Anatomia de galhas de ambrosia em folhas de Baccharis concinna e Baccharis dracunculifolia (Asteraceae)

Galhas de ambrosia são induzidas por dípteros (Cecidomyiidae) e desprovidas de tecido nutritivo, pois a larva do indutor alimenta-se de hifas de fungos. As galhas de ambrosia de Baccharis concinna e B. dracunculifolia são constituídas por uma única câmara larval, contendo um indutor. São observadas hifas de fungos. Nas galhas de B. dracunculifolia, as hifas ficam confinadas à câmara larval e as células do parênquima paliçádico mostram-se alongadas. Nas galhas de B. concinna, as hifas estão presentes também entre as células do clorênquima situadas ao redor da câmara larval. As células do clorênquima próximas à câmara larval alongam-se ligeiramente. As fibras pericíclicas do sistema vascular, em ambas as galhas, perdem as paredes secundárias. Quando o indutor está em fase pupal, as hifas de fungos aumentam em quantidade e preenchem várias partes da câmara larval. Nas hifas da galha de B. concinna verifica-se a presença de glóbulos lipofílicos, que estão ausentes nas hifas das galhas de B. dracunculifolia. Picnídios são observados somente nas galhas senescentes de B. dracunculifolia. Este trabalho é a primeira contribuição ao conhecimento de galhas de ambrosia na flora brasileira.

Anatomical and developmental aspects of leaf galls induced by Schizomyia macrocapillata Maia (Diptera: Cecidomyiidae) on Bauhinia brevipes Vogel (Fabaceae)

Samples of healthy leaves and galls induced by Schizomyia macrocapillata Maia on Bauhinia brevipes Vogel were submitted to routine techniques to investigate gall anatomy and development. Pouch galls are induced on the abaxial surface of unfolded immature leaves, and become spheroid with long reddish hairs covering their external surface. Galls occur isolated or coalesce when in larger numbers. Gall development was divided into six phases: 1) initiation; 2) tissue re-arrangement; 3) tissue differentiation; 4) maturation; 5) growth phase; and 6) dehiscence. This last phase corresponds to gall senescence, which takes place just after the larva exits the chamber to pupate. An important developmental phase of tissue reorientation was recorded after the initiation phase. The presence of hyphae close to the covering layer characterizes this gall as an ambrosia gall and the feeding mode of the gall migde is discussed. Few hyphae were found during the first developmental phases and fungi may play an important role during gall morphogenesis. Neoformed trichomes may provide not only photoprotection but also protection against natural enemies and water loss. The neoformation of phloematic bundles suggests host manipulation and indicates the establishment of a deviating sink.

Immunization by subcutaneous implants of polyester-polyurethane sponges coupled with antigen

A new protocol is described for immunization of outbred Swiss mice. The procedure is based on subcutaneous implantation of antigen-coupled polyester-polyurethane sponges cut into disks of 10 mm in diameter vs 2 mm in thickness. Antigen coupling was performed by overnight incubation of the sponge with a solution of ovalbumin (Ova) (2 mg/ml) diluted in sodium carbonate buffer, pH 9.6. The amount of ovalbumin that was taken up by the sponge was between 71.4 to 82.5 µg. This was estimated by comparing the Ova absorbance at 280 nm in coating buffer solutions before and after incubation. To compare the efficiency of the proposed method, experimental groups immunized with the antigen in the presence of adjuvants (10 µg in Al(OH)3 or 100 µg in complete Freund's adjuvant (CFA)) were run in parallel. The data obtained after the 3rd week of immunization indicate that both cellular and humoral immune responses were achieved. These were assayed by antigen-induced footpad swelling and ELISA (specific antibodies), respectively. The levels of both immune responses elicited were similar to the responses observed in mice immunized with ovalbumin in the presence of Al(OH)3. The method might represent an advantage when immunizing with pathogenic antigens. Preliminary experiments have suggested that the antigen remains immobilized or bound to the sponge for a long period of time, since there is an increment on the cell population inside the sponges after boosting the animals. If so, the undesirable effects of immunization would be reduced.