Hatching and larval export of the intertidal crab Neohelice granulata in Mar Chiquita coastal lagoon, Argentina

Synchronization in the events of the reproductive cycle in female Neohelice granulata Dana, 1851 were studied from samples taken weekly and biweekly from September to December 2006 in the Laguna Mar Chiquita. The timing and larval hatching and synchronicity were inferred from numbers of ovigerous females and observing the stages of embryonic development. Synchronization in larval hatching also was observed in females in experiments in dark for a period of 48 hours, at three different salinities (10, 23 and 33 ppm). In addition plankton sampling were performed in order to study larval exportation at the field and its link to the tidal and light/dark cycles. We found that ovigerous females of N. granulata have a marked synchronization in embryonic development which results in that most of berried females are close to hatching within a period of maximum tidal range (days). Within this period, there is a synchronization of hatching at a time scale of hours, governed by environmental conditions. The salinity range used in this study (10-32‰) did not affect hatching synchronicity neither time to hatch. Hatching was synchronized according to endogenous rhythms governed mainly by the tidal cycle and secondarily by the breadth of it. It is also conditioned by the light-dark cycle through an exogenous cycle, so that the hatchings would occur mostly at night high tides.

Variation in the hatching response of Ochlerotatus albifasciatus egg batches (Diptera: Culicidae) in temperate Argentina

Egg hatching of winter-collected Ochlerotatus albifasciatus was studied for six months. Batches of eggs were divided into two groups, one of them was stored in the laboratory at 23°C and 12:12 photoperiod, and the other in the field under dead leaves. Every month, from July to December, eggs from the two groups were flooded under both laboratory and field conditions. Unhatched eggs were returned to the original condition and flooded two more times separated by ten-day intervals. Results show that egg diapause is expressed in different intensities, not only on eggs exposed to different conditions but also in those exposed to the same condition, even when they were laid by the same female. Successive inundations yielded incomplete hatches of eggs, and favored the hatching response in the next flooding. Low environmental temperatures before and during the flooding depressed hatching response. This shows that eggs need a warm period before flooding as well as warm temperatures during flooding, to hatch. As drought period was longer hatching response increased, but this was also accompanied by warmer environmental conditions. The experiment performed in laboratory did not show that increment. Field studies showed that a layer of dead leaves protected eggs from extreme temperatures.

Latex constituents from Calotropis procera (R. Br.) display toxicity upon egg hatching and larvae of Aedes aegypti (Linn.)

Calotropis procera R. Br. (Asclepiadaceae) is a well-known medicinal plant with leaves, roots, and bark being exploited by popular medicine to fight many human and animal diseases. This work deals with the fractionation of the crude latex produced by the green parts of the plant and aims to evaluate its toxic effects upon egg hatching and larval development of Aedes aegypti. The whole latex was shown to cause 100% mortality of 3rd instars within 5 min. It was fractionated into water-soluble dialyzable (DF) and non-dialyzable (NDF) rubber-free materials. Both fractions were partially effective to prevent egg hatching and most of individuals growing under experimental conditions died before reaching 2nd instars or stayed in 1st instars. Besides, the fractions were very toxic to 3rd instars causing 100% mortality within 24 h. When both fractions were submitted to heat-treatment the toxic effects were diminished considerably suggesting low thermostability of the toxic compounds. Polyacrylamide gel electrophoresis of both fractions and their newly fractionated peaks obtained through ion exchange chromatography or desalting attested the presence of proteins in both materials. When submitted to protease digestion prior to larvicidal assays NDF lost most of its toxicity but DF was still strongly active. It may be possible that the highly toxic effects of the whole latex from C. procera upon egg hatching and larvae development should be at least in part due to its protein content found in NDF. However the toxicity seems also to involve non protein molecules present in DF.

A new miracidia hatching device for diagnosing schistosomiasis

It is still imperative to develop a parasitological technique highly sensitive for diagnosing schistosomiasis in epidemiological and individual surveys. A simple and cheap hatching device with a collecting container was manufactured and tested under experimental conditions. Twelve Kato-Katz slides were performed as golden standard for comparison. Quantitative results can be carried out by counting miracidia in a plate and parasite load can be calculated (miracidia/gram of feces). Statistically significant values were higher in the hatching test. More sensitive results, with statistical significance, were achieved using 1.5 g of feces (which corresponds to 36 Kato-Katz slides) than by using the Kato-Katz method. Advantages of this technique and its limitations are presented.

Estimating the most important criteria for hatching eggs as function of broiler breeders age

This research aimed to compare two female broiler breeder ages during the incubation period regarding management using the Analytic Hierarchy Process method (AHP). This method is characterized by the possibility of analyzing a multicriteria problem and assists a decision making. This study was carried out on a commercial hatchery located in São Paulo, Brazil. Two ages of broiler breeder (42 and 56 weeks) were compared relative to production rate. Production index data were the same in both ages and were submitted to multicriteria decision analysis using the AHP method. The results indicate that broiler breeders of 42 weeks presented better performance than those of 56 week-old. The setter phase (incubation) is more critical than the hatcher. The AHP method was efficient for this analysis and can serve as a methodological basis for future studies to improve the hatchability of broilers eggs.

Relationship between weight, age and hatching success and the concentration of heavy metals in nestling blue macaw (Anodorhynchus hyacinthinus Latham, 1790) in the Pantanal,Mato Grosso do Sul

Abstract: The concentration of heavy metals (Cr, Fe, Al, As, Cd, Cu, Pb, Mo, Ni, Se and Zn) was evaluated in the blood of nestling blue macaws (Anodorhynchus hyacinthinus) captured in the Pantanal, Mato Grosso do Sul (n=26) in 2012; this was based on the hypothesis that these birds exhibit levels of these heavy metals in their organism and that these interfere in hatching success, weight and age of the chicks. Blood samples were digested with nitric acid and hydrochloric acid and the quantification of metals was performed by ICP-OES (Optical Emission Spectroscopy and Inductively Coupled Plasma). Blood samples of nestlings showed concentrations of Cr (0.10μg/g) Fe (3.06μg/g) Al (3.46μg/g), Cd (0.25μg/g) Cu (0.74μg/g), Mo (0.33μg/g), Ni (0.61μg/g), Se (0.98μg/g), and Zn (2.08μg/g). The levels of heavy metals found were not associated with weight, age and hatching success of the chicks.

Egg size, yolk mass extrusion and hatching behavior in two cryptic speciesof Anastrepha fraterculus (Wiedemann) (Diptera, Tephritidae)

Variations in egg length were observed for two populations of cryptic species of Anastrepha fraterculus (Wiedemann). The eggs of type I flies were smaller than those of type II individuals. For both types, in regard to yolk mass extrusion, four classes of embryos were detected. Class 1: embryos that extrude masses at both extremities; class 2: embryos in which extrusion occurs only at the anterior pole; class 3: embryos that eliminate mass only at the posterior pole, and class 4: embryos that do not extrude any mass. Embryo class frequencies were similar for populations belonging to the same type, but different between types. Individual females may produce eggs from different embryo classes, but for any given female the pattern remains constant during a long period of oviposition. Variation in size of the extruded masses was similar for both populations. Individual females produced embryos with a small range of mass diameters, and different females produced masses of different mean size. However, individual mass size remained constant during oviposition. The results suggest the existence of genetic components involved in the control of this unusual process. Larvae of both types presented, just before eclosion, similar unusual behaviors: they ingest the anterior extruded mass, rotate 180°, absorb the posterior mass and eclose near the posterior pole. Data show that cryptic A. fraterculus type I and type II differs in regard to egg size as well as to the phenomenon of yolk mass extrusion

Modulation of murine blastocyst hatching in vitro by glutamine and tryptophan

Enrichment of culture media with amino acids improves embryo development. However, little is known about the specific action of each amino acid during embryogenesis. The present study was undertaken to examine the effect of L-glutamine (Gln) and tryptophan (Trp) on mouse embryo hatching, expansion and viability in vitro. Blastocysts were collected from 6- to 8-week-old female BALB/c mice (N = 30) and cultured in M2 medium containing either 0.125, 0.25 or 0.5 mM Trp, 1 mM Gln, or M2 alone. Gln significantly increased (100%; P < 0.05) blastocyst hatching at 24 h compared to M2 alone or Trp; moreover, Trp inhibited blastocyst hatching when compared to M2 alone (P < 0.05) at 72 h. In contrast, the percentage of embryos reaching the state of expanded blastocyst at 48 h was significantly higher in medium with 1 mM Gln (66.6%; P < 0.05) or with 0.125 mM Trp (61.1%; P < 0.05). Unexpectedly, Trp increased the percentage of degenerated blastocysts after 48 h (67.7%; P < 0.05), while Gln preserved blastocyst viability. These results suggest that Gln may enhance blastocyst hatching, expansion and viability in vitro.

Duration of larval and pupal development stages of Aedes albopictus in natural and artificial containers

Aedes albopictus were reared in different containers: a tree hole, a bamboo stump and an auto tire. The total times from egg hatching to adult emergence were of 19.6,27.3 and 37.5 days, respectively, according to the container. The first, second and third-instar larvae presented growth periods with highly similar durations. The fourth-instar larvae was longer than the others stages. The pupation time was longer than the fourth-instar larvae growth period. The temperature of the breeding sites studied, which was of 18° C to 22° C on average, was also taken into consideration. The mortality of the immature stages was analysed and compared as between the experimental groups; it was lower in the natural containers than in the discarded tire. The average wing length of adult females emerging from tree hole was significantly larger (p < 0.05) than that of those emerging from the tire.

Strongyloses ratti and S. stercoralis: effects of cambendazole, thiabendazole and mebendazole in vitro

The effects of in vitro incubation of three henzimidazole anthelmintics, thiabendazole, mebendazole and cambendazole on Strongyloides were compared. No drug affected hatching of S. ratti eggs or the viability of infective larvae or parasitic adult worms, but all three inhibited moulting of S. ratti larvae. In addition, cambendazole, but not thiabendazole or mebendazole, impaired the viability of S. ratti first- and second-stage larvae. The three drugs had no effect on isolated S. stercorais free-living adult worms, but they all prevented development of S. stercoralis rhabditiform larvae. Thiabendazole and mebendazole had no effect on the infectivity of either S. ratti or S. stercoralis infective larvae, but infection with these worms was abrogated by prior incubation with cambendazole. These results indicate that cambendazole acts in a different manner to the other two drugs. Since it is active against larvae migrating through the tissues, it is potentially of much greater value than thiabendazole or mebendazole in the therapy of strongyloidiasis.

Embryology of Triatoma infestans (KLUG), (Hemiptera, Reduviidae), a Chagas' disease vector

This study reports the embryogenesis of T. infestans (Hemiptera, Reduviidae). Morphological parameters of growth sequences from oviposition until hatching (12-14 d 28ºC) were established. Five periods, as percent of time of development (TD), were characterized from oviposition until hatching. The most important morphological features were: 1) formation of blastoderm within 7% of TD; 2) germ band and gastrulation within 30% of TD; 3) nerve cord, limb budding, thoracic and abdominal segmentation and formation of body cavity within 50% of TD; 4) nervous system and blastokinesis end, and development of embryonic cuticle within 65% of TD; 5) differentiation of the mouth parts, fat body, and Malphigian tubules during final stage and completion of embryo at day 12 to day 14 around hatching. These signals were chosen as appropriate morphological parameters which should enable the evaluation of embryologic modifications due to the action/s of different insecticides

Evaluation in white mice of the infectivity of egs of Lagochilascaris minor (Nematoda: ascarididae), incubated by decortication with sodium hypochlorite (NaOCl)

White mice were used to study the infectivity of the eggs of Lagochilascaris minor Leiper, 1909 after incubation in liquid media, with or without preservative substances. Potassium bichromate (K2Cr2O7) at 1% restrict hatching, while 1% formalin gave a greater larval yield. Incubation of eggs in distilled water, in Roux or Falcon flasks gave a good yield, whether the eggs were obtained from human feces or from experimentally infected cats. Treatment of eggs with Sodium hypochlorite (NaOCl) at 5.25% for 2 min prior to inoculation, produced a notable increment of the larval yield in the infections.

Comparative study of the fecundity and fertility of Biomphalaria glabrata (Say, 1818) and Biomphalaria straminea (Dunker, 1848) in a laboratory through self-fertilization and cross-fertilization

The aim of this study was to compare the fecundity and fertility of B. glabrata and B. straminea by cross- and self-fertilization. To attain this objective, laboratory-raised strains of B. glabrata and B. straminea were used. The former originated from natural breeding grounds in the municipality Paulista, state of Pernambuco, Brazil. The latter originated from irrigation ditches in the municipality of Petrolândia, in the same state. Snail populations of B. glabrata and B. straminea were maintained for 240 days in laboratory. Their fecundity was evaluated by noting the number of egg-masses, eggs and eggs per mass. Their fertility was evaluated by the number of viable eggs and the hatching rate. B. straminea was markedly more fecund than B. glabrata through cross- and self-fertilization, namely: greater egg-mass; higher egg production and more eggs per mass. Regarding fertility, there seemed to be no preferential period for occlusion to occur or a trend in the rhythm of producing viable eggs.