Bioflocs are microbial aggregates primarily composed of heterotrophic bacteria that play essential ecological roles in maintaining animal health, gut microbiota, and water quality in biofloc aquaculture systems. Despite the global adoption of biofloc aquaculture for shrimp and fish cultivation, our understanding of biofloc microbiota-particularly the dominant bacterial members and their ecological functions-remains limited. In this study, we employed integrated metataxonomic and metagenomic approaches to demonstrate that the family Rhodobacteraceae of Alphaproteobacteria consistently dominates the biofloc microbiota and plays essential ecological roles. We first analyzed a comprehensive metataxonomic dataset consisting of 200 16S rRNA gene amplicons collected across three Asian countries: South Korea, China, and Vietnam.
Taxonomic investigation identified Rhodobacteraceae as the dominant and consistent bacterial members across the datasets. The predominance of this taxon was further validated through metagenomics approaches, including read taxonomy and read recruitment analyses. To explore the ecological roles of Rhodobacteraceae, we applied genome-centric metagenomics, reconstructing 45 metagenome-assembled genomes. Functional annotation of these genomes revealed that dominant Rhodobacteraceae genera, such as Marivita, Ruegeria, Dinoroseobacter, and Aliiroseovarius, are involved in vital ecological processes, including complex carbohydrate degradation, aerobic denitrification, assimilatory nitrate reduction, ammonium assimilation, and sulfur oxidation. Overall, our study reveals that the common practice of carbohydrate addition in biofloc aquaculture systems fosters the growth of specific heterotrophic bacterial communities, particularly Rhodobacteraceae. These bacteria contribute to maintaining water quality by removing toxic nitrogen and sulfur compounds and enhance animal health by colonizing gut microbiota and exerting probiotic effects.
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Two Gram-stain-positive, oxidase-negative, non-motile, facultative anaerobic, α-hemolytic, coccus-shaped bacteria (zg-86T and zg-70) were isolated from the respiratory tracts of marmots (Marmota Himalayana) on the Qinghai-Tibet Plateau of China. Phylogenetic analysis of the 16S rRNA gene and 545 core genes revealed that these two strains belong to the Streptococcus genus. These strains were most closely related to Streptococcus respiraculi HTS25T, Streptococcus cuniculi CCUG 65085T, and Streptococcus marmotae HTS5T. The average nucleotide identity (ANI) and digital DNA‒DNA hybridization (dDDH) were below the threshold for species delineation. The predominant cellular fatty acids (CFAs) in this novel species were C16:0, C18:0, and C18:1ω9c, whereas the primary polar lipids were phosphatidylglycerol (PG), phosphatidylethanolamine (PE) and an unknown phosphoglycolipid (PGL). The optimal growth conditions for the strains were 37 °C, pH 7.0, and 0.5% (w/v) NaCl on brain-heart infusion (BHI) agar supplemented with 5% defibrinated sheep blood. Comparative genomics analyses revealed the potential pathogenicity of strain zg-86T through comparisons with suis subclade strains in terms of virulence factors, pathogen-host interactions (PHIs) and mobile genetic factors (MGEs). Based on the phenotypic characteristics and phylogenetic analyses, we propose that these two isolates represent novel species in the genus Streptococcus, for which the names Streptococcus zhangguiae sp. nov. (the type strain zg-86T=GDMCC 1.1758T=JCM 34273T) is proposed.
Many of the world's freshwater ecosystems suffer from cyanobacteria-mediated blooms and their toxins. However, a mechanistic understanding of why and how Microcystis aeruginosa dominates over other freshwater cyanobacteria during warmer summers is lacking. This paper utilizes comparative genomics with other cyanobacteria and literature reviews to predict the gene functions and genomic architectures of M. aeruginosa based on complete genomes. The primary aim is to understand this species' survival and competitive strategies in warmer freshwater environments. M. aeruginosa strains exhibiting a high proportion of insertion sequences (~ 11%) possess genomic structures with low synteny across different strains. This indicates the occurrence of extensive genomic rearrangements and the presence of many possible diverse genotypes that result in greater population heterogeneities than those in other cyanobacteria in order to increase survivability during rapidly changing and threatening environmental challenges.
Catalase-less M. aeruginosa strains are even vulnerable to low light intensity in freshwater environments with strong ultraviolet radiation. However, they can continuously grow with the help of various defense genes (e.g., egtBD, cruA, and mysABCD) and associated bacteria. The strong defense strategies against biological threats (e.g., antagonistic bacteria, protozoa, and cyanophages) are attributed to dense exopolysaccharide (EPS)-mediated aggregate formation with efficient buoyancy and the secondary metabolites of M. aeruginosa cells. Our review with extensive genome analysis suggests that the ecological vulnerability of M. aeruginosa cells can be overcome by diverse genotypes, secondary defense metabolites, reinforced EPS, and associated bacteria.
We isolated and analyzed a novel, Gram-stain-positive, aerobic, rod-shaped, non-motile actinobacterium, designated as strain ZFBP1038(T), from rock sampled on the north slope of Mount Everest. The growth requirements of this strain were 10-37 °C, pH 4-10, and 0-6% (w/v) NaCl. The sole respiratory quinone was MK-9, and the major fatty acids were anteiso-C(15:0) and iso-C(17:0). Peptidoglycan containing meso-diaminopimelic acid, ribose, and glucose were the major cell wall sugars, while polar lipids included diphosphatidyl glycerol, phosphatidyl glycerol, an unidentified phospholipid, and an unidentified glycolipid. A phylogenetic analysis based on 16S rRNA gene sequences showed that strain ZFBP1038(T) has the highest similarity with Spelaeicoccus albus DSM 26341( T) (96.02%). ZFBP1038(T) formed a distinct monophyletic clade within the family Brevibacteriaceae and was distantly related to the genus Spelaeicoccus. The G + C content of strain ZFBP1038(T) was 63.65 mol% and the genome size was 4.05 Mb.
Digital DNA-DNA hybridization, average nucleotide identity, and average amino acid identity values between the genomes of strain ZFBP1038(T) and representative reference strains were 19.3-25.2, 68.0-71.0, and 52.8-60.1%, respectively.
Phylogenetic, phenotypic, and chemotaxonomic characteristics as well as comparative genome analyses suggested that strain ZFBP1038(T) represents a novel species of a new genus, for which the name Saxibacter gen. nov., sp. nov. was assigned with the type strain Saxibacter everestensis ZFBP1038(T) (= EE 014( T) = GDMCC 1.3024( T) = JCM 35335( T)).
Elevation gradients, often regarded as “natural experiments or laboratories”, can be used to study changes in the distribution
of microbial diversity related to changes in environmental conditions that typically occur over small geographical scales. We
obtained bacterial sequences using MiSeq sequencing and clustered them into operational taxonomic units (OTUs). The total
number of reads obtained by the bacterial 16S rRNA sequencing analysis was 1,090,555, with an average of approximately
45,439 reads per sample collected from various elevations. The current study observed inconsistent bacterial diversity patterns
in samples from the lowest to highest elevations. 983 OTUs were found common among all the elevations. The most
unique OTUs were found in the soil sample from elevation_2, followed by elevation_1. Soil sample collected at elevation_6
had the least unique OTUs. Actinobacteria, Protobacteria, Chloroflexi were found most abundant bacterial phyla in current
study. Ammonium nitrogen (
NH4
+-N), and total phosphate (TP) are the main factors influencing bacterial diversity at elevations_
1. pH was the main factor influencing the bacterial diversity at elevations_2, elevation_3 and elevation_4. Our results
provide new visions on forming and maintaining soil microbial diversity along an elevational gradient and have implications
for microbial responses to environmental change in semiarid mountain ecosystems.
Potyvirids, members of the family Potyviridae, produce the P3N-PIPO protein, which is crucial for the cell-to-cell transport
of viral genomic RNAs. The production of P3N-PIPO requires an adenine (A) insertion caused by RNA polymerase slippage
at a conserved GAA AAA A (
GA6) sequence preceding the PIPO open reading frame. Presently, the slippage rate of
RNA polymerase has been estimated in only a few potyvirids, ranging from 0.8 to 2.1%. In this study, we analyzed publicly
available plant RNA-seq data and identified 19 genome contigs from 13 distinct potyvirids. We further investigated the RNA
polymerase slippage rates at the GA6
motif. Our analysis revealed that the frequency of the A insertion variant ranges from
0.53 to 4.07% in 11 potyviruses (genus Potyvirus). For the two macluraviruses (genus Macluravirus), the frequency of the
A insertion variant was found to be 0.72% and 10.96% respectively. Notably, the estimated RNA polymerase slippage rates
for 12 out of the 13 investigated potyvirids were reported for the first time in this study. Our findings underscore the value of
plant RNA-seq data for quantitative analysis of potyvirid genome variants, specifically at the GA6
slippage site, and contribute
to a more comprehensive understanding of the RNA polymerase slippage phenomenon in potyvirids.
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Microbiome research has been on the rise recently for a more
in-depth understanding of gout. Meanwhile, there is a need to
understand the gut microbiome related to uric acid-lowering
drug resistance. In this study, 16S rRNA gene-based microbiota
analysis was performed for a total of 65 stool samples
from 17 healthy controls and 48 febuxostat-treated gout patients
(including 28 controlled subjects with decreased uric
acid levels and 20 uncontrolled subjects with non-reduced
uric acid levels). Alpha diversity of bacterial community decreased
in the healthy control, controlled, and uncontrolled
groups. In the case of beta diversity, the bacterial community
was significantly different among groups (healthy control, controlled,
and uncontrolled groups). Taxonomic biomarker analysis
revealed the increased population of g-Bifidobacterium
in healthy controls and g-Prevotella in uncontrolled patients.
PCR further confirmed this result at the species level. Additionally,
functional metagenomics predictions led to the exploration
of various functional biomarkers, including purine
metabolism. The results of this study can serve as a basis
for developing potential new strategies for diagnosing and
treating gout from microbiome prospects.
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Three aerobic, Gram-negative, and rod-shaped bacterial strains,
designated strains G4M1T, SM13T, and L12M9T, were isolated
from the gut of Batillaria multiformis, Cellana toreuma, and
Patinopecten yessoensis collected from the Yellow Sea in South
Korea. All the strains grew optimally at 25°C, in the presence
of 2% (w/v) NaCl, and at pH 7. These three strains, which
belonged to the genus Polaribacter in the family Flavobacteriaceae,
shared < 98.8% in 16S rRNA gene sequence and < 86.68%
in whole-genome sequence with each other. Compared with
the type strains of Polaribacter, isolates showed the highest
sequence similarity to P. haliotis KCTC 52418T (< 98.68%),
followed by P. litorisediminis KCTC 52500T (< 98.13%). All
the strains contained MK-6 as their predominant menaquinone
and iso-C15:0 as their major fatty acid. Moreover, all the
strains had phosphatidylethanolamine as their polar lipid
component. In addition, strain G4M1T had two unidentified
lipids and three unidentified aminolipids, strain SM13T had
three unidentified lipids and three unidentified aminolipids,
and strain L12M9T had three unidentified lipids and one unidentified
aminolipid. The DNA G + C contents of strains
G4M1T, SM13T, and L12M9T were 31.0, 30.4, and 29.7 mol%,
respectively. Based on phenotypic, phylogenetic, chemotaxonomic,
and genotypic findings, strains G4M1T (= KCTC 82388T
= DSM 112372T), SM13T (= KCTC 82389T = DSM 112373T),
and L12M9T (= KCTC 62751T = DSM 112374T) were classified
into the genus Polaribacter as the type strains of novel
species, for which the names Polaribacter batillariae sp. nov.,
Polaribacter cellanae sp. nov., and Polaribacter pectinis sp.
nov., respectively, have been proposed.
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To date, all species in the genus Salicibibacter have been isolated
in Korean commercial kimchi. We aimed to describe
the taxonomic characteristics of two strains, NKC5-3T and
NKC21-4T, isolated from commercial kimchi collected from
various regions in the Republic of Korea. Cells of these strains
were rod-shaped, Gram-positive, aerobic, oxidase- and catalase-
positive, non-motile, halophilic, and alkalitolerant. Both
strains, unlike other species of the genus Salicibibacter, could
not grow without NaCl. Strains NKC5-3T and NKC21-4T
could tolerate up to 25.0% (w/v) NaCl (optimum 10%) and
grow at pH 7.0–10.0 (optimum 8.5) and 8.0–9.0 (optimum
8.5), respectively; they showed 97.1% 16S rRNA gene sequence
similarity to each other and were most closely related
to S. kimchii NKC1-1T (97.0% and 96.8% similarity, respectively).
The genome of strain NKC5-3T was nearly 4.6 Mb in
size, with 4,456 protein-coding sequences (CDSs), whereas
NKC21-4T genome was nearly 3.9 Mb in size, with 3,717 CDSs.
OrthoANI values between the novel strains and S. kimchii
NKC1-1T were far lower than the species demarcation threshold.
NKC5-3T and NKC21-4T clustered together to form
branches that were distinct from the other Salicibibacter species.
The major fatty acids in these strains were anteiso-C15:0
and anteiso-C17:0, and the predominant menaquinone was
menaquinone-7. The polar lipids of NKC5-3T included diphosphatidylglycerol
(DPG), phosphatidylglycerol (PG), and
five unidentified phospholipids (PL), and those of NKC21-4T
included DPG, PG, seven unidentified PLs, and an unidentified
lipid. Both isolates had DPG, which is the first case in
the genus Salicibibacter. The genomic G + C content of strains
NKC5-3T and NKC21-4T was 44.7 and 44.9 mol%, respectively.
Based on phenotypic, genomic, phylogenetic, and chemotaxonomic
analyses, strains NKC5-3T (= KACC 22040T
= DSM 111417T) and NKC21-4T (= KACC 22041T = DSM
111418T) represent two novel species of the genus Salicibibacter,
for which the names Salicibibacter cibarius sp. nov.
and Salicibibacter cibi sp. nov. are proposed.
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Newcastle disease, designated a class A disease of poultry by
the Office international des epizooties (OIE), is an acute infection
caused by Newcastle disease virus (NDV). The merging
of the envelope of NDV with the membrane of a target
host cell is the key step in the infection pathway, which is driven
by the concerted action of two glycoproteins: haemagglutinin-
neuraminidase (HN) and fusion (F) protein. When
the HN protein binds to the host cell surface receptor, the F
protein is activated to mediate fusion. The three-dimensional
structure of the F protein has been reported to have low
electron density between the DIII domain and the HRB domain,
and this electron-poor region is defined as the HRB
linker. To clarify the contributing role of the HRB linker in
the NDV F protein-mediated fusion process, 6 single amino
acid mutants were obtained by site-directed mutagenesis of
the HRB linker. The expression of the mutants and their abilities
to mediate fusion were analysed, and the key amino acids
in the HRB linker were identified as L436, E439, I450, and
S453, as they can modulate the fusion ability or expression
of the active form to a certain extent. The data shed light on
the crucial role of the F protein HRB linker in the acquisition
of a normal fusogenic phenotype.
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Intestinal diseases caused by physiological stress have become
a severe public health threat worldwide. Disturbances in the
gut microbiota-host relationship have been associated with
irritable bowel disease (IBD), while melatonin (MT) has antiinflammatory
and antioxidant effects. The objective of this
study was to investigate the mechanisms by which MT-mediated
protection mitigated stress-induced intestinal microbiota
dysbiosis and inflammation. We successfully established a
murine restraint stress model with and without MT supplementation.
Mice subjected to restraint stress had significantly
elevated corticosterone (CORT) levels, decreased MT levels
in their plasma, elevated colonic ROS levels and increased bacterial
abundance, including Bacteroides and Tyzzerella, in
their colon tract, which led to elevated expression of Toll-like
receptor (TLR) 2/4, p-P65 and p-IκB. In contrast, supplementation
with 20 mg/kg MT reversed the elevation of the plasma
CORT levels, downregulated the colon ROS levels and inhibited
the changes in the intestinal microbiota induced by
restraint stress. These effects, in turn, inhibited the activities
of TLR2 and TLR4, p-P65 and p-IκB, and decreased the inflammatory
reaction induced by restraint stress. Our results
suggested that MT may mitigate “restraint stress”-induced
colonic microbiota dysbiosis and intestinal inflammation by
inhibiting the activation of the NF-κB pathway.
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Viruses are found in almost all biomes on Earth, with bacteriophages
(phages) accounting for the majority of viral particles
in most ecosystems. Phages have been isolated from
natural environments using the plaque assay and liquid medium-
based dilution culturing. However, phage cultivation is
restricted by the current limitations in the number of culturable
bacterial strains. Unlike prokaryotes, which possess
universally conserved 16S rRNA genes, phages lack universal
marker genes for viral taxonomy, thus restricting cultureindependent
analyses of viral diversity. To circumvent these
limitations, shotgun viral metagenome sequencing (i.e., metaviromics)
has been developed to enable the extensive sequencing
of a variety of viral particles present in the environment
and is now widely used. Using metaviromics, numerous
studies on viral communities have been conducted in oceans,
lakes, rivers, and soils, resulting in many novel phage sequences.
Furthermore, auxiliary metabolic genes such as ammonic
monooxygenase C and β-lactamase have been discovered
in viral contigs assembled from viral metagenomes.
Current attempts to identify putative bacterial hosts of viral
metagenome sequences based on sequence homology have
been limited due to viral sequence variations. Therefore, culture-
independent approaches have been developed to predict
bacterial hosts using single-cell genomics and fluorescentlabeling.
This review focuses on recent viral metagenome
studies conducted in natural environments, especially in aquatic
ecosystems, and their contributions to phage ecology.
Here, we concluded that although metaviromics is a key tool
for the study of viral ecology, this approach must be supplemented
with phage-host identification, which in turn requires
the cultivation of phage-bacteria systems.
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Long-read powered viral metagenomics in the oligotrophic Sargasso Sea Joanna Warwick-Dugdale, Funing Tian, Michelle L. Michelsen, Dylan R. Cronin, Karen Moore, Audrey Farbos, Lauren Chittick, Ashley Bell, Ahmed A. Zayed, Holger H. Buchholz, Luis M. Bolanos, Rachel J. Parsons, Michael J. Allen, Matthew B. Sullivan, Ben Tempe Nature Communications.2024;[Epub] CrossRef
Tools and methodology to in silico phage discovery in freshwater environments Carlos Willian Dias Dantas, David Tavares Martins, Wylerson Guimarães Nogueira, Oscar Victor Cardenas Alegria, Rommel Thiago Jucá Ramos Frontiers in Microbiology.2024;[Epub] CrossRef
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Metaviromics analysis of marine biofilm reveals a glycoside hydrolase endolysin with high specificity towards Acinetobacter baumannii Georgios E. Premetis, Nikolaos D. Georgakis, Angeliki Stathi, Nikolaos E. Labrou Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics.2023; 1871(4): 140918. CrossRef
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Ammonia oxidation, performed by ammonia-oxidizing archaea
(AOA) and bacteria (AOB), plays a critical role in the cycle
of nitrogen in the ocean. For now, environmental variables
controlling distribution of ammonia-oxidizing microbes are
still largely unknown in oceanic environments. In this study,
we used real-time quantitative PCR and high-throughput sequencing methods to investigate the abundance and diversity
of AOA and AOB from sediment and water in Zhanjiang Bay.
Phylogenic analysis revealed that the majority of AOA amoA
sequences in water and sediment were affiliated with the genus
Nitrosopumilus, whereas the Nitrosotalea cluster was only detected
with low abundance in water. Nitrosomonas and Nitrosospira
dominated AOB amoA sequences in water and sediment,
respectively. The amoA copy numbers of both AOA and
AOB varied significantly with month for both sediment and
water. When water and sediment temperature dropped to 17–
20°C in December and February, respectively, the copy number
of AOB amoA genes increased markedly and was much
higher than for AOA amoA genes. Also, AOA abundance in
water peaked in December when water temperature was lowest
(17–20°C). Stepwise multiple regression analyses revealed that
temperature was the most key factor driving monthly changes
of AOA or AOB abundance. It is inferred that low water temperature
may inhibit growth of phytoplankton and other microbes
and so reduce competition for a common substrate,
ammonium.
6S RNA, a conserved and abundant small non-coding RNA
found in most bacteria, regulates gene expression by inhibiting
RNA polymerase (RNAP) holoenzyme. 6S RNAs from
α-proteobacteria have been studied poorly so far. Here, we
present a first in-depth analysis of 6S RNAs from two α-proteobacteria
species, Bradyrhizobium japonicum and Sinorhizobium
meliloti. Although both belong to the order Rhizobiales
and are typical nitrogen-fixing symbionts of legumes,
their 6S RNA expression profiles were found to differ: B. japonicum
6S RNA accumulated in the stationary phase, thus
being reminiscent of Escherichia coli 6S RNA, whereas S. meliloti
6S RNA level peaked at the transition to the stationary
phase, similarly to Rhodobacter sphaeroides 6S RNA. We demonstrated
in vitro that both RNAs have hallmarks of 6S
RNAs: they bind to the σ70-type RNAP holoenzyme and serve
as templates for de novo transcription of so-called product
RNAs (pRNAs) ranging in length from ~13 to 24 nucleotides,
with further evidence of the synthesis of even longer pRNAs.
Likewise, stably bound pRNAs were found to rearrange the
6S RNA structure to induce its dissociation from RNAP.
Compared with B. japonicum 6S RNA, considerable conformational
heterogeneity was observed for S. meliloti 6S RNA
and its complexes with pRNAs, even though the two 6S RNAs
share ~75% sequence identity. Overall, our findings suggest
that the two rhizobial 6S RNAs have diverged with respect to
their regulatory impact on gene expression throughout the
bacterial life cycle.
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Detection of Small Products of Transcription from 6S RNA (pRNA) by “Mirror-Like” Northern Blot Hybridization O. Y. Burenina, T. S. Oretskaya, E. A. Kubareva Russian Journal of Bioorganic Chemistry.2021; 47(2): 478. CrossRef
Escherichia coli (E. coli) infection is very common among
young growing animals, and zinc supplementation is often
used to alleviate inflammation induced by this disease. Therefore,
the objective of this study was to evaluate whether chitosan-
chelated zinc (CS-Zn) supplementation could attenuate
gut injury induced by E. coli challenge and to explore how CSZn
modulates cecal microbiota and alleviates intestinal inflammation
in weaned rats challenged with E. coli. 36 weaned
rats (55.65 ± 2.18 g of BW, n = 12) were divided into three
treatment groups consisting of unchallenged rats fed a basal
diet (Control) and two groups of rats challenged with E. coli
and fed a basal diet or a diet containing 640 mg/kg CS-Zn
(E. coli + CS-Zn, containing 50 mg/kg Zn) for a 14-day experiment.
On days 10 to 12, each rat was given 4 ml of E. coli
solution with a total bacteria count of 1010 CFU by oral gavage
daily or normal saline of equal dosage. CS-Zn supplementation
mitigated intestinal morphology impairment (e.g.
higher crypt depth and lower macroscopic damage index)
induced by E. coli challenge (P < 0.05), and alleviated the increase
of Myeloperoxidase (MPO) activity after E. coli challenge
(P < 0.05). 16S rRNA sequencing analyses revealed that
E. coli challenge significantly increased the abundance of Verrucomicrobia
and E. coli (P < 0.05). However, CS-Zn supplementation
increased the abundance of Lactobacillus and decreased
the relative abundance of Proteobacteria, Desulfovibrio
and E. coli (P < 0.05). The concentrations of butyrate in
the cecal digesta, which decreased due to the challenge, were
higher in the E. coli + CS-Zn group (P < 0.05). In addition,
CS-Zn supplementation significantly prevented the elevation
of pro-inflammatory cytokines IL-6 concentration and upregulated
the level of anti-inflammatory cytokines IL-10 in
cecal mucosa induced by E. coli infection (P < 0.05). In conclusion,
these results indicate that CS-Zn produces beneficial
effects in alleviating gut mucosal injury of E. coli challenged
rats by enhancing the intestinal morphology and modulating
cecal bacterial composition, as well as attenuating inflammatory
response.
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