Detection of Pneumocystis in lungs of bats from Brazil by PCR amplification / Detecção de Pneumocystis em pulmões de morcegos no Brasil por Nested-PCR

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 percent = 5/19), Tadarida brasiliensis (24 percent = 6/25), and Desmodus rotundus (20 percent = 4/20). Besides these species, Pneumocystis also was detected in lungs from Molossus molossus (1/11, 9.1 percent), Artibeus fimbriatus (1/1, 100 percent), Sturnira lilium (1/3, 33.3 percent), Myotis levis (2/3, 66.7 percent)and Diphylla ecaudata (1/2, 50 percent). PCR products which could indicate the presence of Pneumocystis (21.56 percent) were identified in DNA samples obtained from 8 out of 16 classified species from both states (5 bats were not identified). This is the ...

Pneumocystis tem sido isolado de uma grande variedade de hospedeiros mamíferos, incluindo humanos, animais domésticos e selvagens. Tem se demonstrado que o genoma do Pneumocystis de um hospedeiro difere marcadamente do de outros, assim como há variação no cromossomo e na seqüência de DNA dentro de uma única espécie de hospedeiro. Sabendo que a informação da ocorrência e natureza da infecção em animais silvestres ainda é limitada, o objetivo do trabalho foi detectar, por Nested-PCR, a presença de Pneumocystis sp. em pulmões de diferentes espécies de morcegos de dois estados do Brasil. Estes mamíferos voadores foram capturados em cavernas, áreas florestadas, de campo e urbanas pelo Programa de Controle da Raiva do Mato Grosso (região Centro-Oeste) e do Instituto de Pesquisas Veterinárias Desidério Finamor (RS) e Instituto Sauver no Rio Grande do Sul (região Sul). Os DNAs foram extraídos de 102 pulmões e realizado Nested-PCR utilizando os primers pAZ102H-pAZ102E e pAZ102X/R1-pAZY/R1 para amplificação do gene mtLSU-rRNA, e pAZ102 10F-RI - pAZ102 10R-RI e pAZ102 13-RI - pAZ14-RI para amplificação do gene mtSSU-rRNA. As espécies mais freqüentes foram Tadarida brasiliensis (25), Desmodus rotundus (20) e Nyctinomops laticaudatus (19). Pneumocystis foi detectado com maior prevalência nas Nyctinomops laticaudatus (26,3 por cento = 5/19), Tadarida brasiliensis (24 por cento = 6/25) e Desmodus rotundus (20 por cento = 4/20). Além destas espécies, Pneumocystis foi também detectado nos pulmões de Molossus molossus (1/11, 9,1 por cento), Artibeus fimbriatus (1/1, 100 por cento), Sturnira lilium (1/3, 33 por cento), Myotis levis (2/3, 66,7 por cento)e Diphylla ecaudata (1/2, 50 por cento). Os produtos de PCR indicaram a presença de Pneumocystis (21.56 por cento) em amostras obtidas de 8 das 16 espécies classificadas para ambos os estados (cinco morcegos não foram classificados). Este é o primeiro registro de detecção de Pneumocystis em morcegos no Brasil.

Pneumocystis diversity as a phylogeographic tool

Parasites are increasingly used to complement the evolutionary and ecological adaptation history of their hosts. Pneumocystis pathogenic fungi, which are transmitted from host-to-host via an airborne route, have been shown to constitute genuine host markers of evolution. These parasites can also provide valuable information about their host ecology. Here, we suggest that parasites can be used as phylogeographic markers to understand the geographical distribution of intra-specific host genetic variants. To test our hypothesis, we characterised Pneumocystis isolates from wild bats living in different areas. Bats comprise a wide variety of species; some of them are able to migrate. Thus, bat chorology and migration behaviour can be approached using Pneumocystis as phylogeographic markers. In the present work, we find that the genetic polymorphisms of bat-derived Pneumocystis are structured by host chorology. Therefore, Pneumocystis intra-specific genetic diversity may constitute a useful and relevant phylogeographic tool.

Comparison of methods for identification of Pneumocystis carinii in bronchoalveolar lavage fluid.

Pneumocystis carinii pneumonitis in one of the most common life-threatening opportunistic infections in patients with AIDS. The definitive diagnosis of this infection can be established only by demonstration of the organism in clinical specimens. This study was a comparison of methods that provide easy recognition of the organism which is readily available, simple and can be performed rapidly in laboratory-diagnosis. Bronchoalveolar lavage fluids obtained from 35 AIDS patients suspected of having Pneumocystis carinii pneumonitis were examined by three staining methods for the presence of Pneumocystis carinii. With Giemsa stains, P. carinii could be identified in 18 cases (51.4%). Three developmental stages: "cyst", "sporozoite" and "trophozoite" were seen. The contrast of organisms against host cells was not outstanding in these stains. Toluidine blue O stains provided easy recognition of the organisms, with marked contrast between the cysts and host cells. 21 cases (60%) were positive in these stains, but the intracystic structures and trophozoites could not be identified. It was suggested that the clinical specimen should be stained first with toluidine blue O which is more rapid and permits easy recognition of the cyst clusters. If the sporozoites and trophozoites had to be identified, Giemsa stains can be made. In addition, with the methenamine silver nitrate stains, 21 cases (60%) were positive. They revealed the morphology as seen with toluidine blue O but the cost of material may make it unavailable in many laboratories especially with the budgetary restraints of developing countries.

Pneumocystis carnii pneumonia in AIDS--an autopsy report.

Pneumocystis carinii pneumonia (PCP), the most common presenting manifestation in patients with AIDS from western countries, holds the distinction for being the first opportunistic infection that was associated with AIDS. There is marked paucity of clinically diagnosed and pathologically confirmed cases of PCP in India. This case represents the first complete autopsy report of pneumocystis carinii pneumonia inpatient with AIDS from our country. A high index of clinical suspicion and microscopic confirmation is needed to avert mortality due to PCP in patients with AIDS.

Neumonía por Pneumocistis carinii en pacientes con SIDA en El Hospital Santo Tomas / Pneumocistis carinii pneumonia in patients with AIDS at the Saint Thomas Hospital

The authors examined material from the respiratory tract obtained from 55 patients, with silver Methenamine and Giemsa stains to detect Pneumocystis carinii. Twenty five patients were positive. All had fever and fatigue, 80% had dyspnea, 72% had productive cough, and significant weight loss occurred in 48%. None of the patients had signs of pulmonary consolidation. Chest X-Ray showed diffuse infiltrates in 55.2%. Nodular lesions in both lung fields were present in 6.9%. A local infiltrate was seen in 13.6% and no infiltrates were seen in 10.3%. LDH was elevated in all (ranging from 885 4500 UI), 84% of the patients had a PO2 of 70 mmHg or less. Most of the patients, 80%, responded to therapy with Trimethoprim-Sulfa