Lysozyme Concentration Research Articles

Effect of continuous flow HTST treatments on donkey milk nutritional quality

Nutritional quality of raw donkey milk (DM) may be impaired during sanitization with the current batch holder pasteurization systems (62.5 °C for 30 min). In this paper, we present the preliminary results concerning the effects of high temperature for short time (HTST) protocols using an innovative in continuous low flow rate pasteurization plant (60 dm3/h) on B-vitamins group, antioxidant capacity, lysozyme and β-lactoglobulin in DM. Lysozyme, β-lactoglobulin and antioxidant power decreased after the thermal treatments, with characteristics depending on the extent of the heat treatment. The lysozyme content was substantially reduced between 20 and 60%, while the degradation of β-lactoglobulin was lower (2–22%). No vitamin B1 and B12 were found in raw milk, whereas were detected vitamin B2 (0.17 μmol/L), nicotinic acid (13.28 μmol/L), B6 (2.06 μmol/L) and B9 (0.75 μmol/L). The heat treatments carried out with the innovative plant ensured vitamin retention, as no significant differences were found against the raw milk (p > 0.05). The preliminary results from this study represent a guidance to the establishment of DM pasteurization standards parameters with the perspective to improve DM nutritional quality.

Nutrients and Bioactive Components of Human Milk After One Year of Lactation: Implication for Human Milk Banks.

Specifying the nutrient content and bioactive compounds in milk from long-term lactation for the purpose of finding additional sources of donors' milk for human milk banks. Human milk samples were collected from 43 mothers of term infants (term infant human milk, TIHM) (3-6 weeks of lactation) and 50 mothers who have breastfed for over a year (long-nursing human milk, LNHM). The milk collection time was 24 hours. The analyses of fat, protein, carbohydrate and energy content were performed with human milk analyzer MIRIS (Miris HMA™ ); lactoferrin and vitamin C content using high performance liquid chromatography in reversed phase (RP-HPLC); total antioxidant activity (TAC) and lysozyme by enzyme-linked immunosorbent assay (ELISA); and the activity of glutathione peroxidase (GPx) and catalase (CAT) via spectrophotometric methods. Sociodemographic characteristics of both groups of mothers have been compiled. Higher fat content and energy value was found in LNHM than in TIHM. Protein content in LNHM and TIHM did not differ. Carbohydrate content was lower in LNHM than in TIHM. TAC, CAT and GPx activity were higher in LNHM compared to TIHM. No significant differences in the content of lysozyme, lactoferrin and vitamin C between the studied groups were found. The findings suggest that after 12 months of lactation, human milk still has significant nutritional value, does not lose bioactive components and can be considered for use in human milk banks.

Effect of Feed Containing Hermetia illucens Larvae Immunized by Lactobacillus plantarum Injection on the Growth and Immunity of Rainbow Trout (Oncorhynchus mykiss).

Simple SummaryIn this study, we evaluated the effect on the growth and immunity of rainbow trout of a feed formulated using Hermetia illucens with increased antimicrobial peptides expression by Lactobacillus plantarum infection (ImHIL). As a result, growth and immunological indicators improved, and therefore, ImHIL is expected to become a good feed source for rainbow trout aquaculture.We investigated the effects of a feed containing Hermetia illucens larvae injected with bacteria on the growth and immunity of Oncorhynchus mykiss. The feed was prepared by replacing fishmeal in feed with 25 and 50% nonimmunized (HIL25, HIL50) or immunized HIL (ImHIL25, ImHIL50), and its protein:fat:carbohydrate ratio was 45:15:18. ImHIL extracts showed inhibitory activity against fish pathogenic bacteria. Both red blood cell count and insulin-like growth factor-1 as the growth indicator were the highest among the groups at week 6 after feeding in the ImHIL50 group. As immune indicators, blood aspartate aminotransferase levels were lower in the ImHIL25 and ImHIL50 groups than in that of other groups at week 6 after feeding, and lysozyme content was significantly higher in ImHIL25 and ImHIL50. The above results demonstrate that ImHIL has a beneficial effect on the improvement of growth and immunity. Accordingly, we suggest that ImHIL has the potential to be a good feed source in aquaculture.

Naringenin Regulates Lipopolysaccharide-Induced Abnormal Airway Surface Liquid Secretion

Airway surface liquid (ASL) is one of the key factors affecting the respiratory system's physiological function. Abnormal ASL secretion can increase the incidence of various respiratory diseases. Lipopolysaccharide (LPS) stimulation can damage the airway epithelial barrier, affect the concentration of ASL contents, and down-regulate ion channel expression, which in turn causes abnormal ASL secretion. Naringenin, which exists in many Citrus foods, has the ability to promote airway surface liquid secretion. This work is designed to investigate the regulatory mechanism of naringenin on LPS-induced abnormal ASL secretion. The effects of naringenin and LPS on the viability of Calu-3 cells were measured by CellTiter 96® AQueous One Solution Cell Proliferation Assay (MTS). ASL secretion volume was measured by a micropipette on air–liquid interface cultured cells. The concentration of Cl−, Na+, lysozyme, and total protein in ASL were respectively measured by assay kits. The mRNA expressions were determined by quantitative real-time polymerase chain reaction, and proteins were measured by enzyme-linked immunosorbent assay. The results indicated that LPS could affect ASL secretion and regulate cystic fibrosis transmembrane conductance regulator (CFTR), aquaporin 1 (AQP1) and aquaporin 5 (AQP5) expression. Naringenin had the ability to regulate the ASL secretion by increasing secretion volume, and Cl− and Na+ concentrations, reducing lysozyme and total protein content, and regulating CFTR, AQP1, and AQP5 expression. This study indicated that naringenin had regulating effects to attenuate LPS-induced abnormal ASL secretion.

Competitive binding of synergistic antioxidant chlorogenic acid and (−)-epigallocatechin gallate with lysozyme: Insights from multispectroscopic characterization, molecular docking and activity evaluation

The individual and combined binding of two active polyphenols, (−)-epigallocatechin gallate (EGCG) and chlorogenic acid (CGA), to lysozyme was studied at the molecular level by multispectroscopic techniques and molecular docking. The thermodynamic parameters obtained from fluorescence data indicated that the main driving force for the binding of CGA to lysozyme was electrostatic interaction while the binding of EGCG was mainly driven by hydrogen bonding and van der Waals’ forces. The results of circular dichroism spectroscopy and dynamic light scattering showed that the α-helical content and particle size of lysozyme decreased and increased, respectively, in the presence of EGCG and/or CGA. The competitive binding of EGCG and CGA to lysozyme was confirmed by molecular docking. The binding of EGCG and/or CGA inhibited the activity of lysozyme. The combination of EGCG and CGA had synergistic antioxidant activity and cytotoxicity to HeLa cells. Response surface methodology was used to analyze the maximum synergistic antioxidant activity between EGCG and CGA. The lysozyme + EGCG/CGA interaction weakened and enhanced the antioxidant activity and cytotoxicity of EGCG/CGA, respectively. These results would provide ideas for the combined application of EGCG and CGA in the fields of functional foods and pharmaceuticals.

Supplementing Chitosan Oligosaccharide Positively Affects Hybrid Grouper (Epinephelus fuscoguttatus ♀ × E. lanceolatus ♂) Fed Dietary Fish Meal Replacement With Cottonseed Protein Concentrate: Effects on Growth, Gut Microbiota, Antioxidant Function and Immune Response

Oligosaccharides have recently received much attention from researchers owing to their multiple biological activities. This study was conducted to investigate the effects of a diet with reduced fish meal and chitosan oligosaccharide (COS) supplementation on a hybrid grouper (Epinephelus fuscoguttatus ♀ × E. lanceolatus ♂). Seven isonitrogenous and isolipidic types of diet were formulated to feed the groupers for 56 days. To prepare the feed, a reference diet (FM group) containing 55% fish meal was used. Concentrated cottonseed protein (CPC) was used to replace 45% of the fish meal protein, and different COS supplementation levels (0, 0.2%, 0.4, 0.6, 0.8, and 1%) were added. After an 8-week breeding trial, Vibrio harveyi bacteria were injected into the groupers for a 7-day challenge test. The results showed that the FM and COS0.4 groups showed the best growth performance among the groups (p < 0.05); however, there was no significant difference in the survival rate (p > 0.05). Unlike in the FM group, adhesion and breakage of the intestinal plica occurred in the COS0 group. The height and width of the gut fold reached maximum values in the COS0.4 group (p < 0.05). Microbiome sequencing suggested that there was a stable microbiota core in the gut of the groupers. With increasing COS levels, the abundance of both beneficial bacteria and conditional pathogens increased; the activities of serum glucose oxidase, catalase, and total superoxide dismutase also increased (p < 0.05). In the gut tissue, the activities of glutathione peroxidase, glutathione reductase, and glutamine increased first but then decreased (p < 0.05); the contents of lysozyme, acid phosphatase, complements C3 and C4, and IgM showed upward trends (p < 0.05). Compared with that in the FM group, the expression of IL-1β and TNF-α in the COS0 group was upregulated. Gene expression levels of TLR22, TGF-β, and Nrf2 increased first but then decreased with COS supplementation levels (p < 0.05). COS supplementation reduced the cumulative mortality of the groupers in the challenge test (p < 0.05). In general, the results of this study demonstrated that dietary COS supplementation enhanced growth performance, intestinal health, and antioxidant and immune responses of groupers fed with a low-fish meal diet. The optimal and acceptable levels of COS supplement were 0.45 and 0.4–0.6%, respectively; these values can provide a reference for developing aquatic prebiotics.

Transcriptome Atlas of 16 Donkey Tissues.

Donkeys (Equus asinus) are important livestock with great economic value in meat, skin, and milk production. However, a lack of knowledge of the transcriptome landscape across a wide range of donkey tissues limits genetic selective breeding and conservation. Here we used transcriptomics to describe the transcriptome landscape, classify the tissue-specific gene expression across all primary donkey tissues, and present supplementary analyses on the protein level of additional donkey milk samples. Overall, 16,013 protein-coding genes and 21,983 transcripts were mapped to the reference genome, including 6,778 ubiquitously expressed genes and 2,601 tissue-enriched genes. Functional analysis revealed that the function of the tissue-enriched genes was highly tissue specific. Tissue-elevated genes that could be associated with unique phenotypes in donkey were analyzed. The results showed that, compared with those in human and other livestock, the lysozyme gene in donkey breast was specifically and highly expressed. The calcium-binding lysozyme, encoded by the lysozyme gene, was also detected in high amounts in donkey milk. Given those intact lysozyme genes that predict potentially functional calcium-binding lysozyme found in only a few species (e.g., donkey and horse), the high expression of the lysozyme gene in donkey breast may contribute to the high lysozyme content in donkey milk. Furthermore, 71% of the proteins in donkey milk overlapped with human milk protein, higher than the overlapping rates of bovine, sheep, and swine with humans. The donkey transcriptomic resource contributes to the available genomic resources to interpret the molecular mechanisms underlying phenotype traits.

Evaluation of an oral DNA nanovaccine against photobacteriosis in Solea senegalensis

Infectious diseases are one of the main causes of social and economical losses in world aquaculture. Senegalese sole (Solea senegalensis) is an important species for aquaculture in southern Europe, whose production is affected by the appearance of bacterial diseases such as photobacteriosis, a septicemia caused by Photobacterium damselae subsp. piscicida (Phdp). The aim of this study was to obtain an oral DNA nanovaccine and to evaluate its efficacy against Phdp in S. senegalensis juveniles. For this purpose, the amplified product corresponding to the protein inosine-5′-monophophate dehydrogenase (IMPDH) from Phdp, was cloned into the expression vector pcDNA™6.2/C-EmGFP-GW obtaining the DNA vaccine named as pPDPimpdh. The correct transcription and protein expression was verified at 48 h post tansfection in HEK293 cells. Chitosan nanoparticles (CS-TPP NPs) were prepared by ionotropic gelation and their features were appropriate for use as oral delivery system. Therefore, pPDPimpdh was protected with chitosan CS-TPP NPs throughout complex coacervation method giving as a result a DNA nanovaccine referred as CS-TPP+pPDPimpdh NPs. Sole juveniles were vaccinated orally with CS-TPP NPs, pPDPimpdh and CS-TPP+pPDPimpdh NPs followed by a challenge with Phdp at 30 days post vaccination (dpv). The relative percentage survival (RPS) for pPDPimpdh vaccinated groups was 6.25%, probably due to its degradation in the digestive tract. RPS value obtained for CS-TPP NPs and CS-TPP+pPDPimpdh NPs was 40% and antibodies were observed in both cases. However, a delay in mortality was observed in sole juveniles vaccinated orally with CS-TPP+pPDPimpdh NPs. In fact, an upregulation of tf, mhcII, cd8a and igm in the posterior gut and c3, hamp1, tf and cd4 in spleen was observed in juveniles vaccinated with CS-TPP+pPDPimpdh NPs. After challenge, a modulation of cd8a and cd4 expression levels in the posterior gut and c3, tf, lyg, cd4, igm and igt expression levels in spleen was observed. Moreover, the concentration of lysozyme in skin mucus significantly increased in fish vaccinated orally with CS-TPP+pPDPimpdh NPs at 11 dpc. These data indicate that oral vaccination with CS-TPP+pPDPimpdh NPs could be acting through the non-specific immune responses as well as the specific humoral and cell mediated immunity and provide the first step toward a development of an oral DNA nanovaccine against Phdp in sole.

Effects of the prebiotic lactoferrin on multidrug-resistant Escherichia coli infections in broiler chickens.

Background and Aim:Increased multidrug resistance in Escherichia coli has created challenges for the poultry industry. Consequently, new antimicrobial agents should preferentially be utilized for the prevention and treatment of E. coli outbreaks. This study aimed to evaluate the effects of lactoferrin (LF) as a prebiotic on broiler chicks challenged with multidrug-resistant E. coli in comparison with antibiotics.Materials and Methods:A total of 70 diseased flocks from Egypt were collected for E. coli isolation and identification, serotyping, and antimicrobial susceptibility pattern determination. E. coli was isolated and characterized phenotypically and one isolate that showed multidrug-resistance was selected. A challenge trial was performed to evaluate the effectiveness of LF as a prebiotic on the isolated multidrug-resistant E. coli. Liver samples were collected from the experimental chicks and subjected to E. coli enumeration to illustrate the effectiveness of LF on the liver cells and bacteria using an electron microscope. Serum samples were also collected to estimate lysozyme and nitric oxide (NO) concentrations.Results:After isolation of E. coli with a percentage of 54.3% from the diseased broilers, the strain was serotyped (identified serotypes: O2, O18, O55, O78, O86a, O111, O125, O126, O127, O157, O159, and O166). Multi-antibiotic resistance was found to be harbored in a high percentage among 11 antibiotic discs. The LF in the prophylactic and treated groups was found to have a significant effect in comparison with the group treated with the drug of choice (ciprofloxacin). Furthermore, a significant difference in the NO (one of non-specific immune response) and a non-significant difference in lysozyme concentrations were reported in the group fed on rations with LF in comparison with the non-fed group.Conclusion:LF was thus identified as an effective prebiotic that can improve chick performance, help them to overcome multidrug-resistant E. coli and stimulate immunity.

Sensitivity Enhancement in the Colorimetric/Spectroscopic Determination of Lysozyme Concentration in Nanomolar Level with Colloidal Citrat e Capped Au@Ag Core-shell Nanoparticles

Background: Lysozyme level in body fluids is a significant indicator of various diseases. Cheap and simple colorimetric detection of lysozyme, in biological sample, by gold and silver nanoparticles is a field of interest of nanoparticle research for more than a decade. Objective: We report here an attempt to improve the sensitivity part of the colorimetric lysozyme detection process by using citrate capped gold-silver core-shell nanoparticles without any functionalization. Methods: Performance of gold-silver core-shell nanoparticles in determining of lysozyme concentration in water is done using colorimetric/spectroscopic technique. Gold nanoparticles, silver nanoparticles and mixed colloidal suspension of gold and silver nanoparticles, for comparison have also been studied. Selectivity study has been performed through spectroscopic analysis, red-greenblue colour component analysis and fractal dimension analysis of the nanoparticles interacted with several low and high isoelectric point proteins. Results: Gold-silver core-shell nanoparticles showed higher sensitivity for a wider range of lysozyme concentration compared to gold and silver nanoparticles prepared by us or reported in literature. Gold nanoparticles showed higher sensitivity compared to the core-shell nanoparticles, but for a narrow concentration range of lysozyme. For silver nanoparticles and the mixed nanoparticle system, both sensitivity and range of determination of lysozyme concentration were much smaller compared to the core-shell nanoparticles. Core-shell nanoparticles showed better selectivity compared to gold nanoparticles in identifying aquatic solution of lysozyme from that of other proteins. Conclusion: Gold-silver core-shell nanoparticles have higher sensitivity in determining wide range of lysozyme concentration in water compared to gold and silver nanoparticles reported in literature.

Physiological and transcriptional analysis of Chinese soft-shelled turtle (Pelodiscus sinensis) in response to acute nitrite stress

Nitrite is a harmful substance in aquaculture, and has a serious impact on the survival of the Chinese soft-shelled turtle, Pelodiscus sinensis. However, the cellular responses of P. sinensis to nitrite stress have not yet been investigated. The present study showed that nitrite led to a decrease in hemoglobin content and an increase in methemoglobin content in the blood, thus reducing the oxygen-carrying capacity of blood in P. sinensis. Nitrite also affects the antioxidant system of the liver and leads to lipid peroxidation. In addition, nitrite caused immune responses, including a decrease in lysozyme content and an increase in total complement activity, interleukin-6, and heme oxygenase concentrations in the serum. Additionally, the terminal deoxynucleotidyl transferase-mediated nick end labeling (TUNEL) assay showed that apoptosis occurred in liver cells. Finally, a comparative transcriptome analysis was conducted. A total of 573 differentially expressed genes and 15 significantly enriched KEGG pathways were identified. Among them, the glutathione S-transferase omega 1 (GSTO1) gene may relieve nitrite-induced oxidative damage in P. sinensis by participating in a variety of redox-related pathways, while the PPAR signaling pathway has been proposed to play an important regulatory role in lipid metabolism and immune responses. The present study comprehensively explored the cellular responses of P. sinensis to nitrite stress and provided guidance for future studies.

Investigation of the antibacterial activity against Cutibacterium acnes of lysozyme pu-rified from “Co Co” duck egg whites

Lysozyme has been applied in various fields such as food technology, medicine, and diagnostics because it can resist many types of bacteria. In this research, lysozyme from duck egg whites was studied to evaluate the antibacterial activity against Cutibacterium acnes (C. acnes) often causing acne on human skin. Lysozyme was purified from duck egg whites by ion-exchange chromatography and gel-filtration chromatography. After that, this enzyme was used to investigate the resistance to C. acnes at different pH, temperature, concentration, and storage conditions. The results presented that lysozyme exhibited the best resistance to C. acnes at pH 6.0 and 6.5 on trypticase - yeast extract - heart extract - glycerol agar (TYEG) medium, at 30°C and 35°C. Additionally, these conditions had the least effect on lysozyme antibacterial activity. The minimal inhibitory concentration (MIC80) and minimum bactericidal concentration (MBC) of lysozyme to C. acnes were 0.55 mg/mL and 1.11 mg/mL, respectively. Lysozyme could keep up the best antimicrobial activity when stored at -20oC and -10oC; After 30 days, it still kept nearly 80% of its activity. These findings will offer a basis for larger-scale production of lysozyme powder for further research and commercial purposes, especially skin-care products.

Ocular Surface Pathology in Patients Suffering from Mercury Intoxication.

Purpose: To report the ocular surface pathology of patients suffering from acute/subacute mercury vapor intoxication. Design: Cross-sectional study. Participants: Male workers intoxicated with inorganic mercury referred for ophthalmic involvement and healthy control subjects. Methods: The following tests were performed: dry eye (DE)-related symptoms indicated by the ocular surface disease (OSDI) index questionnaire; tear osmolarity; analysis of 23 tear cytokine concentrations and principal component and hierarchical agglomerative cluster analyses; tear break-up time (T-BUT); corneal fluorescein and conjunctival lissamine green staining; tear production by Schirmer and tear lysozyme tests; mechanical and thermal corneal sensitivity (non-contact esthesiometry); and corneal nerve analysis and dendritic cell density by in vivo confocal microscopy (IVCM). Results: Twenty-two out of 29 evaluated patients entered the study. Most had DE-related symptoms (OSDI values > 12), that were severe in 63.6% of them. Tear osmolarity was elevated (>308 mOsms/L) in 83.4% of patients (mean 336.23 (28.71) mOsm/L). Corneal and conjunctival staining were unremarkable. T-BUT was low (<7 s) in 22.7% of patients. Schirmer test and tear lysozyme concentration were low in 13.6% and 27.3% of cases, respectively. Corneal esthesiometry showed patient mechanical (mean 147.81 (53.36) mL/min) and thermal thresholds to heat (+2.35 (+1.10) °C) and cold (−2.57 (−1.24) °C) to be significantly higher than controls. Corneal IVCM revealed lower values for nerve density (6.4 (2.94) n/mm2), nerve branching density (2 (2.50) n/mm2), and dendritic cell density (9.1 (8.84) n/mm2) in patients. Tear levels of IL-12p70, IL-6, RANTES, and VEGF were increased, whereas EGF and IP-10/CXCL10 were decreased compared to controls. Based on cytokine levels, two clusters of patients were identified. Compared to Cluster 1, Cluster 2 patients had significantly increased tear levels of 18 cytokines, decreased tear lysozyme, lower nerve branching density, fewer dendritic cells, and higher urine mercury levels. Conclusions: Patients suffering from systemic mercury intoxication showed symptoms and signs of ocular surface pathology, mainly by targeting the trigeminal nerve, as shown by alterations in corneal sensitivity and sub-basal nerve morphology.

Inclusion of a protein-rich yeast fraction in rainbow trout plant-based diet: Consequences on growth performances, flesh fatty acid profile and health-related parameters

Fast development of the aquaculture sector in the last decades conducted to a shortage in traditional ingredients (fishmeal and fish oil) that were not anymore avaible in sufficient quantity to purvey such growing aquafeeds needs. Ingredients of plant-derived raw materials, thus started to be preferentially incorporated into aquafeed formulas, and are nowadays being routinely used in most fish feed formulas. However, the total replacement of fishmeal and fish oil has not yet been reached due to negative impacts on feed intake and feed efficiency, perturbations of metabolic and health status, impacts on flesh quality, and growth performances slowdown. Single cell protein sources such as yeast protein products could be an alternative solution to complement and mitigate detrimental effects of plant proteins. This was assessed in a 12 weeks trial in which rainbow trout (Oncorhynchus mykiss) juveniles were fed with a plant-based diet (totally devoid of fishmeal and fish oil), containing 0, 5, 10 or 15% of a protein-rich yeast fraction and microalgae as DHA source.Results indicated that a 15% protein-rich yeast fraction inclusion may upgrade growth performance of trout fed plant-based diets by improving feed conversion ratio and body nitrogen retention. Surprisingly, 5% yeast incorporation in the diet leaded to enhanced accumulation of DHA in whole body that could be linked to a higher expression of the fatty acid elongase 2. Immune status and gut health of fish were also assessed through histological analyses and transcript expression level measurements of immune-related genes in distal intestine complemented by measurement of some plasma immune-related parameters. A lower score of lymphocytes infiltrates in lamina propria of fish distal intestine was observed from 10% inclusion level of the yeast fraction and the calculated overall inflammation score was greatly reduced with 15% inclusion level. In fish fed diets containing 5% and 15% yeast fraction, a higher internal perimeter: external perimeter ratio was measured compared to fish fed the control plant-based group. Inclusion of 15% yeast fraction in the plant-based diet also lead to an increased expression of mucin 5 and claudin a compared to 5% inclusion level. A statistically negative linear correlation was revealed between plasma neutrophils proportions and dietary yeast content. Plasma lysozyme concentration was lower in trouts fed diets containing 10% and 15% yeast fraction. All together, those results revealed that inclusion of yeast in a plant-based diet enables to improve rainbow trout gut health and global immune status.

Effect of Laying Hen Genotype, Age and Some Interior Egg Quality Traits on Lysozyme Content

Abstract The objective of the study was to evaluate the quality of eggs laid by native breed hens of different ages (33 and 53 weeks of age), with particular consideration of the lysozyme parameters. Eggs were collected from 6 breeds/strains of laying hens under the genetic resources conservation in Poland, i.e. Greenleg Partridge (Z-11), Yellowleg Partridge (Ż-33), Rhode Island Red (R-11), Rhode Island White (A-33), Sussex (S-66) and Leghorn (H-22). The yolks of eggs from older hens had a more favourable, intense colour, but the freshness parameters (albumen height and Haugh units) of these eggs were lower. The albumen of eggs from older hens showed greater content and higher enzymatic activity of lysozyme in both thin and thick albumen. Among the six strains of hens, lysozyme content and enzymatic activity were highest in the egg albumen of strains Ż-33, Z-11 and R-11, and lowest in that of strain H-22. The analysed eggs showed statistically significant correlations between some interior egg quality traits (albumen weight, albumen height, Haugh units, yolk colour) and the discussed enzyme parameters. These traits are influenced by genotype and age of the hens.

Dietary bile acid supplementation reveals beneficial effects on intestinal healthy status of tongue sole (Cynoglossus semiliaevis)

The aim of this study was to investigate the effects of dietary bile acids (BAs) on intestinal healthy status of tongue sole in terms of immunity, antioxidant status, digestive ability, mucosal barrier-related genes expression and microbiota. Three experimental diets were prepared with BA levels at 0 mg/kg (CT), 300 mg/kg (BA1) and 900 mg/kg (BA2) in a commercial basal diet. Each diet was fed to three replicates with 120 fish (10.87 ± 0.32 g) in each tank. After an 8-week feeding trial, growth parameters were significantly enhanced in both BAs supplementary groups (P < 0.05), and compared with CT group, survival rate in BA2 group was significantly improved (P < 0.05). Intestinal lysozyme activity and contents of immunoglobulin M and complement 3 were significantly increased in both BAs supplementary groups (P < 0.05), suggesting an enhancement effect on the non-specific immune response. BAs inclusion also significantly improved intestinal antioxidant capabilities by increasing antioxidase activities and decreasing malondialdehyde levels. In addition, compared with CT group, intestinal digestive ability was substantially enhanced as indicated by the significantly increased lipase activity in BA2 group (P < 0.05) and significantly increased amylase activity in BA1 and BA2 groups (P < 0.05). Coincidentally, BAs inclusion significantly upregulated the relative expression of intestinal mucosal barrier-related genes (P < 0.05). Further, dietary BAs distinctly remodeled intestinal microbiota by decreased the abundance of some potential pathogenic bacteria. In conclusion, dietary BAs supplementation is an effective way to improve the intestinal healthy status of tongue sole.

Mechanism of persistence of indigenous bifidobacteria under the impact of acetate in the human colon biotope

Aim. To determine the role of the acetate in the persistence of indigenous bifidobacteria in the colon biotope through the lysozyme resistance in model conditions of the acetylation–deacetylation of peptidoglycan.Materials and methods. The study was performed on 16 strains of the two indigenous bifidobacteria speсies: Bifidobacterium bifidum и Bifidobacterium longum subsp. longum. Bifidobacteria was cultivated in the 0.6% O2 and 9% CO2 atmosphere at the temperature 37ºС in CO2 incubator for 48 hours. The production of the acetate by the bifidobacteria was determined by gas chromatography. The effect of the acetate on the lysozyme resistance of non-indigenous gram-positive bacteria was determined on the Listeria monocytogenes ICIS-280 model strain by the cultivation in LB-Lennox broth with ammonium acetate added in the concentration range matching the concentrations produced by the studied bifidobacteria, in lysozyme serial dilutions at final concentrations 5 μg/ml to 40 μg/ml within 24 hours.Results. It was found that the acetate release of Bifidobacterium longum subsp. longum was on average two times higher that of Bifidobacterium bifidum (27.0 and 14.7 mmol/liter, respectively) and was quite consistent with the concentrations of acetic acid determined in the intestinal contents (up to 50 mmol/liter). Cultivation of bifidobacteria in a medium with lysozyme, ammonium acetate and their combination did not have a significant impact on their growth parameters at the maximum used concentrations of these substances. In the test strain, the addition of ammonium acetate in the range created by bifidobacteria caused a decrease in the minimum inhibitory concentration of lysozyme by more than two times — from 40 μg/ml to less than 20 μg/ml. In the control medium without lysozyme, no inhibition of the growth of the indicator culture was observed up to the maximum concentrations of ammonium acetate.Conclusion. The mechanism of persistence (survival) of indigenous bifidobacteria in the human intestinal biotope has been identified, which is associated with the production of acetic acid at a level that selectively suppresses lysozyme resistance of non-indigenous gram-positive microbiota viareversible deacetylation of peptidoglycan. This allows indigenous bifidobacteria to maintain a stable dominant position in the biotope.

Influence of the Immunomodulator AVIGEN to Broiler Chicken Humoral Immune Factors

The nonspecific immune response plays an important role in organism's defense against a variety of pathogens. Two major factors in this process are blood serum lysozyme and alternative pathway of complement activation (APCA). Over the past few decades, the application of various substances, targeting improved levels of natural immunity have become part of the mainstream trends in livestock rearing programs. The current study examined the influence of the polybacterial immunomodulator AVIGEN on the performance of the aforementioned immune factors among broiler chicken hybrids. The experimental group demonstrated better overall performance for both parameters of interest. APCA activity for the treated group (549.10±19.69 CH50) was significantly higher compared to the result obtained for the controls (377.40±9.58 CH50), (P<0.001). Results for the other parameter were even more indicative. The measured concentration of the serum lysozyme for the treated group was twice as high compared to the control birds - 6.17±0.49 mg/L vs. 2.99±0.27 mg/L, respectively. Therefore, the introduction of the AVIGEN immunomodulator to animals' diet has strong potential to improve natural humoral immunity in poultry farming.

Dietary Laminaria japonica Polysaccharide Regulates Systemic Defence Property of Weaned Piglets Improving Growth Performance under High Temperature Condition

Background: This study was conducted to evaluate the effect of dietary LJP supplementation on growth performance, heat stress resistance and immune status of weaned piglets raised in a high-temperature environment (34 ± 3°C).Methods: A total of 72 Barkshire × Licha Black crossbred piglets were weaned at 30 days of age (BW, 5.88 ± 0.16 kg) and fed either without or with 100, 200 or 400 mg/kg LJP in a completely randomized design, with 3 pens per diet (6 pigs per pen). On day 21, a total of 12 piglets were selected to collect blood samples.Result: The results showed that 200 and 400 mg/kg LJP supplementation decreased the feed conversion ratio (FCR) of piglets compared to the control group (P = 0.037) and dietary LJP linearly increased average daily gain and decreased FCR of piglets (P = 0.037 and 0.011, respectively). In addition, adding 100, 200 and 400 mg/kg LJP reduced the serum HSP70 content compared to the control group (P = 0.019) and dietary LJP linearly and quadratically decreased the serum HSP70 content of piglets (P = 0.030 and 0.011, respectively). Moreover, supplementation of 200 mg/kg and 400 mg/kg LJP increased the serum (NO) and nitric oxide acid phosphatase (ACP) content compared to the control group (P = 0.019; P = 0.003) and dietary LJP linearly increased the serum lysozyme, NO and ACP contents (P = 0.039; P = 0.005; P = 0.001). In conclusion, dietary LJP has potential to regulate systemic defence property of weaned piglets raised in a high temperature condition, accompanied by increased growth performance.

Global assessment of the response to chronic stress in European sea bass

Stress modifies energy allocation in fishes by redirecting energy from growth and reproduction to coping mechanisms. However, these adjustments become inappropriate when the challenge consists of sustained or repeated stressors, with the animal entering a maladaptive state. Capacities to cope with additional threats are then altered and compromise survival. The characterization of the responses to chronic stress in fishes helps better understanding the physiological limits in an aquaculture or ecological context. Here, we investigate the coping capacities of European sea bass to multiple and diverse stressors applied over a 3-weeks period. Multiple behavioural (group dispersion and swimming activity) and physiological responses (blood cortisol, osmoregulatory mechanisms, stress-related gene expression, etc.) were evaluated in resting fish or in fish exposed to additional challenges. Resilience to the chronic stress protocol was evaluated 4 months after the end of the chronic stress. Chronically stressed individuals showed reduced growth, lower cortisol response, increased chloride and sodium concentration in the plasma and modified gill gene expression translating osmoregulatory dysfunctions. Chronic stress had no significant effect on plasmatic calcium, lysozyme concentration and osmotic pressure. Increased thigmotaxic behaviour was observed in a new environment behavioural test. Four months after the chronic stress, no significant difference was observed in growth performances and in plasma parameters. Altogether, gills and more generally osmoregulatory functions were found to be the most sensitive to the chronic stress, while only limited changes in growth, activity of the HPI axis, immunity and swimming behaviour were observed when assessed individually. This work demonstrates the necessity of using multiple and diverse endpoints related to different functions to properly assess health and welfare in fishes.