Days Of Larval Development Research Articles

Balancing selection and candidate loci for survival and growth during larval development in the Mediterranean mussel, Mytilus galloprovincialis.

Many marine bivalves have complex life histories with distinct developmental processes and genetic mechanisms. Larval development for most bivalves is often a prolonged and crucial physiological stage, where they suffer mass mortality due to early-acting genetic load. In this study, we describe genetic changes taking place within a single generation of families of the Mediterranean mussel Mytilus galloprovincialis over 23 days of larval development. Using replicated cultures and a pooled sequencing approach, we demonstrate that temporal balancing selection at the majority of loci preserve genetic variation in the early developmental stages of M. galloprovincialis. Balancing selection may be the mechanism which maintains standing genetic variation within the mussel genome and may improve the chances of survival and shield larvae from high levels of genetic load. Additionally, we used changes in allele frequencies to identify potential size-associated SNPs and viability-associated SNPs and found that patterns of genetic changes in directionally selected SNPs cannot be simply explained by traditional theories of genetic purging or directional selection without consideration of balancing selection. Finally, we observed a negative correlation between larval growth rates and survival, implying a potential trade-off relationship between the 2 commercially relevant phenotypes.

Embryonic and Larval Development of Thai Pangas Pangasianodon Hypopthalmus (Sauvage, 1878) Under Hatchery Condition

The embryonic and larval development of Pangasianodon hypophthalmus was investigated during peak spawning periods (May-July 2019). The fertilized eggs were adhesive and spherical with a greenish‐brown egg capsule. The yolk sac was yellowish‐brown in color and 1.00–1.70 mm in diameter. The first cleavage stage, embryonic shield, head, tail region, neural grooves and somites were apparent after fourteen hours post‐fertilization. The ranges of incubation period were from 18.00-20.00 hrs at a temperature of 26-30°C. The newly hatched larvae were transparent and light yellowish in color with a body length of 2.98–3.10 mm. Eye pigment appeared and the heart started to work within 12-16 hrs of hatching. The mouth became well developed at the age of 24 hrs; barbules were prominent, elongated and look like threads. The yolk sac was fully absorbed and the palatine teeth were fully increased during the 3 days pro‐larval stage. The stomach became functional and aerial respiration started after 3 days of larval development. After 72 hrs, the young fry was well‐developed, and appeared an adult, and length was measured up to 7.20±0.02 mm. When daily ration of pangas larvae is insufficient cannibalism was recorded. This study must be supportive for researcher and nursery owner on the biology and ecology of the fish, which might be helpful for appropriate measure of sustainable nursery, rearing and management technology of pangas. Bangladesh J. Zool. 49(1): 147-159, 2021

B-vitamin nutrition in the pea aphid-Buchnera symbiosis

Various insects that utilize vitamin-deficient diets derive a supplementary supply of these micronutrients from their symbiotic microorganisms. Here, we tested the inference from genome annotation that the symbiotic bacterium Buchnera aphidicola in the pea aphid Acyrthosiphon pisum provides the insect with vitamins B2 and B5 but no other B-vitamins. Contrary to expectation, aphid survival over five days of larval development on artificial diets individually lacking each B-vitamin not synthesized by Buchnera was not significantly reduced, despite significantly lower carcass B1, B3, B6 and B7 concentrations in the aphids on diets lacking each of these B-vitamins than on the vitamin-complete diet. Aphid survival was, however, significantly reduced on diet containing low concentrations (≤0.2 mM) or no pantothenate (B5). Complementary transcriptome analysis revealed low abundance of the sense-transcript, but high abundance of the antisense transcript, of the Buchnera gene panC encoding the enzyme mediating the terminal reaction in pantothenate synthesis. We hypothesize that metabolic constraints or antisense transcripts may reduce Buchnera-mediated production of pantothenate, resulting in poor aphid performance on pantothenate-free diets. The discrepancy between predictions from genome data and empirical data illustrates the need for physiological study to test functional inferences made from genome annotations.

Evidence of Oxidative Phosphorylation in Zebrafish Photoreceptor Outer Segments at Different Larval Stages.

Previous studies on purified bovine rod outer segments (OS) disks pointed to Oxidative Phosphorylation (OXPHOS) as being the most likely mechanism involved in ATP production, as yet not fully understood, to support the first phototransduction steps. Bovine and murine rod OS disks, devoid of mitochondria, would house respiratory chain complexes I to IV and ATP synthase, similar to mitochondria. Zebrafish ( Danio rerio) is a well-suited animal model to study vertebrate embryogenesis as well as the retina, morphologically and functionally similar to its human counterpart. The present article reports fluorescence and Transmission Electron Microscopy colocalization analyses of respiratory complexes I and IV and ATP synthase with zpr3, the rod OS marker, in adult and larval zebrafish retinas. MitoTracker Deep Red 633 staining and assays of complexes I and III-IV activity suggest that those proteins are active in OS. Results show that an extramitochondrial aerobic metabolism is active in the zebrafish OS at 4 and 10 days of larval development, as well as in adults, suggesting that it is probably maintained during embryogenesis. Data support the hypothesis of an extramitochondrial aerobic metabolism in the OS of zebrafish.

Microbial composition affects the performance of an artificial Tephritid larval diet.

The present study investigated the patterns of microorganisms in an artificial larval diet during Dacus ciliatus (Diptera; Tephritidae) larval development. Microbial population contents in the diet of total heterotrophic bacteria, yeast and molds, coliform and lactobacilli, and their dynamics during development, were monitored. Initially, the microbial composition in diet trays failing to produce viable pupae and in trays successfully producing pupae and adult flies was characterized. The failing diet trays contained large populations of lactobacilli that increased during larval development, and low populations of coliforms. In contrast, the successful diet showed an increasing population of coliforms and a low, or undetected, population of lactobacilli. To study the hypothesis that lactobacilli affect D. ciliatus larval development, we conducted controlled inoculation experiments in which Lactobacillus plantarum was added into fresh diet at the time of egg seeding. L. plantarum inoculated trays showed no production of D. ciliatus. Control trays without lactobacilli inoculation showed variable results. One tray successfully produced viable pupae and adults, and showed a slight and slow increase in the indigenous populations of lactobacilli. The second tray, however, failed to produce pupae and showed a fast increase of the indigenous lactobacilli to very high levels. Monitored pH trends in L. plantarum-inoculated diet showed a sharp pH decrease during the first 4 days of larval development from 5 to less than 4 units, while successful diet, producing viable D. ciliatus pupae and adults, showed a moderate pH drop during most of the larval development period. The paper discusses the possible ecological interactions between D. ciliatus larvae, the microbial content of the diet and the physical properties of the diet. The discussion also points out at the usefulness of this approach in understanding and managing mass production parameters of tephritid fruit flies industrial diets used for Sterile Insect Technique.

To divide or not to divide: An alternative behavior for teratocytes in Encarsia pergandiella (Hymenoptera: Aphelinidae)

Encarsia pergandiella (Hymenoptera: Aphelinidae) is an endoparasitoid with an unusual embryonic development compared to most of congeneric species and all other members of the superfamily Chalcidoidea. The developmental background of this wasp is based on an alecithal hydropic egg, with the embryo developing inside an extra-embryonic membrane which dissociates at hatching into special larva-assisting cells, the teratocytes. In E. pergandiella many teratocytes at hatching were multinucleated syncytial cells with no evidence of a cellular membrane separating the nuclei. These teratocytes during larval development produced smaller uninucleated teratocytes, through successive divisions obtained by progressive ingrowth of the plasmatic membrane, accompanied by appearance of degeneration symptoms, such as protrusions and blebs. As a consequence of this divisional process teratocytes showed a size reduction and an increase in number of about four times during the second day of larval development. Only on the third day of larval life teratocytes started to decrease in number, until total disappearance at larval maturation. This behaviour is in striking contrast with all other studied systems in which teratocytes do not divide and progressively decrease in number as the parasitoid larva develops.

Natural Larval Diet Differently Influences the Pattern of Developmental Changes in DNA 5-Methylcytosine Levels in Apis mellifera Queens as Compared with Workers and Drones.

The principal mechanism of gene activation/silencing is DNA 5-methylcytosine methylation. This study was aimed at determining global DNA methylation levels in larvae, prepupae, pupae, and 1-day-old adults of Apis mellifera queens, workers and drones. The Imprint Methylated DNA Quantification Kit MDQ1 was used. Percentages of DNA 5-methylcytosine were low and relatively similar in the larvae of all the castes until 4th day of larval development (3-5%). However, they were higher in the drone and worker larvae than in the queen larvae. Generally, the developmental patterns of changes in the DNA methylation levels were different in the queens in comparison with the drones and workers. While methylation increased in the queens, it decreased in the drones and workers. Methylated DNA methylcytosine percentages and weights in the queen prepupae (15%, 9.18ng) and pupae (21%, 10.74ng) were, respectively, three and four times higher than in the worker/drone brood of the same age (2.5-4%, 0.03-0.07ng). Only in the queens, after a substantial increase, did DNA methylation decrease almost twice between the pupal stage and queen emergence (from 21% and 10.74ng to 12% and 6.78ng). This finding seems very interesting, particularly for experimental gerontology.

Activin signaling balances proliferation and differentiation of ovarian niche precursors and enables adjustment of niche numbers.

How the numbers of niches and resident stem cells within a particular organ are determined during development and how they may be modulated or corrected is a question with significant medical implications. In the larval ovary of Drosophila melanogaster, somatic precursors for niches, and germ cells that will become germline stem cells, co-develop. Somatic precursors proliferate during the first 3 days of larval development. By mid-third instar, adult terminal filament (TF) (part of the germline stem cell niche) cells first appear, and differentiation terminates 24 h later when 16-20 TFs fully form. The developmental sequence responsible for TF cell determination and final TF numbers is only partially understood. We show that TF formation proceeds through several, hitherto uncharacterized stages, which include an early exit from the cell cycle to form TF precursors and two steps of cell shape change to form the mature TF cells. The Activin receptor Baboon (Babo) is required for somatic precursor cell proliferation and therefore determines the pool of TF precursors available for TF differentiation. During the final differentiation stage, Babo facilitates TF and germ cell differentiation, and promotes the accumulation of Broad-Z1, which is also a target of the steroid hormone ecdysone. Epistasis analysis shows that Activin controls cell proliferation in an ecdysone-independent manner and TF differentiation by affecting ecdysone targets. We propose that this mode of function allows Activin to balance proliferation and differentiation, and to equilibrate niche numbers. These results suggest a novel model for how niche numbers are corrected during development.

Recipe for a Busy Bee: MicroRNAs in Honey Bee Caste Determination

Social caste determination in the honey bee is assumed to be determined by the dietary status of the young larvae and translated into physiological and epigenetic changes through nutrient-sensing pathways. We have employed Illumina/Solexa sequencing to examine the small RNA content in the bee larval food, and show that worker jelly is enriched in miRNA complexity and abundance relative to royal jelly. The miRNA levels in worker jelly were 7–215 fold higher than in royal jelly, and both jellies showed dynamic changes in miRNA content during the 4th to 6th day of larval development. Adding specific miRNAs to royal jelly elicited significant changes in queen larval mRNA expression and morphological characters of the emerging adult queen bee. We propose that miRNAs in the nurse bee secretions constitute an additional element in the regulatory control of caste determination in the honey bee.

Short- and long-term consequences of larval stage exposure to constantly and ephemerally elevated carbon dioxide for marine bivalve populations

Abstract. While larval bivalves are highly sensitive to ocean acidification, the basis for this sensitivity and the longer-term implications of this sensitivity are unclear. Experiments were performed to assess the short-term (days) and long-term (months) consequences of larval stage exposure to varying CO2 concentrations for calcifying bivalves. Higher CO2 concentrations depressed both calcification rates assessed using 45Ca uptake and RNA : DNA ratios in Mercenaria mercenaria and Argopecten irradians larvae with RNA : DNA ratios being highly correlated with larval growth rates (r2>0.9). These findings suggested that high CO2 has a cascading negative physiological impact on bivalve larvae stemming in part from lower calcification rates. Exposure to elevated CO2 during the first four days of larval development significantly depressed A. irradians larval survival rates, while a 10-day exposure later in larval development did not, demonstrating the extreme CO2 sensitivity of bivalve larvae during first days of development. Short- (weeks) and long-term (10 month) experiments revealed that individuals surviving exposure to high CO2 during larval development grew faster when exposed to normal CO2 as juveniles compared to individuals reared under ambient CO2 as larvae. These increased growth rates could not, however, overcome size differences established during larval development, as size deficits of individuals exposed to even moderate levels of CO2 as larvae were evident even after 10 months of growth under normal CO2 concentrations. This "legacy effect" emphasizes the central role larval stage CO2 exposure can play in shaping the success of modern-day bivalve populations.

An advance in the in vitro rearing of stingless bee queens

Stingless bees are a diverse group of highly eusocial tropical bees and potential pollinators of natural and crop environments. Nevertheless, large-scale breeding for agricultural purposes still needs development. A bottleneck is the small number of queens generally found in the colonies of most species. A proposed solution is to develop in vitro rearing of stingless bee queens, which includes a technical support due to the massive liquid feeding of larvae. In this study, we tested the efficacy of in vitro queen-rearing techniques in terms of queen size, survival, and fecundity. We found that humidity must be kept high during the first 6 days of larval development and reduced thereafter to 75 %. In the most effective treatments, we obtained up to 97.9 % queen survival. We also found that queens not different in size from natural queens could be produced if sufficient larval food is provided. In vitro queens were able to mate and lay viable eggs. This methodology is also useful for several research appliances, like pesticide effect on bees, pollen quality effect in caste determination, and the use of genetic sources for selection programs, among others.

Correlated basal expression of immediate early gene egr1 and tyrosine hydroxylase in zebrafish brain and downregulation in olfactory bulb after transitory olfactory deprivation

Imprinting on kin occurs during the sixth day of larval development in zebrafish and depends on olfactory signals. In rodents, the immediate early gene egr1 is involved in maintaining the dopaminergic phenotype of periglomerular olfactory bulb cells in an activity dependent way. Furthermore, egr1 is upregulated in medial amygdalar dopamine cells in some rodents (prairie voles) dependent on social pheromone interactions. Thus, we aimed to investigate whether egr1 is involved in imprinting processes and later kin recognition in zebrafish in olfactory centers, such as the olfactory bulb and suspected medial amygdala. In the present paper, we focus on a basic investigation of basal egr1 expression throughout zebrafish brain development and its co-localization with tyrosine hydroxylase as a marker for dopaminergic neurons. Indeed, there is unambiguous co-localization of egr1 and tyrosine hydroxylase in the zebrafish olfactory bulb and hypothetical medial amygdala. Furthermore, as in rodents, ipsilateral transient olfactory deprivation through Triton X-100 treatment of the olfactory epithelium leads to downregulation of egr1 and tyrosine hydroxylase expression in the olfactory bulb, but apparently not in secondary olfactory targets of the zebrafish brain. This indicates that similar processes might be at work in zebrafish and rodent olfactory systems, but their more specific involvement in imprinting in zebrafish has to be further tested.

Embryonic development and larval stages of Steindachneridion parahybae (Siluriformes: Pimelodidae): implications for the conservation and rearing of this endangered Neotropical species

Steindachneridion parahybae is a freshwater catfish endemic to the Paraíba do Sul River and is classified as an endangered Neotropical species. An increasing number of conservation biologists are incorporating morphological and physiological research data to help conservation managers in rescue these endangered species. This study investigated the embryonic and larval development of S. parahybae in captivity, with emphasis in major events during the ontogeny of S. parahybae. Broodstocks were artificially induced to reproduce, and the extrusion occurred 200-255 degree-hours after hormonal induction at 24°C. Larval ontogeny was evaluated every 10 minutes under microscopic/stereomicroscopic using fresh eggs samples. The main embryogenic development stages were identified: zygote, cleavage, including the morula, blastula, gastrula phase, organogenesis, and hatching. The extruded oocytes showed an average diameter of 1.10 ± 0.10 mm, and after fertilization and hydration of eggs, the average diameter of eggs increased to about 1.90 ± 0.60 mm, characterized by a large perivitelline space that persisted up to embryo development, the double chorion, and the poles (animal and vegetative). Cell division started about 2 minutes after fertilization (AF), resulting in 2, 4, 8 (4 x 2 arrangement of cells), 16 (4 x 4), 32 (4 x 8) and 64 (2 x 4 x 8) cells. Furthermore, the blastula and gastrula stages followed after these cells divisions. The closed blastopore occurred at 11 h 20 min AF; following the development, the organogenetic stages were identified and subdivided respectively in: early segmentation phase and late segmentation phase. In the early segmentation phase, there was the establishment of the embryonic axis, and it was possible to distinguish between the cephalic and caudal regions; somites, and the optic vesicles developed about 20 h AF. Total hatching occurred at 54 h AF, and the larvae average length was 4.30 ± 0.70 mm. Gradual yolk sac reduction was observed during the first two days of larval development. The first feeding occurred at the end of the second day. During the larval phase, cannibalism, heterogeneous larval growth and photophobia were also observed. This information will be important in improving the artificial reproduction protocols of S. parahybae in controlled breeding programs.

Laboratory rearing and life history of an emerging grape pest, Xylotrechus arvicola (Coleoptera: Cerambycidae)

Several aspects of the biology of Xylotrechus arvicola (Olivier), an emerging grape pest, were studied under laboratory conditions. Four diets were tested to rear this species in the laboratory. Among them, only one made rearing from larva to adult possible. The highest mortality, in all cases, was recorded during the first days of larval development. Larvae were kept 45 days at 8°C to break diapause in order to reduce the normal field larval developmental time. The species' developmental time was similar between sexes, while pupal developmental time and weight were significantly greater for females than for males. As part of a complementary study, life table parameters of females obtained from the larvae reared on the artificial diet were compared to those of females emerged from field-infested grape root wood. Both laid the majority of eggs in the first two weeks after emergence, and they had a similar pre-laying period. Nevertheless, the females from the diet-reared larvae lived significantly longer, laid eggs over a longer period of time and showed higher fertility than those emerging from infested grape root, suggesting that diet fulfils larval nutritional needs. The species' laboratory-reared population exhibited a low intrinsic growth rate value (rm=0.01) as a result of its long egg-to-adult developmental time and its high larval mortality.

The tadpole of Ecnomiohyla sukia Savage & Kubicki, 2010 (Amphibia: Hylidae)

Near the village of Guayacán de Siquirres, Limón province, Costa Rica (N 10°02”, W 83°31’) the call of a frog revealed a breeding hole about 15 m high in a Guarea tree. An adult was taken from this water-filled cavity along with some tadpoles. The adult tree frog was identified by B. Kubicki as Ecnomiohyla sukia Savage and Kubicki, 2010. Its sex was not determined. Ten tadpoles were extracted from the water-filled tree hole and it appeared they had only a few days of larval development, but the exact age was undeterminable. The conspecificity of the tadpoles found with the adult frog E. sukia was established by rearing them to metamorphic stages and froglets. The following description is based on tadpoles from stages 25, 36–37, and 41 (Gosner 1960).

Molecular structure of the prothoracicotropic hormone gene in the northern house mosquito, Culex pipiens, and its expression analysis in association with diapause and blood feeding

We cloned the gene that encodes prothoracicotropic hormone (PTTH) in the northern house mosquito, Culex pipiens, and investigated its expression profile in short-day (diapause-destined) and long-day (nondiapause-destined) individuals from the fourth-instar larval stage to 2 months of adulthood, as well as after a blood meal. The deduced C. pipiens PTTH (Cupip-PTTH) amino acid sequence contains seven cysteines with a specific spacing pattern. Sequence alignment suggests that Cupip-PTTH is 23% identical to Drosophila melanogaster PTTH, but is ≥59% identical to the PTTHs of other mosquitoes. Cupip-PTTH has structural characteristics similar to those of Bombyx mori PTTH and some vertebrate nerve growth factors with cysteine-knot motifs. PTTH transcripts exhibit a daily cycling profile during the final (fourth) larval instar, with peak abundance occurring late in the scotophase. The fourth-larval instar stage is one day longer in short-day larvae than in long-day larvae, resulting in larger larvae and adults. This additional day of larval development is associated with one extra PTTH cycle. No cycling was observed in pupae, but PTTH transcripts were slightly higher in short-day pupae than in long-day pupae throughout much of the pupal stage. PTTH expression persisted at a nearly constant level in diapausing adult females for the first month but then dropped by ∼50%, while expression decreased at the beginning of adulthood in nondiapausing females and then remained at a low level as long as the females were denied a blood meal. However, when nondiapausing females were offered a blood meal, PTTH transcripts rose approximately 7 fold in 2 h and remained elevated for 24 h. A few diapausing females (∼10%) will take a blood meal when placed in close proximity to a host, but much of the blood is ejected and such meals do not result in mature eggs. Yet, elevated PTTH mRNA expression was also observed in diapausing females that were force fed. Our results thus point to several distinctions in PTTH expression between short-day and long-day mosquitoes, but both types of females responded to a blood meal by elevating levels of PTTH mRNA.

Development of digestive system and swim bladder of larval nase (Chondrostoma nasus L.)

Digestive tract and swim bladder differentiation during nase (Chondrostoma nasus L.) larval development was evaluated using histological methods. The mouth, pharynx and oesophagus were impervious at the time of hatching. The straight tubular intestine consisted of undifferentiated cells and was situated over the yolk sac. The intestine, lined with a single layer of columnar epithelial cells, opened on 2 day posthatching (dph). First evidence of protein digestion and absorption by the enterocytes was observed on the sixth day, while lipid digestion and absorption began on the ninth day of larval development. Intestinal mucous cells developed and became active by the fourth day. The posterior chamber of the swim bladder inflated on the third day, while the anterior one filled 12 dph. Complete yolk sac resorption took place on the ninth day.

Digestive enzymes during development of Ceratitis capitata (Diptera:Tephritidae) and effects of SBTI on its digestive serine proteinase targets

The digestive system of Ceratitis capitata was characterized during its larval development and in the insect stage. Disaccharidases against maltose and sucrose were more evident in the 2nd and 3rd day of larval development and in the adult stage, respectively. Glycosil-hydrolyses with higher specific α-galactosidasic and β-galactosidasic activities were detected in the 2nd and 3rd day of the larval stage, respectively. Specific proteolytic activities against azocasein showed an increase in the 4th and 5th day of the larval stage and in the adult stage. Specific hemoglobin activities were constant between 2nd and 6th day of the larval stage. The larvae used mainly serine proteinases, such as trypsin/chymotrypsin, and the adult insects only chymotrypsin-like enzymes in their digestive process. Two serine proteinases were separated from zymogram between the 4th and 5th day of larval development and in the adult stage. Effect of soybean trypsin inhibitor (SBTI, a serine proteinase inhibitor) on development of C. capitata was examined by bioassay. C. capitata was susceptible to SBTI which affected larval mass at ED 50 3.01%.

Growth and development of the veliger larvae and juveniles of Polinices pulchellus (Gastropoda: Naticidae)

Adult Polinices pulchellus were collected from the field and held in aquaria under ambient conditions. Egg collars laid by P. pulchellus were cultured at 14°C and 20°C and larval development after hatching was documented photographically. Planktotrophic Polinices pulchellus veligers hatched from egg collars cultured at 20°C after nine to ten days and after 14 to 15 days at 14°C. Veligers spent most of their time close to the water surface and began feeding within one hour of hatching. Repeated attempts to raise larvae to metamorphic competency at 14°C were unsuccessful. Morphological changes, most notably in the colour and size of the velum and foot, were observed in larvae raised at 20°C. During the first 25 days of larval development the velum broadened and bifurcated into four velar arms, the distal regions of which acquired a deep red coloration. By day 40 the foot had increased considerably in size and the degree of black pigmentation. By day 45 pediveligers were competent to metamorphose to the juvenile stage. Exposure to sediment from the adult habitat induced metamorphosis, larvae lost their vela and became benthic juveniles. Within three days of metamorphosis, juvenile snails drilled the bivalve Lasaea adansoni (∼2 mm), later drilled Cerastoderma edule (∼4 mm), and displayed cannibalistic behaviour. Larvae survived for ∼6 months in the absence of a suitable settlement cue.

Embryonic and larval development of Thai Pangas (Pangasius sutchi Fowler, 1937)

The embryonic and larval development of Thai pangas was investigated during peak (May-July 1995) and late spawning (August-October 1995) periods. The fertilized eggs are adhesive and spherical with a yellowish or greenish-brown egg capsule. The yolk sac is yellowish-brown in color and 1.20-1.80 mm in diameter. Nine hours post-fertilization, the first cleavage stage, embryonic shield, head, tail region, neural grooves and somites were evident. The incubation period ranges from 24-36 h at a temperature of 20-30 degrees C. The newly hatched larvae are quite transparent and light yellowish in color with a body length of 2.98-3.10 mm. Eye pigments appear and the heart starts to work within 12-14 h of hatching. In 1-day-old pro-larvae, the mouth becomes well developed; barbules are elongated, prominent and look like tiny threads. The yolk sac is fairly well absorbed and the palatine teeth are fully developed during the 3 day pro-larval stage. At the end of 12 days of larval development, the stomach becomes functional and aerial respiration starts. After 2 weeks, the young fry is well-developed, and is of an adult appearance, that is, measuring up to 13.56 mm in length.