Impact of Dietary Supplementation of Flaxseed Meal on Intestinal Morphology, Specific Enzymatic Activity, and Cecal Microbiome in Broiler Chickens

The use of natural feed additives could be a beneficial approach to maintaining the health of chickens and a way to improve food digestion. Flaxseed is a rich source of omega-3 fatty acid, alpha linolenic acid, oleic acid, and fiber. The purpose of this study was to evaluate the effects of dietary inclusion of 4% flaxseed on the intestinal morphology, specific enzymatic activity, and cecal microbiome in broiler chickens. The 4-week feeding trial was conducted on 100 Cobb 500 (14 days of age) unsexed broiler chickens divided into two groups: a control group (C) and an experimental group (E). The broilers were housed in boxes of size 3 m2 (each group was housed in a single box with 10 replicates, 5 chickens per replicate) and reared on permanent wood shaves litter (10–12 cm thick). At the end of the experiment, chickens (n = 10) were sacrificed and tissue samples were harvested from the duodenum, jejunum, and cecum for histological, enzymatic, and microbiome analyses. In group E, histological analysis revealed a significant increase in villus height (p < 0.001) possibly leading to enhanced intestinal nutrient absorption. An increase in the specific activities of α-amylase (p < 0.05), invertase (p < 0.01), and endo-β-1,4-glucanase (p < 0.001) was noticed in the E group for the duodenum and jejunum compared to the control group. In contrast, maltase activity decreased in the duodenum and increased in the jejunum in the E group. The trypsin and lipase specific activities did not vary in a significant way. In addition, the cecal microbiome of the E group was characterized by an increase in Lactobacilli (p < 0.01) and Clostridium coccoides and a decrease in Bacteroides, Ruminoccocus, Enterobacteriaceae, and Clostridium leptum. In conclusion, our results suggest that dietary supplementation of flaxseed meal may boost intestinal health status in poultry.

Germination of Crotalaria and Lupinus (Fabaceae) seeds submitted to different pre-germination treatments and their effect on enzymatic activity during early germination

Most of the wild and native legume seeds has a hard and impermeable testa, which causes physical dormancy and prevents them from germinating even when environmental conditions are favorable. The study evaluated the effect of scarification treatments on germination and enzymatic activity of Crotalaria longirostrata (Cl) and Lupinus exaltatus (Le) seeds. After scarification treatments, germination percentage (GP) and rate (GR) were assessed during 30 days after seeding (DAS); and water absorption (WA) and specific enzymatic activity (SEA) during early germination (0, 6, 18, 36, 72, 120 h) in a growing chamber at 25 °C and photoperiod of 12 h. Scarification with 98% H2SO4 15 min increased GP and GR in both species. At 30 DAS, GP and GR of Le seeds were 34% and 0.97 seeds day-1, respectively. In Cl seeds, GP was 64% and GR 0.90 seeds day-1. Scarification with H2O at 80 °C 1 min also promoted germination in Cl (52%). At 120 h after seeding, Le and Cl seeds showed already a high GP with acid scarification (31% and 48%, respectively). In seeds of both species, scarification treatments affected WA and SEA during early germination. During this period, scarification treatments that increased GP also showed a higher α-D-galactosidase activity. The maximum enzyme activity was observed 72 h after hot water scarification in Cl (82.6 U/mg total protein), followed by acid scarification (54.5 U/mg total protein). In Le, the activity peak was 36 h after acid scarification (9.5 U/mg total protein). No relationship was observed between β-glucosidase activity and GP in both species. In conclusion, during early germination of both species, the increase in GP is accompanied by a rise in α-D-galactosidase activity between 36 and 72 h after seeding; and in Cl seeds, an alternative scarification treatment to increase GP may be the use of hot water.

Detection of enzymes produced by lactic acid bacteria isolated from traditionally made Serbian cheese and their role in the formation of its specific flavor

Nine species (sixteen isolates) of lactic acid bacteria (LAB) isolated from traditionally made Serbian cheese were evaluated for their enzymatic activities in order to select indigenous strains of technical interest for the manufacture of cheese. These strains were selected based on their previously determined biochemical and physiological characteristics, as well as their antimicrobial activity, and were identified as Lactococcus lactis subsp. lactis (one isolate), Lc. lactis subsp. lactis biovar. diacetylactis (five isolates), Lactobacillus fermentum (two isolates), Lb. plantarum (one isolate), Lb. brevis (one isolate), Enterococcus faecalis (three isolates), E. faecium (one isolate), E. durans (one isolate) and E. hirae (one isolate). The enzymatic activities (acid and alkaline invertases, alkaline phosphatase, alkaline protease, a-amylase) were measured by using the spectrophotometric method. The results indicated that all Lactobacillus isolates showed protease, amylase, and alkaline phosphatase activities, while the activities of acid and alkaline invertases were not observed. The Lactococcus isolates showed protease, acid invertase and alkaline phosphatase activities, except the KGPMF50 isolate, which showed no alkaline phosphatase activity. The tested Enterococcus isolates showed weakly and strain-specific enzymatic activity. The results indicated that the enzymes produced by the investigated strains have a role in the formation of the specific flavor of cheese and that these isolates, especially Lactobacillus isolates, showed the potential for use in the dairy industry or applied biotechnology.

Specific Enzymatic Activity of Ceruloplasmin as a Potential Indicator of Copper Status

Background:: The serum copper (Cu) and ceruloplasmin (Cp) concentrations are common blood markers of copper metabolism. In altered physiological conditions, Cp can act as an acute phase reactant and its concentration may increase. Objective:: To evaluate specific enzymatic activity of Cp as a potential indicator of Cu status and its correlation with serum Cu level. Methods:: Serum Cu levels were estimated as per NIOSH method. Specific enzymatic activity of Cp was determined from enzymatic activity and immune concentration of Cp as per standard methods. The statistical analysis was carried out using the package of social science (SPSS) software. Results:: The difference in mean specific enzymatic activity of Cp was statistically significant between clinical and control groups. In control population, the correlation between serum Cu level and specific enzymatic activity of Cp was moderate and statistically significant (r=0.566, p=0.014, N=18) as compared to the clinical group (r=0.338, p=0.016, N=50). Conclusion:: The study revealed that clinical group was significantly different in specific enzymatic activity of Cp as compared to control group. Besides this, the specific enzymatic activity of Cp was moderately but significantly correlated with serum Cu level in control group but did not reveal conclusive evidence in clinical population.

Vpr and Its Cellular Interaction Partners: R We There Yet?

Vpr is a lentiviral accessory protein that is expressed late during the infection cycle and is packaged in significant quantities into virus particles through a specific interaction with the P6 domain of the viral Gag precursor. Characterization of the physiologically relevant function(s) of Vpr has been hampered by the fact that in many cell lines, deletion of Vpr does not significantly affect viral fitness. However, Vpr is critical for virus replication in primary macrophages and for viral pathogenesis in vivo. It is generally accepted that Vpr does not have a specific enzymatic activity but functions as a molecular adapter to modulate viral or cellular processes for the benefit of the virus. Indeed, many Vpr interacting factors have been described by now, and the goal of this review is to summarize our current knowledge of cellular proteins targeted by Vpr.

Biosynthesis of silver nanoparticles employing Trichoderma harzianum with enzymatic stimulation for the control of Sclerotinia sclerotiorum

Biogenic synthesis of silver nanoparticles employing fungi offers advantages, including the formation of a capping from fungal biomolecules, which provides stability and can contribute to biological activity. In this work, silver nanoparticles were synthesized using Trichoderma harzianum cultivated with (AgNP-TS) and without enzymatic stimulation (AgNP-T) by the cell wall of Sclerotinia sclerotiorum. The nanoparticles were evaluated for the control of S. sclerotiorum. The specific activity of the T. harzianum hydrolytic enzymes were determined in the filtrates and nanoparticles. Cytotoxicity and genotoxicity were also evaluated. Both the nanoparticles exhibited inhibitory activity towards S. sclerotiorum, with no new sclerotia development, however AgNP-TS was more effective against mycelial growth. Both the filtrates and the nanoparticles showed specific enzymatic activity. Low levels of cytotoxicity and genotoxicity were observed. This study opens perspectives for further exploration of fungal biogenic nanoparticles, indicating their use for the control of S. sclerotiorum and other agricultural pests.

Proteolytic Activity of Edible Spruce Morchella esculenta

This study focused on the determination of non-specific proteolytic activity of edible spruce Morchella esculenta in water extract, phosphate-buffered saline (PBS) solution (pH = 7.5) extract and a suspension prepared from 200 mg DW (dry weight) of edible spruce in PBS solution (pH 7.5). A clear casein solution was used as a substrate. The absorbances were measured in quartz cuvettes at the wavelength of 280 nm against a blank with zero concentration of trypsin. Non-specific proteolytic activity was expressed as trypsin equivalents per kilogram of mushroom dry weight (mg.kg−1 DW). All of the extracts demonstrated non-specific enzymatic activity. The highest activity was observed in the PBS suspension and the lowest enzymatic activity was measured in the water extract of the Morchella esculenta fungi. The non-specific proteolytic activity decreased in the following order: PBS suspension extract (pH 7.5; 22.9 mg.kg−1 DW), followed by PBS extract (pH 7.5; 13.6 mg.kg−1 DW) and finally the water extract (10.94 mg.kg−1 DW).

Impact of cadmium on the level of hepatic metallothioneins, essential elements, and selected enzymes in the experimental rat model

The response of different strains of laboratory rats (Rattus norvegicus L.) on both acute (via intraperitoneal injection) and chronic (via drinking water and/or diet) cadmium intoxication was investigated in the model study. The rat strains Long Evans (LE), Spontaneously hypertensive rat (SHR), and Brown Norway (BN) were tested and compared, and total Cd levels and metallothionein (MT) concentrations were determined in the liver of experimental animals. The liver MT concentrations were determined by using adsorptive chronopotentiometry and modified Brdička reaction and were significantly correlated (r = 0.965) with the total liver Cd content. Moreover, the Cd application resulted in increasing zinc liver contents confirming intensive MT synthesis in the rat liver. In the blood plasma, specific enzymatic activity of glutathione reductase (GR) and glutathione-S-transferase (GST) was determined suggesting increasing activity of GR with the amount of applied Cd for all three strains, whereas ambiguous results have been found for the activity of GST. Therefore, MT concentrations seemed to be more sensitive indicators of the Cd intoxication compared to the assessment of the specific enzymatic activity. &nbsp;

Production of Recombinant β-Galactosidase in Lactobacillus plantarum, Using a pSIP-Based Food-Grade Expression System

Food-grade expression systems based on using food-grade microorganisms have been developed for the production of recombinant enzymes used in food applications. Lactic acid bacteria (LAB), especially Lactobacilli, have been widely used for various purposes in food and recognized as a promising host of food-grade enzyme production. In this study, the pSIP409 vectors, originally containing the erm gene, were used to replace this selection marker by the alr gene resulting in the production of the pSIP609 expression vector in L. planatarum. This vector could express high amounts of β-galactosidases, showing both high volumetric as well a specific enzymatic activity. Thus, the food-grade recombinant enzyme production in L. planatarum harboring pSIP609 was very fruitful and useful for food industries.