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1.
In western immunoblotting studies of canine sera using Malassezia pachydermatis extracts, the reported patterns of immunoreactivity vary between different laboratories. Because the duration of culture influences the antigenic composition of lipid-dependent Malassezia spp. when probed with human sera, we investigated whether the in vitro growth phase of M. pachydermatis influences immunoreactivity using canine sera. Extracts of M. pachydermatis CBS 1879 grown in Sabouraud's liquid medium at 37 degrees C for 2, 4, 6, 8 and 10 days were prepared by mechanical disruption, centrifugation, dialysis and lyophilization. Yeast growth phase was assessed by sequential colony counts and optical density measurements. Patterns of IgG immunoreactivity in high (n = 3) and low (n = 3) titre sera were compared using extracts prepared at each time point by sodium dodecyl sulphate polyacrylamide gel electrophoresis and western immunoblotting. Protein bands of 62 and 49 kDa were recognized by all sera, and 98 and 68 kDa bands were recognized by five sera. Proteins of 188, 66, 58, 57, 38, 28 and 17 kDa were only recognized by high titre sera. All high titre sera used recognized more bands in exponential phase (d2) extracts when compared with decline phase (d8-d10) extracts, and two of these sera showed most bands in stationary phase (d4-d6) extracts. Bands of 62 and 57 kDa were primarily detected in exponential and early stationary phase extracts. There is variation in antigenic expression in different growth phases of M. pachydermatis, which might explain discrepancies between previous laboratory studies of canine immunity to this yeast.  相似文献   

2.
Western immunoblotting studies of canine sera using Malassezia pachydermatis extracts have shown that infected dogs commonly have antibodies that recognize multiple antigens. However, reported patterns of immunoreactivity vary between different laboratories. Since culture duration influences the antigenic composition of lipid‐dependent Malassezia when probed with human sera, we investigated whether the in vitro growth phase of M. pachydermatis influences immunoreactivity to canine sera. Extracts of M. pachydermatis CBS1879 grown in Sabouraud's liquid medium at 37°C for 2, 4, 6, 8 and 10 days were prepared by mechanical disruption, centrifugation, dialysis and lyophilization. Yeast growth phase was assessed by sequential colony counts and optical density measurements. Patterns of IgG immunoreactivity in high (n = 3) and low (n = 3) titre sera were compared using extracts prepared at each time point by SDS‐PAGE and western immunoblotting. Protein bands of 62 and 49 kDa were recognized by all sera, and five sera recognized 98 and 68 kDa bands. Proteins of 188, 66, 58, 57, 38, 28 and 17 kDa were only recognized by high titre sera. All high titre sera recognized more bands in exponential phase (day 2) extracts when compared with decline phase (days 8–10) extracts, and two of these sera showed most bands in stationary phase (days 4–6) extracts. Bands of 62 and 57 kDa were primarily detected in exponential and early stationary phase extracts. Antigenic variation in extracts of M. pachydermatis prepared during different growth phases might explain discrepancies between previous laboratory studies of immunity to this yeast. Funding: Government of Malaysia.  相似文献   

3.
Abstract We have previously shown that both atopic and normal dogs generate an IgG response to antigens of Malassezia pachydermatis . The aim of this study was to compare IgE responses to separated proteins of M. pachydermatis in 28 atopic dogs with Malassezia dermatitis and 22 clinically normal dogs using Western immunoblotting. Six different detection systems were evaluated in order to assess sensitivity and eliminate nonspecific binding and cross-reactivity. The protocol yielding the best results utilized a monoclonal mouse antidog IgE, an alkaline phosphatase conjugated goat antimouse IgG which had been passed through a canine IgG column 3 times, a chemiluminescent substrate and a digital imaging system. Proteins of 45, 52, 56 and 63 kDa were recognized by more than 50% of the atopic dog sera and thus represented major allergens. Only a minority of normal dogs showed faint IgE binding to these proteins. The results indicate that the majority of atopic dogs with Malassezia dermatitis have a greater IgE response than normal dogs, suggesting an IgE-mediated immune response may be clinically important in the pathogenesis of the disease.  相似文献   

4.
IgG immunoreactivity to Malassezia pachydermatis was compared in atopic and non-atopic dogs. Malassezia pachydermatis proteins with a molecular weight of 98 kDa were recognized at a significantly higher frequency in the sera of atopic dogs. Most of the atopic dogs with Malassezia dermatitis had a greater IgG response than did normal dogs.  相似文献   

5.
The aim of this study was to determine the in vitro antifungal activity of several antifungal drugs (posaconazole, nystatin, miconazole and clotrimazole) against Malassezia pachydermatis with microdilution and agar dilution techniques. Malassezia pachydermatis isolates were obtained from the skin and ears of dogs. Tests on solid media were performed using 25-well Petri dishes (2 mL/well containing Sabouraud's dextrose agar and diluted antifungal drug) inoculated with 5 μL suspensions of M. pachydermatis . Microtitre broth dilution used 96-well microtitre plates containing Sabourauds dextrose broth and appropriate dilutions of antifungal drugs, inoculated with 10 μL standard suspensions of M. pachydermatis . Plates were inoculated in duplicate and incubated at 30°C for 5 days and growth assessed. The four antifungal drugs were tested in 10 dilutions (4.0-0.007 μg/mL for posaconazole, and 32--0.06 μg/mL for clotrimazole, miconazole and nystatin). Results obtained for 83 strains of M. pachydermatis and a control reference strain (CBS 1879) exhibited the same pattern. Results of the MIC between microtitre and agar methodologies showed no significant differences (≤ 2-fold) across all drugs. For both solid and liquid methods, posaconazole was the most effective antifungal drug of the four tested with MIC90 of 1–2 μg/mL for posaconazole, 16–32 μg/mL for clotrimazole, and ≥ 32 μg/mL for miconazole and nystatin.
Funding: Schering-Plough.  相似文献   

6.
OBJECTIVE: To investigate the direct interaction between canine keratinocytes and live Malassezia pachydermatis and thereby determine the role of these organisms in the pathogenesis of epidermal hyperplasia associated with Malassezia dermatitis in dogs. SAMPLE POPULATION: Primary canine keratinocyte cultures established from skin samples obtained from clinically normal dogs. PROCEDURE: The proliferative response of keratinocytes co-cultured with Malassezia organisms for 1, 2, or 3 days was assessed by use of direct manual counting (to determine the number of keratinocytes in both the monolayer and the medium) and immunohistochemical staining techniques involving antibodies against proliferating cell nuclear antigen (PCNA) and another cellular proliferation marker, Ki-67. The potential cytotoxic effect of Malassezia organisms was investigated by use of an apoptosis detection kit to label keratinocytes co-cultured with M. pachydermatis that underwent apoptosis. RESULTS: No stimulatory effect of Malassezia organisms on canine keratinocyte proliferation was detected via cell counting and immunohistochemical techniques. However, there was a significant increase in dead keratinocytes in the medium with increasing numbers of Malassezia organisms in the co-culture. More apoptotic cells were observed in keratinocyte monolayers co-cultured with high numbers of M. pachydermatis than there were in monolayers cultured without Malassezia organisms, and the number increased after prolonged incubation. CONCLUSIONS AND CLINICAL RELEVANCE: M. pachydermatis did not stimulate canine keratinocyte proliferation in vitro. The results suggested that the epidermal hyperplasia observed in dogs with Malassezia dermatitis is unlikely to be caused by a direct effect of the organism on the keratinocyte cell cycle, but is likely to involve other mechanisms.  相似文献   

7.
Serum immunoglobulin G (IgG) responses of healthy dogs and dogs with Malasseziapachydermatis dermatitis were compared by Western immunoblotting. M pachydermatis CBS 1879 was disrupted mechanically and its proteins were separated and blotted on to nitrocellulose membranes before being incubated with sera from eight healthy beagles, eight Irish setters with gluten-sensitive enteropathy, 15 healthy basset hounds, and 30 dogs with Mpachydermatis-associated dermatitis, 20 of which were basset hounds. The mean (se) numbers of bands of immunoreactivity observed in the seborrhoeic basset hounds (10.7 [0.4]) and affected mixed-breed dogs (9.4 [0.9]) were significantly greater than in the beagles (3-0 [1.0]), Irish setters (5.5 [1.1]) and healthy basset hounds (5.6 [0.7]). The number of bands identified was correlated (r(s) = 0.76, P < 0.001) with the anti-M pachydermatis IgG values measured by ELISA in a previous study. Most of the dogs were immunoreactive towards the 132, 66 and 50 to 54 kDa proteins and the affected dogs were also usually reactive towards the 219, 110, 71 and 42 kDa proteins.  相似文献   

8.
Abstract Epidermal hyperplasia is one of the major histopathological features seen in dogs with Malassezia dermatitis. The aim of this study was to investigate the effects of extracts and culture supernatants from Malassezia pachydermatis on the proliferation of canine keratinocytes. Keratinocyte cultures were established from normal dog skin, and cell monolayers were co-cultured with Malassezia extracts (prepared either with or without protease inhibitors) and supernatants derived from organisms grown in liquid culture. The proliferation of keratinocytes was measured using a colourimetric assay. Neither the culture supernatants nor the Malassezia extracts had significant effects on the proliferation rate of canine keratinocytes, regardless of whether protease inhibitors were present or not. The results indicate that the epidermal hyperplasia seen in Malassezia dermatitis is unlikely to be caused directly by secretion of products from the organism.  相似文献   

9.
Malassezia pachydermatis is considered to be a contributing factor to canine atopic dermatitis (AD). The purpose of this study was to investigate the humoral response to a commercially produced M. pachydermatis extract. Fifteen atopic dogs with Malassezia overgrowth on the skin (MD), 16 atopic dogs without MD, three atopic dogs with overgrowth of Malassezia in the ears only (MO), and 12 normal dogs were intradermally tested with M. pachydermatis extract at 50, 100, 250, 500, 1000, 2000 and 4000 PNU mL(-1). All dogs were evaluated cytologically by cutaneous tape strip and bilateral ear exudate sampling to determine presence of MD or MO. Each had serum evaluated for anti-Malassezia IgE using three Malassezia extracts with an ELISA assay. The irritant threshold concentration at which healthy nonatopic dogs ceased to react was 1000 PNU mL(-1). There was a significant difference in intradermal test reactivity between the atopic groups. At this dilution, 93% (14/15) of the atopic MD group, 31% (5/16) of the atopic group without MD or MO, and 100% (3/3) of the atopic MO only group reacted. There were no significant differences in the serum IgE levels as measured by the Greer ELISA assay, between any groups using any of the three extracts. These results support that Greer's M. pachydermatis extract is useful for intradermal testing of dogs with an allergic phenotype, and that atopics with MD are more likely to have a type-1 Malassezia hypersensitivity than those without. The ELISA assay may require further development in order to be useful for the diagnosis of Malassezia hypersensitivity.  相似文献   

10.
Eight beagles were experimentally inoculated intraotally with Malassezia pachydermatis to induce acute otitis externa. Three or 4 days after the inoculation, the animals showed the symptoms of otitis externa. All ear canals were erythematous and the dogs were shaking their heads. A large number of M. pachydermatis was noticed in exudate taken from every ear canal. Clinical signs of otitis externa were reduced after treatment with 0.1 ml (per canal) of 1% pimaricin suspension twice a day for 3 days. The amount of exudate decreased gradually and 12 of the 16 ear swabs examined, thereafter, were found to be negative for M. pachydermatis within 10 days. No side effects were observed in all the treated cases. These results suggested that M. pachydermatis could induce the canine otitis externa, and that pimaricin is effective agent for M. pachydermatis infection in ear canals.  相似文献   

11.
Abstract Suspensions of Malassezia pachydermatis adhered to canine corneocytes attached io adhesive tape in a dose (P < 0.001) and time-dependent (P < 0.01) manner; adherence was maximal after 2 h. M. pachydermatis cells were approximately 10 times more adherent than Saccharomyces cerevisiae (P < 0.001) cells after 2 h incubation. The adherence of formalin-treated and frozen-thawed M. pachydermatis cells was comparable with untreated controls. Stationary-phase cells adhered better (P < 0.05) than exponential-phase cells. Pretreatment of the yeasts, or corneocytes, with 0.1% trypsin for 30 min reduced (P < 0.01) the adherence of four, and two, out of five strains, respectively, whereas incubation with 300 mM solutions of D(+) mannose, sucrose and N-acetyl D-glucosamine had no consistent effect. These results suggest that trypsin-sensitive proteins or glycoproteins on the yeast cell wall, and on the corneocyte surface, play an important role in the adherence of M. pachydermatis to canine corneocytes in vitro, whereas a role for carbohydrate receptors was not demonstrated. Résumé— Des suspensions de Malassezia pachydermatis adhérant à des cornéocytes des chiens sont attachées à des rubans adhésifs de façon significative en function du nombre (P < 0,001) et de la durée (P < 0,01). L'adhérence est maximale après 2 heures. Les levures du genre Malassezia pachydermatis sont approximativement dix fois plus adhérentes que les levures du genre Saccharomyces cerevisiae (P < 0,001) après une incubation de 2 heures. L'adhérence des Malassezia pachydermatis traitées par le formol et congelées - décongelées, est comparable à celle des témoins. Les levures qui ne sont pas en phase de croissance adhérent mieux (P < 0,05) que celles qui le sont. Le traitement préalable des levures ou des cornéocytes avec une solution de trypsine pendant 30 minutes réduit (P < 0,01) l'adhérence tandis que l'incubation avec des solutions à 300 mM de D + mannose, sucrose et de N acétyl D glucosamine n'a pas d'effets. Ces résultats suggèrent que des protéines sensibles à la trypsine ou des glycoprotéines sur la paroi des levures et à la surface des cornéocytes jouent un rôle important in vitro dans l'adhérence des Malassezia pachydermatis aux cornéocytes du chien, alors que le rôle des récepteurs glucidiques n'a pas été démontré. [Bond, R., Lloyd, D. H. Factors affecting the adherence of Malassezia pachydermatis to canine cornéocytes in vitro (Facteurs influençant l'adhérence de Malassezia pachydermatis aux cornéocytes du chien in vitro). Veterinary Dermatology 1996; 7 : 49–56.] Resumen Las suspensiones de Malassezia pachydermatis se adherian a cinta adhesiva de forma dependiente de la dosis (P < 0.001) y del tiempo (P < 0.01); su adherencia fue máxima a las 2 h. Las células de M. pachydermatis fueron aproximadamente 10 veces más adherentes que las de Saccharomyces cerevisiae (P < 0.001) a las 2 h de incubación. La adherencia de células de M. pachydermatis tratadas con formalina y congeladas-descongeladas fue comparable con los controles no tratados. Las células en estadio estacionario se adherian mejor (P < 0.05) que las de fase exponencial. El tratamiento previo de las levaduras o los corneocitos con 0.1% de tripsina durante 30 min redujo (P < 0.01) la adherencia de cuatro, y dos, de cinco cepas, respectivamente, mientras que su incubación con soluciones 300 mM de D(+) manosa, sucrosa y N-acetil D-glucosamina no tuvieron un efecto constante. Estos resultados sugieren que proteinas o glieoproteinas sensibles a la tripsina en la pared de la levadura, y en la superficie del corneocito, juegan un papel importante en la adherencia de M. pachydermatis a los corneocitos caninos in vitro, mientras que no se pudo demostrar un papel por parte de los receptores de carbohidratos. [Bond, R., Lloyd, D. H. Factors affecting the adherence of Malassezia pachydermatis to canine corneocytes in vitro (Facto res que afectan la adherencia de Malassezia pachydermatis a los corneocitos caninos in vitro). Veterinary Dermatology 1996; 7 : 49–56.] Zusammenfassung— Suspensionen von Malassezia pachydermatis zeigten eine Adhärenz an kanine Korneozyten, die an einem Klebeband befestigt waren, in dosisabhängiger (P < 0,001) und zeitabhängiger Weise (P < 0,01). Die Adhärenz erreichte ein maximum nach 2 Stunden. M. pachydermatis-Zellen waren ungefähr 10 mal stärker adhärent als Saccharomyces cervisiae-Zellen nach Zstündiger Inkubation (P < 0,001). Die Adhärenz von formalinbehandelten und gefrorenen/aufgetauten M. pachydermatis-Zellen war vergleichbar mit unbehandelten Kontrollzellen. Zellen der stationären Phase waren besser adhärent (P < 0,05) als Zellen der exponentiellen Phase. Eine Vorbehandlung der Hefen oder Korneozyten mit 0,1% igem Trypsin über 30 Minuten reduzierte die Adhärenz (P < 0,01) von 4 bzw. 2 aus 5 Linien, während eine Inkubation mit 300 mm Lösungen von D(+)Mannose, Sucrose und N-Acetyl-D-Glukosaminen keinen entsprechenden Effekt hatte. Diese Ergebnisse legen nahe, daß Trypsin-sensible Proteine oder Glykoproteine an der Zellwand der Hefe und auf der Korneozytenoberfläche eine wichtige Rolle für die Adhärenz von M. pachydermatis an kanine Korneozyten in-vitro spielen, während eine Bedeutung für Kohlenhydrat-Rezeptoren nicht demonstriert werden konnte. [Bond, R., Lloyd, D. H. Factors affecting the adherence of Malassezia pachydermatis to canine corneocytes in vitro (Einflußfaktoren auf die Adhärenz von Malassezia pachydermatis an kanine Korneozyten in vitro). Veterinary Dermatology 1996; 7 : 49–56.]  相似文献   

12.
To investigate the role of Malassezia pachydermatis as a pathogenic agent in canine otitis, a comparative analysis of isolates from normal and diseased external ear canals in dogs was undertaken. Specimens were collected from the ears of dogs with unilateral or bilateral otitis and from healthy dogs. Mycological analysis was by direct microscopy and fungal culture on Sabouraud's dextrose agar and Dixon's agar. Of the otitis specimens, 63.7% showed typical Malassezia cells on cytological examination. In samples taken from the healthy ears of dogs with unilateral otitis, only 21.43% (P<0.05) showed evidence of Malassezia. M. pachydermatis was identified cytologically and culturally in 57.53% (P<0.05), 14.29% and 30.0% of samples from the ears of dogs with otitis, from the healthy ears of dogs with unilateral otitis and from the ears of healthy dogs with no otitis. In the group with otitis associated with M. pachydermatis, the poodle was the most common breed (39.29%; P<0.05), whereas in the group without otitis, the German Shepherd breed was prominent (although this observation was not statistically significant). In both groups, the majority of dogs with M. pachydermatis were aged between 1 and 3 years (P<0.05). The higher incidence of M. pachydermatis isolated from the ears of dogs with otitis externa suggests a putative pathogenic role of this yeast in this condition.  相似文献   

13.
The chronological development of the serum IgE and IgG response to microfilaria, third and fourth stage larvae, and male and female adult Dirofilaria immitis was examined by enzyme-linked immunosorbent assay (ELISA) and enzyme-linked immunoelectrotransfer blot (EITB). Dirofilaria immitis-specific IgE and IgG levels peaked 16-18 weeks post-infection after increasing in response to the fourth larval molt. Specific IgG levels plateaued after patency, while IgE continued to decline. The use of ammonium sulfate cut sera showed there was no quenching or blocking of IgE binding by IgG in the ELISA and EITB methods used in this study. IgE-specific EITB showed 30-49 bands for the five respective extracts that were identified by M(r) or relative mobility. Eighty-five to 100 bands were visualized by IgG-specific EITB for the same five extracts. The isotype-specific ELISA and EITB were shown to be closely related by significant correlations (P < 0.0001) between S/N ratios and the number of bands found on blots. The isotype-specific EITB bands non-specifically recognized were greater in size than 21 kDa for IgG and 45 kDa for IgE. Recognition of bands changed over time with some bands being recognized only by prepatent sera. Ten antigen bands of seven M(r) were consistently and specifically recognized by IgE in the five-stage extracts by sera from prepatent and patent infections; only one such M(r) at 13.9 kDa, was described for IgG. A potentially diagnostic 31.9 kDa antigen band was identified on the IgE-specific EITB of D. immitis female extract and was shown to be recognized by IgE in sera from all infected dogs at all time points examined from 2 weeks until 1 year post-inoculation. Overall, IgE reactivity was more specific for D. immitis infections than IgG reactivity.  相似文献   

14.
Abstract Skin biopsy specimens from face, axilla, abdomen and thigh, mucocutaneous tissues from anus and vagina, and oral mucosa from six healthy Beagle dogs were examined for desmoplakin (Dsp) immunoreactivity using immunoblotting and immunohistochemical analysis. With immunoblotting using mouse antihuman Dsp 1 monoclonal antibody (DP2.17), a band was detected at 250 kDa in all the extracts as in normal human skin samples, although no band was detected at 210 kDa, suggesting that monoclonal antibody DP2.17 recognizes canine Dsp 1 but not Dsp 2. Moreover, the desmosome regions of all specimens were stained with DP2.17 using immunohistochemical analysis. From these results, DP2.17, developed for the examination of human skin, might be suitable for the investigation of Dsp-related skin disorders in dogs.  相似文献   

15.
The objective of this work was to describe the presence of osteopontin (OPN) in canine seminal plasma and sperm membranes. A pool of seminal plasma and sperm membrane extract from 30 dogs was used. Polyacrylamide electrophoresis gels were performed and the bands were transferred to nitrocellulose paper and Western blot was undertaken using an antibody anti-OPN. Two and 12 bands were marked in the seminal plasma (77.2 and 15.6 kDa) and sperm membrane extracts (70.6–26.6 kDa), respectively. However, from 12 marked bands in the sperm membrane extract, only three (46.4, 37.7 and 36.5 kDa) were strongly marked. We conclude that, seminal plasma and sperm membranes from dogs contain different isoforms of OPN; yet, further studies will be necessary to determine their function in this species.  相似文献   

16.
To investigate the predominance of Malassezia pachydermatis (M. pachydermatis) as a causative agent of canine otitis externa, ear cerumen samples were observed for adhesion of M. pachydermatis to the cornified epithelial cells by light and electron microscopes. The yeasts appeared not to adhere to the cornified epithelial cells directly, but they seemed to exist in the proximity of the epithelial cells with an electron opaque halo-like space around them. Polysaccharide and lipid staining techniques were conducted to identify the substances existing in that space. Lipid substances, not saccharides, were observed around the yeasts and the cornified epithelial cells. These results suggested that in the canine ear canal malassezia yeast attachment to the cornified epithelial cells is mediated by lipids.  相似文献   

17.
The purpose of this study was to investigate the diversity of yeast associated with the degree of canine seborrheic dermatitis (SD) by anatomical sites. Fifty-seven samples were divided as 17 healthy skin, 20 with primary seborrheic dermatitis (PSD), and 20 with secondary seborrheic dermatitis (SSD). Yeast isolation and characterization were carried out based on microscopical features and biochemical properties. DNA analysis at the internal transcribed spacer I of 26S rDNA region was utilized for species confirmation. Four species of yeast consisting Malassezia pachydermatis, Malassezia furfur, Candida parapsilosis and Candida tropicalis recovered from examined dogs. M. pachydermatis and C. parapsilosis were isolated from all dogs, but C. tropicalis and M. furfur were recovered from 3 healthy dogs and one diseased dog, respectively. The number of M. pachydermatis and C. parapsilosis in diseased dogs was higher than that of healthy specimens (P<0.01). High frequency and population size of C. parapsilosis were closely associated to PSD, while those of M. pachydermatis were associated with both PSD and SSD (P<0.01). C. parapsilosis were predominant at the perianal area. This study demonstrated the co-colonization of M. pachydermatis and C. parapsilosis in large amounts and frequency associated with stage of disease and anatomical site.  相似文献   

18.
An epidemiological investigation of 120 canine otitis externa cases in 1,370 dogs was done on the incidence rate, ear pinna shapes, breeds and their relationships. Eighty-five cases (12.6%) in 672 dogs with pendulous ears and 35 cases (5.0%) in 698 dogs with erect ears had otitis externa, and the difference between them was significant (P<0.05). Ninety-five auditory cerumen specimens were cultured for Malassezia pachydermatis (M. pachydermatis) and analyzed for concentrations of major fatty acids. Although rates of cases positive for M. pachydermatis in both ear pinna shapes were almost the same, i.e. 55.2% in the pendulous group and 53.6% in the erect group, the average total fatty acid level of the pendulous ear group was significantly (P<0.05) higher than that in the erect ear group after dismissing extraordinary levels in the Siberian husky. Isolated M. pachydermatis strains were examined for the effects of fatty acid supplementation on their growth. The majority of the strains utilized fatty acids and grew faster in fatty acid supplemented broth. These results suggest that M. pachydermatis, the predominant causative agent of canine otitis externa, prefers the auditory canal of dogs with lipid-rich earwax and grows fast, but growth strongly depends upon the canine breed.  相似文献   

19.
An indirect enzyme linked immunosorbent assay (ELISA) was developed and its diagnostic potential evaluated for rabbits infected by Trichophyton mentagrophytes. Within-run and between-run coefficient of variance varied from 2.3 to 7.7% and from 5.9 to 8.5%, respectively, indicating satisfactory reproducibility of the ELISA. There was no significant cross-reaction with antigens of Microsporum canis, Malassezia pachydermatis and Aspergillus fumigatus. The level of specific IgG to Trichophyton mentagrophytes was measured in sera of 25 11-week-old and 12 younger infected rabbits. There was no significant difference in the IgG level between 12 5-week-old infected rabbits and controls (p = 0.38). The antibody response was higher in 12 7-week-old rabbits compared with controls (p = 0.001). The IgG level in 25 11-week-old rabbits differed from the controls very significantly (p < 0.0001). Increased specific IgG in 11-week-old rabbits exhibited 96% sensitivity and 94% specificity. Predictive values of a positive and a negative test were 96 and 94%, respectively. Western immunoblotting associated three protein bands (21.5, 31, 44 kDa) with Trichophyton mentagrophytes infection.  相似文献   

20.
Lipid-dependent Malassezia species have recently been cultured from veterinary specimens. The identification of Malassezia species isolates from animals is important to clarify the epidemiology of these lipophilic yeasts. Malassezia species were cultured from the external ear canals of 63 out of 99 cats with otitis and 12 of 52 (23%) healthy control cats. The rate of isolation in affected animals versus controls was highly significant (P<0.01). Malassezia pachydermatis was isolated as a pure culture in 33 (45.2%) cats, associated with Malassezia globosa and Malassezia furfur in 20 (50%) and 17 (42.5%) animals, respectively. Three different species were isolated simultaneously in three cats (two cats with M pachydermatis, M globosa and M furfur, one subject with M pachydermatis, M furfur and Malassezia sympodialis). M globosa was isolated as the sole species in two animals. The present work confirms the presence of some lipid-dependent species of Malassezia in both healthy and otitic cats.  相似文献   

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