Edna Maria Cavallini Sanches

Diversity of yeasts from bovine mastitis in Southern Brazil.

Mastitis is one of the most serious problems in the dairy cattle farms. The great majority of the cases are caused by bacteria, but lately there have been an increasing number of reports about cases of mycotic etiology. The objective of this work was to characterize the yeasts and yeast-like fungi associated with milk of cows with mastitis. Milk samples (n = 248) from a dairy belt situated around the region of Passo Fundo, hinterland of the state of Rio Grande do Sul, Southern Brazil, were analyzed. Aliquots of 0.1 ml of milk were inoculated on yeast extract-malta agar with chloramphenicol. After a period of incubation of 3-5 days at 22-25 degrees C, the counting of the morphologically distinct colonies was performed, as well as the isolation and identification through phenotypical and physiological criteria. It was possible to isolate 68 yeast species from 43 (17.3%) of the samples. The most frequent genera were Candida (37.9%), Pichia (19.1%), Cryptococcus (10.3%) and Rhodotorula (10.3%).

Co-infection of Pneumocystis carinii f. sp. suis and porcine circovirus-2 (PCV2) in pig lungs obtained from slaughterhouses in southern and midwestern regions of Brazil.

EDNA M. CAVALLINI SANCHES, MAURO R. BORBA, ANDRÉIA SPANAMBERG, CAROLINE PESCADOR, LUÍS G. CORBELLINI, ANA P. RAVAZZOLO, DAVID DRIEMEIER and LAERTE FERREIRO Sections of Veterinary Mycology, Departamento de Patologia Clı́nica Veterinária, Faculdade de Veterinária, Universidade Federal do Rio Grande do Sul (UFRGS), Av. Bento Gonçalves 9090, 90540-000, Porto Alegre, RS, Brazil, and Sections of Pathology, Departamento de Patologia Clı́nica Veterinária, Faculdade de Veterinária, Universidade Federal do Rio Grande do Sul (UFRGS), Av. Bento Gonçalves 9090, 90540-000, Porto Alegre, RS, Brazil, and Sections of Immunology, Departamento de Patologia Clı́nica Veterinária, Faculdade de Veterinária, Universidade Federal do Rio Grande do Sul (UFRGS), Av. Bento Gonçalves 9090, 90540-000, Porto Alegre, RS, Brazil

Mastite micótica em ruminantes causada por leveduras

The veterinary literature registers sporadic cases of infections caused by environmental microorganisms in which the main agents involved are yeasts, yeasts-like and filamentous fungi. In relation to dairy animals, the yeasts are more frequently incriminated as the aetiological agents of mycotic mastitis. Most cases occur under the form of localized outbreaks and / or after treatment with antimicrobial agents. The main genera involved in mastitis are Candida and Cryptococcus, besides others such as Geotrichum, Pichia and Trichosporon. The purpose of this review is to point out the main aspects of mastitis caused by yeasts and yeast-like fungi, such as etiology, predisposing factors, pathogenicity, diagnosis, treatment and prophylaxis, through a chronological approach of the first reports, with main focus on results described recently in national and international veterinary literature. The mycotic mastitis control should be mainly focused on preventive methods, especially based on adequate management of essential factors like proper milking procedures and a high level of hygiene of the environment and equipments, with the aim of reducing the number of affected animals, assuring the quality and innocuity of dairy food safety.

Detection of Pneumocystis in lungs of bats from Brazil by PCR amplification

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.

Immunohistochemical and ultra-structural detection of Pneumocystis in wild boars (Sus scrofa) co-infected with porcine circovirus type 2 (PCV2) in Southern Brazil

Pneumocystis spp. are fungi that are able to infect a variety of host species and, occasionally, lead to severe pneumonia. Porcine circovirus type 2 (PCV2) is an important viral pathogen which affects both swine and wild boar herds worldwide. Co-infection between PCV2 and other pathogens has been reported, and the secondary immunodeficiency caused by the virus may predispose to these co-infections. In the present study, postmortem tissue samples obtained from wild boar herds in Southern Brazil were analyzed by histopathology, ultra-structural observation, and immunohistochemistry. Forty-seven out of seventy-eight (60%) wild boars showed clinical signs, gross, and histopathological lesions characteristic of infection by PCV2. Pneumocystis was detected by immunohistochemistry in 39 (50%) lungs and viral antigens of PCV2 were found in 29 (37.2%) samples. Concomitant presence of Pneumocystis and PCV2 were observed in 16 (20.5%) of the wild boars. Cystic and trophic forms of Pneumocystis were similar to previously described ultra-structural observations in other mammals.

Phylogenetic analysis of Pneumocystis from pig lungs obtained from slaughterhouses in southern and midwestern regions of Brazil

The Pneumocystis genus is comprised of pathogens dwelling in the lungs of terrestrial, aerial, and aquatic mammals. Occasionally they induce severe pneumonitis, particularly in hosts with severe impairment of the immune system and progressively may fill pulmonary alveolar cavities causing respiratory failure. Molecular genetic studies revealed that Pneumocystis gene sequences present a marked divergence with the host species concerned. In the present study, the genetic diversity of Pneumocystis obtained from lungs of swines was examined by analyzing mitochondrial large subunit (mtLSU) and small subunit (mtSSU) rRNA sequences. The samples were obtained from two slaughterhouses located in two Brazilian states. Phylogenetic analysis demonstrated that genetic groupings within Pneumocystis organisms were in accordance with those of the corresponding hosts and that two clusters were formed. In conclusion, these data show that there are genetically distinct porcine Pneumocystis genotypes with at least two separate clusters in Brazil.

Efeito do congelamento sobre a viabilidade de células leveduriformes

O controle de produtos de origem animal esta diretamente associado a sanidade animal e a saude publica. Na suspeita de mastite, tanto bacteriana quanto micotica, o congelamento de amostras de leite permite aumentar o periodo de tempo para analise microbiologica, permitindo um correto diagnostico laboratorial. As leveduras sao os principais agentes ambientais envolvidos na mastite micotica. O objetivo do presente trabalho foi verificar se o congelamento afeta a recuperacao 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). Apos a inoculacao experimental, as amostras foram congeladas durante 10 dias, 3, 6 e 8 semanas. O congelamento do leite por periodos superiores a 10 dias causou reducao no numero de celulas viaveis. Sugerimos que sejam utilizados periodos mais curtos de congelamento para a analise micologica do leite.