Clinicopathological and pedigree investigation of a novel spinocerebellar neurological disease in juvenile Quarter Horses in North America.

BACKGROUND: In 2020, a novel neurologic disease was observed in juvenile Quarter Horses (QHs) in North America. It was unknown if this was an aberrant manifestation of another previously described neurological disorder in foals, such as equine neuroaxonal dystrophy/equine degenerative myeloencephalopathy (eNAD/EDM). HYPOTHESIS/OBJECTIVES: To describe the clinical findings, outcomes, and postmortem changes with Equine Juvenile Spinocerebellar Ataxia (EJSCA), differentiate the disease from other similar neurological disorders, and determine a mode of inheritance. ANIMALS: Twelve neurologically affected QH foals and the dams. METHODS: Genomic DNA was isolated and pedigrees were manually constructed. RESULTS: All foals (n = 12/12) had a history of acute onset of neurological deficits with no history of trauma. Neurological deficits were characterized by asymmetrical spinal ataxia, with pelvic limbs more severely affected than thoracic limbs. Clinicopathological abnormalities included high serum activity of gamma-glutamyl transferase and hyperglycemia. All foals became recumbent (median, 3 days: [0-18 days]), which necessitated humane euthanasia (n = 11/12, 92%; the remaining case was found dead). Histological evaluation at postmortem revealed dilated myelin sheaths and digestion chambers within the spinal cord, most prominently in the dorsal spinocerebellar tracts. Pedigree analysis revealed a likely autosomal recessive mode of inheritance. CONCLUSIONS AND CLINICAL IMPORTANCE: EJSCA is a uniformly fatal, rapidly progressive, likely autosomal recessive neurological disease of QHs <1 month of age in North America that is etiologically distinct from other clinically similar neurological disorders. Once the causative variant for EJSCA is validated, carriers can be identified through genetic testing to inform breeding decisions.

NetF-producing Clostridium perfringens and its associated diseases in dogs and foals.

The role of type A Clostridium perfringens in canine acute hemorrhagic diarrhea syndrome and foal necrotizing enteritis is poorly characterized. However, a highly significant association between the presence of novel toxigenic C. perfringens and these specific enteric diseases has been described. These novel toxigenic strains produce 3 novel putative toxins, which have been designated NetE, NetF, and NetG. Although not conclusively demonstrated, current evidence suggests that NetF is likely the major virulence factor in strains responsible for canine acute hemorrhagic diarrhea syndrome and foal necrotizing enteritis. NetF is a beta-pore-forming toxin that belongs to the same toxin superfamily as CPB and NetB toxins produced by C. perfringens. The netF gene is encoded on a conjugative plasmid that, in the case of netF, also carries another putative toxin gene, netE. In addition, these strains consistently also carry a cpe tcp-conjugative plasmid, and a proportion also carry a separate netG tcp-conjugative plasmid. The netF and netG genes form part of a locus with all the features of the pathogenicity loci of tcp-conjugative plasmids. The netF-positive isolates are clonal in origin and fall into 2 clades. Disease in dogs or foals can be associated with either clade. Thus, these are strains with unique virulence-associated characteristics associated with serious and sometimes fatal cases of important enteric diseases in 2 animal species.

Reversible dysphagia secondary to guttural pouch mycosis in a gelding treated medically with voriconazole and surgically with carotid occlusion and esophagostomy.

A gelding was diagnosed with dysphagia and left guttural pouch mycosis. Treatments included topical antifungal drugs, systemic voriconazole, and balloon occlusion of the internal carotid artery. Ongoing dysphagia of neurological origin necessitated extra-oral feeding through an esophagostomy tube. Complementary case management included acupuncture. Clinical remission occurred over 10 weeks.

Dysphagie réversible secondaire à une mycose de la poche gutturale chez un hongre traité médicalement avec du voriconazole et chirurgicalement par l'occlusion de la carotide et l'œsophagostomie. Un hongre a été diagnostiqué avec de la dysphagie et une mycose de la poche gutturale gauche. Les traitements ont inclus des médicaments antifongiques topiques, du voriconazole systémique et l'occlusion par ballon de l'artère carotide interne. Une dysphagie non résorbée d'origine neurologique a nécessité une alimentation extra-orale par un tube d'œsophagostomie. Une gestion du cas complémentaire a inclus l'acupuncture. Une rémission clinique s'est produite pendant 10 semaines.(Traduit par Isabelle Vallières).

Necrotizing hepatitis associated with Clostridium novyi in a pony in western Canada.

Severe icterus, peritoneal effusion, localized fibrinous peritonitis, and necrotizing hepatitis were found at necropsy of a 20-year-old female pony with a history of acute onset depression, inappetence, fever, and marked elevation in hepatic enzymes. Gross pathology, histopathology, and immunohistochemistry were compatible with a diagnosis of clostridial hepatitis caused by Clostridium novyi-group bacteria. This is believed to be the first reported case of clostridial hepatitis in an equid in Canada, and only the third report of this rare disease in North America.

Hépatite nécrosante associée àClostridium novyichez un poney de l'Ouest canadien. Un ictère grave, une effusion péritonéale, une péritonite fibrineuse localisée et une hépatite nécrosante ont été constatées chez un poney femelle âgé de 20 ans avec une anamnèse d'apparition soudaine de dépression, d'inappétence, de fièvre et d'élévations marquées des enzymes hépatiques. La pathologie clinique, l'histopathologie et l'immunohistochimie étaient compatibles avec un diagnostic d'hépatite clostridiale causée par une bactérie du groupe Clostridium novyi. On croit qu'il s'agit du premier cas signalé d'hépatite clostridiale chez un équidé au Canada et seulement le troisième rapport de cette maladie rare en Amérique du Nord.(Traduit par Isabelle Vallières).

NetF-producing Clostridium perfringens: Clonality and plasmid pathogenicity loci analysis.

Clostridium perfringens is an important cause of foal necrotizing enteritis and canine acute hemorrhagic diarrhea. A major virulence determinant of the strains associated with these diseases appears to be a beta-sheet pore-forming toxin, NetF, encoded within a pathogenicity locus (NetF locus) on a large tcp-conjugative plasmid. Strains producing NetF also produce the putative toxin NetE, encoded within the same pathogenicity locus, as well as CPE enterotoxin and CPB2 on a second plasmid, and sometimes the putative toxin NetG within a pathogenicity locus (NetG locus) on another separate large conjugative plasmid. Previous genome sequences of two netF-positive C. perfringens showed that they both shared three similar plasmids, including the NetF/NetE and CPE/CPB2 toxins-encoding plasmids mentioned above and a putative bacteriocin-encoding plasmid. The main purpose of this study was to determine whether all NetF-producing strains share this common plasmid profile and whether their distinct NetF and CPE pathogenicity loci are conserved. To answer this question, 15 equine and 15 canine netF-positive isolates of C. perfringens were sequenced using Illumina Hiseq2000 technology. In addition, the clonal relationships among the NetF-producing strains were evaluated by core genome multilocus sequence typing (cgMLST). The data obtained showed that all NetF-producing strains have a common plasmid profile and that the defined pathogenicity loci on the plasmids are conserved in all these strains. cgMLST analysis showed that the NetF-producing C. perfringens strains belong to two distinct clonal complexes. The pNetG plasmid was absent from isolates of one of the clonal complexes, and there were minor but consistent differences in the NetF/NetE and CPE/CPB2 plasmids between the two clonal complexes.

Plasmid Characterization and Chromosome Analysis of Two netF+ Clostridium perfringens Isolates Associated with Foal and Canine Necrotizing Enteritis.

The recent discovery of a novel beta-pore-forming toxin, NetF, which is strongly associated with canine and foal necrotizing enteritis should improve our understanding of the role of type A Clostridium perfringens associated disease in these animals. The current study presents the complete genome sequence of two netF-positive strains, JFP55 and JFP838, which were recovered from cases of foal necrotizing enteritis and canine hemorrhagic gastroenteritis, respectively. Genome sequencing was done using Single Molecule, Real-Time (SMRT) technology-PacBio and Illumina Hiseq2000. The JFP55 and JFP838 genomes include a single 3.34 Mb and 3.53 Mb chromosome, respectively, and both genomes include five circular plasmids. Plasmid annotation revealed that three plasmids were shared by the two newly sequenced genomes, including a NetF/NetE toxins-encoding tcp-conjugative plasmid, a CPE/CPB2 toxins-encoding tcp-conjugative plasmid and a putative bacteriocin-encoding plasmid. The putative beta-pore-forming toxin genes, netF, netE and netG, were located in unique pathogenicity loci on tcp-conjugative plasmids. The C. perfringens JFP55 chromosome carries 2,825 protein-coding genes whereas the chromosome of JFP838 contains 3,014 protein-encoding genes. Comparison of these two chromosomes with three available reference C. perfringens chromosome sequences identified 48 (~247 kb) and 81 (~430 kb) regions unique to JFP55 and JFP838, respectively. Some of these divergent genomic regions in both chromosomes are phage- and plasmid-related segments. Sixteen of these unique chromosomal regions (~69 kb) were shared between the two isolates. Five of these shared regions formed a mosaic of plasmid-integrated segments, suggesting that these elements were acquired early in a clonal lineage of netF-positive C. perfringens strains. These results provide significant insight into the basis of canine and foal necrotizing enteritis and are the first to demonstrate that netF resides on a large and unique plasmid-encoded locus.

Development of a live, attenuated, potential vaccine strain of R. equi expressing vapA and the virR operon, and virulence assessment in the mouse.

Pneumonia caused by Rhodococcus equi remains a significant problem in foals. The objective of this study was to develop a safe and efficacious attenuated strain of R. equi for eventual use in oral immunization of foals. The approach involved expression of vapA in a live, virulence plasmid-negative, strain of R. equi (strain 103-). PCR-amplified fragments of the vapA gene, with and without the upstream genes virR, orf5, vapH, orf7 and orf8 (orf4-8), were cloned into a shuttle vector pNBV1. These plasmids, named pAW48A and pAWVapA respectively, were electroporated into strain 103-. The presence of the recombinant vectors in the attenuated strain (103-) and the integrity of the inserted genes were confirmed, and both constructs expressed VapA. The virulence of the two strains was compared to that of wild type R. equi 103+ and negative controls by their intravenous inoculation into mice, followed by examination of liver clearance 4 days later. Mice inoculated with R. equi 103-, 103-/pAWVapA and 103-/pNBV1 completely cleared infection, whereas strain 103-/pAW48A persisted in 47% of mice.

Foaling rates and risk factors for abortion in pregnant mares presented for medical or surgical treatment of colic: 153 cases (1993-2005).

The purpose of this study was to determine foaling rates in mares presented for medical or surgical treatment of colic, and to examine risk factors associated with abortion following colic. A retrospective analysis of 153 medical records found that mares treated surgically for colic (P = 0.0007) were 3.5 times more likely to have a negative pregnancy outcome than were mares treated medically for colic. Anesthetic time (P = 0.01) and intra-operative hypotension (P = 0.03) were significantly associated with negative pregnancy outcome. Mares with an anesthetic time > or = 3 h were 6 times more likely to abort. Signs of endotoxemia (P = 0.30), hypoxia (P = 0.89), flunixin meglumine administration (P = 0.13), mucous membrane color at the time of presentation (P = 0.82) and capillary refill time (P = 0.76) were not associated with pregnancy outcome. There was no difference in the foaling rate for mares that had received progestin supplementation versus those that had not (P = 0.42). In this study, the significant risk factors for abortion were surgically treated colic, long anesthetic time, and intraoperative hypotension.