Habitat use by a tree frog species of Scinax (Amphibia, Hylidae) at an urban forest fragment from south-eastern Brazil

We studied the pattern of habitat use by the tree frog Scinax aff. perereca. Fieldworks were performed from August 1996 to August 1997 at Parque das Mangabeiras, Belo Horizonte, State of Minas Gerais, southeastern Brazil. Calling males were observed in September, October and December 1996, February to April and June 1997. Females were found only in October 1996. Specimens were found perched on vegetation, on the ground or on stones near waterfall. At Parque das Mangabeiras, S. aff. perereca occupied nine types of substrata. The most frequently used substrata were shrubs, stones at the stream edges, and fallen trunks. The pattern of spatial occupation varied among months. Males were found calling in aggregations on the vegetation and spatial niche breadth was related to species abundance.

Feeding ecology of Pterodoras granulosus (Siluriformes, Doradidae) in the Lajeado Reservoir, Tocantins, Brazil

The diet and feeding habits of armado catfish, Pterodoras granulosus (Valenciennes, 1821), were studied in the Lajeado Reservoir, Tocantins, Brazil, and the mouth of its tributaries. Stomach contents of 327 specimens were analyzed by the percentage composition and volumetric methods. The feeding of armado on a wide variety of foods, including both animal and plant items, leads to its classification as a euriphagic species with herbivorous tendency. However, it should be noted that armado showed a strong ontogenetic diet shift, with the gradual replacement of detritus and sediment by plant items, especially terrestrial ones. The environmental use pattern of P. granulosus corroborates the ontogenetic shift observed in its diet. Small fish predominated mainly in the benthic region and detritus and sediment represented an important resource, whereas large fish fed mainly on terrestrial plants, as they explore open water sites and the surface of the water column.

Seasonal dynamics of a drosophilid (Diptera) assemblage and its potencial as bioindicator in open environments

Drosophila Fallen, 1823 (Diptera, Drosophilidae) is for long a well-established model organism for genetics and evolutionary research. The ecology of these flies, however, has only recently been better studied. Recent papers show that Drosophila assemblies can be used as bioindicators of forested environment degradation. In this work the bioindicator potential of drosophilids was evaluated in a naturally opened environment, a coastal strand-forest (restinga). Data from nine consecutive seasonal collections revealed strong temporal fluctuation pattern of the majority of Drosophila species groups. Drosophila willistoni group was more abundant at autumns, whereas D. cardini and D. tripunctata groups were, respectively, expressive at winters and springs, and D. repleta group at both seasons. The exotic species D. simulans Sturtevant, 1919 (from D. melanogaster group) and Zaprionus indianus Gupta, 1970 were most abundant at summers. Overall, the assemblage structure did not show the same characteristics of forested or urban environments, but was similar to the forests at winters and to cities at summers. This raises the question that this locality may already been under urbanization impact. Also, this can be interpreted as an easily invaded site for exotic species, what might lead to biotic homogenization and therefore can put in check the usage of drosophilid assemblages as bioindicators at open environments.

Reproduction and feeding ecology of Amphisbaena munoai and Anops kingi (Amphisbaenia, Amphisbaenidae) in the Escudo Sul-Rio-Grandense, southern Brazil

This study presents original data on the reproduction and feeding ecology of two syntopic amphisbaenians, Amphisbaena munoai Klappenbach, 1969 and Anops kingi Bell, 1833, from southern Brazil. Sampling was carried out from April 2004 to April 2006 at a highland area located in São Jerônimo and Barão do Triunfo municipalities, Rio Grande do Sul, Brazil. A total of 647 amphisbaenians were collected, 510 specimens of A. munoai and 137 specimens of A. kingi, of which 130 and 93, respectively, had their gonads and gut content analyzed. Both species presented a unimodal pattern of seasonal distribution, sexually mature females with significantly larger snout vent length (SVL) than sexually mature males, and seasonal reproductive cycle. Adults of A. kingi had significantly larger (SVL) than those of Amphisbaena munoai. Both species had generalist diets, but termites (Isoptera) was a staple item in their diet. Greater predominance of insect larvae was observed in the diet of A. kingi, which may be due to its larger body in comparison to A. munoai. Insect larvae ingested by A. kingi were significantly larger than those ingested by A. munoai. Diets of adult males and females of both species were not significantly different. The ontogenetic diet comparison in both species revealed significant numerical differences, with adults consuming higher numbers of prey.

Description of the male of Mangora brokopondo (Araneae, Araneidae), with notes on Mangora species from Brazilian Oriental Amazon

The male of Mangora brokopondo Levi, 2007 is described and illustrated for the first time. Variation in the color pattern of the females is documented. Mangora woytkowskii Levi, 2007 is considered a junior synonym of M. hirtipes (Taczanowski, 1878). New records of M. alinahui Levi, 2007 and M. pia Chamberlin & Ivie, 1936 are presented.

The influence of temperature and humidity on abundance and richness of Calliphoridae (Diptera)

The blowfly species are important components in necrophagous communities of the Neotropics. Besides being involved in the degradation of animal organic matter, they may serve as vectors for pathogens and parasites, and also cause primary and secondary myiasis. The occurrence pattern of these species is well defined, yet it is still not very clear which of these environmental factors determine the structure of the assemblies. This paper was developed to evaluate the influence of mean temperature and relative humidity variation in the abundance and richness of blowflies in the Brazilian southernmost state, Rio Grande do Sul, where temperature variation is well marked throughout the year. To evaluate this objective, WOT (Wind Oriented Trap) were installed with beef liver as bait in three environments for 10 consecutive days in each month between July 2003 and June 2004. A total of 13,860 flies were collected distributed among 16 species with a higher frequency of Lucilia eximia (Wiedemann, 1819) and Chrysomya albiceps (Wiedemann, 1819). The mean temperature and relative humidity influence the richness of blowflies, with greater richness and abundance in late spring and early summer, whereas abundance was only influenced by temperature. Each species responded differently with respect to these variables, where L. eximia is not influenced by any of the two abiotic factors, despite the high abundance presented. This paper presents the results of the sensitivity for the presence or absence of species of Calliphoridae and on the variation of the abundance of these species under regime temperature changes and relative humidity with implications for public health and animal management.

A new species of Leiostracus from Bahia, Brazil (Gastropoda, Pulmonata, Orthalicidae)

A remarkable new species of pulmonate snail was recently collected in a small Atlantic Rainforest fragment near the city of Canavieiras, state of Bahia, Brazil, an area known for a high diversity of land snails. It is described herein as Leiostracus fetidus sp. nov. and can be easily identified by its color pattern of irregular brown to black axial stripes on a white to yellow background, a reddish axial band "separating" the white peristome from the rest of the shell and a broad brown spiral band surrounding the umbilical region. Other diagnostic features include a relatively small size, a proto columellar fold and two very weak folds delimiting the basal region of the aperture. This discovery is a reminder of how little this fauna is known and also an alarm for proper conservation of these forest fragments.

Reproductive strategy of a non-annual rivulid in a perennial wetland

ABSTRACT Atlantirivulus riograndensis (Costa & Lanés, 2009) is a fish registered to the basin of Patos lagoon and the adjacent coastal plains in southern states of Rio Grande do Sul and Santa Catarina, Brazil, found in shallow water courses with that have large quantities of aquatic vegetation and forest edges. The objective of this study was to investigate the reproductive strategy and tactics of this species including the sex ratio, the length at first maturity, spawning type, fecundity and the possible associations among reproduction and abiotic factors. Sampling of specimens occurred in perennial wetlands within the Banhado dos Pachecos wildlife refuge, in the city of Viamão, state of Rio Grande do Sul, Brazil, which is a conservation unit that belongs to an area of environmental protection in the Pampa Biome. The capture of 30 A. riograndensis specimens per month occurred from January to December of 2012. A total of 188 females and 172 males were captured and the total sex ratio was 1:1 in the sampled population. Sexual maturity of the species occurs after 13.59 and 11.92 mm (SL) for females and males, respectively. Both a multiple spawning and a long reproductive period (since August to March) were confirmed by the presence of post-ovulatory follicles that were observed through histological analysis and the values of the gonadosomatic index in females considered spawning capable. The average absolute fecundity of the species is of 19.33 (± 6.18) vitellogenic oocytes in mature ovaries. No significant relationship was found between mean GSI and the abiotic data. Reproductive tactics presented by A. riograndensis indicate a species with an opportunistic reproductive strategy, following the pattern of other species of the Rivulidae family.

The interaction of Gram negative bacteria and S. mansoni in mice with experimental Schistosomiasis

Animals (122 mice) were infected each with eighty cercariae of S. mansoni and subsequently challenged intravenously eight weeks later with the following gram-negative organisms. S. typhi, E. coli, Klebsiella-enterobacter species, Proteus mirabilis and Pseudomonas aeruginosa. Enumeration of bacteria in the liver, spleen and blood and S. mansoni from the portal sistem was performed from one to four weeks later in infected animals. A significant difference between infection produced by S. typhi and other gram negative organisms was observed: S. typhi persisted longer in the spleen and liver and could be recovered from S. mansoni worms up to three weeks following bacterial infection. Other gram negative bacteria disappeared from S. mansoni worms after two weeks of initial challenge. Additional animals (51 mice) infected with S. mansoni were given S. typhi, E. coli or sterile saline. After two weeks, animals were sacrificed and the recovery rate of worms from the portal system, and the mesenteric and hepatic oogram were determined. in animals infected with E. coli a significant decrease in the number of worms was observed compared to the saline control group; thirty worms were recovered in the control group compared to two worms in e. coli infected animals. In addition, the patterns of oviposition was significantly different in these latter animals suggesting complete inhibition of this process. Following S. typhi infection the difference in recovery of worms and pattern of oviposition was minimal. These findings suggest a difference in the interaction of various gram negative bacteria and S. mansoni and are consistent with the clinical observation of prolonged salmonella bacteremia in patients with schistosomiasis.

A simple method to purify biologically and antigenically preserved bloodstream trypomastigotes of Trypanosoma cruzi using Deae-cellulose columns

A method to purify trypanosomastigotes of some strains of Trypanosoma cruzi (Y, CL, FL, F, "Berenice", "Colombiana" and "São Felipe") from mouse blood by using DEAE-cellulose columns was standardized. This procedure is a modification of the Lanham & Godfrey methods and differs in some aspects from others described to purify T. cruzi bloodstream trypomastigotes, mainly by avoidance of prior purifications of parasites. By this method, the broad trypomastigotes were mainly isolated, accounting for higher recoveries obtained with strains having higher percentages of these forms: processing of infected blood from irradiated mice could be advantageous by increasing the recovery of parasites (percentage and/or total number) and elution of more slender trypomastigotes. Trypomastigotes purified by this method presented normal morphology and motility, remained infective to triatomine bugs and mice, showing in the latter prepatent periods and courses parasitemia similar to those of control parasites, and also reproducing the polymorphism pattern of each strain. Their virulence and pathogenicity also remained considerably preserved, the latter property being evaluated by LD 50 tests, mortality rates and mean survival time of inoculated mice. Moreover, these parasites presented positive, clear and peripheral immunofluorescence reaction at titres similar to those of control organisms, thus suggesting important preservation of their surface antigens.

Effect of drugs on Trypanosoma cruzi and on its interaction with heart muscle cell in vitro

Megazol, nifurtimox, benznidazol and allopurinol were investigated, by light and electron µscopy, for their action on T. cruzi. Both the direct effect upon amastigote and trypomastigote forms and the effect upon the interaction of heart muscle cells (HMC) with bloodstream trypomastigotes were studied. The proliferation of amastigotes in Warren medium was inhibited in a dose-dependent manner by megazol, nifurtimox and benznidazol. Treatment of amastigotes (25-50 µM/24 h) and trypomastigotes (25 µM/24h) led to several ultrastructural alterations in the parasites. These three drugs also had a potent effect on the treatment of infected heart muscle cells when added at the beginning of the interaction or after one or three days of infection. The interiorized parasites showed a similar pattern of ultrastructural alterations as observed by the direct effect on the amastigotes. The primary heart muscle cell culture proved to be a suitable model for the study of drugs on intracellular parasites. Likewise, the amastigote proliferation in axenic medium was shown to be an adequate assay for an initial trial of drugs. These parameters seem very reliable to us for a systematic investigation of the mechanism of action of new drugs.

Extra-tissular Schistosoma mansoni egg granulomata in the peritoneal cavity of mice

The presence of Schistosoma mansoni eggs surrounded by inflammatory cells were detected within the peritoneal cavity of experimentally infected mice. The histological and ultrastructural analysis revealed the predominantly macrophagic composition of these structures. The presence of epithelioid cells, macrophages in different stages of activation and the architectural pattern of the cells, characterize these structures as extra-tissular true granulomas. Granulomas much similar to those observed in the peritoneal cavity of infected mice were also detected after the intraperitoneal injection of viable eggs in non-infected mice. Collagen fibers were observed in between the inflammatory cells of granulomas obtained 10 weeks after infection and 48 hours after the injection of viable eggs into the peritoneal cavity. In later times of infection or injection the amount of collagen fibers increases resulting in a typical pattern of healed schistosoma egg granulomas. The possible influence of the immune response on the genesis of the granulomatous reaction as well as the influence of the vascularized connective tissue on this process is discussed.

The thermal stability of yellow fever vaccines

The assessment of yellow fever vaccine thermostability both in lyophilized form and after reconstitution were analyzed. Two commercial yellow fever vaccines were assayed for their thermal stability. Vaccines were exposed to test temperatures in the range of 8 (graus) C to 45 (graus) C. Residual infectivity was measured by a plaque assay using Vero cells. The titre values were used in an accelerated degradation test that follows the Arrhenius equation and the minimum immunizing dose was assumed to be 10 (ao cubo) particles forming unit (pfu)/dose. Some of the most relevant results include that (i) regular culture medium show the same degradation pattern of a reconstituted 17D-204 vaccine; (ii) reconstituted YF-17D-204 showed a predictable half life of more than six days if kept at 0 (graus) C; (iii) there are differences in thermostability between different products that are probably due to both presence of stabilizers in the preparation and the modernization in the vaccine production; (iv) it is important to establish a proper correlation between the mouse infectivity test and the plaque assay since the last appears to be more simple, economical, and practical for small laboratories to assess the potency of the vaccine, and (v) the accelerated degradation test appears to be the best procedure to quantify the thermostability of biological products.

Reversibility of cardiac fibrosis in mice chronically infected with Trypanosoma cruzi, under specific chemotherapy

This investigation was performed to verify the effect of specific chemotherapy (Benznidazole or MK-346) on the inflammatory and fibrotic cardiac alterations in mice chronically infected with the strains 21 SF (Type II) and Colombian (Type III) of Trypanosoma cruzi. To obtain chronically infected mice, two groups of 100 Swiss mice each, were infected with either the 21 SF or the Colombian strain (2x 10 [raised to the power of] 4 and 5x 10 [raised to the power of] 4 blood forms respectively). The rate of morality in the acute phase was of 80% for both groups. Twenty surviving mice chronically infected with the 21 SF strain and 20 with the Colombian strain were then divided in treated and untreated groups. Excluding those that died during the course of treatment, 14 mice chronically infected with the 21 SF strain and 15 with the Colombian strain were evaluated in the present study. Chemotherapy was performed with Benznidazole (N-benzil-2-nitro-1-imidazolacetamide) in the dose of 100mg/k.b.w/day, for 60 days, or with the MK-436(3(1-methyl-5 nitroimidazol-2-yl) in two daily doses of 250 mg/k.b.w, for 20 days. Parasitological cure tests were performed (xenodiagnosis, haemoculture, subinovulation of the blood into newborn mice), and serological indirect immunofluorescence test. The treated and untreated mice as well as intact controls were killed at different periods after treatment and the heart were submitted to histopathological study with hematoxilineosin and picrosirius staining; ultrastructural study; collagen immunotyping, fibronectin and laminin identification by immunofluorescence tests. Results: the untreated controls either infected with 21 SF or Colombian strain, showed inflammatory and fibrotic alterations that were mild to moderate with the 21 SF strain and intense with the Colombian strain. Redpicrosirius staining showed bundles of collagen in the interstitial space and around cardiac fibers. Increased deposits of mitritial components and collagen fibers, macrophages and fibroblasts appeared at the ultra structural examination. Deposits of fibronectin, laminin, pro-III and IV collagens were seen, most intense in those infected with the Colombian strain. Treated nice, parasitologically cured, presented clear-cut regression of the inflammatory lesions and of the interstitial matrix thickening. Mice infected with the Colombian strain and treated with MK-436, was parasitologically cured in 5/6 cases and showed mild inflammatory infiltration and fibrosis. The mice treated with Benznidazole (Colombian strain) did not cure and showed moderate fibrosis and inflammation. Treatment of the nice infected with the 21 SF with Benznidazole determined parasitological cure of all animals, that showed mild inflammation and fibrosis of the myocardium. The cured mice of all groups and treated but uncured showed collagen degradation at electronmicroscopy and decrease of immunofluorescence pattern of the matrix.

Dynamics of fibrosis production and resorption in intestinal schistosomiasis of mice

A histological, morphometric and immunocytochemical study of schistosomal periovular granulomas in the liver and intestines of mice revealed that intestinal granulomas are smaller and contain less collagen than those in the liver. After curative treatment intestinal granulomas undergo a relatively more rapid resorption, although the general pattern of collagen degradation apparently does not differ from that observed in the liver. Tendency to form scattered, usually isolated granulomas that are only mildly fibrogenic, coupled with a well-balanced process of resorption appear as the explanation why intestinal fibrosis is not an outstanding feature of schistosomiasis as it is in the liver.