Revue mars

Survey on the fungal flora of the cloaca of healthy pet reptiles S. NARDONI, R. PAPINI, G.M. MARCUCCI, F. MANCIANTI Dipartimento di Patologia Animale, Profilassi ed Igiene degli Alimenti, Università di Pisa, Viale delle Piagge 2, 56124 Pisa, ITALY.

*Corresponding author: E-mail: [email protected]

SUMMARY

RÉSUMÉ

The aim of this survey was to identify the fungal flora colonizing the cloaca of healthy reptiles kept in a pet shop to be sold as pets. The examined reptiles were belonging to the sub-orders Sauria (n=140) and Ophidia (n=17) or to the order Testudines (n=61). Following culture, the samples showed the presence of fungi in 75.2% of them. Fourteen genera of filamentous fungi, including 8 Aspergillus species, were identified as well as 6 genera of yeasts, including 13 species of Candida, 1 of Cryptococcus, and 1 of Pichia. A great number of the fungi isolated were known to be opportunistic agents of mycosis both in reptiles and humans. Therefore, the possible role of pet reptiles as carriers of opportunistic fungi into the human environment should be carefully evaluated.

Enquête sur la flore fongique du cloaque de reptiles sains de compagnie

Keywords: Pet reptiles, fungal flora, opportunistic fungi, zoonosis.

Le but de ce travail était d’identifier la flore fongique du cloaque de reptiles sains, présents dans une animalerie pour être vendus comme des animaux de compagnie. Les reptiles examinés appartenaient aux sous-ordres Sauria (n=140) et Ophidia (n=17) ou à l’ordre Testudines (n=61). Après culture, les échantillons ont montré la présence de champignons chez 75,2% d’entre eux. Quatorze genres de champignons filamenteux, dont 8 espèces d’Aspergillus, ont été identifiés ainsi que 6 genres de levures, dont 13 espèces de Candida, 1 de Cryptococcus et 1 de Pichia. Un grand nombre des champignons isolés est connu pour être des agents opportunistes de mycose chez les reptiles et chez l’homme. Le possible rôle des reptiles de compagnie en tant que porteurs de champignons potentiellement pathogènes dans l’environnement humain doit donc être attentivement évalué.

Mots-clés : Reptiles de compagnie, flore fongique, champignons opportunistes, zoonose

Introduction The number of reptiles kept as exotic pets in households was dramatically increased during the last years. Consequently, the risk of acquiring infections transmitted by these animals is also progressively increased. Although Salmonella species represent with no doubt the most important pathogenic agents transmissible from infected reptiles to man, it is also known that reptilian species can carry other potential zoonotic agents, including opportunistic fungi [8]. As pets, reptiles can live in close vicinity of man and, thus, they are likely to play an important role in spreading potential fungal pathogens in households and in their transmission to humans. Indeed, spores of pathogenic fungi excreted from reptiles via faeces and urine could increase their prevalence in the environment shared with humans. This could raise the risk of exposure to these fungi, leading to opportunistic fungal infections [3]. Opportunistic fungal infections have become increasingly common and, concomitantly, there has been a gradual rise in the number of rare fungal infections both in immunocompromised and normal patients [12]. A case of mycotic brain abscess caused by a Chrysosporium anamorph of Nanniziopsis wriesii, a known fungal pathogen in reptiles, Revue Méd. Vét., 2008, 159, 3, 159-165

has been reported in a HIV-seropositive man [19]. Consequently, the epidemiological role that reptiles, living in close vicinity of man, may play as carriers of opportunistic fungi needs to be accurately assessed. Many studies have been carried out on the presence of fungi in internal organs of free living lizards (Agama agama), wall geckos (Hemidactylus sp.) and turtles (Chelonia mydas) [5], the gut of caught garden lizards (A. agama) [3], various body sites of captive Chelonia and Squamata with a variety of unspecified clinical signs [11], cloacae of free ranging sea turtles [16], shell lesions of captive soil turtles (Testudo graeca, Testudo hermanni and Testudo horsfieldii) [4], skin samples of captive healthy Squamata [15], and faecal samples of wild eastern box turtles (Terrapene carolina carolina) [9]. However, reports of fungi isolated from the cloacal tract of pet reptiles are scant. The available data are limited to yeasts in diseased and necropsied animals [11] or to what can be extrapolated from wild animals [3, 5, 9, 16] and from other body sites of sampling [4, 15]. In contrast to previous investigations therefore the present study surveyed the presence of fungi in the cloacal content of healthy reptiles kept in a pet shop to be sold as companion animals.

NARDONI (S.) AND COLLABORATORS

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Materials and methods ANIMALS AND SPECIMEN COLLECTION From December 2005 to July 2006, 21 species of apparently healthy reptiles belonging to the sub-orders Sauria (n=140) and Ophidia (n=17) or to the Order Testudines (n=61) were investigated. Details are given in Table I. All the animals (n = 218) were captive-bred. At the time of sampling, they were living in terrariums or aquariums reproducing different types of ecosystems and were housed in a pet shop located in the city of Florence (Italy), where they were kept for sale. In order to document the isolation of fungi and their prevalence, cloacal samples were collected by gently inserting one sterile cotton swab into the cloaca of each animal and rotating the swab against the inner cloacal wall until faecal material was obtained. Immediately after collection, cotton swabs were dipped in 0.5 ml of sterile saline solution with added 5% gentamicin to avoid bacterial contamination, and kept at +4°C prior to analysis. Within 12 hours, the samples were transported to the laboratory to be processed as soon as possible.

CULTURE OF SAMPLES Samples were seeded on Petri dishes containing Malt Extract Agar (MEA, Difco Laboratories, Detroit, USA) as solid culture medium to allow isolation of moulds, and on MEA with added 10% biphenyl to encourage selective growth of yeasts. After seeding, all the plates were incubated at 25°C. Following seeding, from day 4 to day 7, the agar plates were inspected daily for fungal growth. When fungal colonies were detectable, subcultures were carried out to obtain pure cultures suitable for identification.

IDENTIFICATION OF ISOLATED FUNGI Filamentous fungi were identified on the basis of macroscopic and microscopic morphological characteristics. Typing was achieved to the genus level except for fungi belonging to the genus Aspergillus, where the identification was conducted to species level according to the keys of RAPER and FENNEL [17]. Yeasts were identified on the basis of morphological, physiological, and biochemical characteristics, including “germ tube test”, urease production, and carbohydrate assimilation using the API ID 32C® system (BioMérieux, Roma, Italia). Identifications were achieved to the genus level except for yeasts belonging to the genus Candida, Cryptococcus, and Pichia where the identification was conducted to species level. The final classification was made according to BARNETT et al. [1].

STATISTICAL ANALYSIS Animals were categorised according to habitat (aquatic or terrestrial), feeding (carnivorous, omnivorous or vegetarian), and zoological classification (Testudines, Sauria or Ophidia). Positivity rates and prevalence of fungi were determined as the number of positive samples/total number of samples x 100. The corresponding 95% confidence intervals (95% CI)

were also calculated. Differences between groups were compared by the Chi-square test, where a P value ≤ 0.05 was considered significant. Descriptive statistics and comparison of results were performed with Excel 6.0 for Windows 95.

Results Overall, fungi were found in 164 out of 218 (75.2±5.7%) cloacal samples. More specifically, fungi were isolated from 50 out of 61 (81.9±9.9%) Testudines, 103 out of 140 (73.5±5.6%) Sauria, and 11 out of 17 (64.7±22.8%) Ophidia. Omnivores had the highest prevalence rate (39/48, 81.2±11.1%), vegetarians had an intermediate rate (102/135, 75.5±7.3%), and carnivores had the lowest one (23/35, 65.7±15.8%). Thirty-six out of 43 (83.7±11.2%) aquatic reptiles and 128 out of 175 (73.1±6.6%) terrestrial reptiles tested positive. Despite these trends, no statistically significant difference was detected. With the exception of the black tailed rattlesnake (Crotalus molossus), cloacal isolated fungi were detected in representatives of all the other reptiles sampled, as shown in Table I. Different distributions of fungal occurrence were found in positive animals: moulds with one (n=46), two (n=27), three (n=11), or four (n=2) types of isolated agents, yeasts with one (n=31) or two (n=4) types, and various combinations of them (n=38). The occurrence of multiple (from 2 to 6) types of isolated fungi within a single host was common, being present in 82 of the positive samples (50±10.8%). On the other hand, many isolated fungi could be obtained from different hosts. Altogether, 14 genera of moulds, including 8 Aspergillus species, were identified as well as 6 genera of yeasts, including 13 species of Candida, 1 of Cryptococcus, and 1 of Pichia. The occurrence of moulds and yeasts in the sampled reptiles are shown in Table II and III, respectively. Penicillium sp. (39.9±6.5%) was the most commonly occurring isolated organism followed by Acremonium sp. (18.3±5.1%). Cladosporium sp. and Aspergillus niger were found in 8.2±3.6% and 6.4±3.1% of samples respectively, while Candida tropicalis, Geotrichum sp. and Trichosporon sp. were recovered from 5.5±2.9% of samples. Aspergillus versicolor and Mucor sp. occurred in 3.2±2.2% of samples. Candida krusei and Candida pelliculosa were isolated from 2.7±1.8% and 2.3±1.8% of samples respectively, while Paecylomyces sp. and Cryptococcus albidus were recovered from 1.8±1.3% of samples. Other isolated organisms, including Candida catenulata, Candida glabrata, Rhizopus sp. and Scopulariopsis sp. (1.4±1.3% each) as well as Alternaria sp., Aspergillus flavus, Aspergillus fumigatus, Aspergillus nidulans, Aspergillus ochraceous, Candida albicans, Candida famata, Candida guilliermondii, Candida norvegica, Fusarium sp., and Trichoderma sp. (0.9±1.2% each) were infrequently detected. The less common isolated organisms were Aspergillus oryzae, Aspergillus wentii, Candida lusitaniae, Candida membranaefaciens, Candida parapsilosis, Candida rugosa, Exophiala sp., Pichia etchelsii, Syncephalastrum sp., Verticillium sp. and Saccharomyces sp. (0.4±0.8% each). Due to heavy bacterial contamination, 9 (4.1%) of the isolated yeasts were lost in culture before they could be identified. Revue Méd. Vét., 2008, 159, 3, 159-165

FUNGI IN PET REPTILES

Reptile taxonomy L a t i n n a m e

Order Testudines

Order Squamata Suborder Sauria

Suborder Ophidia

161

Common name

Habitat

Regimen

Examined count

Positive count

Pseudemys scripta elegans

Red-Eared Turtle

Aquatic

Omnivorous

43

36

Geochelona carbonaria

Redfoot Tortoise

Terrestrial

Omnivorous

1

1

Testudo graeca

Greek Tortoise

Terrestrial

Vegetarian

1

1

Testudo hermanni

Hermann's Tortoise

Terrestrial

Vegetarian

13

9

Testudo marginata

Marginated Tortoise

Terrestrial

Vegetarian

2

2

Testudo pardalis

Leopard Tortoise

Terrestrial

Vegetarian

1

1

Chamaleo calyptratus

Veiled Chameleon

Terrestrial

Vegetarian

10

7

Eublepharis macularius

Leopard gecko

Terrestrial

Carnivorous

1

1

Gerrhosaurus validus

Giant African Plated Lizard

Terrestrial

Vegetarian

1

1

Iguana iguana

Green iguana

Terrestrial

Vegetarian

110

83

Paroedura pictus

Madagascar Ground Gecko

Terrestrial

Carnivorous

4

1

Physignathus cocincinus

Chinese Water Dragon

Terrestrial

Carnivorous

5

4

Pogona vitticeps

Bearded Dragon

Terrestrial

Carnivorous

5

4

Tiliqua scincoides

Eastern Blue-tongued Lizard Terrestrial

Omnivorous

2

1

Tupinambis merianae

Argentine Black and White Tegus

Terrestrial

Omnivorous

2

Boa costriptor

Boa Constrictor

Terrestrial

Carnivorous

1

1

Crotalus molossus

Black Tailed Rattlesnake

Terrestrial

Carnivorous

1

0

Elaphe guttata

Corn Snake

Terrestrial

Carnivorous

4

2

Lampropeltis triangulum

Milk Snake

Terrestrial

Carnivorous

1

1

Python molurus

Indian Python

Terrestrial

Carnivorous

3

2

Python regius

Royal Python

Terrestrial

Carnivorous

7

5

TABLE 1: Zoological taxonomy, habitat, regimen, number of reptiles examined and number of reptiles positive for fungi in their cloacal content. Revue Méd. Vét., 2008, 159, 3, 159-165

1


162

Moulds isolated

Acremonium

E x a m i n e d reptiles (n=218) Testudines (n=61)

Sauria (n=140)

Red-Eared Turtle (25)

Chinese Water Dragon (1)

Positive p e rc e n t a g e Ophidia (n=17) 18.3%

Green iguana (14)

Alternaria

Green iguana (2)

0.9%

A. flavus

Green iguana (1)

0.9%

Veiled Chameleon (1)

A. fumigatus


A. niger



Boa Constrictor (1)

Hermann's Tortoise (3)

Giant African Plated Lizard (1)

Corn Snake (1)

Green iguana (4)

Indian Python (1)

0.9% 6.4


A. nidulans


Madagascar Ground Gecko (1)

0.9%

A. ochraceous

Leopard Tortoise (1)

Green iguana (1)

0.9%

A. oryzae

Marginated Tortoise (1)

A. versicolor


Bearded Dragon (1)


Green iguana (3)

A. wentii

0.4% 3.2%

Green iguana (1)

0.4%

Green iguana (5)

8.2%

Exophiala

Green iguana (1)

0.4%

Fusarium

Green iguana (2)

0.9%

Green iguana (1)

3.2%

Cladosporium

Leopard Tortoise (1) Red-Eared Turtle (12)

Mucor

Greek Tortoise (1) Hermann's Tortoise (2) Marginated Tortoise (1) Red-Eared Turtle (2)

Rhizopus

Bearded Dragon (1)

Boa Constrictor (1)

1.4%

Corn Snake (1)

Paecylomyces

Leopard gecko (1) Veiled Chameleon (3)

Penicillium

1.8%


Bearded Dragon (3)

Corn Snake (1)

Redfoot Tortoise (1)


Real Python (4)

Greek Tortoise (1)

Green iguana (36)


Leopard gecko (1)



39.9%

Green iguana (3)

Scopulariopsis

Green iguana (3)

Syncephalastrum

1.4% Indian Python (1)

Trichoderma


Verticillium


Green iguana (1)

0.4% 0.9% 0.4%

TABLE 2: List of moulds isolated from the cloacal content of reptiles, animal species in which they were found (number of positive individuals per species in brackets), and percent of positive animals per mould isolated (/total number of examined reptiles). Revue Méd. Vét., 2008, 159, 3, 159-165


Moulds isolated

C. albicans C. catenulata

163

E x a m i n e d reptiles (n=218) Testudines (n=61) Hermann's Tortoise (2)

Sauria (n=140)

C. guilliermondii C. krusei C. lusitaniae C. membranaefaciens C. norvegica C. parapsilosis C. pelliculosa C. rugosa C. tropicalis Cr. albidus

Geotrichum Pichia etchelsii Trichosp.oron

Saccharomyces Unindentified

Ophidia (n=17) 0.9%

Chinese Water Dragon (1) Green iguana (1) Veiled Chameleon (1)

C. famata C. glabrata

Positive p e rc e n t a g e

1.4% Milk Snake (1) Royal Python (1)



Chinese Water Dragon (1) Green iguana (1) Green iguana (2) Argentine Black and White Tegus (1) Green iguana (3) Green iguana (1)

0.9% 2.7% 0.4% 0.4% 0.9%


Marginated Tortoise (2) Hermann's Tortoise (1) Hermann's Tortoise (1) Marginated Tortoise (1)

Red-Eared Turtle (4) Greek Tortoise (1) Hermann's Tortoise (1) Red-Eared Turtle (1) Red-Eared Turtle (1) Marginated Tortoise (1)

Madagascar Ground Gecko (1) Green iguana (1) Eastern Blue-tongued Lizard (1) Green iguana (3) Green iguana (1) Green iguana (11) Green iguana (2) Green iguana (12) Green iguana (1) Chinese Water Dragon (1) Green iguana (4)

Green iguana (6)

0.9% 1.4%

0.4% 2.3% 0.4% 5.5%

Royal Python (1)

Indian Python (1)

1.8% 5.5% 0.4% 5.5% 5.5% 0.4% 4.1%

TABLE 3: List of yeasts isolated from the cloacal content of reptiles, animal species in which they were found (number of positive individuals per species in brackets), and percent of positive animals per yeast isolated (/total number of examined reptiles).

Discussion All the isolated organisms found in the present survey are known to be widespread environmental contaminants with worldwide distribution. In addition, many of the yeast isolates have previously been reported as saprophytic commensals of reptiles [11]. Overall, despite some differences in prevalence rates, the range of isolated organisms obtained from the cloacae of pet reptiles in this survey is in agreement with findings of other investigations where the external or internal mycoflora of reptiles was examined [4, 11, 15]. According to FLAMANT et al. [4], since reptiles are poykilothermic animals, body temperature doesn’t play an important role in determining internal or external fungal flora. Differences in prevalence rates can probably be attributed to different sampling methods, study design, variety of animal species, number of animals examined, geographical location, and so on. It is not known when and how during captivity the positive reptiles acquired fungal contaminants in their cloacae. It is possible that reptiles were contaminated before they were Revue Méd. Vét., 2008, 159, 3, 159-165

carried into the shop. They probably acquired fungal contaminants in the cloacal content by ingestion of contaminated food (preys, vegetables) or water, by contact with faeces of other contaminated animals, or during defecation by contact with contaminated substrate which accumulated on the exterior of the cloaca. Many reptilian species shared the same types of isolated fungi. The basis of this sharing is uncertain but might be related to common exogenous sources of contamination. On the other hand, in many cases, different fungal isolates (up to six) could be obtained from a single positive swab. The distribution of the same fungal isolates in different hosts and the presence of many fungal isolates in a single host suggest that reptiles may act as facultative animal carriers for moulds and yeasts in their cloacae. All the animals sampled in this survey were symptom free, although 75.2% of them were harbouring fungi. This agrees with the results of other studies [3, 5, 15] and emphasizes the importance of histopathology in the diagnosis of fungal diseases in reptiles [7, 15]. It is known that many mycotic agents remain harmless in healthy individuals while they can become opportunistic in immunocompromised hosts. The opportu-

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nistic pathogenic role of most of the present fungal isolates has been well documented in reptiles and humans. With regard to the isolated moulds, the most prevalent genus was Penicillium sp. followed by Acremonium sp. and Cladosporium sp.. A Penicillium species has been reported as a cause of systemic mycosis in a Seychelles giant tortoise (Geochelone gigantea) and has been detected in association with A. niger and Curvularia lunata varaeria from superficial lesions on the skin and gingiva of saltwater crocodiles (Crocodylus porosus) and freshwater crocodiles (Crocodylus johnstoni) [7]. The genus includes thousands of species. Although Penicillium marneffei is by far the most important one, a number of diseases caused by other species have been reported in humans [12]. The genus Acremonium has been recovered from cases of mycetoma, onychomycoses, mycotic keratitis and as a colonizer of soft contact lens in man [18]. Cladosporium sp. has been reported along with Paecylomyces sp. from pulmonary nodules of maricultured green sea turtles (C. mydas) with mycotic pneumonia and in an adult anaconda (Eunectes murinus) with granulomatous lesions of the mandible [7]. It is one of the commonly reported agents of cutaneous phaeohyphomycosis in man [18]. Altogether, species of the genus Aspergillus were isolated with consistency. Aspergillus species have been identified in a St. Hilaire’s terrapin (Hydrastis hilarii) who died of generalized aspergillosis, from mycotic granulomas of the forefeet in a female musk turtle (Sternotherus odoratus), from pneumonic lesions of American alligators (Alligator mississippiensis), in association with Mucor sp. and Rhizopus sp. from cutaneous lesions of a 100-year-old American crocodile (Crocodylus acutus) with necrotizing dermatitis, in two San Esteban chuckwallas (Sauromalus varius) with edematous and necrotic lesions, in a black pointed teguixin (Tupinambis nigropunctatus) died for generalized mycosis, in a puff adder (Bitis arietans) with peritonitis [7], and from pulmonary lesions of two dead green anacondas (E. murinus) [13]. A. flavus, A. fumigatus, A. nidulans, A. niger, A. ochraceus, A. oryzae, and A. versicolor can cause allergic states, toxicosis, and opportunistic invasive infections in humans, though with different frequency. These fungi affect the respiratory tract in most of the cases but any organ can be involved [18].

Mucor sp., Paecylomyces sp., Rhizopus sp. and Scopulariopsis sp. were infrequently found in this survey. Mucor sp. and Rhizopus sp. have been reported in juvenile Florida softshell turtles (Apalone ferox) with necrotizing shell and skin lesions [7]. Mucor sp. has also been isolated from infected skin of wood turtles (Clemmys insculpta), pulmonary lesions of three species of crocodiles with fatal respiratory infections, and cutaneous lesions of a bearded dragon (Pogona barbata) [7]. In humans, Mucor sp. and Rhizopus sp. are amongst the aetiological agents of zygomycosis causing rhinofacial, rhinocerebral, cutaneous or subcutaneous forms and pulmonary, systemic, abdominal-pelvic or gastric disease [18]. A Paecylomyces species has been found to be associated with C. albicans in oral, gastric and hepathic nodular lesions in an Aldabra tortoise (G. gigantea) [7]. Paecylomyces species are uncommon human pathogens but have been documented in cases of mycotic keratitis, endocarditis and endophtalmitis [18]. A Scopulariopsis species has been recovered from shell


lesions of a captive soil turtle (T. graeca) [4]. In humans, Scopulariopsis sp. has been mostly associated with cutaneous and pulmonary pathologies [18]. The remaining isolated moulds, including Alternaria sp., Exophiala sp., Fusarium sp., Syncephalastrum sp., Trichoderma sp. and Verticillium sp. were only occasionally found. Alternaria sp. and Verticillium sp. have been identified in dermal lesions of two dead captive green anacondas (E. murinus) [13]. In humans, Alternaria sp. is frequently involved in cases of asthma and can colonize previously injured skin [18]. An Exophiala species has been reported as a cause of phaeohyphomycosis in a free living eastern box turtle (T. carolina carolina) [10]. Species of this genus are also amongst the aetiological agents of human phaeohyphomycosis [18]. Fusarium species have been identified as the cause of cutaneous mycosis in loggerhead sea turtles (Caretta caretta), as well as pathogens on crocodile farms, and as the cause of deep tissues mycosis in saltwater crocodiles (C. porosus) and freshwater crocodiles (C. johnstoni) [7]. Fusarium species are important agents of mycotic keratitis, onychomycosis and burned skin colonization in man [18]. Syncephalastrum sp. has been associated with mycotic dermatitis in two crocodiles [2] and can be an agent of human zygomycosis [18]. Trichoderma sp. has been involved in cases of necrotizing mycotic dermatitis in snakes [6] and can be a serious cause of peritonitis in human patients undergoing dialysis [18]. As far as yeasts are concerned, several Candida species were isolated in this survey. C. albicans has been identified as the causative agent of pneumonia in crocodiles and caimans, as well as recovered from multiple necrotic areas of the liver of a two-banded chameleon (Chamaeleo bitaeniatus), and from necrotic esophageal lesions in a crocodile tegu (Crocodilus lacertinus) [7]. Although Candida species are found in the normal flora of skin and digestive tract in man, C. albicans, C. glabrata, C. guilliermondii, C. krusei, C. lusitaniae, C. parapsilosis, and C. tropicalis are well known opportunistic human pathogens. Diseases caused by Candida species cover a range of pathologic effects, including oral, bronchial, pulmonary, alimentary, mucocutaneous and systemic candidiasis as well as vaginitis, balanopostitis, endocarditis and meningitis [18].

Geotrichum sp. and Trichosporon sp. were recovered less frequently. Geotrichosis has been documented in a group of captive carpet pythons (Morelia spilotes variegata) with mycotic dermatitis and in a northern water snake (Nerodia sipedon) with caseous subcutaneous nodules [7]. A Trichosporon species has been isolated from the liver and kidney of several captive banded rock rattlesnakes (Crotalus lepidus klauberi) [18]. In humans, lesions caused by Geothricum sp. are bronchopulmonary, bronchial, oral, vaginal, cutaneous, and very rarely alimentary, while Trichosporon sp. has been involved in a variety of opportunistic infections [18]. Finally, Cr. albidus, P. etchelsii and Saccharomyces sp. were only sporadically detected. Though Cryptococcus neoformans is the main aetiological agent of cryptococcosis in humans, Cr. albidus and a few other species have been isolated on rare occasions in severely immunocompromised Revue Méd. Vét., 2008, 159, 3, 159-165


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patients. Saccharomyces sp. has been implicated in several cases of pulmonary disease and mycosis of the stomach [18].

D., BOUCHARA J.P: Flore fongique des lésions de la carapace des tortues terrestres de compagnie dans l’ouest de la France. J. Mycol. Med., 2003, 13, 67-72.

Pet shops are common sources of new companion animals such as pet reptiles. Pet ownership is an important risk factor for the occurrence of many zoonoses, including mycoses. Living in close vicinity to man, reptiles could play an important role in the transmission of fungal agents to human beings [3]. The high isolation rate of fungi from cloacae of pet reptiles and the identification of opportunistic agents in the present survey suggest a potential human risk of opportunistic fungal infection from these pets. Extremely high dispersal of fungal spores has been found in faecal samples of reptiles [9]. Other authors have shown that reptiles from pet shops may harbour zoonotic infectious agents [14]. A case of mycotic brain abscess caused by a fungal pathogen of reptiles has been reported in a 38 year old, HIV-seropositive Nigerian man [19]. Therefore, our findings and those of other authors indicate that reptiles sold at pet shops in Italy may be considered as a potential source of human opportunistic mycoses. In addition, our results provide some insight about the normal cloacal mycoflora of healthy pet reptiles. At present, it is not known whether pet reptiles may be more active carriers of fungal agents in their cloacae, when compared to free living reptiles, or not. Such a difference might be related to environmental factors varying between free living and pet reptiles. For instance, the latter may be exposed to poor husbandry such as overcrowding, poor hygienic measures, high humidity, poor water and diet quality, and so on that might enhance the possibility of acquiring fungal contaminants. Since only one pet shop specialised in reptiles could be surveyed in the present study, further investigations are needed to clarify the true extent and public health implications of pet reptiles as carriers of opportunistic fungi.

5. - GUGNANI H.C., OKAFOR J.I.: Mycotic flora of the intestine and other internal organs of certain reptiles and amphibians with special reference to characterization of Basidiobolus isolates. Mykosen, 1979, 23, 260-268.

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