Coccidioides posadasii infection in bats, Brazil.
Mycoses (Health aspects)
HIV patients (Health aspects)
Cordeiro, Rossana de Aguiar
Silva, Kylvia Rocha de Castro
Brilhante, Raimunda Samia Nogueira
Moura, Francisco Bergson Pinheiro
Duarte, Nayle Francelino Holanda
Marques, Francisca Jakelyne de Farias
Cordeiro, Rebecca de Aguiar
Filho, Renato Evando Moreira
de Araujo, Roberto Wagner Bezerra
Bandeira, Tereza de Jesus Pinheiro Gomes
Rocha, Marcos Fabio Gadelha
Sidrim, Jose Julio Costa
|Publication:||Name: Emerging Infectious Diseases Publisher: U.S. National Center for Infectious Diseases Audience: Academic; Professional Format: Magazine/Journal Subject: Health Copyright: COPYRIGHT 2012 U.S. National Center for Infectious Diseases ISSN: 1080-6040|
|Issue:||Date: April, 2012 Source Volume: 18 Source Issue: 4|
Studies have demonstrated that bats (order Chiroptera) are
reservoirs for many infectious agents, including protozoa, bacteria,
viruses, and fungi (1). Several studies confirm that bats have a great
effect on human health because they can transmit numerous infectious
agents and provide a reservoir for emerging pathogens (1,2). The
interaction between these animals and pathogenic fungi is well
illustrated by the occurrence of histoplasmosis outbreaks in humans who
are exposed to bat droppings in the environment (3,4). In Brazil,
histoplasmosis is an endemic disease that occurs mainly in patients with
AIDS (5), but Histoplasma capsulatum var. capsulatum has also been
isolated from bats captured in urban areas (4).
To analyze the eco-epidemiologic aspects of H. capsulatum in northeast Brazil, we captured bats from urban and rural areas of Ceara State. However, the research revealed the existence of a bat that was naturally infected with Coccidioides posadasii and 2 other chiropterans with coccidioidal immunologic responses. This fungal pathogen can cause coccidioidomycosis, a serious infection in humans and animals. The mycosis is presently considered to be endemic to Northeast Brazil, as evidenced by human autochthonous cases (6-8), positive coccidioidin skin-test results (7), and isolation of the fungus from soil (7,9). We describe the isolation of C. posadasii in bats and discuss the epidemiologic effects of this finding.
From August 2010 to March 2011, a total of 83 bats of 7 species were captured in 6 cities in Ceara State, Northeast Brazil, where patients with histoplasmosis are seen: Ubajara, Itapiuna, Quixada, Russas, Aracoiaba, and Baturite. The animals were captured during the day (nonhematophagous bats) or night (hematophagous bats) by using nylon mist nets with 36-mm mesh. The study was part of the rabies control surveillance program headed by the Ceara State Health Department and was approved by the ethics committee of the State University of Ceara (process 07381395-8).
Immediately after capture, the bats were euthanized by an overdose of diethyl ether by inhalation, and their spleen, liver, and lungs were analyzed for H. capsulatum isolation. Fragments of each organ were homogenized by maceration in saline supplemented with 200 mg/L chloramphenicol. Aliquots of 100 [micro]L were seeded onto plates containing brain-heart infusion agar, supplemented with 1% glucose, 0.1% L-cysteine, 200 mg/L chloramphenicol, and 0.05% cycloheximide, and incubated at 25[degrees]C or 35[degrees]C for as long as 6 weeks (10). Remaining aliquots of each homogenate, as well as organ fragments, were kept at -20[degrees]C.
Although none of the samples were positive for H. capsulatum, a colony (Figure) suggestive of Coccidioides spp. (Figure, panel A) was isolated from lung homogenates (incubated at 35[degrees]C) from Carollia perspicillata bats. Microscopic analysis showed hyaline septate hyphae and arthroconidia alternating with empty disjunctor cells (Figure, panel B). Lung fragments from the infected bat were then removed from storage and examined by direct microscopy, revealing coccidioidal spherules (Figure, panel C). The suspected Coccicioides colony was evaluated through the in vivo reversion test (9). In brief, 5 mL of 0.9% saline was added to a well-sporulating slant culture (15 days old) that was then gently scraped with a cotton swab. Two mice were injected intraperitoneally with 1 mL of the homogeneous supernatant and then were held under biosafety level 3 conditions for 4 weeks. After this period, the animals were euthanized and their spleen, liver, and lungs were removed. Fragments were examined for coccidioidal spherules by direct microscopy with 10% potassium hydroxide and also cultured on BBL Mycosel Agar (Becton, Dickinson and Company, Franklin Lakes, NJ, USA). Additional histopathologic analyses of each organ were performed. Spherules with endospores were found in the lungs of the infected animals (Figure, panel D), and histopathologic analysis supported the identification of Coccidioides spp. (Figures 1, panels E, F). Fragments of the spleen, liver, and lungs cultured on Mycosel Agar yielded mold colonies that produced typical coccidioidal arthroconidia. An additional test was performed by specific PCR reaction (11) with Coi9-1F (5 '-TACGGTGTAATCCCGATACA-3') and Coi9-1R (5 '-GGTCTGAATGATCTGACGCA-3') primers to confirm the identity of the pathogen. A voucher of the fungal strain was deposited in the Specialized Medical Mycology Center Culture Collection at the Federal University of Ceara under code CEMM 05-5-059.
Homogenates of lungs, spleen, and liver of all bats were removed from storage and assayed by immunodiffusion tests specific for H.capsulatum and C. posadasii antigens (12) (ID Antigen H & M and IDCF Antigen; Immy Immunodiagnostics, Inc., Norman, OK, USA) according to the manufacturer's instructions. None of the homogenates showed positive reactions in H. capsulatum immunodiffusion tests. However, positive antibodies against Coccidioides spp. were found in 1 sample of lung from Glossophaga soricina bats. Positive antigen reactions were seen in homogenate liver samples from 2 animals, identified as G. soricina and Desmodus rotundus bats. These results suggest natural coccidioidal infection among the animals evaluated.
Positive C. perspicillata and G. soricina bats were captured in the same place, a deserted house in the urban area of Aracoiaba (4[degrees]21'59.1"S and 38[degrees]48'51.9"W) that has a semi-arid climate, with a rainy season from February through April and an average rainfall of 1,010.3 mm per year. The vampire bat, D. rotundus, was captured inside a cave in Ubajara (3[degrees]48'14.3"S and 40[degrees]52'46.2"W), a city characterized by a warm, subhumid tropical climate, with a rainy period from January through April and rainfall of 1,483.5 mm per year.
We thank the Secretaria de Saude do Estado do Ceara for technical support.
This work was supported by grants from the National Scientific and Technological Development Council (CNPq-process no. 306637/2010-3 and Programa de Capacitacao em Taxonomia 562296/2010-7).
Dr Cordeiro is professor of medical microbiology at Universidade Federal do Ceara, Brazil. Her research focuses on pathogenic fungi in humans and animals.
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Author affiliations: Universidade Federal do Ceara, Fortaleza-Ceara, Brazil (R.A. Cordeiro, K.R.C. Silva, R.S.N. Brilhante, F.J.F. Marques, R.A. Cordeiro, R.E. Moreira Filho, R.W.B. Araujo, T.J.P.G. Bandeira, M.F.G. Rocha, J.J.C. Sidrim); Instituto Federal de Educacao, Ciencia e Tecnologia, Ceara, Brazil (K.R.C. Silva) Universidade Estadual do Ceara, Fortaleza-Ceara (M.F.G. Rocha); and Secretaria da Saude do Estado do Ceara, Ceara (F.B.P. Moura, N.F.H. Duarte)
DOI: http://dx.doi.org/ 10.3201/eid1804.111641
Address for correspondence: Rossana Cordeiro, 10 50 Av. Domingos Olimpio, Apto. 501, Bairro Jose Bonifacio, CEP 60040-080, Fortaleza, Ceara, Brazil; email: email@example.com
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