Experimental paracoccidioidomycosis in the mouse. III. Histopathological and immunological findings after intravenous infection in the presence or absence of previous immunization

Cinqüenta camundongos suíços, brancos, com quatro semanas de idade, foram inoculados com 5x10(5) formas leveduriformes, viáveis de Paracoccidiodes brasiliensis (cepa 18). Dez destes animais tinham sido previamente imunizados com antígeno particulado de P. brasiliensis, durante quatro semanas, por injeção intradérmica. Os controles consistiram de 10 animais que foram somente imunizados e 10 inoculados com solução salina estéril. Os animais foram sacrificados após 2, 4, 7, 11 e 16 semanas. Estudamos: 1) resposta de hipersensibilidade retardada medida pelo teste do coxim plantar, 24 horas antes do sacrifício; 2) anticorpo- gênese específica avaliada pelo teste de imunodifusão dupla em gel de ágar; 3) histopatologia dos pulmões, fígado, baço, supra-renal e rins. Observamos: 1) os animais imunizados desenvolveram resposta imunecelular mais intensa que os infectados; 2) a infecção deprimiu a resposta imunecelular dos animais imunizados; 3) a histopatologia da infecção endovenosa revelou inflamação granulomatosa sistêmica e progressiva. Os animais infectados após imunização prévia apresentaram inflamação pulmonar menos extensa, com granulomas menores e com reduzido número de fungos. O presente modelo murino de paracoccidioidomicose mimetiza alguns achados da forma humana subaguda da micose (doença sistêmica com depressão da imunidade celular). O esquema de imunização extrapulmonar utilizado foi capaz de induzir certo grau de proteção do pulmão contra um desafio infeccioso pelo P. brasiliensis.

Road-killed wild animals: a preservation problem useful for eco-epidemiological studies of pathogens

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Uncommon mandibular osteomyelitis in a cat caused by Nocardia africana

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Time-place learning in individually reared angelfish, but not in pearl cichlid

Time-place learning based on food association was investigated in the cichlids angelfish (Pterophyllum scalare) and pearl cichlid (Geophagus brasiliensis) reared in isolation, therefore eliminating social influence on foraging. During a 30-day period, food was placed in one side of the aquarium (containing three compartments) in the morning and in the opposite side in the afternoon. Learning was inferred by the number of correct side choices of all fish in each day of test (15th and 30th). During the test day fish were not fed. The angelfish learned to switch sides at the correct day period in order to get food, suggesting this species has time-place learning ability when individually reared. on the other hand, the same was not observed for pearl cichlid. (c) 2006 Elsevier B.V. All rights reserved.

Animal welfare: from concepts to reality.

In this review, we provide a brief retrospective history of the science of animal welfare and recognize the sentience of non-human animals; however, we emphasize that crucial problems remain regarding how to define and measure animal welfare. In general, the use of physiological measures to assess welfare is discouraged. Furthermore, there is a theoretical background for measures of stress, but not for welfare states because life may not be at risk. Instead, a preference or choice-based approach, which is based on the animal decision, is recommended. To this end, welfare is discussed and then contrasted with disease, health, stress and distress. In addition, the importance of prospective capacities for the welfare of human and non-human animals is discussed.

Experimental paracoccidioidomycosis of the Syrian hamster: fungicidal activity and production of inflammatory cytokines by macrophages

Phagocytic cells play an important role in nonspecific resistance to fungal infection by mediating an inflammatory response and by a direct fungicidal action. In this study, the functional activity of peritoneal macrophages obtained from hamsters experimentally infected with strain Pb18 of Paracoccidioides brasiliensis was evaluated during 16 weeks of infection. The results showed that macrophages had a higher spreading ability associated with increased production of tumor necrosis factor alpha (TNF-alpha) and enhanced fungicidal activity during the early periods of infection. TNF-alpha levels remained elevated during all periods studied, while low levels of interleukin-1 beta (IL-1 beta) were produced during the infection. A necrotic area with dead fungi was observed at the inoculation site and the infection disseminated only to liver and lymph nodes in a few animals. These results suggest that during the early stages of infection with P. brasiliensis, macrophage activation by the high levels of TNF-alpha limited fungal dissemination. In contrast, in the later stages of infection, high levels of TNF-alpha were observed while the fungicidal activity of macrophages was lower and the animals presented loss of vitality resulting in their death. These observations suggest a complex role of TNF-alpha in experimental paracoccidioidomycosis of Syrian hamsters, involving not only resistance but also pathogenesis.

Inteins in pathogenic fungi: a phylogenetic tool and perspectives for therapeutic applications

Inteins or internal proteins are coding sequences that are transcribed and translated with flanking sequences (exteins). After translation, the inteins are excised by an autocatalytic process and the host protein assumes its normal conformation and develops its expected function. These parasitic genetic elements have been found in important, conserved proteins in all three domains of life. Most of the eukaryotic inteins are present in the fungi kingdom and the PRP8 intein is one of the most widespread inteins, occurring in important pathogens such as Cryptococcus neoformans (varieties grubii and neoformans), Cryptococcus gattii, Histoplasma capsulatum and Paracoccidioides brasiliensis. The knowledge of conserved and non-conserved domains in inteins have opened up new opportunities for the study of population variability in pathogenic fungi, including their phylogenetic relationships and recognition or diagnoses of species. Furthermore, inteins in pathogenic fungi should also be considered a promising therapeutic drug target, since once the autocatalytic splicing is inhibited, the host protein, which is typically vital, will not be able to perform its normal function and the fungal cell will not survive or reproduce.

Cytokine production in experimental paracoccidioidomycosis of murine selected lineages for acute inflammatory response (air)

Paracoccidioidomycosis is a systemic human mycosis caused by Paracoccidioides brasiliensis (P. brasiliensis), an imperfect dimorphic fungus whose conidia are its infective form. Mice genetically selected for maximum (AIRmax) and minimum (AIRmin) acute inflammatory response were used as experimental paracoccidioidomycosis models. The animals were intraperitoneally inoculated with P. brasiliensis (strain 18) and killed 6, 12 and 24 hours or 3, 7 and 14 days after infection. In these periods, fragments from their spleen, liver and lung were removed for evaluation of the infection level by fungal cells, assessment of macrophagic activity by peritoneal and splenic macrophages - through the determination of nitric oxide (NO) concentrations and production of pro- and anti-inflammatory cytokines of lung and spleen homogenate supernatants. In the present study, it was observed that AIRmax lineages presented greater control of the infectious process than the AIRmin ones. Regarding NO production, AIRmax animals produced more metabolites in late periods, what may help control the infectious process. Concerning cytokine production, it was observed that the production of INF-gamma, TNF-alpha, IL-1, IL-6, IL-8 and IL-12 were increased in AIRmax lineages in most analyzed organs and periods, thus contributing to the greater resistance exhibited by such lineages against infection, except for IL-4 and IL-10 that showed decreased production in AIRmax lineage, reproducing its suppressive biological effect. From these results, it was observed that the AIRmax lineage was more effective in controlling the infectious process, with an important involvement of the analyzed cytokines. These findings are probably related to the genetically selected factors involved in the acute inflammatory response.

PRP8 intein in Ajellomycetaceae family pathogens: Sequence analysis, splicing evaluation and homing endonuclease activity

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Phylogenetic analysis reveals a high level of speciation in the Paracoccidioides genus

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Spermatozoa ultrastructure in Sciaenidae and Polynemidae (Teleostei : Perciformes) with some consideration on Percoidei spermatozoa ultrastructure

Spermatozoa ultrastructure was studied in five marines (Paralonchurus brasiliensis, Larimus breviceps, Cynoscion striatus, Micropogonias furnieri, Menticirrhus americanus, Umbrina coroides, Stellifer rastrifer), and one freshwater (Plagioscion squamosissimus) species of Sciaenidae and one species of Polynemidae (Polydactylus virginicus). The investigation revealed that, in all species, spermatozoa display a round head, a nucleus containing highly condensed, filamentous chromatin clusters, no acrosome, a short midpiece with a short cytoplasmic channel, and a flagellum showing the classic axoneme structure (9 + 2) and short irregular lateral fins. In Sciaenidae, the spermatozoa are type II, the flagellar axis is parallel to the nucleus, the lateral nuclear fossa is double arched, the centriolar complex is outside the nuclear fossa, the proximal centriole is anterior and perpendicular to the distal centriole, and no more than ten spherical (marine species) or elongate (freshwater species) mitochondria are observed. Polynemidae spermatozoa are of the intermediate type with the flagellar axis eccentric to the hemi-arc-shaped nucleus, and exhibit no nuclear fossa, the centriolar complex close to the upper nuclear end, the proximal centriole lateral and oblique to the distal centriole, and one large ring-shaped mitocondrion. The data available show that no characteristic is exclusively found in the spermatozoa of members of the Sciaenidae family when compared to other Percoidei with type II spermatozoa. However, three characteristics were exclusively found in Polynemidae: (1) the hemi-arched nucleus; the positioning of the centrioles; and (2) the ring-shaped mitocondrion. The interrelationships between Sciaenidae and Polynemidae as well as between these two families and other Percoidei are herein discussed. (c) 2005 Elsevier Ltd. All rights reserved.

Genus Paracoccidioides: Species Recognition and Biogeographic Aspects

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Actinopterygii, Siluriformes, Loricariidae, Loricariichthys platymetopon Isbrucker and Nijssen, 1979: first record in Reservoir of Canoas II, Middle Paranapanema River, border of the states of São Paulo and Parana, Brazil.

First record of the cascudo, Loricariichthys platymetopon Isbrucker and Nijssen, 1979 in Canoas II Reservoir (Middle Paranapanema River, border of the states of São Paulo and Parana, Brazil). Its occurrence is due to the flooding of Sete Quedas waterfall, as a result of the construction of Itaipu Reservoir. Furthermore, there is absence or low abundance of natural predators, like Salminus brasiliensis, since this species has difficulties to adapt or even survive in reservoirs.

Helminths from Lizards (Reptilia: Squamata) at the Cerrado of Goias State, Brazil

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Larval development of Epialtus bituberculatus H. Milne Edwards, 1834 (Crustacea : Decapoda : Brachyura : Majidae) with comments on majid larvae from the southwestern Atlantic

The larval development of the spider crab Epialtus bituberculatus H. Milne Edwards which Lives on rocky shores with algae such as Sargassum and Hypneia, is described. Larvae were obtained from ovigerous females collected in Ubatuba, State of São Paulo, Brazil. Rearing was carried out at 24 +/- 1 degreesC, with an average salinity of 35 parts per thousand. Larval development consists of two zoeal stages and one megalopa. Zoeal development was completed in 9.5 days. Analysis indicated that zoeae of E. bituberculatus are very similar to those of E. brasiliensis Dana and Acanthonyx scutiformis (Dana). Differences noted between these species pertain to the setation of the carapace, maxillule and second maxilliped. The main morphological features useful for identification are presented together with a summary of features that characterize larvae of majid subfamilies in Brazil. A key for the identification of southwestern Atlantic majid zoeae to the family level is provided.