Enzymatic Digestion Research Articles

Consequences of dietary red pepper and radish oils’ supplementation on growth, carcass traits, blood indices, digestive enzymes activity and intestinal microbial load of broilers

Consequences of dietary red pepper and radish oils’ supplementation on growth, carcass traits, blood indices, digestive enzymes activity and intestinal microbial load of broilers

Umami-enhancing effect of Agaricus bisporus-pork bone stocks based on the EUC value and sensory evaluation

In this study, Agaricus bisporus and pork bone stocks (PBS) were enzymatically digested, and the equivalent umami concentration (EUC) value, sensory evaluation and hydrolysis degree (DH) were used as indexes to find the optimal enzyme digestion conditions. Finally, the optimal enzymatic solution was mixed and analyzed for flavor compounds in the optimal enzymatic solution before and after Maillard reaction (MR) by GC × GC-O-MS. The results showed that the enzymatic digestion of mushroom stocks (MS) with a combination of Cellulase-Protamex™ (C-P) yielded high levels of flavor amino acids (40.733 ± 0.340 g/L), and 5′-nucleotides (12.807 ± 0.239 mg/g), and EUC values up to 1755.786 ± 40.759 g/100 g. Meanwhile, the simultaneous enzymatic hydrolysis of PBS with a combination of Flavourzyme™-Protamex™ (F-P) yielded high levels of flavor amino acids (51.265 ± 0.550 g/L) and 5′-nucleotides (17.477 ± 0.260 mg/g) with EUC values up to 3420.619 ± 114.193 g/100 g. Using the DH and sensory evaluation as indicators, the results showed that the greatest DH and the highest sensory scores were obtained with the addition of C-P-MS and F-P-MS at 7:3 (m/m). Finally, the compounds with large changes in concentration were mainly mushroom flavored 1-octen-3-ol (2208.614 ± 12.879 μg/kg), sweet-flavored 3-octanone (269.871 ± 11.691 μg/kg), floral-flavored benzyl alcohol (156.704 ± 14.859 μg/kg), green flavored 2-pentylfuran (190.470 ± 12.934 μg/kg), and almond-flavored furfural (523.555 ± 5.674 μg/kg).

Cloning, Expression, and Purification of Phage Display-Selected Fab for Biophysical and Biological Studies.

The antigen-binding fragment (Fab) is the ∼50-kDa monovalent arm of an antibody molecule. In the laboratory, the Fab can be produced via either enzymatic digestion or recombinant expression, and its use facilitates the accurate assessment of affinity and specificity of monoclonal antibodies. The high melting temperature of the Fab, together with its low tendency to aggregate and ready conversion to natural and nonnatural immunoglobulin (Ig) formats (without affecting antigen binding properties), have made it a preferred format for phage display, as well as a tool for accurate assessment of affinity, specificity, and developability of monoclonal antibodies. Here, we outline a strategy to clone, express, and purify human or chimeric nonhuman/human Fabs that have previously been selected by phage display. Fabs purified using this approach, which results in milligram amounts, enable a variety of downstream biophysical and biological assays that ultimately inform the success of phage display library generation and selection.

Unveiling the Potential Bioactive Peptides Derived From Goat Casein Hydrolysates Based on In Silico Analyses

ABSTRACTBioactive peptides (BAPs) have attracted considerable interest in scientific research due to their heterogeneity in sequence and structure, which underpins various biological functionalities. In this context, goat casein, an abundant by‐product of the dairy industry, emerges as a valuable source of BAPs. The present study undertook a meticulous evaluation of the bioactive potentials of goat casein‐derived peptides through an integrated approach combining computational simulations, high‐throughput screening, and molecular docking techniques. The initial phase involved the enzymatic digestion of goat milk casein using trypsin, followed by the identification of peptides via liquid chromatography–tandem mass spectrometry (LC–MS/MS), uncovering a total of 597 peptides. Subsequent prioritization using the PeptideRanker algorithm identified 70 peptides exhibiting potential bioactivity, as denoted by scores above 0.8. Advanced screening employing the BIOPEP database and the AutoDock and CAMPR4 tools facilitated the elucidation of 16 antioxidant, 59 hypotensive, 63 hypoglycemic, 70 hypolipidemic, and 25 antimicrobial peptides. Molecular docking studies further elucidated the spontaneous nature of the interactions between the peptides and their respective receptors, predominantly mediated by hydrogen bonding and hydrophobic interactions. Four peptides specifying all activities simultaneously were synthesized, and their activities were verified by in vitro experiments. These results not only highlight an effective strategy for the high‐throughput screening of goat casein‐derived peptides but also underscore the potential of utilizing casein as a viable source of functional food ingredients. This study thereby contributes significantly to the expanding field of functional food research, suggesting a sustainable approach to explore the potential of dairy by‐products.

Probing the role of ligation and exonuclease digestion towards non-specific amplification in bioanalytical RCA assays.

Non-specific amplification (NSA, amplification in the absence of a target analyte) in bioanalytical rolling circle amplification (RCA) assays, especially those involving pre-synthesized circular DNA (cDNA), affects its analytical sensitivity. Despite extensive development of RCA-based bioanalytical methods, the NSA in RCA remains uncharacterized in terms of its magnitude or origin. NSA may originate from inefficient ligation or succeeding cDNA purification steps. This study comprehensively quantifies NSA across several ligation and digestion techniques for the first time since the innovation of RCA. To quantify the NSA in RCA, cDNAs were prepared using self-annealing, splint-padlock, or cohesive end ligations. The cDNAs were then subjected to nine different exonuclease digestion steps and quantified for NSA under linear as well as hyperbranched RCA conditions. We investigated buffer compositions, divalent ion concentrations, single or dual enzyme digestion, cohesive end lengths, and splint lengths. The optimized conditions successfully mitigated absolute NSA by 30-100-fold and relative NSA (normalized against primer-assisted RCA) to ∼5%. Besides understanding the mechanistic origin of NSA, novel aspects of enzyme-substrate selectivity, buffer composition, and the role of divalent ions were discovered. With increasing bioanalytical RCA applications, this study will help standardize NSA-free assays.

Production of Sugars and Ethanol from Acid–Alkaline-Pretreated Agave sisalana Residue

Drylands in Brazil have been exploring sisal (Agave sisalana) as an essential source of income. However, the solid residues generated because of this activity still need suitable destinations; therefore, research has been carried out to transform them into added-value products. Therefore, the present study evaluated the potential of sisal or agave solid residue as a precursor feedstock for second-generation ethanol production. Acid and acid–alkaline pretreatments were carried out on sisal residues to enrich the biomass with cellulose and maximize enzymatic digestibility. Second-generation ethanol production was carried out using Semi-simultaneous saccharification and fermentation (SSSF). Regardless of catalyst dosage and incubation time, oxalic acid pretreatments generated samples with a similar chemical composition to those pretreated with sulfuric acid. However, samples pretreated with oxalic acid showed lower enzymatic digestibility. Samples pretreated with oxalic acid and sodium hydroxide obtained 14.28 g/L of glucose and cellulose conversion of 79.1% (at 5% solids), while 21.49 g/L glucose and 91.2% of cellulose conversion were obtained in the hydrolysis of pretreated samples with sulfuric acid and sodium hydroxide combined pretreatments. The pretreatment sequence efficiently reduced cellulase dosage from 20 to 10 FPU/g without compromising sugar release. SSSF achieved maximum production of 40 g/L ethanol and 43% ethanol conversion using 30% solids and gradually adding biomass and cellulases.

Effect of Dietary Supplementation of Glycerol Monolaurate on Growth Performance, Digestive Enzymes, Serum Immune and Antioxidant Parameters, and Intestinal Morphology in Black Sea Bream.

An eight-week feeding trial was conducted to examine the impact of dietary supplementation with glycerol monolaurate (GML) on juvenile black sea bream. A basal diet was formulated containing 24% fish meal, while five additional diets were prepared, each supplemented with varying levels of GML: GML1 (0.01%), GML2 (0.02%), GML3 (0.04%), GML4 (0.08%), and GML5 (0.16%). Triplicate tanks were randomly allocated to each diet, each containing 20 fish with an initial weight of 1.55 ± 0.05 g. By the trial's end, the GML3 group displayed a notably higher final body weight (FBW), weight gain (WG), specific growth rate (SGR), and protein efficiency ratio (PER) compared to the other groups (p < 0.05), but the FCR was significantly higher in the control group. However, no significant differences were observed in the MFI, PPV, CF, HSI, IPF, VSI, or SR among the groups (p > 0.05). Regarding the proximate compositions of the dorsal muscle and whole body, no substantial differences were observed across the groups (p > 0.05). Additionally, there were no significant variations in digestive enzyme activity (p > 0.05), serum immune, or biochemical parameters in the midgut and hindgut among the treatment groups. But in the serum immune response IgM, C3 and C4 were significantly higher in the GML3 group as compared to the other groups (p < 0.05). However, the GML3 group exhibited significantly greater fore-intestinal villus height, crypt depth, villus height per crypt depth, and the number of goblet cells per villus compared to the other groups (p < 0.05). Overall, GML supplementation, particularly GML3, significantly improved growth indicators like the final body weight and intestinal morphology. While certain parameters remained unaffected, these findings suggest GML's potential as a beneficial dietary supplement in fish diets.

Functional characterization of chalcone reductase gene CHR3 through RNAi

In this experiment, the soybean genome "Jilin 30" was used as a template to clone the target gene CHR3 using specific primers. The sense fragment, antisense fragment, and intron of the target gene were ligated into pCAMBIA3300 through double enzyme digestion to form a stem loop structure. We then transferred the constructed RNAi vector pCAMBIA3301-CHR3 RNAi into the recipient soybean genome via Agrobacterium mediated method, specifically reducing the expression of the target gene CHR3. By genetic transformation, positive plants were obtained, and molecular biology methods such as Southern blotting, real-time quantitative PCR, isoliquiritigenin analysis, and identification of resistance to Phytophthora root rot were used to detect the expression level of the CHR3 gene in the positive plants and study its function.The RNAi expression vector pCAMBIA3300-CHR3-RNAi was successfully constructed using the sense and antisense fragments of 428 bp CHR3 gene and an intron of 110 bp was inserted to form the stem-loop structure of the RNAi vector. PCR was used to detect 6 lines of the T1 generation and 15 lines of the T2 generation. The results of Southern analysis confirmed the integration of CHR3 and marker genes into the soybean genome. The expression CHR3 gene in the T1 genotypes decreased by 13.1 to 58.3%; the expression in the root decreased by 0.9 to 47.4%; the expression in the leaf decreased by 20 to 44.2%; and the expression in the stem decreased by 7.6 to 23%. The content of isoliquiritigenin decreased by 26.7% as transgenic shoots had a lower content of isoliquiritigenin and reduced resistance to Phytophthora sojae. Bangladesh J. Bot. 53(3): 627-633, 2024 (September) Special

Sea cucumbers and their symbiotic microbiome have evolved to feed on seabed sediments

Sea cucumbers are predominant deposit feeders in benthic ecosystems, providing protective benefits to coral reefs by reducing disease prevalence. However, how they receive sufficient nutrition from seabed sediments remains poorly understood. Here, we investigate Holothuria leucospilota, an ecologically significant tropical sea cucumber, to elucidate digestive mechanisms underlying marine deposit-feeding. Genomic analysis reveals intriguing evolutionary adaptation characterized by an expansion of digestive carbohydrase genes and a contraction of digestive protease genes, suggesting specialization in digesting microalgae. Developmentally, two pivotal dietary shifts, namely, from endogenous nutrition to planktonic feeding, and from planktonic feeding to deposit feeding, induce changes in digestive tract enzyme profiles, with adults mainly expressing carbohydrases and lipases. A nuanced symbiotic relationship exists between gut microbiota and the host, namely, specific resident bacteria supply crucial enzymes for food digestion, while other bacteria are digested and provide assimilable nutrients. Our study further identifies Holothuroidea lineage-specific lysozymes that are restrictedly expressed in the intestines to support bacterial digestion. Overall, this work advances our knowledge of the evolutionary innovations in the sea cucumber digestive system which enable them to efficiently utilize nutrients from seabed sediments and promote food recycling within marine ecosystems.

Extraction, pharmacological prediction and experimental verification of Coreopsis tinctoria flowers

Coreopsis tinctoria (C. tinctoria) is a traditional plant with high medicinal value and edible value. It has the effect of invigorating spleen and stomach and has the potential to develop into flower tea. Therefore, the extraction process of C. tinctoria extract (CTA) was optimised by analytic hierarchy process (AHP) combined with orthogonal test, and its activity was predicted by network pharmacology and verified by experiments. The optimal extraction conditions: temperature of 80 °C, extraction period of 10 min, solid-liquid ratio of 1:70 and times 2. CTA had no gastrointestinal irritation and can significantly increase the faecal water content of mice, inhibit amylase activity, increase protease, xylanase and cellulase activity, increase the number of intestinal Lactobacillus and Bifidobacterium, reduce the number of Escherichia coli and Enterococcus. In summary, CTA can effectively improve the intestinal digestive enzyme activity and the number of probiotics and has the potential to treat gastrointestinal diseases.

GingisREX: A Complementary Enzyme for the Detection of Bacterial Proteins.

Reliable enzymatic digestion underscores successful mass-spectrometry-based proteomics experiments. In this study, we compare the use of the arginine-specific protease, GingisREX, against a more traditional approach in the identification of Escherichia coli proteins. An increased number of protein identifications were noted when GingisREX was used compared to a trypsin/lys-C mixture. This improvement was attributed to the generation of fewer peptides per protein, resulting in a simpler peptide mixture. Furthermore, GingisREX exhibited increased digestion efficiency, fewer missed cleavages, and improved MS/MS data quality for higher molecular weight peptides. The data here establish GingisREX to be a protease complementary to trypsin for enhanced detection of bacterial proteins. With further optimization, GingisREX could prove to be an effective alternative to trypsin for identifying host cell proteins in biotherapeutics.

Effects of Alanyl-Glutamine Dipeptide Supplementation on Growth Performance, Nutrient Digestibility, Digestive Enzyme Activity, Immunity, and Antioxidant Status in Growing Laying Hens.

Alanyl-glutamine (Aln-Gln), a highly soluble and stable Glutamine-dipeptide, is known to improve the performance of poultry birds. This study aimed to investigate the effect of Aln-Gln during the rearing period on growth performance, nutrient digestibility, digestive enzyme activity, immunity, antioxidant status and relative gene expression of Hy-Line brown hens. A total of 480 healthy day-old Hy-line brown chicks with similar body weights were randomly divided into four dietary groups (8 replicates/group and 15 birds/replicate). Groups A, B, C and D were fed diets containing 0%, 0.1%, 0.2% and 0.3% Aln-Gln, respectively, for 6 weeks. The body weight (BW) and average daily gain (ADG) were higher in hens fed test diets compared with the control (p < 0.05). The feed conversion ratio (FCR) was better in test groups as compared to the control group (p < 0.05). The ADFI showed no significant difference between the groups. Dietary treatments had no effect on dry matter (DM), organic matter (OM) and crude fiber (CF) digestibility. The Aln-Gln also improved gross energy (GE) and crude protein (CP) digestibility (p < 0.05). It has also increased IgG levels in groups C and D. IgM levels were similar to the control in B, C and D. The Aln-Gln increased IL-1 in B and C, IL-2 in C and D, and IL-6 in all test groups (p < 0.05). The supplementation of Aln-Gln had no effect on serum antioxidant indices like CAT, MDA, GSH-PX, GSH, and SOD in 42-day-old growing hens. Aln-Gln supplementation had no significant effect (p > 0.05) on the activity of amylase and lipase, however, a significant improvement (p < 0.05) in the activities of trypsin and chymotrypsin was observed in the test groups. Supplemented Aln-Gln levels in the birds' diets led to an increase in the expression of genes related to growth factors (IGF-1, IGFBP-5), immune markers (IL-1, IL-2, IL-6) and antioxidant status (GSH-Px1), as compared to control group. Aln-Gln supplementation in Hy-Line brown hens during their growing period improved growth, nutrient digestibility, immunity and digestive enzymes activity. These findings suggest that Aln-Gln is a promising dietary additive for enhancing poultry performance.

Association between empty nose syndrome and laryngopharyngeal reflux disease: a preliminary cohort study.

To investigate the association between laryngopharyngeal reflux disease (LPRD) and Empty Nose Syndrome (ENS). Nasal and laryngopharyngeal reflux symptoms were investigated in patients with ENS. Symptoms were evaluated with reflux symptom score-12 (RSS-12), nasal obstruction symptom evaluation (NOSE), empty nose syndrome 6-item questionnaire (ENS6Q), empty nose syndrome index (ENSI), and sinonasal outcome tool-22 (SNOT-22). The anxiety and depression were assessed with the general anxiety disorder-7 (GAD-7), and patient health questionnaire-9 (PHQ-9). A study of association was conducted between demographics and patient-reported outcome questionnaires. Forty-one ENS patients were included (20 females (48.8%)). The control groups included 27 patients with rhinitis/rhinosinusitis and 36 asymptomatic individuals. The ENSI and ENS6Q detected ENS in 97.6% and 90.2% of cases, respectively. The mean scores of ENSI, ENS6Q, RSS-12, NOSE, and SNOT-22 were significantly higher in the ENS group compared to controls. The prevalence of suspected LPRD was 90.2% in the ENS group, which was significantly higher compared to controls. The prevalence of mild, moderate, moderately severe, and severe depression in ENS patients was 7.3% (n = 3), 4.9% (n = 2), 39.0% (n = 16), and 46.3% (n = 19), respectively. RSS-12 reported significant and high associations with the ENS6Q (rs=0.939; p = .001) and ENSI (rs=0.699; p = .001). LPRD symptoms and prevalence were significantly higher in ENS patients compared to controls. Future controlled studies are needed to investigate the prevalence of LPRD in ENS patients through objective approaches (impedance-pH monitoring, nasal digestive enzyme measurements).

Effect of Complexation with Different Molecular Weights of Oat β-Glucan and Sea Buckthorn Flavonoid on the Digestion of Rice Flour.

The complex of oat β-glucan (OBG) and flavonoids hampered the digestion of starch-based food and retarded the blood glucose response; however, its effect on gastric emptying and its relative mechanism have not been thoroughly investigated. By using Fourier transform infrared (FT-IR), X-ray diffraction (XRD), scanning electron microscopy (SEM), antioxidant ability, and enzymic inhibitory tests for the characterization and in vitro semi-dynamic digestion of complexes of OBG (high and low molecular weights) and sea buckthorn flavonoids, we found that the higher molecular weight complex (FU) exhibited stronger ABTS and DPPH radical scavenging abilities and higher α-glucosidase and α-amylase inhibition rates. Mice fed with rice flour with FU addition exhibited the slowest gastric emptying and intestinal propulsion rates and blood glucose rise and had the lowest activity of digestive enzymes and levels of insulin, ghrelin, motilin (MTL), and relevant gene (ghrelin and GHSR mRNA) expression than those in the control and low-molecular-weight groups. This study provided scientific data for the development of foods with delayed gastric rate and hypoglycemic index for specific populations.

Oxidative Release of Natural Glycans: Unraveling the Mechanism for Rapid N-Glycan Glycomics Analysis.

N-glycosylation is a critical post-translational modification involved in various biosynthetic pathways and disease mechanisms. In this study, we present an optimized oxidative release of natural glycans (ORNG) method using household bleach that enables the rapid and efficient release of N-glycans from biological samples. We thoroughly investigated the ORNG mechanism, identifying key intermediates and side products and providing valuable insights into the oxidative release process. The method is highly efficient, releasing a wide range of N-glycans, including high-mannose, hybrid, and complex structures, with minimal sample processing. Our ORNG-based specific N-glycan profiling approach has demonstrated high sensitivity and efficiency, particularly in releasing N-glycans resistant to enzymatic digestion, such as core α3-fucosylated N-glycans from soy protein. Validation through mass spectrometry confirmed the method's ability to accurately profile N-glycans from complex biological samples, including human serum, with results comparable to traditional PNGase F digestion. The ORNG-based method's scalability, versatility, and use of low-cost reagents make it especially suited for large-scale glycomics studies. Furthermore, the mass spectrometry data revealed that the ORNG-based method achieves high sensitivity and specificity, positioning it as a robust alternative for comprehensive glycan profiling and functional studies. Our findings highlight ORNG's potential to advance N-glycomics, offering promising improvements in speed, efficiency, and breadth of glycan analysis.

Quercetin: Potential antidiabetic effects through enzyme inhibition and starch digestibility

AbstractDiabetes mellitus involves high blood sugar levels due to insufficient insulin action. Furthermore, enzymes such as α‐amylase and α‐glucosidase break down carbohydrates into glucose, leading to postprandial hyperglycemia. Flavonoids, particularly quercetin, inhibit these enzymes, slowing carbohydrate digestion and reducing glucose absorption. Quercetin has significant hypoglycemic effects with inhibitory concentration (IC50) values comparable to acarbose, a standard inhibitor, suggesting its potential as a natural alternative for diabetes management. In silico models, including molecular docking, molecular dynamics (MD) simulations, and quantitative structure‐activity relationship (QSAR) approaches, help researchers understand the molecular interactions of therapeutic agents. These techniques identify potential inhibitors, determine enzyme‐inhibitor structures, and calculate binding energies, correlating findings with in vitro or in vivo data. Molecular docking predicts molecular orientations, MD simulations offer insights into enzyme–inhibitor dynamics, and QSAR models predict inhibitory potential based on structural properties. Studies have shown that quercetin effectively inhibits α‐glucosidase and α‐amylase by forming hydrogen bonds with specific amino acid residues. Quercetin interacts with starches and reduces their digestibility, increases the formation of resistant starch, lowers the glycemic index, and inhibits digestive enzymes. Studies show that the effects of quercetin on starch digestion vary with concentration and type of starch, and its incorporation into foods such as bakery products, pasta, etc. can significantly decrease starch hydrolysis. The incorporation of quercetin into starch matrices may aid in the development of functional foods aimed at improving glycemic control.

Probiotic Strain Lactobacillus plantarum GCLP4 Derived From the Gut of Ctenopharyngodon idella Modulates Growth, Digestive Enzymes and Immune-Biochemical Parameters of Labeo rohita Fingerlings.

The present study was carried out to isolate, identify, and characterize bacterial probiotic strain from the gut of Ctenopharyngodon idella (Grass carp) and then to study its effect on growth, digestive enzymes and immunity of Labeo rohita fingerlings. A total of 6 gram-positive bacteria belonging to the genus Lactobacillus spp. (3), Bacillus spp. (2) and Staphylococcus spp. (1), were identified biochemically. Based on the biochemical results, the isolate GCLP4 was selected for molecular confirmation and BLAST analysis showed maximum homology with Lactobacillus plantarum (100% ident). Fish were fed for 60 days with diet containing 0 (T0), 105 (T1), 107 (T2), 109 (T3) cfu/g diet of L. plantarum GCLP4 and 105 (T4) of commercial probiotics. Results shows that supplementation of GCLP4 had significantly (p < 0.05) improve weight gain (%), specific growth rate (SGR) and feed conversion ratio (FCR) of L. rohita with better values in T3 group. The immunological parameters (white blood cell, red blood cell, haemoglobin, total protein, albumin and globulin were significantly higher (p < 0.05) with 107 and 109 Lactobacillus GCLP4 cfu/g diet. The digestive enzyme activities (protease, amylase and lipase) were significantly (p < 0.05) higher, particularly, with 109 Lactobacillus GCLP4 cfu/g of diet. All the groups supplemented with GCLP4 including the commercial probiotics have lower (p < 0.05) activities of serum transaminase enzymes along with lower (p < 0.005) level of glucose as compared to the control group. The results of the study collectively suggest that dietary L. plantarum GCLP4 at 109 cfu/g is an effective probiotic obtained from grass carp having potency to promote growth, digestive enzymes and immune-biochemical indices of L. rohita fingerlings in present culture condition.

Detection of Babesia spp., and Theileria spp., in sheep across diverse provinces of Iran

This study aimed to identify the diversity of Babesia and Theileria species in sheep across various regions of Iran using microscopic and molecular techniques, including species-specific PCR and enzymatic digestion. A total of 373 blood samples were collected from sheep during the tick vector activity period, from 2018 to 2021, in provinces such as Tehran, Alborz, Qazvin, Hamedan, West Azerbaijan, Kerman, and Fars.Results showed that 101 samples (27 %) exhibited piroplasms including 78 samples of Theileria spp. and 23 samples of Babesia spp. A molecular approach using general primers detected piroplasm parasites in 145 samples (38 %). Theileria ovis was notably present in 91 samples (24.39 %), followed by Theileria lestoquardi in 24 samples (6.43 %). Babesia ovis infection was detected in 30 samples (8.4 %). Despite extensive molecular evaluation, no other Babesia species, including Babesia motasi, were identified. Co-infections involving T. ovis and T. lestoquardi (4 samples; 1.07 %) and T. ovis and B. ovis (6 samples; 1.60 %) were observed. No Babesia spp. were detected in Kerman and Fars provinces, although T. ovis and T. lestoquardi were present.Blast analysis of the sequences indicated 100 % intra-species similarity, with inter-species similarities of 83.3 % (B. ovis and T. lestoquardi), 84.4 % (B. ovis and T. ovis), and 96.44 % (T. ovis and T. lestoquardi). In conclusion, B. ovis was the main cause of Babesiosis, while Theileriosis was predominantly attributed to T. ovis and T. lestoquardi. Molecular diagnostics play a key role in accurately distinguishing between these species, particularly in cases of co-infection involving Babesia spp. and Theileria spp.

Production of Bioactive Peptides from Tartary Buckwheat by Solid-State Fermentation with Lactiplantibacillus plantarum ATCC 14917.

Buckwheat is a valuable crop that contains various nutrients and functional components. Tartary buckwheat peptide is a protease-hydrolyzed protein with a wide range of physiological functions. Tartary buckwheat peptide produced through microbial fermentation can decrease the enzymatic digestion of buckwheat protein, which contributes to the bitter taste, and improve both the flavor and texture of buckwheat peptide products. In this study, microbial fermentation using probiotics was employed to prepare Tartary buckwheat peptides, and the preparation process was optimized. Based on single-factor experiments, the polypeptide content in the fermentation solution initially increased and then decreased with varying water content, inoculum concentration, glucose addition, fermentation temperature, fermentation time, and potassium dihydrogen phosphate addition. According to the response surface methodology, the maximum peptide content was achieved under fermentation conditions of 60.0% moisture content, 12.87% inoculum ratio, 2.0% glucose, and a fermentation temperature of 30.0 °C, with an actual value of (22.18 ± 1.02) mg/mL. The results show that fermentation with Lactiplantibacillus plantarum produces higher peptide levels and is safer than other microbial fermentation methods.

Growth, biochemical indexes, and intestinal microbiota response of red tilapia (Oreochromis spp.) under different densities and feeding frequencies in the land-based aquaculture tank

Red tilapia (Oreochromis spp.) has been favored by farmers in recent years because of its superior growth performance and competitive market price, but there is a lack of experimental data related to growth and health of red tilapia in land-based aquaculture tanks. In this study, red tilapia (initial weight 27.62 ± 2.43 g) was cultured with three stocking densities (30, 40, or 50 fish/m3, named 30F,40F and 50F) in the culture mode for 60 days, including two feeding frequencies (2 or 3 times/day, named 2 T and 3 T), and various growth and metabolic parameters were assessed along with the intestinal microbiome analysis. The 30 fish/m3 groups achieved the better weigh gain rate and specific growth rate (p < 0.05). The growth rate and condition factor of 40F groups were close to those of 30F groups, the yield of 40F groups were not significantly different from that of 50F groups (p > 0.05), and it had the highest survival rate (p < 0.05). The 3 T groups has a higher growth rate and yield but also has a higher rate of feed conversion (p < 0.05). The 30F groups had higher lipase levels, but a higher fat content in this group, more lipids were observed in the liver, and resulted in higher blood glucose levels (p < 0.05). The 40F groups showed healthier physiological and biochemical indicators, as its total cholesterol, lysozyme and antioxidant enzymes levels were similar to those of the 30F groups (p > 0.05), alkaline phosphatase, amylase and trypsin levels were higher than those of the other groups, and aspartate transaminase levels were the lowest (p < 0.05). The 50F groups had higher cortisol, alanine transaminase and malondialdehyde levels, lower lysozyme and digestive enzyme activities, and blood cell infiltration in the liver (p < 0.05). In terms of different feeding frequencies, the 3 T groups had higher crude protein content, catalase and total superoxide dismutase levels, wider villus width and lower blood glucose level, while the 2 T groups had higher triglyceride, total cholesterol, lysozyme, alkaline phosphatase and some digestive enzymes activities (p < 0.05). From the perspective of intestinal microbiota. The 40F groups had a better intestinal flora, and 40F-3T groups observed a better balance of flora, while 40F-2T groups had more digestion-related flora. Considering the growth rate, health status, and yield, the optimum culture condition was determined to be stocking density of 40 fish/m3, and feedings 3 times/day were recommended. The experiment provided theoretical basis and experimental data for red tilapia culture in land-based aquaculture tank mode.