Host Reproductive Cycle Influences the Pouch Microbiota of Wild Southern Hairy-Nosed Wombats (Lasiorhinus Latifrons)

Abstract Background: Marsupials are born much earlier than placental mammals, with most crawling from the birth canal to the protective marsupium (pouch) to further their development. However, little is known about the microbiology of the pouch and how it changes throughout a marsupial’s reproductive cycle. Here, using stringent controls, we characterized the microbial composition of multiple body sites from 26 wild Southern Hairy-nosed Wombats (SHNWs), including pouch samples from animals at different reproductive stages. Results: Using qPCR of the 16S rRNA gene we found higher concentrations of microbial DNA in the pouch than in negative controls. We observed significant differences in microbial composition and diversity between the body sites tested, as well as between pouch samples from different reproductive stages. The pouches of reproductively active females had drastically lower microbial diversity (mean richness 19) compared to reproductively inactive females (mean richness 941), and were dominated by gram positive bacteria from the Actinobacteriota phylum (81.7-90.6%), with the dominant families classified as Brevibacteriaceae, Corynebacteriaceae, Microbacteriaceae, and Dietziaceae. Three of the five most abundant sequences identified in reproductively active pouches had closest matches to microbes previously isolated from tammar wallaby pouches. Conclusions: This study represents the first contamination controlled investigation into the marsupial pouch microbiota, and sets a rigorous framework for future pouch microbiota studies. Our results suggest that SHNW pouches contain communities of microorganisms that are substantially altered by the host reproductive cycle. We recommend further investigation into the roles that pouch microorganisms may play in marsupial reproductive health and joey survival.

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Host reproductive cycle influences the pouch microbiota of wild southern hairy-nosed wombats (Lasiorhinus latifrons)

Animal Microbiome ◽

10.1186/s42523-021-00074-8 ◽

2021 ◽

Vol 3 (1) ◽

Author(s):

Sesilje Weiss ◽

David Taggart ◽

Ian Smith ◽

Kristofer M. Helgen ◽

Raphael Eisenhofer

Keyword(s):

Microbial Community ◽

Reproductive Cycle ◽

Tammar Wallaby ◽

The Body ◽

Rrna Gene ◽

Microbial Composition ◽

Birth Canal ◽

Gram Positive Bacteria ◽

Rigorous Framework ◽

The 16S Rrna Gene

Abstract Background Marsupials are born much earlier than placental mammals, with most crawling from the birth canal to the protective marsupium (pouch) to further their development. However, little is known about the microbiology of the pouch and how it changes throughout a marsupial’s reproductive cycle. Here, using stringent controls, we characterized the microbial composition of multiple body sites from 26 wild Southern Hairy-nosed Wombats (SHNWs), including pouch samples from animals at different reproductive stages. Results Using qPCR of the 16S rRNA gene we detected a microbial community in the SHNW pouch. We observed significant differences in microbial composition and diversity between the body sites tested, as well as between pouch samples from different reproductive stages. The pouches of reproductively active females had drastically lower microbial diversity (mean ASV richness 19 ± 8) compared to reproductively inactive females (mean ASV richness 941 ± 393) and were dominated by gram positive bacteria from the Actinobacteriota phylum (81.7–90.6%), with the dominant families classified as Brevibacteriaceae, Corynebacteriaceae, Microbacteriaceae, and Dietziaceae. Three of the five most abundant sequences identified in reproductively active pouches had closest matches to microbes previously isolated from tammar wallaby pouches. Conclusions This study represents the first contamination-controlled investigation into the marsupial pouch microbiota, and sets a rigorous framework for future pouch microbiota studies. Our results indicate that SHNW pouches contain communities of microorganisms that are substantially altered by the host reproductive cycle. We recommend further investigation into the roles that pouch microorganisms may play in marsupial reproductive health and joey survival.

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Host reproductive cycle influences the pouch microbiota of wild Southern Hairy-nosed Wombats (Lasiorhinus latifrons)

10.21203/rs.3.rs-28743/v3 ◽

2021 ◽

Author(s):

Sesilje Weiss ◽

David Taggart ◽

Ian Smith ◽

Kristofer M Helgen ◽

Raphael Eisenhofer

Keyword(s):

Microbial Community ◽

Reproductive Cycle ◽

Tammar Wallaby ◽

The Body ◽

Rrna Gene ◽

Microbial Composition ◽

Birth Canal ◽

Gram Positive Bacteria ◽

Rigorous Framework ◽

The 16S Rrna Gene

Abstract Background Marsupials are born much earlier than placental mammals, with most crawling from the birth canal to the protective marsupium (pouch) to further their development. However, little is known about the microbiology of the pouch and how it changes throughout a marsupial’s reproductive cycle. Here, using stringent controls, we characterized the microbial composition of multiple body sites from 26 wild Southern Hairy-nosed Wombats (SHNWs), including pouch samples from animals at different reproductive stages. Results Using qPCR of the 16S rRNA gene we detected a microbial community in the SHNW pouch. We observed significant differences in microbial composition and diversity between the body sites tested, as well as between pouch samples from different reproductive stages. The pouches of reproductively active females had drastically lower microbial diversity (mean ASV richness 19 ± 8) compared to reproductively inactive females (mean ASV richness 941 ± 393) and were dominated by gram positive bacteria from the Actinobacteriota phylum (81.7-90.6%), with the dominant families classified as Brevibacteriaceae, Corynebacteriaceae, Microbacteriaceae, and Dietziaceae. Three of the five most abundant sequences identified in reproductively active pouches had closest matches to microbes previously isolated from tammar wallaby pouches. Conclusions This study represents the first contamination-controlled investigation into the marsupial pouch microbiota, and sets a rigorous framework for future pouch microbiota studies. Our results indicate that SHNW pouches contain communities of microorganisms that are substantially altered by the host reproductive cycle. We recommend further investigation into the roles that pouch microorganisms may play in marsupial reproductive health and joey survival.

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Alkalo-Thermophilic Microbial Diversity of the Tecozautla Geyser, México

10.21203/rs.3.rs-548312/v1 ◽

2021 ◽

Author(s):

Jesus Alberto Segovia-Cruz ◽

Valeria Souza ◽

Yuridia Mercado-Flores ◽

Miguel Angel Anducho-Reyes ◽

Genaro Vargas-Hernández ◽

...

Keyword(s):

16S Rrna ◽

Microbial Communities ◽

Software Package ◽

Microbial Mats ◽

Rrna Gene ◽

Thermal Systems ◽

Microbial Composition ◽

Salt Deposits ◽

The 16S Rrna Gene ◽

Storage Pools

Abstract Microbial mats have been studied in many thermal systems; the most iconic is Yellowstone. In Mexico, the information on microbial mats is scarce and therefore novel. In this research, the thermophilic microbial composition of samples from four areas of the Tecozautla geyser, Hidalgo, Mexico, was studied: sediments (GD), salt deposits (GA), and microbial mats (GB and GC). The samples were taken at the outlet of the geyser (94 °C) and in storage pools with temperatures of 61.5-65 °C. Sequencing of the 16S rRNA gene amplicons was carried out, obtaining 1,425,506 readings, and was analyzed through the Quantitative Insights Into Microbial Ecology software package version 2 (qiime2). 32 phyla were identified in the four samples being the most representative for the GA sample: Armatimonadetes, Chloroflexi, Cyanobacteria, and Thermi, with abundances of 46.35, 19.18, 3.27, and 1.82 %, respectively. For the GB sample, they were Proteobacteria, Bacteroidetes, Cyanobacteria, Spirochaetes, Thermi, and Firmicutes with abundances of 25.23, 22.04, 20.42, 12.31, 4.56, and 1.32 %, respectively. For the GC sample, abundances of 55.60, 9.85, 7.04, 7.01, and 6.15 % were observed for the phylum Chloroflexi, Armatimonadetes, Proteobacteria, Cyanobacteria, and Acidobacteria, respectively. Finally, for the GD sample, the most abundant phyla were Chloroflexi (36.10 %), Cyanobacteria (17.13 %), Armatimonadetes (15.59 %), Proteobacteria (5.45 %), and Nitrospirae with (3.21 %). The metabolic functionality of the microbial communities present in the samples was inferred using the 16S rRNA amplicons. This work represents the first report of the microbial communities present in the Tecozautla geyser.

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Difference of colorectal cancer microbial community by metagenomics and culture-based methods.

Journal of Clinical Oncology ◽

10.1200/jco.2019.37.4_suppl.489 ◽

2019 ◽

Vol 37 (4_suppl) ◽

pp. 489-489

Author(s):

Dong Ho Lee ◽

Mijin Seol ◽

Yu Ra Lee ◽

Young Soo Park ◽

Cheol Min Shin ◽

...

Keyword(s):

Colorectal Cancer ◽

16S Rrna ◽

16S Rrna Gene ◽

Intestinal Microbiota ◽

Agar Plate ◽

Control Group ◽

Rrna Gene ◽

Microbial Composition ◽

Metagenome Analysis ◽

The 16S Rrna Gene

489 Background: Dysbiosis of intestinal microbiota is promoting the development of colorectal cancer (CRC). We confirmed the intestinal microbiota composition from fecal sample of Korean CRC patients. Metagenomic analysis was performed and we isolated single microbes through culture-based method. Methods: CRC fecal samples were collected from 12 individuals. Metagenome Sequencing was based on the 16S rRNA gene amplicon on the Illumina MiSeq platform. The bacteria strains were subcultivated on the agar plate medium in aerobic and anaerobic and further identified by using the 16s rRNA gene sequencing. Results: Bacteria diversity by metagenome analysis was decreased in CRC group compared to control group. In CRC group, relative abundance of Firmicutes and Bacteroidetes were increased while the prevalence of Proteobacteria was decreased. The difference of microbial composition between control and CRC group was found at the genus level. Bacteroides, Parabacteroides of Bacteroidetes have increased and Acinetobacter, Pseudomonas of Proteobacteria have significantly decreased in CRC compared to control group. Using culture method, we isolated diverse bacteria of species level including five strains of Bacteroides; B. ovatus, B. uniformis, B. salyersiae, B. vulgatus and B. xylanisolvens and two strains of Fusobacterium; F. gonidiaformans and F. necrophorum from CRC patients. Conclusions: Metagenome analysis showed the genus Bacteroides, Parabacteroides of the phylum Bacteroidetes has increased and the genus Acinetobacter, Pseudomonas of Proteobacteria decreased in CRC group compared to control group. In addition to, we have isolated various strains associated with CRC by culture-based method.

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First Report of Pantoea agglomerans Causing Brown Apical Necrosis of Walnut in China

Plant Disease ◽

10.1094/pdis-01-11-0060 ◽

2011 ◽

Vol 95 (6) ◽

pp. 773-773 ◽

Cited By ~ 9

Author(s):

K. Q. Yang ◽

W. W. Qu ◽

X. Liu ◽

H. X. Liu ◽

L. Q. Hou

Keyword(s):

16S Rrna ◽

16S Rrna Gene ◽

Juglans Regia ◽

Shandong Province ◽

Pantoea Agglomerans ◽

The Body ◽

Rrna Gene ◽

First Report ◽

Apical Necrosis ◽

The 16S Rrna Gene

Brown apical necrosis (BAN) of walnut (Juglans regia L.) causes premature fruit drop and yield losses and has been reported to be an important walnut production problem in Spain, Italy, France, and Turkey (1,2). A number of organisms have been associated with BAN on walnut: Xanthomonas arboricola pv. juglandis, Fusarium spp., and Alternaria spp. (3). Since the spring of 2007, BAN was observed in 50 to 60% of the trees in walnut orchards in Taian City and Laiwu City, Shandong Province, China. Surface-disinfested tissue from premature walnut fruits was placed onto potato dextrose agar. Alternaria spp., X. arboricola pv. juglandis, and Pantoea agglomerans (formerly Enterobacter agglomerans) were isolated 76, 35, and 45% of the time, respectively. The P. agglomerans cultures formed a yellow lawn and were rod shaped with the body length of 1.5 to 3.0 μm, width of 0.5 to 1.0 μm, and four to six flagella. In biochemical tests, these bacteria were gram negative, lactose positive, and indole negative. Genomic DNA was extracted from one HXJ isolate and the 16S rRNA gene sequence (GenBank Accession No. HM016799) was obtained using universal primers 27F and 1492R. HM016799 had 99% sequence identity with P. agglomerans accessions in GenBank (GU477762, GQ494018, FJ756355, and AB004757). To confirm pathogenicity, HXJ isolate (108 CFU·ml–1) was inoculated at the bottom of the stigma within 5 days after florescence (DAF) and in premature fruit wounded with a needle within 30 DAF in 2008 to 2010. Stigmas injected with only sterile water served as controls. The bacteria were inoculated into three replicate 9-year-old plants of the walnut cv. Xiangling. Forty nuts on each plant were inoculated. The plants were grown in Shandong Province, China (36°09′59″N, 117°13′30″E). Ten days after inoculation, typical internal BAN symptoms were observed on all treated nuts and the controls were still healthy. In the inoculated stigmas, necrosis of stigma and style spread to internal tissues and reached the kernel. In treated premature fruit, internal tissues became necrotic and blackish and eventually led to nut drop. The same bacterium was reisolated from the inoculated tissue. On the basis of morphological, physiological, and biochemical characteristics and sequencing of the 16S rRNA gene, the bacterium was identified as P. agglomerans. To our knowledge, this is the first report of P. agglomerans causing internal type BAN of walnut in China or worldwide. References: (1) A. Belisario et al. Plant Dis. 6:599, 2002. (2) G. Bouvet. Acta Hortic. 705:447, 2005. (3) C. Moragrega and H. Özaktan. J. Plant Pathol. 92:S1.67, 2010.

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Phyllobacterium loti sp. nov. isolated from nodules of Lotus corniculatus

INTERNATIONAL JOURNAL OF SYSTEMATIC AND EVOLUTIONARY MICROBIOLOGY ◽

10.1099/ijs.0.052993-0 ◽

2014 ◽

Vol 64 (Pt_3) ◽

pp. 781-786 ◽

Cited By ~ 33

Author(s):

Maximo Sánchez ◽

Martha-Helena Ramírez-Bahena ◽

Alvaro Peix ◽

María J. Lorite ◽

Juan Sanjuán ◽

...

Keyword(s):

16S Rrna ◽

16S Rrna Gene ◽

Sequence Similarity ◽

Carbon Sources ◽

Lotus Corniculatus ◽

Culture Conditions ◽

Rrna Gene ◽

Content Type ◽

Link Type ◽

The 16S Rrna Gene

Strain S658T was isolated from a Lotus corniculatus nodule in a soil sample obtained in Uruguay. Phylogenetic analysis of the 16S rRNA gene and atpD gene showed that this strain clustered within the genus Phyllobacterium . The closest related species was, in both cases, Phyllobacterium trifolii PETP02T with 99.8 % sequence similarity in the 16S rRNA gene and 96.1 % in the atpD gene. The 16S rRNA gene contains an insert at the beginning of the sequence that has no similarities with other inserts present in the same gene in described rhizobial species. Ubiquinone Q-10 was the only quinone detected. Strain S658T differed from its closest relatives through its growth in diverse culture conditions and in the assimilation of several carbon sources. It was not able to reproduce nodules in Lotus corniculatus. The results of DNA–DNA hybridization, phenotypic tests and fatty acid analyses confirmed that this strain should be classified as a representative of a novel species of the genus Phyllobacterium , for which the name Phyllobacterium loti sp. nov. is proposed. The type strain is S658T( = LMG 27289T = CECT 8230T).

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