Chymotrypsin Activity Research Articles

The Photoperiod Significantly Influences the Growth Rate, Digestive Efficiency, Immune Response, and Antioxidant Activities in the Juvenile Scalloped Spiny Lobster (Panulirus homarus)

This study aimed to elucidate the effects of different photoperiods (0 L:24 D, 6 L:18 D, 12 L:12 D, 18 L:6 D, 24 L:0 D, “Light (L) and Dark (D)”) on the growth performance and physiological responses of the juvenile scalloped spiny lobster (Panulirus homarus). Over a period of 56 days, parameters such as growth rate, digestive enzyme, immune enzyme, and antioxidant enzyme were meticulously evaluated in 90 lobsters subjected to these varying light conditions. The present study found no significant differences in survival rate (SR), molting frequency (MF), and meat yield production (MYP) among the various photoperiod treatments (p > 0.05). Notably, the highest weight gain rate (WGR) and specific growth rate (SGR) were observed under a 12 L:12 D photoperiod. In the continuous dark phase (0 L:24 D), pepsin (PEP) activity remained high in gastric tissues, while trypsin (TRYP) and chymotrypsin (CHT) activities reached the highest in hepatopancreas tissues. The α-amylase (AMS) activity in the hepatopancreas was most elevated under 18 L:6 D, and the optimal lipase (LPS) activity was recorded under 12 L:12 D. The activity of acid phosphatase (ACP) in the hepatopancreas was highest in the absence of light (0 L:24 D), whereas the activities of alkaline phosphatase (AKP) and lysozyme (LZM) were most effective under the 12 L:12 D photoperiod. The total antioxidant capacity (T-AOC), along with catalase (CAT) and superoxide dismutase (SOD) activities of the hepatopancreas reached the highest at 12 L:12 D. The highest activity of glutathione peroxidase (GSH-Px) was seen under 18 L:6 D. The concentration of malondialdehyde (MDA), a marker of oxidative stress, was found to be highest under 12 L:12 D. Consequently, this specific photoperiod is essential for achieving optimal growth and maintaining appropriate physiological balance in the scalloped spiny lobster during aquaculture. These findings provide a foundational guideline for establishing the lighting environment in the farming of the juvenile scalloped spiny lobster.

Feed form and perch design do not interact to production performance, gastrointestinal tract traits, behaviour and welfare of laying hens reared in enriched cages

ABSTRACT 1. It was hypothesised that perch material and design may affect utility and maintenance energy demand in laying hens, affecting their feed form preferences and daily feed consumption. Accordingly, perch design and feed form on hen performance, gastrointestinal tract functions and some behavioural and welfare-related traits were studied in laying hens (ATAK-S) reared in enriched colony cages from 24 to 40 weeks of age. 2. The experiment was a 2 × 2 factorial investigating two perch materials and design (circular steel or mushroom-shaped plastic) and feed form (mash or crumble). A total of 396 hens were randomly assigned to one of the four treatment groups with nine replicates each (11 birds per replicate). 3. Except for feeding behaviour and prevalence of foot pad dermatitis at 40 weeks of age, the modification of the perch design did not have a significant effect on the traits examined. Mushroom-shaped plastic perches reduced feeding behaviour (p < 0.01) and the incidence of foot pad dermatitis at 40 weeks of age (p < 0.001). 4. Performance traits were not affected by feed form. Intake, final body weight and FCR for crumble-fed laying hens were greater than those fed mash (p < 0.01). 5. Hens fed mash had higher (p < 0.01) relative gizzard weights along with lower (p < 0.05) pH values, pancreatic chymotrypsin, amylase and lipase activities (p < 0.05), and duodenal absorption surface areas (p < 0.01). Ultimately, this gave higher protein digestibility (p < 0.05) compared to those receiving crumble. 6. In conclusion, in enriched cage rearing systems, mashed feed was preferred over crumble to efficiently maintain productive performance. Compared to circular steel, plastic mushroom-shaped perches were associated with better footpad health and welfare.

Chymotrypsin and Trypsin Inhibitory Activity of Some Medicinal Plants Collected from Rize (Türkiye).

In this research, the evaluation of in vitro chymotrypsin and trypsin inhibitory activities of ten plant species collected from Rize were aimed, and fractions that showed strong activity were analyzed through HPLC. Daphne pontica L. and Mentha longifolia (L.) L. were found to have the highest chymotrypsin inhibitory activities (87.75 and 84.24 % inhibition). Similarly, the highest trypsin inhibitory activity was observed in D. pontica (%99.93 inhibition), followed by Sambucus ebulus L. flowers (87.47 % inhibition). Extracts showing strong enzyme inhibition were fractioned and subjected to activity tests. The highest chymotrypsin inhibitory activity was observed in the n-hexane fraction of D. pontica (%80.70 inhibition), while the highest trypsin inhibitory activity was found in the n-butanol fraction of S. ebulus (%86.81 inhibition). HPLC studies determined that the 80 % ethanol extract of D. pontica and its dichloromethane and ethyl acetate fractions contained umbelliferone. It was found that chlorogenic acid was present in the 80 % ethanol extracts of S. ebulus flowers. M. longifolia was found to contain chlorogenic acid, caffeic acid, luteolin-7-glucoside, and rosmarinic acid. M. longifolia has been identified as the plant exhibiting the highest antioxidant activity in ABTS and CUPRAC tests, consistent with its high phenolic and flavonoid content.

Rhodiola rosea L. improved intestinal digestive enzyme activities, inflammatory response, barrier and microbiota dysbiosis in Lateolabrax maculatus juveniles fed with high-carbohydrate diets

A 56-d feeding trial was conducted to evaluate the influences of Rhodiola rosea L. on digestive enzyme activities, intestinal barrier, inflammatory response, and microbiota dysbiosis in Lateolabrax maculatus juveniles (9.37 ± 0.03 g) fed with high-carbohydrate diets. Six diets were designed: a control diet (20% corn starch, Control), high-carbohydrate diet (30% corn starch, HC1), and four high-carbohydrate diets supplemented with Rhodiola rosea L. at 30, 60, 90 and 120 mg/kg (HC2, HC3, HC4 and HC5, respectively). Compared with the control group, the HC1 diet remarkably increased α-amylase, lipase, and chymotrypsin activities in the intestine (p < 0.05), as well as the mRNA levels of Claudin-15, NF-κB, TNF-α, IL-1β, and IL-8 (p < 0.05) and the relative abundance of Proteobacteria and Photobacterium in the intestine, which belong to the phylum and genus level, respectively. But the opposite trend was found in muscular thickness and villus lengths (p < 0.05), the mRNA levels of Occludin, ZO-1, and TGF-β (p < 0.05), at the level of phylum and genus level in the HC1 group, and the relative abundance of Firmicutes, Bacteroidetes, and Bacillus in the intestine compared with the control group. Intestinal chymotrypsin activity was significantly higher in the HC3 group and intestinal muscular thickness and villus lengths were also significantly higher in the HC2, HC3, HC4, and HC5 groups compared to the HC1 group (p < 0.05). In addition, Occludin mRNA expression in the intestine was significantly increased in the HC2, HC4, and HC5 groups compared to the HC1 group. ZO-1 and TGF-β mRNA expression in the intestine were significantly increased in the HC2, HC3, HC4, and HC5 groups compared to the HC1 group (p < 0.05). At the phylum level, the relative abundance of Firmicutes and Bacteroidetes was higher in the intestine in the HC2, HC3, HC4, and HC5 groups than that in the HC1 group. On the contrary, intestinal lipase and chymotrypsin activities were significantly decreased in the HC2 group compared to the HC1 group, respectively (p < 0.05). The Claudin-15, NF-κB, TNF-α, IL-1β, and IL-8 mRNA expression in the intestine were significantly decreased in the HC2, HC3, HC4, and HC5 groups compared to the HC1 group (p < 0.05). Besides, at the genus level, compared to the HC1 group, the relative abundance of Photobacterium in the intestine and the diversity of the intestinal microbiota in the HC2, HC3, HC4, and HC5 groups were all decreased. In conclusion, these results demonstrated that the addition of Rhodiola rosea L. in high-carbohydrate diets can improve intestinal digestive enzyme activities, inflammatory response and intestinal barrier-related gene expression, and microbiota dysbiosis in L. maculatus. The suitable supplemental level of Rhodiola rosea L. in high-carbohydrate diets of L. maculatus is 60 mg/kg.

Dietary Selenium Nanoparticles Improved Growth and Health Indices in Asian Seabass (Lates calcarifer) Juveniles Reared in High Saline Water.

A 60-day study was carried out to determine the effect of dietary selenium nanoparticles (SeNP) on growth, digestive enzymes, and health status of Asian seabass (Lates calcarifer, 46.5 ± 0.2 g) juveniles reared in high saline water (48 ppt). Five levels of SeNP were added to a basal diet (45% protein, 15% lipid), including control (0), 0.5 (SeNP0.5), 1.0 (SeNP1), 2 (SeNP2), and 4 (SeNP4) mg SeNP kg-1 diet. Fish were stocked into fifteen 2,000 L tanks (50 fish tank-1) filled with 1,800 L sand-filtered seawater (26.5 ± 1.5°C, 48.0 ± 0.2 ppt) in a flow-through system. Each dietary treatment was performed in three replicates. The growth rate positively increased in both linear and quadratic trends with increasing dietary SeNP level (P < 0.05). The liver Se concentration increased with increasing SeNP in diet (P < 0.05). Gut total protease, trypsin, chymotrypsin, alkaline phosphatase (ALP), lipase, and α-amylase activities were significantly enhanced in the SeNP4 group compared to the other treatments (P < 0.05). Antioxidant capacity improved in fish-fed SeNP2 and SeNP4 diets regarding catalase and superoxide dismutase activities and the liver glutathione content. Serum lysozyme and hemolytic activities and white blood cells' respiratory burst activity in the control were lower than in fish-fed SeNP-supplemented diets. Serum total protein, globulin, and globulin/albumin ratio in fish-fed SeNP1, SeNP2, and SeNP4 diets were higher than the other groups (P < 0.05). The interleukin-10 and granulocyte-macrophage colony-stimulating factor genes' relative transcription levels in the gut of fish-fed SeNP4 were higher than the other groups. Serum cholesterol, triglycerides, ALP, aspartate aminotransferase, alanine aminotransferase, and lactate dehydrogenase significantly decreased in fish with increasing SeNP content in the diet. In conclusion, supplementing the diet with 4 mg kg-1, SeNP was recommended to improve growth and health indices in L. calcarifer juveniles reared in high saline water (48 ppt).

Efficiency of protein combinations in diets for Rhamdia quelen: growth, digestive and metabolic biochemistry and nutrient deposition.

This study was conducted to determine the best combination of protein sources in diets for jundiá, based on growth, metabolism, and nutrient deposition. Five protein combinations were tested: casein + fish meal (control), casein + gelatin, casein + albumin, casein + albumin + fish meal, and albumin + fish meal, in diets containing 370 g Kg-1 of crude protein and 13.4 MJ Kg-1 of digestible energy. The fish (9.38 ± 0.12 g) were allocated in a water recirculation system at a density of 3.35 g L-1 per experimental unit and fed until apparent satiety for 40 days with the diets. The fish fed with the control diet had the highest final weight, specific growth rate, protein and feed efficiency ratio, protein retention, and best apparent feed conversion. On the other hand, fish that received casein + albumin and albumin + fish meal diets showed worse results in growth and body protein retention, low trypsin and chymotrypsin activity, and high intestinal amylase activity. Therefore, the combination referred to as control (casein + fish meal) conclusively provides the best rhythm for nutrient digestion and metabolism processes, enabling fish to reach greater growth and retention of body protein with low whole-fish fat content.

Cellulose acetate microwell plates for high-throughput colorimetric assays.

Single use plasticware (SUP) in scientific, diagnostic, and academic laboratories makes a significant contribution to plastic waste generation worldwide. Polystyrene (PS) microwell plates form a part of this waste. These plates are the backbone of high throughput colorimetric measurements in academic, research, and healthcare settings for detection/quantification of wide-ranging analytes including proteins, carbohydrates, nucleic acids, and enzyme activity. Polystyrene (PS) microwell plates serve as a platform for holding samples and reagents, where mixing initiates chemical reaction(s), and the ensuing color changes are quantified using a microplate reader. However, these plates are rarely reused or recycled, contributing to the staggering amounts of plastic waste generated in scientific laboratories. Here, we are reporting the fabrication of cellulose acetate (CA) microwell plates as a greener alternative to non-biodegradable PS plates and we demonstrate their application in colorimetric assays. These easy to fabricate, lighter weight, customizable, and environmentally friendly plates were fabricated in 96- and 384-well formats and made water impermeable through chemical treatment. The plates were tested in three different colorimetric analyses: (i) bicinchoninic acid assay (BCA) for protein quantification; (ii) chymotrypsin (CT) activity assay; and (iii) alkaline phosphatase (AP) activity assay. Color intensities were quantified using a freely available smartphone application, Spotxel® Reader (Sicasys Software GmbH). To benchmark the performance of this platform, the same assays were performed in commercial PS plates too and quantified using a UV/Vis microplate reader. The two systems yielded comparable linear correlation coefficients, LOD and LOQ values, thereby validating the CA plate-cell phone based analytical method. The CA microwell plates, coupled with smart phone optical data capture, provide greener, accessible, and scalable tools for all laboratory settings and are particularly well-suited for resource- and infrastructure-limited environments.

Chronic ammonia stress caused disorder of intestinal microbiota and damaged intestinal structure and function in yellow catfish (Pelteobagrus fulvidraco)

Ammonia is one of the major chemicals in aquatic environmental systems, and high concentrations of ammonia are toxic to fish. Also, ammonia stress can damage the intestine of fish. We wanted to know how chronic ammonia stress affects the intestinal microbiota and function of yellow catfish (Pelteobagrus fulvidraco). In this study, chronic ammonia stress (control group, 0.5 and 2.5 mg/L ammonia stress groups, continued 28 d) on intestinal structure, function and microbiota of the yellow catfish were explored. Chronic ammonia stress induced changes in the intestine microbiota according to 16 S high-throughput sequencing. The ACE, Shonnon and Chao indices of intestine microbiota showed an increasing trend in the 2.5 mg/L ammonia-stressed group. Ammonia stress also led to changes in the intestinal microbiota and the relative abundance of bacteria. We found the abundance of Flavobacteriaceae, Vibrio, Pseudomonas, Aeromonas and Streptococcus showed increasing tendency in ammonia stress groups. The H&E staining showed that chronic ammonia stress caused in marked epithelial cell necrosis and lamina propria edema and reduced width of intestinal villi significantly. We also found chronic ammonia stress significantly decreased in the number of goblet cells by PAS staining. In addition, ammonia stress caused significant decrease in activities of pepsin, chymotrypsin and trypsin. In summary, ammonia stress led to disruption of intestinal microbiota, causing damage of the intestinal structure and affecting the normal physiological function of the intestine.

Evaluation of pancreatic chymotrypsin activity for on-site prediction of clinically relevant postoperative pancreatic fistula

Although the risk of complications due to postoperative pancreatic fistula (POPF) have been evaluated based on the amylase level in drained ascitic fluid, this method has much room for improvement regarding diagnostic accuracy and facility of the measurement. This study aimed to investigate the clinical value of measuring pancreatic chymotrypsin activity for rapid and accurate prediction of POPF after pancreaticoduodenectomy. In 52 consecutive patients undergoing pancreaticoduodenectomy, the chymotrypsin activity in pancreatic juice was measured by calculating the increase in fluorescence intensity during the first 5min after activation with an enzyme-activatable fluorophore. The predictive value for clinically relevant POPF (CR-POPF) was compared between this technique and the conventional method based on the amylase level. According to receiver operating characteristic analyses, pancreatic chymotrypsin activity on postoperative day (POD) 3 measured with a multiplate reader had the highest predictive value for CR-POPF (area under the curve [AUC], 0.752; P<0.001), yielding 77.8% sensitivity and 68.8% specificity. The AUC and sensitivity/specificity of the amylase level in ascitic fluid on POD 3 were 0.695 (P=0.053) and 77.8%/41.2%, respectively. Multivariable analysis identified high pancreatic chymotrypsin activity on POD 3 as an independent risk factor for CR-POPF. Measurement of pancreatic chymotrypsin activity with a prototype portable fluorescence photometer could significantly predict CR-POPF (AUC, 0.731; P=0.010). Measurement of pancreatic chymotrypsin activity enabled accurate and rapid prediction of CR-POPF after pancreaticoduodenectomy. This can help surgeons to implement appropriate drain management at the patient's bedside without delay.

Fecal virus transplantation has more moderate effect than fecal microbiota transplantation on changing gut microbial structure in broiler chickens

Growing evidence of fecal microbiota transplantation (FMT) and fecal virus transplantation (FVT) provides a possibility to regulate animal health, whereas little is known about the impact of the 2 methods. This study aimed to investigate the effects of gut microbes on jejunal function in healthy broiler chickens, with the objective of establishing a theoretical basis for the application of FMT and FVT. Cecal feces from 28-day-old AA broilers were collected to prepare gavage juice for FMT and FVT. FMT for Group FM, FVT for group FV and PBS gavage for group CON, continuously treated for 6 days start at 5-day-old chicks. Samples were collected at d 11 and d 21. The results showed that the treatment d 2 and the overall fecal score in treatment groups were significantly lower than CON group (P < 0.05). The jejunum morphology showed that FMT increased crypt depth, decreased villus height, V/C (P < 0.05) and FVT increased villus height (P < 0.05) at d 11. At d 21, villus height and crypt depth significantly higher (P < 0.05) in group FM and group FV. The expression of Claudin1, Occludin, ZO2, and Muc2 in the FV group was significantly increased (P < 0.05) at 11-day-old. FMT increased the secretion of sIgA at 11-day-old, and this influence lasted up to 21-day-old (P < 0.05). At 11-day-old, the expression of b0+AT of basic amino acid transport carrier and chymotrypsin activity (P < 0.05) had a significant correlation. At 21 d of age, FVT significantly increased the expression of PepT1 and SGLT1 (P < 0.05). At 11-day-old, FM group showed significantly higher faith pd index (P = 0.004) and Shannon index (P = 0.037), and separated from FV and CON according to PCoA. Among differentiating bacteria, Bacteroides significantly enriched (P < 0.05) in group FM, which positively correlated with the expression of ZO2, Muc2, Occludin, and Claudin1; R_Ruminococcus, L_Ruminococcus, Butyricicoccuss significantly enriched (P < 0.05) in group CON, which significantly higher than processing groups, R_Ruminococcus and L_Ruminococcus negatively correlated with the expression of Occludin (P < 0.05), and R_Ruminococcus, Butyricicoccus negatively correlated with the expression of Claudin1 (P < 0.05). At 21-day-old, PCoA based on Bray-Curtis shows that microbes taxa of 3 groups are isolated with each other and treatment groups were significant different with CON group based on Unweighted UniFrac and weighted UniFrac. The expression of PepT1 was significantly negatively (P < 0.05) correlated with Ruminococcus, and the expression of sIgA was significantly negatively (P < 0.05) correlated with Parabacteroides. In conclusion, FMT regulated intestinal flora rapidly, while it had little effect on intestinal function and a higher potential damaging risk on jejunal. FVT regulated intestinal flora structure softer, improved tight junction expression, but the mechanism of action needs further exploration.

Effects of Dietary Sodium Propionate on Growth, Digestive Enzyme Activity, and Expression of Immune System Genes in Juveniles of Tropical Gar (Atractosteus tropicus)

We determined the effects of sodium propionate (SP) added to the diets of Atractosteus tropicus juveniles with respect to the growth, survival, digestive enzyme activity, and expression of genes that are associated with the immune system. Five treatments (0, 0.5, 1.0, 1.5, and 2.0%) were evaluated in triplicate on 180 fish (3.65 ± 0.12 g) distributed among 15 (70 L) tanks. The juveniles were fed five times a day with 5% feed in relation to the biomass of the organism. The treatment with 0.5% SP showed a final weight value of 25.7 ± 4.5 g, absolute weight of gain (AWG) of 21.93 ± 4.39 g, and specific growth rate (SGR) of 3.1 ± 0.26. Treatments with 1.5 and 2.0% SP showed the highest survival (91.6%). The control group (0%) showed a greater activity of lipases. There was a tendency that the highest activity of alkaline proteases and chymotrypsin occurred in the 0 and 0.5% treatments. The maximum relative expression of the genes ocln, muc2, and nod2 occurred in the 1.5% treatment. The inclusion of SP in the diet of A. tropicus juveniles could benefit the activity of some digestive enzymes as well as the expression of genes related to the function of the intestinal barrier, therefore benefitting the survival of the organisms.

Use of black soldier fly (Hermetia illucens) larvae meal in diets for gilthead seabream juveniles: Effects on growth-related gene expression, intermediary metabolism, digestive enzymes, and gut microbiota modulation

A study was conducted to evaluate the effects of dietary inclusion of defatted Hermetia illucens larvae meal (HM) on the expression of growth-related genes, plasma metabolites, activity of intermediary metabolism enzymes, and gut microbiota modulation in gilthead seabream (Sparus aurata) juveniles. Four experimental diets were formulated (43% crude protein, 18% crude lipids) with increasing levels of defatted HM: 0 (HM0), 15 (HM15), 30 (HM30), and 45% (HM45) replacing fishmeal (included at 34% in the control diet) at 0, 22, 60, and 100%, respectively. Fish were fed these diets for 67 days. At the end of the trial, the expression of muscle growth hormone receptors (ghr1 and ghr2), insulin-like growth factors (igf1 and igf2), cathepsins (cts-a and cts-b), and myogenic determination factor 1 (myod1) were not affected, while a downregulation of myogenic determination factor 2 (myod2) and upregulation of myostatin (mstn) was observed with HM inclusion. Plasma metabolites profile was unaffected by diet composition, except for a linear decrease trend in total lipids. A linear increase of liver alanine aminotransferase (ALT) and glutamate dehydrogenase (GDH) activity and a linear decrease of glucose 6-phosphate dehydrogenase (G6PDH), malic enzyme (ME), and (FAS) activity were observed with HM inclusion. The activity of β-hydroxy acyl-CoA dehydrogenase (HOAD) followed a quadratic trend, being higher in fish fed with diet HM15 than in the control. Likewise, amylase, trypsin, and chymotrypsin activity were higher in the posterior intestine of fish fed with diet HM15. Digesta of fish fed the HM-containing diets showed an increase in the number of operational taxonomic units, microbial richness, and diversity. Conversely, a reduction in these indexes was observed in the mucosa. Among the identified bacteria, Firmicutes represented the predominant phyla. The inclusion of HM increased the abundance of Oceanobacillus, Lederbergia, Bacillus, and Corynebacterium in the digesta while Lactobacillus and Caldalkalibacillus abundance was reduced. Overall, total FM replacement with defatted HM had no major effects on plasma metabolites and muscle growth-related genes but induced a few differences in intermediary metabolism and digestive enzymes activity. HM inclusion improved gut digesta microbiota richness and diversity, promoting the growth of potential beneficial bacteria.

Mechanisms of Digestive Enzyme Response to Acute Salinity Stress in Juvenile Yellowfin Tuna (Thunnus albacares).

This study investigates the effect of a sudden change in salinity for 48 h on the digestive enzyme activity of juvenile yellowfin tuna. The treatment included a control salinity of 32‱ in natural seawater and an experimental salinity of 29‱. Acute stress experiments were carried out on 72 juvenile yellowfin tuna (646.52 ± 66.32 g) for 48 h to determine changes in digestive enzyme activity in different intestinal sections over time (0 h, 12 h, 24 h, 48 h). The activities of pepsin, trypsin, α-amylase, lipase, and chymotrypsin in the digestive organs (stomach, foregut, and pyloric ceca) of juvenile yellowfin tuna were measured. Pepsin and pancreatic protease in the experimental group were significantly lower than in the control group (p < 0.05). α-amylase showed a fluctuating trend of decreasing and then increasing, and its activity trend was pyloric ceca > foregut > stomach. The lipase activity of gastric tissues decreased at the beginning and then increased, reaching a minimum at 24 h (2.74 ± 1.99 U·g protein-1). The change of lipase in the pyloric ceca and foregut was increasing and then decreasing. The lipase activity trend was pyloric ceca > foregut > stomach. The chymotrypsin showed a decreasing and increasing trend and then stabilized at 48 h with a pattern of pyloric ceca > foregut > stomach. Similarly, the gut villi morphology was not significantly altered in the acutely salinity-stressed compared to the non-salinity-stressed. This study suggests that salinity may change the digestive function of juvenile yellowfin tuna, thereby affecting fish feeding, growth, and development. On the contrary, yellowfin tuna is highly adapted to 29‱ salinity. However, excessive stress may negatively affect digestive enzyme activity and reduce fish digestibility. This study may provide a scientific basis for a coastal aquaculture water environment for yellowfin tuna farming, which may guide the development and cultivation of aquaculture.

Raw Fish-based Moist Pellet and Extruded Pellet on Growth, Levels and Expression of GH/IGF Axis, Intestinal Enzyme Activity and Gastrointestinal Morphology of Olive Flounder (Paralichthys olivaceus): An Actual Field Study

A six-month feeding trial was conducted to investigate the effect of moist pellet (MP) and commercial extruded pellet (EP) on the somatic growth, level and expression of GH/IGF axis, intestinal enzyme activity and gastrointestinal morphology of olive flounder (Paralichthys olivaceus) reared under actual farm conditions. MP was a raw fish-based diet composed of mackerel and cutlass fish while EP diet was a locally-produced commercial feed. Weight growth rate showed no significant difference (P&gt;0.05), although EP-fed fish had a better somatic yield. Feed conversion ratio was better and condition factor has significantly improved when given EP diet (P&lt;0.05). Plasma level of growth hormone and insulin-like growth factor 1 was comparably similar between diets, but EP-fed olive flounder had a higher hepatic IGF-1 expression (P&lt;0.05). The activity of trypsin, chymotrypsin and lipase were insignificantly influenced by diets (P&gt;0.05). The overall gastrointestinal morphology was in similitude regardless of diets, but EP- fed fish had a longer intestinal villi length and goblet cell count in the pyloric caeca (P&lt;0.05). These dietary induced differences are discussed to have partly promoted better growth in EP diet. The findings suggest an equivalent, or better utilization of EP diet in the actual farm operation.

Effect of dietary supplementation of the quenching enzyme AiiO-AIO6 on growth performance, digestion and absorption capacity, and intestinal barrier function of grass carp (Ctenopharyngodon idella)

Quenching enzymes have attracted wide attention as one of the alternatives to antibiotics because their positive effects on inhibiting pathogenic microorganism. Therefore, the objective of this study was to investigate the impact of dietary quenching enzyme AiiO-AIO6 on the growth performance, digestion and absorption capacity, intestinal structural integrity, and gut microbiota composition in juvenile grass carp. Healthy grass carp (4.73 ± 0.01 g) were assigned to 36 fish tanks (1.2 × 0.6 × 0.9 m), randomly divided into 6 groups, and fed on six diets with different levels of AiiO-AIO6 (0, 2.5, 5.0, 7.5, 10.0 and 12.5 U/g feed) for a duration of 10 weeks. The present findings demonstrated that dietary AiiO-AIO6 supplementation positively influenced the final body weight (FBW), specific growth rate (SGR), percent weight gain (PWG), and feed intake (FI) of juvenile grass carp. Moreover, our results indicated that the appropriate dietary AiiO-AIO6 supplementation promoted the number of goblet cells and improved the activities of chymotrypsin, trypsin, lipase, amylase, Na+-K+-ATPase, γ-GT and CK, which ultimately contributed to the enhancement of digestion and absorption capacity of juvenile grass carp. Additionally, the present study further found that dietary AiiO-AIO6 supplementation up-regulated the relative mRNA levels of tight junctions (TJs) and adhesive junctions (AJs) proteins, including ZO-1, occludin, claudin-b, −c, −f, −7a, −7b, −11, −12, α-catenin, nectin, afadin, and E-cadherin, thereby enhancing the physical barrier function, potentially attributed to the reduction in abundance of intestinal pathogenic bacteria, such as Pseudomonas. Ultimately, the optimal level of AiiO-AIO6 for juvenile grass carp, as determined by the percent weight gain (PWG), was found to be 4.5 U/g.

Effects of dietary thiamin (vitamin B1) on the growth performance, serum biochemical factors, immune response, and antioxidant activity of great sturgeon (Huso huso) juveniles.

The present research evaluated the positive effects of dietary thiamin (vitamin B1) levels on the growth performance, serum biochemistry factors, immune response, and antioxidant activity of great sturgeon (Huso huso) juveniles. Thiamin was included in diets with levels of 0 (control, T0), 7 (T7), 15 (T15), and 25 (T25) mg/kg diet. Measurements of thiamin levels in diets indicated that they contained 1.80 (T0), 8.02 (T7), 16.2 (T15), and 26.6 (T25) mg thiamin/kg feed. Sturgeon juveniles (240 individuals) with average weight of 44.8 ± 1.96g were distributed into 12 tanks, and fed with the experimental diets for 8 weeks. Final weight, body weight gain (%), specific growth rate, and feed conversion ratio (FCR) of great sturgeon were significantly influenced by dietary thiamin levels, and the maximum fish performance (P < 0.05) was obtained at a level of 15mg/kg diet. The trypsin, chymotrypsin, creatine kinase, lipase, α-amylase, and alkaline phosphatase activities were notably (P < 0.05) affected by the dietary thiamin levels. The glucose content was not significantly (P > 0.05) different among the experimental treatments. Diets supplemented with thiamine increased significantly (P < 0.05) triglyceride, cholesterol, and total protein levels accompanied with significant (P < 0.05) decreases in aminotransferase aspartate and alanine aminotransferase activities. Serum antioxidant enzymes were remarkably (P < 0.05) higher, while serum malondialdehyde was significantly (P < 0.05) lower in the thiamin-treated fish compared with the control group. Total immunoglobulin, lysozyme, and ACH50 values were significantly (P < 0.05) higher in fish fed with thiamin-supplemented diets than in the control group. The results of the present study demonstrated that dietary thiamin have an important role in enhancing the growth performance, immune response, and antioxidant activity of great sturgeon. Based on the regression fitting curve of final weight, weight gain, specific growth rate, and FCR values, the optimal level of thiamin is found to be 15.0-17.5mg/kg diet.

Expression patterns and antifungal function study of KaSPI in Mythimna separata.

Kazal-type serine protease inhibitors (KaSPI) play important roles in insect growth, development, digestion, metabolism and immune defence. In this study, based on the transcriptome of Mythimna separata, the cDNA sequence of MsKaSPI with Kazal domain was uploaded to GenBank (MN931651). Spatial and temporal expression analysis showed that MsKaSPI was expressed at different developmental stages and different tissues, and it was induced by 20-hydroxyecdysone in third-instar larvae of M. separata. After 24 h infection by Beauveria bassiana, the expression level of MsKaSPI and the corresponding MsKaSPI content were significantly up-regulated, being 6.42-fold and 1.91-fold to the control group, respectively, while the activities of serine protease, trypsin and chymotrypsin were inhibited. After RNA interference interfered with MsKaSPI for 6 h, the expression decreased by 73.44%, the corresponding content of MsKaSPI protein decreased by 55.66% after 12 h, and the activities of serine protease and trypsin were significantly enhanced. Meanwhile, both the larval and pupal stages of M. separata were prolonged, the weights were reduced and the number of eggs per female decreased by 181. Beauveria bassiana infection also increased the mortality of MsKaSPI-silenced M. separata by 18.96%. These prove MsKaSPI can not only result in slow growth and low fecundity of M. separata by regulating the activity of related protease, but also participate in the resistance to pathogenic fungi by regulating the serine protease inhibitor content and the activities of related serine protease.

A constitutive serine protease inhibitor suppresses herbivore performance in tea (Camellia sinensis).

Protease inhibitors promote herbivore resistance in diverse plant species. Although many inducible protease inhibitors have been identified, there are limited reports available on the biological relevance and molecular basis of constitutive protease inhibitors in herbivore resistance. Here, we identified a serine protease inhibitor, CsSERPIN1, from the tea plant (Camellia sinensis). Expression of CsSERPIN1 was not strongly affected by the assessed biotic and abiotic stresses. In vitro and in vivo experiments showed that CsSERPIN1 strongly inhibited the activities of digestive protease activities of trypsin and chymotrypsin. Transient or heterologous expression of CsSERPIN1 significantly reduced herbivory by two destructive herbivores, the tea geometrid and fall armyworm, in tea and Arabidopsis plants, respectively. The expression of CsSERPIN1 in Arabidopsis did not negatively influence the growth of the plants under the measured parameters. Our findings suggest that CsSERPIN1 can inactivate gut digestive proteases and suppress the growth and development of herbivores, making it a promising candidate for pest prevention in agriculture.

Partial characterization of digestive proteases in the river prawn Macrobrachium americanum () (Decapoda: Caridea: Palaemonidae) in Mexico

Abstract We characterized the digestive enzymes in adults of Macrobrachium americanum (Spence Bate, 1868) in southeastern Mexico. The digestive enzyme extracts were made from the specimens’ hepatopancreas. Alkaline proteases, trypsin, chymotrypsin, carboxypeptidase A, leucine aminopeptidase, lipases, α-amylase and alkaline phosphatase activities were determined, as well as the percentage of inhibition, pH, and temperature stabilities through biochemical and electrophoretic techniques. The maximum digestive activity of proteases was at 35 °C and pH 11. Alkaline digestive proteases were highly stable at pHs 4, 9 and 11 at 55 °C after 30 min pre-incubation. PMSF inhibited two bands with proteolytic activity (35.1 and 23.2 kDa), and SBT1 inhibited all bands, including the one with the highest molecular weight (64.2 kDa). We concluded that enzymes in M. americanum coincide with those of omnivorous decapods with a tendency to carnivory.

Comparative study on the valorization of Sargassum from the Mexican Caribbean coast and Gulf of California as an ingredient on healthy diets for shrimp farming

Sargassum biomass is a potential ingredient for aquaculture formulations due to its high nutritional value and availability, as it contains a variety of essential nutrients including proteins, carbohydrates, lipids, vitamins, antioxidants, fiber, and minerals, which are essential for aquatic growing, development and health parameters including digestibility and immune response against diseases. Therefore, in this work, five experimental diets were formulated considering 2.5 % and 5 % levels of inclusion of Sargassum spp. from the Mexican Caribbean coasts (SC), and from the Gulf of California (SG), and one reference diet as a control. The characterization of the raw material and the proximal analysis of the five diets were evaluated according to standard reference methods. A 35-day feeding trial was conducted using juvenile Litopenaeus vannamei showed a shrimp survival rate of 97 ± 6 % (p > 0.05) on SC. Excellent weight gain (WG) performance of SC and SG diets at 2.5 % of inclusion resulted in >10 % WG compared to the control. The specific activity of amylase, lipase, trypsin, and chymotrypsin showed a significant increase by including Sargassum in diets, compared with the reference diet. Incorporating Sargassum at 2.5 % in feed formulations for shrimps showed a great opportunity to use it as an ingredient and improved production yields. The digestion and absorption parameters were represented by the enzymatic activity in which the diets based on Sargassum spp. from the Mexican Caribbean coast (SC) have resulted in the best performance.