Twelve toll-like receptor (TLR) genes in the family Equidae - comparative genomics, selection and evolution.

Toll-like receptors (TLRs) represent an important part of the innate immune system. While human and murine TLRs have been intensively studied, little is known about TLRs in non-model species. The order Perissodactyla comprises a variety of free-living and domesticated species exposed to different pathogens in different habitats and is therefore suitable for analyzing the diversity and evolution of immunity-related genes. We analyzed TLR genes in the order Perissodactyla with a focus on the family Equidae. Twelve TLRs were identified by bioinformatic analyses of online genomic resources; their sequences were confirmed in equids by genomic DNA re-sequencing of a panel of nine species. The expression of TLR11 and TLR12 was confirmed in the domestic horse by cDNA sequencing. Phylogenetic reconstruction of the TLR gene family in Perissodactyla identified six sub-families. TLR4 clustered together with TLR5; the TLR1-6-10 subfamily showed a high degree of sequence identity. The average estimated evolutionary divergence of all twelve TLRs studied was 0.3% among the Equidae; the most divergent CDS were those of Equus caballus and Equus hemionus kulan (1.34%) in the TLR3, and Equus africanus somaliensis and Equus quagga antiquorum (2.1%) in the TLR1 protein. In each TLR gene, there were haplotypes shared between equid species, most extensively in TLR3 and TLR9 CDS, and TLR6 amino acid sequence. All twelve TLR genes were under strong negative overall selection. Signatures of diversifying selection in specific codon sites were detected in all TLRs except TLR8. Differences in the selection patterns between virus-sensing and non-viral TLRs were observed.

Associations between the presence of specific antibodies to the West Nile Virus infection and candidate genes in Romanian horses from the Danube delta.

The West Nile virus (WNV) is a mosquito-borne flavivirus causing meningoencephalitis in humans and animals. Due to their particular susceptibility to WNV infection, horses serve as a sentinel species. In a population of Romanian semi-feral horses living in the Danube delta region, we have analyzed the distribution of candidate polymorphic genetic markers between anti WNV-IgG seropositive and seronegative horses. Thirty-six SNPs located in 28 immunity-related genes and 26 microsatellites located in the MHC and LY49 complex genomic regions were genotyped in 57 seropositive and 32 seronegative horses. The most significant association (pcorr < 0.0002) was found for genotypes composed of markers of the SLC11A1 and TLR4 genes. Markers of five other candidate genes (ADAM17, CXCR3, IL12A, MAVS, TNFA), along with 5 MHC class I and LY49-linked microsatellites were also associated with the WNV antibody status in this model horse population. The OAS1 gene, previously associated with WNV-induced clinical disease, was not associated with the presence of anti-WNV antibodies.

Genetic susceptibility to West Nile virus infection in Camargue horses.

West Nile virus (WNV) is a mosquito-borne zoonotic neurotropic virus capable to cause lethal meningoencephalitis (WNE) in infected hosts such as birds, horses, and humans. Due to their sensitivity, horses serve as sentinel species in areas at risk. We studied a population of Camargue horses living in Southern France in two zones with endemic WNV circulation where WNV outbreaks were recorded in 2000 and 2003-4. Two sets of microsatellite markers located in MHC and Ly49 genomic regions were genotyped as well as multiple SNPs in ten immunity-related candidate gene regions. Associations between genetic polymorphisms and resistance/susceptibility to WNE were tested. While single marker associations were weak, compound two-gene genotypes of SNPs located within the MAVS, NCR2 and IL-10 genes and microsatellites HMS082 and CZM013 were associated with susceptibility to WNE. Combinations of microsatellite markers CZM009, ABGe17402 and ABGe9019 were associated with simple seroconversion without clinical signs of WNE (resistance). In addition, a distribution of polymorphic markers between WNV-IgG seropositive horses and a control group of WNV-IgG seronegative horses was tested. One SNP in the OAS1 gene (NC_009151.3:g.21961328A>G) was significantly associated with the seropositive phenotype (pcorr = 0.023; OR = 40.5 CI (4.28; 383.26); RR = 8.18 CI (1.27; 52.89) in the Camargue breed. In compound genotypes, SNP markers for SLC11A1, MAVS, OAS1, TLR4, ADAM17 and NCR2 genes and ten microsatellites showed non-random distribution between seropositive and seronegative groups of horses. Further analysis of associated markers could contribute to our understanding of anti-WNV defense mechanisms in horses.

Microsatellite markers for evaluating the diversity of the natural killer complex and major histocompatibility complex genomic regions in domestic horses.

Genotyping microsatellite markers represents a standard, relatively easy, and inexpensive method of assessing genetic diversity of complex genomic regions in various animal species, such as the major histocompatibility complex (MHC) and/or natural killer cell receptor (NKR) genes. MHC-linked microsatellite markers have been identified and some of them were used for characterizing MHC polymorphism in various species, including horses. However, most of those were MHC class II markers, while MHC class I and III sub-regions were less well covered. No tools for studying genetic diversity of NKR complex genomic regions are available in horses. Therefore, the aims of this work were to establish a panel of markers suitable for analyzing genetic diversity of the natural killer complex (NKC), and to develop additional microsatellite markers of the MHC class I and class III genomic sub-regions in horses. Nine polymorphic microsatellite loci were newly identified in the equine NKC. Along with two previously reported microsatellites flanking this region, they constituted a panel of 11 loci allowing to characterize genetic variation in this functionally important part of the horse genome. Four newly described MHC class I/III-linked markers were added to 11 known microsatellites to establish a panel of 15 MHC markers with a better coverage of the class I and class III sub-regions. Major characteristics of the two panels produced on a group of 65 horses of 13 breeds and on five Przewalski's horses showed that they do reflect genetic variation within the horse species.

Haematological and biochemical variations among eight sighthound breeds.

OBJECTIVE: The aim of the study was to compare the haematological and biochemical profiles of eight sighthound breeds. DESIGN: Samples were taken from 192 individuals of the sighthound breeds (Whippet, Greyhound, Italian Greyhound, Sloughi, Saluki, Borzoi, Pharaoh Hound and Azawakh). Routine haematological and biochemical examinations were performed and the results were evaluated statistically. RESULTS: There were significant differences in haematology and clinical biochemistry among the sighthound breeds. The most similar laboratory profile to Greyhounds was found in Whippets. Italian Greyhounds had significantly higher alanine aminotransferase activity than other sighthounds, except Pharaoh Hounds. CONCLUSION: Application of the Greyhound laboratory profile to other sighthounds is not recommended because of the frequent differences in haematological and clinical biochemical reference intervals.

Determinants of the prevalence of Heligmosomum costellatum (Heligmosomidae: Trichostrongyloidea) in a common vole population in southern Moravia, Czech Republic.

A study of nematode (Heligmosomum costellatum) prevalence in the common vole (Microtus arvalis) population was performed in 2002 and 2003 in an agricultural region of southern Moravia, Czech Republic. The influence of the trapping date, body weight, age, gender and reproduction of the common vole on the prevalence of the larvae of H. costellatum encapsulated in the stomach wall was examined. Of 503 common voles examined, 27.6% were H. costellatum positive. The prevalence of H. costellatum in the common vole population was influenced mainly by the weight, trapping date, year and reproductive state, as well as by the interactions between year and gender and between year and reproductive state. The influence of age was weak. The most infected common vole groups were the older, heavier and already reproducing females captured from April to August and all voles captured in 2002. In general, parasite prevalence seems to be dependent on the population parameters of the host species. However, the H. costellatum prevalence study in a single common vole population was not sufficient to fully describe and explain the nematode-host interactions.

[Effect of a prolonged stay in space on the reaction of the adrenal cortical and medullary layers]. / Vliianie dlitel'nogo prebyvaniia v kosmose na reaktsiiu korkovogo i mozgovogo sloia nadpochechnikov.

Concentrations of corticosteron and catecholamines, and activities of catecholamine synthesizing enzymes, tyrosine hydroxylase, dopamine-beta-hydroxylase and phenyl ethanol amine-N-methyl-transferase, were measured in the adrenals of rats flown for 19.5 days aboard the biosatellite Cosmos-782, synchronous and vivarium control animals sacrificed on R + 0 and R + 26 days. The flown rats showed a moderate but significant increase in the adrenal weight, corticosteron concentration and tyrosine hydroxylase activity on R+0 day. These parameters returned to the normal on R+26 day. This data gives evidence that the adrenocortical and sympathoadrenal systems were not significantly stimulated by the space flight effects; therefore, weightlessness cannot be considered as a potent stressful agent, although its some stressogenic effect has been documented.