Inhibition of Dermatophilus congolensis by substances produced by bacteria found on the skin

Bacteria, isolated from the skins of clinically normal sheep, were tested for inhibitory activity against Dermatophilus congolensis grown in vitro. Out of 85 bacterial isolates, 19, mainly Bacillus spp., showed zones of inhibition when grown together with D. congolensis. The inhibitory activity was shown to be due to the metabolites released by the bacteria.     

盘羊刚果嗜皮菌的分离与鉴定

从疑似刚果嗜皮菌感染的盘羊皮肤病料中分离刚果嗜皮菌,并进行鉴定。将皮肤病料于28 ℃风干、粉碎,加无菌水浸泡15 d,静置,取上清加6%酵母膏液,60 ℃条件下30 min,混匀后连续10倍倍比稀释涂板,37 ℃培养48 h后挑取可疑菌落进行细菌形态学、培养特性、间接免疫酶标染色法鉴定,并根据GenBank上发表的刚果嗜皮菌16S rRNA序列设计特异性引物对分离株进行PCR扩增,得到与预期相符的606 bp基因片段,结果证实该盘羊分离株为刚果嗜皮菌。     

In vitro and in vivo inhibition of Dermatophilus congolensis by coagulase-negative antibiotic-producing staphylococci from pigs

When tested on solid media the growth of 19 Dermatophilus congolensis strains was inhibited by antibiotic-producing staphylococci isolated from pigs. Two strains, D congolensis D11 and D15, which were very sensitive to the producers and caused lesions of dermatophilosis in a mouse model, were further used to investigate the ability of the producers to inhibit lesion formation by the strains of D congolensis. The simultaneous application of the antibiotic-producing staphylococci and D congolensis suppressed formation of the lesions in the mouse.     

Humoral and cell-mediated immune response to crude antigens of Dermatophilus congolensis during experimental infection of rabbits.

Rabbits were infected with Dermatophilus congolensis and tested for humoral immune response by indirect haemagglutination and for cell-mediated immune response to crude antigens of D. congolensis. Lymphocyte transformation and macrophage migration inhibition assays were used as in vitro correlates of cell-mediated immune response while cutaneous delayed hypersensitivity was used in vivo. Endo-antigen and whole cell antigen were found to significantly induce cell-mediated immune response. In contrast, humoral responses were found to be more significantly induced by exo-antigen. A biphasic immune response was revealed by the lymphocyte transformation test.     

Use of a monoclonal antibody in the diagnosis of infection by Dermatophilus congolensis

A monoclonal antibody (McAb) to Dermatophilus congolensis was produced from murine hybridoma cultures and purified by affinity chromatography. Species specificity was demonstrated using indirect immunofluorescent staining; the McAb was shown to react with 10 D congolensis isolates but not with 10 Nocardia species isolates, a Rhodococcus and a Streptomyces species isolate. The McAb was used to demonstrate D congolensis in clinical material from confirmed bovine and ovine cases and presumptive equine cases of dermatophilosis by indirect immunofluorescent staining.     

Dual infection of a white-tailed deer by Dermatophilus congolensis and Alternaria alternata.

Infection by both Dermatophilus congolensis and Alternaria alternata was found in a 5 1/2-year-old, female white-tailed deer (Odocoileus virginianus). Encrusted lesions characteristic of dermatophilosis were observed on the hocks, flanks, and back. Giemsa-staining of smears of material from beneath the crusts revealed branching filaments, transversely and longitudinally divided into packets of coccoid cells typical of D congolensis. Hyphae morphologically consistent with those of A alternata were found in methenamine-silver- and hematoxylin-and-eosin-stained sections of tissue from the ears, flanks, and back. Nutrient agar cultures inoculated with tissue from an ear and hindlimb of the deer yielded, respectively, A alternata and D congolensis.     

Isolation of Dermatophilus congolensis from a cat

Dermatophilus congolensis was isolated from a cat with dermatitis. The isolate was sensitive to oxytetracyclin, streptomycin and penicillin but resistant to ampicillin, amoxicillin, gentamycin and cefoperazone.     

Dermatophilosis in Australian bearded lizards.

Dermatophilosis (Dermatophilus congolensis) was diagnosed in 3 Australian bearded lizards. Each lizard died of causes associated with stress and poor adaptability, although the lesions appeared to be regressing at the time of death.     

Serum and skin surface antibody responses in merino sheep given three successive inoculations with Dermatophilus congolensis

Three antigens prepared from different phases of the life cycle of Dermatophilus congolensis were used in an enzyme-linked immunosorbent assay to measure serum and skin surface antibody responses in sheep after a first, second and third inoculation with D. congolensis. After the first inoculation, a strong antibody response to the flagella, filament and soluble antigens was detected after 7-21 days in the sera from sheep that were regularly biopsied; the antibody response at the skin surface was detected 28-42 days after inoculation, when the lesions were resolving. Strong anamnestic responses were detected in the serum of sheep that were biopsied and some of the nonbiopsied sheep after the second and third inoculations, but the skin surface antibody response at these times was variable.     

A comparison of extracted proteins of isolates of Dermatophilus congolensis by sodium dodecyl sulphate-polyacrylamide gel electrophoresis and Western blotting.

Antigenic diversity within a collection of 18 isolates of Dermatophilus congolensis from different Continents was examined by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and by Western blotting with sera from cattle with clinical dermatophilosis using whole cell extracts obtained by three methods and one extract of extracellular products of D. congolensis. One of the methods involving the release of a lysostaphin-solubilized protein (LSP) of whole cells of D. congolensis revealed a number of discrete and easily-identifiable bands in SDS-PAGE which were found suitable for characterizing protein patterns and was, therefore, subsequently used for a comparative analysis of the proteins of all the D. congolensis isolates. Six electropherotypes (ET) of D. congolensis were identified among the 18 isolates using the protein profiles based on the presence of four protein bands at Molecular weights (MW) 62, 28, 17.4 and 16.4 kDa. The ETs were found among isolates from different animal species and from different sources with ET1 consisting of three bovine and two equine isolates; ET2, two bovine and three ovine isolates; ET3, two bovine isolates; ET4, two bovine isolates; ET5, one bovine and one ovine isolates and ET6, two bovine isolates. Immunoblotting of the extracts of D. congolensis isolates with sera from cattle with clinical dermatophilosis infection demonstrated protein bands of MW ranging from 9 kDa to 188 kDa. Sera from chronic dermatophilosis infection demonstrated a 28 kDa protein which was immunodominant in the LSP extracts of all the 18 isolates of D. congolensis tested while sera from mild infections demonstrated mainly the 62 kDa protein in the same extracts. However, many protein bands were demonstrated in surface membrane (TSMP) and extracellular protein extracts with sera from only mildly infected animals. The protein patterns observed in all isolates of D. congolensis revealed global antigenic similarities and distinct differences among isolates which could not be associated with either geographic, climatic or host factors. Also sera from infected animals from endemic regions of dermatophilosis could not differentiate isolates of D. congolensis. This suggests the possibility that such sera must have come from animals that had been infected by a multitude of D. congolensis strains present in the herd environment and strains an animal could have come across during the 'ritual' annual cross-country migration of the cattle herds.     

Serodiagnosis of Dermatophilus congolensis infection by counterimmunoelectrophoresis

Sixty-one sera from animals that had contact with Dermatophilus congolensis were examined by comparing three serological methods; counterimmunoelectrophoresis, passive haemagglutination, and agar gel diffusion, and by using four different antigenic extracts of D congolensis. The counterimmunoelectrophoresis was the most satisfactory of the methods having been found to be specific and sensitive, easy to perform and suitable for screening large numbers of samples. It was also found to have a higher antibody detection rate (82.2 per cent) than the other methods thus making it suitable for seroepidemiological surveys. It was found to be capable of detecting multiple antibodies and also revealed dissimilarities among the different antigenic extracts. The cellular antigens of D congolensis were found to detect antibody in more sera than the extracellular antigen; the cell wall extract proved to be the most satisfactory of all, detecting antibody from the largest number of sera compared to the other extracts in all the three serological tests.     

Hemolytic interactions of Dermatophilus congolensis.

The strains of Dermatophilus congolensis grew on blood agar with washed sheep erythrocytes with marked total hemolysis. In testing for hemolytic interactions they gave a significant synergistic effect of a characteristic shape with Rhodococcus equi and Streptococcus agalactiae, whereas with Staphylococcus aureus producing beta hemolysin and with Staphylococcus aureus producing delta hemolysin a simultaneous synergistic as well as antagonistic effect were observed. First of all a conspicuous inhibition of in the beta hemolysin zone began and then the hemolytic effect of D. congolensis was enhanced. A similar double reaction was also observed with Listeria ivanovii. With delta hemolysin there was an inhibition of the hemolytic effect of D. congolensis and at the same time a synergistic effect could be observed. Also D. congolensis gave a weak synergistic effect with Micrococcus lylae and Listeria monocytogenes, and a further weak antagonistic effect with alpha hemolysin of Staphylococcus aureus, Staphylococcus hyicus, Staphylococcus chromogenes and Micrococcus luteus. No interaction of D. congolensis was established with Corynebacterium pseudotuberculosis.     

Oral dermatophilosis in a cat: a case report

An oral lesion in a cat caused by the actinomycete Dermatophilus con-golensis is described. Multiple granulomata were present in biopsy material taken from a tongue lesion. After surgical excision and daily penicillin therapy the condition regressed.
Résumé. Une lésion oralle dans un chat, causée par l'actinomycète Dermatophilus congolensis est décrit. Des granulomes multiples étaient présentes dans la substance de la biopsie, prise de la lésion de la langue. Après l'excision chirurgicale et la thérapie journalière de pénicilline, la condition décroissait.
Zusammenfassung. Man beschreibt eine orale Verletzung in einer Katze, die durch den Strahlenpilz Dermatophilus congolensis , verursachet wird. Die multipelen Granulomata waren im Stoff von der Biopsie, Welches von der Verletzung der Zunge, genommen wird. Der Zustand klang ab, nach der chirurgischen Exicision und nach der täglichen Heilung von Penizillin.     

Relationship between chronic ovine dermatophilosis and levels of T6 lymphocyte antigen staining in peripheral blood mononuclear cells.

Quantification of a T6-lymphocyte antigen in peripheral blood mononuclear cells of sheep was used to select 15 from 48 one year old Merino ewes not previously exposed to Dermatophilus congolensis infection. These sheep were compared in response to challenge with D. congolensis zoospores and levels of T-6 lymphocyte antigen in peripheral blood mononuclear cells with 15 Merino ewes of similar age and strain from a different site that had been treated and recovered from chronic dermatophilosis. The T-6 lymphocyte antigen levels were significantly lower in the chronic dermatophilosis sheep and they developed significantly more severe lesions than the selected, previously unexposed sheep despite the former sheep having high serum antibody levels to D. congolensis. Measurement of the fleece characteristics, wax and suint concentration showed no differences between the groups that might have explained the considerable differences in their susceptibility to dermatophilosis.     

Cellular responses in the skin of merino sheep to repeated inoculation with Dermatophilus congolensis

The cellular response in the skin of Merino sheep was examined after three successive inoculations with Dermatophilus congolensis. There was a massive neutrophil influx into the infected epidermis and underlying dermis at 4-10 days after the first inoculation. A lymphocyte-macrophage response occurred at 10-12 days, followed by a plasma cell response at 14-38 days. Resolution of skin lesions after the first inoculation corresponded to the time when the plasma cell response in the skin was most intense. A second inoculation with D. congolensis, 70 days after the first, failed to produce skin lesions typical of dermatophilosis. Typical lesions of dermatophilosis did develop after a third inoculation of the same sheep 140 days after the first inoculation, but the lesions resolved in most sheep within 13 days. Dermatophilosis did not develop in some of these sheep at sites inoculated with 100-1000-fold lower infective doses of D. congolensis, whereas control sheep did develop lesions.     

Temporal changes in the populations of immune cells at the site of experimental Dermatophilus congolensis infection in mice and sheep

Abstract The temporal patterns of dermal immune cell influx were compared in mice and sheep, species reputedly resistant and susceptible, respectively, to infection with Dermatophilus congolensis. In both species, the response involved early mast cell degranulation, vasodilatation and an influx of dendritic cells which accumulated and apparently differentiated beneath the infected epidermis. A concomitant dermal invasion by neutrophils and T and B lymphocytes led to epidermal infiltration, particularly by neutrophils and thence to the formation of the surface scab. Hypertrophy of the epidermis also indicates keratinocyte involvement in the host response. The duration of the response, however, was considerably shorter in the mouse (about 5 days) and B cells' were the predominant lymphocyte under and adjacent to the lesion. During the more protracted response in the sheep (> 21 days), T cells, including T19 antigen +γδ T cells, outnumbered B cells. Résumé— Des modifications dans les populations des cellules immunes dans le derme sont comparées chez la souris et le mouton, espèces respectivement résistantes et sensibles à Dermatophilus congolensis. Dans ces deux espèces, la réponse fait intervenir d'abord la dégranulation des mastocytes, la vasodilatation et l'arrivée de cellules dendritiques qui s'accumulent et se différencient apparemment sous l'épiderme infecté. Une invasion concomittente dermique par des neutrophiles, des lymphocytes T et B est observée et aboutit à l'infiltration épidermique, particulièrement par les neutrophiles et pour cette raison, à la formation de croûtes superficielles. L'hypertrophie de l'épiderme indique aussi l'implication du kératinocyte dans la réponse de l'hôte. Cependant, la durée de cette réponse, est bien plus courte chez la souris (environ 5 jours) et les lymphocytes B sont les lymphocytes prédominants, sous-jacents à la lésion. Chez le mouton, la réponse est prolongée (plus de 21 jours) et les lymphocytes T, incluant les lymphocytes CD 19 et γδ surpassent en nombre les lymphocytes B. [Sasiak, A.B., Lloyd, D.H., McEwan Jenkinson, D., Kitson, S., Elder, H. Y. Temporal changes in the populations of immune cells at the site of experimental Dermatophilus congolensis infection in mice and sheep (Particularités de la population cellulaire immunitaire aux sites d'infections expérimentales à Dermatophilus congolensis chez la souris et le mouton). Veterinary Dermatology 1996; 7 : 59–66.] ResumeAn Se compararon los patrones temporales de llegada de células inmunitarias en el ratón y en la oveja, especies consideradas resistentes y susceptibles, respectivamente a la infección por Dermatophilus congolensis. En ambas especies la respuesta implicó degranulación temprana de mastocitos, vasodilatación y llegada de células dendríticas que se acumulaban y aparentemente diferenciaban bajo la epidermis infectada. La invasión dérmica concomitante por netrófilos y linfocitos T y B llevó a una infiltración epidérmica, especialmente por neutrófilos y consecuentemente a la formación de una costra superficial. La hipertrofi de la epidermis también indica la implicación de queratinocitos en la respuesta del huésped. Sin embargo, la duración de la respuesta fue considerablemente más corta en el ratón (unos 5 dias) y los linfocitos B fueron la célula predominante en la zona adyacente a la lesión y debajo de ella. Durante la respuesta más prolongada en la oveja (> 21 días), las células T, incluyendo el antigeno T19, células T γδ, superaban en número las células B. [Sasiak, A.B., Lloyd, D.H., McEwan Jenkinson, D., Kitson, S., Elder, H. Y. Temporal changes in the populations of immune cells at the site of experimental Dermatophilus congolensis infection in mice and sheep (Alteraciones temporales en las poblaciones de celulas inmunitarias en zonas cutaneas de infeccion experimental por Dermatophilus congolensis en raton y en la oveja). Veterinary Dermatology 1996; 7 : 59–66.] Zusammenfassung— Die temporären Muster des Influx dermaler Immunzellen wurden bei Maus und Schaf verglichen, Tierarten, die angeblich resistent und empfänglich speziell für die Infektion mit Dermatophilus congolensis sind. Bei beiden Spezies bezog die Reaktion eine frühe Mastzelldegranulation ein, weiterhin Vasodilatation und einen Influx dendritischer Zellen, die sich unter der infizierten Epidermis anhäuften und offensichtlich differenzierten. Eine begleitende Invasion der Dermis durch Neutrophile, T- und B-Lymphozyten führte zu einer epidermalen Infiltration, besonders durch Neutrophile und von dort an zur Bildung von Oberflächenschorf. Die Hypertrophic der Epidermis zeigt außerdem eine Beteiligung der Keratinozyten bei der Antwort des Wirtstieres. Die Dauer der Reaktion jedoch war bei der Maus beträchtlich kürzer (etwa 5 Tage) und die B-Zellen waren die vorherrschenden Lymphozyten unter und angrenzend an die Läsion. Während der mehr protrahierten Reaktion beim Schaf (> 21 Tage) übertrafen die T-Zellen einschließlich T19-Antigen und gamma-delta-T-Zellen die Anzahl der B-Zellen. [Sasiak, A. B., Lloyd, D. H., McEwan Jenkinson, Kitson, S., Elder, H. Y. Temporal chnges in the populations of immune cells at the site of experimental Dermatophilus congolensis infection in mice and sheep (Temporäre Veränderungen in den Populationen von Immunzellen an der Stelle experimenteller Infektionen mit Dermatophilus congolensis bei Maus und Schaf). Veterinary Dermatology 1996; 7 : 59–66.]     

Survival of Dermatophilus congolensis in tropical clay soils submitted to different water potentials.

The survival of a rifampicin-resistant mutant of Dermatophilus congolensis in vertisol and oxisol soils from Guadeloupe and in their constitutive clays was studied using a pneumatic device for controlling water potentials (pF). Experiments were carried out at two pF values simulating the wet season and the dry season. Survival time depended on the type of soil and its water content. Organic matter had a protective effect on the microorganism in oxisol but not in vertisol. The pathogenicity of D. congolensis was preserved in the soils which could therefore act as temporary reservoirs of this pathogen. Long-term survival of this organism in soils mixed with water suggests that ponds and dipping tanks may constitute sources of infection for cattle.     

Immunodominant antigens of zoospores from ovine isolates of Dermatophilus congolensis.

Zoospores of Dermatophilus congolensis were analysed by SDS-PAGE and western blotting. The electrophoretic profiles of zoospores from 13 isolates of D. congolensis were similar but not identical when stained with Coomassie blue or silver. Immunodominant polypeptides with apparent molecular masses of 76 and 31 kDa were identified in western blots of 13 of 13 and 12 of 13 isolates respectively of D. congolensis reacted with hyperimmune, ovine, antizoospore sera. Identical immunodominant polypeptides were observed in western blots reacted with sera obtained from naturally infected sheep. Initial characterisation of the 76 and 31 kDa polypeptides indicated that they were probably surface exposed because (i) antibodies eluted from the surface of live zoospores after adsorption of hyperimmune antizoospore serum, reacted principally against the 76 and 31 kDa subunit polypeptides in western blots, (ii) adsorption of hyperimmune antizoospore serum with live zoospores resulted in significant diminution of reactivity against both the 76 and 31 kDa polypeptides in western blots, (iii) indirect fluorescent immunostaining of zoospores with antiserum prepared against gel-purified 76 kDa polypeptide, resulted in intense staining of the zoospore outer coat. Immuno-gold electron microscopy of negatively stained zoospores with antiserum prepared against gel-purified 31 kDa polypeptide identified this antigen as a flagella subunit.     

Vaccination against ovine dermatophilosis

Zoospore, filamentous and soluble antigens were prepared from Dermatophilus congolensis and examined for their ability to protect sheep from challenge with D. congolensis zoospores. In 1 experiment, sheep were vaccinated with Antigens A, B and C. The number of sheep protected in the group vaccinated with Antigen B was greater (P less than 0.05) than that in the unvaccinated group after challenge. The group vaccinated with Antigen B had a higher antibody response (P less than 0.05) to Antigen B than to Antigen A or C. In a second experiment, 2 groups of sheep were vaccinated with Antigen B. All sheep in this study developed lesions after challenge, but those on the vaccinated sheep were less severe (P less than 0.05) than those on the unvaccinated sheep. The antibody response to Antigen A, 28 days after vaccination, was higher (P less than 0.05) than the response to Antigen B.     

Evaluation of vaccines against ovine dermatophilosis

Intradermal vaccination of live crude filaments (vaccine A) was compared with a vaccine (vaccine B) consisting of a 45 kD zoospore protein and mucoid material coating filaments in its ability to protect sheep from experimental Dermatophilus congolensis infection. Fourteen and 21 days after challenge, vaccine A sheep had fewer lesions (P less than 0.001) than the vaccine B sheep. The lesions on the vaccine A sheep were also less severe 14 and 21 days after challenge (P less than 0.05, P less than 0.01 respectively). In a second study, vaccine A was assessed for its ability to protect against natural challenge. Ten weeks after contact with sheep with active and generalised dermatophilosis no difference was found between the number of lesions present on the vaccine A and unvaccinated sheep and no differences were found in the number of sheep in each group with active lesions.