Embryonic development of teleost Brycon orbignyanus.

Brycon orbignyanus is an important large teleost that is currently on the list of endangered species, therefore studies on its reproductive biology and embryology are fundamental to help species conservation and recovery. The objective of this research was to characterize the events that occur during extrusion, fertilization and embryonic development of the species. The samples were collected at predetermined times, fixed and processed for light microscopy and scanning electron microscopy. The greenish oocytes were spherical, had translucent chorion and a mean diameter of 1.3±0.11 mm. The eggs had well defined animal and vegetative poles approximately 18 min post-fertilization. Stages from 2 to 128 blastomeres occurred between 20 min and 3 h post-fertilization (hPF), when the morula was characterized. The blastula stage was observed between 2 and 3 hPF, and the gastrula between 3 and 7 hPF, when the embryonic shield emerged and the cellular migration with the consequent formation of epiblast and hypoblast. At 8 hPF, the formation of the neural tube, above the notochord and the encephalic region, was observed, delimiting the forebrain, mesencephalon and rhombencephalon regions. From 11 hPF onward, the optic vesicle was formed close to the forebrain and the embryo tail was well developed. The optic vesicle was observed from 12 hPF onward, and the tail showed an intense movement that culminated with the rupture of the chorion and consequent hatching of the larva at 13 hPF and 27°C.

Vascular budding in Symplegma brakenhielmi and the evolution of coloniality in styelid ascidians.

Individuals of colonial animals (e.g. zooids) are in continuous turnover. In ascidians colonial or solitary species have evolved by convergence multiple times. Colonial Botryllus and Botrylloides are well-studied genera that exhibit colony-wide developmental mechanisms that regulate synchronous and orchestrated cycles of budding and turnover of zooids. The origins of modular developmental mechanisms that facilitated the evolution of coloniality in this group remain unclear. To reconstruct ancestral states of coloniality we studied Symplegma brakenhielmi, a sister taxon of the botryllids. S. brakenhielmi zooids are embedded in a common tunic and present a similar vascular system as the botrylloides, however development and turnover of zooids occurs asynchronously and in a more independent manner. We generated a table of common stages of budding in Symplegma and Botryllus for comparative studies of asexual development. We tested dependent processes of budding among individuals of the colony by systemic bud or zooid removals. Although our results showed a higher degree of independence in bud development in S. brakenhielmi, we found a subtle colony-wide regulatory mechanism of modular development, i.e. new buds expedited development after the removal of all buds in the colony. Next, we characterized external morphology, ultrastructure, and abundance of circulatory blood cells in the vascular system of S. brakenhielmi. Macrophage-like cells (MLCs) are involved in zooid resorption and turnover. Proportions of MLCs in the blood of S. brakenhielmi corresponded to the peak of occurrence of this cell type during the budding cycle of B. schlosseri. We found several new blood cell types in S. brakenhielmi, including two cell types that resemble circulatory progenitor stem cells of other botryllid colonial ascidians. These cells showed features of undifferentiated cells and expressed mitotic marker Phospho-histone H3. Comparative studies of S. brakenhielmi and B. schlosseri allow us to discuss possible changes in the regulation of modular development (i.e. regulation of life and death in the colony), and a possible contribution of circulatory blood cells in budding processes. We propose that the higher degree of developmental independence in S. brakenhielmi budding is a result of its ancestral solitary mode of development.

Structural and ultrastructural description of larval development in Zungaro jahu.

The Zungaro jahu is an important large catfish of the order Siluriformes that is in danger of extinction due to habitat destruction. Studies on its biology are scarce and the majority relates only to nutrition or parasitology. In order to provide greater information on its morphology and aid husbandry and larviculture studies, the aim of this study was to characterize larval development in Z. jahu from hatching to total yolk absorption. Samples were collected at pre-established times, processed, stained, and analyzed under stereomicroscopy, light microscopy, and scanning electron microscopy. Total yolk absorption was observed by 60 hours post-hatching (hph) at 28.75 ± 0.59°C. The newly hatched larvae showed slightly pigmented body, the outline of the digestive tract, evident eyes, and the first swimming movements. Mouth opening took place at 12 hph and the connection between the oral cavity and the rudimentary intestine was observed at 24 hph. Were analyzed the main larval organs and systems: digestive organs, heart, gill arches, sensory system, thyroid, kidney, and swim bladder. As the larvae grew, these organs became more mature and functional. The development of the sensory and feeding structures was observed at the start of larval development, and thus before depletion of endogenous energy reserves, the strategy for this species is to increase its chances of survival in the environment.

Embryonic development in Zungaro jahu.

The aim of this study was to characterize the embryonic development of Zungaro jahu, a fresh water teleostei commonly known as 'jaú'. Samples were collected at pre-determined times from oocyte release to larval hatching and analysed under light microscopy, transmission electron microscopy and scanning electron microscopy. At the first collection times, the oocytes and eggs were spherical and yellowish, with an evident micropyle. Embryo development took place at 29.4 ± 1.5°C and was divided into seven stages: zygote, cleavage, morula, blastula, gastrula, organogenesis, and hatching. The differentiation of the animal and vegetative poles occured during the zygote stage, at 10 min post-fertilization (mpf), leading to the development of the egg cell at 15 mpf. From 20 to 75 mpf, successive cleavages resulted in the formation of 2, 4, 8, 16, 32 and 64 blastomeres. The morula stage was observed between 90 and 105 mpf, and the blastula and gastrula stage at 120 and 180 mpf; respectively. The end of the gastrula stage was characterized by the presence of the yolk plug at 360 mpf. Organogenesis followed, with differentiation of the cephalic and caudal regions, elongation of the embryo by the cephalo-caudal axis, and somitogenesis. Hatching occurred at 780 mpf, with mean larval total length of 3.79 ± 0.11 mm.

Alterations in zebrafish development induced by simvastatin: Comprehensive morphological and physiological study, focusing on muscle.

The cholesterol synthesis inhibitor simvastatin, which is used to treat cardiovascular diseases, has severe collateral effects. We decided to comprehensively study the effects of simvastatin in zebrafish development and in myogenesis, because zebrafish has been used as a model to human diseases, due to its handling easiness, the optical clarity of its embryos, and the availability of physiological and structural methodologies. Furthermore, muscle is an important target of the drug. We used several simvastatin concentrations at different zebrafish developmental stages and studied survival rate, morphology, and physiology of the embryos. Our results show that high levels of simvastatin induce structural damage whereas low doses induce minor structural changes, impaired movements, and reduced heart beating. Morphological alterations include changes in embryo and somite size and septa shape. Physiological changes include movement reduction and slower heartbeat. These effects could be reversed by the addition of exogenous cholesterol. Moreover, we quantified the total cell number during zebrafish development and demonstrated a large reduction in cell number after statin treatment. Since we could classify the alterations induced by simvastatin in three distinct phenotypes, we speculate that simvastatin acts through more than one mechanism and could affect both cell replication and/or cell death and muscle function. Our data can contribute to the understanding of the molecular and cellular basis of the mechanisms of action of simvastatin.

The Unique Dorsal Brood Pouch of Thermosbaenacea (Crustacea, Malacostraca) and Description of an Advanced Developmental Stage of Tulumella unidens from the Yucatan Peninsula (Mexico), with a Discussion of Mouth Part Homologies to Other Malacostraca.

The Thermosbaenacea, a small taxon of crustaceans inhabiting subterranean waters, are unique among malacostracans as they brood their offspring dorsally under the carapace. This habit is of evolutionary interest but the last detailed report on thermosbaenacean development is more than 40 years old. Here we provide new observations on an ovigerous female of Tulumella unidens with advanced developmental stages in its brood chamber collected from an anchialine cave at the Yucatan Peninsula, which is only the third report on developmental stages of Thermosbaenacea and the first for the genus Tulumella. Significant in a wider crustacean context, we report and discuss hitherto unexplored lobate structures inside the brood chamber of the female originating at the first (maxilliped) and second thoracic segments, which are most likely modified epipods, perhaps serving as gills. At the posterior margin of carapace of the female are rows of large spines preventing the developing stages from falling out. The external morphology of the advanced developmental stages is described in much detail, providing information on e.g., carapace formation and early limb morphology. Among the hitherto unknown structures in the advanced developmental stages provided by this study are the presence of an embryonic dorsal organ and rudimentary 'naupliar processes' of the second antennae. Since most hypotheses on crustacean (and malacostracan and peracaridan) relationship rest on external limb morphology, we use early limb bud morphology of Tulumella to better establish thermosbaenacean limb homologies to those of other crustaceans, which is a necessary basis for future morphology based phylogenetic considerations.

Structural analysis of embryogenesis of Leiarius marmoratus (Siluriformes: Pimelodidae).

Embryological studies in fish species are useful to the understanding of their biology and systematics. The available biological data in Leiarius marmoratus are scarce and additional information about its reproductive biology is needed, mainly because this species has been commercially exploited and used in production of hybrid lineages. In order to evaluate the temporal-morphological embryonic modifications in L. marmoratus, samples of nearly 200 embryos were collected at random at different stages of development, starting from fecundation (time zero). Embryos were fixed in modified Karnovsk's solution and 2.5% glutaraldehyde, processed and analysed under optic and electron microscopy. The incubation period of L. marmoratus was equal to 14.42 h at a mean temperature of 28.3 ± 0.07°C. The following stages of embryonic development were established: zygote, cleavage, gastrula, organogenesis and hatching. These stages were divided into phases, as follows: cleavage - phases of 2, 4, 8, 16, 32 and 64 cells and morula; gastrula - phases of 25, 50, 75 and 90% of epiboly and blastopore closure; and organogenesis - neurula, segmentation and pre-larval phases. The embryogenesis of L. marmoratus was typical of neotropical teleosteans, with peculiarities in species development.

Injuries in pacu embryos (Piaractus mesopotamicus) after freezing and thawing.

Although the sperm cryopreservation of freshwater and marine teleosts has been feasible for years, the cryopreservation of some fish embryos still remains elusive. Thus, the objective of this experiment was to analyze the embryo morphology after freezing and thawing 40 embryos of Piaractus mesopotamicus immersed into methanol and ethylene glycol, both at 7, 10 and 13% plus 0.1 M sucrose for 10 min. Soon after thawing, three embryos were treated with historesin, stained with hematoxylin-eosin and analyzed under an optical microscope. From every treatment, one palette containing embryos was thawed and incubated, but none of the eggs hatched. Samples containing two embryos were immersed into 10% methanol or 10% ethylene glycol both in association with sucrose, and embryos immersed into only water or sucrose solution were frozen, processed and analyzed using scanning electron microscopy (SEM). In both cases, the control group was immersed into only water. Although the embryos had the chorion, vitello, yolk syncytial layer and blastoderm, all of them were found altered under the optical microscope and by SEM. The chorion was irregular and injured; there was no individuality in the yolk granules; the yolk syncytial layer had an irregular shape, thickness and size; the blastoderm showed injuries in the nucleus shape and sometimes was absent; the blastoderm was located in atypical areas and absent in some embryos. In conclusion, no treatment was effective in preserving the embryos, and none of the embryos avoided injury from intracellular ice formation. These morphological injuries during the freezing process made the P. mesopotamicus embryos unfeasible for hatching.

Increasing storage capability of pacu (Piaractus mesopotamicus) embryos by chilling: development of a useful methodology for hatcheries management.

Cryopreservation of fish gametes has been studied extensively in the last few decades, but the successful cryopreservation of fish embryos remains elusive. However, recent studies using short-term chilling techniques have shown that it is possible to store embryos at low temperatures with no significant loss in viability. Information on cryopreservation of Neotropical freshwater fish embryos has so far been very limited in the literature. In the present study, chilling protocols for storage of pacu embryos at -8°C for up to 24 h were studied using different concentrations of sucrose in methanol. Embryos tolerated the subzero temperature for up to 6 h with no adverse effects (P > 0.05). After 12 h chilling, hatching rate of 64.0 +/- 3.5 percent was recorded. Low temperature storage of pacu embryos by chilling is detailed here for the first time. Further studies are needed to extend the storage time and to improve the hatching rate.

Muscle development and differentiation in the urodele Ambystoma mexicanum.

Muscle differentiation has been widely described in zebrafish and Xenopus, but nothing is known about this process in amphibian urodeles. Both anatomical features and locomotor activity in urodeles are known to show intermediate features between fish and anurans. Therefore, a better understanding of myogenesis in urodeles could be useful to clarify the evolutionary changes that led to the formation of skeletal muscle in the trunk of land vertebrates. We report here a detailed morphological and molecular investigation on several embryonic stages of Ambystoma mexicanum and show that the first differentiating muscle fibers are the slow ones, originating from a myoblast population initially localized close to the notochord that forms a superficial layer on the somitic surface afterwards. Subsequently, fast fibers differentiation ensues. We also identified and cloned A. mexicanum Myf5 as a muscle-specific transcriptional factor likely involved in urodele muscle differentiation.

Morphometric characterization of the first blastomeres of rainbow trout (Oncorhynchus mykiss).

In the following investigation the morphometric characteristics of the first two blastomeres of rainbow trout (Oncorhynchus mykiss) were determined. Embryos were incubated at 9°C and then fixed in a Stockard solution every 30 min starting from 8.5 to 12.5 h of incubation post fertilization. Embryonic discs were extracted and microphotographs were taken with Q Capture Pro 5.0 software using a stereomicroscope Olympus SZX7. The average size of the blastodiscs was 941.22 ± 160.42 µm. The first cleavage finished after approximately 12 h of incubation. The first two blastomeres were regularly symmetrical in their morphology. Blastomere 1 had an average length (L) of 942.68 ± 105.56 µm and width (W) of 467.34 ± 64.33 µm. Blastomere 2 had an average length of 887.60 ± 101.65 and width of 454.49 ± 47.25 µm (n = 91). Significant differences were found between the length and width of blastomeres 1 and 2. The proportion between the length of blastomeres 1 and 2 was 0.94 ± 0.07 (n = 91); between the width of blastomeres 1 and 2 it was 0.88 ± 0.11 (n = 91); and the width/length ratio was 0.51 ± 0.09 (n = 182). It was concluded that rainbow trout blastomeres tend to be asymmetrical in length with a higher dispersion of widths.

Eggs ultrastructure and early development of Franciscodoras marmoratus (Pisces: Doradidae).

This study presents, for the first time, information on the eggs and early development of Franciscodoras marmoratus, fish of São Francisco river, Brazil. To analyse the egg ultrastructure and morphological events of embryogenesis, a total of 36 F. marmoratus specimens (18 males and 18 females) were captured and subjected to spawning induction. Gametes were collected by manual extrusion, and fertilization was conducted using the dry method. After fertilization, eggs were kept in incubators with water temperature of 24°C. The embryonic development was monitored using a stereomicroscope until hatching. There was a 67% positive response to hypophysation by the females and the fertilization rate was 73.8 ± 6.2%. The oocytes are discoid, yellow, adhesive and covered by a thick jelly coat. Under the electron scanning microscope, the oocytes presented a surface with pore canals and funnel-shaped micropyle with a smooth vestibule. Recently extruded oocytes had a mean diameter of 1.27 ± 0.4 mm and after hydration was 1.91 ± 0.05 mm. The jelly coat was 0.34 ± 0.03 mm thickness, and the perivitelline space was 0.19 ± 0.04 mm. Eight phases of the embryonic development were identified, and embryogenesis was completed at 47 h after fertilization, at 24°C water temperature. The recently hatched larvae had 2.76 ± 0.57 mm of total length. These results provide useful information for the successful breeding and reproductive strategies of fishes.

Early development of Astronotus ocellatus under stereomicroscopy and scanning electron microscopy.

Astronotus ocellatus, popularly known as Oscar, is a cichlid fish from the Amazon basin (Brazil) with a great potential for fish farming. The aim of this research is to describe the morphology of eggs and larvae of A. ocellatus under stereomicroscopy and scanning electron microscopy. Eggs from natural spawnings were taken to hatcheries, collected at previously established time periods and then analysed. Oscar's eggs are demersal, adhesive and fragile to touch, with a slightly oval shape. The fertile eggs are yellowish in colour and when unfertilized are a white opaque colour. In the initial collection (IC), the majority of eggs were found to be at the gastrula phase with 30% epiboly. At 12 h after the IC, the formation of the embrionary axis and somites was observed, followed by differentiation of the tail and of the head. Fifteen hours after the IC, the emergence of the optic and otic vesicles, and of adhesive glands and the yolk pigmentation was observed. Larval hatching took place between 46 and 58 h after the first collection, at an average temperature of 27.45 ± 2.13°C. The larval stage was characterized by the development of the heart, fins, branchial apparatus, neuromasts, taste buds and adhesive glands on the head. Larval development to yolk absorption took a period of 257 h. These results provide important information for reproduction, rearing and preservation of A. ocellatus.

Embryonic development of Anodontites trapesialis (Lamarck, 1819) (Bivalvia: Mycetopodidae).

The phases of embryonic development of Anodontites trapesialis lasidia are described for the first time. Adult specimens were obtained from two fish farms located in Londrina, Paraná, Brazil. The internal demibranchs of 120 individuals were studied using a routine histological technique; 70 of these carried eggs and/or larvae in the marsupium and were utilized for the description of the phases of embryonic development. The demibranchs of five specimens were evaluated by scanning electron microscopy to detail the morphology of the larvae. Five phases of development were established: phase I, corresponding to the initial stage of cleavage with the formation of apical cells; phase II, including the stages of the morula and blastula; phase III, where the gastrula forms; phase IV, where the larva formed is still inside the egg envelope; and phase V, where the lasidium can still be identified immediately after eclosion.

Ultrastructural development of the small intestinal mucosa in the embryo and turkey poult: A light and electron microscopy study.

The potential for growth and feed efficiency in turkey poults directly correlates with the early development of the intestinal epithelium. Although the metabolic aspects of enteric maturation have been studied, little is known about the ultrastructural development of the enteric epithelium in the turkey embryo and poult. Hence, the objective of this study was to document the morphological and ultrastructural development of the jejunum mucosa in turkeys, from 15 d of incubation (embryonic day; E) to 12 d posthatch. Intestinal samples from 4 embryos or poults were collected and analyzed by light and electron microscopy (transmission and scanning). In addition, amniotic fluid volume was determined in 6 eggs from E15 to E25. Longitudinal previllus ridges at E15 gradually formed zigzag patterns that led to the formation of 2 parallel lines of mature villi by E25. The volume of amniotic fluid was rapidly depleted as the embryo swallowed it between E19 and E25. During this period, a major increase occurs in villus height, the apical end of epithelial cells is gradually tightened by the junctional complex, and mature goblet cells are visible at the apical end of villi. Villus height steadily increases until reaching a plateau at 8 d. Villi morphology shifts gradually from finger-like projections before hatch to leaf-like projections by 12 d. At this age, the enteric epithelium is in intimate association with microbes such as segmented filamentous bacteria. The profound morphological adaptations of the turkey gut epithelium in response to amniotic fluid swallowing before hatch, and dietary factors and bacteria after hatch, demonstrate the plasticity of the enteric epithelium at this time. Hence, the supplementation of enteric modulators before hatch (in ovo feeding) and after hatch has the potential to shape gut maturation and enhance the growth performance of turkey poults.

Embryonic development of Anodontites trapesialis (Lamarck, 1819) (Bivalvia: Mycetopodidae) / Desenvolvimento embrionário de Anodontites trapesialis (Lamarck, 1819) (Bivalvia: Mycetopodidae)

The phases of embryonic development of Anodontites trapesialis lasidia are described for the first time. Adult specimens were obtained from two fish farms located in Londrina, Paraná, Brazil. The internal demibranchs of 120 individuals were studied using a routine histological technique; 70 of these carried eggs and/or larvae in the marsupium and were utilized for the description of the phases of embryonic development. The demibranchs of five specimens were evaluated by scanning electron microscopy to detail the morphology of the larvae. Five phases of development were established: phase I, corresponding to the initial stage of cleavage with the formation of apical cells; phase II, including the stages of the morula and blastula; phase III, where the gastrula forms; phase IV, where the larva formed is still inside the egg envelope; and phase V, where the lasidium can still be identified immediately after eclosion.

As fases do desenvolvimento embrionário das lasídias de Anodontites trapesialis são descritas pela primeira vez. Espécimes adultos foram obtidos de duas pisciculturas localizadas no município de Londrina, Paraná, Brasil. As demibrânquias internas de 120 indivíduos foram estudadas por técnicas histológicas rotineiras; 70 apresentavam ovos e/ou larvas no marsúpio e foram utilizadas para a descrição das fases do desenvolvimento embrionário. As demibrânquias de cinco espécimes foram avaliadas por microscopia eletrônica de varredura para detalhar a morfologia da larva. Cinco fases do desenvolvimento foram estabelecidas: fase I, correspondente ao estágio inicial da clivagem com formação das células apicais; fase II, incluindo os estágios de mórula e blástula; fase III, na qual se forma a gástrula; fase IV, na qual a larva formada ainda encontra-se dentro do ovo; e fase V, na qual a lasidium pode ser identificada imediatamente após a eclosão.

Developmental biology of the Brazilian 'Armed' spider Phoneutria nigriventer (Keyserling, 1891): microanatomical and molecular analysis of the embryonic stages.

Phoneutria (Ctenidae) is among the most dangerous venomous spiders in Brazil. Its venom is composed of a mixture of pharmacologically active components, some of which have been quite extensively studied due to their potentiality as models for new pharmaceutical drugs. Nevertheless, literature data on the venom-producing glands are very limited. In the present study, we follow the biological development of intra-cocoon stages of Phoneutria nigriventer spiders, mainly regarding the formation of the venomous apparatus and venom production. The results showed that the venom glands of Phoneutria are already present in the early 1st pre-larva stage. The venomous apparatus is completely formed in the larva, a stage that precedes the spider eclosion from the cocoon. At embryo stages, transcripts of a vertebrate-active neurotoxin (PhTx1) were shown to be present, as well as, unidentified venom proteins that were immunolabeled by anti-venom antibodies. It seems that venom toxins play roles in the protection and survival of those early developmental stages of Phoneutria spiders.

Freezing injuries in the embryos of Piaractus mesopotamicus.

Cryopreservation of mammal embryos has been technically feasible for many years, but morphological injuries still persist in fish embryos during cryopreservation. Thus, the objective of the present study was to describe these freezing injuries in Piaractus mesopotamicus embryos. Two hundred and twenty-five embryos were collected at the post-gastrula stage and assigned into four treatments of sucrose at 8.5, 17.0, 25.0 or 34.0% plus 9.0% methanol. The control was prepared with distilled water only. The gradual decrease in the temperature was 0.5°C/min. After the seeding stage, the fish embryos were stored in liquid nitrogen at -33°C. Thereafter, they were thawed for evaluating per cent hatching, and the samples collected from every treatment were submitted to scanning electron microscopy for morphological analysis. The micrographic images showed that there was substantial alterations in embryo morphology under the highest concentrations of sucrose. These solutions did not prevent the formation of ice crystals, which lead to deformities and killed the embryos, but the observed reduced level of morphological structure in these embryos when treated with 17.0% sucrose plus 9.0% methanol is a compelling argument for additional studies.

Electron microscopy of the amphibian model systems Xenopus laevis and Ambystoma mexicanum.

In this chapter we provide a set of different protocols for the ultrastructural analysis of amphibian (Xenopus, axolotl) tissues, mostly of embryonic origin. For Xenopus these methods include: (1) embedding gastrulae and tailbud embryos into Spurr's resin for TEM, (2) post-embedding labeling of methacrylate (K4M) and cryosections through adult and embryonic epithelia for correlative LM and TEM, and (3) pre-embedding labeling of embryonic tissues with silver-enhanced nanogold. For the axolotl (Ambystoma mexicanum) we present the following methods: (1) SEM of migrating neural crest (NC) cells; (2) SEM and TEM of extracellular matrix (ECM) material; (3) Cryo-SEM of extracellular matrix (ECM) material after cryoimmobilization; and (4) TEM analysis of hyaluronan using high-pressure freezing and HABP labeling. These methods provide exemplary approaches for a variety of questions in the field of amphibian development and regeneration, and focus on cell biological issues that can only be answered with fine structural imaging methods, such as electron microscopy.

Embryonic development of Ampheres leucopheus and Iporangaia pustulosa (Arachnida: Opiliones: Gonyleptidae).

The first studies concerning the embryonic development of harvestmen started in the late 19th century, and focused mostly on holarctic species, and only three species of the suborder Laniatores (the largest, among the four suborders considered presently) were studied. Moreover, the last studies on embryology of harvestmen were made during the late 1970s. This study focused on the embryonic development of Ampheres leucopheus (Gonyleptidae, Caelopyginae) and Iporangaia pustulosa (Gonyleptidae, Progonyleptoidellinae). The embryonic development was followed in the field, by taking daily photographs of different eggs during about 2 months. When laid, eggs of A. leucopheus and I. pustulosa have approximately 1.13 and 1.30 mm in diameter, respectively, and the second is embedded in a large amount of mucus. The eggs grow, mainly due to water absorption at the beginning of the process, and they reach a diameter of about 1.35 and 1.59 mm, respectively, close to hatching. It took, respectively, 29-56 days and 35-66 days from egg laying to hatching. For the description of the embryonic development, we use photographs from the field, SEM micrographs, and histological analysis. This allowed us, for instance, to document the progression of structures and pigmentation directly from live embryos in the field, and to record microstructures, such as the presence of perforations in the cuticle of the embryo in the place where eyes are developing. Yet, contrary to what was expected in the literature, we record an egg tooth in one of the studied laniatoreans.