卤虫投喂下长吻(鱼危)仔稚鱼消化酶发育和口宽变化的研究

采用酶学和形态学测定方法, 研究在投喂卤虫条件下长吻(鱼危)仔鱼4种主要消化酶: 胃蛋白酶、胰蛋白酶、脂肪酶和淀粉酶的活性变化以及长吻(鱼危)仔鱼口宽、全长变化。实验共进行13d, 实验结果表明: (1)长吻(鱼危)仔鱼全长、口宽的发育与其日龄表现出明显的线性正相关(R_TL²=0.974, R_MW²=0.964)。口宽与全长比值(MW/TL)在仔鱼开口后急剧下降, 并自7日龄开始维持在0.07—0.08, 口宽和全长处于同步发育期并表现出明显的相关性(R²=0.948), 说明7日龄(/h, days post hatching)后口宽和全长处于同步发育期, 标志仔鱼转食的开始。(2)长吻(鱼危)仔鱼初次开口时即可检测出四种消化酶的活性。5—7/h时胰蛋白酶显著高于初孵仔鱼, 与此时仔鱼开始开口摄食的行为相一致。胃蛋白酶、脂肪酶活性在仔鱼孵化后第7天即开口的第3天, 淀粉酶活性在孵化后第6天, 显著高于初次孵化出来的仔鱼。8—13/h时, 胃蛋白酶、胰蛋白酶、脂肪酶和淀粉酶活性均在较高水平平稳的波动, 标志着消化道发育逐渐健全。     

Ontogeny of the gastrointestinal tract and selected digestive enzymes in cobia Rachycentron canadum (L.)

The ontogeny of the digestive system of cobia Rachycentron canadum from hatching to 22 days post-hatch (dph) (20·1 mm standard length) was examined with light microscopy. The activities of selected pancreatic enzymes were also determined during this period in order to optimize current rearing methods for this species. At hatching (3·6 mm), the digestive tract consisted of a relatively undifferentiated, straight tube positioned dorsally to the yolk sac. The major morphological changes in the digestive tract primarily occurred over the first 1–4 dph (3·6–4·4 mm). During this time, larvae began exogenous feeding (3 dph) and the digestive tract differentiated into five histologically distinct regions: buccopharynx, oesophagus, stomach anlage, anterior intestine and posterior intestine. Yolk reserves were exhausted by 5 dph (4·5 mm) and the oil globule began rapidly decreasing in size disappearing entirely by 9–10 dph (6·3–6·8 mm). Gastric glands differentiated at this time, and by 12 dph (8·1 mm) surface mucous cells of the stomach anlage stained positive for neutral mucosubstances. By 16 dph (11·6 mm), the blind sac (fundic region) of the stomach formed as did the pyloric caecae which initially appeared as a single protrusion of the anterior intestine just ventral to the pyloric sphincter. Generally, enzyme activities (U larva⁻¹) for amylase (0·0–1·8), chymotrypsin (0·0–7902·4), trypsin (0·2–16·6) and lipase (9·3–1319·0) were measurable at or soon after hatching and increased steadily from c. 8–22 dph (5·7–20·1 mm). The results of this study are discussed in terms of current and future weaning practices of this species.     

Histological development of the digestive system of Mayan cichlid Cichlasoma urophthalmus (Günther 1862)

The ontogeny of the digestive tract in Cichlasoma urophthalmus was studied by means of optical microscopy from hatching to 30 days post‐hatching (dph; 855 degree days, dd). The development of the digestive system in this precocial species was a very intense and asynchronous process, which proceeded from both distal ends interiorly. At hatching, the digestive tract consisted of a straight tube with a smooth lumen dorsally attached to the yolk‐sac. The digestive accessory glands were already differentiated and eosinophilic zymogen granules were visible in the exocrine pancreas. At the onset of exogenous feeding between 5 and 6 dph (142.5–171.0 cumulative thermal units, CTU), the buccopharynx, oesophagus, intestine, liver and pancreas were almost completely differentiated, with the exception of the gastric stomach that completed its differentiation between 11 and 14 dph (313.5–399.0 CTU). The development of gastric glands at 14 dph and the differentiation of the stomach in the fundic, cardiac and pyloric regions at 19 dph (541.5 CTU) were the last major events in digestive tract development and designated the onset of the juvenile period. Remnants of yolk were still detected until 16 dph (456.0 CTU), indicating a long period of mixed nutrition that lasted between 10 and 11 days (285.0–313.5 CTU). The results of the organogenesis of larvae complement previous data on the functionality of the digestive system and represent a useful tool for establishing the functional systemic capabilities and physiological requirements of larvae to ensure optimal welfare and growth under aquaculture conditions, which might be useful for improving current larval rearing practices for this cichlid species.     

The ontogenetic development of the digestive tract and accessory glands of sterlet (Acipenser ruthenus L.) larvae during endogenous feeding

The process of differentiation of digestive tract structures in the sterlet Acipenser ruthenus (L.) larvae was studied from hatching to the beginning of exogenous feeding [9 dph (day post hatching)] using histological procedures. On the day of hatching the digestive tract was closed and completely filled with nutrients (the yolk platelets) that were successively utilized during development. A liver primordium was present in the ventral region of the yolksac. The pancreas was observed on the 2 dph. At the same time, the mouth opening took place. Glandular and nonglandular stomach and anterior and intermediate intestine developed from the yolksac walls. Gastric glands became visible on the 7 dph. The primary intestine developed into the spiral intestine. At the moment of onset of exogenous feeding the yolk material was completely exhausted and there was not mixed feeding observed in sterlet larvae. The fish started exogenous feeding on the 9 dph, which was accompanied with evacuation of melanin plug. At the end of endogenous feeding the digestive tract of sterlet larvae was developed and functional, so they could properly utilize food.     

Early development of New Zealand hapuku Polyprion oxygeneios eggs and larvae

This study describes for the first time the normal development of New Zealand hapuku Polyprion oxygeneios embryos and larvae reared from fertilization to 11 days post-hatch (dph) at a constant temperature. Fertilized eggs were obtained from natural spawnings from communally reared captive wild broodstock. Eggs averaged 2 mm in diameter and had single or multiple oil globules. Embryos developed following the main fish embryological stages and required an average of 1859·50 degree hours post-fertilization (dhpf) to hatch. The newly hatched larvae (4·86 mm mean total length, L(T) ) were undifferentiated, with unpigmented eyes, a single and simple alimentary tube and a finfold that covered the entire body. Larvae relied on the energy from the yolk-sac reserves until 11 dph (7·33 mm mean L(T) ), when yolk-sac reabsorption was almost completed. Some of the major developmental stages from hatching to yolk-sac reabsorption were eye pigmentation (5 dph), upper jaw formation (7 dph), lower jaw formation (8 dph) and mouth opening (8-9 dph). By 9 dph, the digestive system consisted of pancreas, liver, primordial stomach, anterior and posterior gut; therefore, P. oxygeneios larvae would be capable of feeding on live prey. The developmental, morphological and histological data described constitutes essential baseline information on P. oxygeneios biology and normal development.     

Ontogeny of gene expression of group IB phospholipase A₂ isoforms in the red sea bream, Pagrus (Chrysophrys) major

The red sea bream (Pagrus major) was previously found to express mRNAs for two group IB phospholipase A₂ (PLA₂) isoforms, DE-1 and DE-2, in the digestive organs, including the hepatopancreas, pyloric caeca, and intestine. To characterize the ontogeny of the digestive function of these PLA₂s, the present study investigated the localization and expression of DE-1 and DE-2 PLA₂ genes in red sea bream larvae/juveniles and immature adults, by in situ hybridization. In the adults, DE-1 PLA₂ mRNA was expressed in pancreatic acinar cells. By contrast, DE-2 PLA₂ mRNA was detected not only in digestive tissues, such as pancreatic acinar cells, gastric glands of the stomach, epithelial cells of the pyloric caeca, and intestinal epithelial cells, but also in non-digestive ones, including cardiac and lateral muscle fibers and the cytoplasm of the oocytes. In the larvae, both DE-1 and DE-2 PLA₂ mRNAs first appeared in pancreatic tissues at 3 days post-hatching (dph) and in intestinal tissue at 1 dph, and expression levels for both gradually increased after this point. In the juvenile stage at 32 dph, DE-1 PLA₂ mRNA was highly expressed in pancreatic tissue, and DE-2 PLA₂ mRNA was detected in almost all digestive tissues, including pancreatic tissue, gastric glands, pyloric caeca, and intestine, including the myomere of the lateral muscles. In conclusion, both DE-1 and DE-2 PLA₂ mRNAs are already expressed in the digestive organs of red sea bream larvae before first feeding, and larvae will synthesize both DE-1 and DE-2 PLA₂ proteins.     

Distribution of cholecystokinin-immunoreactive cells in the gut of developing Atlantic cod Gadus morhua L. larvae fed zooplankton or rotifers

One of the main gastrointestinal hormones, cholecystokinin (CCK), was studied in order to advance understanding of the control of the digestive process in Atlantic cod Gadus morhua larvae after onset of first feeding. Larvae were fed either natural zooplankton or enriched rotifers in similar rearing systems and sampled from hatching to 22 days post-hatch (dph). CCK was visualized by immunohistochemistry and the first CCK-immunoreactive (IR) cells were detected at 8 dph corresponding to 6 days after first feeding. The CCK-IR cells were mostly found in the anterior midgut, and the number of CCK-IR cells was lower in the posterior midgut. They were also present in the hindgut of some of the larvae, but not in the foregut. No clear differences were found in the ontogenetic appearance and the distribution pattern of CCK-IR cells between the two dietary treatments. This indicates that the onset of CCK production in the gut as well as the spatial distribution of the CCK-IR cells is not differentially affected by these diets. To what extent the hormone production itself is influenced by dietary factors needs to be studied by more sensitive methods.     

Red porgy (Pagrus pagrus) larval feeding performance and behavior at the onset of exogenous feeding

The feeding performance and behavior at the onset of exogenous feeding, 3 to 4 days after hatching (DAH), were studied in red porgy Pagrus pagrus larvae. Similar feeding efficiency and intensity were achieved for two feeding treatments (live or freeze-dried rotifers) suggesting that prey movement is not decisive for their detection and capture and demonstrating that at first feeding red porgy larvae can ingest inert food. Larvae feeding performance was not affected by a diet shift between treatments. Based on maximum rotifers consumption and gut evacuation time at 18 °C, the daily ration was estimated as 14.035 μg, considering 14 h of feeding and a 25% egg:female rotifer ratio. Larval swimming activity measured by video recording showed a close association with gut fullness and similar swimming patterns for 3 and 4 DAH larvae. However, 20.3% larger mouth gape and 54.6% higher swimming speed of the older larvae should provide a better feeding performance and more energy needed for growth.     

Morphological development and allometric growth patterns in hatchery-reared California halibut larvae

Morphological development, allometric growth and behaviour of hatchery-reared California halibut Paralichthys californicus were studied from hatching to metamorphosis (42 days post hatch, dph) at 187° C. Mean standard length ( L _S) of larvae and juveniles increased from 2.1 mm at hatching to 10.5 mm at metamorphosis with the increase in length being approximately linear. Stages of morphological development were described using the alphabetic staging (A–I) used for other flatfish species. Organogenesis and differentiation were more rapid and complex in yolk-sac (hatching, stage A–3 dph, stage B), preflexion (3–19 dph, stages B–C), and flexion larvae (from 20 to 23 dph, stages D–E), as larvae developed most of their sensory, feeding, respiratory and swimming systems. After notochord flexion at 24–25 dph (stage F), most morphological changes were related to the progressive transformation from a bilateral symmetrical larva to an asymmetrical benthic juvenile (42 dph, stages G–I).     

Assessment of digestive enzymes activity during the fry development of the endangered Caspian brown trout Salmo caspius

The study of digestive enzymes activity at Salmo caspius fry showed that enzymes were available at the moment of mouth opening on the first day post hatching (dph) and the activity of enzymes showed no significant difference from the hatching day 28 dph. An increased activity was seen between 32 and 43 dph and this activity was significantly higher than the activity during the first 28 days. In the primary stages after yolk sac resorption (43–58 dph), enzymes activity showed an increased profile, however none of them showed a significant difference between 43 and 58 dph.     

Further research on the production, longevity and infectivity of the zoospores of Leptolegnia chapmanii Seymour (Oomycota: Peronosporomycetes)

The effect of temperature on the production, survival and infectivity of zoospores of an Argentinean isolate of Leptolegnia chapmanii was determined under laboratory conditions. Production of zoospores of L. chapmaniiin vitro and in vivo upon first and fourth instars larvae of the mosquito Aedes aegypti was studied at three different temperatures. Zoospores from infected larvae were infective to mosquito larvae for 51, 12, and 5 consecutive days when maintained at 25, 35, and 10 °C, respectively. Maximum zoospore production in infected fourth-instar larvae was 9.6 ± 1.4 × 10⁴ zoosp/larva at 48 h at 25 °C. The average number of zoospores produced by individual fourth-instar Ae. aegypti larvae infected with L. chapmanii was 3.57 ± 0.46 × 10⁵ zoospores during 6 consecutive days at 25 °C. Zoospore production in vitro was also affected by temperature with a maximum of zoospores (n = 47,666/ml) produced at 25 °C. When zoospores produced in vitro were used as inoculum against Ae. aegypti larvae at 25 °C, larval mortality was recorded for 5 consecutive weeks. The encystment process for zoospores took 17-20 min; the germination of cysts (excystment) occurred 5 min after exposure in water to mosquito larvae. The minimal time of contact between zoospores and mosquito larvae to develop infection was two minutes. Infection took place by zoospore attachment onto and then penetration through the larval cuticle or by ingestion of cysts as was confirmed by histological studies. Temperature directly affected infectivity and production of zoospores in vivo and in vitro although L. chapmanii zoospores tolerate a wide range of temperatures.     

Comparative larval ontogeny of two fish species (Characiformes and Siluriformes) endemic to the São Francisco River in Brazil

The aim of the present study was to perform comparative histological analyses of the ontogenetic development of two fish species endemic to the São Francisco River in Brazil: Prochilodus argenteus and Lophiosilurus alexandri. Histological analyses were performed every 24 h from the moment of hatching until 14 days post-hatching (dph) for the observation of larval development and until 39 dph for the observation of gonadal development. Whole larvae were fixed in Bouin's solution and the histological slides were stained with haematoxylin–eosin. Lophiosilurus alexandri larvae had a larger body size compared with P. argenteus larvae since hatching. Lophiosilurus alexandri larvae had mouth opening and pigmentation of the eyes upon hatching, whereas these events were observed at 1 dph in P. argenteus larvae. The visualisation and the inflation of the swim bladder occurred at 1 and 3 dph, respectively, in the P. argenteus, whereas these events occurred at 2 and 8 dph, respectively, in L. alexandri. Yolk granules were absorbed at 4 dph in P. argenteus and the 10 dph in L. alexandri. At 7 dph, the digestive tube was more differentiated in L. alexandri than P. argenteus and at 14 dph, the digestive system of both species had features of their eating habits: broad stomach and short intestine in L. alexandri, typical of carnivorous habits; stomach with a mechanical function and long intestine in P. argenteus, typical of detritivorous habits. The epithelial lining tissue, formed by a single layer of cells in the newly hatched larvae (0 dph), differentiated throughout the study, exhibiting scales in P. argenteus and numerous club cells in the middle epithelial region of L. alexandri at 39 dph. Undifferentiated gonads with somatic cells and primordial germ cells were observed at 39 dph, with caudal-cranial migration since 1 dph in both species. The anatomic changes during the ontogeny of P. argenteus and L. alexandri larvae are directly associated with the evolutionary history of each species, which explains their feeding habits, behaviour and distribution in the environment: Prochilodus argenteus is detritivorous and actively swims in the water column, whereas L. alexandri is carnivorous and inhabits bottom regions. At 39 dph neither species exhibited sexual differentiation.     

A histological study of the organogenesis of the digestive system in bay snook Petenia splendida Günther, 1862 from hatching to the juvenile stage

In bay snook (Petenia splendida) larvae the histological development of the digestive system and swim bladder, and their relative timing of differentiation were studied from hatching to 45 days post‐hatch (dph) at 29°C. Newly hatched larvae showed a simple digestive tract, which appeared as a straight undifferentiated tube lined by a single layer of columnar epithelial cells (future enterocytes). The anatomical and histological differentiation of the digestive tract and accessory glands was a very intense, asynchronous process, proceeding from the distal to the anterior part. The intestine was the first region to differentiate (9 days post‐hatch – dph, 6.5 mm SL), and the oesophagus the last (21 dph, 8.4 mm SL). At the onset of feeding, the digestive system was organized into different functional and histologically differentiated sections, such as the buccopharynx, oesophagus, glandular stomach, and anterior and posterior intestine. This organization resembled that of the juveniles, with the exception of pharyngeal teeth and buccopharyngeal as well as oesophageal goblet cells, which proliferated later during the mixed feeding period. Histological observations revealed that bay snook larvae retained endogenous yolk reserves until 24 dph (8.9 ± 0.4 mm SL), which might be helpful for weaning this species onto a compound diet. The important lipidic accumulation observed in the intestinal mucosa, liver, and pancreas in fish fed a compound trout diet indicated that although fish were able to digest and absorb lipids, the diet formulation did not fit the nutritional requirements of early juveniles of this species. The ontogeny of the digestive system followed the same general pattern as in most cichlid species described to date. However, we detected species‐specific differences in the timing of differentiation that were related to their reproductive guild. According to the histological results, some recommendations regarding the intensive culture of this species are also provided.     

Cost of oviposition site selection in a water strider Aquarius paludum insularis: Egg mortality increases with oviposition depth

Females generally avoid selecting sites for oviposition which have a high predation risk to increase offspring survival. Previous studies have focused on costs to ovipositing females. However, although offspring may also incur costs by being oviposited at low predation risk sites, no studies have focused on costs to offspring. Such costs to offspring were examined by using Aquarius paludum insularis, females of which avoid eggs parasitism by ovipositing at deep sites. Deep sites are safe from egg parasitism but may be unsuitable for hatching due to environmental factors. We examined the costs to offspring at deep sites by comparing the hatching rate, the duration to hatching and the proportion of drowned larvae between eggs that were set at three levels of water depth (0 cm, 25 cm and 50 cm depth). While the hatching rate at 50 cm was lower than that at 0 cm, the rate at 25 cm did not differ from that at 0 cm. Duration to hatching and the proportion of drowned larvae did not differ between the three depths. It is suggested that the declining survival rate of A. paludum eggs was due to increased water pressure at greater depth. Such a cost may exist in other species and such an observation may aid in understanding oviposition site selection.     

Embryo and larva development in common dentex (Dentex dentex), a pelagophil teleost: The quantitative composition of egg-free amino acids and their interrelations

Free amino acids (FAAs) play a key role in the physiology of marine teleosts (eggs, embryos, and larvae). However, the relationship between the egg FAAs content and the production of viable embryos and larvae (at different developmental stages) in batch spawner pelagophils has not yet comprehensively been investigated. Viable eggs of common dentex, Dentex dentex, were obtained from captive broodstocks. Egg wet weight (WW), dry weight (DW), and water content (%W) and viability parameters, or VPs (egg floating rate [FR], hatching rate [HR], and larval survival rate [SR] at days 0 to 5 posthatch) were determined for 45 egg batches. The egg batches were classified according to their HR magnitude. Twelve egg batches with the same WW, DW, and %W were taken from the same broodstock and at the same developmental stage to determine the qualitative and quantitative composition of FAAs. The total FAA (TFAA) content, glutamic acid (Glu), asparagine (Asn), glutamine (Gln), and arginine (Arg) were correlated with VPs. The Glu was significantly correlated with HR and SR at 0 day posthatch (dph), the Asn with SR at 1 dph, and the Gln and Arg with FR and HR. Of the 361 ratios made based on the absolute concentrations of FAAs, 24 ratios were correlated with VPs (P < 0.005) through 42 simple regression models (R² = 0.641 to 0.846). Of the 42 significant relationships found ∼10%, ∼28%, ∼12%, ∼30%, ∼8%, ∼4%, ∼2%, ∼2%, and ∼2% of the models show the relations of the egg FAAs ratios with FR, HR, SR at days 1 to 5 posthatch, and %W, respectively. A path coefficient in combination with a Pearson's correlation coefficient provided a series of statistical evidences to show the effects of the egg FAAs interrelations on the relationships found between quantitative composition of a FAA and a VP.     

Daily rhythms of clock gene expression and feeding behavior during the larval development in gilthead seabream,Sparus aurata

Light is the main environmental time cue which synchronizes daily rhythms and the molecular clock of vertebrates. Indeed, alterations in photoperiod have profound physiological effects in fish (e.g. reproduction and early development). In order to identify the changes in clock genes expression in gilthead seabream larvae during ontogeny, three different photoperiods were tested: a regular 12L:12D cycle (LD), a continuous light 24L:0D (LL) and a two-phases photoperiod (LL?+?LD) in which the photoperiod changed from LL to LD on day 15 after hatching (dph). Larvae were sampled on 10, 18, 30 and 60 days post-hatch (dph) during a 24?h cycle. In addition to the expression of clock genes (clock, bmal1, cry1 and per3), food intake was measured. Under LD photoperiod, larvae feed intake and clock genes expression showed a rhythmic pattern with a strong light synchronization, with the acrophases occurring at the same hour in all tested ages. Under LL photoperiod, the larvae also showed a rhythmic pattern but the acrophases occurred at different times depending on the age, although at the end of the experiment (60 dph) clock genes expression and feed intake rhythms were similar to those larvae exposed to LD photoperiod. Moreover, the expression levels of bmal1 and cry1 were much lower than in LD photoperiod. Under the LL?+?LD photoperiod, the 10 dph larvae showed the same patterns as LL treatment while 18 and 30 dph larvae showed the same patterns as LD treatment. These results revealed the presence of internal factors driving rhythmic physiological responses during larvae development under constant environmental conditions. The LL?+?LD treatment demonstrates the plasticity of the clock genes expression and the strong effect of light as synchronizer in developing fish larvae.