Taylorella equigenitalis is the causative agent of sexually transmitted contagious equine metritis. Infections manifest as cervicitis, vaginitis and endometritis and cause temporary infertility and miscarriages of mares. While previous studies have analyzed this organism for various parameters, the evolutionary dynamics of this pathogen, including the emergence of antibiotic resistance, remains unresolved. The aim of this study was to isolate contemporary strains, determine their genome sequences, evaluate their antibiotic resistance and compare them with other strains. We determined nine complete whole genome sequences of T. equigenitalis strains, mainly from samples collected from Kladruber horses in the Czech Republic. While T. equigenitalis strains from Kladruby isolated between 1982 and 2018 were inhibited by streptomycin, contemporary strains were found to be resistant to streptomycin, suggesting the recent emergence of this mutation. In addition, we used the collection dates of Kladruber horse strains to estimate the genome substitution rate, which resulted in a scaled mean evolutionary rate of 6.9×10-7 substitutions per site per year. Analysis with other available T. equigenitalis genome sequences (n = 18) revealed similarity of the Czech T. equigenitalis genomes with the Austrian T. equigenitalis genome, and molecular dating suggested a common ancestor of all analyzed T. equigenitalis strains from 1.5-2.6 thousand years ago, dating to the first centuries A.D. Our study revealed a recently emerged streptomycin resistance in T. equigenitalis strains from Kladruber horses, emphasizing the need for antibiotic surveillance and alternative treatments. Additionally, our findings provided insights into the pathogen's evolution rate, which is important for understanding its evolution and preparing preventive strategies.
- MeSH
- Anti-Bacterial Agents pharmacology MeSH
- Drug Resistance, Bacterial genetics MeSH
- Phylogeny MeSH
- Genome, Bacterial * genetics MeSH
- Horses microbiology MeSH
- Evolution, Molecular MeSH
- Horse Diseases * microbiology MeSH
- Whole Genome Sequencing * MeSH
- Taylorella equigenitalis * genetics MeSH
- Animals MeSH
- Check Tag
- Female MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Geographicals
- Czech Republic MeSH
Equine neosporosis is an intracellular protozoan disease with a global distribution, affecting a diverse range of warm-blooded animals. Neospora caninum Dubey, Carpenter, Speer, Topper et Uggla, 1988 is associated with foetal loss, neurological disease and abortion in equids. No information was available regarding equine N. caninum infection among equids in Iraq. Thus, the aim of this study was to determine the prevalence rate of N. caninum in equines by using a competitive enzyme-linked immunosorbent assay (c-ELISA). A total of 329 blood samples randomly selected from equines, comprising 268 horses and 61 donkeys were examined. The seroprevalence rate of N. caninum was determined as 46% (28/61) for donkeys and 24% (64/268) for horses. The prevalence of N. caninum indicated a significantly higher risk of infection in donkeys compared to horses (P < 0.001). However, the odds of N. caninum infection in draught equids were 8.2 times greater than other equids with a significant difference (P < 0.001). The current study revealed no significant differences in the prevalence of N. caninum across various genders, breeds, clinical statuses, disease histories and among equids that had contact with dogs. While outdoor feeding and mixed (grazing), showed a significant difference (P = 0.003) and (P = 0.75), respectively, in the presence of antibodies against N. caninum compared to indoor feeding (stable). Moreover, the odds of infection in equids with a history of late abortion were 4.8 times higher than those without such a history of abortion (2.20-10.56) with statistical significance (P < 0.001).
- MeSH
- Enzyme-Linked Immunosorbent Assay * veterinary MeSH
- Equidae * parasitology MeSH
- Coccidiosis * veterinary epidemiology parasitology MeSH
- Horses MeSH
- Horse Diseases * epidemiology parasitology MeSH
- Neospora * isolation & purification MeSH
- Prevalence MeSH
- Antibodies, Protozoan blood MeSH
- Seroepidemiologic Studies MeSH
- Animals MeSH
- Check Tag
- Male MeSH
- Female MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Geographicals
- Iraq MeSH
- MeSH
- Chemoprevention methods MeSH
- Dermatologic Agents therapeutic use MeSH
- Adult MeSH
- Wound Healing * MeSH
- Horses MeSH
- Insect Bites and Stings * drug therapy therapy MeSH
- Post-Exposure Prophylaxis methods MeSH
- Rabies Vaccines therapeutic use MeSH
- Treatment Outcome MeSH
- Check Tag
- Adult MeSH
- Male MeSH
- Publication type
- Case Reports MeSH
- Geographicals
- Tunisia MeSH
Equids may be infected by zoonotic Leishmania spp., including Leishmania infantum, in regions where canine leishmaniasis (CanL) is endemic, and Leishmania martiniquensis, which has been reported in horses from Central Europe. This study was designed to evaluate the occurrence of both Leishmania spp. among equids living in CanL endemic areas of Italy, as well as to identify dipteran vectors from the same habitats. From March to October 2023, blood, serum and tissue samples from skin lesions were collected from equids (n = 98; n = 56 donkeys and n = 42 horses) living in Italy, as well as sand flies and biting midges. Blood samples (n = 98) and skin lesions (n = 56) were tested for Leishmania spp. by conventional and real time PCRs and sera were tested by immunofluorescence antibody tests (IFAT) for both L. infantum and L. martiniquensis. Insects were morphologically identified, and female specimens (n = 268 sand flies, n = 7 biting midges) analyzed for Leishmania DNA, as well as engorged sand flies (n = 16) for blood-meal detection. Two animals with skin lesions (i.e., one donkey and one horse) scored positive for Leishmania spp. DNA, and 19 animals (i.e., 19.4%; n = 13 donkeys and n = 6 horses) were seropositive for L. infantum, with five of them also for L. martiniquensis. Most seropositive animals had no dermatological lesions (i.e., 68.4%) while both animals molecularly positive for Leishmania spp. scored seronegative. Of the 356 sand flies collected, 12 females (i.e., n = 8 Sergentomyia minuta; n = 3 Phlebotomus perniciosus, n = 1 Phlebotomus perfiliewi) were positive for Leishmania spp. DNA, and one out of seven biting midges collected was DNA-positive for L. infantum. Moreover, engorged sand flies scored positive for human and equine DNA. Data suggest that equids living in CanL endemic areas are exposed to Leishmania spp., but their role in the circulation of the parasite needs further investigations.
- MeSH
- Ceratopogonidae parasitology MeSH
- Endemic Diseases veterinary MeSH
- Equidae * parasitology MeSH
- Insect Vectors * parasitology MeSH
- Horses parasitology MeSH
- Leishmania infantum isolation & purification genetics MeSH
- Leishmania * isolation & purification genetics classification MeSH
- Leishmaniasis * veterinary epidemiology parasitology transmission MeSH
- Horse Diseases parasitology epidemiology MeSH
- Dog Diseases * parasitology epidemiology transmission MeSH
- Dogs MeSH
- Psychodidae parasitology MeSH
- Animals MeSH
- Check Tag
- Male MeSH
- Dogs MeSH
- Female MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Geographicals
- Italy MeSH
- MeSH
- Chemoprevention methods MeSH
- Dermatologic Agents therapeutic use MeSH
- Adult MeSH
- Wound Healing * MeSH
- Horses MeSH
- Bites and Stings * therapy MeSH
- Humans MeSH
- Skin Care methods MeSH
- Post-Exposure Prophylaxis methods MeSH
- Rabies Vaccines therapeutic use MeSH
- Check Tag
- Adult MeSH
- Humans MeSH
- Male MeSH
- Publication type
- Case Reports MeSH
- Geographicals
- Tunisia MeSH
The immunogenome is the part of the genome that underlies immune mechanisms and evolves under various selective pressures. Two complex regions of the immunogenome, major histocompatibility complex (MHC) and natural killer cell receptor (NKR) genes, play an important role in the response to selective pressures of pathogens. Their importance is expressed by their genetic polymorphism at the molecular level, and their diversity associated with different types of diseases at the population level. Findings of associations between specific combinations of MHC/NKR haplotypes with different diseases in model species suggest that these gene complexes did not evolve independently. No such associations have been described in horses so far. The aim of the study was to detect associations between MHC and NKR gene/microsatellite haplotypes in three horse breed groups (Camargue, African, and Romanian) by statistical methods; chi-square test, Fisher's exact test, Pearson's goodness-of-fit test and logistic regression. Associations were detected for both MHC/NKR genes and microsatellites; the most significant associations were found between the most variable KLRA3 gene and the EQCA-1 or EQCA-2 genes. This finding supports the assumption that the KLRA3 is an important receptor for MHC I and that interactions of these molecules play important roles in the horse immunity and reproduction. Despite some limitations of the study such as low numbers of horses or lack of knowledge of the selected genes functions, the results were consistent across different statistical methods and remained significant even after overconservative Bonferroni corrections. We therefore consider them biologically plausible.
- MeSH
- Antilymphocyte Serum * therapeutic use MeSH
- Anemia, Aplastic * drug therapy MeSH
- Cyclosporine * therapeutic use MeSH
- Child MeSH
- Immunosuppressive Agents * therapeutic use MeSH
- Infant MeSH
- Horses MeSH
- Rabbits MeSH
- Humans MeSH
- Adolescent MeSH
- Child, Preschool MeSH
- Retrospective Studies MeSH
- Treatment Outcome MeSH
- Animals MeSH
- Check Tag
- Child MeSH
- Infant MeSH
- Rabbits MeSH
- Humans MeSH
- Adolescent MeSH
- Male MeSH
- Child, Preschool MeSH
- Female MeSH
- Animals MeSH
- Publication type
- Letter MeSH
- Comparative Study MeSH
Innumerable similarities in reproductive cyclicity and hormonal alterations highlight the considerable utility of the mare to study aspects of follicular dynamics and reproductive function in view of the largely constricted, human research subjects. The bi-directional communication between the growing oocyte and the surrounding somatic cells embodies the hallmark of mammalian follicular development, partially mediated by extracellular vesicles (EVs) encapsulated with microRNAs (miRNAs) and present in the follicular fluid (FF). Here, we aimed to decipher the dynamics of the miRNAs in EVs from equine FF aspirated in vivo during different stages of follicular development, namely, predeviation (PreDev; 18-20 mm), deviation (Dev; 22-25 mm), postdeviation (PostDev; 26-29 mm), preovulatory (PreOV; 30-35 mm), and impending ovulation (IMP; ∼40 mm). Approximately 176 known miRNAs were found in all groups with 144 mutually detected among all groups. Cluster analysis exhibited 15 different expression patterns during follicular development. Among these patterns, a group of 22 miRNAs (including miR-146b-5p, miR-140, and miR-143) exhibited a sharp reduction in expression from the PreDev until the PreOV stage. Another cluster of 23 miRNAs (including miR-106b, miR-199a-5p, and miR-125a-5p) exhibited a stable expression pattern at the PreDev stage until the PostDev stage, with a significant increase at the PreOV stage followed by a significant decrease at the IMP stage. In conclusion, this study provides greater insights into the stage-specific expression dynamics of FF EV-miRNAs during equine follicular development, which may propose novel approaches to improve ART and provide new biomarkers to facilitate the assessment of ovarian pathophysiological conditions.
- MeSH
- Extracellular Vesicles * genetics metabolism MeSH
- Follicular Fluid metabolism MeSH
- Horses MeSH
- Humans MeSH
- MicroRNAs * metabolism MeSH
- Ovarian Follicle metabolism MeSH
- Ovulation genetics MeSH
- Mammals MeSH
- Animals MeSH
- Check Tag
- Humans MeSH
- Female MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Research Support, N.I.H., Extramural MeSH
The major histocompatibility complex (MHC) with its class I and II genes plays a crucial role in the immune response to pathogens by presenting oligopeptide antigens to various immune response effector cells. In order to counteract the vast variability of infectious agents, MHC class I and II genes usually retain high levels of SNPs mainly concentrated in the exons encoding the antigen binding sites. The aim of the study was to reveal new variability of selected MHC genes with a special focus on MHC class I physical haplotypes. Long-range NGS to was used to identify exon 2-exon 3 alleles in three genetically distinct horse breeds. A total of 116 allelic variants were found in the MHC class I genes Eqca-1, Eqca-2, Eqca-7 and Eqca-Ψ, 112 of which were novel. The MHC class II DRA locus was confirmed to comprise five exon 2 alleles, and no new sequences were observed. Additional variability in terms of 15 novel exon 2 alleles was identified in the DQA1 locus. Extensive overall variability across the entire MHC region was confirmed by an analysis of MHC-linked microsatellite loci. Both diversifying and purifying selection were detected within the MHC class I and II loci analyzed.
- MeSH
- Alleles MeSH
- Exons genetics MeSH
- Genes, MHC Class II * MeSH
- Major Histocompatibility Complex MeSH
- Horses genetics MeSH
- Histocompatibility Antigens Class I * MeSH
- Histocompatibility Antigens Class II genetics MeSH
- Binding Sites MeSH
- Animals MeSH
- Check Tag
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
Propidium iodide (PI) and YO-PRO-1 (YPI) dyes are routinely used to determine sperm viability in many livestock species. It is commonly accepted that these dyes penetrate only sperm cells with damaged plasma membranes. Recently, however, the mechanism of dye uptake unrelated to damaged plasma membranes, but instead related to pannexin channels in dog and stallion sperm cells was demonstrated. This pilot study aimed to evaluate the role of pannexins in the uptake of PI and YPI dyes on Wallachian frozen-thawed ram spermatozoa by flow cytometry using probenecid, a specific inhibitor of pannexin channels. Additionally, the expression of pannexins in Wallachian sperm was evaluated directly (by qRT-PCR). The results demonstrate the active role of pannexin channels in the uptake of PI and YPI dyes on frozen-thawed Wallachian ram sperm. In conclusion, when using the PI or YPI exclusion assay to determine Wallachian frozen-thawed ram sperm viability, the danger of overestimating the number of spermatozoa with the damaged plasma membrane must be considered. The observed breed-specific, and more importantly, individual differences in gene expression as well as in dye uptake indicate the need for further studies.
- MeSH
- Coloring Agents MeSH
- Benzoxazoles MeSH
- Quinolinium Compounds * MeSH
- Iodides * MeSH
- Horses MeSH
- Cryopreservation methods veterinary MeSH
- Pilot Projects MeSH
- Propidium MeSH
- Dogs MeSH
- Semen MeSH
- Spermatozoa MeSH
- Semen Preservation * veterinary methods MeSH
- Animals MeSH
- Check Tag
- Male MeSH
- Dogs MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
