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BMC Veterinary Research Jan 2024Attempts to use dietary lysozyme (LYZ) as an alternative to antibiotics in broilers have been successful, but further research is needed for effective use. Here, we...
BACKGROUND
Attempts to use dietary lysozyme (LYZ) as an alternative to antibiotics in broilers have been successful, but further research is needed for effective use. Here, we compared the differences between LYZ and avilamycin (AVI) feed additives for growth performance, gut health and immunity of broilers. One-day old, one hundred and twenty broiler chicks (Ross 308) were randomly allocated into three groups consisting forty birds in each group. Standard diet without supplementation was applied as the control group (I), while the chicks of the other groups were supplemented with 100 mg of AVI per kg diet (AVI, group II), and 90 mg LYZ per kg diet (LYZ, group III) for five consecutive weeks.
RESULTS
Body weight, feed conversion ratio, body weight gain, and European production efficiency factor were markedly (p < 0.05) increased in both AVI and LYZ groups in relation to CON group, but the feed intake and protein efficiency ratio were not affected. Both AVI and LYZ significantly (p < 0.001) upregulated the mRNA expression of ileal interleukin-18 (IL-18), interferon-gamma (IFN-γ), and interleukin-10 (IL-10), interleukin-2 (IL-2), and glutathione peroxidase (GSH-PX) genes compared to CON group. However, IL-2, IL-10, IL-18, and GSH-PX genes were markedly (p < 0.01) upregulated in LYZ compared to the AVI group. LYZ treated group had a significant increase (p < 0.05) in the serological haemagglutination inhibition titers of H5N1 vaccination and a significant decrease (p < 0.0001) in coliform counts compared to control and AVI groups, but all growth parameters were nearly similar between AVI and LYZ groups. The VH and VH/CD were markedly higher in LYZ than AVI and control groups.
CONCLUSION
Exogenous dietary lysozyme supplementation by a dose of 90 mg/kg broilers' diet induced better effects on intestinal integrity, fecal bacterial counts, immune response, and growth performance which were comparable to avilamycin. Therefore, dietary lysozyme could safely replace avilamycin in the broiler chickens' diet. However, further experimental studies regarding the use of lysozyme in commercial broilers, both in vitro and in vivo, targeting more communities of intestinal microbiome and explaining more details about its beneficial effects need to be conducted.
Topics: Animals; Chickens; Interleukin-2; Interleukin-10; Interleukin-18; Muramidase; Influenza A Virus, H5N1 Subtype; Diet; Dietary Supplements; Body Weight; Animal Feed; Oligosaccharides
PubMed: 38245745
DOI: 10.1186/s12917-023-03871-2 -
Biomacromolecules Feb 2024The ability to fine-tune the volume phase transition temperature (VPTT) of thermoresponsive nanoparticles is essential to their successful application in drug delivery....
The ability to fine-tune the volume phase transition temperature (VPTT) of thermoresponsive nanoparticles is essential to their successful application in drug delivery. The rational design of these materials is limited by our understanding of the impact that nanoparticle-protein interactions have on their thermoresponsive behavior. In this work, we demonstrate how the formation of protein corona impacts the transition temperature values of acrylamide-based nanogels and their reversibility characteristics, in the presence of lysozyme, given its relevance for the ocular and intranasal administration route. Nanogels were synthesized with -isopropylacrylamide or --propylacrylamide as backbone monomers, methylenebis(acrylamide) (2.5-20 molar %) as a cross-linker, and functionalized with negatively charged monomers 2-acrylamido-2-methylpropanesulfonic acid, -acryloyl-l-proline, or acrylic acid; characterization showed comparable particle diameter (.10 nm), but formulation-dependent thermoresponsive properties, in the range 28-54 °C. Lysozyme was shown to form a complex with the negatively charged nanogels, lowering their VPTT values; the hydrophilic nature of the charged comonomer controlled the drop in VPTT upon complex formation, while matrix rigidity only had a small, yet significant effect. The cross-linker content was found to play a major role in determining the reversibility of the temperature-dependent transition of the complexes, with only 20 molar % cross-linked-nanogels displaying a fully reversible transition. These results demonstrate the importance of evaluating protein corona formation in the development of drug delivery systems based on thermoresponsive nanoparticles.
Topics: Nanogels; Protein Corona; Muramidase; Acrylamide; Drug Carriers; Temperature; Acrylamides
PubMed: 38242644
DOI: 10.1021/acs.biomac.3c01405 -
Molecules (Basel, Switzerland) Dec 2023Optical methods (spectroscopy, spectrofluorometry, dynamic light scattering, and refractometry) were used to study the change in the state of hen egg-white lysozyme...
Optical methods (spectroscopy, spectrofluorometry, dynamic light scattering, and refractometry) were used to study the change in the state of hen egg-white lysozyme (HEWL), protein molecules, and gold nanoparticles (AuNPs) in aqueous colloids with changes in pH, and the interaction of protein molecules with nanoparticles was also studied. It was shown that changing pH may be the easiest way to control the protein corona on gold nanoparticles. In a colloid of nanoparticles, both in the presence and absence of protein, aggregation-deaggregation, and in a protein colloid, monomerization-dimerization-aggregation are the main processes when pH is changed. A specific point at pH 7.5, where a transition of the colloidal system from one state to another is observed, has been found using all the optical methods mentioned. It has been shown that gold nanoparticles can stabilize HEWL protein molecules at alkaline pH while maintaining enzymatic activity, which can be used in practice. The data obtained in this manuscript allow for the state of HEWL colloids and gold nanoparticles to be monitored using one or two simple and accessible optical methods.
Topics: Muramidase; Gold; Metal Nanoparticles; Colloids; Hydrogen-Ion Concentration
PubMed: 38202662
DOI: 10.3390/molecules29010082 -
BMC Veterinary Research Jan 2024The mare-foal relationship is essential for the well-being and growth of a foal. Mare's milk provides a foal with nutrients, protective immunity, and microbes. Within...
BACKGROUND
The mare-foal relationship is essential for the well-being and growth of a foal. Mare's milk provides a foal with nutrients, protective immunity, and microbes. Within the first two weeks of life, there is a risk for a foal to suffer from diarrhea, particularly "foal heat diarrhea" which happens at about the time of a mare's estrus cycle but is more likely due to transitions in the microbiota in the foal's gastrointestinal (GI) tract. We hypothesized that this GI microbiota transition could be caused by changes in lysozyme and microbial populations in the mare's milk. To test this hypothesis, fifteen mare-foal pairs were followed in the first 15 days post-foaling. Every other day milk was collected from mares and rectal swabs were collected from foals. Lysozyme activity in the mare's milk was measured using a fluorescence assay. Microbial DNA was isolated from the milk and swabs and the V4 domain of 16 S rRNA genes were PCR amplified and sequenced using Illumina MiSeq technology. Microbial populations were analyzed using DADA2 and phyloseq within R.
RESULTS
Mare's milk lysozyme activity peaked for samples at Day 1 and levels dropped to 72.5% of Day 1 activity by Day 15; however, microbial populations in the mare's milk did not vary significantly over the two weeks. Furthermore, levels of microbial diversity found in foal rectal swabs were initially similar to microbial diversity seen in mare's milk; however, over the first fifteen days, diversity increased for the foal rectal swab microbiota and swab microbial populations differed from milk microbes. A transition occurred shifting from microbes from the phylum Proteobacteria early in rectal swabs to those primarily from the phyla Firmicutes and Bacteroidota after the first few days post-foaling. These phyla contained several families and genera of microbes that promote utilization of milk components in healthy gut transition. Microbial abundance levels correlated more with days post-parturition than with lysozyme activity and mare's milk microbial populations.
CONCLUSIONS
The findings suggest that much of the microbial populations responsible for the transition of the foal's gut comes from sources outside of mare's milk species and levels of lysozyme activity.
Topics: Humans; Animals; Female; Horses; Milk; Muramidase; Microbiota; Gastrointestinal Microbiome; Diarrhea
PubMed: 38191395
DOI: 10.1186/s12917-023-03864-1 -
International Journal of Molecular... Dec 2023Verticillium wilt is a soil-borne vascular disease caused by the fungal pathogen . It causes great harm to upland cotton () yield and quality. A previous study has shown...
Verticillium wilt is a soil-borne vascular disease caused by the fungal pathogen . It causes great harm to upland cotton () yield and quality. A previous study has shown that Hen egg white lysozyme (HEWL) exerts strong inhibitory activity against in vitro. In the current study, we introduced the gene into cotton through the -mediated transformation, and the exogenous HEWL protein was successfully expressed in cotton. Our study revealed that HEWL was able to significantly inhibit the proliferation of in cotton. Consequently, the overexpression of HEWL effectively improved the resistance to Verticillium wilt in transgenic cotton. In addition, ROS accumulation and NO content increased rapidly after the inoculation of plant leaves overexpressing HEWL. In addition, the expression of the PR genes was significantly up-regulated. Taken together, our results suggest that HEWL significantly improves resistance to Verticillium wilt by inhibiting the growth of pathogenic fungus, triggering ROS burst, and activating PR genes expression in cotton.
Topics: Gossypium; Reactive Oxygen Species; Verticillium; Muramidase; Egg White; Disease Resistance; Plant Diseases; Gene Expression Regulation, Plant; Plant Proteins
PubMed: 38138993
DOI: 10.3390/ijms242417164 -
Microorganisms Dec 2023The ability of epithelial barriers to perform as the first defense line against external damage derives from tight junctions, protein complexes that block microorganisms...
The ability of epithelial barriers to perform as the first defense line against external damage derives from tight junctions, protein complexes that block microorganisms through the paracellular space. Indeed, disturbances of barrier permeability caused by bacterial metabolites and other inflammatory stimuli are the consequence of changes in protein expression in these complexes. Postbiotics, molecules derived from bacteria with beneficial effects on the host, improve barrier function through the activation of survival pathways in epithelial cells. GG secretes the muramidase p40, which protects intestinal barriers through an EGFR-dependent pathway. In this work, we cloned, expressed, and purified the recombinant p40 protein from GR-1 to evaluate its effect on cell viability, cell cytotoxicity, TEER, and protein levels of tight junctions, as well as EGFR activation via Western blot on HaCaT keratinocytes subjected to LPS. We found a novel mutation at residue 368 that does not change the structure of p40. Our protein also reduces the LPS-induced increase in cell cytotoxicity when it is added prior to this stimulus. Furthermore, although LPS did not cause changes in barrier function, p40 increased TEER and occludin expression in HaCaT, but unlike previous work with p40 from LGG, we found that recombinant p40 did not activate EGFR. This suggests that recombinant p40 enhances epithelial barrier function through distinct signaling pathways.
PubMed: 38138057
DOI: 10.3390/microorganisms11122913 -
Animals : An Open Access Journal From... Dec 2023The impact of microbial muramidase (MMUR) addition to broiler chicken rations was evaluated through growth parameters, liver histoarchitecture, antioxidant status,...
Effects of Dietary Microbial Muramidase on the Growth, Liver Histoarchitecture, Antioxidant Status, and Immunoexpression of Pro-Inflammatory Cytokines in Broiler Chickens.
The impact of microbial muramidase (MMUR) addition to broiler chicken rations was evaluated through growth parameters, liver histoarchitecture, antioxidant status, biochemical analysis, and expression of pro-inflammatory cytokines for 35 days. Four hundred three-day-old chicks (97.68 ± 0.59 g) were distributed to four distinct groups with ten duplicates each (100 chicks/group) consisting of: group 1 (G1): a basal diet without MMUR (control group); G2: a basal diet + 200 mg MMUR kg G3: a basal diet + 400 mg MMUR kg; and G4: a basal diet + 600 mg MMUR kg. The results showed that the final body weight and total weight gain were increased ( = 0.015) in birds fed with diets supplemented with MMUR at 600 mg kg. The feed conversion ratio (FCR) was improved in all treatment groups compared with the control group. Birds fed with a diet supplemented with 600 mg MMUR kg showed the highest body weight gain and improved FCR. The values of thyroxin hormones and growth hormones were increased in all MMUR-supplemented groups. Dietary MMUR increased the activities of antioxidant enzymes (total antioxidant activity, catalase, and superoxide dismutase) and decreased the activity of malondialdehyde ( < 0.05). In addition, it increased the values of interleukin 1 beta and interferon-gamma compared with the control group. Furthermore, dietary MMUR increased the expression of transforming growth factor-beta immunostaining in the liver and spleen tissues. Our results show that supplementing broilers' diets with 600 mg MMUR kg could enhance the chicken growth rate and improve their antioxidant, inflammatory, and anti-inflammatory responses.
PubMed: 38136899
DOI: 10.3390/ani13243862 -
Poultry Science Feb 2024The study aimed to analyze the biological value of eggs and extra-embryonic structures affecting pheasant hatchability depending on the eggshell's color. Eggs (1,415)...
The study aimed to analyze the biological value of eggs and extra-embryonic structures affecting pheasant hatchability depending on the eggshell's color. Eggs (1,415) from 62-wk-old pheasants were used. The quality of fresh blue (BL), brown (BR), and green (G) eggs were analyzed. Incubation lasted for 25 d. Thick albumen (d 0, 1, 7, 14), amniotic fluid (d 14, 18), and the yolk (d 0-14) were collected. The pH, viscosity, lysozyme activity, crude protein (CP) content in albumen and amnion, pH, vitelline membrane strength, and fatty acids (FA) content in the yolk were performed. The lowest hatchability was in the BL group, and the highest was in the G group. BL group showed lower eggshell thickness and strength and higher egg weight. In thick albumen and amniotic fluid, the pH decreased with the incubation. In the yolk, there was an increasing trend (P = 0.015), with a decrease on d 18 (P < 0.001). The vitelline membrane strength decreased after 1 d of incubation, excluding BR eggs (P < 0.001). Thick albumen viscosity was higher on d 14 in the G group than in other dates and groups, the lowest in amniotic fluid, and slightly higher in BL and BR eggs. On d 18, amniotic fluid viscosity increased (P < 0.001). The lowest viscosity was indicated in BL eggs (P < 0.001). The lysozyme activity in thick albumen on d 14 was the highest (uniquely in BR and G groups), and the lowest values were found in amniotic fluid on d 14; after four d, the activity increased (P < 0.001). The CP content was higher in the BL group on d 14. In amnion, on d 14, the CP content was the lowest (<1%) and increased on d 18 (P < 0.001). There was a higher FA content (especially UFA) in the G group and a decrease in FA content after d 14 (P < 0.001). It was found that eggs with green eggshells have the highest biological value, and blue eggs are the least useful for incubation.
Topics: Animals; Chickens; Egg Shell; Muramidase; Ovum; Meat; Albumins; Quail; Fatty Acids; Eggs; Egg Yolk
PubMed: 38134460
DOI: 10.1016/j.psj.2023.103338 -
Biosensors Nov 2023Nonspecific adsorption has always been a critical challenge for sensor detection; thus, an efficient and facile approach for fabricating antifouling sensors is highly...
Nonspecific adsorption has always been a critical challenge for sensor detection; thus, an efficient and facile approach for fabricating antifouling sensors is highly desirable. Here, we developed an antifouling coating on sensor surfaces, conveniently made with a simple drip of phase-transited BSA (PTB) followed by a modification with a peanut allergen antibody, which unexpectedly provides synergistic antifouling properties in sensors. Atomic force microscopy and scanning electron microscopy were used to evaluate the surface evenness. Optimizations in terms of PTB modification time and concentrations were performed using surface plasmon resonance by measuring protein resistance capabilities. Compared to bare Au surfaces, the PTB-modified surfaces exhibited low adsorption against BSA (<10 ng/cm) and good resistance against lysozyme (Lyz). After immobilizing antibodies, the antifouling performance of the sensor coatings had an obvious enhancement, with almost no BSA adsorption and low lysozyme adsorption. The target recognition was also analyzed to verify the good sensing performance of the antifouling sensor. This understanding of antibody synergy provides suggestions for the development of antifouling sensors.
Topics: Muramidase; Biofouling; Antibodies; Proteins; Surface Plasmon Resonance; Surface Properties
PubMed: 38131764
DOI: 10.3390/bios13121004 -
International Journal of Molecular... Nov 2023Cows produce saliva in very large quantities to lubricate and facilitate food processing. Estimates indicate an amount of 50-150 L per day. Human saliva has previously...
Cows produce saliva in very large quantities to lubricate and facilitate food processing. Estimates indicate an amount of 50-150 L per day. Human saliva has previously been found to contain numerous antibacterial components, such as lysozyme, histatins, members of the S-100 family and lactoferrin, to limit pathogen colonization. Cows depend on a complex microbial community in their digestive system for food digestion. Our aim here was to analyze how this would influence the content of their saliva. We therefore sampled saliva from five humans and both nose secretions and saliva from six cows and separated the saliva on SDS-PAGE gradient gels and analyzed the major protein bands with LC-MS/MS. The cow saliva was found to be dominated by a few major proteins only, carbonic anhydrase 6, a pH-stabilizing enzyme and the short palate, lung and nasal epithelium carcinoma-associated protein 2A (SPLUNC2A), also named bovine salivary protein 30 kDa (BSP30) or BPIFA2B. This latter protein has been proposed to play a role in local antibacterial response by binding bacterial lipopolysaccharides (LPSs) and inhibiting bacterial growth but may instead, according to more recent data, primarily have surfactant activity. Numerous peptide fragments of mucin-5B were also detected in different regions of the gel in the MS analysis. Interestingly, no major band on gel was detected representing any of the antibacterial proteins, indicating that cows may produce them at very low levels that do not harm the microbial flora of their digestive system. The nose secretions of the cows primarily contained the odorant protein, a protein thought to be involved in enhancing the sense of smell of the olfactory receptors and the possibility of quickly sensing potential poisonous food components. High levels of secretory IgA were also found in one sample of cow mouth drippings, indicating a strong upregulation during an infection. The human saliva was more complex, containing secretory IgA, amylase, carbonic anhydrase 6, lysozyme, histatins and a number of other less abundant proteins, indicating a major difference to the saliva of cows that show very low levels of antibacterial components, most likely to not harm the microbial flora of the rumen.
Topics: Humans; Female; Cattle; Animals; Saliva; Muramidase; Histatins; Chromatography, Liquid; Tandem Mass Spectrometry; Salivary Proteins and Peptides; Immunoglobulin A, Secretory; Anti-Bacterial Agents
PubMed: 38069163
DOI: 10.3390/ijms242316838