Two new lipid-dependent Malassezia species from domestic animals

During a study on the occurrence of lipid-dependent Malassezia spp. in domestic animals, some atypical strains, phylogenetically related to Malassezia sympodialis Simmons et Guého, were shown to represent novel species. In this study, we describe two new taxa, Malassezia caprae sp. nov. (type strain MA383=CBS 10434), isolated mainly from goats, and Malassezia equina sp. nov. (type strain MA146=CBS 9969), isolated mainly from horses, including their morphological and physiological characteristics. The validation of these new taxa is further supported by analysis of the D1/D2 regions of the 26S rRNA gene, the ITS1-5.8S-ITS2 rRNA, the RNA polymerase subunit 1 and chitin synthase nucleotide sequences, and the amplified fragment length polymorphism patterns, which were all consistent in separating these new species from the other species of the genus, and those of the M. sympodialis species cluster, specifically.     

First description of sarcoptic mange in wild European rabbit (Oryctolagus cuniculus)

The European wild rabbit (Oryctolagus cuniculus) is a keystone species in the Iberian Mediterranean ecosystems being the staple prey of protected and endangered predators. Wild rabbits were once widespread, but the introduction of two viral diseases, myxomatosis in the 1950s and the rabbit hemorrhagic disease in 1989, resulted into a dramatic decline of its populations. Sarcoptic mange is a highly contagious parasitic infection caused by Sarcoptes scabiei. The first cases of sarcoptic mange in a wild rabbit population are recorded from a hunting area in Majorca (Balearic Islands, Spain). Five of 32 inspected rabbits (14.7%) were affected, with similar prevalences in summer and autumn. Sarcoptes scabiei were obtained from the edge of the lesions of two of the rabbits. The most frequently observed lesions were small areas of alopecia and crusts in the limbs. Affected limbs presented also a marked hypertrichosis and an apparent abnormal growth of the nails. One rabbit also presented lesions around mouth and nostrils. Parasitized rabbits were in significant lower body condition than healthy ones. According to previous reports of mange epizooties in other naïve wild species that caused marked short-term effects in their populations, this disease may pose a risk for the conservation of wild rabbit and their predators. Whether mange is endemic in rabbits from Majorca or it has been introduced is unknown. Game managers are encouraged to be aware of introducing sarcoptic mange during rabbit translocations.

     

马拉色菌相关人群及正常人群耳耵聍中马拉色菌带菌情况的调查

目的调查马拉色菌相关人群及正常人群耳耵聍中马拉色菌带菌情况。方法用结晶紫染色法对96例被调查人群耳耵聍进行马拉色菌检测,同时作培养,并以标准株作对照,用生理生化方法将耵聍中分离到的79株马拉色菌进行分类。结果马拉色菌相关人群耳耵聍中马拉色菌的直接检出率为91．84％（45／49）,培养阳性率为81．63％（40／49）,其中厚皮马拉色菌8株（16．33％）,合轴马拉色菌10株（20．41％）,糠秕马拉色菌22株（44．90％）。正常人群耳耵聍马拉色菌直接检出率为89．36％（42／47）,培养阳性率为78．72％（37／47）,其中厚皮马拉色菌5株（10．64％）,合轴马拉色菌8株（17．02％）,糠秕马拉色菌23株（48．93％）,斯洛菲马拉色菌1株（2．13％）。结论马拉色菌为正常人群及马拉色菌相关人群外耳道正常菌群,两组人群中马拉色菌的分离率和菌种分布无显著性差异。     

Onychomycosis due to Malassezia.

The records of fifty patients presenting Malassezia spp. associated onychomycosis were compiled from two different mycology laboratories from Medellín, Colombia. Malassezia spp. was isolated by culture as the only etiological agent in 32% of the cases and associated to a yeast of the genus Candida in 30% of the cases. In 22% of the cases although Malassezia spp. was observed by direct examination, it was no isolated but others species were obtained. No etiological agent was isolated by culture in 16% of the cases. We found evidence of the Malassezia spp.- Candida relationship in 48% of the cases by either direct examination or by culture isolation. The level of detection of Malassezia spp. by culture isolation was of 62% as compared to the direct examination. Results showed similar patterns of distribution of epidemilogical factors for both entities: onychomycosis by Candida albicans and onychomycosis by Malassezia spp.     

Microbe-I: fungal biota analyses of the Japanese experimental module KIBO of the International Space Station before launch and after being in orbit for about 460 days

In addition to the crew, microbes also find their way aboard the International Space Station (ISS). Therefore, microbial monitoring is necessary for the health and safety of the crew and for general maintenance of the facilities of this station. Samples were collected from three sites in the Japanese experimental module KIBO on the ISS (air diffuser, handrail, and surfaces) for analysis of fungal biota approximately 1 year after this module had docked with the ISS. Samples taken from KIBO before launch and from our laboratory were used as controls. In the case of KIBO, both microbe detection sheet (MDS) and swab culture tests of orbital samples were negative. The MDS were also examined by field emission-scanning electron microscopy; no microbial structures were detected. However, fungal DNAs were detected by real-time PCR and analyzed by the clone library method; Alternaria sp. and Malassezia spp. were the dominant species before launch and in space, respectively. The dominant species found in specimens from the air conditioner diffuser, lab bench, door push panel, and facility surfaces on our laboratory (ground controls) were Inonotus sp., Cladosporium sp., Malassezia spp., and Pezicula sp., respectively. The fungi in the KIBO were probably derived from contamination due to humans, while those in our laboratory came from the environment (e.g., the soil). In conclusion, the cleanliness in KIBO was equivalent to that in a clean room environment on the ground.     

正常人耵聍中马拉色菌菌种构成研究

目的研究正常人耵聍中马拉色菌菌种构成及同一宿主耵聍中菌种是否一致。方法采集45名健康志愿者双侧耵聍,0.1%曲拉通X-100溶解稀释后接种于含菜籽油培养基,生化及形态学方法鉴定到种,同时提取菌种DNA,用真菌通用引物ITS1/ITS4做PCR扩增并测序鉴定。结果有44例(97.78%)双侧耵聍中均培养出马拉色菌(共分离出88株菌),菌种构成:糠粃马拉色菌29株(32.95%)、斯洛菲马拉色菌23株(26.14%)、合轴马拉色菌18株(20.45%)、球形马拉色菌11株(12.50%)、限制性马拉色菌7株(7.95%)。44例(88株菌)中双侧耵聍菌种相同者有38例(76株菌)(一致率86.36%)。结论正常人耵聍中马拉色菌菌种分布较广,主要菌种为糠秕马拉色菌。同一宿主双侧耵聍中马拉色菌菌种具有一定的一致性。     

Occurrence of Malassezia species in healthy and dermatologically diseased dogs

The presence of Malassezia spp. yeasts was investigated in dermatological specimens of 224 dogs, 164 dermatologically diseased and 60 normal dogs. Subjects included in the study were of different breed, age, sex and habitat. Malassezia spp. positive cultures were obtained in 142 (63.4%) specimens: 67.6% from dermatologically diseased subjects and 51.6% from healthy dogs. Malassezia pachydermatis, either as a pure culture or in association with lipid-dependent species, was identified in 138 (97%) specimens. Malassezia furfur was identified in 69 (48.6%) specimens and was associated with other Malassezia species in 68 dogs, as a pure culture in one subject: at the best of our knowledge, this species was identified before as the sole species from canine dermatitis. Malassezia sympodialis was identified in 11 (7.7%) specimens, always in association with other species: it was never isolated from kennel dogs. Statistical analysis of data showed a very significant difference (P < 0.01) in the prevalence of isolation of Malassezia spp. between animals with and without dermatological signs, and in the distribution of cultural burden between diseased and healthy dogs. A statistically significant difference (P<0.05) was also detected in the group of animals between 1- and 5-years of age. No significant difference was found between male and female dogs.     

In vitro modulation of human keratinocyte pro- and anti-inflammatory cytokine production by the capsule of Malassezia species

Malassezia spp. are commensal, cutaneous fungi that are implicated in seborrhoeic dermatitis. We hypothesize that the lipid-rich capsule of Malassezia spp. masks the organism from host detection, and depletion of this layer elicits an inflammatory response. To test this, preparations of capsulated or acapsular [10% (v/v) Triton X-100 treated], viable and nonviable, exponential or stationary phase Malassezia furfur, Malassezia globosa, Malassezia obtusa, Malassezia restricta, Malassezia slooffiae and Malassezia sympodialis, were incubated with normal human keratinocytes. Proinflammatory (IL-6, IL-8, IL-1alpha and tumour necrosis factor-alpha) and anti-inflammatory cytokine (IL-10) release and intracellular IL-10 concentrations were quantified using enzyme-linked immunosorbent assays. Capsulated Malassezia yeasts stimulated limited or no production of inflammatory cytokines, and increased intracellular IL-10 (P < 0.05). Removal of the capsule of many Malassezia preparations caused a significantly increased production of IL-6, IL-8 and IL-1alpha, and a decrease in intracellular IL-10. Notably, acapsular viable, stationary phase M. globosa caused a 66-fold increase in IL-8 production (P < 0.001) and acapsular nonviable, stationary phase M. furfur caused a 38-fold increase in IL-6 production (P < 0.001) and a 12-fold decrease in intracellular IL-10 (P < 0.001). These results support the hypothesis that the lipid layer of Malassezia spp. modulates cytokine production by keratinocytes. This has implications in the pathogenesis of seborrhoeic dermatitis.     

Parasitic zoonoses in Papua New Guinea

Relatively few species of zoonotic parasites have been recorded in humans in Papua New Guinea. A greater number of potentially zoonotic species, mostly nematodes, occur in animals but are yet to be reported from humans. Protozoa is the best represented group of those infecting man, with Giardia duodenalis, Cryptosporidium parvum, Cyclospora cayetanesis, Toxoplasma gondii, Sarcocystis spp., Entamoeba polecki, Balantidium coli and, possibly, Blastocystis hominis. The only zoonotic helminths infecting humans include the trematode Paragonimus westermani, the cestodes Hymenolepis nana, H. diminuta and the sparganum larva of Spirometra erinacea, and the nematodes Trichinella papuae and Angiostrongylus cantonensis and, possibly, Ascaris suum. Other groups represented are Acanthocephala (Macracanthorhynchus hirudinaceus)), insects (Chrysomya bezziana, Cimex sp., Ctenocephalides spp.), and mites (Leptotrombidium spp. and, possibly Sarcoptes scabiei, and Demodex sp.). One leech (Phytobdella lineata) may also be considered as being zoonotic. The paucity of zoonotic parasite species can be attributed to long historical isolation of the island of New Guinea and its people, and the absence until recent times of large placental mammals other than pig and dog. Some zoonotic helminths have entered the country with recent importation of domestic animals, in spite of quarantine regulations, and a few more (two cestodes, one nematode and one tick) are poised to enter from neighbouring countries, given the opportunity. Improvement in water supplies, human hygiene and sanitation would reduce the prevalence of many of these parasites, and thorough cooking of meat would lessen the risk of infection by some others.     

基于线粒体细胞色素氧化酶I（COI）序列的猪和兔四个疥螨分离株的系统发育关系分析

为了探讨兔疥螨分离株和猪疥螨分离株的分类地位, 采用PCR技术首次扩增了分离自中国猪和兔的4个疥螨分离株的线粒体细胞色素氧化酶I（COI）基因, 并与GenBank中注册的14个国外疥螨分离株的同源基因进行了比较。序列分析结果显示: 扩增的4个疥螨株COI基因长度均为1 427 bp, 序列间无插入、缺失, A+T含量（73%）明显高于G+C含量（27%）, 碱基组成存在明显偏移。猪和兔的4个疥螨分离株间的COI基因同源性较高（99.1%～100.0%）, 它们与澳大利亚人疥螨株、国外动物疥螨株的同源性范围为98.4%～99.6%。在构建的NJ树中, 分离自中国猪和兔的4个疥螨分离株同澳大利亚人疥螨分离株、国外动物疥螨分离株亲缘关系较近。根据疥螨COI基因同源性分析和系统树构建结果, 我们认为分离自中国猪和兔的4个疥螨分离株与澳大利亚人疥螨分离株以及国外的动物疥螨分离株均应属于同一个种。     

Epidemiology and variability of Malassezia spp

A short review on Malassezia spp., completed with our experience, is made. The main epidemiological characteristics with particular regard to the diffusion in several animal species, the characteristics of skin colonization (in particular of the dog) and the distribution of the different Malassezia spp. in some hosts are discussed. Lastly the main phenotypic and genotypic characteristics, referred to M. pachydermatis especially, were described, showing their high variability and differentiation.     

The pathogenesis of Malassezia yeasts

The genus Malassezia includes twelve species of yeast, many of which have been mainly associated with human and canine diseases. Malassezia pachydermatis colonizes the skin and mucosal sites of healthy dogs and cats. Despite being part of the normal cutaneous microflora, Malassezia spp. yeast may become pathogenic under certain circumstances. This article reviews the factors related to both host and yeast which affect the pathogenical or commensal phenotypes of Malassezia yeasts.     

Concordance between phenotypical features and PCR-REA for the identification of Malassezia spp

The genus Malassezia has been recently revised and nowadays includes 11 species that cannot always be differentiated from each other by physiological and morphological tests. This study was aimed to evaluate the correlation between a molecular method and conventional phenotypic features in the identification of Malassezia spp. To achieve this aim, 92 Argentinean clinical strains isolated between 2001 and 2005 were analyzed along with three reference strains (Malassezia furfur CBS 7019, Malassezia sympodialis CBS 7222 and Malassezia slooffiae CBS 7956). By using PCR and restriction enzyme analysis with three different DNA endonucleases (PCR-REA), the molecular method consistently identified all three reference strains and all 92 clinical isolates as follows: 63 M. sympodialis, 18 M. furfur, 10 Malassezia globosa and one Malassezia obtusa. Phenotypic studies undentified 85 clinical isolates and two of the reference strains (total agreement > 91%). In particular for M. sympodialis, M. furfur and M. globosa, the species more frequently involved in human pathology, the agreement ranged between 84 and 96%. This result suggests that phenotypic studies are suitable for the presumptive identification of important Malassezia species in the clinical medical mycology laboratories where molecular methodologies are not available.     

Recent developments in the immunology and biology of Malassezia species

Malassezia spp. are members of the normal cutaneous flora, but are also associated with several cutaneous diseases. Recent studies of the interaction of Malassezia spp. with melanocytes, fibroblasts, keratinocytes and dendritic cells have highlighted their potential to modulate the immune response directed against them. In normal skin they may downregulate the inflammatory response, allowing them to live as commensals. In contrast, in atopic/eczema dermatitis syndrome and psoriasis, they may elicit an inflammatory response that contributes to the maintenance of lesions. Future research may define ways to influence this inflammatory cycle and hence to control or prevent exacerbations of these diseases.     

A comparison of the blood parasites in three subspecies of the yellow wagtail Motacilla flava

One-hundred and eighty yellow wagtails Motacilla flava belonging to 3 subspecies (Motacilla flava feldegg, Motacilla flava flava, Motacilla flava thunbergi) were caught during the spring migration in south Kazakhstan and investigated by microscopic examination of stained blood smears. Haemoproteus anthi, Haemoproteus motacillae, Leucocytozoon fringillinarum, Leucocytozoon majoris, Plasmodium relictum, Plasmodium polare, Atoxoplasma sp., Trypanosoma sp., and microfilariae were identified. The overall prevalence of infection was 47.8%. Prevalences of Haemoproteus spp. (27.2%), Plasmodium spp. (25.0%), Leucocytozoon spp. (8.9%), Atoxoplasma spp. (4.4%), Trypanosoma spp. (1.1%), and microfilariae (0.6%) were recorded. No differences were discernible in parasite fauna or intensities of infection between males and females or between different subspecies. However, prevalence of infection of Haemoproteus, Leucocytozoon, and Plasmodium spp. was different in different host subspecies. These differences can be explained by differences in geographical location of breeding areas of these birds.     

Potential pathogens carried by Spanish ibex (Capra pyrenaica hispanica) in southern Spain

The Spanish ibex (Capra pyrenaica hispanica) population of southern Spain was surveyed for potential pathogens associated with the conjunctiva, external ear canal, as well as reproductive and upper respiratory tracts. We sampled 321 ibex (131 adult males, 100 adult females, and 90 yearlings); these included 271 apparently healthy animals and 50 that were naturally infected with Sarcoptes scabiei. A total of 688 bacterial isolates were identified (377 gram-negatives, 225 gram-positives, and 86 Mycoplasma spp.); sex, age, location, infection with S. scabiei, and disposition of the animal (free-ranging versus captive) were evaluated as risk factors for infection. Infections with Mycoplasma agalactiae and Mycoplasma arginini were associated with age, having a higher frequency of isolation in young animals. With Escherichia coli, Mannheimia haemolytica, Pasteurella multocida biotype A, and Staphylococcus aureus, significantly higher isolation rates were associated with adults. The isolation frequency for E. coli was higher in females, whereas Moraxella bovis isolations were mostly associated with males. The presence of mange increased the risk of infection with both Streptococcus equi subsp. zooepidemicus and M. haemolytica. The geographic origin of sampled animals was related to the isolation of Branhamella ovis, M. agalactiae, and all Pasteurella sp. Isolations of M. haemolytica, P. multocida biotype A, E. coli, and B. ovis were more prevalent in samples from free-ranging rather than captive animals. Of the gram-positive bacteria, S. aureus represented the predominant species isolated from nasal, vaginal, and ocular samples. Mycoplasma agalactiae and M. arginini were the predominant Mycoplasma spp., and both were associated most often with the external ear canal. The most frequently isolated gram-negative bacteria included E. coli, M. haemolytica, P. multocida biotype A, and B. ovis. Isolation rates of gram-negative species varied by source. In nasal samples, M. haemolytica and P. multocida biotype A were isolated most frequently, whereas in ocular and vaginal samples, B. ovis and E. coli, respectively, were most frequently isolated.     

Conservation of Malassezia strains in blotting paper

Reference strains belonging to the genus Malassezia were analyzed to evaluate, by comparison, different preservation systems such us subculture, freezing at -80 degrees C in glycerol, and blotting paper-disc conservation. The viability, phenotypic and genotypic characteristics of the strains used in this study was evaluated. The blotting paper method was found to be advantageous to preserve Malassezia spp strains due to both, its simple implementation in the laboratory and its efficiency.     

The effects of a pesticide program on microbial populations from apple leaf litter

The leaf litter microbial community was quantitatively and qualitatively changed when a standard pesticide schedule that comprised an insecticide, a bactericide, and a fungicide was applied to McIntosh apple trees in the summer. Effects were observed for two winters by four indirect assays and three direct methods. Populations were altered qualitatively both years, but the most striking difference was the quantitative impact from year to year. Bacteria, filamentous fungi, and yeasts from treated leaves were reduced 10- to 10 000-fold between November 1976 and April 1977 and did not recover until snow cover had melted in March. Reductions in 1977-1978 were negligible. The marked seasonal difference is attributed to meteorological influences. Fluorescent pseudomonads were among the bacteria depressed by chemicals. Of the 49 genera of fungi and yeasts isolated, Coniothyrium sp., Penicillium spp., Arthrobotrys spp., and Nodulisporium sp. were appreciably reduced, whereas Typhula spp., Pleurophomella sp., Sporobolomyces spp., and Rhodotorula spp. were substantially enhanced by the spray program.     

Hematozoa from montane forest birds in Papua New Guinea

Blood smears were examined from 141 montane forest birds of 45 species in southeastern Papua New Guinea. Haemoproteus spp. occurred in 46 (32.6%), Leucocytozoon fringillinarum Woodcock, 1910 in five, Trypanosoma sp. in one and Haemogregarina sp. in one. Intensity of infection by Haemoproteus was highest in those avian species and families with the highest prevalence; increasing altitude had no demonstrable effect on the prevalence of Haemoproteus spp.     

Analysis of Malassezia microbiota in healthy superficial human skin and in psoriatic lesions by multiplex real-time PCR

Yeasts from the genus Malassezia are members of the normal biota of human skin, and may play a role in dermatopathology. Our previous study of the fungal microbiota from healthy subjects and from patients with psoriasis using clone library analysis revealed the presence of five Malassezia species and four uncharacterized phylotypes. We now compared the Malassezia microbiota from six healthy body locations and two psoriatic lesions, and evaluated its stability over time using multiplex real-time PCR. Samples from each body location were obtained monthly, for 4 months. Dual-labeled probes were designed to recognize four Malassezia sp. and two uncharacterized groups, and a genus-specific probe was also developed. A good correspondence was obtained between real-time PCR data and clone library analyses. Malassezia restricta was the most abundant species in the majority of samples, and high amounts of Malassezia globosa were also detected. The uncharacterized phylotype 1 was usually detected in lower proportions, nevertheless it was present in most samples. The microbiota was host-specific and relatively stable over time. In accordance with our previous observations, no significant dichotomy between samples from healthy skin and from psoriatic lesions was found; the samples clustered according to the subject, rather than health status.