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Dermatophytoses caused by Microsporum canis are frequently encountered in cats and dogs; they are highly contagious and readily transmissible to humans. In this study, two single genes, respectively coding for dipeptidyl peptidases IV and V (DppIV and DppV), were isolated and characterized. Both proteins share homology with serine proteases of the S9 family, some of which display properties compatible with implication in pathogenic processes. Both genes are expressed in vivo in experimentally infected guinea-pigs and in naturally infected cats, and when the fungus is grown on extracellular matrix proteins as the sole nitrogen and carbon source. DppIV and V were produced as active recombinant proteases in the yeast Pichia pastoris; the apparent molecular weight of rDppV is 83 kDa, whereas rDppIV appears as a doublet of 95 and 98 kDa. Like other members of its enzymatic subfamily, rDppIV has an unusual ability to cleave Pro-X bonds. This activity does not enhance the solubilization of keratin by fungal secreted endoproteases, and the protease probably acts solely on small soluble peptides. RDppV showed no ability to induce delayed-type hypersensitivity (DTH) skin reactions in guinea-pigs, despite the known immunogenic properties of homologous proteins. [ABSTRACT FROM AUTHOR]

This is the first documented case report of dermatitis associated with the poultry mite ( Dermanyssus gallinae) in a horse. It occurred in a 16-year-old horse that was in contact with domestic hens. Clinical signs consisted of severe pruritus, with self-induced hair loss mainly on the head. Despite the multiple skin scrapings performed during both day- and nighttime, mites were only isolated from the in-contact poultry and from the horse's environment, and not the horse. The animal was treated using a 2% permethrin solution, sprayed on the entire body once a week for 4 weeks, and by decontamination of the horse's immediate environment. Although eradication of the mites and elimination of further contact between the horse and the poultry were not achievable, recurrence of dermatitis was prevented by regular applications of permethrin on the horse and biannual decontamination of the horse's stable. [ABSTRACT FROM AUTHOR]

Dermatomycoses caused by Microsporum canis are frequent in domestic animals and easily transmissible to humans. Several proteases secreted by this fungus were identified as potential virulence factors, but the construction of deficient strains is required to investigate their role in the pathogenesis of the disease. Using target genes encoding two of these proteases, a first evaluation of the utility of RNA-mediated silencing as a reverse genetic tool in dermatophytes was carried out. SUB3 and DPPIV, respectively coding for a subtilisin and a dipeptidyl peptidase, were both down-regulated, by means of two plasmid constructs designed to express an RNA hairpin that corresponds to part of their respective sequence. The degree of attenuation was evaluated by enzymatic assay of the transformants culture supernatants, and by real-time reverse transcriptase-polymerase chain reaction. Enzymatic activities and expression levels varied from less than 5% to 100% of that of control transformants obtained with plasmid without hairpin inserts. Inhibition was globally more efficient for SUB3 than for DPPIV. These results show that RNA silencing can be used for functional genomics in M. canis, and particularly to circumvent the limits and technical difficulties of conventional disruption methods. [ABSTRACT FROM AUTHOR]

In order to identify protective immunogens against Microsporum canis infection, a purified recombinant keratinolytic metalloprotease (r-MEP3) was tested as a subunit vaccine in experimentally infected guinea pigs. Both humoral and cellular specific immune responses developing towards r-MEP3 were evaluated, by enzyme-linked immunosorbent assay and by in vitro lymphocyte transformation tests respectively. Vaccination induced a strong antibody response, and a significant but transient lymphoproliferative response against the protein. However, the protocol failed to prevent fungal invasion or development of dermatophytic lesions. These results show that under the present experimental conditions, r-MEP3 specific antibodies are not protective against a challenge exposure. They also suggest that in the same model, the induction of cell-mediated immunity towards r-MEP3 is not sufficient, indicating the need for further research in the field of specific immune mechanisms involved in M. canis dermatophytosis. [Copyright &y& Elsevier]

In order to better understand the host-fungus relationship in Microsporum canis dermatophytosis and to identify major fungal antigens, the immune response to a crude exoantigen preparation and to a purified recombinant keratinolytic metalloprotease (r-MEP3) was evaluated in guinea pigs experimentally infected with M. canis . Humoral and cellular immune responses were assessed from day 0 to day 57 post-infection (PI), the former by enzyme-linked immunosorbent assay (ELISA) and the latter via a lymphocyte proliferation assay. Infected guinea pigs developed humoral and cellular responses to both M. canis exoantigen and r-MEP3, while no specific immune response to these antigens was observed in control animals. This is the first report on the development of both humoral and cell-mediated immune responses to a purified keratinase in M. canis dermatophytosis. [ABSTRACT FROM AUTHOR]

A secreted 31.5-kDa keratinolytic subtilase (SUB3; AJ431180) is thought to be a Microsporum canis virulence factor and represents a candidate for vaccination trials. In this study, the recombinant keratinase (r-SUB3) was produced by the Pichia pastoris expression system and purified to homogeneity. Recombinant SUB3 displayed identical biochemical properties with the native protease. Experimentally cutaneously infected guinea pigs showed specific lymphoproliferative response towards r-SUB3, while no specific humoral immune response was induced except for one animal. The heterologous expression of SUB3 provides a valuable tool for addressing further investigations on the role of this keratinase in the specific cellular immune response and on its use in vaccination trials in the cat. [Copyright &y& Elsevier]

A letter to the editor is presented about the breeding sites of bluetongue vectors in northern Europe. [ABSTRACT FROM AUTHOR]

Microsporum canis is a worldwide distributed zoophilic and zoonotic dermatophyte which is responsible for most cases of feline ringworm. Its prevalence is on the rise in developed countries. Since 1995, the author and his Laboratory have developed a research programme to better understand the pathogenesis of M. canis dermatophytosis. Both the potential fungal virulence factors and the host immune response were investigated. A 31.5 kDa keratinolytic subtilase (SUB) and a 43.5 kDa keratinolytic metalloprotease (MEP) were purified from a crude exoantigen consisted in a mixture of inducible proteins secreted by the fungus in a medium enriched with keratin. Additionally, two gene families encoding respectively 3 SUBs and 3 MEPs, including the 2 keratinases previously purified (called SUB3 and MEP3), were characterized. The genes encoding several MEPs and SUBs were shown to be transcribed in vivo during infection by nested RT-PCR. Finally, we demonstrated that a specific cellular immune response towards the crude exoantigen, MEP3 and SUB3 was induced in experimentally infected guinea pigs. This indicates that the potential immunoprotective properties of these antigens are interesting to evaluate in the frame of the development of an efficient vaccine against feline ringworm.