Identification and expression analysis of transcripts involved in taurine biosynthesis during early ontogeny of tropical gar Atractosteus tropicus.

In fishes, the availability of taurine is regulated during ontogenetic development, where its endogenous synthesis capacity is species dependent. Thus, different pathways and involved enzymes have been described: pathway I (cysteine sulfinate-dependent pathway), cysteine dioxygenase type 1 (cdo1) and cysteine sulfinic acid decarboxylase (csad); pathway II (cysteic acid pathway), cdo1 and glutamic acid decarboxylase (gad); and pathway III (cysteamine pathway), 2-aminoethanethiol dioxygenase (ado); whereas taurine transporter (taut) is responsible for taurine entry into cells on the cell membrane and the mitochondria. This study determined if the tropical gar (Atractosteus tropicus), an ancient holostean fish model, has the molecular mechanism to synthesize taurine through the identification and analysis expression of transcripts coding for proteins involved in its biosynthesis and transportation, at different embryo-larvae stages and in different organs of juveniles (31 dah). We observed a fluctuating expression of all transcripts involved in the three pathways at all analyzed stages. All transcripts are expressed during the beginning of larval development; however, ado and taut show a peak expression at 9 dah, and all transcripts but csad decreased at 23 dah, when the organism ended the larval period. Furthermore, at 31 dah, we observed taut expression in all examined organs. The transcripts involved in pathways I and III are expressed differently across all organs, whereas pathway II was only observed in the brain, eye, and skin. The results suggested that taurine biosynthesis in tropical gar is regulated during its early development before first feeding, and the pathway might also be organ-type dependent.

Morphological and cardiac alterations after crude oil exposure in the early-life stages of the tropical gar (Atractosteus tropicus).

Fish development can be affected by environmental pollutants such as crude oil (anthropogenic or natural sources), causing alterations especially in cardiac function and morphology. Most such studies have focused on saltwater species, whereas studies in freshwater fishes are scant. The objective of the current study was to evaluate the effects of crude oil exposure (as 0, 5, 10, 15, or 20% high-energy water accommodated fractions, HEWAF) on cardiac function and edema formation during two early periods of development (embryo and eleuteroembryo, 48 h each) individually using the tropical gar Atractosteus tropicus as a model. Embryos did not exhibit alterations in body mass, total length, condition factor, and cardiac function as a function of oil. In contrast, eleuteroembryos proved to be more sensitive and exhibited increased body mass, total length, and condition factor, decreased heart rate and phenotypic alterations such as cardiac dysmorphia (tubular hearts) and spine curvature at high concentrations of HEWAF. Moreover, edema formation was observed in both stages This study shows different functional responses of A. tropicus after crude oil exposure and provides useful information of the developmental impacts of these compounds on the early life stages of freshwater tropical fishes.

Hypoxia- and hyperoxia-related gene expression dynamics during developmental critical windows of the tropical gar Atractosteus tropicus.

Aquatic hypoxia is both a naturally-occurring and anthropogenically-generated event. Fish species have evolved different adaptations to cope with hypoxic environments, including gill modifications and air breathing. However, little is known about the molecular mechanisms involved in the respiration of embryonic and larval fishes during critical windows of development. We assessed expression of the genes hif-1α, fih-1, nhe1, epo, gr and il8 using the developing tropical gar as a piscine model during three developmental periods (fertilization to hatch, 1 to 6 days post hatch (dph) and 7 to 12 dph) when exposed to normoxia (~7.43 mg/L DO), hypoxia (~2.5 mg/L DO) or hyperoxia (~9.15 mg/L DO). All genes had higher expression when fish were exposed to either hypoxia or hyperoxia during the first two developmental periods. However, fish continuously exposed to hypoxia had increased expression of the six genes by hatching and 6 dph, and by 12 dph only hif-1α still had increased expression. The middle developmental period was the most hypoxia-sensitive, coinciding with several changes in physiology and morphology. The oldest larvae were the most resilient to gene expression change, with little variation in expression of the six genes compared. This study is the first to relate the molecular response of an air-breathing fish to oxygen availability to developmental critical windows and contributes to our understanding of some molecular responses of developing fish to changes in oxygen availability.

Changes in digestive enzyme activities during the early ontogeny of the South American cichlid (Cichlasoma dimerus).

Cichlasoma dimerus is a neotropical cichlid that has been used as a biological model for neuroendocrinology studies. However, its culture is problematic in terms of larval feeding to allow having enough fry quantity and quality. Larviculture requires full knowledge about the digestive system and nutrition; therefore, this study was intended to assess the digestive enzymes' changes at different ages during the early ontogeny. Acid protease activity was detectable from the first day after hatching (dah), increasing to its maximum peaks on 9 dah. In contrast, alkaline proteases had low activity in the first days of life but reached their maximum activity on 17 dah. Chymotrypsin, L-aminopeptidase, and carboxypeptidase A activities increased at 6 dah, while trypsin activity was first detected on 13 dah and reached its maximum activity on 17 dah. Lipase and α-amylase activity were detectable at low levels in the first days of life, but the activity fluctuated and reaching its maximum activity at 21 dah. Alkaline phosphatase continued to oscillate and had two maximum activity peaks, the first at 6 dah and the second at 19 dah. Zymograms of alkaline proteases on day 6 dah six revealed four activity bands with molecular weights from 16.1 to 77.7 kDa. On 13 dah, two more activity bands of 24.4 and 121.9 kDa were detected, having a total of six proteases. The enzymatic activity analyzes indicate the digestive system shows the low activity of some enzymes in the first days after hatching, registering significant increases on 6 dah and the maximum peaks of activities around at 17 dah. Therefore, we recommend replacing live food with dry feed and only providing dry feed after day 17 dah.

Effects of Non-Nutritive Sweeteners on Energy Intake, Body Weight and Postprandial Glycemia in Healthy and with Altered Glycemic Response Rats.

The aim of this study was to evaluate the effects of non-nutritive sweeteners (NNS) consumption on energy intake, body weight and postprandial glycemia in healthy and with altered glycemic response rats. Animals on normal diet (ND) or high-fat diet (HFD) were divided to receive NNS (sucralose, aspartame, stevia, rebaudioside A) or nutritive sweeteners (glucose, sucrose) for 8 weeks. The NNS were administered at doses equivalent to the human acceptable daily intake (ADI). A test using rapidly digestible starch was performed before and after treatments to estimate glycemic response. No effects of NNS consumption were observed on energy intake or body weight. Sucrose provoked an increased fluid consumption, however, energy intake, and weight gain were not altered. In ND, no effects of NNS on glycemic response were observed. In HFD, the glycemic response was increased after sucralose and stevia when only the final tolerance test was considered, however, after including the baseline test, these results were no longer significant compared to glucose. These findings provide further evidence suggesting that at the recommended doses, NNS do not alter feeding behavior, body weight or glycemic tolerance in healthy and with altered glycemic rats.

Comparative characterization of digestive proteases in redhead cichlid (Vieja melanurus) and twoband cichlid (Vieja bifasciata) (Percoidei: Cichlidae)

In the Southeast of Mexico, there are many native cichlids with commercial interest such as redhead cichlid (Vieja melanurus) and twoband cichlid (V. bifasciata), which have a great local demand and excellent meat quality. However, it is necessary to implement their culture based on nutrition studies and digestive biochemistry. This study's objective was to characterize these two cichlids' digestive proteases (pH, temperature, and inhibitors) through biochemistry techniques. Results showed that V. melanurus and V. bifasciata have a digestive capacity analogous to other omnivore fishes, where the optimal pH values of stomach proteases (4 and 2, respectively) and intestinal proteases (6 and 12, respectively), the optimal temperature of acid (35°C and 55°C, respectively) and alkaline proteases (45°C and 55°C, respectively) are quite similar. Both species presented high thermal and pH stabilities. Inhibition showed that V. melanurus is more sensitive to specific inhibitors for alkaline proteases than V. bifasciata. In conclusion, V. bisfasciata and V. melanurus have different digestive protease patterns. Both species can hydrolyze different protein ingredients to formulate a specific diet. Nevertheless, V. bifasciata is more resistant to the presence of inhibitors, which allow it to include vegetable proteins in its diet.(AU)

En el sureste de México, existen muchas especies de cíclidos nativos de interés comercial como el cíclido rojo (Vieja melanurus) y el cíclido de dos bandas (V. bifasciata), los cuales tienen una gran demanda local y tienen una excelente calidad de carne; sin embargo, es necesario implementar su cultivo con base en estudios de nutrición y bioquímica digestiva. El objetivo de este estudio fue caracterizar las proteasas digestivas (pH, temperatura e inhibidores) de estos dos cíclidos nativos mediante técnicas bioquímicas. Los resultados mostraron que V. melanurus y V. bifasciata tienen una capacidad digestiva similar a otros peces omnívoros, donde los valores óptimos de pH de proteasas estomacales (4 y 2, respectivamente) e intestinales (6 y 12, respectivamente), la temperatura óptima de proteasas ácidas (35°C y 55°C, respectivamente) y alcalinas (45°C y 55°C, respectivamente) son muy parecidas. Ambas especies presentaron alta estabilidad térmica y de pH. La inhibición mostró que V. melanurus es más sensible a inhibidores específicos de proteasas alcalinas que V. bifasciata. En conclusión, V. bisfasciata y V. melanurus tienen diferentes patrones de proteasas digestivas, pero ambas especies pueden hidrolizar diversos ingredientes proteicos para formular dietas específicas; sin embargo, V. bifasciata es más resistente a la presencia de inhibidores, lo que permitiría incluir proteínas vegetales en su dieta.(AU)

Continuous Microalgal Cultivation for Antioxidants Production.

Microalgae, including cyanobacteria, represent a valuable source of natural compounds that have remarkable bioactive properties. Each microalga species produces a mixture of antioxidants with different amounts of each compound. Three aspects are important in the production of bioactive compounds: the microalga species, the medium composition including light supplied and the photobioreactor design, and operation characteristics. In this study, the antioxidant content and productivity performance of four microalgae were assessed in batch and continuous cultures. Biomass productivity by the four microalgae was substantially enhanced under continuous cultivation by 5.9 to 6.3 times in comparison with batch cultures. The energetic yield, under the experimental conditions studied, ranged from 0.03 to 0.041 g biomass kJ-1. Phenols, terpenoids, and alkaloids were produced by Spirulinaplatensis, Isochrysisgalbana, and Tetraselmissuecica, whereas tocopherols and carotenoids were produced by the four microalgae, except for phycocyanin and allophycocyanin, which were only produced by S. platensis and Porphyridiumcruentum. The findings demonstrate that the continuous cultivation of microalgae in photobioreactors is a convenient method of efficiently producing antioxidants.

Identification and in silico structural and functional analysis of a trypsin-like protease from shrimp Macrobrachium carcinus.

Macrobrachium carcinus (Linnaeus, 1758) is a species of freshwater shrimp widely distributed from Florida southwards to southern Brazil, including southeast of Mexico. In the present work, we identified a putative trypsin-like protease cDNA fragment of 736 nucleotides from M. carcinus hepatopancreas tissue by the 3'RACE technique and compared the deduced amino acid sequence to other trypsin-related proteases to describe its structure and function relationship. The bioinformatics analyses showed that the deduced amino acid sequence likely corresponds to a trypsin-like protease closely related to brachyurins, which comprise a subset of serine proteases with collagenolytic activity found in crabs and other crustacea. The M. carcinus trypsin-like protease sequence showed a global sequence identity of 94% with an unpublished trypsin from Macrobrachium rosenbergii (GenBank accession no. AMQ98968), and only 57% with Penaeus vannamei trypsin (GenBank accession no. CAA60129). A detailed analysis of the amino acid sequence revealed specific differences with crustacean trypsins, such as the sequence motif at the beginning of the mature protein, activation mechanism of the corresponding zymogen, amino acid residues of the catalytic triad and residues responsible for substrate specificity.

Effect of salinity on zinc toxicity (ZnCl2 and ZnO nanomaterials) in the mosquitofish (Gambusia sexradiata).

Zn is an essential trace metal in living beings. However, excessive concentrations can cause toxic effects even in the aquatic biota. Zn is widely used in different industrial sectors, which has increased its presence in aquatic environments. To assess the acute toxicity of Zn, bioassays were performed with the fish Gambusia sexradiata for a 96-h exposure using ZnCl2 (0 and 15 salinity) and ZnO nanomaterials (0 salinity). The mean lethal concentrations (LC50-96 h) for ZnCl2 were 25.36 (19.64-32.76) and 177.91 (129.39-244.63) mg Zn L-1 to 0 and 15 salinity, respectively. The increased concentration of ZnCl2 showed a dose-response relationship; similarly, the increase in salinity significantly reduces the toxicity of Zn. Characterisation of ZnO nanomaterials was carried out by FTIR, DRX, SEM, DLS and zeta potential. The FTIR spectra showed the characteristic band of Zn-O vibration at 364 cm-1, while DRX presents the hexagonal wurtzite structure with an average crystallite size of 40 nm. SEM micrographs reveal rod-like shapes with lengths and diameters of 40-350 nm and 90 nm, respectively. Agglomerates of 423 nm in water suspension were obtained by DLS and zeta potential of + 14.4 mV. Under these conditions, no mortality was observed due to the rapid flocculation/precipitation of ZnO nanomaterials, which involved brief interaction periods of Zn in the water column with the fish. Gambusia sexradiata is affected by increased Zn concentrations in hard water conditions, and salinity changes modified Zn toxicity, placing it as a suitable model for toxicity tests for this type of particles.

Trypsin gene expression in adults and larvae of tropical gar Atractosteus tropicus.

Trypsin gene (try) expression levels were quantified in different organs of wild and captive tropical gar (Atractosteus tropicus) adults, and changes in expression during initial ontogeny of the species were determined. RNA was extracted from the pancreas, and cDNA was synthesized and later amplified by endpoint PCR using oligonucleotides designed from different try sequences of fish registered in GenBank. Subsequently, specific oligonucleotides were designed from the partial sequences. Gene expression was measured after RNA extraction and synthesis of the cDNA of 11 organs (liver, pancreas, stomach, esophagus, intestine, pyloric caeca, brain, muscle, gills, gonad, and kidney) of captive and wild adults. Likewise, samples of A. tropicus larvae were taken on days 0 (embryo), 5, 10, 15, 20, 25, and 30 days after hatching (DAH), the RNA was extracted, and the synthesis of cDNA was carried out to measure real-time gene expression (qPCR). The results showed that the highest relative try expression occurred mainly in the esophagus, liver, stomach, and pancreas of both wild and captive adult fish; however, captive organisms had a higher try expression level than wild fish. Although try expression during initial ontogeny was high in embryos (0 DAH), it did not reach the maximum value until 15 DAH. It was concluded that try expression levels in captive adults are due to the high protein content in the balanced feed (trout diet). The highest try expression level during larviculture was detected at 15 DAH, which indicates that A. tropicus larvae have a mature digestive system and can efficiently hydrolyze proteins from feed at this developmental stage.

Biochemical characterization of a semi-purified aspartic protease from sea catfish Bagre panamensis with milk-clotting activity.

Pepsin from stomach of Bagre panamensis was semi-purified and biochemically characterized. The acid proteolytic activity and purification fold were 3875 U/mg protein and 91.85, respectively, after purification process. The optimum pH and temperature for semi-purified protease were 2-3 and 65 °C, respectively. The enzyme activity was stable after heating proteases at 50 °C for 120 min, but only 30% residual activity was detected after heating at 65 °C for 30 min. SDS-PAGE analysis showed two proteins bands after dialysis (26.1 and 38.6 kDa). Only the band of 38.6 kDa had proteolytic activity, which was inhibited using pepstatin A. Organic solvents, surfactants and reducing agents affect the proteolytic activity at different extent; however, metal ions or EDTA have no impact on protease activity. The semi-purified protease exhibited milk coagulant activity, with a maximum activity at 45 °C. The obtained results highlight the potential biotechnological use of B. panamensis pepsin.

Changes in digestive enzyme activities during early ontogeny of Seriola rivoliana.

This study contributes to basic knowledge on the development of larval digestive enzyme activities of Seriola rivoliana, a carnivorous species that has shown high mortality in larval stage. The study was conducted under laboratory-controlled conditions until larvae reached 30 days after hatching (DAH). The specific activity of amylase and pepsin was analysed by spectrophotometric methods, and lipase, trypsin, chymotrypsin and alkaline phosphatase by fluorometry. The results showed that S. rivoliana larvae had different enzyme patterns depending on their developmental stage, and also when exogenous food was administered (rotifers and Artemia). At hatching (2.98 ± 0.14 mm SL), lipase, alkaline phosphatase, trypsin and chymotrypsin activities were detected, but the last two enzymes showed an activity close to zero. Trypsin, chymotrypsin, lipase and alkaline phosphatase sharply increased their activities after starting the first feeding. Amylase activity was not detected until 5 DAH (3.51 ± 0.18 mm SL) and had a peak on day 15 (4.59 ± 0.39 mm SL); no activity was observed from then on. Pepsin activity was detected approximately at 15 DAH, showing a constant increase onwards. Trypsin had a peak on day 15 and then decreased on the following days. Chymotrypsin had the highest peak at 5 DAH, then a sharp decrease on day seven (3.88 ± 0.11 mm SL), and it increased gradually until day 30 (11.71 ± 1.59 mm SL). Lipase showed the highest activity at 15 DAH and then decreased its value. Finally, alkaline phosphatase showed the highest value at 3 DAH (3.62 ± 0.06 mm SL); then, a severe decrease was observed with activity stability on the following days. In conclusion, S. rivoliana larvae had a similar pattern as that observed in other marine-carnivorous fish larvae, so it could be considered as juvenile at 20 DAH in terms of digestive physiology; based on our results and previous study, the weaning process could start after this age, which would allow establishing a co-feeding protocol using inert diet.

Changes in digestive enzymes activities during the initial ontogeny of wolf cichlid, Parachromis dovii (Perciformes: Cichlidae)

Wolf cichlid, Parachromis dovii, is a species with a high potential for aquaculture in Central America; however, the knowledge of the digestive physiology in larvae period is limited. For these reason, this study evaluated the changes on digestive enzymes (alkaline and acid proteases, trypsin, chymotrypsin, aminopeptidase, carboxypeptidase, lipases, amylases, and phosphatases) during early ontogeny by biochemical analysis. All digestive enzymes were detected at first feeding (6 days after hatching, DAH, 9.49 mm, 168 degree-days DD). Afterwards all enzymes reached two main peaks in activity at 14 or 22 DAH (15.10 mm, 364 DD and 20.83 mm, 550 DD, respectively). Later, there was a gradual decrease in activity for trypsin and acid and alkaline phosphatases until reach the lowest values at 41 DAH. In the case of acid proteases, chymotrypsin, aminopeptidase, carboxypeptidase, lipase and amylase, all activities reached their maximum values at the end of the larval period, except for alkaline proteases, which showed the maximum value at 14 DAH (15.10 mm, 364 DD). Parachromis dovii larvae have an early capability to hydrolyze exogenous food, agreeing with other carnivorous neotropical cichlid species, for this reason we proposed that the weaning process could begin at 14 DAH.(AU)

El guapote lagunero (Parachromis dovii) es una especie con un alto potencial para la acuicultura en la región de América Central; sin embargo, existe un conocimiento limitado sobre la capacidad digestiva en el periodo larval. Por este motivo, este estudio evaluó los cambios de las enzimas digestivas (proteasas alcalinas y ácidas, tripsina, quimotripsina, aminopeptidasa, carboxipeptidasa, lipasas, amilasas y fosfatasas) durante la ontogenia temprana mediante análisis bioquímico. Todas las enzimas digestivas analizadas se detectaron en la primera alimentación (6 días después de la eclosión, DAH, 9.49 mm, 168 día-grados DD). Después, todas las enzimas alcanzaron dos picos máximos a los 14 o 22 DAH (15.10 mm, 364 DD and 20.83 mm, 550 DD, respectivamente). Después las actividades tripsina, fosfatasas ácidas y alcalina disminuyeron a sus valores más bajos a los 41 DAH. En el caso de las proteasas ácidas y alcalinas, quimotripsina, aminopeptidasa, carboxipeptidasa, lipasa y amilasa, los niveles de actividad aumentaron y alcanzaron su máximo valor al final del período larvario, excepto las proteasas alcalinas, que mostraron su máximo valor a los 14 DAH (15.10 mm, 364 DD). Las larvas de P. dovii tienen una capacidad temprana para hidrolizar alimentos exógenos, lo que concuerda con otras especies de cíclidos neotropicales carnívoros, por lo que proponemos que el proceso de destete inicie a los 14 DAH.(AU)

Alimentos microencapsulados para el cultivo de larvas de pejelagarto (Atractosteus tropicus) / Microencapsulated diets for tropical gar (Atractosteus tropicus) larviculture

Resumen El pejelagarto es una especie importante para la acuicultura en el sureste de México. Se han realizado varios estudios sobre nutrición y fisiología digestiva; sin embargo, el uso de microcápsulas para la alimentación de larvas que permitan mejorar el crecimiento y la supervivencia no ha sido realizado. En este sentido, se evaluaron cuatro alimentos microencapsulados en el crecimiento y supervivencia de larvas de pejelagarto (Atractosteus tropicus). Los tratamientos consistieron en cuatro dietas experimentales y un control: 1) Microcápsula con base en harina de pescado (DP), 2) Microcápsula con base en una combinación de harina de cerdo y pollo (Re), 3) Microcápsula con base en Nannochloropsis gaditana (Ng), 4) Microcápsula con base en un preparado enzimático, pancreatina (PE) el tratamiento control 5) Nauplios de Artemia (Na). Los resultados de las dietas indican que las larvas alimentadas con Nauplios (Na) tuvieron el mayor crecimiento y supervivencia (3.93 cm, 0.19 g y 82 % respectivamente). Mientras que en el caso de las larvas alimentadas con las dietas microencapsuladas, los mejores valores de supervivencia se observaron con las dietas Nannochloropsis gaditana (Ng) y preparado enzimático (PE) (20.0 y 19.2 % respectivamente). Nuestros resultados demuestran que el uso de microencapsulados para la alimentación de larvas de A. tropicus es factible, aunque se requieren más investigaciones para optimizar su diseño y elaboración para mejorar el crecimiento y supervivencia de los organismos.

Abstract Tropical gar is an important species for aquaculture purposes in Southeast Mexico. Several studies regarding nutrition and digestive physiology have been done; however, the use of microcapsules for larvae feeding to improve growth and survival has not been conducted. In this sense, four microencapsulated diets were evaluated with respect to the growth and survival of tropical gar Atractosteus tropicus larvae. The treatments consisted of four experimental diets and one control diet 1) fish meal, 2) a combination of pork and poultry meals, 3) Nannochloropsis gaditana meal, 4) enzyme preparation and the control treatment 5) Artemia nauplii. The evaluation indicated that the larvae fed the Artemia nauplii obtained the greatest growth and survival (3.93 cm, 0.19 g y 82 % respectively), which justifies a correct culture system operation. While larvae fed microencapsulated diets best values in survival were observed with diets Nannochloropsis gaditana and enzyme preparation (20.0 and 19.2 %). Our results showed that microencapsulated could be used to feed A. tropicus in feasibly form. However, more information concerning to optimize the design and manufacturing are required to improve the growth and survival of organisms. Rev. Biol. Trop. 66(3): 1298-1313. Epub 2018 September 01.

Fish trypsins: potential applications in biomedicine and prospects for production.

In fishes, trypsins are adapted to different environmental conditions, and the biochemical and kinetic properties of a broad variety of native isoforms have been studied. Proteolytic enzymes remain in high demand in the detergent, food, and feed industries; however, our analysis of the literature showed that, in the last decade, some fish trypsins have been studied for the synthesis of industrial peptides and for specific biomedical uses as antipathogenic agents against viruses and bacteria, which have been recently patented. In addition, innovative strategies of trypsin administration have been studied to ensure that trypsins retain their properties until they exert their action. Biomedical uses require the production of high-quality enzymes. In this context, the production of recombinant trypsins is an alternative. For this purpose, E. coli-based systems have been tested for the production of fish trypsins; however, P. pastoris-based systems also seem to show great potential in the production of fish trypsins with higher production quality. On the other hand, there is a lack of information regarding the specific structures, biochemical and kinetic properties, and characteristics of trypsins produced using heterologous systems. This review describes the potential uses of fish trypsins in biomedicine and the enzymatic and structural properties of native and recombinant fish trypsins obtained to date, outlining some prospects for their study.

Larval development of the Mexican Snook, Centropomus poeyi (Teleostei: Centropomidae)

We document for the first time the early ontogeny of Centropomus poeyi based on captive raised material representing 0-19 days posthatch (dph). The achievement of early developmental landmarks (i.e., yolk-sac depletion, flexion, development of fins) and changes in pigmentation are described (1.4 mm NL-10.6 mm SL; 0-19 dph) and documented for a subset of individuals using high quality photographs. The ontogeny of the viscerocranium is also described (2.4 mm NL-10.6 mm SL; 6-19 dph). Development in C. poeyi occurs over a short period with attainment of the juvenile stage (i.e., full complement of fin rays present in each fin) occurring by 6.9 mm SL. The ontogeny of external pigmentation in C. poeyi is marked by two trends throughout growth: (1) a decrease in pigmentation dorsally; and (2) an increase in pigmentation ventrally along the midline. Development of the viscerocranium begins with the appearance of the maxilla and dentary in individuals of 2.4 mm NL, coinciding with the depletion of the yolk-sac. By 10.6 mm SL all bones of the viscerocranium are present and teeth are present on all teeth-bearing bones of the adult. Aspects of early development in C. poeyi are compared with the congeners C. undecimalis and C. parallelus.(AU)

Se documentó por primera vez la ontogenia temprana de Centropomus poeyi basada en material obtenido de cautiverio a partir de embriones (0 días después de la eclosión, dde) hasta los 19 dde. Se registró la aparición de puntos de referencia en el desarrollo temprano (p.ej. consumo de saco vitelino, flexión, desarrollo de aletas) y los cambios de pigmentación han sido descritos (1.4 mm NL-10.6 mm SL; 0-19 dde), siendo documentado para un subgrupo de individuos utilizando fotografías de alta calidad. El desarrollo del viscerocráneo también fue descrita (2.4 mm NL-10.6 mm SL; 6-19 dde). El desarrollo en C. poeyi sucede en un periodo corto de tiempo hasta alcanzar una etapa juvenil (p.ej., aparición completa de los radios presentes en cada aleta), lo que se observa a los 6.9 mm SL. La pigmentación externa en C. poeyi esta marcada por dos tendencias en el crecimiento: (1) un descenso en la pigmentación dorsal; y (2) un incremento en la pigmentación ventral a lo largo de la línea media. El desarrollo del viscerocráneo comienza con el aparecimiento del maxilar y el dentario en individuos de 2.4 mm NL, lo cual coincide con el consumo del saco vitelino. A los 10.6 mm SL todos los huesos del viscerocráneo están presentes como se observa en los dientes en los adultos. Algunos aspectos del desarrollo temprano en C. poeyi son comparados con los congéneros C. undecimalis y C. parallelus.(AU)

Histological study of the gastrointestinal tract in longfin yellowtail (Seriola rivoliana) larvae.

This work contributes basic knowledge on larval development of Seriola rivoliana. A histological study describes the development of the digestive tract and accessory glands in S. rivoliana larvae reared under laboratory conditions at 24 °C from hatching to 30 days post-hatching (DPH). At hatching (2.6 ± 0.12 mm), larvae had an undifferentiated digestive tract with a closed straight tube and a large yolk sac with an oil globule. The liver and pancreas were observed at 1 and 2 days, and the mouth and anus opened at day 2. Enriched rotifers were visible in their digestive tract. At the beginning of the pre-flexion stage, a mixed nutritional period was observed. At day 3, exogenous feeding began; the digestive tract became differentiated into the buccopharynx, esophagus, an undifferentiated stomach, and the intestines. Zymogen granules were visible in the exocrine pancreas. At day 4, supranuclear vacuoles were present in the posterior intestine, indicating the beginning of intracellular digestion. At day 5, goblet cells were present in the esophagus and became functional at day 7 in the esophagus and intestine. The buccopharynx goblet cells developed at day 15. The presence of gastric glands and differentiation of the stomach in the fundic, cardiac, and pyloric regions during the post-flexion stage occurred at day 20. This was the onset of the juvenile period and the beginning of weaning; however, a long co-feeding phase is recommended. Pyloric caeca were observed at day 30 (13.6 ± 1.6 mm). These results provide valuable information on S. rivoliana larvae biology and digestive physiology, which should be useful to improve cultivation techniques and identify ecological features involved in ontogeny.

Permanent germinal epithelium and reproductive cycle of Atractosteus tropicus (Lepisosteiformes: Lepisosteidae) males, Tabasco, Mexico.

The tropical gar A. tropicus plays an important ecological role as it regulates other fish stocks in different water bodies in Southeastern México. Nevertheless, wild populations are declining, and one conservation alternative is the aquaculture production and basic knowledge of reproductive biology; for males, this requires the study of germ and somatic structures of testes, to characterize the reproductive cycle, and to provide basic knowledge for exploitation and conservation models and strategies. With this aim, a total of 24 males with an average Lp = 47.2 cm were collected from wild populations from the Laguna Pomposú, municipality of Jalpa de Mendez (18°19' - 93°01'12" W), Tabasco, Mexico. Fish were collected with a trawl net and were transported live to the Tropical Aquaculture Laboratory, División Académica de Ciencias Biológicas (DACBiol), Universidad Juárez Autónoma de Tabasco (UJAT). Males were slaughtered by prolonged immersion in MS222. Testes samples were collected from each specimen and were processed using the standard histological procedures, that consisted of dehydration in an ascending ethanol series, xylol, embedding in paraffin, sectioning at 7 µm, and staining with hematoxylin-eosin (HE). The diameter of 20 seminiferous tubules (Tse), height of germinal epithelium (Egl), gonadosomatic index (IGS) and gonad volume (gV) were determined monthly. Based on morphometric and morpho-physiological characteristics, the testes consisted of a network of anastomosed tubules with non-restricted cystic spermatogenesis, and a permanent germinal epithelium. This is the first report of a permanent germinal epithelium in A. tropicus. Five reproductive classes were histologically identified: Class I Regressed; Class II Early Maturation; Class III Mid Maturation; Class IV Late Maturation; Class V Regression. Monthly GSI, gV and Tse values were lower in January and February, the testis showed spermatozoa remains and a regenerating discontinuous germinal epithelium. In March spermiogenesis increased and proliferation of spermatogonia decreased. Male tropical gar followed a seasonal reproductive cycle, indicated by the monthly variation of the reproductive classes and the reproductive season processes observed, and for which temperature and rainfall seem to stimulate reproductive activity and spermiation.

Digestive system development and study of acid and alkaline protease digestive capacities using biochemical and molecular approaches in totoaba (Totoaba macdonaldi) larvae.

The present study aimed to describe and understand the development of the digestive system in totoaba (Totoaba macdonaldi) larvae from hatching to 40 days post-hatch (dph) from morphological and functional perspectives. At hatch, the digestive system of totoaba was undifferentiated. The anus and the mouth opened at 4 and 5 dph, respectively. During exogenous feeding, development of the esophagus, pancreas, liver and intestine was observed with a complete differentiation of all digestive organs. Expression and activity of trypsin and chymotrypsin were observed as early as at 1 dph, and increments in their expression and activity coincided with changes in food items (live and compound diets) and morpho-physiological development of the accessory digestive glands. In contrast, pepsin was detected later during development, which includes the appearance of the gastric glands between 24 and 28 dph. One peak in gene expression was detected at 16 dph, few days before the initial development of the stomach at 20 dph. A second peak of pepsin expression was detected at day 35, followed by a peak of activity at day 40, coinciding with the change from live to artificial food. Totoaba larvae showed a fully morphologically developed digestive system between 24 and 28 dph, as demonstrated by histological observations. However, gene expression and activity of alkaline and acid proteases were detected earlier, indicating the functionality of the exocrine pancreas and stomach before the complete morphological development of the digestive organs. These results showed that integrative studies are needed to fully understand the development of the digestive system from a morphological and functional point of views, since the histological organization of digestive structures does not reflect their real functionality. These results indicate that the digestive system of totoaba develops rapidly during the first days post-hatch, especially for alkaline proteases, and the stomach becomes functional between 20 and 24 dph allowing the weaning process to begin at this age.

[Microorganisms effect with probiotic potential in water quality and growth of the shrimp Litopenaeus vannamei (Decapoda: Penaeidae) in intensive culture]. / Efecto de microorganismos con potencial probiótico en la calidad del agua y el crecimiento de camarón Litopenaeus vannamei (Decapoda: Penaeidae) en cultivo intensivo.

The use of probiotics has gained acceptance in aquaculture, particularly in maintaining water quality and enhancing growth in organisms. This study analyzed the effect of the commercial (EM, Japan) natural product composed by (Rhodopseudomonas palustris, Lactobacillus plantarum, Lactobacillus casei and Saccharomyces cerevisiae) added to the water, in order to determine its effect in water quality, sediment and growth of L. vannamei under intensive culture. The evaluation included three treatments with a weekly addition of EM: i) tanks without probiotics (C), ii) tanks with a dose of 4 L/ha (EM1) and iii) tanks with a dose of 10 L/ha (EM2). The treatment C was carried out three times, while treatments EM1 and EM2 were carried out four times. A total of 4 350 shrimps were measured for total length and weight, to calculate total and porcentual weight gain, daily weight gain, specific growth rate (TCE), and food conversion factor (FCA); besides, the survival rate was estimated. The use of probiotics allowed a shorter harvest time in treatments EM1 (90 d) and EM2 (105 d) with relation to the treatment C (120d). Treatments EM1 and EM2 were within the recommended intervals for culture, with respect to treatment C. The use of probiotic bacteria significantly regulated pH (EM1, 8.03 +/- 0.33; EM2, 7.77 +/- 0.22; C, 9.08 +/- 0.35) and reduced nitrate concentration (EM1, 0.64 +/- 0.25 mg/L; EM2, 0.39 +/- 0.26 mg/L; C, 0.71 mg/L). Water pH mostly explained the variance with respect to the treatments. Treatment EM2 presented the greatest removal of organic matter (1.77 +/- 0.45%), whereas the contents of extractable phosphorus increased significantly in treatment EM1 with 21.6 +/- 7.99 mg/kg and in treatment EM2 with 21.6 +/- 8.45 mg/kg with control relation (14.3 +/- 5.47). The shrimp growth was influenced by dissolved oxygen, salinity and pH in the sediment, establishing that salinity was the most important variable in the weight with a negative association. Treatment EM1 recorded an improved TCE (2.69 +/- 0.35%/d) and FCA (1.46 +/- 0.20) with relation to the control treatment (TCE, 1.88 +/- 0.25%/d; FCA, 2.13 +/- 0.48). Survival was significantly greater in treatments containing probiotics with 61 +/- 8.76% and 60 +/- 10.5% for EM1 and EM2, respectively. This study indicated the positive effect obtained with the use of this commercial probiotic, to improve culture conditions and growth parameters in an intensive culture of L. vannamei.