Andréia Spanamberg

In Vitro Activities of Voriconazole, Itraconazole, and Terbinafine Alone or in Combination against Pythium insidiosum Isolates from Brazil

ABSTRACT We evaluated the in vitro activities of voriconazole, itraconazole, and terbinafine against 30 clinical isolates of Pythium insidiosum using a checkerboard macrodilution method. The combined activity of terbinafine plus itraconazole or plus voriconazole was synergic against 17% of the strains. Antagonism was not observed.

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

Detection of Pneumocystis spp. in lung samples from pigs in Brazil

The genus Pneumocystis is composed of opportunistic fungi currently considered as specific pulmonary pathogens in humans and other mammals. In pigs, Pneumocystis pneumonia (PcP) could create significant economical losses due to its detrimental effects on growth, food conversion, and carcass/viscera condemnation. This study revealed that Pneumocystis organisms could be detected by Grocotts staining or immunohistochemistry in 36.9% of 564 slaughtered pigs from two geographic regions of Brazil. The prevalence of positive cases was 39.9% and 33.9% in pigs slaughtered in Rio Grande do Sul (RS) and Mato Grosso (MT) states, respectively. Among the positive cases in RS, Pneumocystis organisms were observed in 41.9% of 33 histologically normal lungs, and in 58.0% of lungs presenting with histological lesions. In contrast, the prevalence in MT in normal and abnormal lungs was 36.3% and 63.5%, respectively. Major histopathological findings in lungs of infected animals were bronchointerstitial pneumonia (47.6%), suggestive of enzootic pneumonia, and interstitial pneumonia (37.9%), compatible with PcP. The results of this survey strengthened the interest of detecting fungal pathogens, in addition to other infectious agents, and evaluating their financial impact on Brazilian pig industry. Preventive and/or therapeutic strategies should be developed in order to minimize the incidence of respiratory fungal infections in pigs and associated economic losses.

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.

In vitro paradoxical growth of Pythium insidiosum in the presence of caspofungin

Pythium insidiosum is a zoosporic organism which causes pythiosis in humans and animals. This study aimed to report the paradoxical growth of Brazilian P. insidiosum strains when submitted to in vitro susceptibility tests with caspofungin. The growth at concentrations above the minimal inhibitory concentration (MIC) ranged from 16 to 128 μg/ml and it was observed in 50% of the isolates tested. This paradoxical growth in the presence of caspofungin has been observed with Candida and Aspergillus strains, however, the phenomenon involving oomycetes was described here for the first time.

Aspergillus fumigatus from normal and condemned carcasses with airsacculitis in commercial poultry

Nos abatedouros, a inspecao das carcacas e fundamental para a deteccao e monitoramento da prevalencia de certas doencas. Os objetivos do trabalho foram avaliar a ocorrencia de aspergilose causada por Aspergillus fumigatus em aves comerciais atraves do diagnostico micologico e histopatologico e verificar a possibilidade de associacao causal entre os criterios de diagnostico de aspergilose e condenacao por aerossaculite em frangos de corte atraves de um estudo de caso-controle. O estudo foi realizado com 380 amostras. Foram coletados pulmoes de frangos condenados (95) e nao condenados (285) por aerossaculite, diretamente na linha de abate de um frigorifico. Quarenta e seis (12%) amostras de pulmao foram positivas na cultura micologica. Do total de amostras, 177 (46,6%) apresentaram alteracoes histopatologicas, sendo os mais frequentes pneumonia fibrinoheterofilica necrotica, pneumonia heterofilica e hiperplasia linfoide. Do total de 380 pulmoes analisados, 65,2% (30) apresentaram alteracoes histopatologicas e isolamento fungico. A relacao entre a presenca de lesoes histopatologicas e isolamento de A. fumigatus testada por McNemar indicou que houve associacao significativa entre a presenca de alteracoes histopatologicas e o isolamento de A. fumigatus. O cultivo micologico e o exame histopatologico aumentam as chances de se detectar alteracoes pulmonares em aves condenadas pelo Sistema de Inspecao Final do que nas aves normais, sugerindo que tais criterios de diagnostico sao eficazes para aprimorar a avaliacao e condenacao de aves por aerossaculite.

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.

Enzymatic variability among Brazilian Pythium insidiosum isolates.

BACKGROUND Pythium insidiosum is an oomycete classified in the kingdom Stramenopila. P. insidiosum hyphae are not able to initiate infection without the secretion of hydrolytic enzymes, which are considered an important factor in microbial virulence. AIMS To evaluate the extracellular enzymatic activity of 14 Brazilian P. insidiosum isolates and a standard strain (ATCC 58637) by the API-ZYM System screening method. METHODS Zoospores were grown in RPMI 1640 broth, and 65 μL of the liquid phase were inoculated in each cupule of the API-ZYM strips. RESULTS Differences in the enzymatic activities were observed among the isolates, although phosphohydrolases and ester hydrolases were conspicuous among all isolates. β-glucosidase was also present in most of the isolates. Enzymatic activities of α-glucosidase and chymotrypsin were not observed, differing from a previous study involving Australian isolates and intracellular enzymes. CONCLUSIONS The discrepancy in the enzymatic profile observed among Brazilian P. insidiosum isolates reflects the phenotypic variations found in susceptibility tests.