MYA; however, full-length genomic sequences for MHC genes in these species remain undescribed. This lack of basic information about MHC organization in cartilaginous fish is hindering investigations into the relationship between MHC polymorphism and disease, and leaves a large gap in our understanding of shark MHC evolution. Here, we obtained a complete 4887 bp genomic DNA of chplUAA (designated as chplUAA) from the whitespotted bamboo shark (Chiloscyllium plagiosum) using long PCR. The full-length cDNA sequence was 1385 bp, with a 1029 bp open reading frame (ORF) encoding 343 amino acids. Six unique sequences (chplUAA*01-06) were detected from 51 sequences from three samples. No more than two sequences were found in each individual, suggesting that only one UAA locus was amplified in each sample. Phylogenetic analysis supports monophyly of all available shark classical class Ia sequences.     

Molecular cloning, expression pattern, and 3D structural analysis of the MHC class IIB gene in the Chinese longsnout catfish (Leiocassis longirostris)

Major histocompatibility complex (MHC) class I and class II molecules encode glycoproteins which mediate the specificity of the vertebrate adaptive immune response. In this study, MHC class IIB gene from the Chinese longsnout catfish (Leiocassis longirostris) was cloned and sequenced, which encoded a predicted protein of 248 amino acids (28.06 kDa) containing a signal peptide, a beta 1 domain, a beta 2 domain, a connecting peptide, a transmembrane region, and a cytoplasmic tail. Using PCR with primers designed from known fish MHC class IIB sequences followed by elongation of the 5' and 3' ends using rapid amplification of cDNA ends (RACE), the full-length cDNA of longsnout catfish MHC class IIB was identified to be 1293 bp, consisting of a 26 bp 5'-terminal untranslated region (UTR), a 520 bp 3'-UTR, and a 747 bp open reading frame (ORF) bearing characteristics of the immunoglobulin C-type 1 (IGc1) family. The deduced amino acid sequences of the Chinese longsnout catfish MHC class IIB gene had 58-75% identity with those of other fishes. Six class IIB alleles were identified from five individuals. At most two different alleles observed in each individual may infer the existence of a single locus of class IIB gene in the Chinese longsnout catfish genome. An extensive study of polymorphism was examined in 60 individuals. A total of 11 haplotypes of exon 2 were detected in the sampled Chinese longsnout catfish. The rates of nonsynonymous substitutions (d(N)) occurred at a higher frequency than that of synonymous substitutions (d(S)), suggesting the polymorphism of exon 2 seemed to be maintained by the balancing selection. By using long PCR technique, the genomic sequence was further identified to be 2345 bp in length, which contained six exons and five introns. Interestingly, a 98 bp intron 5 cut the 3'-UTR into two parts. Real-time quantitative RT-PCR demonstrated high expression of MHC IIB in gills, spleen, head kidney, and intestine, moderate expression in liver and stomach, and low or negligible expression in heart. Homology modelling has been applied to create a 3D model of longsnout catfish MHC class IIB, with features evaluated and discussed to investigate its interaction with CD4 participating in antigen recognition. The present findings will provide valuable information for understanding structure, function, expression, and molecular polymorphism of MHC class IIB in adaptive immunity of the Chinese longsnout catfish and teleost.     

基于微卫星标记的中国斑翅山鹑遗传多样性及遗传结构分析

斑翅山鹑(Perdix dauuricae)是具有很高经济价值的猎用禽.为了获得其群体遗传多样性及遗传变异信息,为我国斑翅山鹑进一步有效保护和合理利用提供科学依据,本研究共采用了8个微卫星标记位点对分布于我国北方的斑翅山鹑23个地理种群285个个体进行了群体遗传多样性及遗传结构分析.结果表明,我国斑翅山鹑为遗传多样性较丰富的群体,且每一群体均表现出较高的遗传多样性,其中柴达木盆地地区的种群拥有最高的遗传多样性.遗传分化分析显示组间及组内种群间均表现为遗传分化显著,遗传分化系数(FST)值表明大部分种群间表现出显著的遗传分化.系统树构建和贝叶斯聚类分析结果均显示斑翅山鹑23个地理种群被分化为明显的两个组群.另外,通过BOTTLENECK对各地理种群的分析显示斑翅山鹁种群曾经历过近期瓶颈效应.本研究结果表明,柴达木盆地地区种群、华北平原地区种群、静宁种群、张家川种群以及六盘山地区种群应予以相应关注,以保证有足够的种群大小来保持这些种群的遗传多样性.     

cDNA cloning, genomic structure and expression analysis of the goose (Anser cygnoides) MHC class I gene

To provide data for studies on avian disease resistance, goose MHC class I cDNA (Ancy-MHC I) was cloned from a goose cDNA library, it's genomic structure and expression analysis were investigated. The mature peptides of Ancy-MHC I cDNA encoded 333 amino acids. The genomic organization is composed of eight exons and seven introns. Based on the genetic distance, six Ancy-MHC I genes from six individuals can be classified into four lineages. A total of nineteen amino acid positions in peptide-binding domain showed high scores by Wu-kabat index analysis. The Ancy-MHC I amino acid sequence displayed seven critical HLA-A2 amino acids that bind with antigen polypeptides, and have an 85.4-98.9% amino acid homology with each genes, and a 59.8-66.0% amino acid homology with chicken MHC class I. Expression analyses using Q-RT-PCR to detect the tissue-specific expression of Ancy-MHC I mRNA in an adult goose. The result appeared that Ancy-MHC I cDNA was expressed in the liver, spleen, intestine, kidney, lung, pancreas, heart, brain, and skin. The phylogenetic tree appears to branch in an order consistent with accepted evolutionary pathways.     

Molecular cloning, polymorphism and tissue distribution of the MHC class IIB gene in the Chinese goose (Anser cygnoides)

1. The goose major histocompatibility complex (MHC) class IIB cDNA (Ancy-MHCII) was cloned by homology cloning and rapid amplification of cDNA ends by polymerase chain reaction (RACE-PCR), and the genomic structure and tissue expression were investigated. 2. Three different 5'-RACE sequences (Ancy-MHC II5'-1, Ancy-MHC II5'-2, Ancy-MHC II5'-3), one 3'-RACE sequence (Ancy-MHC II-3') and two different full length Ancy-MHC IIB cDNA sequences (Ancy-CD01, Ancy-CD02), which came from different alleles at one locus or different loci, were determined. 3. The genomic organisation is composed of 6 exons and 5 introns, with a longer intron region than that of the chicken. The alleles encode 259 and 260 amino acids in the mature protein. 4. The number of non-synonymous substitutions (dN) in the peptide-binding region of exon 2 from 8 alleles was higher than that of the synonymous substitutions (dS). 5. Tissue-specific expression of Ancy-MHC II mRNA was detected in an adult goose using RT-PCR. These results showed that Ancy-MHC II mRNA was expressed in the lung, spleen, liver, intestine, heart, kidney, pancreas, brain, skin and muscle. This is consistent with the expression of MHC class IIB in various tissues from the chicken. 6. Sequences from goose, snipe and duck clustered together when compared with known MHC class IIB sequences from the other species, significantly differing from mammals and aquatic species, indicating a pattern consistent with accepted evolutionary pathways.     

Spontaneous Aujeszky's disease (pseudorabies) in European wild boars (Sus scrofa) in the federal state of Brandenburg, Germany

In this report two cases of spontaneous Aujeszky's disease (AD, syn. pseudorabies) in European wild boars (Sus scrofa), in the federal state of Brandenburg, Germany, are described. Both animals displayed severe central nervous disturbances, including loss of fear of humans, disorientation, and tremors of head and limbs, and were shot by hunters for laboratory exclusion of rabies. The main finding in the well nourished, juvenile (approx. 7-8 months) animals was a non-suppurative panencephalitis characterized by neuronal necroses, intranucelar eosinophilic inclusion bodies in necrotic neurons, spongiosis of the neuropil, gliosis and perivascular cuffing of lymphocytes, plasma cells and eosinophilic granulocytes. Pseudorabies virus (PrV)-antigen was detected by immunohistochemistry in typically affected brain areas and was isolated from pooled tissues (brain and tonsil) in both cases. The molecular characterization of the virus isolates revealed that they belonged to the wild boar-associated PrV subtype Iw. These cases indicate that spontaneous AD can sporadically occur in free living wild boars under natural conditions. However, factors triggering the disease, e. g. social stress, age related change from passive to active immunity, individual susceptibility to PrV infection and environmental conditions, have to be clarified by future experimental studies.     

The tortoises (Testudinidae) and terrapins (Pelomedusidae) of southern Africa: their diversity,distribution and conservation

Southern Africa has the richest diversity of land tortoises in the world, as well as an important radiation of pelomedusid terrapins. Total species richness has two epicentres, including the Transvaal lowveld and adjacent KwaZulu/Natal (owing to the prevalence of pelomedusid terrapins) and the Eastern and south-western Cape (owing to small testudinids). The area encompassing Lesotho, Transkei and adjacent regions, lacks testudinids for unknown reasons. Archaeological data indicates that this gap is natural, and not the result of man-induced extinctions. Endemic species are clustered in the Cape, whilst the few threatened species are more widely distributed. The majority of species is well protected in existing reserves. The small number of chelonian species in southern Africa and their relatively well-known distributions, test the efficacy of an iterative reserve selection algorithm. The presence of many allopatric (or nearly so) congeneric species leads to the selection of iterative reserves that protect peripheral populations. To avoid this, marginal records and isolated, peripheral populations should be excluded from the analysis.     

Population genetic structure and diversity of the East Balkan Swine (Sus scrofa) in Bulgaria,revealed by mitochondrial DNA and microsatellite analyses

The East Balkan Swine (EBS) is the only indigenous pig breed in Bulgaria. We analyzed the mitochondrial DNA (mtDNA) control region and 21 microsatellite loci for 198 individuals from 11 farms in Bulgaria. Obtained 11 mtDNA haplotypes including three novel ones were grouped to two major clades, European clade E1 (146/198 individuals, 73.7%) and Asian clade A (52/198, 26.3%). The mixture of the two clades may have resulted from historical crossbreeding between the European and Asian pig breeds. Clade A was frequent in southeastern Bulgaria (Burgas Province), but less frequent or absent in northeastern Bulgaria (Varna and Shumen Provinces). The distribution of Europe- and Asia-specific haplotypes relative to EBS farm locations could be attributed to regional differences of breeding systems (e.g., crossbreeding with imported commercial pigs). A microsatellite analysis showed high heterozygosities for all the EBS farms, and negative inbreeding coefficients presumably due to crossing with commercial pigs or wild boars and/or efforts to reduce inbreeding by farmers. Bayesian clustering analyses showed that all farm populations are genetically well distinguishable from one another. Although diversity has been maintained by the efforts of farmers and a breeding association, the effective population size remains small, and conservation efforts should be continued.     

New insights into the genetic diversity of European porcine reproductive and respiratory syndrome virus (PRRSV)

The complete ORF5 sequences of 66 porcine reproductive and respiratory syndrome (PRRS) field virus strains (1991-2001) and three European modified live vaccine strains were determined, as well as ORFs 6 and 7 of 19 selected strains. The variability of the deduced ORF5 amino acid sequences was analysed using statistical process control (SPC), allowing for the objective assessment of variable and conserved regions. Four variable and four conserved regions as well as five hypervariable amino acid positions were defined. The effects of genetic variability on possible structural and functional properties were discussed with emphasis on immunogenic features. Phylogenetic analysis and pairwise comparison of the nucleotide sequences revealed that the genetic distances between the strains has greatly increased over time. The data do not support an evolutionary influence of the geographical location or the time of sample collection, nor of PRRSV vaccination on strain development. In contrast to other authors who tended to concentrate on the samples from either a common geographic origin or a short sampling period, we could not confirm geographically separate PRRSV clusters nor did we find evidence of positive selective pressure as measured by the ratio of synonymous to non-synonymous substitutions in ORF5, 6 or 7. Immunological implications and vaccination strategies are discussed.     

东方白鹳微卫星遗传标记的筛选及其在种群遗传分析中的应用

东方白鹳(Ciconia boyciana)是分布于东亚的大型濒危涉禽,亟待深入开展其种群的保护遗传学研究.本研究采用微卫星标记技术筛选东方白鹤的微卫星位点,并进行种群的遗传多样性分析.所用的36对微卫星引物7对来自白鹳(Ciconia ciconia),12对来自朱鹮(Nipponia nippon),17对来自大蓝...     

The European College of Veterinary Pathologists (ECVP): the professional body for European veterinary pathologists

The European College of Veterinary Pathologists (ECVP) was established in 1995 with the aim of advancing veterinary pathology and promoting high standards within the specialty in Europe. The ECVP is one of 21 European colleges recognized by the European Board of Veterinary Specialisation (EBVS), which represents a quality-assurance system for European veterinary specialists. Until the ECVP was founded, there was no unified European system recognizing the specialty of pathology, and many European countries followed their own qualification systems, which varied in form and standard. The ECVP provides an annual certifying examination, the passing of which is required to gain membership (diplomate status) in the college. This qualification is now accepted on equal terms by the well-established American College of Veterinary Pathologists (ACVP). In line with EBVS requirements, the ECVP has also established a standard continuing professional development (CPD) and re-registration system for its membership. Furthermore, it has promoted and unified European post-graduate training in veterinary pathology by setting up requirements for residency training programs and making registration and monitoring of these programs by the ECVP a prerequisite for approval of an institution as a training facility. The concurrent establishment, together with the European Society of Veterinary Pathology, of an annual summer school that trains residents for the certifying examination has further fostered European post-graduate training. Within 10 years, the ECVP has succeeded in establishing common standards and a unified approach to veterinary pathology throughout Europe. This article describes the evolution and organization of the ECVP.     

Genetic diversity,phylogenetic position and morphometric analysis of Astacus colchicus (Decapoda,Astacidae): a new insight into Eastern European crayfish fauna

The phylogeny of European crayfish fauna, especially with respect to Eastern European species, is still far from being completely resolved. To fill this gap, we analyzed most of the European crayfish species focusing on the phylogenetic position of the endemic crayfish Astacus colchicus, inhabiting Georgia. Three mitochondrial and one nuclear marker were used to study evolutionary relationships among European crayfish species, resulting in the unique phylogenetic position of A. colchicus indicating independent species status to A. astacus. Phylogenetic analyses revealed a deep molecular divergence of A. colchicus in comparison to A. astacus (6.5–10.9% in mtDNA and 1.1% in nDNA) as well as to Pontastacus leptodactylus and P. pachypus (5.5–10.0% in mtDNA and 1.4–2.4% in nDNA). Absent ventral process on second male pleopod and abdominal somites II and III with pleura rounded lacking prominent spines clearly indicate taxonomic assignment to the genus Astacus; however, the species is distributed almost in the middle of Ponto-Caspian area typical by occurrence of the genus Pontastacus. Several morphological indices linked to head length, carapace, and total body length and width were found to demonstrate apparent differences between A. colchicus and A. astacus. Although this study provides a novel insight into European crayfish phylogeography, we also point out the gaps in comprehensive study of the P. leptodactylus species complex, which could reveal details about the potential species status of particular species and subspecies within this genus.