Elizabeth Oliveira da Costa

Survey of bovine mycotic mastitis in dairy herds in the State of São Paulo, Brazil

The purpose of this study was to isolate fungi from the quarter milk of cow udders from several dairy herds and to identify the different genera and species involved in mastitis. A total of 2078 milk samples from normal, clinical and subclinical mastitis quarters from 22 dairy herds of 16 districts in the State of São Paulo, Brazil was utilized in this survey. Two hundred and fifty one (12.07%) fungi were isolated from the samples. Two hundred and eight of these (82.86%) were yeasts and 30 (11.95%) were moulds. The fungi were isolated in pure culture (24.77%) or in cultures mixed with bacteria (72.22%). The yeasts isolated were:Cryptococcus spp. (71 strains),Rhodotorula spp. (40),Candida spp. (68),Trichosporon cutaneum (21),Aureobasidium pullulans (7), andPichia ohmeri (1). Moulds classified in following genera were also isolated:Aspergillus (3),Penicillium (3),Alternaria (3),Phoma (3),Epicoccum (2), andGeotrichum (16).

Epidemiologic study of environmental sources in a Prototheca zopfii outbreak of bovine mastitis

Bovine mastitis represents the main form of occurrence of protothecosis in animals. The detection of mastitis caused by Prototheca sp. indicates a serious problem which can affect an entire herd. The purpose of this study is to explain some aspects of the epidemiology of mastitis due to Prototheca zopfii with the evaluation of the presence of these microorganisms in samples collected from potential sources in the dairy herd. This study was performed during a Prototheca zopfii outbreak of clinical bovine mastitis in the State of São Paulo, Brazil. The following samples were aseptically collected for microbiological examination: milk (n = 211); rectal swabs (from 15 calves and 2 lactating cows); swabs from teat cup rubbers during milking (n = 2); water (n = 6); soil (n = 6). Prototheca zopfii was isolated from 77 (36.49%) of the 211 milk samples; 11 calves and 2 cows showed Prototheca zopfii in faecal samples; both swabs collected from the teat cup rubbers showed viable forms of Prototheca zopfii; this microorganism was also isolated from 2 water samples, and 1 soil sample collected from the dry cow pasture. Prototheca zopfii seemed to be widespread throughout the dairy herd environment where this outbreak of bovine mastitis occurred.

Bovine mastitis due to algae of the genus Prototheca

Protothecosis was described in many animals, with bovine mastitis being the main form. The increasing number of isolations of Prototheca spp. from bovine mastitis cases indicates the need of a detailed evaluation of this problem. Besides this, these algae do not respond to treatment with the antimicrobians most frequently applied, leading to elimination of the affected animals, as the best method to control the disease. In two dairy farms in the State of São Paulo, Brazil, a total of 155 lactating cows and 52 dry cows were examined. Milk samples were aseptically collected from lactating cows with clinical or subclinical mastitis. From dry cows, secretion samples from all quarters were collected. All samples were then taken to microbiological exams. At dairy farm 1, Prototheca spp. was isolated from 14.95% milk samples from lactating cows, (all of them cases of subclinical mastitis), and from 8.06% samples from dry cows. At dairy farm 2, Prototheca spp. was isolated from 5.1% milk samples and there were ten cases of clinical mastitis due to this agent. Although clinical mastitis has been considered the main form of occurrence of this pathology until now, it is important to consider Prototheca spp. as subclinical mastitis pathogen. Prototheca zopfii was the main specie isolated. In this study a high rate of intramammarian infections, as clinical or subclinical mastitis during lactation as well as in dry period, was observed, high lighting the importance of suitable diagnosis, so that control and preventive measures can be implemented to avoid dissemination of the agent.

Evaluation of the susceptibility of Prototheca zopfii to milk pasteurization

Protothecosis has been reported in humans (gastroenteritis, bursitis, etc.) and in many other animal species. Bovine mastitis represents the main form of occurrence of protothecosis in cattle. Milk as well as dairy products, when contaminated with Prototheca spp., represent a potential means of transmission of this zoonosis. The purpose of this study was to evaluate the susceptibility of forty Prototheca zopfii strains isolated from milk from intramammary infections in dairy cows and also from bulk milk tanks of dairy farms, to the different ratios of temperature/time employed in the thermal treatment of milk: 72–75 °C/1 5 seconds, 72–75 °C/20 seconds and 62–65 °C/30 minutes. The samples were subjected to these different temperature/time ratios. The evaluation of the thermal susceptibility of the P. zopfii strains showed that 34 strains were resistant in at least one of the tests. The results point out the need to consider the importance of mastitis caused by Prototheca spp. asrepresenting a public health risk.

Clinical disorders observed in anteaters (Myrmecophagidae, Edentata) in captivity.

The major health problems found in 103 captive lesser anteaters (Tamandua tetradactyla) and giant anteaters (Myrmecophaga tridactyla), family Myrmecophagidae, are presented and correlated with management. The most common of 200 recorded clinical disorders involved the digestive system (26%), nutritional deficiency (20%), injury (15.5%), respiratory system (10%), skin (7%) and circulatory system (4.5%), but 13% of the cases were inconclusive. Parasites were identified in 48.5% of faecal samples, mainly the eggs of nematodes (40%), of which the commonest wereTrichuris spp (28%) andStrongyloides spp (11%); protozoa (16%), of which the commonest wereEimeria spp (10%),Entamoeba spp (5%) andGiardia spp (1%); and cestodes (8%) and acanthocephalids (1%). Bacteria cultured from the various materials includedSalmonella enteritidis, S. cholerasuis, Escherichia coli, Enterobacter aerogenes, Streptococcus spp andStaphylococcus spp. The ectoparasites found wereAmblyomma spp andOtodectis spp (Arthropoda, Acaridae).

Delayed hypersensitivity test with paracoccidioidin in captive Latin American wild mammals

The aim of this investigation was to study epidemiological aspects of paracoccidioidomycosis, the main endemic systemic mycosis in Brazil. This study was carried out using the paracoccidioidin delayed hypersensitivity test in 96 Latin American wild mammals, including 49 arboreal animals (primates): 33 Cebus apella (weeping-capuchin), 16 Callithrix jacchus (marmoset); and 47 terrestrial animals (carnivora): 37 Nasua nasua (coatimundi), and 10 Felidae [Panthera onca (jaguar), Felis paradalis (ocelot), Felis wiedii (margay), Felis tigrina (wild cat) and Felis geoffroyi (wild cat)], taking their behaviour and habitat into consideration. When the levels of paracoccidiodin positive reactions were examined, terrestrial animals showed significantly higher rates (82.98%) while arboreal animals showed lower reactivity (22.45%) (P < 0.01). The data are relevant because there are quite a few papers regarding domestic and wild animals and this study may help the understanding of some aspects of the parasite ecology. These results point to the soil as the most probable reservoir of Paracoccidioides brasiliensis, and this is possibly the ecological niche of the saprophytic phase in nature.

Epidemiological study of sporotrichosis and histoplasmosis in captive Latin American wild mammals, São Paulo, Brazil

Sporotrichosis and histoplasmosis are deep mycosis with a high incidence in human beings in Brazil. In domestic animals histoplasmosis has been described only in dogs, but the occurence of sporotrichosis among domestic animals in Brazil has been described in dogs, cats, mules and asses. There is also a case of this disease reported in a chimpanzee (Pan troglodites). The purpose of this research was to perform an epidomiological study of these mycoses using delayed hypersensitivity tests (histoplasmin and sporotrichin) in Latin American wild mammals. This research was assayed using 96 healthy animals at Parque Zoológico de São Paulo, Brazil: Primates: 33Cebus apella — weeping-capuchin and 16Callithrix jacchus — marmoset; Procyonidae: 37Nasua nasua — coatimundi and 10 Felidae (Panthera onca — jaguar;Felis pardalis — ocelotFelis wiedii — margay;Felis tigrina — wild cat). For intradermic tests, the following antigens were used:Sporothrix schenkii cell suspension (sporotrichin, histoplasmin-filtrate),Histoplasma capsulatum cell suspension (histoplasmin), andHistoplasma capsulatum (polysaccharide). The positivity to histoplasmin was 44.79% (Cebidae 15.15%; Callithricidae 6.25%; Procyonidae 86.49% and Felidae 50.00%, respectively). With respect to sporotrichin, 30.21% (Cebidae 6.06%, Callithricidae 0.0%; Procyonidae 64.86% and Felidae 30.00% respectively). The pattern of infection is similar to that shown by human beings and this may suggest that these animals could be involved in the epidemiologic chain of sporotrichosis and histoplasmosis, the second most prevalent human deep mycoses in Brazil. It is important to point out the absence of similar studies in Latin American wild animals.

Susceptibility and features of the ultrastructure of Prototheca zopfii following exposure to copper sulphate, silver nitrate and chlorexidine

One of the most important forms of the occurrence of protothecosis is bovine mastitis. Studies on the “in vivo” and “in vitro” susceptibility to antimicrobials have shown that the microorganism is resistant to most of them. Looking for alternative treatments this study aimed to study the susceptibility to copper sulphate (which has an important algicide effect) and silver nitrate (used in dairy cattle breeding for the cauterization of mammary glands) and also to chlorexidine (an important post-dipping anti-septic used in dairy practice), and the effect of these antimicrobials in the ultrastructure of Prototheca zopfii before and after the exposure to these drugs. The “in vitro” susceptibility tests to chlorexidine, silver nitrate and copper sulphate of the strains of Prototheca zopfii for the determination of their minimal microbicidal concentrations (MMC), were performed using the tube dilution method in Sabouraud dextrose broth and evaluation of colony growth after plating in Sabouraud dextrose agar. The MMCs of chlorexidine, copper sulphate and silver nitrate of the 50 strains tested were 0.01%, 0.1% and 0.3%, respectively. The tubes containing the material used in the antimicrobial susceptibility tests were prepared for the examination in an electron microscope. The untreated controls of P. zopfii showed a similar ultrastructural appearance with the typical characteristics of the microorganism. Cells exposed to silver nitrate showed changes suggesting thickness of the cell wall. Cells exposed to chlorexidine showed changes suggesting degradation of intra-cellular organelles present in the cytoplasm. P. zopfii treated with copper sulphate showed changes suggesting fibrilation of inner layer of cell wall.

The prevalence of positive intradermal reactions to paracoccidioidin in domestic and wild animals in São Paulo, Brazil

Costa, E.O., Diniz, L.S.M. and Fava Netto, C., 1995. The prevalence of positive intradermal reactions to paracoccidioidin in domestic and wild animals in Silo Paulo, Brazil. Veterinary Research Communications, 19 (2), 127-130

Contribution to the epidemiology of paracoccidioidomycosis and histoplasmosis in the state of São Paulo, Brazil: Paracoccidioidin and histoplasmin intradermic tests in domestic animals

Bovines, ovines and equines from various counties in the state of Sao Paulo, Brazil, show a high incidence of paracoccidioidomycosis infection according to the results obtained with intradermic test standardized in experimentally infected bovines: 44·88%, 42·86% and 77·06% respectively for bovines, ovines and equines.The antigen chosen for this study was a suspension of P. brasiliensis yeast cells after extraction of lipidic substances by acetone and ether and extraction of the polysaccharide antigen by autoclavation. The volume used for the intradermic tests was 0·2 ml for the bovines and equines and 0·1 ml for the ovines. These quantities permitted formation of well-defined papules, indicating that the antigen had been correctly injected intradermically.Our results indicate that two readings should be taken, 24 and 48 hours after injection of the antigen. If only one reading is possible, the 24 hours reading is preferable.Positive reaction to the paracoccidioidin test was of the same magnitude in bovine...