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Operationalizing MCI criteria changes the relative frequency of MCI subtypes. Nonamnestic single domain MCI may be a previously nonrecognized type of MCI associated with SVD.

Streptococcus (S.) dysgalactiae subsp. equisimilis is responsible for severe diseases in humans, including primary bacteraemia, pneumonia, endocarditis, and toxic shock syndrome. Infection in some animal species can also occur, although a few studies have looked into cross-species infectivity. In horses, S. equisimilis is generally considered infrequent or opportunistic, but has recently been isolated from cases of strangles-like disease. Rapid and sensitive diagnostic techniques could enable epidemiological studies and effective investigation of outbreaks involving these bacteria. In this study, PCR protocols previously described in cattle and in humans to detect the species S. dysgalactiae and the subspecies equisimilis were evaluated to detect specific sequences in equine samples. For this purpose, 99 monolateral nasal swabs were collected from horses from stud farms with a history of S. equisimilis infection and were tested blindly by bacteriological isolation and by single and duplex PCR. DNA for PCR was extracted both from the colonies grown on agar media and from enrichment broth aliquots after incubation with nasal swab samples. S. equisimilis was identified by bacteriological isolation in 23 out of 99 swab samples, and PCR assays on these colonies were fully concordant with bacteriological identification (kappa statistic = 1.00). In addition, PCR of the enrichment broth aliquots confirmed the bacteriological results and detected S. equisimilis in 6 samples more than the bacteriological examination (kappa statistic = 0.84). The PCR protocols appeared to be reliable for the rapid identification of S. equisimilis in equine nasal swab samples, and could be useful for microbiological diagnosis.

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We report the expression and characterization of the putative catalytic subunit (pORF30) and accessory protein (pORF18) of equine herpesvirus 1 DNA polymerase, which are encoded by open reading frames 30 and 18 and are homologous to herpes simplex virus type 1 UL30 and UL42, respectively. In vitro transcriptiontranslation of open reading frames 30 and 18 generated proteins of 136 and 45 kDa, respectively. In vitroexpressed pORF30 possessed basal DNA polymerase activity that was stimulated by pORF18, as measured by DNA polymerase assays in vitro. Purified baculovirus-expressed pORF30 exhibited DNA polymerase activity similar to that of the in vitro-expressed protein, and baculovirus-expressed pORF18 could stimulate both nucleotide incorporation and long-chain DNA synthesis by pORF30 in a dose-and time-dependent manner. The salt optima for activity of both pORF30 and the holoenzyme were substantially different from those for other herpesvirus DNA polymerases. As demonstrated by yeast two-hybrid assays, pORF30 and pORF18 could physically interact, most likely with a 1:1 stoichiometry. Finally, by mutational analysis of the 1,220-residue pORF30, we demonstrated that the extreme C terminus of pORF30 is important for physical and functional interaction with the accessory protein, as reported for UL30 and other herpesvirus DNA polymerases. In addition, a C-proximal region of pORF30, corresponding to residues 1114 to 1172, is involved in binding to, and stimulation by, pORF18. Taken together, the results indicate that pORF30 and pORF18 are the equine herpesvirus 1 counterparts of herpes simplex virus type 1 UL30 and UL42 and share many, but not all, of their characteristics.Equine herpesvirus 1 (EHV-1) is a pathogen of major importance in horses that can induce a wide spectrum of diseases (1, 45). Like other herpesviruses, EHV-1 establishes a lifelong infection, via a quiescent state known as latency. The outcomes of infection, either following primary infection or reactivation from latency, vary from mild/unapparent respiratory disease to the induction of abortion in pregnant mares and, in extreme cases, neurological disease resulting in paralysis and ultimately death.EHV-1 is classified on biological grounds as a member of the Alphaherpesvirinae, a subfamily of the herpesviruses which is typified by herpes simplex virus type 1 (HSV-1) and also includes varicella-zoster virus (VZV) and pseudorabies virus (PRV) (49). The complete DNA sequence of a pathogenic British isolate (strain Ab4) of EHV-1 was recently determined (56). The genome is approximately 150 kbp in size, is similar in arrangement to the HSV-1, VZV, and PRV genomes, and contains 80 open reading frames (ORFs), corresponding to 76 distinct genes, likely to encode proteins. Comparison of predicted amino acid sequences to those of HSV-1, VZV, and PRV homologs allowed the functions of many EHV-1 proteins to be putatively assigned. However, only a few of these proteins have been studied so far, although the number of characterized proteins is rapidly increasing....