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BMC Veterinary Research Dec 2022Bacterial pathogens are a great threat to fish production. Gram-negative bacteria are among the major bacterial fish pathogens and are zoonotic with the potential to...
Isolation and identification of major bacteria from three Ethiopian rift valley lakes live and processed fish, and water samples: implications in sanitary system of fish products.
Bacterial pathogens are a great threat to fish production. Gram-negative bacteria are among the major bacterial fish pathogens and are zoonotic with the potential to infect humans. This cross-sectional study was conducted to isolate and identify major gram-negative bacteria from live and processed fish, and water samples from Lakes Hawassa, Langanoo and Ziway. A total of 674 different types of samples: 630 tissue samples (210 samples for each intestine, Kkidney and liver collected from 210 live fish (Oreochromis niloticus, Cyprinus carpio and Clarias gariepinus), 20 processed fish samples from lake Ziway fish processing center and 24 lake water samples were included in the study from each lake. The mean values of pH, temperature, dissolved oxygen and nitrate in all water samples were within the normal range at which most freshwater fish species become non-stressed. Of a total of 674 samples included in the study, bacteria were isolated from 154(22.8%) samples with significant difference (P < 0.05) observed in some isolates with respect to sample origin. Of these 154 isolates, 103(66.8%) isolates were gram-negative bacteria consisting of 15 species based on morphology and a range of biochemical tests. From live fish samples, Escherichia coli was the dominant species with 15 isolates followed by Edwardsiella tarda (12), Salmonella Paratyphi (10), Salmonella Typhi (9), Shigella dysenteriae (7), Shigella flexneri (7), Klebsiella pneumonia (7), Enterobacter aerogenes (6), Enterobacter cloacae (5), Pseudomonas aeruginosa (5), Vibrio parahemolyticus (5), Aeromonas sobria (4), Citrobacter freundii (4), Citrobacter koseri (4) and Plesiomonas shigelloides (3). The detection of the common fecal coliforms (E. coli, K. pneumoniae and E. aerogenes) and Salmonella spp. in processed fish indicates the potential danger of passage of pathogenic bacteria and/or their poisons to humans via infected and/or contaminated fish products. Human infection by pathogenic fish bacteria and food poisoning is possible through contamination of fish product in fish production chain due to inadequate handling, poor hygiene and contact with contaminated water. Therefore, producers, consumers and all other stakeholders need to be cautious during handling, processing and consumption of fish harvested from the study lakes.
Topics: Animals; Bacteria; Carps; Cross-Sectional Studies; Escherichia coli; Fish Products; Gram-Negative Bacteria; Lakes; Water; Ethiopia
PubMed: 36517783
DOI: 10.1186/s12917-022-03508-w -
Microbiology Resource Announcements Jan 2023Here, we present a draft genome sequence of Plesiomonas shigelloides MD22D9, isolated from the digestive tract of the North American medicinal leech Macrobdella decora....
Here, we present a draft genome sequence of Plesiomonas shigelloides MD22D9, isolated from the digestive tract of the North American medicinal leech Macrobdella decora. The gut microbiome of the medicinal leech is hypothesized to be critical for maintaining host fitness. This genome can provide insights into this uncharacterized microbe-host relationship.
PubMed: 36515507
DOI: 10.1128/mra.00939-22 -
BMC Microbiology Dec 2022RpoN, also known as σ, first reported in Escherichia coli, is a subunit of RNA polymerase that strictly controls the expression of different genes by identifying...
BACKGROUND
RpoN, also known as σ, first reported in Escherichia coli, is a subunit of RNA polymerase that strictly controls the expression of different genes by identifying specific promoter elements. RpoN has an important regulatory function in carbon and nitrogen metabolism and participates in the regulation of flagellar synthesis, bacterial motility and virulence. However, little is known about the effect of RpoN in Plesiomonas shigelloides.
RESULTS
To identify pathways controlled by RpoN, RNA sequencing (RNA-Seq) of the WT and the rpoN deletion strain was carried out for comparison. The RNA-seq results showed that RpoN regulates ~ 13.2% of the P. shigelloides transcriptome, involves amino acid transport and metabolism, glycerophospholipid metabolism, pantothenate and CoA biosynthesis, ribosome biosynthesis, flagellar assembly and bacterial secretion system. Furthermore, we verified the results of RNA-seq using quantitative real-time reverse transcription PCR, which indicated that the absence of rpoN caused downregulation of more than half of the polar and lateral flagella genes in P. shigelloides, and the ΔrpoN mutant was also non-motile and lacked flagella. In the present study, the ability of the ΔrpoN mutant to kill E. coli MG1655 was reduced by 54.6% compared with that of the WT, which was consistent with results in RNA-seq, which showed that the type II secretion system (T2SS-2) genes and the type VI secretion system (T6SS) genes were repressed. By contrast, the expression of type III secretion system genes was largely unchanged in the ΔrpoN mutant transcriptome and the ability of the ΔrpoN mutant to infect Caco-2 cells was also not significantly different compared with the WT.
CONCLUSIONS
We showed that RpoN is required for the motility and contributes to the killing ability of P. shigelloides and positively regulates the T6SS and T2SS-2 genes.
Topics: Humans; RNA Polymerase Sigma 54; Gene Expression Regulation, Bacterial; Plesiomonas; Bacterial Proteins; Escherichia coli; Caco-2 Cells
PubMed: 36510135
DOI: 10.1186/s12866-022-02722-8 -
The American Journal of Tropical... Jan 2023Diarrhea is a common cause of morbidity and mortality among children younger than 5 years in developing countries. Children from 3 to 60 months of age were recruited...
Diarrhea is a common cause of morbidity and mortality among children younger than 5 years in developing countries. Children from 3 to 60 months of age were recruited from two hospitals in Nepal- Bharatpur Hospital, Bharatpur, and Kanti Children's Hospital, Kathmandu-in 2006 to 2009. Stool specimens collected from 1,200 children with acute diarrhea (cases) and 1,200 children without diarrhea (control subjects) were examined for a broad range of enteropathogens by standard microbiology, including microscopy, enzyme immunoassay for viral pathogens (adenovirus, astrovirus, and rotavirus) and protozoa (Giardia, Cryptosporidium, and Entamoeba histolytica), as well as by using reverse transcription real-time polymerase for norovirus. Antimicrobial susceptibility testing was performed using the disk diffusion method. Overall, rotavirus (22% versus 2%), norovirus (13% versus 7%), adenovirus (3% versus 0%), Shigella (6% versus 1%), enterotoxigenic Escherichia coli (8% versus 4%), Vibrio (7% versus 0%), and Aeromonas (9% versus 3%) were identified significantly more frequently in cases than control subjects. Campylobacter, Plesiomonas, Salmonella, and diarrheagenic E. coli (enteropathogenic, enteroinvasive, enteroaggregative) were identified in similar proportions in diarrheal and non-diarrheal stools. Campylobacter was resistant to second-generation quinolone drugs (ciprofloxacin and norfloxacin), whereas Vibrio and Shigella were resistant to nalidixic acid and trimethoprim/sulfamethoxazole. This study documents the important role of rotavirus and norovirus in acute diarrhea in children younger than 5 years, followed by the bacteria Shigella, enterotoxigenic E. coli, Vibrio cholera, and Aeromonas. Data on the prevalence and epidemiology of enteropathogens identify potential pathogens for public health interventions, whereas pathogen antibiotic resistance pattern data may provide guidance on choice of therapy in clinical settings.
Topics: Humans; Infant; Child, Preschool; Cryptosporidiosis; Nepal; Cryptosporidium; Diarrhea; Rotavirus; Shigella; Enterotoxigenic Escherichia coli; Campylobacter; Adenoviridae; Norovirus; Adenoviridae Infections; Anti-Infective Agents; Acute Disease
PubMed: 36509064
DOI: 10.4269/ajtmh.21-1219 -
Frontiers in Cellular and Infection... 2022The influence of dietary probiotic on the growth performance, digestive enzyme activity, immune parameters and disease resistance of yellow catfish () was evaluated....
The influence of dietary probiotic on the growth performance, digestive enzyme activity, immune parameters and disease resistance of yellow catfish () was evaluated. Commercial diet (C) or diet containing 10 cfu/g (T) was fed for 4 weeks, and final weight (FW), specific growth rate (SGR) and feed conversion ratio (FCR) were improved (0.05) in the T group. Dietary increased protease and amylase activities in the digestive tract after 2 and 4 weeks, respectively. Respiratory burst (RB), plasma lysozyme (LZM) activity, total antioxidant capacity (T-AOC) and superoxide dismutase (SOD) activity were also elevated (0.05). Immune-related genes signal transducer and activator of transcription 1 (STATA-1), immunoglobulin M (IgM) and C-type lectin (CTL) were upregulated (0.05), but interleukin-1 β (IL-1β) was not (0.05). Intestinal microbiota analysis showed that the community structure was significantly different between the two groups; the relative abundance of was increased but was decreased in T. Moreover, challenge tests showed that the resistance of fish fed against and was significantly enhanced (0.05). In conclusion, dietary supplementation of can effectively improve the growth performance, digestive enzyme activity, immune responses, intestinal microbiota composition and disease resistance of yellow catfish.
Topics: Animals; Catfishes; Bacillus amyloliquefaciens; Gastrointestinal Microbiome; Disease Resistance; Diet; Gastrointestinal Agents
PubMed: 36452300
DOI: 10.3389/fcimb.2022.1047351 -
Vaccines Nov 2022The swift emergence of antibiotic resistance (AR) in bacterial pathogens to make themselves adaptable to changing environments has become an alarming health issue. To...
The swift emergence of antibiotic resistance (AR) in bacterial pathogens to make themselves adaptable to changing environments has become an alarming health issue. To prevent AR infection, many ways can be accomplished such as by decreasing the misuse of antibiotics in human and animal medicine. Among these AR bacterial species, is one of the etiological agents of intestinal infection in humans. It is a gram-negative rod-shaped bacterium that is highly resistant to several classes of antibiotics, and no licensed vaccine against the aforementioned pathogen is available. Hence, substantial efforts are required to screen protective antigens from the pathogen whole genome that can be subjected easily to experimental evaluations. Here, we employed a reverse vaccinology (RV) approach to design a multi-antigenic epitopes based vaccine against . The complete genomes of were retrieved from the National Center for Biotechnological Information (NCBI) that on average consist of 5226 proteins. The complete proteomes were subjected to different subtractive proteomics filters, and in the results of that analysis, out of total proteins, 2399 were revealed as non-redundant and 2827 as redundant proteins. The non-redundant proteins were further checked for subcellular localization analysis, in which three were localized in the extracellular matrix, eight were outer membrane, and 13 were found in the periplasmic membrane. All surface localized proteins were found to be virulent. Out of a total of 24 virulent proteins, three proteins (flagellar hook protein (FlgE), hypothetical protein, and TonB-dependent hemoglobin/transferrin/lactoferrin family receptor protein) were considered as potential vaccine targets and subjected to epitopes prediction. The predicted epitopes were further examined for antigenicity, toxicity, and solubility. A total of 10 epitopes were selected (GFKESRAEF, VQVPTEAGQ, KINENGVVV, ENKALSQET, QGYASANDE, RLNPTDSRW, TLDYRLNPT, RVTKKQSDK, GEREGKNRP, RDKKTNQPL). The selected epitopes were linked with each other via specific GPGPG linkers in order to design a multi-epitopes vaccine construct, and linked with cholera toxin B subunit adjuvant to make the designed vaccine construct more efficient in terms of antigenicity. The 3D structure of the vaccine construct was modeled ab initio as no appropriate template was available. Furthermore, molecular docking was carried out to check the interaction affinity of the designed vaccine with major histocompatibility complex (MHC-)I (PDB ID: 1L1Y), MHC-II (1KG0), and toll-like receptor 4 ((TLR-4) (PDB: 4G8A). Molecular dynamic simulation was applied to evaluate the dynamic behavior of vaccine-receptor complexes. Lastly, the binding free energies of the vaccine with receptors were estimated by using MMPB/GBSA methods. All of the aforementioned analyses concluded that the designed vaccine molecule as a good candidate to be used in experimental studies to disclose its immune protective efficacy in animal models.
PubMed: 36366394
DOI: 10.3390/vaccines10111886 -
Antioxidants (Basel, Switzerland) Oct 2022Flavonoids have antimicrobial and anti-oxidation properties. The effects of the flavonoid quercetin on the intestinal microflora of freshwater dark sleeper were tested...
Flavonoids have antimicrobial and anti-oxidation properties. The effects of the flavonoid quercetin on the intestinal microflora of freshwater dark sleeper were tested for the first time. juveniles were treated with quercetin for 21 days at one of three concentrations (2.5, 5.0, or 10.0 mg/L) and compared with a control group that was not treated with quercetin. Quercetin improved the stability of the intestinal flora in and the probiotic bacteria spp. and spp. increased in species abundance after the low concentration quercetin treatments. Furthermore, the abundance of pathogenic bacteria spp., spp., and spp. decreased after the fish had been exposed to quercetin. Activity of hepatic antioxidant enzymes (superoxide dismutase, SOD), (glutathione S-transferase, GST), (glutathione peroxidase, GSH-Px), and (total antioxidant capacity, T-AOC) increased in the livers of treated with quercetin, thereby increasing their hepatic antioxidant capacity and their ability to scavenge free radicals.
PubMed: 36290739
DOI: 10.3390/antiox11102015 -
Animals : An Open Access Journal From... Oct 2022This study was conducted to investigate the effects of meal (TM) replacement for fish meal (FM) on growth performance, humoral immunity, and intestinal health of...
This study was conducted to investigate the effects of meal (TM) replacement for fish meal (FM) on growth performance, humoral immunity, and intestinal health of juvenile large yellow croakers (). Four experimental diets were formulated by replacing FM with TM at different levels-0% (TM0), 15% (TM15), 30% (TM30), and 45% (TM45). Triplicate groups of juveniles (initial weight = 11.80 ± 0.02 g) were fed the test diets to apparent satiation two times daily for eight weeks. There was no significant difference in final body weight (FBW) and weight gain rate (WG) among TM0, TM15, and TM30, while TM45 feeding significantly reduced the FBW and WG. Compared with TM0, AKP activity in serum was significantly decreased in TM45, while the TM15 group remarkably increased LZM activity. TM30 showed significantly higher serum C3 levels compared to the TM0 group, while the TM addition groups decreased the C4 levels significantly in the serum. In terms of intestinal histology, the addition of TM increased the height and thickness of the intestinal villus and also increased the thickness of the intestinal muscles significantly. The addition of TM significantly reduced the serum DAO and D-lactate concentrations. The results of 16S rRNA gene sequencing showed that the addition of TM significantly enhanced the relative abundance of Bacilli and and contributed to the decrease in the relative abundance of . In addition, the TM30 and TM45 groups significantly reduced the abundance of Peptostreptococcaceae. Overall, our results indicated that TM could be a viable alternative protein source, 6.7% TM supplantation (replacing 15% FM) in large yellow croaker feed improved humoral immunity and intestinal health with no adverse effects on growth. Furthermore, the replacement of FM with 30% and 45% TM adversely affects growth and humoral immunity.
PubMed: 36230400
DOI: 10.3390/ani12192659 -
Microbiology Spectrum Oct 2022Maintaining the health of seafarers is a difficult task during long-term voyages. Little is known about the corresponding changes in the gut microbiome-host interaction....
Maintaining the health of seafarers is a difficult task during long-term voyages. Little is known about the corresponding changes in the gut microbiome-host interaction. This study recruited 30 seafarers undertaking a 6-month voyage and analyzed their gut microbiota using 16S rRNA gene sequencing. Fecal untargeted metabolomics analysis was performed using liquid chromatography-mass spectrometry. Significant changes in the composition of the gut microbiota and an increased ratio of / at the end (day 180) of the 6-month voyage, relative to the start (day 0), were observed. At the genus level, the abundances of and were significantly increased, while the abundance of was decreased. Predicted microbial functional analysis revealed significant decreases in folate biosynthesis and biotin metabolism. Furthermore, 20 differential metabolites within six differentially enriched human metabolic pathways (including arginine biosynthesis, lysine degradation, phenylalanine metabolism, sphingolipid metabolism, pentose and glucuronate interconversions, and glycine, serine, and threonine metabolism) were identified by comparing the fecal metabolites at day 0 and day 180. Spearman correlation analysis revealed close relationships between the 14 differential microbiota members and the six differential fecal metabolites that might affect specific human metabolic pathways. This study adopted a multi-omics approach and provides potential targets for maintaining the health of seafarers during long-term voyages. These findings are worthy of more in-depth exploration in future studies. Maintaining the health of seafarers undertaking long-term voyages is a difficult task. Apart from the alterations in the gut microbiome and fecal metabolites after a long-term voyage, our study also revealed that 20 differential metabolites within six differentially enriched human metabolic pathways are worthy of attention. Moreover, we found close relationships between the 14 differential microbiota members and the six differential fecal metabolites that might impact specific human metabolic pathways. Accordingly, preventative measures, such as adjusting the gut microbiota by decreasing potential pathobionts or increasing potential probiotics as well as offsetting the decrease in B vitamins and beneficial metabolites (e.g., d-glucuronic acid and citrulline) via dietary adjustment or nutritional supplements, might improve the health of seafarers during long-term sea voyages. These findings provide valuable clues about gut microbiome-host interactions and propose potential targets for maintaining the health of seafarers engaged in long-term sea voyages.
Topics: Humans; Gastrointestinal Microbiome; RNA, Ribosomal, 16S; Vitamin B Complex; Citrulline; Biotin; Lysine; Metabolomics; Feces; Pentoses; Glucuronates; Glycine; Glucuronic Acid; Serine; Phenylalanine; Sphingolipids; Threonine; Arginine; Folic Acid
PubMed: 36197290
DOI: 10.1128/spectrum.01899-22 -
International Journal of Biological... Dec 2022A 56-day feeding trial was conducted to assess the effects of different viscous guar gum on the growth, intestinal flora, and intestinal health of Micropterus salmoides....
A 56-day feeding trial was conducted to assess the effects of different viscous guar gum on the growth, intestinal flora, and intestinal health of Micropterus salmoides. Four practical diets with 42.5 % crude protein and 13.7 % crude lipid were formulated to contain 8 % cellulose and three different viscosities (2500, 5200, and 6000 mPa·s) of guar gum. Dietary guar gum inhibits fish growth and feed utilization, decreases the α-diversity of the intestinal flora, and negatively alters the intestinal flora structure and metabolite composition. High viscous guar gum down-regulated the intestinal tight junction, anti-inflammatory, and anti-apoptotic related gene's expression, decreased digesta butyrate/histamine ratio; and increased the abundance of Plesiomonas shigelloides. These results suggest that dietary guar gum adversely affects intestinal health by disrupting intestinal flora structure and metabolite composition, and that viscosity should be considered when using guar gum as a binder in aquafeeds.
Topics: Animals; Viscosity; Bass; Gastrointestinal Microbiome; Galactans; Plant Gums; Mannans; Dietary Fiber
PubMed: 36181882
DOI: 10.1016/j.ijbiomac.2022.09.220