scholarly journals A First Insight into the Structural and Functional Comparison of Environmental Microbiota in Freshwater Turtle Chinemys reevesii at Different Growth Stages under Pond and Greenhouse Cultivation

2020 ◽  
Vol 8 (9) ◽  
pp. 1277 ◽  
Author(s):  
Aiguo Zhou ◽  
Shaolin Xie ◽  
Di Sun ◽  
Pan Zhang ◽  
Han Dong ◽  
...  

The microbial community structure of water is an important indicator for evaluating the water quality of the aquaculture environment. In this study, the investigation and comparison of the bacterial communities of pond cultivation (PC) and greenhouse cultivation (GC) between hatchling, juvenile, and adult growth stages of C. reevesii were performed. In addition, the V4 regions of the 16S rRNA gene were sequenced. The Chao1 richness estimator of the PC group was significantly higher than that of the GC group. The beta diversity showed that the microbiotas of the two groups were isolated from each other. The dominant phyla were Cyanobacteria, Proteobacteria, Actinobacteria, Bacteroidetes, Verrucomicrobia, and Planctomycetes in the PC group and Proteobacteria, Bacteroidetes, Firmicutes, Cyanobacteria, Chloroflexi, and Actinobacteria in the GC group. Both the numbers and the types of Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway annotations differed between the PC and GC groups. The prediction of bacterial phenotype implied that the GC environment is more likely to deteriorate, and turtles are more susceptible to pathogens than those of the PC environment. In addition, a total of nine potential pathogenic bacteria were identified and the correlation of environmental factors analyses showed significant differences of bacterial species between the PC and GC groups, while the potential pathogenic bacteria showed significant correlation with the stocking density, temperature, pH, orthophosphate (PO4-P), and dissolved oxygen (DO) in both the PC and GC groups. Noticeably, this is the first report to describe the different microbiota characteristics of the different cultivation environments in the different growth stages of C. reevesii, which will provide valuable data for water quality adjustment, disease prevention, and the healthy breeding of turtles.

2020 ◽  
Author(s):  
Aiguo Zhou ◽  
Shaolin Xie ◽  
Di Sun ◽  
Pan Zhang ◽  
Zhengkun Pan ◽  
...  

Abstract The microbial community structure is an important indicator for evaluating the water quality of the aquaculture environment. In this study, V4 regions of 16S rRNA gene of pond (PC) and greenhouse cultured (GC) C. reevesii were sequenced. Results showed that a total of 1,993,090 high quality counts and 105,159 observed OUTs were obtained; and the Chao1 richness estimator of PC was significantly higher than that of GC groups. Beta-diversity showed that the microbiota of two groups were isolated from each other. In addition, the correlation analysis of environmental factors showed that NO2-N, PH, PO4-P, and stocking density played significant roles in the bacterial community composition. The dominant phyla in PC groups were cyanobacteria, proteobacteria, actinobacteria, bacteroidetes, verrucomicrobia, planctomycetes; and in GC groups were proteobacteria, bacteroidetes, firmicutes, cyanobacteria, chloroflexi, actinobacteria. The functional prediction showed that the top5 Picrust prediction gene functions were protein processing in endoplasmic reticulum, retinol metabolism, proteasome, glycan binding proteins, and stilbenoid, diarylheptanoid and gingerol biosynthesis. Meanwhile, the numbers and types of KEGG pathway annotations showed a significant difference between the two cultivation environments. The prediction of bacterial phenotype implied that the GC environment is more likely to deteriorate, and turtles are more susceptible to pathogens than those of PC environment. This is the first report to explore and understand the difference of microbiota characteristics between different cultivation environments in different growth stages of C. reevesii, which will provide basic data for water quality adjustment, disease prevention, and healthy breeding of turtle.


2019 ◽  
Vol 97 (Supplement_3) ◽  
pp. 242-243
Author(s):  
Xiaofan Wang ◽  
Xiaoyuan Wei ◽  
Feilong Deng ◽  
Tsungcheng Tsai ◽  
Charles V Maxwell ◽  
...  

Abstract Substantial progress has been made in the culture-omics of the human gut microbiota. However, little is known about the culture-omics of the swine gut microbiota, despite recent reports of their significant roles in swine health and production. To fill this knowledge gap in research, we tested 52 bacterial cultivation methods with different media and gas combinations. Fresh fecal samples (0.2g/sample) were collected from three pigs at the end of four growth stages: lactation, nursery, growing and finishing and were mixed with a stomacher in 20 mL saline. Aliquots of 50 uL microbial suspensions were then spread onto different media plates and incubated under aerobic and anaerobic conditions at 37C for up to 5 days. An additional aliquot of each sample was subjected to direct DNA extraction as a positive control. Bacterial colonies from each plate were collected and DNA was extracted from these samples using the Powersoil DNA isolation kit and sequenced with an Illumina Miseq sequencer targeting the V4 region of the 16S rRNA gene. Sequences were analyzed with the Deblur algorithm in the QIIME2 package. A total of 378, 482, 565, and 555 bacterial features were observed from microbial solutions at the end of lactation, nursery, growing and finishing. Our culturing methods recovered 415, 675, 808, and 823 features correspondingly, representing 45.2%, 54.8%, 53.3%, and 56.4% of total features observed in microbial solutions. The top ten most easily cultured genus were Escherichia, Streptococcus, Lactobacillus, Megasphaera, Acidaminococcus, Bacillus, Mitsuokella, Enterococcus and Prevotella. Non-parametric permutational multivariate analysis of variance shows that the main factors driving the swine culture-omics included medium, age and oxygen condition. This study identifies the cultivable bacteria from fecal samples collected at different growth stages of pigs and provides a guidance to cultivate potential beneficial or pathogenic bacteria of interests and validate their functions in swine production.


2014 ◽  
Vol 2014 ◽  
pp. 1-11 ◽  
Author(s):  
K. Böhme ◽  
P. Cremonesi ◽  
M. Severgnini ◽  
Tomás G. Villa ◽  
I. C. Fernández-No ◽  
...  

Traditional culturing methods are still commonly applied for bacterial identification in the food control sector, despite being time and labor intensive. Microarray technologies represent an interesting alternative. However, they require higher costs and technical expertise, making them still inappropriate for microbial routine analysis. The present study describes the development of an efficient method for bacterial identification based on flow-through reverse dot-blot (FT-RDB) hybridization on membranes, coupled to the high specific ligation detection reaction (LDR). First, the methodology was optimized by testing different types of ligase enzymes, labeling, and membranes. Furthermore, specific oligonucleotide probes were designed based on the 16S rRNA gene, using the bioinformatic tool Oligonucleotide Retrieving for Molecular Applications (ORMA). Four probes were selected and synthesized, being specific forAeromonasspp.,Pseudomonasspp.,Shewanellaspp., andMorganella morganii, respectively. For the validation of the probes, 16 reference strains from type culture collections were tested by LDR and FT-RDB hybridization using universal arrays spotted onto membranes. In conclusion, the described methodology could be applied for the rapid, accurate, and cost-effective identification of bacterial species, exhibiting special relevance in food safety and quality.


2018 ◽  
Author(s):  
Zongfu Hu ◽  
Xi CHEN ◽  
Jie CHANG ◽  
Jianhua YU ◽  
Qing TONG ◽  
...  

Widely distributed across the world, the freshwater snail Radix auricularia plays an important role in freshwater systems. In this study, gut bacterial communities of R. auricularia were characterized using 16S rRNA amplicon sequencing, then intestinal bacteria were compared at different growth stages: adult snails (AS) (with complete gonadal development) and juvenile snails (JS) (with incomplete gonadal development). We obtained 251,072 high quality sequences which were clustered into 1,196 operational taxonomic units (OTUs) with 97% sequence identity. The predominant phyla were Proteobacteria and Cyanobacteria, followed by Chloroflexi, Firmicutes, and Actinobacteria. Other bacterial species such as Tenericutes, Bacteroidetes, Fusobacteria and Verrucomicrobia were present to a lesser extent. 52 bacterial families and 55 genera were found in > 1% of each sample. A large number of species could not be successfully identified. 469 core OTUs were found to make up 39.38% of all OTUs and 88.38% of all sequences. Samples obtained from juvenile organisms possessed higher ratios of Ruminococcaceae, Subdoligranulum, and Faecalibacterium than adult species. Furthermore, 16S rRNA gene data was used to predict function, showing that genes related to metabolism and environmental information processing were rich in snail samples.


2021 ◽  
Vol 99 (Supplement_3) ◽  
pp. 135-136
Author(s):  
Rebecca K Poole

Abstract Over the past decade, a multitude of research has sought to understand the complexity and role of the reproductive microbiome as it pertains to fertility. Previously, the reproductive microbiome was evaluated using culture-dependent methods; however, recent advancements in culture-independent, 16S rRNA gene amplicon community sequencing have vastly expanded our understanding of the reproductive tract microbiome. Early sequencing studies sought to compare the vaginal microbiome of cattle to the vaginal microbiome of healthy women, which predominantly consists of bacteria in the genus Lactobacillus and believed to be an indicator of fertility. In the vagina of beef cattle, however, there are incredibly low abundances of Lactobacillus and a greater diversity of bacterial species present. Beta-diversity, which examine differences in bacterial communities between samples, does not appear to differ in the vagina between unbred, open, or pregnant cattle. In postpartum beef cattle just prior to breeding, there are greater levels of diversity and increased bacterial species richness in the vagina compared to the uterus. Research on bacterial species within the uterus have primarily focused on pathogenic bacteria in postpartum cattle diagnosed with uterine disease. Fewer studies have investigated uterine bacterial species in presumed healthy postpartum beef cattle and the subsequent effects on fertility outcomes (e.g., pregnant vs. open at day 30). When evaluating the uterine microbiome during an industry standard estrus synchronization protocol, bacterial community abundance and diversity reduce over time regardless of resulting fertility outcomes. The greatest difference in uterine bacterial abundance between resulting pregnant and non-pregnant cattle appears to occur just prior to breeding. Numerous mechanisms could be contributing to the fluctuations in the uterine microbiome in beef cattle including circulating hormone concentrations or local immunoregulation. This presentation will focus on recent research investigating potential mechanisms that may alter the reproductive microbiome and ultimately impact fertility.


2020 ◽  
Vol 5 (2) ◽  
pp. 93-102
Author(s):  
Soumya Chatterjee ◽  
Sibnarayan Datta ◽  
Leeza Banu ◽  
Mohan G. Vairale ◽  
Sonika Sharma

Microflora plays an important role in modulating environmental quality. Among microflora, bacteria are omnipresent in the environment. Pathogenic bacteria, present in air, are known to affect significantly the health and well-being of human, animal or plant populations. Air bacteria monitoring is thus essential for surveillance of pathogenic microorganisms from public health perspective besides its significant implications in detection and mitigation of biothreat related issues. Despite the geo-politically strategic importance of northeast India, there is scarcity of data on human health and disease surveillance. Considering these facts, we, for the first time studied the bacterial diversity of air at six important sites adjacent to the international border in the northeast region of India, having an altitude range of 73 m (Tezpur) to 4170 m (Sela Pass) above sea level. Standard microbiological techniques, such as Tryptone Soya Agar, Mannitol salt and McConkey agar strips and plates were used for air bacterial load assessment and culture for subsequent analysis using biochemical and molecular techniques. Following RFLP study, twenty six different bacterial colonies were isolated. Subsequently, bacteria identification was carried out by examining the substrate utilisation patterns, sequencing 16S rRNA gene and phylogenetic analysis. Results of the study reveal that the isolates mostly belong to two genera Bacillus and Staphylococcus (eleven in each genus), along with Micrococcus, Pseduomonas and Acinetobacter. Based on significant match of our sequences with that of medically important bacterial 16S rRNA sequences available at 16SpathDB 2.0 and review of available literature, we found that a number of these bacterial species have the pathogenic potential. In this manuscript we report our results and discuss the importance of air bacterial surveillance from the perspective of human health, hygiene and biothreat mitigation.


2021 ◽  
Vol 15 (Supplement_1) ◽  
pp. S594-S595
Author(s):  
S Ellul ◽  
P Rausch ◽  
A Pisani ◽  
C Bang ◽  
P Ellul ◽  
...  

Abstract Background The role of microbiome with the alteration between commensal and pathogenic bacteria, has been linked to IBD. Meanwhile Escherichia coli Nissle 1917, Lactobacillus rhamnosus GG (LGG) and faecal transplantation are used in IBD. The aim of this study was to prospectively determine faecal microbiota composition of newly diagnosed treatment naïve IBD patients. Methods Patients diagnosed with IBD between January 2018-September 2019 were recruited. Clinical data was collected and patients asked to submit stool samples for microbiome analysis. Stool samples from a control population were recruited and analysed via the bacterial 16s rRNA gene sequencing on illumine MiSeq. Results 100 IBD patients (CD: n=46, UC: n=53 & IBDU: n=1) and 97 controls with specific inclusion and exclusion criteria collected. IBD patients were noted to display reduced average species richness and community evenness compared to healthy controls (Alpha- Diversity) (Figure 1). Beta-diversity between microbial communities of healthy individuals and IBD patients was significantly different, but no observed separation between the two types of IBD was noted (Figure 2). 11 ASVs were abundant in CD patients including: ASV-70 – Lactobacillus gasseri, Klebsiella uncl., Candidatus-saccharibacteria, ASV-157 - Acteroides clarus and ASV 249- Parasutterella uncl. In UC cohort, 10 ASVs were abundant including: ASV 6-Escherichia/Shigella uncl., ASB-41-Sutterella wadsworthensis, ASV 44- Bacteroides faecis and Actinobacteria. An association between UC and ASV 313 (Faecalibacteria) was present. In the microbiome of healthy controls, 20 ASVs were abundant, including: ASV-14 G-Alistipes uncl., ASV 20-(Akkermansia muciniphila),(bacterium belonging to the phylum Verrucomicrobia), ASV 321 (Clostridia uncl.), ASV 96 (Rumminococcaceae uncl.), Alistepes uncl. (ASV 61), Subdoligranulum uncl. (ASV 453) and the unclassifiable bacteria. A higher amount of Verrucomicrobia was present in the healthy group as opposed to the IBD. Conclusion ASV 249- Parasutterella unlc., was indicative of CD associated microbiome through the indicator species analysis. Typical microbiome changes in IBD patients include increased abundance of the pro-inflammatory species with a reduction in anti-inflammatory bacterial species, with a noticeable reduction in alpha and beta diversity. In the local cohort, a particular change in the local α- and β diversity was noted to be present between healthy controls and IBD cohort. This could be a potential way in which targeted therapeutic approaches using specific dosage and durations of probiotic or faecal transplant can be used to alter faecal microbiome using specific bacteria present in healthy controls and with elimination of potentially harmful bacteria in IBD patients. Figure 1: Alpha diversity between different Groups using Chao1 species richness and Simpson 1-D Figure 2: Beta diversity between different groups using Bray-Curtis dissimilarity, Jaccard distance.


2013 ◽  
Vol 8 (11) ◽  
pp. 1934578X1300801 ◽  
Author(s):  
Roberto Cecotti ◽  
Emanuele Carpana ◽  
Patrizia Bergomi ◽  
Aldo Tava

The composition of the volatile fraction of Trifolium pratense L. ssp. nivale (Sieber) Asch. et Gr. from the southwestern Alps was investigated. Fresh aerial parts were collected in the summer at three different growth stages, namely vegetative, flowering and fruiting. The oils obtained by steam-distillation accounted for 0.006 to 0.011% of the fresh plant material and their composition was determined by GC/FID and GC/MS. Several classes of compounds were found, with a predominance of alcohols in all phases, followed by aldehydes, hydrocarbons, terpenes, phenolics, ketones, acids and esters. The oil composition varied both in quantity and quality; the most abundant compounds were oct-1-en-3-ol and phenylacetaldehyde in the vegetative and flowering phases, and phenylacetaldehyde and 2-phenylethanol in the fruiting phase. The essential oils obtained were tested for activity against two major bee pests, i.e. Paenibacillus larvae and Melissococcus plutonius, and against a reference bacterial species, Bacillus subtilis.


2018 ◽  
Author(s):  
Zongfu Hu ◽  
Xi CHEN ◽  
Jie CHANG ◽  
Jianhua YU ◽  
Qing TONG ◽  
...  

Widely distributed across the world, the freshwater snail Radix auricularia plays an important role in freshwater systems. In this study, gut bacterial communities of R. auricularia were characterized using 16S rRNA amplicon sequencing, then intestinal bacteria were compared at different growth stages: adult snails (AS) (with complete gonadal development) and juvenile snails (JS) (with incomplete gonadal development). We obtained 251,072 high quality sequences which were clustered into 1,196 operational taxonomic units (OTUs) with 97% sequence identity. The predominant phyla were Proteobacteria and Cyanobacteria, followed by Chloroflexi, Firmicutes, and Actinobacteria. Other bacterial species such as Tenericutes, Bacteroidetes, Fusobacteria and Verrucomicrobia were present to a lesser extent. 52 bacterial families and 55 genera were found in > 1% of each sample. A large number of species could not be successfully identified. 469 core OTUs were found to make up 39.38% of all OTUs and 88.38% of all sequences. Samples obtained from juvenile organisms possessed higher ratios of Ruminococcaceae, Subdoligranulum, and Faecalibacterium than adult species. Furthermore, 16S rRNA gene data was used to predict function, showing that genes related to metabolism and environmental information processing were rich in snail samples.


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