Pneumocystis has been isolated from a wide range of unrelated mammalian hosts, including humans, domestic and wild animals. It has been demonstrated that the genome of Pneumocystis of one host differs markedly from that of other hosts. Also, variation in the chromosome and DNA sequence of Pneumocystis within a single host species has been observed. Since information about the occurrence and nature of infections in wild animals is still limited, the objective of this work was to detect the presence of Pneumocystis sp. in lungs of bats from two states from Brazil by Nested-PCR amplification. The bats, captured in caves and in urban areas, were obtained from the Program of Rabies Control of two States in Brazil, Mato Grosso and Rio Grande do Sul, located in the Mid-Western and Southern regions of the country, respectively. DNAs were extracted from 102 lung tissues and screened for Pneumocystis by nested PCR at the mtLSU rRNA gene and small subunit of mitochondrial ribosomal RNA (mtSSU rRNA). Gene amplification was performed using the mtLSU rRNA, the primer set pAZ102H -pAZ102E and pAZ102X -pAZY, and the mtSSU rRNA primer set pAZ102 10FRI -pAZ102 10R-RI and pAZ102 13RI -pAZ102 14RI. The most frequent bats were Tadarida brasiliensis (25), Desmodus rotundus (20), and Nyctinomops laticaudatus (19). Pneumocystis was more prevalent in the species Nyctinomops laticaudatus (26.3% = 5/19), Tadarida brasiliensis (24% = 6/25), and Desmodus rotundus (20% = 4/20). Besides these species, Pneumocystis also was detected in lungs from Molossus molossus (1/11, 9.1%), Artibeus fimbriatus (1/1, 100%), Sturnira lilium (1/3, 33.3%), Myotis levis (2/3, 66.7%) and Diphylla ecaudata (1/2, 50%). PCR products which could indicate the presence of Pneumocystis (21.56%) were identified in DNA samples obtained from 8 out of 16 classified species from both states (5 bats were not identified). This is the first report of detection of Pneumocystis in bats from Brazil.INDEX TERMS: Pneumocystis sp., bats, Nested-PCR.
do congelamento sobre a viabilidade de células leveduriformes. RESUMO O controle de produtos de origem animal está diretamente associado à sanidade animal e à saúde pública. Na suspeita de mastite, tanto bacteriana quanto micótica, o congelamento de amostras de leite permite aumentar o período de tempo para análise microbiológica, permitindo um correto diagnóstico laboratorial. As leveduras são os principais agentes ambientais envolvidos na mastite micótica. O objetivo do presente trabalho foi verificar se o congelamento afeta a recuperação de leveduras potencialmente presentes no leite. Foram utilizados seis isolados: Rhodotorula sp. (L1), Geotrichum sp. (L2), Cryptococcus sp. (L3), Candida tropicalis (L4) e Candida albicans (L5), oriundos de leite in natura, e um isolado de C. parapsilosis (ATCC 22019). Após a inoculação experimental, as amostras foram congeladas durante 10 dias, 3, 6 e 8 semanas. O congelamento do leite por períodos superiores a 10 dias causou redução no número de células viáveis. Sugerimos que sejam utilizados períodos mais curtos de congelamento para a análise micológica do leite.Descritores: mastite micótica, leite congelado, viabilidade celular, leveduras. ABSTRACTThe control of animal products is directly linked to the animal sanity and public health. When suspecting mastitis, either bacterial or mycotic, freezing milk samples increases the period of time for a microbiological analysis, thus allowing a correct laboratorial diagnosis. Yeasts are the main environmental agents involved in mycotic mastitis. The objective of the present work was to verify if freezing affects the recovery of yeasts present in the milk. Six isolates were used: Rhodotorula sp. (L1), Geotrichum sp. (L2), Cryptococcus sp. (L3), Candida tropicalis (L4) and Candida albicans (L5), isolated from milk in natura, and one C. parapsilosis (ATCC 22019) isolate. After experimental inoculation, the samples were frozen during 10 days, 3, 6 and 8 weeks. Milk freezing for periods longer than 10 days caused a reduction in the number of viable cells. We suggest that shorter freezing periods are used for the mycological analysis of milk.
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