Influence of experimental illumination and seasonal variation on crossbreending mating in the snail Biomphalaria Glabrata

The crossbreeding activities of the Schistosoma mansoni vector snail Biomphalaria glabrata were counted in a laboratory aquarium throughout the year under two regimes of 12h light: 12h dark from 7 A., M. to 10 P. M. Mating increased significantly in Authmn and Winter and just missed a significant inverse correlation with temperature and a direct one with locomotion. Other similar experiments were carried out to compare mating under various ilumination conditions in complete daily cycle measurements. Mating counts decreased under the regimes which submited snail to a total exposure of 12h light and 12 dark during a daily cycle in the following sequence: 12h light: 12h dark alternating hourly with light gradient, 12h light: 12h dark, 1h light: 1h dark and 12h dark: 12h light. Under two constant illuminations, the mating scored less than under the previous conditions, except under 12h light. Under darkeness the mating count was lower than light conditions. There was no way to differentiate the night and day rhythms of mating on different days in each regime, except for mating under 12h light: 12 dark alternating with light gradient, constant dark and 12h dark: 12h light conditions. Mating increased in certain light and temperature conditions, in wich the intensities, should have an optimum value.

Changes in the behavioral and immunological parameters of the mollusk Biomphalaria tenagophila induced by disruption of the circadian cycle as a consequence of continuous illumination

In the present investigation we studied some behavioral and immunological parameters of adult gastropod mollusk, Biomphalaria tenagophila, which have been reproducing for several generations under laboratory conditions. One group of gastropods was kept on a 14-h light/10-h dark cycle, corresponding to a regular circadian cycle, and another group was exposed to continuous light for 48 h. Animals were studied along (behavioral groups) or immediately after (immunological groups) 48 h of regular circadian cycle or continuous light conditions. Stopping/floating, dragging and sliding were the behavioral aspects considered (N = 20 for regular cycle; N = 20 for continuous illumination) and number of hemocytes/µl hemolymph was the immunological parameter studied (N = 15 for regular cycle, N = 14 for continuous illumination). Animals under continuous illumination were more active (sliding = 33 episodes, dragging = 48 episodes) and displayed a lower number of hemocytes (78.0 ± 24.27/µl) when compared with mollusks kept on a regular circadian cycle (sliding = 18 episodes, dragging = 27 episodes; hemocytes = 157.6 ± 53.27/µl). The data are discussed in terms of neural circuits and neuroimmunological relations with the possible stressful effect of continuous illumination.

Th1 polarized response induced by intramuscular DNA-HSP65 immunization is preserved in experimental atherosclerosis

We previously reported that a DNA vaccine constructed with the heat shock protein (HSP65) gene from Mycobacterium leprae (DNA-HSP65) was protective and also therapeutic in experimental tuberculosis. By the intramuscular route, this vaccine elicited a predominant Th1 response that was consistent with its protective efficacy against tuberculosis. It has been suggested that the immune response to Hsp60/65 may be the link between exposure to microorganisms and increased cardiovascular risk. Additionally, the high cholesterol levels found in atherosclerosis could modulate host immunity. In this context, we evaluated if an atherogenic diet could modulate the immune response induced by the DNA-HSP65 vaccine. C57BL/6 mice (4-6 animals per group) were initially submitted to a protocol of atherosclerosis induction and then immunized by the intramuscular or intradermal route with 4 doses of 100 µg DNA-HSP65. On day 150 (15 days after the last immunization), the animals were sacrificed and antibodies and cytokines were determined. Vaccination by the intramuscular route induced high levels of anti-Hsp65 IgG2a antibodies, but not anti-Hsp65 IgG1 antibodies and a significant production of IL-6, IFN-g and IL-10, but not IL-5, indicating a Th1 profile. Immunization by the intradermal route triggered a mixed pattern (Th1/Th2) characterized by synthesis of anti-Hsp65 IgG2a and IgG1 antibodies and production of high levels of IL-5, IL-6, IL-10, and IFN-g. These results indicate that experimentally induced atherosclerosis did not affect the ability of DNA-HSP65 to induce a predominant Th1 response that is potentially protective against tuberculosis.

SPAD-502 readings in response to photon fluence in leaves with different chlorophyll content

The chlorophyll meter (SPAD-502) is widely used to estimate chlorophyll content, but non-uniform chloroplast distribution can affect its accuracy. This study aimed to assess the effect of photon fluence (F, irradiance x time of illumination) in leaves with different chlorophyll content and determine the effect of chlorophyll a/b on SPAD values of four tropical tree species (Croton draconoides Müll. Arg., Hevea guianensis Aubl., Hymenaea courbaril L. and Matisia cordata H.B.K.). There were also determined calibration equations for the chlorophyll meter and assessed the effect of F on SPAD values between 07:00 h and 17:00 h. Calibration equations were obtained after determining leaf chlorophyll content in the laboratory. Increases in F with time caused a reduction in SPAD values in species with a high chlorophyll content, with reductions of 20% in M. cordata and 10% in H. guianensis. Leaves of C. draconoides and H. courbaril had lower chlorophyll content and showed no changes in SPAD values with increase in F. The chlorophyll a/b ratio increased with SPAD values and the SPAD/chlorophyll relationship was best described by an exponential equation. It seems that F may affect SPAD values in leaves with high chlorophyll content, probably due to non-uniform chloroplast distribution at high irradiance. This indicates that SPAD values tend to be more accurate if recorded early in morning when irradiance is low.

APPLICATION OF 6-NITROCOUMARIN AS A SUBSTRATE FOR THE FLUORESCENT DETECTION OF NITROREDUCTASE ACTIVITY IN Sporothrix schenckii

Introduction Sporothrix schenckii is a thermal dimorphic pathogenic fungus causing a subcutaneous mycosis, sporotrichosis. Nitrocoumarin represents a fluorogenic substrate class where the microbial nitroreductase activity produces several derivatives, already used in several other enzyme assays. The objective of this study was the analysis of 6-nitrocoumarin (6-NC) as a substrate to study the nitroreductase activity in Sporothrix schenckii. Methods Thirty-five samples of S. schenckii were cultivated for seven, 14 and 21 days at 35 °C in a microculture containing 6-nitrocoumarin or 6-aminocoumarin (6-AC) dissolved in dimethyl sulfoxide or dimethyl sulfoxide as a negative control, for posterior examination under an epifluorescence microscope. The organic layer of the seven, 14 and 21-day cultures was analyzed by means of direct illumination with 365 nm UV light and by means of elution on G silica gel plate with hexane:ethyl acetate 1:4 unveiled with UV light. Results All of the strains showed the presence of 6-AC (yellow fluorescence) and 6-hydroxylaminocoumarin (blue fluorescence) in thin layer chromatography, which explains the green fluorescence observed in the fungus structure. Conclusion The nitroreductase activity is widely distributed in the S. schenckii complex and 6-NC is a fluorogenic substrate of easy access and applicability for the nitroreductase activity detection.

Contribution to the histology of Biomphalaria glabrata

A combination of histological techniques applied to the study of Biomphalaria glabrata yielded some interesting new data about the histology of this snail, a major intermediate host of Schistosoma mansoni in Brazil. Three kinds of pigments were identified: a dark pigment which bleached following oxidation with potassium permanganate; a lipofuchsin-like, diastase-resistant PAS-positive pigment and an iron-containing pigment, probably related to hemosiderin. Calcium was detected in small deposits within the connective tissue and forming a dense core inside the chitinous radular teeth. The presence of fibrils, staining with sirius-red and birefringence under polarized light strongly suggest primitive collagen tissue. The radular apparatus appeared as a storing site for glycogen, while abundant Alcian-blue positive material (proteoglycans) was extremely concentrated in the radular sac.

Comparison of the fluorescent polarization (TDx) and the enzymatic competitive (EMIT 2000) immune assays for the measurement of cyclosporin A blood concentration

Evaluation of Cyclosporin A (CyA) blood concentration is imperative in solid organ transplantation in order to achieve maximal immunosuppression with the least side effects. We compared the results of whole blood concentrations of CyA in 50 blood samples simultaneously evaluated by the fluorescent polarization immune assay (TDx) and the enzymatic competitive immune assay (EMIT 2000). There was a strong correlation between both kits for any range of CyA blood concentration (R=0.99, p<0.001). The within-run and between-days coefficient of variation were less than 4% for both assays. The cost for each CyA measurement was 50% lower for the EMIT assay when compared to the TDx assay. We concluded that the EMIT is as accurate as the TDx in measuring CyA blood concentration and has the advantage of a lower cost, as well as the possibility of widespread access to the EMIT methodology in contrast to the TDx equipment, allowing the laboratory to perform several routines within a working day.

Titanium pigment in tissues of drug addicts: report of 5 necropsied cases

The aim of this report is to describe the anatomic-pathologic findings from necropsies of 5 drug addicts with titanium pigment in several organs after chronic intravenous injection of crushed propoxyphene hydrochloride tablets. Samples from liver, spleen, lungs, lymph nodes, and bone marrow were obtained, and after being grossly studied, they were submitted to evaluation using common light and polarized microscopy. In all 5 cases, a pigment with characteristics of titanium dioxide was found within tissue samples of the organs studied. Our findings suggest that research concerning titanium pigment within body tissues should be enhanced, considering the potential contribution of this morphologic data to forensic pathology.

Multi-temporal analysis of radiometric changes in satellite images of forest edges to infer selective-logging areas in the Amazon forest

Radiometric changes observed in multi-temporal optical satellite images have an important role in efforts to characterize selective-logging areas. The aim of this study was to analyze the multi-temporal behavior of spectral-mixture responses in satellite images in simulated selective-logging areas in the Amazon forest, considering red/near-infrared spectral relationships. Forest edges were used to infer the selective-logging infrastructure using differently oriented edges in the transition between forest and deforested areas in satellite images. TM/Landsat-5 images acquired at three dates with different solar-illumination geometries were used in this analysis. The method assumed that the radiometric responses between forest with selective-logging effects and forest edges in contact with recent clear-cuts are related. The spatial frequency attributes of red/near infrared bands for edge areas were analyzed. Analysis of dispersion diagrams showed two groups of pixels that represent selective-logging areas. The attributes for size and radiometric distance representing these two groups were related to solar-elevation angle. The results suggest that detection of timber exploitation areas is limited because of the complexity of the selective-logging radiometric response. Thus, the accuracy of detecting selective logging can be influenced by the solar-elevation angle at the time of image acquisition. We conclude that images with lower solar-elevation angles are less reliable for delineation of selecting logging.

The relation of the concentration of macronutrients in the substrate and in the foliage to cell wall thickness and cellulose concentration in the xylem of slash pine (Pinus elliotti)

Sand culture experiments, using a sub-irrigation technique, were installed in order to find out the effects of the macronutrients N, P, K, Ca, Mg and S on growth, aspect, mineral composition, length of fibers, thickness of cell wall and cellulose concentration in slash pine. The aim was to obtain, under controlled conditions, basic information which could eventually lead to practical means designed to increase the rate of growth and to make of slash pine a richer source of cellulose. Nitrogen, Phosphorus, Potassium Experiment A 3 x 3 x 3 factorial design with two replicates was used. Nitrogen was supplied initially at the levels of 25, 50 and 100 ppm; phosphorus was given at the rates of 5, 10 and 20 ppm; potassium was supplied at the rates of 25, 50 and 100 ppm; six months after the experiment was started the first level for each element was dropped to zero. Others macro and all micronutrients were supplied at uniform rates. Fifteen hours of illumination per day were provided. The experimental technique for growing the slash pine seedlings proved quite satisfactory. Symptoms of deficiency of nitrogen, phosphorus and potassium were observed, described and recorded in photographs and water colors. These informations will help to identify abnormalities which may appear under field conditions. Chemical analysis of the several plant parts, on the other hand, give a valuable means to assess the nutritional status of slash pine, thus confirming when needed, the visual diagnosis. The correctness of manurial pratices, on the other hand, can be judged with the help of the analytical data tabulated. Under the experimental conditions nitrogen caused the highest increases on growth, as measured by increments in height and dry weights, whereas the effects of phosphorus and potassium were less marked. Cellulose concentration was not significantly affected by the treatments used. Higher levels of N seemed to decrease both length of fiber elements and the thickness of cell wall. The effects of P and K were not well defined. Calcium, Magnesium, Sulfur Experiment A 3 x 3 x 3 factorial design with two replicates was used. Calcium was supplied initially at the levels of 12.5, 25 and 50 ppm; magnesium and sulfur were given at the rates of 6, 12.5 and 25 ppm. Other macro and micronutrients were supplied at uniform rates, common to all treatments. Three months after starting the experiment the first level for each element was dropped to zero. Symptoms of deficiency of calcium, magnesium and sulfur were observed, described and recorded as in the case of the previous experiment. Chemical analysis were made, both for mineral content and cellulose concentration. Length of fibers and thickness of cell wall were measured. Both calcium and magnesium increase height, sulfur failing to give significant response. Dry weight was beneficially affected by calcium and sulfur. The levels of calcium, magnesium and sulfur in the needles associated with deficiency and maximum growth are comparable with those found in the literature. Cellulose concentration increased when the level of sulfur in the substrate was raised. The thickness of cell wall was negatively affected by the treatments; no effect was observed with regards to length of fibers.

Biologia do saguirú (Characidae, Curimatinae)

In Brazil all the fishes belonging to the sub-family Curimatinae are called « saguirú ». The present work gives a biological study of the Curimatus elegans Steind., a small fish without any economical importance, which is to be found along the whole brazilian coast, down till Paraguay. The specimens utilized for the present study come from Fortaleza (Ceará, north-eastern Brazil). The C. elegans is « ilyophagus », that means, it feeds itself exclusively with those organic materials to be found in mud, specially with microscopical algae. The intestines are very extent, some of them measuring about 9 to 11 times body's length. Studies have been made about growth and age of the C. elegans; the biggest sizes found were of 153 mm. for females and 88 mm. for males. The C. elegans shows developed sexual glands during a long period (April to September). The movements of the spermatozoa, in contact with water is of 40 to 50 seconds of intense movements, ceasing after 70 to 100 seconds. In contact with 0.5% NaCl-solution spermatozoa show a big increase in movements-time, that can last till about 25 minutes. The eggs' diameter measures 0.70 to 0.73 mm., mature and hydrated it attains 0.93 to 1,00 mm. There is a certain correlation between the size of the body and the quantity of eggs. Big specimens can produce a total of 200.000 eggs. The average quantity contained in 1 gr. and 1 cc. is 6018 and 6229 eggs, respectively. Maturity and spawning in laboratory has been obtained due to injections of suspension of fish-hypophysis. Three or four hours after the injection, fishes show more movement and evident signs of excitation, proceeding spawning after 5 to 6 hours. Males, persecuting females, describe successive circles (merry-go-round) - carroussel), swimming side by side with females up to water's surface, where sexual products are start beating dry, for there is no blood yet. Circulation-scheme is to be found on fig. 4 and 5. The swim-bladder and the stomach are but delineated; the intestine is formed by a cylindric tube, all closed. At the place, where later on there will open the mouth, we find a group of ciliary hairs that produce a liquid current, very evident by the semi-circle formed by attached solid particles. After 36 hours, opening of the mouth and formation of the gill slits begin. At the age of 90 hours (4 mm.) the larvas swim well and start to feed themselves; the digestive tube is now all open and the swimbladder works already. During the first days of life, larvas have an adhesive organ situated at their frontal region (fig. 7) in form of a crescent, by means of which they hang to surrounding vegetation (fig. 6). When the larva begins to swim and to feed itself and its yolk are having been absorbed. the adhesive organ retracts and disappears. While larvas and alevins feed themselves with plancton, they have small eye-teeth, which disappear,. when fishes become « ilyophagus ». There exist too, during their life as larvas, pharyngeal-teeth. The lateral line appears in the larva after 16 to 18 days; more or less at the same time all fins are completely developed. Shortly after, first scales appear (20 to 23 days). Evolution of intestines twisting followed (fig. 9). Larvas show at different parts of their bodies small of organs excretory functions, that are constituted by bottons in serial disposition, every one with an excretory canal that opens towards the outside. These formations disappear suddenly when larvas attain their phase of alevin. The existence of a great number of said formations at the caudal fin (fig. 12) is of great interest. In our experiences of breeding we have employed several thousands of C. elegans larvas in different environs and we made conditions of surrounding change (illumination), depth of water, temperature, presence of sand at bottom of aquariums and without sand, food). In this way we could compare the results obtained, estimate the action of each factor for the realisation of a good bring-up of larvas.

Orientation of the snail Biomphalaria straminea (Dunker, 1848) in response to light in a situation of selection

The behavioral response of Biomphalaria straminea to light was evaluted in terms of location of the snail in a Y-shaped aquarium in a situation of selection and of the rate (cm/hour) and direction of locomotion under homogeneous 9vertical) or differential (horizontal) lighting upon only one arm of the aquarium. The light source consisted of daylight fluorescent lamps with a spectrum close to that of natural light, with illumination varying from 28 to 350 lux. Analysis of the data showed that all animals, whether in groups or isolated, were attracted to light, although the time needed to approach the light source was 50% shorter for the former than for the latter. The rate of locomotion of B. straminea was 35% higher than observed in B. glabrata and 51% higher than that observed in B. tenagophila studied under similar conditions. The results are discussed in terms of social factors and geographical distribution of the three species.

Daily pattern of flight activity of Aedes albifasciatus in Central Argentina

Aedes albifasciatus is an important common mosquito in Central Argentina. Its a confirmed vector of the Western Equine Encephalitis and is responsible for loss of milk production in dairy cattle during peak populations. This paper reports the flight activity pattern of Ae. albifasciatus for different seasons, in the southern coast of the Mar Chiquita Lake (Central Argentina). Data were collected by sampling two sites between 1992 and 1993 with CDC traps and human-bait captures. Adult mosquito population density, estimated by CDC trapping and human-bait, were highly correlated. However, when compared to other species, the proportion of Ae. albifasciatus was higher in human-bait collections. Adult female populations were active only when temperature were higher than 6§C. Two daily biting peaks were observed (dusk and dawn) during the spring, summer and autumn, and only one peak during winter (around 15:00 pm). Adult abundance was significantly correlated (R²= 0.71; p<0.01) with temperature and illumination.

An ocellar "pupil" that does not change with light intensity, but with the insect age in Triatoma infestans

The simple eyes (ocelli) of recently emerged adult Triatoma infestans exhibit a narrow elongated "pupil", surrounded by a ring of brown-reddish pigment, the "iris". This pupil does not respond to changes in the illumination, but varies in size after the imaginal ecdysis. This change corresponds, internally, with the growth of the corneal lens and the associated retina up to an age of about 20 days. This has not been previously observed in an insect. The use of this characteristic for recognising young adults of this species is suggested.