The Immune Response to Microsporum canis Induced by a Fungal Cell Wall Vaccine*

Abstract— An experimental infection model was used to assess induction of specific immunity against Microsporum canis in cats with an M. canis cell wall vaccine preparation. Kittens 8–9 weeks old (n= 12) received five doses of either vaccine or placebo at biweekly intervals. Specific immunity was monitored via plasma anti-dermatophyte antibody titers and lymphocyte blastogenesis (LB) to dermatophyte antigens. After vaccination, cats were challenged with viable M. canis spores, and lesion development was monitored. Vaccinated cats developed higher anti-dermatophyte IgG, but not IgM, titers than controls, beginning after the second dose of vaccine (P < 0.001). During the vaccination period, specific cellular immunity as measured by LB was absent in control cats, but developed to a limited degree in vaccinated cats (P < 0.05). After challenge with 10⁵ fungal spores per cat, both control and vaccinated cats developed active infections. The vaccine appeared to induce an antibody titer quantitatively similar to that produced by infection, but less measured cellular immunity than was seen with infection and recovery. These results suggest that induction of high titers of serum IgG or IgM antibody against Microsporum canis is not protective against challenge exposure.     

Ringworm in dogs and cats

Ringworm is an uncommon disease of dogs, but cats, especially the longhaired breeds, are more frequently infected. Of the several dermatophytes involved, Microsporum canis causes 94 per cent of ringworm in cats and 65 per cent in dogs. This species is highly contagious for man but the true incidence of human infection is unknown. Ringworm caused by M canis usually responds readily to treatment, but when the infection establishes in a colony of cats, eradication can be difficult and expensive. Other forms of ringworm in dogs and cats are caused by dermatophytes acquired from wild animals, mainly small rodents such as mice and voles. This is an uncommon and trivial disease of cats, but some infections in dogs can be remarkably persistent and difficult to resolve.     

Epidemiology and Clinical Features of Dermatophytosis in Dogs and Cats at Louisiana State University: 1981–1990

Abstract— Dermatophytes were cultured from seventy of 1824 (3.8 per cent) canine samples and 61 of 408 (14.9 per cent) feline samples submitted over ten years. Microsporum canis was the most common species isolated (86/131). M. gypseum (31/70) and M. canis (30/70) were isolated most frequently from dogs whereas M. canis was isolated most frequently from cats (56/61). Both male and female dogs and cats were equally affected by dermatophytosis. There was a higher incidence in both dogs and cats less than one year of age. Mixed breed dogs (19/70), domestic shorthaired cats (39/61) and Persian cats (15/61) were most often affected. M. gypseum had a greater incidence of infection in summer and most often caused localized disease in dogs. In dogs, localized dermatophytosis was more common. Localized infections were most often caused by M. canis in all animals, with the head (23/75), feet (10/75) and tail (7/75) affected most frequently. Résumé— Dos dermatophytes ont été isolés à partir de 70/1824 (3,8 pourcent) prélèvements canins et 61/408 (14,9 pourcent) prélèvements félins collectés sur une période de dix ans. L'espèce la plus souvent isolée était Microsporum canis (86/131). Microspomm gypseum (31/70) et M. canis (30/70) étainet les plus fréquemment isolés chez les chiens, alors qu'il s'agissait de M. canis (56/61) chez les chats. Les mâles et les femelles, chiens ou chats, étaient également représentés. Il y avait une nette incidence des animaux de moins d'un an, tant chez les chiens que chez les chats. Les chiens croisés (19/70), les chats européens (39/61) et les chats Persans (15/61) étaient les plus souvent atteints. L'incidence de M. gypseumétait plus importante l'été et provoquait les plus souvent des lésions localisées les chiens. Les formes localisées étaient plus fréquentes chez les chiens et le plus souvent dues àM. canis dans les deux espèces, avec une atteinte préférentielle de la tête (23/75), des membres (10/75) et de la queue (7/75). Zausammenfassung— Aus siebzig von 1824 Proben von Hunden (3,8 Prozent) und 61 von 408 Proben von Katzen (14,9 Prozent) von einem Zeitraum über zehn Jahre wurden Dermatophyten angezüchtet. Microsporum canis war die am häufigsten isolierte Spezies (86/131). Microsporum gypseum (31/70) und M. canis (30/70) wurden am häufigsten bei Hunden isoliert, M. canis dagegen wurde am häufigsten bei Katzen isoliert (56/61). Männliche wie weibliche Hunde und Katzen waren gleichermaßen von Dermatomykosen betroffen. Sowohl bei Hunden also auch bei Katzen war die Inzidenz bei Tieren unter einem Jahr höher. Am häufigsten erkrankten Mischlingshunde (19/70), Kurzhaarhauskatzen (39/61) und Perserkatzen (15/61). Die Inzidenz der Infektion mit M. gypseum war im Sommer größer, bei Hunden verursachte es mehr lokalisierte Erkrankungen. Bei Hunden überhaupt waren lokalisierte Dermatomykosen häufiger anzutreffen. Lokalisierte Infektionen wurden bei alien Tieren am häufigsten durch M. canis verursacht, am meisten betroffen waren Kopf (23/75), Pfoten (10/75) und Schwanz (7/75). Resumen Se cultivaron dermatofitos de setenta de 1824 (3,8 por ciento) muestras caninas y 61 de 408 (14,9 cor ciento) muestras felinas, sometidas durante diez anos. Microsporum canis fue la mas común de las especies aisladas (86/131). Microsporum gypseum (31/70) y M. canis (30/70) se aislaron mas frecuentemente de perros, mientras que M. canis fue aislado con más frecuencia de gatos (56/61). Ambos, macho y hembra, perros y gatos fueron igualmente afectados por la dermatofítosis. Había una mayor incidencia en perros y gatos menores de un año de edad. Perros de cruce de razas (19/70), gatos domésticados de pelo corto (39/61) y gatos persas (15/61) fueron afectados con más frecuencia. M. gypseum tenía una mayor incidencia de infección en verano, y muy frecuentemente causaba enfermedades localizados en perros. En perros, fue más commún la dermatofitosis localizada. Infecciones localizadas eran más frecuentemente causadas por M. canis en todos los animates, con la cabeza (23/75), pies (10/75) y rabo (7/75) afectados con más frecuencia.     

The Value of Enzyme-linked Immunosorbent Assay (ELISA) in the Sero-diagnosis of Canine Dermatophytosis Due to Microsporum Canis

Abstract— Fourteen sera from dogs naturally or experimentally infected with Microsporum canis were examined by enzyme-linked immunosorbent assay (ELISA) in order to study the humoral response to infection. A group of 12 animals (7 laboratory dogs and 5 dogs from the natural environment) was used as controls. ELISA optical density (OD) values in twofold serial dilutions from 1:100 to 1:51200 were compared between the groups. Significant antibody response to infection with M. canis was observed in naturally and experimentally infected animals and confirmed by threshold determination. Specificity for the dermatophyte fungi was tested by performing cross-reactivity assays with three fungal antigens, Malassezia pachydermatis, Candida albicans and Aspergillus fumigatus. There were no statistically significant indications of cross-reactions, except with anti-A. fumigatus IgG, where differences were noticed within the groups of naturally infected animals and controls. The use of ELISA technique in sero-diagnosis of dermatophytosis in dogs is discussed. Résumé— 14 sérums chiens infectés naturellement ou expérimentalement par Microsporum canis ont été testés par une technique Immunoenzymatique (ELISA) pour étudier la réponse immunitaire humorale à cette infection. Le groupe témoin était composé de 12 animaux (7 chiens de laboratoire et 5 chiens d'un milieu naturel). Les valeurs de densité optique (DO) obtenues avec des dilutions sériées de raison 2 des sérums (1:100 à 1:51,500) ont été comparées entre chaque groupe. Des taux significativement élevés d'anticorps spécifiques de Microsporum canis ont été retrouvés chez les animaux infectés naturellement et expérimentalement et confirmés par l'établissement d'un seull de positivité. La spécificité de la technique vis-à-vis des dermatophytes a été appréciée en effectuant des tests d'antigénicité croisée avec des Malassezia pachydermatis, Candida albicans et Aspergillus fumigatus. II n'y avait pas de réactions croisées significatives, exception faite des IgG spécifiques d'A. fumigatus, pour lequel il existait des differences de réponse entre les animaux infectés naturellement et le groups témoin. L'utilisation d'une technique ELISA dans le diagnostic des dermatophyties canines est discutées. Zusammenfassung— Vierzehn Seren von Hunden, die natürlich oder experimentell mit Microsporum canis infiziert waren, wurde mit dem ELISA-Verfahren (Enzyme Linked Immunosorbent Assay) untersucht, um die humorale Antwort auf die lnfektion zu verfolgen. Eine Gruppe von 12 Tieren (7 Laborhunde und 5 Hunde aus normaler Haltung) dienten als Kontrolle. ELISA-optische Dichtewerte (OD) in zweifacher serieller Verdünnung von 1:100 bis 1:51200 wurden zwischen beiden Gruppen verglichen. Bei den natürlich und experimentell infizierten Tieren konnte eine signifikante Antikörperreaktion auf die lnfektion mit M. canis beobachtet und durch Schwellenwertbestimmung bestätigt werden. Die Spezifität für Dermatophyten wurde überprüft, indem Kreuzreaktionsproben mit drei Pilzantigenen, Malassezia pachydermatis, Canidida albicans und Aspergillus fumigatus, durchgeführt wurden. Statistisch signifikante Anzeichen für Kreuzreaktionen konnten nicht nachgewiesen werden, außer bei Anti-A. fumigatus-IgG, bei dem Unterschiede innerhalb der Gruppe der natürlich infizierten und der Kontrolltiere beobachtet wurden. Es wird über die Brauchbarkeit der ELISA-Technik für die Serodiagnose von Dermatophyten bei Hunden diskutiert. Resumen Se examinaron un total de 14 sueros procedentes de perros infectados de forma natural y artificial con Microsporum canis por el método de enzima ligado de inmunoabsorbencia, (ELISA), con el fin de estudiar la respuesta humoral a la infección. Como controles, se usaron un grupo de 12 animales (7 perros de laboratorio y 5 perros que vivían en condiciones naturales). Los valores de densidad óptica ELISA, (OD), fueron comparados entre los grupos en dos diluciones seriadas de 1:100 a 1:51,200. Una respuesta significativa de anticuerpos a la infección con Microsporum canis fue observada en perros infectados de forma natural y artificial, y confirmada por la determinación del punto crítico. La especificidad del dermatofito füngico se estudió por medio de ensayos de reactividad cruzada con otros tres antigenos fúngicos, Malassezia pachydermatis, Candida albicans y Aspergillus fumigatus. No se encontraron indicaciones de reacciones cruzadas estadísticamente significativas, excepto con la IgG de A. fumigatus, donde se advirtieron diferencias en el group de animales infectados naturalmente y en el grupo control. El uso de la téchnica ELISA en el diagnóstico serólogico de las dermatofilosis en el perro, es discutido.     

Microsporum canis: Inapparent carriage by cats and the viability of arthrospores

A total of 181 dermatologically healthy pet cats from 177 different households, attending a veterinary clinic, were sampled for the presence of dermatophytes by a modified MacKenzie hair brush technique. Microsporum canis was the only dermatophyte recovered and was isolated from four cats (2.2 per cent) from four different households. In addition to clinical details, owners were questioned about the environment and management of all the cats sampled. The data regarding the cats from which M canis was recovered showed little variation from that of the culture-negative cats except that all four cats were from multi-cat (more than two cats) households, whereas only 35 per cent of the culture-negative cats were from a similar environment. The viability of M canis in infected feline hairs stored at room temperature was maintained for between 13 and 18 months.     

P‐10 Comparison of carpet and toothbrush methods for the detection of asymptomatic carriage of dermatophytes in cats

Diagnosis of Microsporum canis infection is a challenge in cats with suspected asymptomatic carriage. The aim of this study was to compare the carpet method and the toothbrush method of sample collection for dermatophyte culture. The study was conducted on apparently healthy cats in a chronically infected cattery. Sampling was performed with both a sterilized piece of carpet and a toothbrush applied to the entire body surface. Samples were inoculated on Sabouraud's media (with chloramphenicol and cycloheximide). Multiple (6) applications with the toothbrush were necessary to obtain a similar surface area of inoculation. Cultures were incubated at 27°C, then examined from day 5 to 21 for the presence and number of colonies of M. canis and other fungi. Some samples were inoculated twice (initially negative or with a low number of M.canis). A total of 112 cultures were performed (14 duplicates) from 44 cats. On day 21, infection was detected in 23 cats (54.7%): 20 (87%) with the carpet method and 21 (91%) with the toothbrush method. None of the replicated cultures allowed the detection of new cases but three were subsequently negative. In eight cases, only one to six colonies of M. canis were obtained. Both methods used are easy to perform on cats and have a similar sensitivity of approximately 90%. The carpet method is, however, much less expensive, easier to prepare, mail, store and inoculate. The lack of growth in some duplicate cultures (initial development of one or two colonies) may explain the limits of diagnosis of infection and justify the need for multiple inoculations of agar with the toothbrush method. Funding: Self‐funded.     

Immunological Reactivity to Intradermal Dermatophyte Antigens in Cats with Dermatophytosis

Abstract— To study the immune response of cats to dermatophyte infection, ten healthy cats and five cats previously infected with Microsporum canis were injected intradermally with dermatophyte glycoprotein extracts. Injection sites were observed for immediate and delayed reactions and biopsied at 24 and 48 hours after injection. All five previously-infected cats but only two of 10 control cats had immediate positive reactions to M. canis antigen. Four out of five previously-infected cats and one of 10 control cats also had reactions to Trichophyton rubrum antigen, suggesting antigenic crossreactivity between these fungal species. At 24 h after injection, four out of five previously-infected cats had delayed reactions to M. canis antigen at the injection sites; equivalent sites in all control animals were negative. The histological appearance of the delayed reactions was a superficial perivascular dermatitis, with lymphohistiocytic cells, eosinophils, mast cells, and neutrophils. This pattern is consistent with a delayed-type hypersensitivity response to fungal antigens. Special stains revealed no evidence of cutaneous basophil hypersensitivity in these reactions. Our data suggest that naturally-occurring feline dermatophytosis is accompanied by development of both humoral and cellular immune responses to the organism. Intradermal skin testing with dermatophyte antigens is a useful tool for investigating the immune response of cats to dermatophytosis. Résumé— Afin d'étudier la réponse immunitaire des chats à une infection par des dermatophytes, des intradermoréactions à l'aide d'extraits glycoprotéiques de dermatophytes ont été pratiquées sur 10 chats sains et 5 chats antérieurement infectés par Microsporum canis. Les sites d'injection ont cté observés pour lés réactions immédiates et retardées et biopsiés à 24 et 48 heures. Les 5 chats antéricurement infectés et seulement 2 des chtas témoins ont présenté des réactions immédiates à l'extrait antigénique de Microsporum canis, 4 des 5 chats antérieurement infectés et 1 des 10 chats témoins ont aussi présenté des réactions àTríchophyton rubrum, suggérant l'existence de réactions croisées entre ces deux espèces de dermatophytes. 24 heures après l'injection, 4 des 5 chats antérieurement infectés ont présenté des réactions retardées àMicrosporum canis au point d'injection. Ces mêmes tests étaient négatifs chez les animaux témoins. L'aspect histologique des réactions retardées etait celui d'une dermite périvasculaire superficielle à celles lymphohistiocylaires, éosinophiles, neutrophiles, et mastocytes. Cet aspect est compatibles avec une réaction d'hypersensibilité retardée aux antigènes fongiques. Des colorations spécifiques n'ont pas permis de mettre en évidence une hypersensibilité cutanée à basophiles dans ces réactions. Ces résultats suggèrent que l'infection naturelle par des dermatophytes chez le chat entraine le développement d'une réaction immunitaire humorale et cellulaire spécifique de ces organismes. Les intradermoréactions à l'aide d'extraits antigéniques de dermatophytes sont un bon moyen d'exploraton de la réponse immunitaire des chats lors de dermatophytle. Zausammenfassung— Um die Immunantwort von Katzen auf die Infektion mit Dermatophyten zu untersuchen, wurden bei zehn gesunden Katzen und fünf Tieren mit vorausgegangener Microsporum canis-Infektion Extrakte von Dermatophyten-Glykoprotein intradermal injiziert. Die Injektionsstellen wurden auf Sofortund Spätreaktionen kontrolliert sowie 24 und 48 Stunden nach der Injektion biopsiert. Alle fünf Katzen mit vorausgegangener Infektion, aber nur zwei der 10 Kontrolltiere zeigten eine positive Sofortreaktion auf M. canis-Antigen. Vier der fünf früher infizierten Katzen und eine der 10 Kontrollkatzen zeigten gleichzeitig Reaktionen auf Tríchophyton rumbrum-Anúgen. Dies legt den Verdacht nahe, daß zwischen diesen Pilzspezies Kreuzreaktionen auf Antigene auftreten. 24 Stunden nach der Injektion zeigten vier der fünf früher infizierten Katzen Spätreaktionen auf M. canis-Antigen an den Injektionsstellen. Entsprechende Hautstellen waren bei alien Kontrolltieren negativ. Das histologische Bild der Spätreaktionen bestand in einer oberflächlichen, perivaskulären Dermatitis mit lymphohistiozytären Zellen, eosinophilen Granulozyten, Mastzellen und neutrophilen Granulozyten. Diese Muster stimmt mit einer Hypersensibilitätsreaktion vom Spättyp auf Pilzantigene überein. Spezialfärbungen erbrachten keinen Hinweis auf eine Hypersensibilität der kutanen basophilen Granulozyten bei diesen Reaktionen. Unsere Befunde deuten en, daß natürlich auftretende Dermatomykosen bei Katzen von der Entwicklung sowohl humoraler als auch zellulärer Immunantworten auf diese Organismen begleitet werden. Intradermale Hauttests mit Dermatophytenantigenen sind ein nützliches Verfahren um die Immunantwort von Katzen auf Dermatomykosen zu untersuchen. Resumen Para estudiar la respuesta inmune de los gatos hacia la infección por dermatofítos, diez gatos sanos y cinco gatos previamente infectados con Microsporum canis fueron inyectados intradérmicamente con extractos de glicoproteina de dermatofito. En los lugares de inyección se observaron las reacciones inmediatas y posteriores y se, realizaron biopsias a los 24 y 48 horas después de la inyección. Tuvieron reacciones positivas inmediatas hacia el antígeno M. canis los cinco gatos prevaimente infectados y uno de los diez “gatos control”. Cuatro de los gatos previamente infectados y uno de los diez “gatos control” tambien reaccionaron ante el antígeno Tríchophyton rubrum, sugeriendo reactividad cruzada, antigénica entre estas especies fúngicas. A los 24 horas después de la inyección, cuatro des los cinco gatos previamente infectados presentaron reacciones posteriores hacia el antígeno M. canis en los lugares de la inyección; los lugares equivalentes en todos los animales control fueron negativos. La aparición histológica de las reacciones posteriores consistía en una dermatitis perivascular superficial, con celulas linfohistiocíticas, eosinofilos, mastocitos y neutrofilos. Este patrón es consistente con una hipersensibilidad de dipo retrasado en respuesta a antígenos fúngicos. Manchas especiales revelaron la no evidencia de hipersensibilidad de basófilos cutaneous en estas reacciones. Nuestros datos sugieron que la dermatofitosis felina producida de forma natural va acompañada del desarrollo de respuestas inmunes humorales y celulares hacía el organismo. La prueba intradérmica con antígenos dermatofítos es útil para investigar la respuestra inmunológica de los gatos hacia dermatofitosis.     

Drug Efficacy of Terbinafine Hydrochloride (Lamisil®) During Oral Treatment of Cats,Experimentally Infected with Microsporum canis

Cats represent a primary source of Microsporum canis infections in humans. Terbinafine hydrochloride (Lamisil^®) is commonly used in the treatment of microsporosis in humans as its fungicidal action permits short periods of treatment. The aim of the present study was to estimate the efficacy of the drug in cats. Nine cats were experimentally infected with M. canis and treated with terbinafine hydrochloride at a dose of 10–20 mg/kg (once daily, SID; low‐dose group, LDG). Another nine cats were similarly infected and treated with 30–40 mg/kg SID (high‐dose group, HDG) and a further nine cats were also infected and left untreated (control group, CG). The general condition of the cats was observed daily and their clinical symptoms evaluated weekly. The cats recovery was monitored using the Wood's lamp illumination test and microscopic and fungal culture examinations. The general condition of the cats during the study was good. The cure rates of the LDG were not significantly different from the CG at any period during the treatment. However, the HDG cure rates differed significantly from the other two groups. After 109 days of treatment, when all nine cats of the HDG were healed, seven cats of the LDG and all the cats in the CG were still M. canis‐positive. This study shows that dosages of 10–20 mg/kg SID of terbinafine hydrochloride are not sufficient to terminate an experimental M. canis infection in cats within an acceptable period of time. Terbinafine hydrochloride can be used to treat dermatophytosis in cats, but a higher dosage, 30–40 mg/kg SID, should be used to achieve a cure.     

Dermatophytes isolated from domestic and feral animals

In work over a period of 8 years, dermatophytes were recovered from 12 animal species in the North Island of New Zealand. A total of 552 dermatophytes were isolated and belonged to the Microsporum (6 species) and Trichophyton (6 species) genera.

Some unusual isolations are reported: Microsporum canis and Trichophyton mentagrophytes var. mentagrophytes were recovered from calves; Microsporum distortum from a dog, which was the first known isolation from an animal in the North Island; four horses, from the same stable, yielded Microsporum equinum which has not previously been recorded in this country; Trichophyton erinacei was recovered from lesions on a cat which is the first report of the dermatophyte from an animal other than the dog, hedgehog or man; Trichophyton equinum var. autotrophicum was a rare isolation from a dog.

Species affinity was demonstrated with the dermatophytes M. canis; M. nanum; M. equinum; T. equinum; and T. verrucosum. These zoophilic species appear to be passed between individuals of the same species, with occasional infection in man and other animal species. T. mentagrophytes var. mentagrophytes had a wider distribution, being isolated from dogs, cats, guinea-pigs and rats. The infection in rats was subclinical.

The geophilic species M. cookei, T. ajelloiand and T. terrestre were recorded but not regarded as being pathogenic. M. gypseum was significant in cases involving dogs, horses and a cat, as arthrospores were seen invading the affected hairs.     

Treatment of Microsporum canis ringworm in a cat colony

Abstract— The treatment and control measures used during an outbreak of feline ringworm caused by Microsporum canis are described. During the outbreak the presence of the dermatophyte on the coats of clinically normal cats was demonstrated by the use of Mackenzie's brush technique. This method of examination is suggested as being the most effective and rapid means of ensuring that dermatophyte infection is not introduced into a colony by new cats. Résumé— L'auteur décrit le traitement et les mesures antiparasitaires appliqués à l'occasion d'une épizootie de teigne féline, provoquée par Microsporum canis. La préence du dermatophyte sur le poil de chats cliniquement normaux a été décelée par la technique de la brosse Mackenzie, méthode particulièrement rapide et efficace, propre à éviter la contamination d'une colonie de chats par des dermatophytes importés de l'extérieur. Zusammenfassung— Die Behandlung und die Kontrollmassnahmen während eines Ausbruchs von durch Microsporum caniJ verursachter Katzenringwurmkrankheit werden beschrieben. Wahrend der Krankheit wde das Vorhandensein der Dermatophyten in den Pelzen klinisch normaler Katzen durch Anwendung der Mackenzieschen Pinselmethode demonstriert. Diese Untersuchungs methode wird ah wirksamste und schnellste vorgeschlagen, die gewährleistet, dass die Dermatophyten nicht durch neue Katzen in eine Kolonie eingeschleppt werden.     

Assessment of immunogenicity and protective efficacy of Microsporum canis secreted components coupled to monophosphoryl lipid-A adjuvant in a vaccine study using guinea pigs

Microsporum canis is the most common dermatophyte in pets and is of zoonotic importance but currently there is no effective vaccine available to prevent dermatophytosis. The aim of this work was to assess the immunogenicity and protective efficacy of secreted components (SC) from M. canis adjuvanted with the monophosphoryl lipid-A (MPLA), in a vaccine study using the guinea pig as an experimental model. Animals were vaccinated with either the SC adjuvanted with the MPLA, the MPLA adjuvant alone or PBS three times at two-week intervals, until 42 days prior to M. canis infection. A blind evaluation of dermatophytosis symptoms development and fungal persistence in skin was monitored weekly. The antibody response towards the SC and the levels of Interferon (IFN)γ and Interleukin-4 expressed in peripheral blood mononuclear cells were assessed along or at the end of the study period respectively. The animals that received MPLA had a significantly lower clinical score than those inoculated with PBS. However, no significant difference was observed between the guinea pigs vaccinated with the SC adjuvanted with the MPLA and those having received MPLA alone. The results also showed that vaccination induced a strong antibody response towards the SC and an increase in IFNγ mRNA level. Our results show that the MPLA adjuvant used in this vaccine study can induce per se a partial protection against a M. canis infection. Although they induce a delayed-type hypersensitivity reaction in guinea pigs, the SC do not confer a protection under the present experimental conditions.     

Isolation of Dermatophytes from the Haircoats of Stray Cats from Selected Animal Shelters in two Different Geographic Regions in the United States

Résumé— Le but de cette étude était de déterminer la prévalence des isolements de Microsporum canisà partir de chats apparemment sains dans des refuges de deux régions de climats différents aux Etats-Unis. Deux cents chats (chats sans signes apparent de maladie) provenant d'une région froide et séche et d'une région chaude et humide ont étéétudiés et la technique de la brosse à dents utilisée pour la mise en culture et la rechehe de Dermatophytes. Cinq espèces de Dermatophytes communément associées aux dermatophyties humaines et animales ont été isolées: M. canis (n= 8), Trichophyton verrucosum (n= 5), T. mentagrophytes (n - 1), T. rubrum (n= 16), et Epidermophyton spp. (n= 5). Microsporum canis n'a été isolé que chez les chats vivant dans le Sud et principalement sur des chatons. Trichophyton rubrum a surtout été isolé chez les chats du nord. La prévalence de M. canis chez les chats sains était dans cet étude de 4 pourcent et la prévalence totale de tous les pathogénes de 17,5 pour cent. [Isolation of dermatophytes from the haircoats of stray cats from selected animal shelters in two different geographic regions of the United States. (Isolement de dermatophytes du pelage de chats errants pris dans des refuges de deux regions différentes des Etats-Unis.) Zusammenfassung— Ziel der Studie war, die Prävalenz von Microsporum canis-Isolaten von scheinbar gesunden Katzen in Tierheimen in zwei klimatisch unterschiedlichen Regionen der Vereinigten Staaten festzustellen. Zwei hundert Katzen (Katzen ohne makroskopische oder offenscichtliche physische Anzeichen von Krankheit) aus Tierheimen in einer relativ kalt-trockenen geographischen Region und einer feucht-war-men Region der Vereinigten Staaten wurden untersucht und aus dem Haarkleid mittels Zahnbürstentechnik Pilzkulturen angelegt. Es wurden fünf Arten von Dermatophyten identifiziert, die häufig mit menschlichen oder tierischen Hautpilzerkrankungen verbunden sind: M. cams (n= 8), Trichophyton verruscosum (n= 5), T. mentagrophytes (n= 1), T. rubrum (n= 16) und Epidermophyton spp. (n= 5). Microsporum canis wurde nur von Katzen aus dem Süden isoliert, vorwiegend von Katzenwelpen. Trichophyton rubrum wurde häufiger von Katzen aus dem Norden isoliert. Die Prävalenz von M. canis bei scheinbar gesunden Katzen in dieser Studie betrug 4 Prozent, die Gesamtprävalenz aller pathogenen Keime betrug 17,5 Prozent. [Isolation of dermatophytes from the haircoats of stray cats from selected animals shelters in two different geographic regions in the United States (Isolierung von Dermatophyten aus dem Fell von streunenden Katzen aus ausgewahlten Tierheimen in awei verschiedenen geographischen Regionen der Vereinigten Staaten.) Abstract— The purpose of this study was to determine the prevalence of Microsporum canis isolation from apparently healthy cats in shelters in two different climatic regions of the United States. Two hundred cats (cats without gross or obvious physical illness) from animal shelters in a relatively cold dry geographic region and a warm humid region of the United States were examined and their haircoat cultured for fungi using the toothbrush technique. Five species of dermatophytes commonly associated with either animal or human dermatophytosis were identified: M. canis (n= 8), Trichophyton verruscosum (n= 5), T. mentagrophytes (n= 1), T. rubrum (n= 16), and Epidermophyton spp. (n = 5). M. canis was isolated only from cats in the south, predominantly from kittens. T. rubrum was isolated more commonly from cats from the north. The prevalence of M. canis in apparently healthy cats in this study was 4 per cent and the overall prevalence of all pathogens was 17.5 per cent.     

Suppurative spinal meningomyelitis in a dog with intra-neutrophilic cerebrospinal fluid cells Ehrlichia canis morulae

A 2-year-old castrated male Creole Shepherd mixed dog was presented for non-ambulatory paraparesis of the pelvic limbs. The magnetic resonance imaging and cerebrospinal fluid (CSF) analysis were consistent with meningomyelitis. Positive serology for Ehrlichia canis/Ehrlichia ewingii suggested exposure to a pathogen; qPCR on the serum and the CSF confirmed active infection. Ehrlichial morulae were observed within CSF and peripheral blood polymorphonuclear neutrophils; a species-specific PCR confirmed E. canis infection. This is an interesting report of E. canis infection in a dog with morulae observed in neutrophils both in the peripheral blood and CSF.     

Evaluation of Fungicidal Efficacy of Benzalkonium Chloride (Steramina G u.v.) and Virkon-S against Microsporum canis for Environmental Disinfection

The aim of this study is to evaluate the antifungal efficacy of Steramina G u.v. (10% solution of alkyldimetylbenzylammonium chloride; Formenti Grünenthal) and Virkon-S (multipurpose system; Antec International) against Microsporum canis-infected hairs and spores. Samples were collected from a random sample of household cats and from subjects from catteries. Seventy M. canis-positive hairbrushes containing furs, keratin scales and other organic material were treated with each of the two disinfectants, using concentrations recommended by the manufacturer's instructions (2% and 1% for Steramina G u.v. and Virkon-S, respectively). Each brush remained in contact with the antifungal solution for 10 min. After this period, the brushes were air-dried, then seeded into mycobiotic agar, and incubated for up to 21 days at 28°C. The disinfectants were considered effective if dermatophytes failed to grow. Steramina G u.v. was effective in 97.14% of samples and Virkon-S in 87.14%. The antifungal activity of Steramina G u.v. against M. canis was significantly higher (p < 0.05) than that of Virkon-S.     

Periparturient immunosuppression in the bitch and its influence on infection with Toxocara canis

Pregnancy and lactation in dogs induced a marked suppression of their immunological responsiveness as judged by in vitro phytomitogen- and Toxocara canis antigen-induced lymphocyte transformation. In addition, the eosinophilia otherwise seen following infection with T. canis was suppressed during the periparturient period. Coincidental with this periparturient immunosuppression was the occurrence of heavy infections with T. canis in the intestine of lactating bitches and the establishment of such infections perhaps may be related to the suppressed immune reactivity observed. These infections in the bitch appeared to be acquired primarily through the ingestion of immature stages of T. canis passed in the faeces of their puppies. The results demonstrate that the lactating bitch can be a major source of contamination of the environment with eggs of T. canis.     

Hepatozoon canis: The prevalence of antibodies and gametocytes in dogs in Israel

A survey for the prevalence of antibodies to Hepatozoon canis and for intraneutrophilic H. canis gametocytes in the peripheral blood neutrophils of dogs in Israel showed that 33.1% were seropositive, while only 1% of the dogs sampled had detectable parasites in their blood smears. Exposure to H. canis is widespread but it appears that most infected dogs undergo a subclinical infection and only a small proportion develop clinical disease.Abbreviations IFAT indirect fluorescent antibody test     

Validation of an ELISA method for the serological diagnosis of canine brucellosis due to Brucella canis

In the present study, the validation of an enzyme-linked immunosorbent assay (ELISA) for serodiagnosis of canine brucellosis is described. Two different antigenic extracts, obtained by heat or ultrasonic homogenization of microbial antigens from a wild isolate of Brucella canis bacteria, were compared by ELISA and Western blot (WB). A total of 145 canine sera were used to define sensitivity, specificity and accuracy of the ELISA as follows: (1) sera from 34 animals with natural B. canis infection, confirmed by blood culture and PCR, as well as 51 sera samples from healthy dogs with negative results by the agar–gel immunodiffusion (AGID) test for canine brucellosis, were used as the control panel for B. canis infection; and (2) to scrutinize the possibility of cross reactions with other common dog infections in the same geographical area in Brazil, 60 sera samples from dogs harboring known infections by Leptospira sp., Ehrlichia canis, canine distemper virus (CDV), Neospora caninum, Babesia canis and Leishmania chagasi (10 in each group) were included in the study. The ELISA using heat soluble bacterial extract (HE-antigen) as antigen showed the best values of sensitivity (91.18%), specificity (100%) and accuracy (96.47%). In the WB analyses, the HE-antigen showed no cross-reactivity with sera from dogs with different infections, while the B. canis sonicate had various protein bands identified by those sera. The performance of the ELISA standardized with the heat soluble B. canis antigen indicates that this assay can be used as a reliable and practical method to confirm infection by this microorganism, as well as a tool for seroepidemiological studies.     

Prenatal and transmammary infections of Toxocara canis in dogs: effect of benzimidazole-carbamate anthelmintics on various developmental stages of the parasite

Fenbendazole and albendazole, given at a dose rate of 150 mg/kg for 3 days, produced a 90 per cent reduction in the numbers of second stage larvae of Toxocara canis present in the tissues of dogs although no reduction in the number of larvae found in the brains of infected dogs occurred with this treatment. The results suggest that a course of 3 day therapy with these anthelmintics should prevent prenatal infections in puppies. However, if infection is acquired by bitches during late pregnancy or early lactation, the transmammary route of infection becomes important. Therefore, anthelmintic treatment of the bitch prior to pregnancy will not prevent transmission of infection to her puppies should the bitch acquire a new infection of T. canis during pregnancy or early lactation. Alternatively, infection with T. canis can be controlled through the treatment of neonatal puppies for migrating larvae of T. canis. Treatment of newborn puppies with fenbendazole, albendazole or oxfendazole at a dose of 100 mg/kg for 2–3 days produced a 91–99 per cent reduction in the number of adult parasites found. In addition, a single dose of fenbendazole, given at a dose rate of 40 mg/kg, eliminated 93–96 per cent of adult T. canis from the intestines of 4–5-week-old puppies. These latter treatments would need to be repeated to eliminate completely the infection from puppies.     

Human Brucella canis Infection and Subsequent Laboratory Exposures Associated with a Puppy,New York City, 2012

Human Brucella canis infection incidence is unknown. Most identified cases are associated with pet dogs. Laboratory‐acquired infections can occur following contact with Brucella spp. We identified a paediatric B. canis case, the source and other exposed persons. A 3‐year‐old New York City child with fever and dyspnoea was hospitalized for 48 h for bronchiolitis. After her admission, blood culture grew B. canis, she was prescribed anti‐microbials and recovered. B. canis was also isolated from blood of the child's pet dog; these isolates were genetically similar. The dog originated from an Iowa breeding facility which was quarantined after identification of the dog's infection. Additionally, 31 laboratory workers were exposed and subsequently monitored for symptoms; 15 completed post‐exposure prophylaxis. To our knowledge, this is the first report strongly suggesting B. canis zoonotic transmission to a child in the United States, and highlights the need for coordinated control policies to minimize human illness.     

Efficacy of eight commercial formulations of lime sulphur on in vitro growth inhibition of Microsporum canis

Lime sulphur is a common topical treatment for dermatophytosis in animals. Until recently, a single veterinary lime sulphur formulation was available. The purpose of this study was to compare the efficacy of eight lime sulphur products for in vitro growth inhibition of Microsporum canis using the isolated infected spore model. Infective M. canis spores were isolated from hairs collected from untreated cats. Hairs were macerated in Triton‐X solution and isolated according to a previously published protocol. Equal volumes of spore suspension and lime sulphur solutions were incubated for 5 min and plated onto modified BBL? Mycosel? agar (Becton, Dickinson and Company; Sparks, MD, USA) plates. Five plates were inoculated for each sample solution. Distilled water and bleach were used as controls. Colony forming units were counted daily for 21 days; positive control plates contained >300 colony forming units/plate. Seven of the products were supplied as concentrates and they were tested at the manufacturer’s recommended dilution, twice label concentration and half label concentration. A prediluted product SulfaDip^® (Trask Research, Inc.; Daluca, GA, USA) was tested at the label and half label concentration. All veterinary products formed recommended treatment dilutions of 3% sulphurated lime solution except one (LymDyp^®, IVX Animal Health Inc.; St Joseph, MO, USA), which formed a 2.4% sulphurated lime solution. Results of the study showed complete growth inhibition of M. canis spores by all products at all dilutions tested. These results indicate that all tested lime sulphur‐containing products were equivalent. Field studies are needed to test product equivalency in vivo.