16S rDNA Research Articles

Microbiological Quality Assessment of Some Commercially Available Breads.

Bread is a staple, energy-rich food for people of all ages, so quality is important to consumers. In our region, most of the commercially available bread, whether packaged or unpackaged, is produced by local bakeries, so monitoring microbial levels and the types of microbes present on bread can help to draw attention to protect the final product. It can also help to ensure the food safety, quality, and shelf life of bread. The freshly baked product is microbiologically sterile. Post-process contamination affects the microbial load of bread. In this study, the microbial load of 30 different commercial bread crumbs and crusts was determined. The different types of bread with different compositions were analyzed for total viable bacteria, Escherichia coli, Staphylococcus aureus, aerobic and anaerobic spore-forming bacteria, and culturable microscopic fungi. The K-means clustering algorithm was used to cluster the different types of bread based on the number of aerobic mesophilic bacteria. Significant differences (p < 0.05) were found in the total viable bacterial count for bread crusts and crumbs. The bacterial count of bread varied between 10.00 ± 0.00-395.00 ± 52.4 CFU/g for bread crusts and 10.00 ± 0.0-310.67 ± 94 CFU/g for bread crumbs. The results of 16S rDNA sequence analysis showed that the most frequently occurring bacterial species belonged to the genus Bacillus, but species of the genus Staphylococcus were also present. Chryseobacterium spp. predominated on multigrain bread, Marinilactobacillus spp. on rustic potato bread, and Staphylococcus warneri on sliced brown potato bread. The results contribute to a better understanding of the microbial dynamics in locally produced breads from the Eastern Carpathians of Transylvania, with the aim of improving food safety, quality control, and consumer protection.

Integration of host gene regulation and oral microbiome reveals the influences of smoking during the development of oral squamous cell carcinoma.

This study aims to investigate the regulation of host gene transcription and microbial changes during the development of oral squamous cell carcinoma (OSCC) associated with smoking. The OSCC mouse model and smoking mouse model were established using 200 μg/mL 4-nitroquinoline-1-oxide (4NQO) in drinking water and exposure to cigarette smoke (four cigarettes per session, once a day, 5 days a week). Tongue tissues were harvested at 4 weeks and 16 weeks. Histopathological changes were evaluated using hematoxylin and eosin staining and Ki67 staining. RNA sequencing was performed on the mouse tongue tissues to identify differentially expressed genes (DEGs), and the results were validated by RT-PCR and immunohistochemistry. 16S rDNA sequencing was used to analyze changes in the oral microbiota during the early development of OSCC, identifying differentially abundant taxa associated with smoking. Finally, associations between the relative abundances of the oral microbiome and host gene expression were modeled using the Origin software. DEGs associated with smoking during the development of OSCC were identified. There were 12 upregulated genes, including NR4A3 and PPP1R3C, and 23 downregulated genes, including CD74 and ANKRD1. These genes were enriched in functions related to the signal transduction of cellular processes such as inflammation, differentiation, immunity, and PI3K/AKT, NF-κB signaling pathways. 4NQO and smoking treatment decreased oral microbial diversity and reduced the abundance of Bacteroidetes, Proteobacteria, and Lactobacillus but increased the abundance of Staphylococcus. Integrative analysis showed that the expression of CD74 was positively correlated with the relative abundance of Lactobacillus, while PPP1R3C was negatively correlated with Bacteroidota. In addition to characterizing host gene expression and the oral microbiome, our study explored the potential role of host-microbiome interactions in the development of OSCC. These findings enhance our understanding of smoking-related OSCC occurrence and development, providing new insights for its prevention.

Effect of xylo-oligosaccharides on intestinal bacterial diversity in mice with spleen deficiency constipation.

To explore the effect of xylo-oligosaccharides on intestinal bacterial diversity in mice with spleen deficiency constipation. The 16S rDNA sequencing was used to identify microbiota composition in four groups, including the normal group (NG), the model group with spleen-deficiency constipation (SDC), XOS treated groups that include XOS1 groups treated XOS 0.05 g/mL•d, and XOS2 group treated XOS 0.1 g/mL•d. Chao1 and Shannon were used to conduct gut microbes diversity analysis. Linear discriminant analysis coupled with effect size measurements (LEfSe) was used to identify signature gut microbiota, and phylogenetic investigation of communities by reconstruction of unobserved states (PICRUSt) was used to predict the function of altered gut microbiota. Veen map indicated 245 common OTUs were identified from four groups. Especially, 9, 3, 0, and 19 unique OTUs were identified in NG, SDC, XOS1, and XOS2 groups, respectively. The Shannon index was evidently higher in NG group than in the other three groups (p < 0.05). We identified the occurrence of dominant bacterial groups including Bacteroidetes (25.5 ~ 49.9%), Firmicutes (25.4 ~ 39.3%), Proteobacteria (12.5 ~ 24.9%), Deferribacteres (1.6 ~ 19.2%), Cyanobacteria (0.3 ~ 1.8%), Verrucomicrobia (0.02 ~ 1.6%), Actinobacteria (0.01 ~ 0.5%), and Tenericutes (0.03 ~ 0.09%) at the four groups. The XOS2 group was characterized by a higher abundance of Peptostreptococcaceae, Intestinibacter, Aerococcaceae, and Facklamia. XOS1 group enriched in Deferribacteres, Mucispirillum, Deferribacterales, Deferribacteres, Lachnoclostridium, Rhodospirillaceae, and Rhodospirillales. Meanwhile, the SDC mice showed dramatic enrichment in Rikenellaceae, Lachnospiraceae, Rikenellaceae, Roseburia, and Alistipes, which were highly abundant in the NG group. XOS fed-mice evidently increase Deferribcteres abundance compared with NG and SDC groups. However, the abundance of Rikenellaceae was significantly reduced in XOS1 and XOS2 groups compared with NG and SDC groups. We identified that altered gut microbiotas by XOS treatment were associated with various metabolic pathways, including organismal systems, metabolism, human diseases, genetic information processing, and cellular processes. Our research indicated that XOS has the potential to recover intestinal bacteria and contribute to the treatment of spleen deficiency constipation.

The histone chaperones ASF1 and HIRA are required for telomere length and 45S rDNA copy number homeostasis.

Genome stability is significantly influenced by the precise coordination of chromatin complexes that facilitate the loading and eviction of histones from chromatin during replication, transcription, and DNA repair processes. In this study, we investigate the role of the Arabidopsis H3 histone chaperones ANTI-SILENCING FUNCTION 1 (ASF1) and HISTONE REGULATOR A (HIRA) in the maintenance of telomeres and 45S rDNA loci, genomic sites that are particularly susceptible to changes in the chromatin structure. We find that both ASF1 and HIRA are essential for telomere length regulation, as telomeres are significantly shorter in asf1a1b and hira mutants. However, these shorter telomeres remain localized around the nucleolus and exhibit a comparable relative H3 occupancy to the wild type. In addition to regulating telomere length, ASF1 and HIRA contribute to silencing 45S rRNA genes and affect their copy number. Besides, ASF1 supports global heterochromatin maintenance. Our findings also indicate that ASF1 transiently binds to the TELOMERE REPEAT BINDING 1 protein and the N terminus of telomerase in vivo, suggesting a physical link between the ASF1 histone chaperone and the telomere maintenance machinery.

Intestinal endogenous metabolites affect neuroinflammation in 5×FAD mice by mediating “gut-brain” axis and the intervention with Chinese Medicine

BackgroundEmerging evidence suggested the association between gut dysbiosis and Alzheimer’s disease (AD) progression. However, it remained unclear how the gut microbiome and neuroinflammation in the brain mutually interact or how these interactions affect brain functioning and cognition. Here we hypothesized that “gut-brain” axis mediated by microbial derived metabolites was expected to novel breakthroughs in the fields of AD research and development.MethodsMultiple technologies, such as immunofluorescence, 16s rDNA sequencing, mass spectrometry-based metabolomics (LC-QQQ-MS and GC-MS), were used to reveal potential link between gut microbiota and the metabolism and cognition of the host.ResultsMicrobial depletion induced by the antibiotics mix (ABX) verified that “gut-brain” can transmit information bidirectionally. Short-chain fatty acid-producing (SCFAs-producing) bacteria and amino acid-producing bacteria fluctuated greatly in 5×FAD mice, especially the reduction sharply of the Bifidobacteriaceae and the increase of the Lachnospiraceae family. Concentrations of several Tryptophan-kynurenine intermediates, lactic acid, CD4+ cell, and CD8+ cells were higher in serum of 5×FAD mice, whilst TCA cycle intermediates and Th1/Th2 were lower. In addition, the levels of iso-butyric acid (IBA) in feces, serum, and brain of 5×FAD mice were increased compared with WT-M mice, especially in serum. And IBA in the brain was positively correlated with Aβ and proinflammatory factors.ConclusionTogether, our finding highlighted that the alternation in gut microbiota affected the effective communication between the “gut-brain” axis in 5×FAD mice by regulating the immune system, carbohydrate, and energy metabolism.

Screening and application of functional autochthonous starter culture from cured meat, which can reduce nitrite content.

Cured meat is a fermented meat product from the traditional Chinese culture made by natural fermentation. In this study, five bacteria strains were screened from cured meat using 16S rDNA technology, and a functional local starter was selected, which was applied to the production of cured meat to standardize the production of cured meat and improve the quality of cured meat. By studying the fermentation characteristics of strain these strains, this study found that the fermentation characteristics of L. mesenteroides and S. lactis are ideal. L. mesenteroides and S. lactis were used as starter cultures in fermented bacon. Then, this study compared the quality of fermented beef with Sichuan bacon, Hunan bacon, and Xinyang bacon. The results suggested that L. mesenteroides and S. lactis can improve the sensory and texture properties of the products and reduce the moisture content, water activity, pH value, and protein content of fermented beef products. More importantly, L. mesenteroides can significantly reduce the nitrite content (25.34%) and nitrosamine content (29.69%) in fermented beef, which provides an excellent guarantee for the safety of cured meat. In this study, a functional fermentation strain-L. mesenteroides could degrade the nitrite content of fermented meat products and improve their sensory and textural properties-was screened to provide some reference value for the later development of functional strains suitable for fermented meat products.

Potential of novel Rhizobium strains as biofertilizer for improved groundnut (Arachis hypogaea L.) cultivation in Eastern India

ABSTRACT Comprehensive assessments were conducted on eight fast growing, novel Rhizobium isolates, short listed from a preliminary selection of 68 isolates obtained from root nodules of groundnut plants grown in diverse agro-climatic regions of eastern India. Additionally, two conventional rhizobia strains, viz. IGR6 and GN2, (collected from the Nodule Research Center, BCKV), were utilized for comparative analysis. Cluster analysis, based on carbon utilization and biochemical parameters, grouped all the test isolates with an average similarity level of 73%. The ability of the isolates to utilize a wide range of carbon sources, including hexoses, pentoses, and disaccharides, confirms their fast-growing nature. Further analysis through 16S rDNA extraction and nucleotide sequencing revealed that the promising fresh isolates exhibited 95–99% similarity to known Rhizobium species in public databases. When evaluated in pot culture experiments using new alluvial soil from the Gangetic Plains in eastern India, the native fresh isolate NRA1(PP355674) outperformed other isolates and conventional strains in terms of nodulation efficiency, plant dry matter production, and plant nitrogen uptake. This study highlights the significant potential of native Rhizobium isolates to surpass conventionally used strains in biological nitrogen fixation, supporting the development of sustainable, environmentally-friendly, and cost-effective agricultural practices.

Anaplasma platys and Rickettsia massiliae in Rhipicephalus sanguineus sensu stricto ticks collected on dogs in the Patagonian region of Argentina.

The aim of this study was to examine the presence of tick-borne rickettsial bacteria in Rhipicephalus sanguineus sensu stricto ticks collected from dogs in the Patagonian region of Argentina. Fourteen stray dogs from Valcheta, Río Negro province, Argentina were examined for the presence of R. sanguineus s.s. ticks. Ninety ticks were collected and identified to species level. DNA was extracted and analysed by conventional PCR assays for the presence of tick-borne bacteria belonging to the genera Anaplasma, Ehrlichia and Rickettsia. Thirty-three tick pools were tested by different PCR assays of which 3 were positive for Anaplasmataceae bacteria. From the 3 Anaplasmataceae positive samples, 2 partial 16S rDNA sequences were generated and belonging to Anaplasma platys, the causative agent of canine cyclic thrombocytopenia. Two tick samples were positive in Rickettsia specific PCR assays and were identified by phylogenetic analysis as Rickettsia massiliae, a member of the spotted fever group rickettsiae. The results of this study demonstrate the molecular detection of 2 rickettsial bacteria in R. sanguineus s.s. in a region of Argentina where no data were available so far.

Isolation and identification of cellulose decomposition bacteria from the soil of Yunnan province, China

In this experiment sodium carboxymethyl cellulose (CMC-Na) was used as the only carbon source to isolate cellulose decomposing bacteria from the soil of Qiongzhusi Forest Park in Kunming, Yunnan Province. The ratio of hydrolytic circle diameter and colony diameter formed by the strain on cellulose Congo red medium was determined for primary screening, and then the filter paper disintegration test and CMC enzyme activity assay were used for rescreen. A cellulose decomposing bacterium (HS-2) with high enzyme activity was obtained, and its enzyme activity was up to 16.42 U/ml. (U means the unit of activity of enzymes, activity, unit) Gram stain and Spore stain reaction showed that HS-2 strain was Gram-positive Bacillus. The Bacillus strain was identified as Bacillus pumilus on the basis of physiological and biochemical tests, and 16S rDNA sequence analysis. Bangladesh J. Bot. 52(3): 635-641, 2024 (September) Special

Ameliorated Viability of Lactic Acid Bacteria in Fruit Juice Isolated from Indigenous Dahi with Prebiotics (Asparagus falcatus and Zingiber officinale)

Dahi is a fermented milk product containing probiotic lactic acid bacteria. This study aimed to isolate, identify, and characterize lactic acid-producing bacteria from native Dahi and evaluate their viability in orange juice using natural prebiotics. Dahi samples were obtained from local shops in Chattogram and Bogura, Bangladesh. Lactic acid-producing bacteria were isolated using MRS (de Mann Rogosa and Sharpe) medium. The isolated bacteria were identified through colony morphology, biochemical tests, and probiotic characteristics. Molecular identification was performed using polymerase chain reaction (PCR) targeting conserved 16S rDNA regions. Isolates of the genus Lactobacillus and Lactococcus lactis sp. Lactis were confirmed and used to develop probiotic orange juice. Prebiotics (Asparagus falcatus and Zingiber officinale) were added to the juice to support probiotic growth. The inoculated cell’s viability and the juice’s physicochemical parameters were evaluated during fermentation (48 hours) and storage (28 days). All fruit juice samples showed a mean number of viable cells of at least 1×105 CFU/mL during the 48-hour fermentation and 28-day storage in the refrigerator. Using natural prebiotics positively affected the survival of lactic acid bacteria, as demonstrated by bacterial colony growth on Petri dishes. Developing probiotic fruit juice enriched with prebiotics could be an effective alternative for individuals allergic or intolerant to milk-based products. Incorporating lactic acid bacteria from native Dahi into orange juice, combined with natural prebiotics, resulted in viable probiotic cells throughout fermentation and storage.

Entamoeba muris mitigates metabolic consequences of high-fat diet in mice

ABSTRACT Metabolic syndrome (MetS) is a cluster of several human conditions including abdominal obesity, hypertension, dyslipidemia, and hyperglycemia, all of which are risk factors of type 2 diabetes, cardiovascular disease, and metabolic dysfunction-associated steatotic liver disease (MASLD). Dietary pattern is a well-recognized MetS risk factor, but additional changes related to the modern Western life-style may also contribute to MetS. Here we hypothesize that the disappearance of amoebas in the gut plays a role in the emergence of MetS in association with dietary changes. Four groups of C57B/6J mice fed with a high-fat diet (HFD) or a normal diet (ND) were colonized or not with Entamoeba muris, a commensal amoeba. Seventy days after inoculation, cecal microbiota, and bile acid compositions were analyzed by high-throughput sequencing of 16S rDNA and mass spectrometry, respectively. Cytokine concentrations were measured in the gut, liver, and mesenteric fat looking for low-grade inflammation. The impact of HFD on liver metabolic dysfunction was explored by Oil Red O staining, triglycerides, cholesterol concentrations, and the expression of genes involved in β-oxidation and lipogenesis. Colonization with E. muris had a beneficial impact, with a reduction in dysbiosis, lower levels of fecal secondary bile acids, and an improvement in hepatic steatosis, arguing for a protective role of commensal amoebas in MetS and more specifically HFD-associated MASLD.

Fluorescence in situ hybridization analysis of Oligonucleotide 5S rDNA, 45S rDNA, and (TTTAGGG)3 locations in Gloriosa superba L.

Gloriosa superba L. is a horticulturally and medicinally important plant native to Africa. However, the few cytogenetic studies of the species are mainly focused on chromosome counting and chromosome morphology-based karyotyping. Fluorescence in situ hybridization (FISH) is a powerful tool for the detection of DNA repetitive elements in a specific region of a chromosome. Here, detailed karyotypes of G. superba were constructed by FISH using 5S and 45S rDNAs, and telomeric repeat (TTTAGGG)3 oligonucleotides. Twenty-two chromosomes were observed. Two 5S rDNA hybridization signals were detected in the proximal regions of the short arms of one pair of chromosomes, which were adjacent to the (TTTAGGG)3 terminal signals. Four 45S rDNA signals were detected near the centromere region of the short arm of the four chromosomes, but one of these was very weak and almost undetectable compared to the others. Telomeric repeat hybridization signals were distributed at the terminal region of each chromosome. The chromosomes displayed were intact and the chromosome counts were accurate. Chromosome length ranged from 3.46 to 9.31 μm. These results will facilitate the cytogenetic mapping of other major repeats, thus contributing to an improved understanding of the G. superba genome structure and evolutionary history.

Two new species of genus Labronema Thorne, 1939 (Nematoda, Dorylaimidae) from natural parks of Vietnam with an identification key to the species with a medium-sized odontostyle.

Labronema Thorne, 1939 is a large and diverse dorylaimid genus with complicated taxonomy. Two new species, Labronemaporosum sp. nov. and Labronemabidoupense sp. nov. from natural habitats in Vietnam were characterised both morphologically and molecularly (18S rDNA and 28 rDNA), and line drawings and microphotographs are provided. Phylogenetic analyses showed that the new species clustered together with Labronemaferox Thorne, 1939, the type species of the genus. The two new taxa belong to a group of Labronema species with medium body (L = 1.5-2.5 mm) and odontostyle (31-39 μm) length, and a lip region offset by a constriction. Based on morphological and molecular evidence, this study shows that the populations from Vietnam previously identified as L.glandosum Rahman, Jairajpuri, Ahmad & Ahmad, 1986 in fact represent L.porosum sp. nov. Evolutionary relationships of Labronema species are discussed. A key to the species of Labronema with a medium-sized odontostyle (31-39 μm) is provided as well as a list of the species of the genus considered valid.

Three new species of the genus ‘Carteria’ (Chlamydomonadales, Chlorophyceae) with the proposal of correct generic names

ABSTRACT ‘Carteria’ is a genus of quadriflagellate chlamydomonadalean green algae, however this genus is not thought to be a monophyletic group. In this study, we investigated nine strains of ‘Carteria’ newly isolated from China by morphological comparisons and molecular phylogeny of 18S rDNA and rbcL genes. Five strains could be identified as existing ‘Carteria’ species. Four strains with clearly different morphology and independent phylogenetic positions compared with the described ‘Carteria’ species were identified as three new species. Since the original type species, Carteria cordiformis, has been recognized as a prasinophyte alga, the generic name Carteria cannot be applied to other species previously assigned to ‘Carteria’. Six ‘Carteria’ species (including three new species) and one Pseudocarteria species of Carteria-Clade I were transferred to the genus Corbierea, and the description of this genus was emended. A new genus, Staurocarteria gen. nov. was proposed for Carteria-Clade II.

LysoPE mediated by respiratory microorganism Aeromicrobium camelliae alleviates H9N2 challenge in mice

Influenza remains a severe respiratory illness that poses significant global health threats. Recent studies have identified distinct microbial communities within the respiratory tract, from nostrils to alveoli. This research explores specific anti-influenza respiratory microbes using a mouse model supported by 16S rDNA sequencing and untargeted metabolomics. The study found that transferring respiratory microbes from mice that survived H9N2 influenza to antibiotic-treated mice enhanced infection resistance. Notably, the levels of Aeromicrobium were significantly higher in the surviving mice. Mice pre-treated with antibiotics and then inoculated with Aeromicrobium camelliae showed reduced infection severity, as evidenced by decreased weight loss, higher survival rates, and lower lung viral titres. Metabolomic analysis revealed elevated LysoPE (16:0) levels in mildly infected mice. In vivo and in vitro experiments indicated that LysoPE (16:0) suppresses inducible nitric oxide synthase (INOS) and cyclooxygenase-2 (COX2) expression, enhancing anti-influenza defences. Our findings suggest that Aeromicrobium camelliae could serve as a potential agent for influenza prevention and a prognostic marker for influenza outcomes.

New Molecular Phylogenetic Evidence Confirms Independent Origin of Coxal Combs in the Families of the 'Cydnoid' Complex (Hemiptera: Heteroptera: Pentatomoidea).

Coxal combs, found only in members of the 'cydnoid' complex (comprising four families: Cydnidae, Parastrachiidae, Thaumastellidae, and Thyreocoridae) within the superfamily Pentatomoidea, have long been regarded as a character confirming their close evolutionary relationship. However, many studies have demonstrated that these four families are phylogenetically distant. Others have been treated as subfamilies of the broadly defined Cydnidae, with the coxal combs as the only character linking them. This is the first study on the origin of coxal combs in species of all families and subfamilies that represent the broadly conceived Cydnidae (69 species in 39 genera). Moreover, this study presents the first 16S rDNA gene sequences providing a basis for such analyses. The analyses included DNA sequences of 62 species in 34 genera of Cydnidae sensu stricto, three species in two genera of Thyreocoridae, two species in two genera of Parastrachiidae, and two species in one genus of Thaumastellidae. The sequence analysis in the family Cydnidae covered 35 species representing 19 genera of the subfamily Cydninae, 16 species in eight genera of the subfamily Sehirinae, five species in two genera of Amnestinae, three species in three genera of Garsauriinae, two species in one genus of Cephalocteinae, and one species of Amaurocorinae. The results of our study demonstrate the independent origin of coxal combs in taxa of the 'cydnoid' complex within the superfamily Pentatomoidea. They confirm the polyphyly not only of the entire 'cydnoid' complex but also of the family Cydnidae itself.

Integrated serum pharmacochemistry, 16S rDNA sequencing, and metabolomics to reveal the material basis and mechanism of Shouhui Tongbian capsule against diphenoxylate-induced slow transit constipation in rats

BackgroundSlow transit constipation (STC) is highly prevalent and has rising incidence. Shouhui Tongbian capsule (SHTB) is a traditional Chinese Medicine formula with extensive and highly efficacious usage in STC treatment, however, its mechanism of action, especially the regulation of microbiome and lipid metabolites, remains unclear.MethodsAfter quality control of SHTB using LC‒MS to obtain its material basis, we tried to elucidate the cohesive modulatory network of SHTB against STC using hyphenated methods from microbiomics, lipidomics, mass spectrometry imaging (MSI) and molecular methods.ResultsSHTB could repair intestinal barrier damage, reduce systemic inflammation and increase intestinal motility in a diphenoxylate-induced STC rat model. Based on 16S rDNA sequencing results, SHTB rehabilitated the abnormal changes in Alloprevotella, Coprococcus, Marvinbryantia, etc., which were associated with STC symptoms. Meanwhile, microbial functional prediction showed that lipid metabolism was improved with SHTB administration. The differential lipids, including fatty acids, lysophosphatidylcholine, phosphatidylcholine, sphingomyelin triglyceride and ceramide, that are closely related to STC disease and SHTB efficacy. Furthermore, SHTB significantly reversed the abnormal expression of these key target enzymes in colon samples, including CTP-phosphocholine cytidylyltransferase, CTP-phosphoethanolamine cytidylyltransferase, phosphatidic acid phosphatase, acid sphingomyelinase etc.ConclusionsCombined analysis demonstrated that SHTB reducing lipid accumulation and recovery of intestinal microbial homeostasis was the critical mechanism by which SHTB treats STC.

Lactococcus garvieae as a Novel Pathogen in Cultured Pufferfish (Takifugu obscurus) in China

In October 2023, a disease outbreak in pufferfish (Takifugu obscurus) farms in Zhongshan City, Guangdong, China, caused high mortality. Diseased fish (mean length: 15 ± 1 cm) exhibited swimming disorders, fin rot, hemorrhage, and an enlarged spleen. Histopathological observations generally revealed inflammation, necrosis, and congestion in the spleen, kidneys, and brain tissues. The most severe pathological changes included interstitial edema and tubular atrophy in the kidneys, hemosiderin deposition in the spleen, massive red blood cell infiltration, and a decrease in lymphocytes. A single strain of bacteria (Tol-1) was isolated from the diseased pufferfish and identified as a Gram-positive streptococcus strain, exhibiting α-hemolysis on sheep blood agar plates. Through biochemical characterization, 16S rDNA sequencing, morphological analysis, and specific primer-based identification, the Tol-1 strain was identified as Lactococcus garvieae, serotype I. Antimicrobial susceptibility testing indicated that Tol-1 was sensitive to Chloramphenicol, Ampicillin, Cephalexin, and Doxycycline, but resistant to Kanamycin, Gentamicin and Ciprofloxacin. In addition, 15 common virulence factors were detected in the Tol-1 strain, including adhPav, adhPsaA, adhC I–II, adh, and hly 1–3. Pufferfish (mean length: 17 ± 1 cm) subjected to artificial infection via intraperitoneal injection (IP) with the Tol-1 strain exhibited clinical symptoms and histopathological damage similar to those observed in naturally infected fish. An infection dose of 1 × 105 CFU/fish resulted in 80% mortality. The study fulfilled Koch’s postulates, indicating that the disease outbreak in pufferfish was caused by L. garvieae, which exhibited a high mortality rate in pufferfish despite the subtle clinical symptoms. These results serve as a warning for pufferfish farming areas and provide a scientific basis for future prevention and control efforts.

Variations in gut microbiota composition and reproductive hormone levels between laying and broody Muscovy ducks

High broodiness in Muscovy ducks impedes animal husbandry growth. The interaction between endocrine hormones and gut microbiota has been proven to play a crucial role in reproductive performance, and whether it can regulate the broody behavior of Muscovy ducks requires further research. Nine laying ducks (Laying group) and nine broody ducks (Broodiness group) were selected. Corresponding serum, ileum, and cecum chyme were collected for further research. The results showed that, compared to the laying group, the serum concentration of prolactin decreased, while the levels of Mullerian inhibiting substance, follicle-stimulating hormone, and follistatin increased in the broodiness group (P < 0.05). 16S rDNA sequencing showed that, the broodiness group exhibited lower abundance levels of Rothia, Streptococcus, and Lactobacillus, whereas the abundance of Turicibacter, Aliicoccus, and Facklamia was higher in the ileum compared to the laying group (P < 0.05). In the cecum, the broodiness group exhibits a significant reduction in the abundance of Butyricicoccus and unclassified_f_Rikenellaceae, while the abundance of Christensenellaceae_R-7_group, Ruminococcus_torques_group, Parabacteroides, norank_f_Oscillospiraceae, Cloacibacillus, Sellimonas, Shuttleworthia, norank_f_UCG-010, unclassified_f_Lachnospiraceae, Oscillospira, Synergistes, Family_XIII_AD3011_group and Eubacterium_nodatum_group is higher compared to the laying group. A Spearman correlation analysis reveals that both in the ileum and cecum, serum hormones exhibit significant correlations with the top 20 abundant intestinal microbial genera. Among these, serum follistatin has most entries of significant correlations with the detected microbial genera (P < 0.05). In conclusion, the broody behavior of Muscovy ducks can be modulated by the interaction between hormones and gut microbiota. Notably, the relationship between Follistatin and the composition of gut microbiota, specifically Firmicutes, is the most prominent.

Lactobacillus fermentum SNR1: Probiotic efficiency and characterization of the antimicrobial peptide against dental caries and foodborne pathogens

In this study, fermented rice gruel, a traditional food in South Telangana, India, was used as a source for the isolation of biologically significant probiotics. A total of 56 morphologically diverse strains were isolated and screened for their ability to inhibit dental biofilms as well as foodborne and aquaculture bacterial pathogens. Among these, isolate SNR1 was selected for further characterization due to its potential antimicrobial activity against bacterial pathogens such as Streptococcus mutans, Listeria monocytogenes, Staphylococcus aureus, Vibrio parahaemolyticus and Salmonella typhi. The results of 16S rDNA sequencing showed that isolate SNR1 was closely related to Lactobacillus fermentum strains from different sources. The bacteriocin peptide with potential activity was extracted and purified from the cell-free supernatant using ammonium sulphate precipitation, gel permeation chromatography and reversed-phase high performance liquid chromatography (RP-HPLC), and was named peptide SNR1. Tricine SDS-PAGE followed by MALDI-TOF-MS analysis revealed that the purified peptide had a molecular mass of 4.33 kDa, with the N-terminal sequence identified as MPTTSHFHLSPQGK. The purified peptide SNR1 showed good stability with various physiological enzymes, temperatures and pH conditions.