Multi-azole-resistant strain of Malassezia pachydermatis isolated from a canine Malassezia dermatitis.

In the case presented here, we describe the isolation of an azole-resistant strain of M. pachydermatis from a canine Malassezia dermatitis. The isolate (NUBS18001) from this case exhibited a minimum inhibitory concentration (MIC) of 320 µg/ml to itraconazole (ITZ) by broth microdilution (BM) assay, >32 µg/ml to ITZ by E-test, and >32 µg/ml to KTZ by E-test. Synergistic effects between FK506 and ITZ in the azole-resistant strain was evaluated using the microdilution checker-board method. The ITZ-resistant strain exhibited MICs of 320 µg/mL of ITZ alone and 5 µg/ml of FK506 alone; the addition of FK506 attenuated the ITZ MIC to 2.5 µg/ml, yielding an ITZ FICI value of 0.507. This result suggested that the combination of ITZ and FK506 exerted an additive effect against the ITZ-resistant strain. To understand the other mechanism inferred to be present in our multi-azole-resistant strain, we sequenced the ERG11 gene from this isolate, and detected missense mutations (A412G and C905T) in the sequence of the ERG11 open reading frame (ORF). To the best of our knowledge, this work is the first report that a multi-azole-resistant M. pacydermatis strain contains mutations in ERG11.

Daily very low UV dose exposure enhances adaptive immunity, compared with a single high-dose exposure. Consequences for the control of a skin infection.

Ultraviolet radiation (UVr) promotes several well-known molecular changes, which may ultimately impact on health. Some of these effects are detrimental, like inflammation, carcinogenesis and immunosuppression. On the other hand, UVr also promotes vitamin D synthesis and other beneficial effects. We recently demonstrated that exposure to very low doses of UVr on four consecutive days [repetitive low UVd (rlUVd)] does not promote an inflammatory state, nor the recruitment of neutrophils or lymphocytes, as the exposure to a single high UV dose (shUVd) does. Moreover, rlUVd reinforce the epithelium by increasing antimicrobial peptides transcription and epidermal thickness. The aim of this study was to evaluate the adaptive immune response after shUVd and rlUVd, determining T-cell and B-cell responses. Finally, we challenged animals exposed to both irradiation procedures with Staphylococcus aureus to study the overall effects of both innate and adaptive immunity during a cutaneous infection. We observed, as expected, a marked suppression of T-cell and B-cell responses after exposure to an shUVd but a novel and significant increase in both specific responses after exposure to rlUVd. However, the control of the cutaneous S. aureus infection was defective in this last group, suggesting that responses against pathogens cannot be ruled out from isolated stimuli.

Plasma Pharmacokinetics of Polyphenols in a Traditional Japanese Medicine, Jumihaidokuto, Which Suppresses Propionibacterium acnes-Induced Dermatitis in Rats.

Most orally administered polyphenols are metabolized, with very little absorbed as aglycones and/or unchanged forms. Metabolic and pharmacokinetic studies are therefore necessary to understand the pharmacological mechanisms of polyphenols. Jumihaidokuto (JHT), a traditional Japanese medicine, has been used for treatment of skin diseases including inflammatory acne. Because JHT contains various types of bioactive polyphenols, our aim was to clarify the metabolism and pharmacokinetics of the polyphenols in JHT and identify active metabolites contributing to its antidermatitis effects. Orally administered JHT inhibited the increase in ear thickness in rats induced by intradermal injection of Propionibacterium acnes. Quantification by LC-MS/MS indicated that JHT contains various types of flavonoids and is also rich in hydrolysable tannins, such as 1,2,3,4,6-penta-O-galloyl glucose. Pharmacokinetic and antioxidant analyses showed that some flavonoid conjugates, such as genistein 7-O-glucuronide and liquiritigenin 7-O-glucuronide, appeared in rat plasma and had an activity to inhibit hydrogen peroxide-dependent oxidation. Furthermore, 4-O-methylgallic acid, a metabolite of Gallic acid, appeared in rat plasma and inhibited the nitric oxide reaction. JHT has numerous polyphenols; it inhibited dermatitis probably via the antioxidant effect of its metabolites. Our study is beneficial for understanding in vivo actions of orally administered polyphenol drugs.

Antifungal activity of berberine hydrochloride and palmatine hydrochloride against Microsporum canis -induced dermatitis in rabbits and underlying mechanism.

BACKGROUND: Phellodendron amurense, exhibits antifungal activity mainly by bioactive components including berberine hydrochloride and palmatine hydrochloride. This study was conducted to evaluate the antifungal effects of berberine hydrochloride, palmatine hydrochloride, and a mixture of both substances against Microsporum canis in vivo and in vitro. METHODS: The minimal inhibitory concentrations (MICs) of monomers and clotrimazole were determined using 1.5 % tryptic soy agar. The effects of these drugs on Microsporum canis growth was detected by determining dry weight. Transmission electron microscopy (TEM) was performed to observe the effect of chemicals on cell ultrastructure. Differential mRNA expressions of eight genes of M. canis treated with berberine or palmatine or their combination at different time points were determined by real-time PCR. NADH enzyme concentration was also detected. Clinical evaluation via in-vivo antifungal assay was also performed. Skin histology PAS staining was also carried out. RESULTS: Results showed that MICs of berberine, palmatine and clotrimazole were 1, 1, and 0.015 mg/mL, respectively. No significant difference was observed among the growth curves of the three groups before 18 h was reached. TEM showed that these drugs could destroy the cell membrane and organelles of M. canis at different time points. After 30 h of incubation, relative mRNA expressions of the genes in the combined group were significantly higher than those in the other groups including the clotrimazole group (P < 0.05); Palmatine initially induced the mRNA up-regulation of PGAL4, FSH1, PQ-LRP, NADH1 and NDR in M. canis; by contrast, berberine maintained a high expression level of these genes to shorten fungal life cycle and eradicate M. canis. Clinical results showed that combined treatment was more effective than single administration of each monomer or clotrimazole. Hence, berberine mixed with palmatine significantly elicited antifungal activities and could be used to treat M. canis in rabbits. CONCLUSION: These results provide a comprehensive view of the mechanism of berberine and palmatine in anti-M. canis activity.

Clinical and mycological evaluation of an herbal antifungal formulation in canine Malassezia dermatitis.

OBJECTIVE: Malassezia pachydermatis is a common cause of more widespread dermatitis in dogs (CMD). Recurrences are common, and this disorder can be very troubling for both dogs and for the pet owner. MATERIAL AND METHODS: The treatment of 20 dogs affected by dermatitis due to M. pachydermatis, with Malacalm(®), a commercially available mixture consisting of essential oils (Citrus aurantium 1%, Lavandula officinalis 1%, Origanum vulgare 0.5%, Origanum majorana 0.5%, Mentha piperita 0.5% and Helichrysum italicum var. italicum 0.5%, in sweet almond oil and coconut oil) is reported. The effectiveness of the whole mixture, of component essential oils and of their more represented compounds against clinical isolates was evaluated by a microdilution test. Twenty animals were topically administered the mixture twice daily for 1 month. Ten animals were treated with a conventional therapy based on ketoconazole 10mg/kg/day and chlorhexidine 2% twice a week for 3 weeks. At the end of both treatments animals significantly improved their clinical status. Adverse effects were never noticed. Follow-up visit performed on day 180th allowed to observe a recurrence of clinical signs in all the subjects treated conventionally, while not significant clinical changes were referred in dogs treated with Malacalm(®). The overall MIC value of Malacalm(®) was 0.3%. O. vulgare showed the lowest minimum inhibitory concentrations (MIC), being active at 0.8%, followed by M. piperita (1%), O. majorana (1.3%), C. aurantium (2%) and L. officinalis (4%) while H. italicum did not yield any antimycotic effect up to 10%. Active major compounds were thymol, carvacrol, p-cymene, 1,8-cineol, limonene and menthol. CONCLUSION: The phytotherapic treatment achieved a good clinical outcome, and no recurrence of skin disorders on day 180th was recorded. This herbal remedium appeared to be a safe tool for limiting recurrences of CMD.

Staphylococcus aureus infection of the feet following fish pedicure.

We report a case of Staphylococcus aureus infection of the feet that appeared after a "fish pedicure" (immersion of the feet in a tank with the fish Garra rufa, that nibbles off dead skin). Clinical picture was characterized by maceration, purulent discharge, scales, crusts, itching and burning sensation. Bacteriological examinations were positive for Staphylococcus aureus. Mycological examinations were negative. The patient was successfully treated with ciprofloxacin. Only one case of skin foot infection after fish pedicure was reported so far. Fish pedicure can be a potentially dangerous procedure in immunocompromised or diabetic patients.

Feed enzyme supplementation does not ameliorate Foot Pad Dermatitis in broiler chickens fed on a corn-soyabean diet.

1. Foot Pad Dermatitis (FPD) can be a serious health, quality and welfare problem in poultry production, with a significant affect on the economics of production. The physico-chemical properties of (NSP) have been correlated with increases in excreta viscosity and adherence, and hence FPD in broiler chickens. 2. Two broiler experiments were conducted to evaluate the effects of different dietary non-starch polisaccharide (NSP) applications to corn-soyabean based diets in replicated floor pens. In addition to live performance, the incidence and severity of FPD was scored and, in experiment 2, intestinal viscosity and ammonia volatilization were measured. 3. Live performance did not vary with the different treatments in either experiment and the incidence and severity of FPD, and ammionia volatisation in experiment 2, were unaffected. However, birds reared on enzyme supplemented diets had lower (P < 0·001) intestinal viscosity levels in experiment 1. 4. In this study with corn-soyabean meal based diets, dietary enzyme supplementation had no affect on the incidence and severity of FPD. The lack of an association between the excreta viscosity and FPD is attributed to the multifactorial etiology of this condition.

Ulcerative dermatitis in C57BL/6 mice lacking stearoyl CoA desaturase 1.

Ulcerative dermatitis (UD) is a common cause of morbidity and euthanasia in mice with a C57BL/6 (B6) background. The purposes of the current study were to determine whether UD lesions could be reliably produced in B6 mice lacking stearoyl-CoA desaturase 1 (SCD1(-/-) mice), to ascertain whether the UD lesions in SCD1(-/-) mice were similar to those found in other B6 mice, and to characterize the cell invasion phenotype of Staphlococcus xylosus cultured from the lesions. S. xylosus isolates from the environment and human skin were used as controls. SCD1(-/-) (n = 8 per group) and nontransgenic B6 control mice (n = 22 mice pooled from 3 groups that received different concentrations of conjugated linoleic acid) were fed standard rodent chow or a semipurified diet (NIH AIN76A) for 4 wk. Samples from other B6 mice with UD (field cases; n = 7) also were submitted for histology and culture. All of the SCD1(-/-) mice developed UD lesions by 4 wk on NIH AIN76A. None of SCD1(-/-) fed standard rodent chow and none of the wildtype B6 mice fed NIH AIN76A developed UD. Supplementation with conjugated linoleic acid did not affect ulcerogenesis. UD lesions in SCD1(-/-) mice and field cases were grossly and histologically similar. S. xylosus was isolated from SCD1(-/-) mice with UD (71%) and field cases of UD (43%). These isolates were the most cell-invasive, followed by the environmental isolate, and finally the human skin isolate. Our results provide a basis for further pathologic and clinical study of UD.

Inhibitory effect of formulated lemongrass shampoo on Malassezia furfur: a yeast associated with dandruff.

Lemongrass (Cymbopogon citratus Stapf) has been used in cooking and in many traditional medicines; the essential oil contains citral as a major constituent. This study evaluated the antifungal activity of lemongrass oil against Malassezia furfur, an opportunistic yeast associated with dandruff, by using a broth dilution assay. From the minimum fungicidal concentration (MFC) obtained, the oil was then incorporated at different percentages into shampoo formulations. The formulated shampoos were kept at room temperature (28 degrees-30 degrees C) and under accelerated condition (45 degrees C). At the end of the first and sixth weeks, after preparation, all formulations were tested again and the appearance was recorded. Selection of an appropriate formula was based on antifungal activity against M. furfur, the physical appearance, the chemical properties and stability of the formula. Two percent lemongrass oil shampoo provided the required qualities necessary for commercial use. After being kept for 6 weeks at 28 degrees-30 degrees C and 45 degrees C, this formulated shampoo gave MFCs against M. furfur of 75 microl/ml and 18.75 microl/ml, respectively.

Antibacterial and dermal toxicological profiles of ethyl acetate extract from Crassocephalum bauchiense (Hutch.) Milne-Redh (Asteraceae).

BACKGROUND: The emergence in recent years of numerous resistant strains of pathogenic bacteria to a range of formerly efficient antibiotics constitutes a serious threat to public health. Crassocephalum bauchiense, a medicinal herb found in the West Region of Cameroon is used to treat gastrointestinal infections as well as liver disorders. The ethyl acetate extract from the leaves of C. bauchiense was evaluated for its antibacterial activity as well as acute and sub-acute toxicities. METHODS: The plant extract was prepared by maceration in ethyl acetate. Its phytochemical screening was done by standard methods. The broth microdilution method was used to evaluate the in vitro antibacterial activity. The in vivo antibacterial activity of a gel formulation (0.05, 1 and 2% w/v) of this extract was evaluated using a Staphylococcus aureus-induced dermatitis in a murine model. Selected haematological and biochemical parameters were used to evaluate the dermal sub-acute toxicity of the extract in rats. RESULTS: Phytochemical screening of the C. bauchiense extract revealed the presence of alkaloids, phenols, tannins and sterols. In vitro antibacterial activities were observed against all the tested microorganisms (MIC = 0.04-6.25 mg/ml). Formulated extract-gel (2% w/v) and gentamycin (reference drug) eradicated the microbial infection after five days of treatment. A single dermal dose of this extract up to 32 g/kg body weight (bw) did not produce any visible sign of toxicity. Also, daily dermal application of the C. bauchiense extract gel formulation for 28 days did not show any negative effect, instead some biochemical parameters such as alanine aminotransferase (ALT and AST), low density lipoprotein (LDL), high density lipoprotein (HDL) and triglycerides were significantly (p < 0.05) affected positively. CONCLUSION: These results indicate that the C. bauchiense ethyl acetate extract can be used safely for the treatment of some bacterial infections.

Inhibitory effects of Fortunella japonica var. margarita and Citrus sunki essential oils on nitric oxide production and skin pathogens.

A number of essential oils from citrus peels are claimed to have biological activities. Citrus peel, called 'Jin-Pi', is used in traditional medicine for digestion, severe cold, and fever. However, the antibacterial activities against skin pathogens and anti-inflammatory effects of the essential oils of Citrus sunki (JinGyul) and Fortunella japonica var. margarita (GumGyul) have not yet been described. Therefore, in this study, the essential oils of the citrus species C. sunki (CSE) and F. japonica var. margarita (FJE), both native to the island of Jeju, Korea, were examined for their anti-inflammatory and antimicrobial activities against skin pathogens. Four human skin pathogenic microorganisms, Staphylococcus epidermidis CCARM 3709, Propionibacterium acnes CCARM 0081, Malassezia furfur KCCM 12679, and Candida albicans KCCM 11282, were studied. CSE and FJE exhibited strong antimicrobial activity against most of the pathogenic bacteria and yeast strains that were tested. Interestingly, CSE and FJE even showed antimicrobial activity against antibiotic-resistant S. epidermidis CCARM 3710, S. epidermidis CCARM 3711, P. acnes CCARM9009, and P. acnes CCARM9010 strains. In addition, CSE and FJE reduced the lipopolysaccharide (LPS)-induced secretion of nitric oxide (NO) in RAW 264.7 cells, indicating that they have anti-inflammatory effects. We also analysed the chemical composition of the oils by gas chromatography-mass spectrometry (GC-MS) and identified several major components, including dl-limonene (68.18%) and beta-myrcene (4.36%) for CSE, and dl-limonene (61.58%) and carvone (6.36%) for FJE. Taken together, these findings indicate that CSE and FJE have great potential to be used in human skin health applications.

Designing a successful antimicrobial treatment against Devriesea agamarum infections in lizards.

Dermatitis caused by Devriesea agamarum poses a major health problem for the captive maintenance of several desert lizard species. This study was conducted to determine the optimal antimicrobial treatment to eliminate D. agamarum infections from lizards. First, the in vitro susceptibility of 42 D. agamarum isolates was determined for 10 different antimicrobial agents using an agar dilution method. In none of the isolates acquired antimicrobial resistance was demonstrated. Then, two intramuscular treatment protocols using either enrofloxacin or ceftiofur were tested in bearded dragons (Pogona vitticeps) experimentally infected with a D. agamarum strain showing a MIC of 2 microg/ml for enrofloxacin and 0.12 microg/ml for ceftiofur. While D. agamarum could no longer be isolated after 17-18 days of ceftiofur administration, enrofloxacin administration and sham treatment failed in clearing the infection after 27 days of treatment. Based on these results, intramuscular injection of ceftiofur at 5 mg/kg BW q24h was used to treat naturally and clinically infected Uromastyx lizards. This resulted in marked clinical improvement and clearance of infection after 12 days on average. In conclusion, intramuscular administration of ceftiofur at 5 mg/kg BW q24h eliminates D. agamarum in lizards, resulting in clinical cure.

Genetic heterogeneity among strains of Treponema phagedenis-like spirochetes isolated from dairy cattle with papillomatous digital dermatitis in Japan.

Papillomatous digital dermatitis (PDD) is an infectious foot disease of cattle that is prevalent throughout the world. Although it has been prevalent in Japan since the first case was reported in 1992, full epidemiological and bacteriological examinations have not been conducted. We collected 91 lesions of PDD from 80 dairy cattle on 12 farms in eight regions of Japan to isolate the spirochetes that are frequently detected in lesions. We isolated 40 strains of spirochetes from 24 cattle (30.0%) by a simple two-step culture technique, in which the biopsy samples were incubated at 4 degrees C for 48 to 72 h in an enrichment broth supplemented with antibiotics, which improved the rate of isolation, and then inoculated on selective agar plates. All spirochetes examined were catalase positive and oxidase negative and showed weak beta-hemolytic activity. Enzyme activities were identical to those of Treponema phagedenis ATCC 27087. Sequencing of the 16S rRNA gene showed that all strains isolated had >99% identity to those of the T. phagedenis type strain and of T. phagedenis-like strains isolated from PDD lesions in the United States and Europe. Pulsed-field gel electrophoresis and PCR-based random amplified polymorphism DNA methods revealed considerable diversity among strains isolated not only from different cattle but also from the same individuals. These findings may provide further evidence for the role of these treponemes in the pathogenesis of persistent PDD.

Mycobacterium intermedium granulomatous dermatitis from hot tub exposure.

Nontuberculous mycobacteria, which are widespread in the environment, frequently cause opportunistic infections in immunocompromised patients. We report the first case of a patient with chronic granulomatous dermatitis caused by a rarely described organism, Mycobacterium intermedium. The infection was associated with exposure in a home hot tub.

Acute urethritis and arthritis-dermatitis syndrome due to Neisseria meningitidis.

Acute urethritis and arthritis-dermatitis syndrome after sexual contact are often assumed to be caused by Neisseria gonorrhoeae. We report a case of arthritis-dermatitis syndrome in a 32-year-old man who presented with generalized maculopapular and petechial skin lesions and polyarthritis. Acute urethritis developed 1 week after oro-genital sexual contact with a sex worker about 3 weeks before admission. No pathogen was found on smear of urethral discharge and skin lesions, but Gram-negative diplococci were noted in joint fluid, and blood culture yielded N. meningitidis. His condition improved gradually after repeated arthrocentesis and antibiotic therapy with ceftriaxone followed by ciprofloxacin. Oro-genital contact is a transmission route for N. meningitidis infection manifesting as acute urethritis and arthritis-dermatitis syndrome.

Propionibacterium acnes lipase in seborrheic dermatitis and other skin diseases and Unsei-in.

We examined Propionibacterium acnes lipase in skin diseases and Unsei-in. Butyric acid production in axillary seborrheic dermatitis (ASD) was higher than in other dermatitis, and that in acne vulgaris (AV) was significantly higher than in controls. P. acnes lipase is the pathogenic factor in AV and fatty acids produced by lipase might be the pathogenic factor in ASD. Unsei-in suppressed P. acnes lipase probably because some ingredients have antimicrobial and anti-inflammatory activities.

Candid-B cream in the treatment of candidiasis with inflammatory dermatoses--National Study Group.

Physicians (n = 84) across the country prescribed candid-B cream (clotrimazole 1% + beclomethasone dipropionate 0.025%) on 822 patients suffering from candidiasis with inflammatory diseases to evaluate the efficacy and safety of the combination. The results showed reduction in severity was more than 80% for all symptoms/signs except scaling and lichenification where the reduction was 76.05% and 66.03% respectively. Only one patient complained of adverse reaction. So in the treatment of coexisting candidiasis and inflammatory dermatoses the combination of clotrimazole 1% + beclomethasone 0.025% (candid-B cream) was found to be highly effective.

[A case of Lyme disease with the triad of neurologic manifestations (meningitis, radiculoneuritis, facial nerve palsy) and dermatitis of the nail roots].

A 23-year-old man experienced dysesthesia of the distal parts of four limbs and spilling of drinking water from the right corner of his mouth. He also experienced low grade fever, headache, and nausea. Neurological examination showed weakness of the right frontal, orbicularis oculi and orbicularis oris muscles, atrophy and weakness of the interosseous muscles in both upper limbs and dysesthesia in the distal parts of four limbs. Laségue's sign was also positive on the left side. Examination of CSF showed lymphocytic pleocytosis and an increase of total protein. Serum antibody against Borrelia garinii was positive as determined by ELISA. The patient was diagnosed as a case of Lyme disease. His symptoms were not alleviated by administration of ceftriaxone (2 g/day) for 15 days, and dermatitis of nail roots appeared. Following administration of penicillin G at a high dose (12 million units/day) for 20 days, both neurologic symptoms and dermatitis were alleviated. Since B. garinii was reported to move preferentially toward the sites of low temperature, dermatitis of nail roots may be one of the characteristic features of Lyme disease caused by B. garinii.