scholarly journals Metaproteomic Profiling of Fungal Gut Colonization in Gnotobiotic Mice

2020 ◽  
Author(s):  
Veronika Kuchařová Pettersen ◽  
Antoine Dufour ◽  
Marie-Claire Arrieta
Keyword(s):  
Allergic Airway Inflammation ◽  
Fungal Species ◽  
Fungal Colonization ◽  
Mammalian Host ◽  
Functional Changes ◽  
Disease States ◽  
Gut Colonization ◽  
Increased Sensitivity ◽  
Gnotobiotic Mice ◽  
Eukaryotic Microbes

ABSTRACTEukaryotic microbes can modulate mammalian host health and disease states, yet the molecular contribution of gut fungi remains nascent. We previously showed that mice exclusively colonized with fungi displayed increased sensitivity to allergic airway inflammation and fecal metabolite profiles similar to germ-free mice. To gain insights into the functional changes attributed to fungal colonization, we performed quantitative proteomic analyses of feces and small intestine of four-week-old gnotobiotic mice colonized with bacteria, fungi, or both. A comparison of fecal metaproteomic profiles between the mouse groups yielded broad changes in the relative levels of bacterial (46% of 2,860) and mouse (76% from 405) proteins. Many of the detected fungal proteins (3% of 1,492) have been previously reported as part of extracellular vesicles and having immunomodulating properties. Changes in the levels of mouse proteins derived from the jejunum (4% of 1,514) were mainly driven by proteins functional in lipid metabolism and apoptosis. Using metaproteomic profiling of gnotobiotic conditions, we show that fungal colonization profoundly impacts the host gut proteome. Our results suggest that an increased abundance of certain gut fungal species in early life may impact the developing intracellular balance of epithelial and immune cells.

scholarly journals Metaproteomic Profiling of Fungal Gut Colonization in Gnotobiotic Mice

2021 ◽  
Author(s):  
Veronika Kuchařová Pettersen ◽  
Antoine Dufour ◽  
Marie-Claire Arrieta
Keyword(s):  
Bacterial Species ◽  
Fungal Species ◽  
Mammalian Host ◽  
Marginal Effect ◽  
Host Immune System ◽  
Proteomic Profiling ◽  
Functional Changes ◽  
Fungal Colonisation ◽  
Gut Colonization ◽  
Gnotobiotic Mice

Abstract Background: Eukaryotic microbes can modulate mammalian host health and disease states, yet the molecular contribution of gut fungi remains nascent. We previously showed that mice exclusively colonised with fungi displayed increased sensitivity to allergic airway inflammation and had fecal metabolite profiles similar to germ-free mice. This marginal effect on the host metabolome suggested that fungi do not primarily use metabolites to modulate the host immune system.Methods: To describe functional changes attributed to fungal colonisation, we performed mass spectrometry-based analyses of feces (Label-Free Quantitative; LFQ) and the small intestine (labeling with Tandem Mass Tag; TMT) of gnotobiotic mice colonised with defined consortia of twelve bacterial species, five fungal species, or both. We also evaluated the effect of microbiome perturbances on the metaproteome by analysing feces from mouse pups treated with an antibiotic or antifungal.Results: We detected 6,675 proteins in the mice feces, of which 3,845 had determined LFQ levels. Analysis of variance showed changes in the different gnotobiotic mouse groups; specifically, 46% of 2,860 bacterial, 15% of 580 fungal, and 76% of 405 mouse quantified proteins displayed differential levels. The antimicrobial treatments resulted in lasting changes in the bacterial and fungal proteomes, suggesting that the antimicrobials impacted the entire community. Fungal colonisation resulted in changes in host proteins functional in innate immunity as well as metabolism, predicting specific roles of gut fungi on host systems during early developmental stages. Several of the detected fungal proteins (3% of 1,492) have been previously reported as part of extracellular vesicles and having immunomodulating properties. Using an isobaric labelling TMT approach for profiling low abundant proteins of the jejunal tissue, we confirmed that the five fungal species differentially impacted the host intestinal proteome compared to the bacterial consortium. The detected changes in mouse jejunal proteins (4% of 1,514) were mainly driven by metabolic proteins. Conclusions: We used quantitative proteomic profiling of gnotobiotic conditions to show how colonisation with selected fungal species impacts the host gut proteome. Our results suggest that an increased abundance of certain gut fungal species in early life may affect the developing intracellular attributes of epithelial and immune cells.

scholarly journals Molds in museum environments: Biodeterioration of art photographs and wooden sculptures

2013 ◽  
Vol 65 (3) ◽  
pp. 955-962 ◽  
Author(s):  
Milica Ljaljevic-Grbic ◽  
M. Stupar ◽  
Jelena Vukojevic ◽  
Ivana Maricic ◽  
Natasa Bungur
Keyword(s):  
Aspergillus Niger ◽  
Indoor Air ◽  
Trichoderma Viride ◽  
Fungal Species ◽  
Chaetomium Globosum ◽  
Fungal Colonization ◽  
Cellulolytic Activity ◽  
Cultural Center ◽  
Fungal Propagules ◽  
Alternaria Sp

Pieces of art stored in museum depots and display rooms are subject to fungal colonization that leads to bio-deterioration processes. Deteriorated wooden sculptures and art photographs temporarily stored in the quarantine room of the Cultural Center of Belgrade were subject to mycological analyses. Twelve fungal species were identified on the wooden substratum and five species were detected on photograph surfaces. Trichoderma viride, Chaetomium globosum and Alternaria sp. were the fungi with proven cellulolytic activity detected on the examined cellulose substrata. Indoor air mycobiota were estimated to 210.09 ? 8.06 CFU m-3, and the conidia of fungus Aspergillus niger were the dominant fungal propagules in the air of the examined room.

scholarly journals Pre- and Neonatal Imprinting on Immunological Homeostasis and Epithelial Barrier Integrity by Escherichia coli Nissle 1917 Prevents Allergic Poly-Sensitization in Mice

2021 ◽  
Vol 11 ◽  
Author(s):  
Priya J. Sarate ◽  
Dagmar Srutkova ◽  
Nora Geissler ◽  
Martin Schwarzer ◽  
Irma Schabussova ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Grass Pollen ◽  
Allergic Airway Inflammation ◽  
Experimental Models ◽  
Lung Epithelial Cells ◽  
Probiotic Bacteria ◽  
Allergy Prevention ◽  
Lactation Period ◽  
Gnotobiotic Mice ◽  
Immunological Homeostasis

A steady rise in the number of poly-sensitized patients has increased the demand for effective prophylactic strategies against multi-sensitivities. Probiotic bacteria have been successfully used in clinics and experimental models to prevent allergic mono-sensitization. In the present study, we have investigated whether probiotic bacteria could prevent poly-sensitization by imprinting on the immune system early in life. We used two recombinant variants of probiotic Escherichia coli Nissle 1917 (EcN): i) EcN expressing birch and grass pollen, poly-allergen chimera construct (EcN-Chim), and ii) an “empty” EcN without allergen expression (EcN-Ctrl). Conventional mice (CV) were treated with either EcN-Chim or EcN-Ctrl in the last week of the gestation and lactation period. Gnotobiotic mice received one oral dose of either EcN-Chim or EcN-Ctrl before mating. The offspring from both models underwent systemic allergic poly-sensitization and intranasal challenge with recombinant birch and grass pollen allergens (rBet v 1, rPhl p 1, and rPhl p 5). In the CV setting, the colonization of offspring via treatment of mothers reduced allergic airway inflammation (AAI) in offspring compared to poly-sensitized controls. Similarly, in a gnotobiotic model, AAI was reduced in EcN-Chim and EcN-Ctrl mono-colonized offspring. However, allergy prevention was more pronounced in the EcN-Ctrl mono-colonized offspring as compared to EcN-Chim. Mono-colonization with EcN-Ctrl was associated with a shift toward mixed Th1/Treg immune responses, increased expression of TLR2 and TLR4 in the lung, and maintained levels of zonulin-1 in lung epithelial cells as compared to GF poly-sensitized and EcN-Chim mono-colonized mice. This study is the first one to establish the model of allergic poly-sensitization in gnotobiotic mice. Using two different settings, gnotobiotic and conventional mice, we demonstrated that an early life intervention with the EcN without expressing an allergen is a powerful strategy to prevent poly-sensitization later in life.

scholarly journals Wild Apple-Associated Fungi and Bacteria Compete to Colonize the Larval Gut of an Invasive Wood-Borer Agrilus mali in Tianshan Forests

2021 ◽  
Vol 12 ◽  
Author(s):  
Tohir A. Bozorov ◽  
Zokir O. Toshmatov ◽  
Gulnaz Kahar ◽  
Daoyuan Zhang ◽  
Hua Shao ◽  
...  
Keyword(s):  
Gut Microbiota ◽  
Inhibitory Concentration ◽  
Fungal Species ◽  
Gut Bacteria ◽  
Recent Analysis ◽  
Fungal Colonization ◽  
Antifungal Compound ◽  
Concentration Method ◽  
Malus Sieversii ◽  
Wood Borer

The gut microflora of insects plays important roles throughout their lives. Different foods and geographic locations change gut bacterial communities. The invasive wood-borer Agrilus mali causes extensive mortality of wild apple, Malus sieversii, which is considered a progenitor of all cultivated apples, in Tianshan forests. Recent analysis showed that the gut microbiota of larvae collected from Tianshan forests showed rich bacterial diversity but the absence of fungal species. In this study, we explored the antagonistic ability of the gut bacteria to address this absence of fungi in the larval gut. The results demonstrated that the gut bacteria were able to selectively inhibit wild apple tree-associated fungi. Among them, Pseudomonas synxantha showed strong antagonistic ability, producing antifungal compounds. Using different analytical methods, such as column chromatography, mass spectrometry, HPLC, and NMR, an antifungal compound, phenazine-1-carboxylic acid (PCA), was identified. Activity of the compound was determined by the minimum inhibitory concentration method and electron microscopy. Moreover, our study showed that the gut bacteria could originate from noninfested apple microflora during infestation. Overall, the results showed that in newly invaded locations, A. mali larvae changed their gut microbiota and adopted new gut bacteria that prevented fungal colonization in the gut.

Chronic exposure to ozone causes tolerance to airway hyperresponsiveness in guinea pigs: lack of SOD role

1998 ◽  
Vol 84 (5) ◽  
pp. 1749-1755 ◽  
Author(s):  
Mario H. Vargas ◽  
Laura Romero ◽  
Bettina Sommer ◽  
Pedro Zamudio ◽  
Pascal Gustin ◽  
...  
Keyword(s):  
Guinea Pigs ◽  
Airway Hyperresponsiveness ◽  
Chronic Exposure ◽  
Neutral Endopeptidase ◽  
Respiratory Effects ◽  
Functional Changes ◽  
Independent Mechanism ◽  
Increased Sensitivity ◽  
Pressure Changes

Tolerance to respiratory effects of O3 has been demonstrated for anatomic and functional changes, but information about tolerance to O3-induced airway hyperresponsiveness (AHR) is scarce. In guinea pigs exposed to air or O3 (0.3 parts/million, 4 h/day, for 1, 3, 6, 12, 24, or 48 days, studied 16–18 h later), pulmonary insufflation pressure changes induced by intravenous substance P (SP, 0.032–3.2 μg/kg) were measured, then the animals were subjected to bronchoalveolar lavage (BAL). Bronchial rings with or without phosphoramidon were also evaluated 3 h after air or a single O3 exposure. O3 caused in vivo AHR (increased sensitivity) to SP after 1, 3, 6, 12, and 24 days of exposure compared with control. However, after 48 days of exposure, O3 no longer caused AHR. Total cell, macrophage, neutrophil, and eosinophil counts in BAL were increased in most O3-exposed groups. When data from all animals were pooled, we found a highly significant correlation between degree of airway responsiveness and total cells ( r = 0.55), macrophages ( r = 0.54), neutrophils ( r = 0.47), and eosinophils ( r = 0.53), suggesting that airway inflammation is involved in development of AHR to SP. Superoxide dismutase (SOD) levels in BAL fluids were increased ( P < 0.05) after 1, 3, 6, and 12 days of O3 exposure and returned to basal levels after 24 and 48 days of exposure. O3 failed to induce hyperresponsiveness to SP in bronchial rings, and phosphoramidon increased responses to SP in air- and O3-exposed groups, suggesting that neutral endopeptidase inactivation was not involved in O3-induced AHR to SP in vivo. We conclude that chronic exposure to 0.3 ppm O3, a concentration found in highly polluted cities, resulted in tolerance to AHR to SP in guinea pigs by an SOD-independent mechanism.

scholarly journals Cohort Study of Airway Mycobiome in Adult Cystic Fibrosis Patients: Differences in Community Structure between Fungi and Bacteria Reveal Predominance of Transient Fungal Elements

2015 ◽  
Vol 53 (9) ◽  
pp. 2900-2907 ◽  
Author(s):  
Rolf Kramer ◽  
Annette Sauer-Heilborn ◽  
Tobias Welte ◽  
Carlos A. Guzman ◽  
Wolf-Rainer Abraham ◽  
...  
Keyword(s):  
Cystic Fibrosis ◽  
Fungal Spores ◽  
Rare Event ◽  
Single Strand Conformation Polymorphism ◽  
Fungal Species ◽  
Fungal Communities ◽  
Fungal Colonization ◽  
Molecular Fingerprinting ◽  
High Exposure ◽  
Entire Cohort

The respiratory mycobiome is an important but understudied component of the human microbiota. Like bacteria, fungi can cause severe lung diseases, but their infection rates are much lower. This study compared the bacterial and fungal communities of sputum samples from a large cohort of 56 adult patients with cystic fibrosis (CF) during nonexacerbation periods and under continuous antibiotic treatment. Molecular fingerprinting based on single-strand conformation polymorphism (SSCP) analysis revealed fundamental differences between bacterial and fungal communities. Both groups of microorganisms were taxonomically classified by identification of gene sequences (16S rRNA and internal transcript spacer), and prevalences of single taxa were determined for the entire cohort. Major bacterial pathogens were frequently observed, whereas fungi of known pathogenicity in CF were detected only in low numbers. Fungal species richness increased without reaching a constant level (saturation), whereas bacterial richness showed saturation after 50 patients were analyzed. In contrast to bacteria, a large number of fungal species were observed together with high fluctuations over time and among patients. These findings demonstrated that the mycobiome was dominated by transient species, which strongly suggested that the main driving force was their presence in inhaled air rather than colonization. Considering the high exposure of human airways to fungal spores, we concluded that fungi have low colonization abilities in CF, and colonization by pathogenic fungal species may be considered a rare event. A comprehensive understanding of the conditions promoting fungal colonization may offer the opportunity to prevent colonization and substantially reduce or even eliminate fungus-related disease progression in CF.

scholarly journals Constitutive Delivery of Bovine β-Lactoglobulin to the Digestive Tracts of Gnotobiotic Mice by Engineered Lactobacillus casei

2006 ◽  
Vol 72 (12) ◽  
pp. 7460-7467 ◽  
Author(s):  
S. Hazebrouck ◽  
R. Oozeer ◽  
K. Adel-Patient ◽  
P. Langella ◽  
S. Rabot ◽  
...  
Keyword(s):  
Immune System ◽  
Lactobacillus Casei ◽  
Allergic Sensitization ◽  
Total Production ◽  
Postnatal Maturation ◽  
Gut Colonization ◽  
Gnotobiotic Mice ◽  
Allergic Disorders ◽  
Β Lactoglobulin

ABSTRACT The gut microbiota is critical for maturation of the immune system. Recent evidence suggests that early establishment of lactobacilli in the intestinal microbiota, during neonatal colonization or by probiotic supplementation, could prevent the development of allergic disorders. Postnatal maturation of the gut immune system with allergen-producing lactobacilli colonizing the digestive tract could then affect the development of further allergic sensitization. In this paper, we describe construction of a recombinant Lactobacillus casei strain that can constitutively deliver bovine β-lactoglobulin (BLG), a major cow's milk allergen, to the guts of gnotobiotic mice. The blg gene was inserted into the L. casei chromosome downstream of an endogenous promoter. BLG production was improved by fusing the propeptide LEISSTCDA (LEISS) to the BLG mature moiety. This led to a 10-fold increase in LEISS-BLG production compared to the production obtained without the propeptide and also led to enhanced secretion corresponding to 5% of the total production. After inoculation into germfree C3H/HeN mice, the genetic stability of the recombinant strain and in vivo BLG production were confirmed for at least 10 weeks. BLG stimulation of spleen cells from mice monoassociated with the BLG-producing lactobacilli induced secretion of the Th1 cytokine gamma interferon and, to a lesser extent, the Th2 cytokine interleukin-5. No BLG-specific immunoglobulin G1 (IgG1), IgG2a, or IgA was detected in sera or in fecal samples. These results suggest that gut colonization with allergen-producing lactobacilli could provide a useful model for studying the modulation of allergic disorders.

scholarly journals Mutual Exclusion between Fungal Species of the Fusarium Head Blight Complex in a Wheat Spike

2015 ◽  
Vol 81 (14) ◽  
pp. 4682-4689 ◽  
Author(s):  
Dorothée Siou ◽  
Sandrine Gélisse ◽  
Valérie Laval ◽  
Frédéric Suffert ◽  
Christian Lannou
Keyword(s):  
Mutual Exclusion ◽  
Field Studies ◽  
Fungal Species ◽  
Microdochium Nivale ◽  
Fungal Colonization ◽  
Head Blight ◽  
Classical Analysis ◽  
Content Type ◽  
Single Spike ◽  
Wheat Spike

ABSTRACTFusariumhead blight (FHB) is one of the most damaging diseases of wheat. FHB is caused by a species complex that includes two genera of Ascomycetes:MicrodochiumandFusarium. Fusarium graminearum,Fusariumculmorum,Fusariumpoae, andMicrodochium nivaleare among the most common FHB species in Europe and were chosen for these experiments. Field studies and surveys show that two or more species often coexist within the same field or grain sample. In this study, we investigated the competitiveness of isolates of different species against isolates ofF. graminearumat the scale of a single spike. By performing point inoculations of a single floret, we ensured that each species was able to establish independent infections and competed for spike colonization only. The fungal colonization was assessed in each spike by quantitative PCR. After establishing that the spike colonization was mainly downwards, we compared the relative colonization of each species in coinoculations. Classical analysis of variance suggested a competitive interaction but remained partly inconclusive because of a large between-spike variance. Further data exploration revealed a clear exclusion of one of the competing species and the complete absence of coexistence at the spike level.

scholarly journals Evaluating the Reaction of Peach Cultivars to Infection by Three Botryosphaeria Species

HortScience ◽  
1990 ◽  
Vol 25 (4) ◽  
pp. 468-470 ◽  
Author(s):  
K.O. Britton ◽  
F.F. Hendrix ◽  
P.L. Pusey ◽  
W.R. Okie ◽  
C.C. Reilly ◽  
...  
Keyword(s):  
Growth Rate ◽  
Disease Severity ◽  
Tree Growth ◽  
Field Experiments ◽  
Fungal Species ◽  
Fungal Colonization ◽  
Peach Tree ◽  
Greenhouse Study ◽  
Vascular Discoloration ◽  
Cultivar Susceptibility

Two field experiments were conducted to assess peach (Prurus persica L.) cultivar susceptibility to the three Botryosphaeria spp. that cause peach tree fungal gummosis. Inoculated trees were evaluated for disease severity by rating gum exudation, vascular discoloration, and fungal colonization. Each severity measurement yielded a different rank ordering of cultivars for susceptibility. However, in a greenhouse study, these same measurements gave consistent rankings for aggressiveness of the fungal species on `Blake'. Despite large differences in disease severity in the greenhouse study, none of the severity measures were correlated with tree growth after inoculation. The only factor significantly correlated with growth rate of the trees after inoculation was growth rate before inoculation.

Fungi associated with Tomicus minor on Pinus sylvestris in Poland and their succession into the sapwood of beetle-infested windblown trees

2008 ◽  
Vol 38 (10) ◽  
pp. 2579-2588 ◽  
Author(s):  
Robert Jankowiak
Keyword(s):  
Species Composition ◽  
Pinus Sylvestris ◽  
Initial Phase ◽  
Bark Beetles ◽  
General Scheme ◽  
Fungal Species ◽  
Fungal Colonization ◽  
Fungal Succession ◽  
Tomicus Minor ◽  
Insect Fungi

The study identified and measured frequency of fungal species associated with Tomicus minor (Hart.) on Pinus sylvestris L. (Scots pine) in Poland. Additionally, fungal succession in P. sylvestris sapwood was investigated during a 12 week period following an attack by this insect. Fungi were isolated from five populations of overwintered adult beetles and their galleries with 59 species of fungi being represented among the 2880 cultures obtained. The most frequent species, Ophiostoma canum (Münch) Syd. & P. Syd., Hormonema dematioides Lagerb. & Melin, and Ambrosiella tingens (Lagerb. & Melin) L.R. Batra, appeared to be specifically associated with T. minor. The succesional changes in species composition during a 12 week period following an attack by T. minor were observed. The pattern of fungal succession in P. sylvestris sapwood essentially agreed with a general scheme of fungal succession in tree sapwood infested by bark beetles. Ambrosiella tingens was the first invader of sapwood and occurred most frequently in its deeper layers. Ophiostoma canum, H. dematioides, and other molds were also often isolated from the sapwood; however, they were most common at a depth of 5 mm during the initial phase of fungal colonization. Later, Ophiostoma canum followed A. tingens in the sapwood invasion.

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