The use of the polymerase chain reaction and restriction fragment length polymorphism technique associated with the classical morphology for characterization of Lymnaea columella, L. viatrix, and L. diaphana (Mollusca: Lymnaeidae)

The specific identification of Lymnaeid snails is based on a comparison of morphological characters of the shell, radula, renal and reproductive organs. However, the identification is complicated by dissection process, intra and interspecific similarity and variability of morphological characters. In the present study, polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP) techniques targeted to the first and second internal transcribed spacers (ITS1 and ITS2) rDNA and to the mitochondrial 16S ribosomal gene (16S rDNAmt) were used to differentiate the species Lymnaea columella, L. viatrix, and L. diaphana from some localities of Brazil, Argentina, and Uruguay as well as to verify whether the molecular results corroborates the classical morphological method.PCR-RFLP analysis of the ITS1, ITS2, and 16S using 12 restriction enzymes revealed characteristic patterns for L. columella and L. diaphana which were concordant with the classical morphology. On the other hand, for L. viatrix populations a number of 1 to 6 profiles were generated while morphology provided the species pattern results.

Lymnaea cousini Jousseaume, 1887 (Gastropoda: Lymnaeidae): first record for Venezuela

Lymnaea cousini Jousseaume, 1887 was collected in Mucubaji, Merida State, Venezuela, from a permanent pond located at a very high altitude (3760 m). Identification of the collected specimens was made by comparison with the original description of the shell by Jousseaume and the description of the renal organ and reproductive system of topotypic specimens by Paraense.

Detection of Brazilian spotted fever infection by polymerase chain reaction in a patient from the state of São Paulo

Brazilian spotted fever (BSF) cases have been increasing in the state of São Paulo but no genomic information about local rickettsia isolated from humans has been well documented. We recovered spotted-fever group rickettsiae from a sample of patient blood cultured in Vero cells using the shell vial technique. Rickettsial DNA fragments (gltA, ompA, and, ompB genes) were detected, and analysis of the ompB gene base sequences showed identity with the Rickettsia rickettsii ompB sequence available in the GenBank.

Synonymy between Lymnaea bogotensis Pilsbry, 1935 and Lymnaea cousini Jousseaume, 1887 (Gastropoda: Lymnaeidae)

The lymneid snail Lymnaea bogotensis Pilsbry 1935 is synonymized with L. cousini Jousseaume 1887, based on morphological comparisons of the reproductive systems. The shell, renal tube, and reproductive system are described and illustrated from specimens collected in the type locality and the municipality of Paipa, Colombia. Bibliographical records reveal L. columella to be the commonest lymneid in the country. The latter two species can be differentiated based on the morphology of the penial complex.

Comparison between morphological and staining characteristics of live and dead eggs of Schistosoma mansoni

Schistosoma mansoni eggs are classified, according to morphological characteristics, as follows: viable mature and immature eggs; dead mature and immature eggs, shells and granulomas. The scope of this study was to compare the staining characteristics of different morphological types of eggs in the presence of fluorescent labels and vital dyes, aiming at differentiating live and dead eggs. The eggs were obtained from the intestines of infected mice, and put into saline 0.85%. The fluorescent labels were Hoechst 33258 and Acridine Orange + Ethidium Bromide and vital dyes (Trypan Blue 0.4% and Neutral Red 1%). When labelled with the probe Hoechst 33258, some immature eggs, morphologically considered viable, presented fluorescence (a staining characteristic detected only in dead eggs); mature eggs did not present fluorescence, and the other types of dead eggs, morphologically defined, showed fluorescence. As far as Acridine Orange + Ethidium Bromide are concerned, either the eggs considered to be live, or the dead ones, presented staining with green color, and only the hatched and motionless miracidium was stained with an orange color. Trypan Blue was not able to stain the eggs, considered to be dead but only dead miracidia which had emerged out of the shell. Neutral Red stained both live and dead eggs. Only the fluorescent Hoechst 33258 can be considered a useful tool for differentiation between dead and live eggs.

Low susceptibility of Achatina fulica from Brazil to infection with Angiostrongylus costaricensis and A. cantonensis

Introduction of Achatina fulica in Brazil has led to serious concerns about its role as vector for metaIylid worms: AngioIylus costaricensis and A. cantonensis. Experimental infection with both parasites was performed to evaluate the potential risk for their transmission by the giant African snail. Groups of 5 animals, both wild and bred at captivity were exposed at different inocula: 1, 5, and 10 ´ 10³ L1 of A. costaricensis and A. cantonensis. In all groups, few snails got infected and parasitic burden was low. Two different ways of infection were tested: ingestion produced higher numbers of L3 than the inoculation through an artificial hole in the shell. We also report the parasitological examination of 6 batches of wild A. fulica from Florianópolis, state of Santa Catarina, Brazil: only 1 out of 244 animals were infected with metaIylid larvae. Taken together these data indicate that the giant African snail occurring in Southern Brazil is not a permissive host for both AngioIylus species and does not represent a significant risk for transmission of these parasites.

Fine structure of Henneguya hemiodopsis sp. n. (Myxozoa), a parasite of the gills of the Brazilian teleostean fish Hemiodopsis microlepes (Hemiodontidae)

A fish-infecting myxosporean, Henneguya hemiodopsis sp. n., found infecting the gills of Hemiodopsis microlepis and collected from the Poty River near the city of Teresina, Brazil, was described based on ultrastructural studies. The parasite occurred within large whitish polysporic plasmodia (up to 200 μm in diameter) containing asynchronous developmental sporogonic stages, mainly mature spores. The spores measured 19.7 ± 0.9 μm in total length (n = 30) and the ellipsoidal spore body was 10.8 ± 0.5 μm long, 3.3 ± 0.4 μm wide and 2.5 ± 0.5 μm thick. The spores were composed of two equal shell valves adhering together along the straight suture line, with each valve having equal-sized caudal tapering tails measuring 8.7 ± 0.6 μm in length. The spores were surrounded by a thin anastomosed network of microfibrils, more evident on the tails. There were two symmetric elongated bottle-like polar capsules 3.5 ± 0.3 μm long and 1.0 ± 0.2 μm wide, each with a polar filament with five to six coils. Given the morphological and ultrastructural differences from previously described parasites and the specificity of the host species, we propose a new species, named H. hemiodopsis sp. n.

Immunostimulatory property of a synthetic peptide belonging to the soluble ATP diphosphohydro-lase isoform (SmATPDase 2) and immunolocalisation of this protein in the Schistosoma mansoni egg

A peptide (SmB2LJ; r175-194) that belongs to a conserved domain from Schistosoma mansoni SmATPDase 2 and is shared with potato apyrase, as predicted by in silico analysis as antigenic, was synthesised and its immunostimulatory property was analysed. When inoculated in BALB/c mice, this peptide induced high levels of SmB2LJ-specific IgG1 and IgG2a subtypes, as detected by enzyme linked immunosorbent assay. In addition, dot blots were found to be positive for immune sera against potato apyrase and SmB2LJ. These results suggest that the conserved domain r175-194 from the S. mansoni SmATPDase 2 is antigenic. Western blots were performed and the anti-SmB2LJ antibody recognised in adult worm (soluble worm antigen preparation) or soluble egg antigen antigenic preparations two bands of approximately 63 and 55 kDa, molecular masses similar to those predicted for adult worm SmATPDase 2. This finding strongly suggests the expression of this same isoform in S. mansoni eggs. To assess localisation of SmATPDase 2, confocal fluorescence microscopy was performed using cryostat sections of infected mouse liver and polyclonal antiserum against SmB2LJ. Positive reactions were identified on the external surface from the miracidium in von Lichtenberg's envelope and, in the outer side of the egg-shell, showing that this soluble isoform is secreted from the S. mansoni eggs.

Ultrastructural description of Ceratomyxa microlepis sp. nov. (Phylum Myxozoa): a parasite infecting the gall bladder of Hemiodus microlepis, a freshwater teleost from the Amazon River

A new ceratomyxid parasite was examined for taxonomic identification, upon being found infecting the gall bladder of Hemiodus microlepis (Teleostei: Hemiodontidae), a freshwater teleost collected from the Amazon River, Brazil. Light and transmission electron microscopy revealed elongated crescent-shaped spores constituted by two asymmetrical shell valves united along a straight sutural line, each possessing a lateral projection. The spores body measured 5.2 ± 0.4 µm (n = 25) in length and 35.5 ± 0.9 µm (n = 25) in total thickness. The lateral projections were asymmetric, one measuring 18.1 ± 0.5 µm (n = 25) in thickness and the other measuring 17.5 ± 0.5 µm (n = 25) in thickness. Two equal-sized subspherical polar capsules measuring 2.2 ± 0.3 µm in diameter were located at the same level, each possessing a polar filament with 5-6 coils. The sporoplasm was binucleate. Considering the morphometric data analyzed from the microscopic observations, as well as the host species and its geographical location, this paper describes a new myxosporean species, herein named Ceratomyxa microlepis sp. nov.; therefore representing the first description of a freshwater ceratomyxid from the South American region.

On the immature stages and natural history of Gonioterma exquisita Duckworth (Lepidoptera, Elachistidae, Stenomatinae)

In the present work the natural history and immature stages of Gonioterma exquisita Duckworth, 1964 of the Brazilian cerrado region are described. The caterpillars are external folivorous feeders and present a diet restricted to pubescent-leaved host plants of the genus Byrsonima Rich. (Malpighiaceae). They are solitary caterpillars and they build a hard individual shelter that looks like a planorbid shell, made from silk and frass, covered with trichomes and silk, where they develop until the last larval instar on the host plant, and remain inside it until the adult emergence on herbaceous stratum. The larval development presented eight instars. Egg, larval head capsules, instar duration, pupa, shelter, and behavior are also described. Diapause is mentioned for the first time in this species.

Parboiled rice whole bran in laying diets for Japanese quails

The objective of this work was to evaluate the effects of parboiled rice whole bran (PRWB) inclusion in laying diets for Japanese quails, on their performance, egg quality, and economic viability. A total of 448 17-week-old quails were weighed and distributed in a completely randomized design with seven treatments and eight replicates of eight birds each. A control diet (no PRWB) and six diets, containing 5, 10, 15, 20, 25, and 30% of PRWB, were tested. The increasing levels of PRWB did not affected nutrient digestibility coefficient, dietary energy use, feed intake, egg production, egg weight, egg mass, and the economic viability indices. However, there was a linear decrease in egg shell percentage, specific weight, and yolk color. The inclusion of up to 30% PRWB in the diet allows nutrient utilization and performance similar to those obtained by the control group, and it is economically viable.

Growth of Crassostrea gasar cultured in marine and estuary environments in Brazilian waters

The objective of this work was to evaluate the growth of the mangrove oyster Crassostrea gasar cultured in marine and estuarine environments. Oysters were cultured for 11 months in a longline system in two study sites - São Francisco do Sul and Florianópolis -, in the state of Santa Catarina, Southern Brazil. Water chlorophyll-α concentration, temperature, and salinity were measured weekly. The oysters were measured monthly (shell size and weight gain) to assess growth. At the end of the culture period, the average wet flesh weight, dry flesh weight, and shell weight were determined, as well as the distribution of oysters per size class. Six nonlinear models (logistic, exponential, Gompertz, Brody, Richards, and Von Bertalanffy) were adjusted to the oyster growth data set. Final mean shell sizes were higher in São Francisco do Sul than in Florianópolis. In addition, oysters cultured in São Francisco do Sul were more uniformly distributed in the four size classes than those cultured in Florianópolis. The highest average values of wet flesh weight and shell weight were observed in São Francisco do Sul, whereas dry flesh weight did not differ between the sites. The estuary environment is more promising for the cultivation of oysters.

Limestone particle sizes and lighting regimens on egg and bone quality of laying hens

Abstract:The objective of this work was to evaluate the effect of limestone particle sizes in the diet and of lighting regimes on the egg and bone quality and on the performance of commercial laying hens. Three hundred Hissex White layers, at 18 weeks of age, were distributed in a completely randomized design, in a 5×2 factorial arrangement (coarse limestone in the diet at 0, 25, 50, 75, and 100%; with or without artificial light), with five replicates of six birds. No significant interaction was observed between particle sizes and lighting regime for the evaluated parameters. There was no significant effect of coarse limestone level in the diet on the performance and egg quality of hens; however, bone deformity (3.23 to 4.01 mm), strength (5.19 to 6.70 kgf cm-2), and mineral matter (51.09 to 59.61%) improved as the proportion of coarse limestone increased. For lighting regime, the treatment with artificial light yielded higher Haugh unit values (87.17 vs. 85.54) than that with natural light only. Greater limestone particles improve bone quality of laying hens, and the use of artificial light can benefit the albumen quality of the eggs.

Peduncle and fruit yield, in six cropping seasons, of early dwarf cashew tree clones irrigated with different water regimes

The objective of this work was to evaluate peduncle and fruit yield in clone MS 076 and in a clonal population of drip-irrigated, early dwarf cashew trees propagated by layering, in six cropping seasons. In order to meet the increased water requirements of the crop resulting from plant growth and development, irrigation during the dry season was performed daily according to the following water regime: 15 min/plant/day during the 1st year, 30 min/plant/day during the 2nd year, 45 min/plant/day during the 3rd year and 60 min/plant/day during all subsequent years. Water was supplied by one drip emitter/plant, at an (adjustable) flow rate of 36 L/h.The research was carried out in Fortaleza-Ceará, Brazil, and a random block design was utilized, with five replicates and split-plots. The clones were assigned to plots and the cropping seasons were considered as subplots. The clonal population was superior to the clone only with regard to number of nut shells (NNS), and solely in the first season. The clone was superior to the population as to NNS and peduncle yield (PY) in the second season, and also with regard to the three evaluated traits - NNS, PY, and nut shell yield, in the last three cropping seasons.

Development of a shelling method to recover whole kernels of the Cutia nut (Couepia edulis)

The kernel of the cutia nut (castanha-de-cutia, Couepia edulis (Prance) Prance) of the western Amazon, which is consumed by the local population, has traditionally been extracted from the nut with a machete, a dangerous procedure that only produces kernels cut in half. A shelling off machine prototype, which produces whole kernels without serious risks to its operator, is described and tested. The machine makes a circular cut in the central part of the fruit shell, perpendicular to its main axis. Three ways of conditioning the fruits before cutting were compared: (1) control; (2) oven drying immediately prior to cutting; (3) oven drying, followed by a 24-hour interval before cutting. The time needed to extract and separate the kernel from the endocarp and testa was measured. Treatment 3 produced the highest output: 63 kernels per hour, the highest percentage of whole kernels (90%), and the best kernel taste. Kernel extraction with treatment 3 required 50% less time than treatment 1, while treatment 2 needed 38% less time than treatment 1. The proportion of kernels attached to the testa was 93%, 47%, and 8% for treatments 1, 2, and 3, respectively, and was the main reason for extraction time differences.