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Advances in Colloid and Interface... Oct 2019The design of immobilized enzyme preparations is an important and relevant area of modern sciences and technologies. Immobilization of enzymes from animal sources... (Review)
Review
The design of immobilized enzyme preparations is an important and relevant area of modern sciences and technologies. Immobilization of enzymes from animal sources (component I) on natural carriers (component II) increases the system stability by protecting the active site of the enzyme from deactivation; facilitates the separation and accelerates the recovery of the enzyme. This makes reuse possible and provides a significant reduction in operating costs. Hydrolytic enzymes (such as lipases) and polysaccharides (such as chitosan) are the most promising of such pairs of components. The main attention here is devoted to the discussion on lipase immobilization on polysaccharide (mainly - chitin and chitosan). Based on the analysis of the available literature, the most adequate method is the immobilization of lipase from porcine pancreas (LPP) on polysaccharide particles (such as chitin or chitosan) pre-treated with ultrasound (to increase the particle surface area) and glutaraldehyde (for particle activation) that shows reasonably high LPP activity and stability. In order to increase further the activity of the lipase, some authors proposed to incorporate a spacer in the form of 1,3-diaminopropane (or 1,3-diaminobutane) prior to activation of the surface of the chitosan particles. In particular cases, the use of chitin (instead of chitosan) may be an alternative solution for biotechnological applications. Recently the idea of constructing "supramolecular enzyme systems" realized in the so-called "coimmobilized multienzymatic systems" strategy. The most fascinating example is the combined assay of a mixture of native LPP, glycerol kinase (from Cellulomonas) and glycerol-3-phosphate oxidase (from Aerococcus viridans) linked by glutaraldehyde to chitosan (as shell for inorganic nanoparticle core). This material was placed on a Pt-electrode as biosensor and was successfully applied for amperometric determination of the triglyceride level in the serum of healthy and diseased person. Thus, the whole innovative research-production sequence is described by Aggarwal V. and Pundir C.S.: from simple components to advanced material and further biomedical application. Thus, the following approach of lipase immobilization appears the most promising for future applications: a few types of lipases or the combination of LPP with some other enzymes immobilized simultaneously on multifunctional carriers (as nanohybrids of inorganic core and polysaccharide shell).
Topics: Animals; Biotechnology; Chitin; Chitosan; Enzymes, Immobilized; Glutaral; Lipase; Nanoparticles; Polysaccharides; Swine
PubMed: 31421454
DOI: 10.1016/j.cis.2019.102016 -
Journal of Medical Microbiology Jul 2021Periodontitis, one of the most common oral disorders in sheep, is caused by a mixed and opportunistic microbiota that severely affects the health and welfare of...
Periodontitis, one of the most common oral disorders in sheep, is caused by a mixed and opportunistic microbiota that severely affects the health and welfare of animals. However, little is known about the ecological processes involved and the composition of the microbiota associated with the development of the disease. Using high-throughput sequencing of the 16S ribosomal RNA gene and network analysis it would be possible to discriminate the microbiomes of clinically healthy sheep and those with periodontitis and possibly identify the key microorganisms associated with the disease. The present study aimed to characterise the composition of dental microbiomes and bacterial co-occurrence networks in clinically healthy sheep and animals with periodontitis. Dental biofilm samples were collected from ten sheep with periodontitis and ten clinically healthy animals. Bacteria were identified using high-throughput sequencing of the 16S ribosomal RNA gene. The most prevalent genera in the dental microbiota of sheep with periodontitis were , , and . In clinically healthy animals, the most significant genera were unclassified and was found at high prevalence in the microbiomes of both groups. The dental microbiota of sheep in the two clinical conditions presented different profiles and the diversity and richness of bacteria was greater in the diseased animals. Network analyses showed the presence of a large number of antagonistic interactions between bacteria in the dental microbiota of animals with periodontitis, indicating the occurrence of a dysbiotic community. Through the interrelationships, members of the genus are likely to be key pathogens, both in the dental microbiota of healthy animals and those with periodontitis. stood out among the top three nodes with more centrality and the largest number of hubs in the networks of animals with periodontitis. The dental biofilm microbiota associated with ovine periodontitis is dysbiotic and with significant antagonistic interactions, which discriminates healthy animals from diseased animals and highlights the importance of key bacteria, such as , , and species.
Topics: Animals; Bacteria; Biofilms; Ecology; High-Throughput Nucleotide Sequencing; Microbiota; Periodontitis; RNA, Ribosomal, 16S; Sheep
PubMed: 34313584
DOI: 10.1099/jmm.0.001396 -
BMC Microbiology Oct 2020Antibiotic-associated diarrhea (AAD), defined as diarrhea that occurs in association with the administration of antibiotics and without another clear etiology, is one of...
BACKGROUND
Antibiotic-associated diarrhea (AAD), defined as diarrhea that occurs in association with the administration of antibiotics and without another clear etiology, is one of the most common adverse drug events of antibiotics therapy. We established a diarrhea model induced by gentamycin and cefradine to investigate the microbiota characteristics in the intestinal lumen of mice with AAD and provide insights into noteworthy bacteria related to gentamicin and cefradine-associated diarrhea.
RESULTS
The number of OTUs in the model group and the normal group was 983 and 2107, respectively, and 872 identical OTUs were shared between two groups. Species richness and species diversity of intestinal microbe were altered by antibiotics administration. PCoA showed a clear separation between AAD and health control. The dominant phyla of AAD mice were Firmicutes (52.63%) and Proteobacteria (46.37%). Among the genus with top 20 abundance, the relative abundance of 7 genera, Ruminococcus, Blautia, Enterococcus, Eubacterium, Clostridium, Coprococcus, and Aerococcus, were enriched in the model group. Based upon the LEfSe analysis, Enterococcus, Eubacterium, Ruminococcus, and Blautia were identified as potential biomarkers for AAD.
CONCLUSIONS
The bacterial diversity of the intestinal lumen was diminished after gentamicin and cefradine administration. The alterations in the abundance and composition of gut microbiota further led to the dysfunction of gut microbiota. More specifically, gentamicin and cefradine significantly increased the abundance of the opportunistic pathogens, of which Enterococcus and Clostridium were the most prominent and most worthy of attention.
Topics: Animals; Anti-Bacterial Agents; Bacteria; Diarrhea; Female; Gastrointestinal Microbiome; Male; Mice; RNA, Ribosomal, 16S
PubMed: 33059603
DOI: 10.1186/s12866-020-01999-x -
Frontiers in Cellular and Infection... 2022Human papillomavirus (HPV) infection is one of the most common sexually transmitted diseases. After studying 602 unvaccinated Chinese women using 16S rRNA to detect...
Human papillomavirus (HPV) infection is one of the most common sexually transmitted diseases. After studying 602 unvaccinated Chinese women using 16S rRNA to detect cervical-vaginal microecology, we analyzed the relationship between HPV infection and vaginal microecology including 20 HPV types. In Chinese women, dominated and dominated clusters were significantly absence. Microbial alpha diversity was significantly higher in HPV-infected and cervical intraepithelial neoplasia (CIN)-diagnosed groups than in healthy control group. Certain bacteria were associated with HPV infection and CIN, including , , , , , and With the development of disease, the proportions of community state type III (CST-III) and CST-IV-B gradually increased, whereas the proportions of CST-I and CST-IV-A gradually decreased. In addition, age was an influential factor for HPV infection. With aging, the probability of HPV infection and the proportion of CST-IV-B increase. In conclusion, our study was a large cross-sectional study that evaluated the relationship between vaginal microbiota and HPV infection, and brought essential comparable data.
Topics: China; Cross-Sectional Studies; Female; Humans; Lactobacillus; Microbiota; Papillomavirus Infections; RNA, Ribosomal, 16S; Vagina
PubMed: 35800384
DOI: 10.3389/fcimb.2022.814668 -
International Journal of Systematic and... Nov 2023A non-spore-forming, Gram-stain-positive, short rod-shaped strain, designated SJQ22, was isolated from a paddy soil sample collected in Shanghai, PR China. A comparative...
A non-spore-forming, Gram-stain-positive, short rod-shaped strain, designated SJQ22, was isolated from a paddy soil sample collected in Shanghai, PR China. A comparative analysis of 16S rRNA gene sequences showed that strain SJQ22 fell within the genus , forming a clear cluster with the type strains of (98.6 % sequence similarity) and (98.5 % sequence similarity). Strain SJQ22 grew at 30-45 °C (optimum, 30 °C), pH 6.0-8.0 (optimum, pH 7.0) and with a NaCl concentration of 0-4 % (optimum, 1 %). Cells were negative for oxidase and catalase activity. Chemotaxonomic analysis showed that strain SJQ22 possessed C and C ω9 as the predominant fatty acids. The DNA G + C content was 39.0 mol%. Strain SJQ22 exhibited DNA-DNA relatedness levels of 13±2 % with ATCC 11563 and 9±2 % with IFO 12173. Based on the data obtained, strain SJQ22 represents a novel species of the genus , for which the name sp. nov. is proposed. The type strain is SJQ22 (=JCM 33111=CCTCC AB 2018283).
Topics: Fatty Acids; Soil Microbiology; Aerococcus; RNA, Ribosomal, 16S; DNA, Bacterial; Nucleic Acid Hybridization; Base Composition; China; Phylogeny; Bacterial Typing Techniques; Sequence Analysis, DNA
PubMed: 37909297
DOI: 10.1099/ijsem.0.006069 -
Microorganisms Nov 2021The investigation of bacterial microbiota represents a developing research field in veterinary medicine intended to look for correlations between animal health and the...
The investigation of bacterial microbiota represents a developing research field in veterinary medicine intended to look for correlations between animal health and the balance within bacterial populations. The aim of the present work was to define the bacterial microbiota of the oral cavity of healthy sows, which had not been thoroughly described so far. In total, 22 samples of oral fluid were collected and analyzed by 16S-rRNA gene sequencing. CLC Genomics Workbench 20.0 (QIAGEN Digital Insights, Aarhus, Denmark) was then used to examine the results. The predominant orders were Lactobacillales, Clostridiales, and Corynebacteriales. Lactobacillaceae, Corynebacteriaceae, Moraxellaceae, Aerococcaceae, and Staphylococcaceae were the most represented families. As regards the most abundant genera, , , , , , , and can be pointed out as the bacterial core microbiota. Sows were also divided into "gestating" and "lactating" groups, and mild differences were found between pregnant and lactating sows. The data herein described represent an original contribution to the knowledge of the porcine bacterial microbiota. Moreover, the choice of sows as experimental animals was strategic for identifying the adult microbial community. These data provide a basis for further studies on the oral bacterial microbiota of pigs.
PubMed: 34835441
DOI: 10.3390/microorganisms9112314 -
Journal of Biotechnology Feb 2024l-Lactate oxidase has important applications in biosensing and finds increased use in biocatalysis. The enzyme has been characterized well, yet its immobilization has...
l-Lactate oxidase has important applications in biosensing and finds increased use in biocatalysis. The enzyme has been characterized well, yet its immobilization has not been explored in depth. Here, we studied immobilization of Aerococcus viridansl-lactate oxidase on porous carriers of variable matrix material (polymethacrylate, polyurethane, agarose) and surface functional group (amine, Ni-loaded nitrilotriacetic acid (NiNTA), epoxide). Carrier activity (A) and immobilized enzyme effectiveness (ɳ) were evaluated in dependence of protein loading. Results show that efficient immobilization (A: up to 1450 U/g carrier; ɳ: up to 65%) requires a hydrophilic carrier (agarose) equipped with amine groups. The value of ɳ declines sharply as A increases, probably due to transition into diffusional regime. Untagged l-lactate oxidase binds to NiNTA carrier similarly as N-terminally His-tagged enzyme. Lixiviation studies reveal quasi-irreversible enzyme adsorption on NiNTA carrier while partial release of activity (≤ 25%) is shown from amine carrier. The desorbed enzyme exhibits the same specific activity as the original l-lactate oxidase. Collectively, our study identifies basic requirements of l-lactate oxidase immobilization on solid carrier and highlights the role of ionic interactions in enzyme-surface adsorption.
Topics: Aerococcus; Sepharose; Mixed Function Oxygenases; Enzymes, Immobilized; Amines
PubMed: 38280467
DOI: 10.1016/j.jbiotec.2024.01.011 -
Clinical Chemistry and Laboratory... Jun 2024The EFLM Task and Finish Group Urinalysis has updated the ECLM European Urinalysis Guidelines (2000) on urinalysis and urine bacterial culture, to improve accuracy of...
BACKGROUND
The EFLM Task and Finish Group Urinalysis has updated the ECLM European Urinalysis Guidelines (2000) on urinalysis and urine bacterial culture, to improve accuracy of these examinations in European clinical laboratories, and to support diagnostic industry to develop new technologies.
RECOMMENDATIONS
Graded recommendations were built in the following areas.
MEDICAL NEEDS AND TEST REQUISITION
Strategies of urine testing are described to patients with complicated or uncomplicated urinary tract infection (UTI), and high or low-risk to kidney disease.
SPECIMEN COLLECTION
Patient preparation, and urine collection are supported with two quality indicators: contamination rate (cultures), and density of urine (chemistry, particles).
CHEMISTRY
Measurements of both urine albumin and α1-microglobulin are recommended for sensitive detection of kidney disease in high-risk patients. Performance specifications are given for urine protein measurements and quality control of multiproperty strip tests.
PARTICLES
Procedures for microscopy are reviewed for diagnostic urine particles, including urine bacteria. Technologies in automated particle counting and visual microscopy are updated with advice how to verify new instruments with the reference microscopy.
BACTERIOLOGY
Chromogenic agar is recommended as primary medium in urine cultures. Limits of significant growth are reviewed, with an optimised workflow for routine specimens, using leukocyturia to reduce less important antimicrobial susceptibility testing. Automation in bacteriology is encouraged to shorten turn-around times. Matrix assisted laser desorption ionization time-of-flight mass spectrometry is applicable for rapid identification of uropathogens. and are taken into the list of uropathogens. A reference examination procedure was developed for urine bacterial cultures.
PubMed: 38534005
DOI: 10.1515/cclm-2024-0070 -
Critical Reviews in Food Science and... 2024The gut microbiota (GM) is a complex ecosystem that is closely linked to host health. spp. polysaccharides (GPs), a major bioactive component of the fungal genus , can... (Review)
Review
The gut microbiota (GM) is a complex ecosystem that is closely linked to host health. spp. polysaccharides (GPs), a major bioactive component of the fungal genus , can modulate the GM, exhibiting various health effects and prebiotic potential. This review comprehensively concluded the structural features and extraction method of GPs. The mechanism of GPs for anti-obesity, anti-diabetes, anti-inflammatory, and anti-cancer were further evaluated. The simulated gastrointestinal digestion of GPs and the utilization mechanism of host microorganisms were discussed. It was found that the physicochemical properties and biological activities of GPs depend on their structural characteristics (molecular weight, monosaccharide composition, glycosidic bonds, etc.). Their extraction method also affects the structure and bioactivities of polysaccharides. GPs supplementation could increase the relative abundance of beneficial bacteria (e.g. , , and ), while reducing that of pathogenic bacteria (e.g. , ), thus promoting health. Moreover, GPs are resistant to digestion in the stomach and small intestine but are digested in the large intestine. Therefore, GPs can be considered as potential prebiotics. However, further studies should investigate how GPs as prebiotics regulate GM and improve host health.
Topics: Bacteria; Gastrointestinal Microbiome; Polysaccharides; Prebiotics
PubMed: 35980144
DOI: 10.1080/10408398.2022.2110035 -
BMC Microbiology Mar 2021Composition and maintenance of the microbiome is vital to gut homeostasis. However, there is limited knowledge regarding the impact of high doses of radiation, which can...
BACKGROUND
Composition and maintenance of the microbiome is vital to gut homeostasis. However, there is limited knowledge regarding the impact of high doses of radiation, which can occur as a result of cancer radiation therapy, nuclear accidents or intentional release of a nuclear or radioactive weapon, on the composition of the gut microbiome. Therefore, we sought to analyze alterations to the gut microbiome of nonhuman primates (NHPs) exposed to high doses of radiation. Fecal samples were collected from 19 NHPs (Chinese rhesus macaques, Macaca mulatta) 1 day prior and 1 and 4 days after exposure to 7.4 Gy cobalt-60 gamma-radiation (LD). The 16S V4 rRNA sequences were extracted from each sample, followed by bioinformatics analysis using the QIIME platform.
RESULTS
Alpha Diversity (Shannon Diversity Index), revealed no major difference between pre- and post-irradiation, whereas Beta diversity analysis showed significant differences in the microbiome after irradiation (day + 4) compared to baseline (pre-irradiation). The Firmicutes/Bacteriodetes ratio, a factor known to be associated with disruption of metabolic homeostasis, decreased from 1.2 to less than 1 post-radiation exposure. Actinobacillus, Bacteroides, Prevotella (Paraprevotellaceae family) and Veillonella genera were significantly increased by more than 2-fold and Acinetobacter and Aerococcus genus were decreased by more than 10-fold post-irradiation. Fifty-two percent (10/19) of animals exposed to radiation demonstrated diarrhea at day 4 post-irradiation. Comparison of microbiome composition of feces from animals with and without diarrhea at day 4 post-irradiation revealed an increase in Lactobacillus reuteri associated with diarrhea and a decrease of Lentisphaerae and Verrucomicrobioa phyla and Bacteroides in animals exhibiting diarrhea. Animals with diarrhea at day 4 post-irradiation, had significantly lower levels of Lentisphaere and Verrucomicrobia phyla and Bacteroides genus at baseline before irradiation, suggesting a potential association between the prevalence of microbiomes and differential susceptibility to radiation-induced diarrhea.
CONCLUSIONS
Our findings demonstrate that substantial alterations in the microbiome composition of NHPs occur following radiation injury and provide insight into early changes with high-dose, whole-body radiation exposure. Future studies will help identify microbiome biomarkers of radiation exposure and develop effective therapeutic intervention to mitigate the radiation injury.
Topics: Animals; Bacteria; Feces; Gamma Rays; Gastrointestinal Microbiome; Macaca mulatta; RNA, Ribosomal, 16S; Radiation Injuries
PubMed: 33781201
DOI: 10.1186/s12866-021-02146-w