Scanning electron microscopy of experimental adiaspiromycosis

Adiaspiromycotic granulomas of mice experimentally inoculated with fungusEmmonsia crescens Emmons et Jellison 1960 were examined by scanning electron microscopy. Their morphology, surface structures, and germinating adiaspores isolated from granulomas are described.     

Natural variability in the mycelial form of emmonsia crescens

A range of variations in colony characteristics and microscopical morphology of Emmonsia crescens Emmons & Jellison 1960 have been studied. A total of 13 strains of the mycelial stage of the fungus were examined in detail. The isolates originated from the lungs of 8 species of wild mammals collected in Czechoslovakia, Bulgaria, France and U.S.S.R. According to their colony appearance the strains were classified into 3 groups as follows: powdered, floccose and fluffy. The microscopic structure of the colonies was relatively unique, differences were found in the size of aleuries and the type of their walls. The morphological variability of E. crescens was not markedly influenced by host species or geographic origin of strains.     

Mutants ofEmmonsia crescens — Their pathogenicity and size of adiasporesin vivo

The pathogenicity of seven morphological mutants ofEmmonsia crescens was tested by means of intraperitoneal inoculation in mice. All mutants caused adiaspiromycosis. Adiaspores were isolated from granulomas after 2 months and their diameters were determined. Adiaspores from granulomas caused by five mutants (M-5, M-6, M-8, M-9 and M-16) were significantly smaller than adiaspores from granulomas caused by the wild strain, from which the mutants were derived. Two mutants (M-6 and M-9) produced adiaspores of the smallest diameter (130.5 and 119.9 μm) with the lowest variance of values, differing thus most from the original wild strain with adiaspores of 230.4 μm in diameter. A positive correlation was found between the size of the adiasporein vivo and growth rate of the mycelial stage ofEmmonsia crescens in vitro. The mutation characterized by the decreased growth rate of the mycelial stage is pheno-typically manifested in the adiasporic stage of the life cycle ofEmmonsia crescens, i.e. by the smaller average size of adiaspores in granulomas.     

Influence of animal hibernation on the development of mycoses

The development of adiaspiromycosis and trichophytosis depending upon the state of activity of redcheeked squirrels is described. The conidia of Chrysosporium (Emmonsia) parvum var. crescens, are not transformed into adiaspores when injected into hibernating animals. During the hibernation period of four months, most of the conidia die. After awakening, the remaining viable conidia are transformed into adiaspores. During hibernation the squirrels, which had been infected with Trichophyton mentagrophytes var. granulosum, developed a symptom-free infection.     

Adiaspiromycosis Due to <Emphasis Type="Italic">Emmonsia crescens</Emphasis> is Widespread in Native British Mammals

Adiaspiromycosis caused by Emmonsia crescens is primarily a respiratory disease affecting small mammals, especially members of the Families Rodentia, Carnivora and Mustelidae. Although isolated reports exist of adiaspiromycosis in free-living British wildlife, the extent of infection in wild animals in the UK, and the significance of any associated pathology are unclear. Here, we report the results of histopathological examination of lungs of free-living wild mammals from the south–west UK coupled with digestion of lung material in potassium hydroxide followed by centrifugation and microscopic examination for the presence of adiaspores. The combined results showed that almost one-third (27/94, 28.7%) of animals examined had evidence of infection with E. crescens. Attempts to culture E. crescens from infected lungs were unsuccessful. However, E. crescens could be confirmed as the causative agent by PCR amplification and sequencing of DNA from adiaspores micro-dissected from animal lungs. The prevalence of adiaspiromycosis was largely independent of animal species or precise geography. Adiaspore burdens in most animals were low, consistent with transient exposure to E. crescens. However, burdens in several animals suggested heavy or repeated exposures to E. crescens, and were considered sufficient to have significantly impaired respiratory function. Finally, since E. crescens is apparently widespread in UK mammals and the first UK human case of adiaspiromycosis was reported recently, we present data obtained using a previous isolate of E. crescens demonstrating that both the mycelial and adiaspore phases of the organism are susceptible to amphotericin B, voriconazole, itraconazole and caspofungin.     

In vitro formation of adiospores in 10 strains of Emmonsia cresens, the fungal agent of adiospiromycosis]

Emmonsia crescens Emmons and Jellison 1960 is a dimorphic fungus responsible for pulmonary mycoses in animals and man. Temperature was the essential factor influencing the development of the parasitic phase : the adiaspores, which formed better at 37 degrees than at 40 degrees. Certain media (medium with blood, with egg, Sabouraud...) also favoured the growth of adiaspores. The amount of CO2 in the ambiant atmosphere had no effect on the formation of adiaspore phase. On the other hand, the liquid medium Sabouraud was not suitable for certain strains wich were not able to develop adiaspores in semianaerobic media neither at 37 degrees nor at 40 degrees.     

Particularites de l'infestation adiaspiromycosique par Emmonsia crescens Emmons et Jellison, 1960 chez Pitymys subterraneus (De Selys-Longchamps 1836) et Mustela nivalis nivalis L. 1766

From the observation of some hundred lungs of small wild mammals in France, we have found two types of parasitism byEmmonsia crescens Emmons &Jellison, (1960). In most species, for example inApodemus sylvaticus andClethrionomys glareolus, the adiaspores ranged from 200 to 475 µ in diameter, their cell walls are 28 µ in average thickness and they cause a weak histological reaction in the parenchyma of the lungs. InPitymys subterraneus, the adiaspores are always much smaller, about 85 µ. They have a thin cell wall, 4 µ, and are surrounded by a strong histological reaction, the thickness of which ranges from 130 to 140 µ. The adiaspores observed inMustela nivalis nivalis are still smaller, about 70 µ, and they provoke a pulmonary reaction a hundred µ thick. However, we think that these small adiaspores belong to the speciesEmmonsia crescens Emmons &Jellison, (1960):

1. they germinate springing many mycelian tubes.
2. the macroscopic and microscopic morphological characteristics of the mycelian phase are identical to those of several strains ofEmmonsia crescens.
3. the mycelian phase completely turns into the yeast phase at 37° C on “Brain Heart Agar” Difco.
4. a strain isolated from aP. subterraneus, inoculated to a white mice, a bank vole or a field vole gives big adiaspores typical ofE. crescens in their respective lungs.
5. from an epidemiologic point of view, it is difficult to imagine such a narrow cohabitation of two species of so non specific germ having each its own host.

     

Seasonal incidence of adiaspores of Emmonsia crescens emmons & Jellison 1960 in wildly living animals

Summary In a collection of rodents from one locality a remarkable irregularity of monthly incidence of lung adiaspores was observed. The highest frequency of infection was ascertained in animals captured in April and May. The authors consider this phenomenon to be important for the elucidation of the life cycle ofEmmonsia crescens in nature.     

Adiaspiromycosis in large free living carnivores

Adiaspiromycosis was identified in 6 animals in the examination of the lungs of 90 large carnivores. Emmonsia crescens (Chrysosporium parvum var. crescens) was demonstrated as the causative agent in 5 cases of disease — in the badger (Meles meles), the otter (Lutra lutra) and the fox (Vulpes vulpes). E. parva was demonstrated in the remaining case of disease in a fox. The badger is a new, up to the present unknown host of E. crescens. The sporadic occurrence of adiaspiromycosis in the fox and the otter classifies this disease among rare diseases of these animals.     

The ultrastructure of the spherules of Emmonsia crescens studied by the technique of freeze-etching

Young spherules (adiaspores) of the fungusEmmonsia crescens from the cultures growing on Sabouraud's dextrose agar have been prepared for electron microscopy by the technique of freeze-etching. In the cytoplasmic membrane grooved invaginations corresponding with the ridges on the inner face of the cell membrane have been found. The surface of the cytoplasmic membrane is densely covered with regular globules, the diameter of which is about 100 Å. Mitochondria, endoplasmic reticulum, nuclear membrane with pores, and numerous stock materials in the form of fatty drops, and glycogen can be identified in the cytoplasm.

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Zusammenfassung Junge Sphärulen (Adiasporen) der auf Sabouraud's Glukoseagar wachsenden Kultur vonEmmonsia crescens wurden mit Hilfe der freeze-etching-Technik elektronenmikroskopisch studiert. An der cytoplasmatischen Membran wurden Invaginationen festgestellt. Diesen Invaginationen entsprechen Ausläufer der inneren Zellwandfläche. Die Oberfläche der cytoplasmatischen Membran ist mit regelmässigen Globulen (ca. 100 Å) dicht bedeckt. Im Cytoplasma konnten Mitochondrien, endoplasmatisches Retikulum, Kernmembran mit Poren sowie zahlreiche Reservestoff in Form der Fett-Tröpfchen und Glykogen identifiziert werden.

     

L'adiaspiromycose par Emmonsia crescens chez les petits mammifères sauvages en France

Résumé Dans le cadre d'un travail d'ensemble sur les petits mammifères sauvages en tant que réservoirs de germes pour un certain nombre d'anthropozoonoses, les auteurs ont recherché les adiaspores d'E. crescens dans les poumons de près de 3.000 pertits mammifères sauvages, en provenance de 45 départements français par la méthode d'homogénéisation-digestion précédemment décrite. Ces animaux appartiennent à 52 espèces de carnivores, rongeurs, insectivores et chiroptères.Les pourcentages moyens d'infection révèlent que l'adiaspiromycose des petits mammifères sauvages semble être une affection banale en France.Il existe cependant une variation dans l'intensité et la fréquence du parasitisme selon l'espèce, la famille, parfois l'ordre considérés. Ainsi, aucune des 109 chauves-souris examinées, appartenant à 17 espèces différentes, n'a été trouvée porteuse d'adiaspores. Les pourcentages d'animaux parasités varient, dans les autres ordres, de 0 à 50% selon les espèces.Le climat, la période de l'année, l'altitude ne semblent pas influer sur l'existence de cette parasitose.

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Within the framework of investigation on small wild mammals concerning the carrier stage of microorganisms in a certain number of anthropozoonoses, the authors investigated for adiaspores ofEmmonsia crescens the lungs of almost 3,000 small wild mammals from 45 departments of France by means of the method of homogenisation-digestion. The mammals belonged to 52 species of carnivors, rodents, insectivors, and chiropters. The mean percentage of infection revealed, that the adiaspiromycosis of small wild mammals is a banal affection in France. There is, however, a variation in intensity and frequency of parasitism according to species, family, sometimes according to order. None of the 109 bats, belonging to 17 different species, was found as carriers of adiaspores. The percentage of animals infected in the other orders varied between 0 to 50 per cent according to species. The climate the period of the year, the altitude do not appear to influence the existence of this parasitism.

     

Taxonomic study of keratinophilic fungi isolated from soil and some mammals in France

In France, most of the researches have showed the distribution of dermatophytes and other keratinophilic fungi in the soil and in the coat of wild mammals. During these last years, we have also practiced numerous investigations about the distribution of such fungi. The isolated species are: Trichophyton mentagrophytes, Trichophyton terrestre, Trichophyton ajelloï, Trichophyton simii, Microsporum gypseum, Microsporum cookei, Microsporum nanum, Microsporum persicolor, Anixiopsis stercoraria, Chrysosporium keratinophilum, Chrysosporium tropicum, Chrysosporium state of Arthroderma curreyi, Chrysosporium state of Arthroderma cuniculi, Chrysosporium state of Arthroderma tuberculatum, Chrysosporium state of Arthroderma multifidum, Ctenomycès serratus, Chrysosporium parvum, Chrysosporium indicum, Chrysosporium georgii, Chrysosporium merdarium, Chrysosporium asperatum, Chrysosporium pannorum.A key to these species is proposed: it attempts to provide an aid in identifying geophilic dermatophytes and related fungi (belonging to the Chrysosporium genus).     

The effect of fungizone on experimental adiaspiromycosis of laboratory mice

Summary In laboratory mice weighing about 25 g, with experimental infection of adiaspiromycosis, total inhibition of development of the parasitic phase of the agent of this mycosis is induced with Fungizone — Squibb in daily intraperitoneal administration of 0.03 mg (dissolved in 0.1 ml of isotonic saline). In autopsies carried out 10th, 20th and 30th day after an intraperitoneal inoculation of 1 ml suspension of the mycelial phase of two Czechoslovak isolates ofEmmonsia crescens Emmons &Jellison 1960, no elements of the parasitic phase — adiaspores were found in the organs of the treated mice. In control groups of the untreated mice, experimental infection developed which was demonstrated by macro- and microscopical controls of organs and by a successful recovery of the agent.     

Isolation of Emmonsia crescens Emmons et Jellison 1960 from small mammals in Bulgaria

Authors examined a total of 3849 specimens of mammals included in 36 species from different region of Bulgaria. Adiaspiromycosis was detected microscopically in 18 mammals species. Strains of Emmonsia crescens Emmons et Jellison 1960 was isolated from 6 species--Apodemus flavicollis, Apodemus sylvaticus, Clethrionomys glareolus, Microtus arvalis, Musmusculus and Rattus norvegicus.     

Experimental adiaspiromycosis of the common vole (Microtus arvalis) and other small wild mammals after intraperitoneal inoculation

After intraperitoneal inoculation ofE. crescens to nine species of wild small mammals, six species of rodents (Apodemus flavicollis, A. sylvaticus, Clethrionomys glareolus, Microtus agrestis, M. arvalis, Mus musculus) developed generalized adiaspiromycosis. The course of experimental infection corresponded to the infections of laboratory animals provoked in the same manner of inoculation. The authors studied the affliction of individual organs, the dynamics of growth of adiaspores in the organs of the abdominal cavity and in the lungs and followed up morphological changes in the adiaspores. In fresh cover glass preparations, the presence of manifestations was demonstrated justifying considerations on the capacity of multiplication of the adiaspiromycosis agent in the host organism.     

Some morphological observations concerning problems of endosporulation in Emmonsia crescens — Aetiological agent in adiaspiromycosis

Morphologic study dealing with problems of endosporulation in some spherules ofEmmonsia crescens.From the Departments of Pathology and Biology, Medical Faculty of Palacký University, Olomouc, Czechoslovakia.     

Human hair colonizing fungi in water sediments of India

Out of 144 samples of water sediments, 183 isolates belonging to 9 genera and 22 species were isolated. Fifty-nine isolates of Acremonium, 26 of Chrysosporium indicum, 22 of Chrysosporium keratinophilum, 17 of Malbranchea sp. and 10 of Microsporum gypseum were recovered. Acremonium implicatum, Chrysosporium georgii, Chrysosporium xerophilum and Geomyces pannorum were reported for the first time from India. This revised version was published online in June 2006 with corrections to the Cover Date.     

UV-sensitivity and repair of UV-damages in Salmonella of wild type.

Summary The UV-sensitivity of wild type Salmonella strains has been compared to that of wild type E. coli and its UV-sensitive mutants. Many wild type Salmonella strains are 4–5 times more sensitive than wild type E. coli and their inactivation curve is similar to that for E. coli with a mutation in the polA gene. Alkaline sucrose gradient centrifugation has shown a deficiency of these strains in normal excision repair of UV-damaged DNA. This deficiency is not a Salmonella genus feature because one strain as resistant as wild type E. coli was found. This resistant strain showed normal excision repair in alkaline sucrose gradient centrifugation experiments. The possible influence of plasmids and mutations in repair genes on the ability of Salmonella to repair UV-damaged DNA is discussed.     

Emmonsia parva (Emmons et Ashburn, 1942) Ciferri et Montemartini, 1959 = Chrysosporium parvum (Emmons et Ashburn) comb. nov.; Emmonsia crescens Emmons et Jellison, 1960 = Chrysosporium parvum var. crescens

Résumé L'étude comparative au cours de différents travaux antérieurs de la morphologie saprophytique et parasitaire in vitro et in vivo de vingt deux souches d'Emmonsia nous fait penser que ce genre est bien synonyme du genreChrysosporium et de l'espèceC. parvum. Elle nous permet surtout de conclure que l'on peut différencier deux ou trois variétés selon que l'on privilégie le comportement parasitaire in vitro ou in vivo.La priorité donnée au comportement in vitro donc à la thermophilie des souches nous permet de distinguer comme Carmichael (1962) deux variétés:Chrysosporium parvum etC. parvum var. crescens dont nous précisons les caractères distinctifs.

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Thanks to comparative studies of the saprophytic and parasitic morphologies of 22 strains ofEmmonsia in vitro and in vivo, we think that it is synonymous with the genusChrysosporium and the speciesC. parvum. In this species, we can distinguish 3 varieties if you consider the one obtained in vivo.The priority given to the thermophily of the strains permits us to distinguish, as Carmichael did (1962), 2 varieties:Chrysosporium parvum andC. parvum var. crescens of which we state precisely the distinctive characteristics.

     

Keratinophilic fungi isolated from the air at Pavia

The presence of keratinophilic fungi was revealed by sampling the air of Pavia (Italy) from March 1981 to February 1982. The species isolated were: Chrysosporium indicum, Geomyces pannorum var.pannorum, Microsporum gypseum, Myceliophtora vellerea and Trichophyton terrestre. Several filamentous fungi tolerating high levels of cycloheximide were also recovered.